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Sample records for culture medium suggesting

  1. New culture medium concepts for cell transplantation.

    Science.gov (United States)

    Lee, S; Kim, B Y; Yeo, J E; Nemeno, J G; Jo, Y H; Yang, W; Nam, B M; Namoto, S; Tanaka, S; Sato, M; Lee, K M; Hwang, H S; Lee, J I

    2013-10-01

    Before cell or tissue transplantation, cells or tissues have to be maintained for a certain period in vitro using culture medium and methods. Most culture media contain substances such as pH indicators and buffers. It is not known whether some of these substances are safe for subsequent application in the transplantation of cells or tissues into the human body. We investigated culture media and methods with respect to the safety of the components in future transplantation applications. A modified culture medium--medical fluid-based culture medium (FCM)--was designed by using various fluids and injectable drugs that are already currently permitted for use in clinical medicine. Medium components necessary for optimal cell growth were obtained from approved drugs. FCM was manufactured with adjusted final concentrations of the medium components similar to those in commercial Dulbecco's modified Eagle's medium (DMEM). In particular, 1029.40 mg/L amino acids, approximately 88.85 mg/L vitamins, 13,525.77 mg/L inorganic salts, and 4500 mg/L D-glucose comprise the high-glucose FCM. Next, human fat synovium-derived mesenchymal stem cells and rat H9c2 (2-1) cells were cultured under 2 conditions: (1) DMEM-high glucose (HG), an original commercial medium, and (2) optimized FCM-HG. We assessed the morphologies and proliferation rates of these cells. We observed that FCM-HG was able to induce the growth of FS-MSC and commercially available H9c2 cell. The morphologies and proliferation patterns of these cells cultured under FCM-HG showed no differences compared with cells grown in DMEM-HG. Our data suggest that FCM, which we developed for the first time according to the concept of drug repositioning, was a useful culture medium, especially in cultured cells intended for human cell transplantation. Crown Copyright © 2013. Published by Elsevier Inc. All rights reserved.

  2. 21 CFR 866.2350 - Microbiological assay culture medium.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Microbiological assay culture medium. 866.2350... Microbiological assay culture medium. (a) Identification. A microbiological assay culture medium is a device that... organism in the innoculated medium. Test results aid in the diagnosis of disease resulting from either...

  3. 21 CFR 866.2390 - Transport culture medium.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Transport culture medium. 866.2390 Section 866... medium. (a) Identification. A transport culture medium is a device that consists of a semisolid, usually non-nutrient, medium that maintains the viability of suspected pathogens contained in patient...

  4. 21 CFR 866.2330 - Enriched culture medium.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Enriched culture medium. 866.2330 Section 866.2330... medium. (a) Identification. An enriched culture medium is a device that consists primarily of liquid or... medium enriched by the addition of such nutritional components as blood, blood serum, vitamins, and...

  5. 21 CFR 866.2320 - Differential culture medium.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Differential culture medium. 866.2320 Section 866... medium. (a) Identification. A differential culture medium is a device that consists primarily of liquid... biochemical component(s) to the medium. Microorganisms are identified by a visible change (e.g., a color...

  6. INNOVATIVE CULTURE IN SMALL AND MEDIUM ENTERPRISES

    Directory of Open Access Journals (Sweden)

    Aluisio Broering Mambrini

    2011-10-01

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

  7. 21 CFR 866.2360 - Selective culture medium.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Selective culture medium. 866.2360 Section 866.2360 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... medium. (a) Identification. A selective culture medium is a device that consists primarily of liquid or...

  8. 21 CFR 866.2300 - Multipurpose culture medium.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Multipurpose culture medium. 866.2300 Section 866.2300 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... medium. (a) Identification. A multipurpose culture medium is a device that consists primarily of liquid...

  9. Suggestions for English Culture Teaching in High School

    Institute of Scientific and Technical Information of China (English)

    Cai Hongjuan

    2016-01-01

    With the implementation of the new High School English Curriculum Standards, more and more people have realized the importance of English culture teaching. To realize the goals of English teaching, teachers should cultivate students' culture awareness and develop their intercultural communicative competence. But in the actual teaching, culture teaching did not get real implementation. So the author puts forwards some suggestions for English culture teaching in high school.

  10. Micropropagation of orchid hybrids in knudson culture medium with addiction of vitamins of ms culture medium, benzilaminopurine and activated charcoal

    Directory of Open Access Journals (Sweden)

    Fabíola Villa

    2014-02-01

    Full Text Available The genera Cattleya and Brassavola, naturally occurring in Brazil, are widely used in hybridization to obtain better shape of the lip in the hybrids. This demand creates the need to develop more effective propagation to meet the market and contributing to avoid the extinction of these plants. This study aimed to test BAP concentrations, activated charcoal, culture medium Knudson and culture medium MS’s vitamins in subculture of Brassocattleya Pastoral x Laeliocattleya Amber Glow orchids plants. There were used seedlings from in vitro germination, with 1-1.5 cm length, undergoing standardization in the culture medium Knudson for three months. The experiment were performed using concentrations of culture medium Knudson (0%, 50%, 100%, 150% and 200%, combined with concentrations of culture medium MS’s vitamins (0%, 50%, 100% and 200% and BAP concentrations (0; 1.0; 2.0 and 4.0 mg L-1 versus activated charcoal (0, 200, 400 and 600 mg L-1, incorporated in a culture medium Knudson. In each flask of 250 mL containing approximately 50 mL of culture medium, four seedlings were placed under aseptic conditions. It was concluded that can use 129% of Knudson, supplemented with 137.5 mg L-1 activated charcoal, 104% of vitamin MS medium without addition of BAP as the best composition for culture medium of hybrid orchid in vitro propagation. The Knudson culture medium does not promote increase in the root system.

  11. Development of a selective culture medium for Fusarium moniliforme.

    Science.gov (United States)

    Castellá, G; Bragulat, M R; Rubiales, M V; Cabañes, F J

    1997-12-01

    Nash and Snyder medium and malachite green agar 2.5 ppm medium, a new selective culture medium designed in our laboratory, were challenged with pure cultures of Fusarium moniliforme strains and two different mixed-conidium suspensions, which included rapidly spreading fungi, for their utility in the isolation and enumeration of F. moniliforme. From the results of this comparative study, malachite green agar 2.5 ppm allowed only the selective growth of F. moniliforme whereas Nash and Snyder medium allowed both the growth of F. moniliforme and other species not belonging to Fusarium spp. The enumeration of F. moniliforme propagules was similar in both culture media.

  12. Isolated Cells of Porphyra yezoensis Cultured on Solid Medium

    Institute of Scientific and Technical Information of China (English)

    沈颂东; 戴继勋

    2001-01-01

    Vegetative cells of Porphyra yezoensis are isolated with sea snail enzyme and cultured on the solidified agar medium. The results of experiments show that the isolated cells can survive,divide and regenerate well on the medium solidified with agar. The first division on the solid medium starts after 7 days' culture, 4 days later than the liquid culture. The survival rate of isolated cells is 71.3% on the solid medium, lower than the 86.2% of that in seawater.Thalli, thalloids,conchocelis, spermatangia and multicellular masses are developed on the solid/medium in the first month, slowly but normally. Spermatangia sacs disappear within 4 weeks. Without adding nutrient liquid onto the surface of solid medium or injecting seawater under the agar layer in order to keep moisture, the thalli and cell groups release monospores to form new thalli instead of enlarging their areas after 5 weeks' culturing. Some monospores regenerate new thalli. Other monospores lose their pigments and minimize their volume and divide quickly to form light pink calli. After 16 weeks, numerous calli can be seen on the solid medium and after 24 weeks' culturing, almost only calli and conchocelis can be seen. If the calli are immersed in seawater, the monospores are released and may develop into young thallus.

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

    NARCIS (Netherlands)

    Robinson, J.W.

    2004-01-01

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

  14. Electromechanical and Elastic Probing of Bacteria in Cell Culture Medium

    Science.gov (United States)

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

    2012-01-01

    Rapid phenotype characterization and identification of cultured cells, which is needed for progress in tissue engineering and drug testing, requires an experimental technique that measures physical properties of cells with sub-micron resolution. Recently, band excitation piezoresponse force microscopy (BEPFM) has been proven useful for recognition and imaging of different types of bacteria in pure water. Here, the BEPFM method is performed for the first time in physiologically-relevant electrolyte media, such as Dulbecco’s phosphate-buffered saline (DPBS) and Dulbecco’s modified Eagle’s medium (DMEM). Distinct electromechanical responses for Micrococcus lysodeikticus (Gram-positive) and Pseudomonas fluorescens (Gram-negative) bacteria are demonstrated in DPBS. The results suggest that mechanical properties of the outer surface coating each bacterium, as well as the electrical double layer around them, are responsible for the BEPFM image formation mechanism in electrolyte media. PMID:22641388

  15. Developing hazelnut tissue culture medium free of ion confounding

    Science.gov (United States)

    The general approach for tissue culture medium optimization is to use salts as factors in experimental design and analysis. However, using salts as factors leads to ion confounding, making it difficult to detect the effects of individual ions on particular growth responses. This study focused on tes...

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

    African Journals Online (AJOL)

    ajl yemi

    2011-11-11

    Nov 11, 2011 ... its ability to form aqueous gels, dispersion stabilizer. There are two main ... Protopectinases (PPases) are used as heterogeneous group of ... one day, and then maintained at 4°C. The seed culture medium contained (g/L): ...

  17. Culture medium for amylase production by toxigenic fungi

    Directory of Open Access Journals (Sweden)

    Figueira Edson Luiz Zangrando

    2000-01-01

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

  18. [Evaluation of chromogenic medium Uriselect4 in urine culture].

    Science.gov (United States)

    Ferjani, Asma; Marzouk, Manel; Idriss, Nadia; Sammoud, Sammoud; Hannachi, Naila; Boukadida, Janel

    2011-01-01

    We evaluated the performance and the cost of chromogenic medium Uriselect4 agar with regard to the standard medium for the detection and identification of urinary tract pathogens. A total of 503 clinical urine specimens containing leucocytes greater or equal to 104/mL were analysed prospectively, in parallel by two different persons on blood agar (GS) and Uriselect4 according to the manufacturers' instructions. Of the 503 urine specimens tested, 210 gave a positive culture on Uriselect4 versus 181 on GS. The majority of bacterial species grew on both media; enterobacteria grew on Uriselect4 better than GS. The identification of Escherichia coli (E. coli), Proteus mirabilis (P. mirabilis), KES group and Enterococcus faecalis (E. faecalis) did not require the use of galleries Api and has a gain of 24  h. Positive pure cultures on Uriselect4 corresponding to negative cultures of GS were noted in 17 ases. Conversely, in seven cases a positive pure culture on GS was noted while the corresponding Uriselect4 cultures were negative. The cost of identification on GS (including the cost of galleries Api), was about two times higher than Uriselect4. Uriselect4 medium isolates the most frequent urinary tract pathogens and identify them so almost immediately, with a lower cost.

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

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Culture medium for antimicrobial susceptibility....1700 Culture medium for antimicrobial susceptibility tests. (a) Identification. A culture medium for antimicrobial susceptibility tests is a device intended for medical purposes that consists of any medium capable...

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

    Science.gov (United States)

    Mahoney, Mark M.; Staley, Kevin J.

    2017-01-01

    Rodent organotypic hippocampal cultures spontaneously develop epileptiform activity after approximately 2 weeks in vitro and are increasingly used as a model of chronic post-traumatic epilepsy. However, organotypic cultures are maintained in an artificial environment (culture medium), which contains electrolytes, glucose, amino acids and other components that are not present at the same concentrations in cerebrospinal fluid (CSF). Therefore, it is possible that epileptogenesis in organotypic cultures is driven by these components. We examined the influence of medium composition on epileptogenesis. Epileptogenesis was evaluated by measurements of lactate and lactate dehydrogenase (LDH) levels (biomarkers of ictal activity and cell death, respectively) in spent culture media, immunohistochemistry and automated 3-D cell counts, and extracellular recordings from CA3 regions. Changes in culture medium components moderately influenced lactate and LDH levels as well as electrographic seizure burden and cell death. However, epileptogenesis occurred in any culture medium that was capable of supporting neural survival. We conclude that medium composition is unlikely to be the cause of epileptogenesis in the organotypic hippocampal culture model of chronic post-traumatic epilepsy. PMID:28225808

  1. Comparative evaluation of cytotoxicity of cadmium in rat liver cells cultured in serum-containing medium and commercially available serum-free medium.

    Science.gov (United States)

    Latinwo, Lekan M; Badisa, Veera L D; Odewumi, Caroline O; Ikediobi, Christopher O; Badisa, Ramesh B; Brooks-Walter, Alexis; Lambert, Ayuk-Takem T; Nwoga, Jude

    2008-07-01

    Cadmium (Cd) is an industrial pollutant and carcinogenic metal. Most in vitro Cd toxicity studies have been carried out in various cell lines cultured in 10% fetal bovine serum (FBS) containing medium. In this report, we compared the toxic effect of Cd (0-300 microM) on cell growth, total RNA, total proteins, and antioxidant enzymes in rat normal liver cells cultured in medium with 10% FBS or commercially available serum-free medium for 4 or 8 hours. With Cd concentration at above 100 microM, the total levels of RNA, protein and cell growth decreased in serum-containing medium, while their levels increased in serum-free medium compared to the controls. The glutathione peroxidase and glutathione reductase levels were lower in serum-free medium than in serum-containing medium, indicating less oxidative stress in cells grown in serum-free medium. These results clearly suggest that Cd showed higher toxicity to liver cells grown in serum-containing medium in comparison to commercially available serum-free medium. It is speculated that albumin and other substances present in commercial serum-free medium chelated Cd and thereby protected these cells against Cd toxicity. Even under in vivo conditions, cadmium enters into various organs after passing through blood which contains serum. Based on these studies, it appears that media containing serum may be ideal for in vivo toxicity correlation studies with animal cells.

  2. Dense cultures of Neisseria gonorrhoeae in liquid medium.

    Science.gov (United States)

    Brookes, R; Hedén, C G

    1967-03-01

    Cultivation of Neisseria gonorrhoeae was effected in a conical glass culture vessel surrounded by a constant-temperature water jacket, and with facilities for stirring, aeration, and pH measurement and control. With the use of an aerated peptone-based medium, containing polypropylene glycol to prevent foam build-up, the yields obtained over the pH range from 5.8 to 7.4 were determined. The greatest yield was obtained at pH 6.4 when the dry weight was 1.5 g/liter. At pH 7.2 to 7.6, lysis was extensive.

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

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Culture medium for pathogenic Neisseria spp. 866.2410 Section 866.2410 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... Culture medium for pathogenic Neisseria spp. (a) Identification. A culture medium for pathogenic Neisseria...

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

    Science.gov (United States)

    Kumar, Anup; Singh, Neera

    2016-03-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

  6. [Improved culturability of soil bacteria using proper combination with various culturing medium].

    Science.gov (United States)

    Hu, Yuan-Sen; Li, Cui-Xiang; Sun, Fu-Lin; Wu, Kun; Jia, Xin-Cheng

    2007-10-01

    To isolate more unique and previously unrecognized bacteria in soil samples, the culture difference under three incubation modes was investigated by using trophic, low-nutrient broth and soil extract as growth medium. Plate count proved that the oligotrophic medium resulted in a slow growth and consecutive colony formation over the course of incubation. On the 5th day, the most number of colony-forming unit was found on trophic LB and low-nutrient R2A, which was approximate 5 times as many as that isolated on 0.1 x LB. Of the 7 media, LB broth harvested the maximum bacterial communities, and novel species could be isolated as the nutrient was diluted to appropriate extent. The DGGE patterns of oligotrophic and rich nutrient culture collection displayed low similarity, however, the bands at various lanes exhibited complementary effect. When cultivated with static flask, LB and R2A media obtained more bacterial species, which concluded most species isolated by the other five media. Under the test tube incubation mode, the most species was also found in LB medium except some appeared only on R2A and TSB. Apparent bacterial communities difference could be detected between R2A, LB and TSB media. The experiment data may contribute much to the special medium design as well as improvement of bacterial culturability by using proper medium.

  7. Optimizing culture medium for meristem tissue culture of several Saccharum species and commercial hybrids

    Science.gov (United States)

    The optimal range of medium nutrients and plant growth regulators (PGR) was investigated for in vitro culture of diverse sugarcane species and cultivars. Macro-nutrients, nitrogen (N), phosphorous (P) and potassium (K), were essential for growth of leaf primordia. Although the best concentration of ...

  8. Calcification in human osteoblasts cultured in medium conditioned by the prostatic cancer cell line PC-3 and prostatic acid phosphatase.

    Science.gov (United States)

    Kimura, G; Sugisaki, Y; Masugi, Y; Nakazawa, N

    1992-01-01

    A medium that had been conditioned by PC-3 cells stimulated the calcification of a human osteoblastic cell line, Tak-10, in a nonmitogenic culture. The calcification of the osteoblasts was stimulated maximally at a 25% concentration of the conditioned medium. Calcification activity was markedly enhanced by the addition of both prostatic acid phosphatase (PAP) and its substrate, alpha-glycerophosphate, to the medium; however, PAP added alone did not enhance this activity. These results suggest that human prostatic carcinoma cells produce a factor that stimulates the calcification of the human osteoblasts. Results have also suggested that PAP is a requisite for osteogenesis provided that its substrates are abundant in the medium.

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

    Science.gov (United States)

    Suzuki, Tadahiro; Iwahashi, Yumiko

    2016-01-01

    Aflatoxin (AF) is a harmful secondary metabolite that is synthesized by the Aspergillus species. Although AF detection techniques have been developed, techniques for detection of AF synthetic fungi are still required. Techniques such as plate culture methods are continually being modified for this purpose. However, plate culture methods require refinement because they suffer from several issues. In this study, activated charcoal powder (carbon) was added to a culture medium containing cyclodextrin (CD) to enhance the contrast of fluorescence and improve the detection efficiency for AF synthetic fungi. Two culture media, potato dextrose agar and yeast extract sucrose agar, were investigated using both plate and liquid cultures. The final concentrations of CD and carbon in the media were 3 mg/mL and 0.3 mg/mL, respectively. Addition of carbon improved the visibility of fluorescence by attenuating approximately 30% of light scattering. Several fungi that could not be detected with only CD in the medium were detected with carbon addition. The carbon also facilitated fungal growth in the potato dextrose liquid medium. The results suggest that addition of carbon to media can enhance the observation of AF-derived fluorescence. PMID:27854283

  10. Glycosidases from the culture medium of Physarum polycephalum.

    Science.gov (United States)

    Kilpatrick, D C; Stirling, J L

    1977-01-01

    Eight exo-glycosidase activities were detected in the axenic culture medium of the myxomycete, Physarum polycephalum. The secretion of each enzyme examined followed the growth curve and continued during the stationary phase after the cessation of growth. Two or more forms of each enzyme were detected after electrophoretic separation. The beta-N-acetyl-D-hexosaminidase activity was readily separated into its two electrophoretic forms, X and Y, which were purified 145- and 306-fold respectively. These beta-N-acetyl-D-hexosaminidases had several similar characteristics. Evidence is presented that the major electrophoretic form of alpha-D-galactosidase is heterogeneous. The possible functions of extracellular glycosidases in teir occurrence and properties. PMID:15539

  11. Improved, low-cost selective culture medium for Actinobacillus actinomycetemcomitans.

    Science.gov (United States)

    Alsina, M; Olle, E; Frias, J

    2001-02-01

    Actinobacillus actinomycetemcomitans is considered to be one of the major oral putative pathogens, especially in cases of juvenile periodontitis. This microorganism requires nutritionally complex media for growth, and therefore the media for its primary isolation usually include blood agar or serum in their base. In this study we present a new medium, Dentaid-1, which improves the detection of A. actinomycetemcomitans in periodontal samples. In its composition, blood and serum have been omitted, hence reducing its cost and making it a more restrictive medium against the growth of other microorganisms with high nutritional requirements. The growth yields of pure cultures of the bacteria on Dentaid-1 were comparable to those on nonselective blood agar. Moreover, clinical efficacy was evaluated in subgingival samples from 77 subjects with adult periodontitis. Dentaid-1 detected A. actinomycetemcomitans in 24 subjects, while a previously described tryptic soy-serum-bacitracin-vancomycin agar detected the microorganism in only 19 subjects (79.1%). Dentaid-1 is a low-cost, noninhibitory formula for the improved diagnosis and monitoring of patients subgingivally infected by this important oral putative pathogen.

  12. Inulinase from Kluyveromyces marxianus: culture medium composition and enzyme extraction

    Directory of Open Access Journals (Sweden)

    A. PESSOA JR

    1999-09-01

    Full Text Available K. marxianus DSM 70106 was cultivated for inulinase production in a medium containing 2.0 g/L of yeast extract, 5.0 g/L of peptone and salts. The addition of corn steep liquor did not increase enzyme production. Inulin, as the main carbon source, afforded higher inulinase production than glucose, fructose, sucrose, lactose, maltose and starch. Glucose, fructose and sucrose reduced enzyme production by 46, 58 and 71%, respectively. By using the best culture medium enzyme activity remained stable for 22 months at 4oC; while at -18oC it decreased by 10%. Maximal activity was found in the pH range of 3.5 to 5.0 and at temperatures from 50 to 60oC. Flocculation was used for cell separation. Shifting the pH was more efficient than using polyelectrolytes, CaCl2, bentonite and Fe2O3. Recovery of inulinase by AOT(sodium di-2-ethylhexyl sulfosuccinate-reversed micelles yielded up to ~20%.

  13. M8-An effective medium for anther culture of indica rice

    Institute of Scientific and Technical Information of China (English)

    MEIChuansheng; ZHANGJinyu; WuGuangnan

    1992-01-01

    A new dedifferentiation medium (MS) was developed, which greatly improved the efficiency of anther culture of indica rive. The percentage of green plantlets for anthers inoculated on M8 medium was 40% higher than that on N6 medium in 6 cultivars and it was 2.6%, on average, on M8 medium in more than 20 cultivars and lines,

  14. Mesenchymal stem cell-conditioned medium triggers neuroinflammation and reactive species generation in organotypic cultures of rat hippocampus.

    Science.gov (United States)

    Horn, Ana Paula; Bernardi, Andressa; Luiz Frozza, Rudimar; Grudzinski, Patrícia Bencke; Hoppe, Juliana Bender; de Souza, Luiz Fernando; Chagastelles, Pedro; de Souza Wyse, Angela Terezinha; Bernard, Elena Aida; Battastini, Ana Maria Oliveira; Campos, Maria Martha; Lenz, Guido; Nardi, Nance Beyer; Salbego, Christianne

    2011-07-01

    Cell therapy using bone marrow-derived mesenchymal stem cells (MSCs) seems to be a new alternative for the treatment of neurodegenerative diseases. Despite several promising results with their use, possible side effects are still unknown. In a previous work, we have shown that MSC-conditioned medium is toxic to hippocampal slice cultures and aggravates cell death induced by oxygen and glucose deprivation. In this work, we investigated whether the inflammatory response and/or reactive species formation could be involved in that toxicity. Rat organotypic hippocampal cultures were exposed for 24 h to conditioned medium from MSCs isolated from rat bone marrow. A marked glial activation was observed after exposure of cultures to MSC-conditioned medium, as evidenced by glial fibrillary acid protein (GFAP) and isolectin B(4) increase. Tumor necrosis factor-α and interleukin-6 levels were increased in the culture medium, and 2,7-dihydrodichlorofluorescein diacetate oxidation (indicating reactive species generation) and inducible nitric oxide synthase (iNOS) immunocontent were also higher after exposure of cultures to MSC-conditioned medium. Antioxidants (ascorbic acid and TROLOX(®)), N(ω)-nitro-l-arginine methyl ester hydrochloride, and anti-inflammatory drugs (indomethacin and dexamethasone) reduced cell death in hippocampal organotypic cultures after their exposure to MSC-conditioned medium. The results obtained here suggest that MSC-secreted factors trigger reactive species generation and neuroinflammation in organotypic cultures of hippocampus, introducing a note of caution in the use of these cells for neurological application.

  15. Splitting culture medium by air-jet and rewetting for the assessment of the wettability of cultured epithelial cell surfaces.

    Science.gov (United States)

    Tanaka, Nobuyuki; Kondo, Makoto; Uchida, Ryohei; Kaneko, Makoto; Sugiyama, Hiroaki; Yamato, Masayuki; Okano, Teruo

    2013-12-01

    This study found that the phenomenon of rewetting after squeezing culture medium varied in different culture conditions for rat oral mucosal epithelial cells. When culture medium covering over cultured cells was squeezed by an air-jet application, the motion of squeezed culture medium was able to be observed by using a commercially available movie camera. Squeezed width on cells cultured in keratinocyte culture medium (KCM), which contained with fetal bovine serum, was one-sixth of that in FBS-free KCM. This result corresponded to the mucous layer staining statuses of cultured cells in both cases; positive in KCM and negative in FBS-free medium. Furthermore, the gene expression of mucous glycoprotein MUC4 in KCM was 100 times higher than that in FBS-free medium, and the expression of MUC4 protein only showed on the apical surface of cells cultured in KCM. The relative gene expression levels of MUC1, 13, 15, and 16 in both the normal and FBS-free medium were found to be no more than one-thirtieth of that of MUC4 in KCM. The main factor of the wettability difference between KCM and FBS-free medium was speculated to be the difference of MUC4 expression between both media. This method can be a simple technique for testing not only the surface wettability but also the mucous formation of cultured cells. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. An alternative to evaluate the efficiency of in vitro culture medium using a logistic regression model

    Directory of Open Access Journals (Sweden)

    Daniel Furtado Ferreira

    2003-01-01

    Full Text Available The evaluation of a culture medium for the in vitro culture of a species is performed using its physical and/or chemical properties. However, the analysis of the experimental results makes it possible to evaluate its quality. In this sense, this work presents an alternative using a logistic model to evaluate the culture medium to be used in vitro. The probabilities provided by this model will be used as a medium evaluator index. The importance of this index is based on the formalization of a statistical criterion for the selection of the adequate culture medium to be used on in vitro culture without excluding its physical and/or chemical properties. To demonstrate this procedure, an experiment determining the ideal medium for the in vitro culture of primary explants of Ipeca [Psychotria ipecacuanha (Brot. Stokes] was evaluated. The differentiation of the culture medium was based on the presence and absence of the growth regulator BAP (6-benzilaminopurine. A logistic model was adjusted as a function of the weight of fresh and dry matter. Minimum, medium and maximum probabilities obtained with this model showed that the culture medium containing BAP was the most adequate for the explant growth. Due to the high discriminative power of these mediums, detected by the model, their use is recommended as an alternative to select culture medium for similar experiments.

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

    Directory of Open Access Journals (Sweden)

    S. Sokolov Rossen

    2014-09-01

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

  18. [Identification and antimicrobial susceptibility testing of positive blood culture isolates from briefly incubated solid medium cultures].

    Science.gov (United States)

    Ballestero-Téllez, Mónica; Recacha, Esther; de Cueto, Marina; Pascual, Álvaro

    2016-02-16

    Mass spectrometry Matrix-Assisted Laser Desorption-Ionization Time-of-Flight (MALDI-TOF) helps in the rapid identification of microorganisms causing blood stream infection. Rapid and reliable methods are required to decrease the turnaround time for reporting antimicrobial susceptibility results from blood culture isolates. An evaluation was performed on the reliability of a method for antimicrobial susceptibility testing of positive blood culture isolates from briefly incubated solid medium cultures. The agreement between the evaluated and standard methods was 99.3%. The major and minor error rates were 0.4% and 0.3%, respectively, and no very major errors were observed. The inoculation of briefly incubated solid medium cultures into antimicrobial susceptibility testing panels is an easy and reliable technique, and helps to decrease the turnaround time for reporting antimicrobial susceptibility results of positive blood cultures. Copyright © 2016 Elsevier España, S.L.U. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  19. Generation of large pig and bovine blastocysts by culturing in human induced pluripotent stem cell medium.

    Science.gov (United States)

    Gao, Qing-Shan; Jin, Long; Li, Suo; Zhu, Hai-Ying; Guo, Qing; Li, Xiao-Chen; Jin, Qing-Guo; Kang, Jin-Dan; Yan, Chang-Guo; Yin, Xi-Jun

    2016-04-01

    We investigated the effect of human induced pluripotent stem cell (hiPS) medium on porcine somatic cell nuclear transfer and bovine in vitro fertilized early blastocysts, in comparison with North Carolina State University (NCSU)-37 medium and in vitro culture (IVC)-II medium. After 2 days of culture, the diameter of the portion of the blastocyst that was extruded from the zona pellucid dramatically differed between porcine blastocysts cultured in hiPS medium and those cultured in NCSU-37 medium (221.47 ± 38.94 μm versus 481.87 ± 40.61 μm, P medium and those cultured in IVC-II medium (150.30 ± 29.49 μm versus 195.58 ± 41.59 μm, P cells per porcine and bovine blastocyst was more than two-fold higher in blastocysts cultured in hiPS medium than in those cultured in NCSU-37 medium (44.33 ± 5.28 and 143.33 ± 16.05, P medium (172.12 ± 45.08 and 604.83 ± 242.64, P medium markedly improves the quality of porcine and bovine blastocysts.

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

    Science.gov (United States)

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

    2016-03-01

    Validate single versus sequential culture media for murine embryo development. Prospective laboratory experiment. Assisted Reproduction Laboratory. Murine embryos. Thawed murine zygotes cultured for 3 or 5 days (d3 or d5) in single or sequential embryo culture media developed for human in vitro fertilization. On d3, zygotes developing to the 8 cell (8C) stage or greater were quantified using 4',6-diamidino-2-phenylindole (DAPI), and quality was assessed by morphological analysis. On d5, the number of embryos reaching the blastocyst stage was counted. DAPI was used to quantify total nuclei and inner cell mass nuclei. Localization of ubiquitin C-terminal hydrolase L1 (UCHL1) and ubiquitin C-terminal hydrolase L3 (UCHL3) was reference points for evaluating cell quality. Comparing outcomes in single versus to sequential media, the odds of embryos developing to the 8C stage on d3 were 2.34 time greater (P = .06). On d5, more embryos reached the blastocyst stage (P = quality hatching blastocysts. Localization of UCHL1 and UCHL3 was diffuse and inappropriately dispersed throughout the cytoplasm in low-quality nonhatching blastocysts. Single medium yields greater cell numbers, an increased growth rate, and more hatching of murine embryos. Cytoplasmic UCHL1 and nuclear UHCL3 localization patterns were indicative of embryo quality. Our conclusions are limited to murine embryos but one might speculate that single medium may also be more beneficial for human embryo culture. Human embryo studies are needed. © The Author(s) 2015.

  1. Cultural Studies and Media Ecology: Meyrowitz's Medium Theory and Carey's Cultural Studies.

    Science.gov (United States)

    Flayhan, Donna P.

    2001-01-01

    Examines work of two communication and media studies scholars, Joshua Meyrowitz and James Carey. Suggests their studies represent media ecology with analyses of the dynamic interaction between communication, consciousness, and culture. Highlights how their work embodies a North American cultural studies approach to media studies (moving away from…

  2. Cultural Studies and Media Ecology: Meyrowitz's Medium Theory and Carey's Cultural Studies.

    Science.gov (United States)

    Flayhan, Donna P.

    2001-01-01

    Examines work of two communication and media studies scholars, Joshua Meyrowitz and James Carey. Suggests their studies represent media ecology with analyses of the dynamic interaction between communication, consciousness, and culture. Highlights how their work embodies a North American cultural studies approach to media studies (moving away from…

  3. Long-term in vitro culture of bovine preantral follicles: Effect of base medium and medium replacement methods.

    Science.gov (United States)

    Araújo, V R; Gastal, M O; Wischral, A; Figueiredo, J R; Gastal, E L

    2015-10-01

    Two culture media and replacement methods were compared during long-term in vitro culture of secondary follicles of cattle using α-MEM(+) or TCM-199(+) as base media. The medium replacement methods were: Conventional - removal and subsequent addition of the same amount (60μl) in a 100μl aliquot (MEM-C and TCM-C), and Small Supplementation - addition of 5μl of fresh medium to an initial small aliquot (50μl), resulting in a final volume of 125μl on the last day of culture (MEM-S and TCM-S). A total of 207 secondary follicles were cultured individually for 32 days at 38.5°C in 5% CO2 and medium replacement was performed every other day. The MEM-S treatment resulted in a larger (P0.05) the follicular and estradiol end points for TCM-199(+). The expression of the FSHR gene was greater (Pculture of preantral follicles of cattle if progressive addition of medium is used for medium change.

  4. Characterization of Residual Medium Peptides from Yersinia pestis Cultures

    Energy Technology Data Exchange (ETDEWEB)

    Clowers, Brian H.; Wunschel, David S.; Kreuzer, Helen W.; Engelmann, Heather E.; Valentine, Nancy B.; Wahl, Karen L.

    2013-04-03

    Using a range of common microbial medium formulations (TSB, BHI, LB, and G-media), two attenuated strains of Y. pestis (KIM D27 (pgm-) and KIMD1 lcr-) were cultivated in triplicate. These cellular suspensions were used to develop a method of extracting residual medium peptides from the final microbial preparation to assess their relative abundance and identity. Across the conditions examined, which included additional cellular washing and different forms of microbial inactivation, residual medium peptides were detected. Despite the range of growth medium sources used and the associated manufacturing processes used in their production, a high degree of peptide similarity was observed for a given medium recipe. These results demonstrate that residual medium peptides are retained using traditional microbial cultivation techniques and may be used to inform forensic investigations with respect to production deduction.

  5. Selection of medium for serum-free primary culture of adult rat hepatocytes.

    Directory of Open Access Journals (Sweden)

    Miyazaki,Masahiro

    1990-02-01

    Full Text Available To select a suitable medium for serum-free primary culture of adult rat hepatocytes, ten commercially-available synthetic media were compared for their ability to maintain the cells under serum-free and serum-supplemented conditions with special reference to attachment, survival and albumin secretion. It was found that Williams' medium E and DM-160 medium were the best among the ten media for maintaining hepatocytes under serum-free conditions in primary culture.

  6. Culture medium refinement by dialysis for the expansion of human induced pluripotent stem cells in suspension culture.

    Science.gov (United States)

    Nath, Suman Chandra; Nagamori, Eiji; Horie, Masanobu; Kino-Oka, Masahiro

    2017-01-01

    Human induced pluripotent stem cells (hiPSCs) secrete essential autocrine factors that are removed along with toxic metabolites when the growth medium is exchanged daily. In this study, after determining the minimum inhibitory level of lactic acid for hiPSCs, a medium refining system was constructed by which toxic metabolites were removed from used culture medium and autocrine factors as well as other growth factors were recycled. Specifically, about 87 % of the basic fibroblast growth factor and 80 % of transforming growth factor beta 1 were retained in the refined medium after dialysis. The refined medium efficiently potentiated the proliferation of hiPS cells in adherent culture. When the refining system was used to refresh medium in suspension culture, a final cell density of (1.1 ± 0.1) × 10(6) cells mL(-1) was obtained, with 99.5 ± 0.2 % OCT 3/4 and 78.3 ± 1.1 % TRA-1-60 expression, on day 4 of culture. These levels of expression were similar to those observed in the conventional suspension culture. With this method, culture medium refinement by dialysis was established to remove toxic metabolites, recycle autocrine factors as well as other growth factors, and reduce the use of macromolecules for the expansion of hiPSCs in suspension culture.

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

    Directory of Open Access Journals (Sweden)

    Mazur Jolanta

    2016-09-01

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

  8. The effect of medium composition on ovary-slice culture and ovule culture in intraspecific Tulipa gesneriana crosses

    NARCIS (Netherlands)

    Creij, van M.G.M.; Kerckhoffs, D.M.F.J.; Bruijn, de S.M.; Tuyl, van J.M.; Vreugdenhil, D.

    2000-01-01

    The effect of several media components on the germination percentage of ovules in intraspecific T. gesneriana L. crosses was studied by using two embryo rescue techniques, viz. ovary-slice culture followed by ovule culture and direct ovule culture. The addition of 9% sucrose to medium for

  9. Glucose Levels in Culture Medium Determine Cell Death Mode in MPP(+)-treated Dopaminergic Neuronal Cells.

    Science.gov (United States)

    Yoon, So-Young; Oh, Young J

    2015-09-01

    We previously demonstrated that 1-methyl-4-phenylpyridinium (MPP(+)) causes caspase-independent, non-apoptotic death of dopaminergic (DA) neuronal cells. Here, we specifically examined whether change of glucose concentration in culture medium may play a role for determining cell death modes of DA neurons following MPP(+) treatment. By incubating MN9D cells in medium containing varying concentrations of glucose (5~35 mM), we found that cells underwent a distinct cell death as determined by morphological and biochemical criteria. At 5~10 mM glucose concentration (low glucose levels), MPP(+) induced typical of the apoptotic dell death accompanied with caspase activation and DNA fragmentation as well as cell shrinkage. In contrast, MN9D cells cultivated in medium containing more than 17.5 mM (high glucose levels) did not demonstrate any of these changes. Subsequently, we observed that MPP(+) at low glucose levels but not high glucose levels led to ROS generation and subsequent JNK activation. Therefore, MPP(+)-induced cell death only at low glucose levels was significantly ameliorated following co-treatment with ROS scavenger, caspase inhibitor or JNK inhibitor. We basically confirmed the quite similar pattern of cell death in primary cultures of DA neurons. Taken together, our results suggest that a biochemically distinct cell death mode is recruited by MPP(+) depending on extracellular glucose levels.

  10. Culture medium optimization for pigment production with RSM method

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    Response surface methodology was used to optimize a medium for a red-pigmented marine bacterium S-9801 strain (Flavobacterium sp.). In the first optimization step the influence of yeast extract, peptone, glucose and sodium chloride on pigment production was evaluated using a fractional factorial design. Pigment production was positively influenced by glucose and sodium chloride while other components had no significant effect. In the second step the path of steepest ascent was used to approach the optimal region of the medium composition. In the third step the optimal concentration of glucose and sodium chloride was determined by a central composite design and response analysis. The optimized medium allowed pigment production (A 535~650) to be increased from 0.137 to0.559, being 320% higher than the original medium.

  11. Culturing in serum-free culture medium on collagen type-I-coated plate increases expression of CD133 and retains original phenotype of HT-29 cancer stem cell.

    Science.gov (United States)

    Arab-Bafrani, Zahra; Shahbazi-Gahrouei, Daryoush; Abbasian, Mehdi; Saberi, Alihossein; Fesharaki, Mehrafarin; Hejazi, Seyed Hossein; Manshaee, Samira

    2016-01-01

    A sub-population of tumor cells termed cancer stem cells (CSCs) has an important role in tumor initiation, progression, and recurrence. Selecting a suitable procedure for isolation and enrichment of CSCs is the biggest challenge in the study of CSCs. In the present study, the role of the combination of stem cell culture medium and collagen type-I was evaluated for successful isolation and enrichment of HT-29 CSCs. HT-29 cells were cultured in serum-containing medium (parental culture medium: Medium + 10% fetal bovine serum) and serum-free medium (stem cell culture medium); both on collagen-coated plates. Spheres forming ability and CD133 expression, as a potential marker of colorectal CSCs, were evaluated in two culture mediums. The results show spheroids usually give rise completely within 15 days in the stem cell culture medium on the collagen-coated plate. CD133 expression in spheroid cells (84%) is extensively higher than in parental cells (25%). Moreover, relative to parental cells, spheroid cells were more radioresistance. Finding of this study suggested that CSCs derived from colon cancer cell line (HT-29) can be propagated and form colonospheres in serum-free culture medium on collagen type-I. According to maintenance of their original phenotype in these conditions, it seems serum-free culture medium on collagen type-I is a suitable way to drug screening of HT-29 CSCs.

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

    KAUST Repository

    Hadj-Romdhane, F.

    2013-03-01

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

  13. [Method of producing fusarin C in perlite-liquid culture medium].

    Science.gov (United States)

    Li, G; Li, M; Ma, J

    1992-02-01

    For researching the biosynthesis labelled Fusarin C(Fc) by Fuscarium moniliforme, a more quick and convenient method of Fusarin C production and purification were established, and a good liquid culture medium consisted of different kinds organic matters (hydroxy proline, sucrose and glycerin), inorganic salts and perlite replaced corn grit medium. The perlite-liquid culture medium inoculated with the strain of F. moniliforme yields 936mg Fc/kg organic matter with in 14 days of incubation at 28 degrees C. As compared with the corn grit medium, the amount of Fc from perlite-liquid medium was more than that from corn grit medium (831mg Fc/kg corn grit). In all experiments both thin-layer chromatography and high-pressure liquid chromatography were used to confirm the presence of Fc. parameters which were important for the optimal biosynthesis of Fc included hydroxy proline and sucrose concentrations, incubated time/temperature and amount of perlite. The 40g of sucrose/L liquid culture was optimal concentration for Fusarin C production. Of three contained N-matter tested, hydroxy proline was the best sources of N-atom for Fusarin C. Under the absence of hydroxy proline, the Fc wasn't synthesized in perlite-liquid culture medium by F. moniliforme. A culture time/temperature study of Fc production was done, and the optimal Fc amounts was synthesized after incubation for 14 days at 28 degrees C on perlite-liquid culture medium.

  14. Design of serum-free medium for suspension culture of CHO cells on the basis of general commercial media.

    Science.gov (United States)

    Miki, Hideo; Takagi, Mutsumi

    2015-08-01

    The design of serum-free media for suspension culture of genetically engineered Chinese hamster ovary (CHO) cells using general commercial media as a basis was investigated. Subcultivation using a commercial serum-free medium containing insulin-like growth factor (IGF)-1 with or without FCS necessitated additives other than IGF-1 to compensate for the lack of FCS and improve cell growth. Suspension culture with media containing several combinations of growth factors suggested the effectiveness of addition of both IGF-1 and the lipid signaling molecule lysophosphatidic acid (LPA) for promoting cell growth. Subcultivation of CHO cells in suspension culture using the commercial serum-free medium EX-CELL™302, which contained an IGF-1 analog, supplemented with LPA resulted in gradually increasing specific growth rate comparable to the serum-containing medium and in almost the same high antibody production regardless of the number of generations. The culture with EX-CELL™302 supplemented with LPA in a jar fermentor with pH control at 6.9 showed an apparently higher cell growth rate than the cultures without pH control and with pH control at 6.8. The cell growth in the medium supplemented with aurintricarboxylic acid (ATA), which was much cheaper than IGF-1, in combination with LPA was synergistically promoted similarly to that in the medium supplemented with IGF-1 and LPA. In conclusion, the serum-free medium designed on the basis of general commercial media could support the growth of CHO cells and antibody production comparable to serum-containing medium in suspension culture. Moreover, the possibility of cost reduction by the substitution of IGF-1 with ATA was also shown.

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

    Science.gov (United States)

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

    2013-09-01

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

  16. Culture Teaching in China’s Current College English Class and Some Suggestions

    Institute of Scientific and Technical Information of China (English)

    李思远

    2014-01-01

    Culture is an important part in foreign language learning because no language can leave its culture contexts. Therefore, if one wants to learn a foreign language well, the acculturation is very important. But in the Chinese EFL classroom, for kinds of reasons, teachers often pay more attention to the explanation of grammatical rules and ignored the culture input which makes many Chinese students lack of pragmatic competence and can not use English in a proper way. This article will analyze the cur-rent culture teaching in our college English class and propose some suggestions.

  17. [Culture medium optimization and primary kinetics analysis for nisin production].

    Science.gov (United States)

    Li, C; Bai, J H; Cai, Z L; Ouyang, F

    2001-03-01

    Response surface methodology was used to optimize a medium for nisin production of Lactococcus lactis. In the first optimization step the influence of sucrose, soybean peptone, yeast extract, potassium dihydrogen phosphate, sodium chloride, and magnesium sulfur on nisin production was evaluated using a fractional factorial design. Potassium dihydrogen phosphate influenced nisin production positively while soybean peptone affected nisin production negatively. The other components had no significant effect on nisin production. The path of steepest ascent was used to approach the optimal region of the medium composition. In the third step the optimal concentrations of KH2PO4 and soybean peptone were determined by a central composite design and response surface analysis. The optimized medium allowed nisin production to be increased from 1074 IU/mL to 2150 IU/mL. The kinetic analysis showed that nisin production fashion at optimized and non-optimized media was not changed and maintained partially growth-associated. But the specific growth rates and the specific nisin production rates for the strain at the optimized medium were bigger than the ones at the non-optimized medium after the cells entered the middle of exponential phase.

  18. Culture of proliferating and differentiating fat-storing cells in 3T3-conditioned medium.

    Science.gov (United States)

    Mendoza-Figueroa, T; Argüello, C; Kuri-Harcuch, W

    1988-01-01

    There is growing evidence suggesting that hepatic fat-storing cells (FSC) or Ito cells have an important function in vitamin A storage and metabolism and in the synthesis of connective tissue components in normal liver and during fibrogenesis. The purified FSC acquire a fibroblastic morphology and their vitamin A content decreases in culture. We cultivated cells under in vitro conditions that allowed the expression of FSC morphological and functional characteristics for 3-4 weeks of primary culture. Cells were isolated from rat liver by the collagenase-perfusion method without further purification and cultured with 3T3-conditioned medium, which seemed to stimulate the selective proliferation of the FSC. After 8-10 days, round and stellate cells grew actively from a few precursor cells in the primary culture and were not subcultivated; the stellate cells had the ability to become round and vice versa and were highly motile. The cells had intracytoplasmic lipid droplets, a well developed rough endoplasmic reticulum, Golgi complex, numerous vesicles filled with electron-dense material, and extracellular matrix (ECM) components on their surface. Both stellate and round cells showed the presence of desmin by immunofluorescence and vitamin A autofluorescence, but lacked peroxidase activity. The culture conditions we describe allowed the selective proliferation of cells with morphological and functional characteristics of the FSC in the normal liver, raising the possibility of studying FSC proliferation and differentiation.

  19. A Study on the Relationship between Organizational Culture and Organizational Performance and a Model Suggestion

    Directory of Open Access Journals (Sweden)

    Ebru Yildiz

    2016-01-01

    Full Text Available Organizational culture is defined as set of goals and values shared by employees in the organization. Recently it has also come to be perceived as a resource of knowledge in the organization. In the literature, along with the studies aiming at identifying the elements of organizational culture, there are some research that study the relationship between organizational culture and performance. This article is a theoretical study of the two concepts and suggests a research model for further research. Recently, knowledge management and innovation strategy have started to be regarded as features of organizational culture. Research shows that these two variables also have significant impacts on organizational performance. Our model suggests measuring the effect of organizational culture on performance along with the supportive impacts of knowledge management and innovation strategy.

  20. IN-VITRO CULTURE FOR REGENERATION OF Melia azedarach L. USING AGITATED LIQUID MEDIUM

    Directory of Open Access Journals (Sweden)

    Arif Nirsatmanto

    2006-07-01

    Full Text Available This study investigated the applicability of liquid medium in direct organogenesis of in-vitro culture of Melia azedarach L. Explants were collected from in-vitro multiplication originating from aseptically germinated seedling and from a 48 - years old mature tree. For adventitious shoot differentiation, 2 mm length of excised explants were cultured on liquid medium of MS (Murashige and Skoog's basal medium supplemented with 28 combinations of hormone BAP (6-benzylaminopurine and NAA (á- naphthaleneacetic acid. Differentiated shoots were subsequently cultured for shoot elongation in solid medium using MS basal medium supplemented with hormone BAP individually as well as in combinations with NAA. Finally, rooting culture were done using MS medium supplemented with IBA (3  indolebutyric acid. The results showed that the rate of shoot organogenesis of M. azedarach could be obtained through agitated liquid medium culture technique. The combination of hormone BAP 0.1-1 µM and NAA 0.01-0.1 µM are induced more adventitious shoot at a rate of 5 shoots / 2 mm length size of explants are transferring into rooting medium containing IBA 4.92 µM.

  1. [Effect of calcium on medium alkalinization induced by salicylic acid in Salvia miltiorrhiza suspension cultures].

    Science.gov (United States)

    Liu, Liancheng; Wang, Cong; Dong, Juan'e; Su, Hui; Zhuo, Zequn; Xue, Yaxin

    2013-07-01

    We studied medium alkalinization in Salvia miltiorrhiza suspension cultures treated with salicylic acid and the effect of Ca2+ in this process through application of calcium channel antagonists (Verapamil, LaCl3, LiCl, 2-APB) and ionophore A23187. The results show that salicylic acid could induce significant medium alkalinization in S. miltiorrhiza culture. Verapamil and LaCl3 or LiCl and 2-APB, two different groups of calcium channel antagonist, significantly inhibited the medium alkalinization induced by salicylic acid. However, the suppression effect of verapamil or LaCl3 on medium alkalinization induced by salicylic acid was higher than that of LiCl or 2-APB. When two types of calcium channel inhibitor (LaCl3 and 2-APB) were used together, the medium alkalinization induced by salicylic acid was completely suppressed and even reduced the pH in medium. On the other hand, A23187 could promote the medium alkalinization. Based on the results above, we speculated that salicylic acid could induce significant medium alkalinization in S. miltiorrhiza culture, depending on the calcium from both extracell and intracell. Moreover, calcium from extracell plays a more dominant role in this process. Reveal of relationship in this research between Ca2+ and medium alkalinization can provide theory evidence for mechanism of the plant secondary metabolism.

  2. Growth and maintenance of an embryogenic cell culture of daylily (Hemerocallis) on hormone-free medium

    Science.gov (United States)

    Smith, D. L.; Krikorian, A. D.

    1991-01-01

    Callus cultures of the diploid daylily (Hemerocallis) clone Autumn Blaze' were initiated and maintained in hormone-containing nutrient medium. At various times (from 6 weeks to 1 year) after being initiated, hormone-derived cultures were evaluated for their ability to be maintained and to multiply on hormone-free medium at low pH (between pH 4 and 4.5). Cultures had to be exposed to hormone-containing medium for at least 12 weeks before they could be maintained on hormone-free medium at low pH. The transition to maintainability on low pH hormone-free medium included the production of many aberrant embryonal forms ( neomorphs'). However, all hormone-derived cultures tested consisted entirely of preglobular stage proembryos (PGSPs) after 12-24 weeks on low pH hormone-free medium. PGSP cultures have been maintained and multiplied as such for over 1 year on low pH hormone-free medium. PGSPs continue their development into various somatic embryo stages when cultured on hormone-free medium buffered at pH 5.8. The production of well-formed somatic embryos was greatly enhanced when PGSPs were plated on activated charcoal impregnated filter papers that were placed on top of the agar surface. The gross morphology and histology of the PGSPs and stages of somatic embryo development are presented. The work shows that the ability of hormone-free medium at low pH to permit PGSP multiplication without development into later stages of embryo development is not restricted to carrot.

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

    Directory of Open Access Journals (Sweden)

    Silvio Lopes Teixeira

    2009-10-01

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

  4. Viability and growth of feline preantral follicles in vitro cultured with insulin growth factor and epidermal growth factor supplemented medium.

    Science.gov (United States)

    Alves, A E; Padilha-Nakaghi, L C; Pires-Butler, E A; Apparicio, M; Silva, Nam; Motheo, T F; Vicente, Wrr; Luvoni, G C

    2017-04-01

    In vitro culture of ovarian preantral follicles has emerged as a reproductive technology aimed at obtaining large amount of oocytes for in vitro embryo production. The addition of growth factors (GF) in the in vitro culture of preantral follicles of different species has provided superior results of follicular development, antrum formation and proliferation of granulosa cells. However, there are only few reports regarding the use of these factors on feline preantral follicle in vitro culture. Thus, the aim of this study was to investigate the effect of a combination of IGF-1 and EGF on in vitro viability and growth of preantral follicles and enclosed oocytes collected from domestic cats. A total of 64 follicles characterized by multilayer granulosa cells were isolated and individually cultured for 6 days (T6) in minimum essential medium supplemented with IGF-1+ EGF (100 ng/ml each) or without (control). A higher percentage of follicles were viable after culture with GF than without, and an increase in size when IGF-1+ EGF were added to the medium (170 ± 32.4 μm (T0) vs. 201 ± 22.3 μm (T6); p  .05). These data suggest that the addition of IGF-1 and EGF to the culture medium promotes the in vitro development of preantral follicles of cats. © 2016 Blackwell Verlag GmbH.

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

    Directory of Open Access Journals (Sweden)

    Villegas E Valeska

    2007-12-01

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

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

    NARCIS (Netherlands)

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

    2016-01-01

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

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

    NARCIS (Netherlands)

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

    2015-01-01

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

  8. Coculture of osteoblasts and endothelial cells: optimization of culture medium and cell ratio

    NARCIS (Netherlands)

    Ma, J.; Beucken, J.J. van den; Yang, F.; Both, S.K.; Cui, F.Z.; Pan, J.; Jansen, J.A.

    2011-01-01

    Vascularization strategies in cell-based bone tissue engineering depend on optimal culture conditions. The present study aimed to determine optimal cell culture medium and cell ratio for cocultures of human marrow stromal cells (HMSCs) and human umbilical vein endothelial cells (HUVECs) in view of

  9. Modified PEHPS Medium as an Alternative for the In Vitro Culture of Giardia lamblia

    Science.gov (United States)

    Vargas-Villarreal, Javier; Mata-Cárdenas, Benito D.; Hernández-García, Magda E.; Garza-González, Jesús N.; De La Garza-Salinas, Laura H.; González-Salazar, Francisco

    2014-01-01

    Commercial culture media present interlot variations in biological activity. We have previously designed a homemade and economic culture medium, PEHPS medium, for the axenic cultivation of Entamoeba histolytica and Trichomonas vaginalis. Trophozoites of amoebae and trichomonads grow well in this medium. Furthermore, the medium is stable for several months when stored frozen or refrigerated. The objective of this work was to modify PEHPS medium to support the in vitro growth of Giardia lamblia. Inocula of 5 × 103 trophozoites/mL of G. lamblia were incubated at 36.5°C in modified PEHPS or TYI-S-33 medium. Then, the growths of the three Giardia strains in both media were compared. The logarithmic growth phase lasted 72 h; the mean yield of the strains ranged from 10.06 to 11.43 × 105Giardia trophozoites/mL, and the range of duplication time in the three strains was from 5.67 to 6.06 in modified PEHPS medium. These growth characteristics were not significantly different from those obtained with TYI-S-33 medium. We conclude that modified PEHPS medium might be used for the axenic cultivation of G. lamblia. PMID:24982905

  10. Modified PEHPS medium as an alternative for the in vitro culture of Giardia lamblia.

    Science.gov (United States)

    Vargas-Villarreal, Javier; Mata-Cárdenas, Benito D; Hernández-García, Magda E; Garza-González, Jesús N; De La Garza-Salinas, Laura H; González-Salazar, Francisco

    2014-01-01

    Commercial culture media present interlot variations in biological activity. We have previously designed a homemade and economic culture medium, PEHPS medium, for the axenic cultivation of Entamoeba histolytica and Trichomonas vaginalis. Trophozoites of amoebae and trichomonads grow well in this medium. Furthermore, the medium is stable for several months when stored frozen or refrigerated. The objective of this work was to modify PEHPS medium to support the in vitro growth of Giardia lamblia. Inocula of 5 × 10(3) trophozoites/mL of G. lamblia were incubated at 36.5°C in modified PEHPS or TYI-S-33 medium. Then, the growths of the three Giardia strains in both media were compared. The logarithmic growth phase lasted 72 h; the mean yield of the strains ranged from 10.06 to 11.43 × 10(5) Giardia trophozoites/mL, and the range of duplication time in the three strains was from 5.67 to 6.06 in modified PEHPS medium. These growth characteristics were not significantly different from those obtained with TYI-S-33 medium. We conclude that modified PEHPS medium might be used for the axenic cultivation of G. lamblia.

  11. Modified PEHPS Medium as an Alternative for the In Vitro Culture of Giardia lamblia

    Directory of Open Access Journals (Sweden)

    Javier Vargas-Villarreal

    2014-01-01

    Full Text Available Commercial culture media present interlot variations in biological activity. We have previously designed a homemade and economic culture medium, PEHPS medium, for the axenic cultivation of Entamoeba histolytica and Trichomonas vaginalis. Trophozoites of amoebae and trichomonads grow well in this medium. Furthermore, the medium is stable for several months when stored frozen or refrigerated. The objective of this work was to modify PEHPS medium to support the in vitro growth of Giardia lamblia. Inocula of 5 × 103 trophozoites/mL of G. lamblia were incubated at 36.5°C in modified PEHPS or TYI-S-33 medium. Then, the growths of the three Giardia strains in both media were compared. The logarithmic growth phase lasted 72 h; the mean yield of the strains ranged from 10.06 to 11.43 × 105Giardia trophozoites/mL, and the range of duplication time in the three strains was from 5.67 to 6.06 in modified PEHPS medium. These growth characteristics were not significantly different from those obtained with TYI-S-33 medium. We conclude that modified PEHPS medium might be used for the axenic cultivation of G. lamblia.

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

    Science.gov (United States)

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

    2012-06-22

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

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

    Science.gov (United States)

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

    2016-03-01

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

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

    Directory of Open Access Journals (Sweden)

    M. DANCI

    2013-12-01

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

  15. Culture medium optimization for osmotolerant yeasts by use of a parallel fermenter system and rapid microbiological testing.

    Science.gov (United States)

    Pfannebecker, Jens; Schiffer-Hetz, Claudia; Fröhlich, Jürgen; Becker, Barbara

    2016-11-01

    using OM in combination with the automated test system even if low initial counts (10(1)CFU/g) are present in the products. In conclusion, the presented data suggest that the OM culture medium is appropriate for the enrichment of osmotolerant yeasts from high-sugar food products. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Solid Medium for Culturing Black Smoker Bacteria at Temperatures to 120°C

    OpenAIRE

    Deming, Jody W.; Baross, John A.

    1986-01-01

    A solid, highly thermostable medium, based on the new gelling agent GELRITE, was devised to facilitate the culturing of extremely thermophilic microorganisms from submarine hydrothermal vents. The medium remained solid at temperatures to 120°C at vapor pressures and hydrostatic pressures to 265 atm. It proved useful to its maximum tested limits in isolating colonies of black smoker bacteria from hydrothermal fluids recently collected at the Juan de Fuca Ridge in the Pacific Ocean.

  17. Cartilage storage at 4 °C with regular culture medium replacement benefits chondrocyte viability of osteochondral grafts in vitro.

    Science.gov (United States)

    Qi, Jianhong; Hu, Zunjie; Song, Hongqiang; Chen, Bin; Xie, Di; Zhou, Lu; Zhang, Yanming

    2016-09-01

    Maintenance of articular cartilage allografts in culture media is a common method of tissue storage; however, the technical parameters of graft storage remain controversial. In this study, we examined the optimal temperature and culture medium exchange rate for the storage of osteochondral allografts in vitro. Cylindrical osteochondral grafts (n = 120), harvested from the talar joint surface of ten Boer goats, were randomly classified into four groups and stored under the following conditions: Group A1 was maintained at 4 °C in culture medium that was refreshed every 2 days; Group A2 was maintained at 4 °C in the same culture medium, without refreshing; Group B1, was maintained at 37 °C in culture medium that was refreshed every 2 days; Group B2, was maintained at 37 °C in the same culture medium, without refreshing. Chondrocyte viability in the grafts was determined by ethidium bromide/fluorescein diacetate staining on days 7, 21, and 35. Proteoglycan content was measured by Safranin-O staining. Group A1 exhibited the highest chondrocyte survival rates of 90.88 %, 88.31 % and 78.69 % on days 7, 21, and 35, respectively. Safranin O staining revealed no significant differences between groups on days 21 and 35. These results suggest that storage of osteochondral grafts at 4 °C with regular culture medium replacement should be highly suitable for clinical application.

  18. Evaluation of Endotoxin in Culture Medium for Human in vitro Fertilization

    Institute of Scientific and Technical Information of China (English)

    Jing LI; Wei-jie ZHU; Wen-hong ZHANG

    2005-01-01

    Objective To determine whether the presence of bacterial endotoxin in the commercial culture media utilized for human in vitro fertilization (IVF), and evaluate the difference in detecting endotoxin in culture medium between the human sperm motility assay and the 2-cell mouse embryo assay.Methods Thirty-six batches of culture media commonly used in IVF laboratories from 3 manufacturers were determined for the presence ofendotoxin before using the medium for the assisted reproductive programs (group A). After being used, 25 specimens among above media were also tested (group B). The chromogenic limulus amoebocyte lysate (LAL) test was used for quantification the content of endotoxin. In addition, the human sperm motility assay was compared with the 2-cell mouse embryo assay to evaluate the difference in detecting endotoxin in culture medium.Results Endotoxin was not detected in group A. However, 2 samples were positive in group B. Sperm did not show significant change in motility in group A during 24 h of incubation when compared with the control (P>0.05). However, in group A the 2-cell embryo development to blastocyst was suppressed in 3 batches of media.Conclusions Regular screening of each batch of culture medium should be performed if possible although there was no evidence of endotoxin contamination in commercially prepared pre-tested media. Culture environment should be stringently controlled in case the medium is polluted. The sensitivity of the sperm motility assay was lower than that of the mouse embryo assay for detecting low levels of endotoxin or toxic compounds in the medium.

  19. Integrating insulin into single-step culture medium regulates human embryo development in vitro.

    Science.gov (United States)

    Fawzy, Mohamed; Sabry, Mohamed; Nour, Mohamed; Abdelrahman, Mohamed Y; Roshdy, Eman; Magdi, Yasmin; Abdelghafar, Hazem

    2017-02-01

    To evaluate the effect of supplementing single-step embryo culture medium with insulin on human embryo development. Comparative study. Two private centers. The study involved a sibling oocyte split of 5,142 retrieved oocytes from 360 patients. Sibling oocytes split after intracytoplasmic sperm injection for culture from day 0 through day 5 or 6 in insulin-supplemented or control medium. Women were split to receive their embryos from insulin-supplemented or control medium. Clinical pregnancy rate. There were significantly higher rates of clinical, ongoing, and twin pregnancies in the insulin-supplemented arm than in the control arm. On day 3, embryo quality and compaction were higher in insulin-supplemented medium. On day 5, insulin supplementation showed higher rates of blastocyst formation, quality, and cryopreservation. Insulin supplementation of single-step embryo culture medium from day 0 through day 5 or 6 improved clinical pregnancy rate and human embryo development. However, these findings need further confirmation through a multicenter randomized controlled trial that may include other patient populations and different culture media. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  20. Influence of medium composition on oxygen transfer rate in animal cell culture

    OpenAIRE

    Toye, Dominique; Galifi, Alain; Salmon, Thierry; Verdin, Emeline; Crine, Michel

    2009-01-01

    Experiments were conducted in a 0.25 m diameter bubble column to investigate the effect of animal cell culture medium composition on oxygen transfer rate. Air is used as the dispersed phase. The gas superficial velocity is varied between 0.4 and 2 cm/s (aeration rate ranging between 0.05 and 0.25 vvm) and the bubble column is thus operated in the homogenous regime. Aqueous solutions the composition (electrolyte, protein concentrations) of which mimics a mammalian cell culture medium) are us...

  1. Culture medium-associated physicochemical insights on the cytotoxicity of carbon nanomaterials.

    Science.gov (United States)

    Kong, Huating; Wang, Lihua; Zhu, Ying; Huang, Qing; Fan, Chunhai

    2015-03-16

    Carbon nanomaterials are the most studied materials in nanotechnology. There have been numerous studies on cytotoxicity assessments of carbon nanomaterials, which, however, often lead to controversy. It is generally considered that chemical and physical properties of carbon nanomaterials should have specific biological outcomes. More recent studies have identified the significance of environmental factors surrounding nanomaterial-treated cells. In this perspective, we mainly review culture medium-associated physicochemical insights on the cytotoxicity of carbon nanomaterials, which are largely based on studies in our laboratory. These studies established the close relationship and interplay among the physicochemical properties of the nanomaterials, culture medium, and their toxicological responses.

  2. Culturing in serum-free culture medium on collagen type-I-coated plate increases expression of CD133 and retains original phenotype of HT-29 cancer stem cell

    Directory of Open Access Journals (Sweden)

    Zahra Arab-Bafrani

    2016-01-01

    Conclusion: Finding of this study suggested that CSCs derived from colon cancer cell line (HT-29 can be propagated and form colonospheres in serum-free culture medium on collagen type-I. According to maintenance of their original phenotype in these conditions, it seems serum-free culture medium on collagen type-I is a suitable way to drug screening of HT-29 CSCs.

  3. [Culture medium based on biogas slurry and breeding of oil Chlorella].

    Science.gov (United States)

    Zhao, Feng-Min; Mei, Shuai; Cao, You-Fu; Ding, Jin-Feng; Xu, Jia-Jie; Li, Shu-Jun

    2014-06-01

    The oil chlorella cultivation and biogas slurry treatment were combined. The biogas slurry provided water and nutrient for growing chlorella, at the same time, harmless treatment of biogas slurry was realized. This paper cultivated 4 species of oil chlorella in the mixed medium of biogas slurry and green algae medium (the volume ratios were 1 : 9, 1 : 3, 1 : 1 and 3 : 1, respectively), and compared their oil productivity to select the best oil chlorella species and the optimal culture medium. The results showed that, the combination of medium and chlorella species to reach the highest oil productivity was a volume ratio of 1 : 3 and the chlorella species BJ05, and the oil productivity of chlorella BJ05 was 9.20 mg x (L x d)(-1), higher than that in green algae medium [8.66 mg x (L x d)(-1)]. In mixed medium with a volume ratio of 1:3, the effect of adding different nutrients into the green algae medium on the oil productivity was examined, and the results showed that, sodium carbonate and citric acid had no negative effect on the oil productivity of chlorella BJ05. in the absence of sodium carbonate and citric acid, the oil productivity of chlorella BJ05 was 9.36 mg x (L x d)(-1), and the removal of COD (chemical oxygen demand), total nitrogen, total phosphorus and ammonia nitrogen rates were 59%, 75%, 61% and 100%, respectively. Deficiency in other nutrients had negative effect on the oil productivity. Therefore, the culture medium was further optimized to the mixed medium of biogas slurry and green algae medium with a volume ratio of 1 : 3 and without addition of sodium carbonate and citric acid.

  4. Cd-induced phytochelatin synthesis in Dittrichia viscosa (L.) Greuter is determined by the dilution of the culture medium.

    Science.gov (United States)

    Fernández, R; Fernández-Fuego, D; Rodríguez-González, P; Alonso, J I García; Bertrand, A; González, A

    2014-01-01

    In this paper, we examined Cd accumulation and PC synthesis in two clones of Dittrichia viscosa, one with a metallicolous (DV-A) and the other with a non-metallicolous origin (DV-W). The clones were cultured in vitro with 0 and 10 mg Cd L(-1) in both short-term treatments (up to 72 h) and over 10 days. We also examined the influence of the culture medium dilution and the PC-synthesis inhibitor, L-buthionine-sulfoximine (BSO), on these parameters. Similar Cd accumulation values were found in the two clones. No synthesis of new thiolic compounds was observed in Cd-treated plants cultured in vitro in Murashige and Skoog medium up to 72 h when compared to controls. Dilution of the culture medium affected PC production, increasing it in 1/2 MS and especially in 1/4 MS. Cd uptake did not increase in the same way, but still hyperaccumulation levels were exceeded in all Cd treatments. BSO addition increased the sensitivity of D. viscosa to Cd and diminished Cd accumulation. Nevertheless, a poor correlation between PCs and Cd accumulation capacity was observed since the highest Cd content did not correspond to the highest PC levels. All these results obtained suggest that PCs are important in Cd accumulation and detoxification in D. viscosa and also that other mechanisms might be involved in these traits.

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

    Directory of Open Access Journals (Sweden)

    Kyo Shirai

    2012-01-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2000-06-30

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

  7. Development of optimal medium content for bioelements accumulation in Bacopa monnieri (L.) in vitro culture.

    Science.gov (United States)

    Łojewski, Maciej; Muszyńska, Bożena; Smalec, Agata; Reczyński, Witold; Opoka, Włodzimierz; Sułkowska-Ziaja, Katarzyna

    2014-10-01

    Bacopa monnieri is one of the most interesting plants from the Ayurveda system. The aims of present research were, basing on in vitro shoot culture of B. monnieri, to determine content and to evaluate the influence of physiologically important metabolites on the selected bioelements accumulation in biomass. The most significant increase in biomass production was observed in the culture medium enriched with 0.5 mg/L of anthranilic acid. In this medium also, the highest accumulation of Mg was noted. The highest concentration of iron was determined in B. monnieri in vitro culture enriched with 0.25 g/L of serine. The addition of L-tryptophan, magnesium sulfate, and zinc hydroaspartate caused only a small increase in the accumulation of copper in B. monnieri. Increase in Zn accumulation was obtained in biomass from in vitro culture of B. monnieri with the addition of magnesium sulfate and zinc hydroaspartate. In the case of Na, the maximum level of this element was in biomass from medium enriched with zinc hydroaspartate. Twofold increase in K concentration was obtained in biomass from cultures on medium with addition of serine and magnesium sulfate. The concentrations of Ca in biomass of all studied media were at the similar level.

  8. Cross-cultural and cross-ecotype production of a killer whale `excitement' call suggests universality

    Science.gov (United States)

    Rehn, Nicola; Filatova, Olga A.; Durban, John W.; Foote, Andrew D.

    2011-01-01

    Facial and vocal expressions of emotion have been found in a number of social mammal species and are thought to have evolved to aid social communication. There has been much debate about whether such signals are culturally inherited or are truly biologically innate. Evidence for the innateness of such signals can come from cross-cultural studies. Previous studies have identified a vocalisation (the V4 or `excitement' call) associated with high arousal behaviours in a population of killer whales in British Columbia, Canada. In this study, we compared recordings from three different socially and reproductively isolated ecotypes of killer whales, including five vocal clans of one ecotype, each clan having discrete culturally transmitted vocal traditions. The V4 call was found in recordings of each ecotype and each vocal clan. Nine independent observers reproduced our classification of the V4 call from each population with high inter-observer agreement. Our results suggest the V4 call may be universal in Pacific killer whale populations and that transmission of this call is independent of cultural tradition or ecotype. We argue that such universality is more consistent with an innate vocalisation than one acquired through social learning and may be linked to its apparent function of motivational expression.

  9. Reconstitution of Torso signaling in cultured cells suggests a role for both Trunk and Torso-like in receptor activation.

    Science.gov (United States)

    Amarnath, Smita; Stevens, Leslie M; Stein, David S

    2017-02-15

    Formation of the Drosophila embryonic termini is controlled by the localized activation of the receptor tyrosine kinase Torso. Both Torso and Torso's presumed ligand, Trunk, are expressed uniformly in the early embryo. Polar activation of Torso requires Torso-like, which is expressed by follicle cells adjacent to the ends of the developing oocyte. We find that Torso expressed at high levels in cultured Drosophila cells is activated by individual application of Trunk, Torso-like or another known Torso ligand, Prothoracicotropic Hormone. In addition to assays of downstream signaling activity, Torso dimerization was detected using bimolecular fluorescence complementation. Trunk and Torso-like were active when co-transfected with Torso and when presented to Torso-expressing cells in conditioned medium. Trunk and Torso-like were also taken up from conditioned medium specifically by cells expressing Torso. At low levels of Torso, similar to those present in the embryo, Trunk and Torso-like alone were ineffective but acted synergistically to stimulate Torso signaling. Our results suggest that Torso interacts with both Trunk and Torso-like, which cooperate to mediate dimerization and activation of Torso at the ends of the Drosophila embryo. © 2017. Published by The Company of Biologists Ltd.

  10. Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells.

    Science.gov (United States)

    Martin, G R

    1981-12-01

    This report describes the establishment directly from normal preimplantation mouse embryos of a cell line that forms teratocarcinomas when injected into mice. The pluripotency of these embryonic stem cells was demonstrated conclusively by the observation that subclonal cultures, derived from isolated single cells, can differentiate into a wide variety of cell types. Such embryonic stem cells were isolated from inner cell masses of late blastocysts cultured in medium conditioned by an established teratocarcinoma stem cell line. This suggests that such conditioned medium might contain a growth factor that stimulates the proliferation or inhibits the differentiation of normal pluripotent embryonic cells, or both. This method of obtaining embryonic stem cells makes feasible the isolation of pluripotent cells lines from various types of noninbred embryo, including those carrying mutant genes. The availability of such cell lines should made possible new approaches to the study of early mammalian development.

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

    Directory of Open Access Journals (Sweden)

    Céline Bouillon

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

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

    Directory of Open Access Journals (Sweden)

    Dorothea Alexander

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

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

    Science.gov (United States)

    Alexander, Dorothea; Rieger, Melanie; Klein, Christian; Ardjomandi, Nina; Reinert, Siegmar

    2013-01-01

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

  14. Effects of Proteins from Culture Medium on Surface Property of Silanes- Functionalized Magnetic Nanoparticles

    Directory of Open Access Journals (Sweden)

    Chen ZP

    2008-01-01

    Full Text Available Abstract Monodisperse magnetic nanoparticles (MNPs were synthesized by thermal decomposition of iron-oleate and functionalized with silanes bearing various functional groups such as amino group (NH2, short-chain poly(ethylene glycol (PEG, and carboxylic group (COOH. Then, silanes-functionalized magnetic nanoparticles (silanes-MNPs were incubated in cell culture medium plus fetal calf serum to investigate the effects of proteins from culture medium on surface property of MNPs. Zeta potential measurements showed that although surface charges of silanes-MNPs were different, they exhibited negative charges at neutral pH and approximate isoelectric points after they were incubated in cell culture medium. The reason was that silanes-MNPs could easily adsorb proteins from culture medium via non-covalent binding, resulting in the formation of protein-silanes-MNPs conjugates. Moreover, silanes-MNPs with various functional groups had different adsorption capacity to proteins, as confirmed by Coomassie blue fast staining method. The in vitro cell experiments showed that protein-silanes-MNPs had higher cellular uptake by cancer cells than silanes-MNPs.

  15. Uniform stable-isotope labeling in mammalian cells: formulation of a cost-effective culture medium

    NARCIS (Netherlands)

    Egorova-Zachernyuk, T.A.; Bosman, G.J.C.G.M.; Grip, W.J. de

    2011-01-01

    Uniform stable-isotope labeling of mammalian cells is achieved via a novel formulation of a serum-free cell culture medium that is based on stable-isotope-labeled autolysates and lipid extracts of various microbiological origin. Yeast autolysates allow complete replacement of individual amino acids

  16. Uniform stable-isotope labeling in mammalian cells: formulation of a cost-effective culture medium

    NARCIS (Netherlands)

    Egorova-Zachernyuk, T.A.; Bosman, G.J.C.G.M.; Grip, W.J. de

    2011-01-01

    Uniform stable-isotope labeling of mammalian cells is achieved via a novel formulation of a serum-free cell culture medium that is based on stable-isotope-labeled autolysates and lipid extracts of various microbiological origin. Yeast autolysates allow complete replacement of individual amino acids

  17. Effect of culture medium on biocalcification by Pseudomonas Putida, Lysinibacillus Sphaericus and Bacillus Subtilis

    Directory of Open Access Journals (Sweden)

    Márcia Aiko Shirakawa

    2011-06-01

    Full Text Available The objective of this study is to investigate the efficiency of calcium carbonate bioprecipitation by Lysinibacillus sphaericus, Bacillus subtilis and Pseudomonas putida, obtained from the Coleção de Culturas do Instituto Nacional de Controle de Qualidade em Saúde (INCQS, as a first step in determining their potential to protect building materials against water uptake. Two culture media were studied: modified B4 containing calcium acetate and 295 with calcium chloride. Calcium consumption in the two media after incubation with and without the bacterial inoculum was determined by atomic absorption analysis. Modified B4 gave the best results and in this medium Pseudomonas putida INQCS 113 produced the highest calcium carbonate precipitation, followed by Lysinibacillus sphaericus INQCS 414; the lowest precipitation was produced by Bacillus subtilis INQCS 328. In this culture medium XRD analysis showed that Pseudomonas putida and Bacillus subtilis precipitated calcite and vaterite polymorphs while Lysinibacillus sphaericus produced only vaterite. The shape and size of the crystals were affected by culture medium, bacterial strain and culture conditions, static or shaken. In conclusion, of the three strains Pseudomonas putida INQCS 113 in modified B4 medium gave the best results precipitating 96% of the calcium, this strain thus has good potential for use on building materials.

  18. Effect of culture medium on biocalcification by Pseudomonas Putida, Lysinibacillus Sphaericus and Bacillus Subtilis.

    Science.gov (United States)

    Shirakawa, Márcia Aiko; Cincotto, Maria Alba; Atencio, Daniel; Gaylarde, Christine C; John, Vanderley M

    2011-04-01

    The objective of this study is to investigate the efficiency of calcium carbonate bioprecipitation by Lysinibacillus sphaericus, Bacillus subtilis and Pseudomonas putida, obtained from the Coleção de Culturas do Instituto Nacional de Controle de Qualidade em Saúde (INCQS), as a first step in determining their potential to protect building materials against water uptake. Two culture media were studied: modified B4 containing calcium acetate and 295 with calcium chloride. Calcium consumption in the two media after incubation with and without the bacterial inoculum was determined by atomic absorption analysis. Modified B4 gave the best results and in this medium Pseudomonas putida INQCS 113 produced the highest calcium carbonate precipitation, followed by Lysinibacillus sphaericus INQCS 414; the lowest precipitation was produced by Bacillus subtilis INQCS 328. In this culture medium XRD analysis showed that Pseudomonas putida and Bacillus subtilis precipitated calcite and vaterite polymorphs while Lysinibacillus sphaericus produced only vaterite. The shape and size of the crystals were affected by culture medium, bacterial strain and culture conditions, static or shaken. In conclusion, of the three strains Pseudomonas putida INQCS 113 in modified B4 medium gave the best results precipitating 96% of the calcium, this strain thus has good potential for use on building materials.

  19. Biotransformation of tryptophan by liquid medium culture of Psilocybe coprophila (Basidiomycetes).

    Science.gov (United States)

    Alarcón, Julio; Foncea, Leyla; Aguila, Sergio; Alderete, Joel B

    2006-01-01

    Chemical reactions performed by fungi have been used as a modern tool in chemistry. In this work, we show the tryptophan biotransformation with Psilocybe coprophila on liquid culture medium. The results prove once more the versatility of fungi in performing a wide range of industrially attractive chemical reactions.

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

    Science.gov (United States)

    Kim, Jeongyeon; Tatar, Bradley; Choi, Jinsook

    2014-01-01

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

  1. Effect of culture medium on biocalcification by Pseudomonas Putida, Lysinibacillus Sphaericus and Bacillus Subtilis

    Science.gov (United States)

    Shirakawa, Márcia Aiko; Cincotto, Maria Alba; Atencio, Daniel; Gaylarde, Christine C.; John, Vanderley M.

    2011-01-01

    The objective of this study is to investigate the efficiency of calcium carbonate bioprecipitation by Lysinibacillus sphaericus, Bacillus subtilis and Pseudomonas putida, obtained from the Coleção de Culturas do Instituto Nacional de Controle de Qualidade em Saúde (INCQS), as a first step in determining their potential to protect building materials against water uptake. Two culture media were studied: modified B4 containing calcium acetate and 295 with calcium chloride. Calcium consumption in the two media after incubation with and without the bacterial inoculum was determined by atomic absorption analysis. Modified B4 gave the best results and in this medium Pseudomonas putida INQCS 113 produced the highest calcium carbonate precipitation, followed by Lysinibacillus sphaericus INQCS 414; the lowest precipitation was produced by Bacillus subtilis INQCS 328. In this culture medium XRD analysis showed that Pseudomonas putida and Bacillus subtilis precipitated calcite and vaterite polymorphs while Lysinibacillus sphaericus produced only vaterite. The shape and size of the crystals were affected by culture medium, bacterial strain and culture conditions, static or shaken. In conclusion, of the three strains Pseudomonas putida INQCS 113 in modified B4 medium gave the best results precipitating 96% of the calcium, this strain thus has good potential for use on building materials. PMID:24031661

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

    Science.gov (United States)

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

    2017-09-01

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

  3. Anglo-Saxon imperialism through cultural goods: titles suggested for young readers in Portugal

    Directory of Open Access Journals (Sweden)

    Maria João Ferro

    2017-01-01

    Full Text Available Translation plays a vital role in the current multilingual and multicultural society. It is a fundamental infrastructure of globalization, functioning as an essential tool for the circulation of information, knowledge, and culture. Although English can be considered a lingua franca in certain sectors of society, the role of translation has not lessened with its rise. In fact, translation is a strong indicator of the relationships established between countries: international translation flows depend on the (imbalances of power in the world. In this paper I analyse relevant data to ascertain the weight of works originally written in English and translated into Portuguese, resorting to the national reading plan (PNL. After briefly addressing the influence of geopolitics in the trade of cultural goods and describing the world system of translation, I analyse the list of 345 books suggested in the national reading plan for 12-15 year olds. Almost half of these books are translations; however, this does not imply that the list is sufficiently multicultural, quite on the contrary as it shows the dominance of books written in English. A lot more can be done to diversify the source languages and expose Portuguese young readers to a wider variety of cultures.

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

    Directory of Open Access Journals (Sweden)

    Williams Sylvain

    2006-03-01

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

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

    Science.gov (United States)

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

    2016-01-01

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

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

    Science.gov (United States)

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

    2015-12-01

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

  7. Optimization of culture medium for the continuous cultivation of the microalga Haematococcus pluvialis.

    Science.gov (United States)

    Fábregas, J; Domínguez, A; Regueiro, M; Maseda, A; Otero, A

    2000-05-01

    The freshwater microalga Haematococcus pluvialis is one of the best microbial sources of the carotenoid astaxanthin, but this microalga shows low growth rates and low final cell densities when cultured with traditional media. A single-variable optimization strategy was applied to 18 components of the culture media in order to maximize the productivity of vegetative cells of H. pluvialis in semicontinuous culture. The steady-state cell density obtained with the optimized culture medium at a daily volume exchange of 20% was 3.77 x 10(5) cells ml(-1), three times higher than the cell density obtained with Bold basal medium and with the initial formulation. The formulation of the optimal Haematococcus medium (OHM) is (in g l(-1)) KNO3 0.41, Na2HPO4 0.03, MgSO4 x 7H2O 0.246, CaCl2 x 2H2O 0.11, (in mg l(-1)) Fe(III)citrate x H2O 2.62, CoCl2 x 6H2O 0.011, CuSO4 x 5H2O 0.012, Cr2O3 0.075, MnCl2 x 4H2O 0.98, Na2MoO4 x 2H2O 0.12, SeO2 0.005 and (in microg l(-1)]) biotin 25, thiamine 17.5 and B12 15. Vanadium, iodine, boron and zinc were demonstrated to be non-essential for the growth of H. pluvialis. Higher steady-state cell densities were obtained by a three-fold increase of all nutrient concentrations but a high nitrate concentration remained in the culture medium under such conditions. The high cell productivities obtained with the new optimized medium can serve as a basis for the development of a two-stage technology for the production of astaxanthin from H. pluvialis.

  8. Emulsion culture: a miniaturized library screening system based on micro-droplets in an emulsified medium.

    Science.gov (United States)

    Kojima, Takaaki; Nagao, Nobuhito; Ando, Daisuke; Ojima, Teruyo; Kawarasaki, Yasuaki; Kobayashi, Isao; Nakajima, Mitsutoshi; Nakano, Hideo

    2011-09-01

    A typical library screen in directed evolution primarily requires physical separation of the clones on agar plates followed by detection of clones with improved properties; using this method only limited numbers of clones relative to the number of potential variations can be assessed. In particular, screening for a secretory enzyme is difficult to perform at high clone density, because of diffusion of the signal or unfavorable utilization of the reaction product by neighboring clones. In this study, we have developed a novel method of enrichment culture: "Emulsion Culture", i.e., segregated replication of clones in an emulsified culture medium. Clones expressing enzyme-variants are separately distributed to small (up to 50 μm in diameter), segregated compartments composed of a droplet of medium to form several tens of millions of microcolonies in a milliliter of medium, which allows a miniaturized, in-bulk screening of clones. We applied this culture method to yeast clones expressing secretory beta-galactosidase to analyze the enrichment factor achieved. A high-density screen for a signal peptide sequence that maximizes extracellular production of the enzyme was also performed to demonstrate the practicability of this culture method. In addition, micro-channel emulsification was tested as a method of forming uniformly-sized compartments in the emulsion.

  9. Inorganic ions in the medium modify tropane alkaloids and riboflavin output in Hyoscyamus niger root cultures

    Directory of Open Access Journals (Sweden)

    Katrin Pudersell

    2012-01-01

    Full Text Available Context: Hyoscyamus niger L. (Solanaceae roots are rich of tropane alkaloids, such as hyoscyamine and scopolamine are used as the source of raw material for the pharmaceutical industry. Aims: The aim of the present study was to investigate the effect of calcium, magnesium, and iron ions on the production of tropane alkaloids and excretion of riboflavin in H. niger root cultures. Materials and Methods: The calcium, magnesium, or iron enriched/deprived Murashige and Skoog (MS growth medium were used for culture of H. niger root tissues. The secondary metabolites were quantified using high performance liquid chromatography with ultraviolet detector (HPLC-UV and fluorimetry techniques. Results: An increased calcium content in the medium unidirectionally reduced hyoscyamine, while increasing scopolamine production with only a moderate impact on riboflavin excretion. Manipulations with magnesium and iron contents in the medium resulted in divergent changes in hyoscyamine, scopolamine, and riboflavin concentrations. Conclusions: Our results show that increased calcium ion content in the Murashige and Skoog medium may be used for the intensification of the scopolamine production in H. niger root cultures.

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

    Science.gov (United States)

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

    2015-01-01

    Fewer studies have assessed the outdoor cultivation of Spirulina maxima compared with S. platensis, although the protein content of S. maxima is higher than S. platensis. Spirulina growth medium requires an increased amount of NaHCO3, Na2CO3, and NaNO3, which increases the production cost. Therefore, the current study used a low-cost but high-efficiency biomass production medium (Medium M-19) after testing 33 different media. The medium depth of 25 cm (group A) was sub-divided into A1 (50% cover with a black curtain (PolyMax, 12 oz ultra-blackout), A2 (25% cover), and A3 (no cover). Similarly the medium depths of 30 and 35 cm were categorized as groups B (B1, B2, and B3) and C (C1, C2, and C3), respectively, and the effects of depth and surface light availability on growth and biomass production were assessed. The highest biomass production was 2.05 g L-1 in group A2, which was significantly higher (p < 0.05) than that in all other groups and sub-groups. Spirulina maxima died in B1 and C1 on the fifth day of culture. The biochemical composition of the biomass obtained from A2 cultures, including protein, carbohydrate, lipid, moisture, and ash, was 56.59%, 14.42%, 0.94%, 5.03%, and 23.02%, respectively. Therefore, S. maxima could be grown outdoors with the highest efficiency in urea-enriched medium at a 25-cm medium depth with 25% surface cover or uncovered. PMID:26691456

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

    Directory of Open Access Journals (Sweden)

    Md-Abu Affan

    2015-01-01

    Full Text Available Fewer studies have assessed the outdoor cultivation of Spirulina maxima compared with S. platensis, although the protein content of S. maxima is higher than S. platensis. Spirulina growth medium requires an increased amount of NaHCO3, Na2CO3, and NaNO3, which increases the production cost. Therefore, the current study used a low-cost but high-efficiency biomass production medium (Medium M-19 after testing 33 different media. The medium depth of 25 cm (group A was sub-divided into A1 (50% cover with a black curtain (PolyMax, 12 oz ultra-blackout, A2 (25% cover, and A3 (no cover. Similarly the medium depths of 30 and 35 cm were categorized as groups B (B1, B2, and B3 and C (C1, C2, and C3, respectively, and the effects of depth and surface light availability on growth and biomass production were assessed. The highest biomass production was 2.05 g L-1 in group A2, which was significantly higher (p < 0.05 than that in all other groups and sub-groups. Spirulina maxima died in B1 and C1 on the fifth day of culture. The biochemical composition of the biomass obtained from A2 cultures, including protein, carbohydrate, lipid, moisture, and ash, was 56.59%, 14.42%, 0.94%, 5.03%, and 23.02%, respectively. Therefore, S. maxima could be grown outdoors with the highest efficiency in urea-enriched medium at a 25-cm medium depth with 25% surface cover or uncovered.

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

    Science.gov (United States)

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

    2015-01-01

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

  13. 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.

    Science.gov (United States)

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

    2005-06-25

    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.

  14. [Use of transport medium in sputum bacterial culture examination of lower airway infection].

    Science.gov (United States)

    Muraki, Masato; Kitaguchi, Sayako; Ichihashi, Hideo; Tsuji, Fumio; Ohmori, Takashi; Haraguchi, Ryuta; Tohda, Yuji

    2006-06-01

    Our medical institution does not have a bacterial culture facility, requiring outsourcing of bacterial culture tests. Due to the time elapsed from the time of specimen collection to culturing, the identification of causative bacteria in respiratory tract infections tends to be difficult. We therefore used transport medium for sputum bacteria examinations. Expectorated purulent or purulent-mucous sputum specimens were collected from 32 patients with lower respiratory tract infection. We divided each of the sputum specimens into the two treatment groups: transport medium (Seedswab gamma2) ndar and stad disinfection container. Paired samples prepared from each patient were sent out for bacterial culture together. The time elapsed from collection to delivery to the lab were as follows: day 0 (same day, n = 14 patients), day 1 (n = 15), day 2 (n = 2), and day 3 (n = 1). The identified causative bacteria were Streptococcus pneumoniae (n = 6 patients), Haemophilus influenzae (n =5), Pseudomonas aeruginosa (n = 4), Staphylococcus aureus (n = 2), Moraxella catarrhalis (n = 2), Klebsiella pneumoniae (n = 1), and Streptococcus agalactiae (n = 1). Samples prepared by each of the two methods gave similar results. The utility of transport medium for examination of general bacteria for lower airway infection from sputum samples was not demonstrated. The rate of detection of bacteria decreased, when the transport of samples was delayed. Therefore, we need to send the sputum specimens as quickly as possible.

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

    Directory of Open Access Journals (Sweden)

    Carlos Andrés Zárate-Chaves

    2013-01-01

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

  16. Nuclear and mitochondrial DNA in blastocoele fluid and embryo culture medium: evidence and potential clinical use.

    Science.gov (United States)

    Hammond, Elizabeth R; Shelling, Andrew N; Cree, Lynsey M

    2016-08-01

    The ability to screen embryos for aneuploidy or inherited disorders in a minimally invasive manner may represent a major advancement for the future of embryo viability assessment. Recent studies have demonstrated that both blastocoele fluid and embryo culture medium contain genetic material, which can be isolated and subjected to downstream genetic analysis. The blastocoele fluid may represent an alternative source of nuclear DNA for aneuploidy testing, although the degree to which the isolated genetic material is solely representative of the developing embryo is currently unclear. In addition to nuclear DNA, mitochondrial DNA (mtDNA) can be detected in the embryo culture medium. Currently, the origin of this nuclear and mtDNA has not been fully evaluated and there are several potential sources of contamination that may contribute to the genetic material detected in the culture medium. There is however evidence that the mtDNA content of the culture medium is related to embryo fragmentation levels and its presence is predictive of blastulation, indicating that embryo development may influence the levels of genetic material detected. If the levels of genetic material are strongly related to aspects of embryo quality, then this may be a novel biomarker of embryo viability. If the genetic material does have an embryo origin, the mechanisms by which DNA may be released into the blastocoele fluid and embryo culture medium are unknown, although apoptosis may play a role. While the presence of this genetic material is an exciting discovery, the DNA in the blastocoele fluid and embryo culture medium appears to be of low yield and integrity, which makes it challenging to study. Further research aimed at assessing the methodologies used for both isolating and analysing this genetic material, as well as tracing its origin, are needed in order to evaluate its potential for clinical use. Should such methodologies prove to be routinely successful and the DNA recovered

  17. Evaluation of new transport medium for detection of herpes simplex virus by culture and direct enzyme-linked immunosorbent assay.

    Science.gov (United States)

    Ogburn, J R; Hoffpauir, J T; Cole, E; Hood, K; Michael, D; Nguyen, T; Raden, S; Raju, B; Reisinger, V; Oefinger, P E

    1994-12-01

    The transport medium Multi-Microbe Media (M4) was evaluated prospectively by culture and direct enzyme-linked immunosorbent assay (ELISA) for detection of herpes simplex virus from 473 specimens. In addition, 377 specimens in Bartels Viral Transport Medium were evaluated. By using culture as a "gold standard," the ELISA sensitivity was approximately 85%, while the specificities exceeded 96% for both media.

  18. Feeder-free culture of human embryonic stem cells in conditioned medium for efficient genetic modification.

    Science.gov (United States)

    Braam, Stefan R; Denning, Chris; Matsa, Elena; Young, Lorraine E; Passier, Robert; Mummery, Christine L

    2008-01-01

    Realizing the potential of human embryonic stem cells (hESCs) in research and commercial applications requires generic protocols for culture, expansion and genetic modification that function between multiple lines. Here we describe a feeder-free hESC culture protocol that was tested in 13 independent hESC lines derived in five different laboratories. The procedure is based on Matrigel adaptation in mouse embryonic fibroblast conditioned medium (CM) followed by monolayer culture of hESC. When combined, these techniques provide a robust hESC culture platform, suitable for high-efficiency genetic modification via plasmid transfection (using lipofection or electroporation), siRNA knockdown and viral transduction. In contrast to other available protocols, it does not require optimization for individual lines. hESC transiently expressing ectopic genes are obtained within 9 d and stable transgenic lines within 3 weeks.

  19. Best practices for the use and evaluation of animal serum as a component of cell culture medium.

    Science.gov (United States)

    Nims, Raymond W; Harbell, John W

    2017-07-21

    Animal serum is a common additive for cell culture medium and is often required at 5 to 10% (v/v) for the attachment and growth of primary and continuous anchorage-dependent (monolayer) cultures. The use of animal serum in cell culture medium confers several advantages and also some risks. This article discusses the use of animal serum as a component of cell culture medium. The best practices associated with the sourcing, storage, thawing, testing, and mitigation of risk associated with the use of animal sera are among the topics described in this article.

  20. Culturing Selenastrum capricornutum (Chlorophyta) in a synthetic algal nutrient medium with defined mineral particulates

    Science.gov (United States)

    Kuwabara, J.S.; Davis, J.A.; Chang, Cecily C.Y.

    1985-01-01

    Algal nutrient studies in chemically-defined media typically employ a synthetic chelator to prevent iron hydroxide precipitation. Micronutrient-particulate interactions may, however, significantly affect chemical speciation and hence biovailability of these nutrients in natural waters. A technique is described by which Selenastrum capricornutum Printz (Chlorophyta) may be cultured in a medium where trace metal speciation (except iron) is controlled, not by organic chelation, but by sorption onto titanium dioxide. Application of this culturing protocol in conjunction with results from sorption studies of nutrient ions on mineral particles provides a means of studying biological impacts of sorptive processes in aquatic environments. ?? 1985 Dr W. Junk Publishers.

  1. Development of an animal-component free medium for vero cells culture.

    Science.gov (United States)

    Rourou, Samia; van der Ark, Arno; van der Velden, Tiny; Kallel, Héla

    2009-01-01

    This work describes the development of an animal-component free medium (IPT-AFM) that allows an optimal growth of Vero cells, an adherent cell line used for the production of viral vaccines. Statistical experimental design was applied to identify crucial nutrients that affect cell growth. Using Medium 199 or MEM as a basal medium, a serum-free medium (SFM) referred as IPT-SFM that only enclosed transferrin as a component of animal origin was developed at first. Then, the composition of IPT-SFM was further improved to obtain an animal-component free medium named IPT-AFM. IPT-AFM contains M199 as a basal medium, plant hydrolysates, epidermal growth factor, ethanolamine, ferric citrate, and vitamin C. Among various plant hydrolysates, specific combinations of soy (Hypep 1510) and wheat gluten (Hypeps 4601 and 4605) hydrolysates, were identified to promote cell growth; whereas individual Hypeps had a minor positive effect on cell growth. Nevertheless, the removal of serum did influence cell attachment. Coating tissue-culture flasks with teleostean, a product extracted from cold water fish skin, had not only enhanced cell attachment but also improved cell growth performance in static cultures. Different non-animal proteases were also assessed as an alternative to trypsin. TrypLE Select, a recombinant trypsin, gave the best cell growth performances. Kinetics of cell growth in IPT-AFM were investigated in T-flasks, cell growth was comparable with that obtained in MEM+10% fetal calf serum (FCS). A mean cell division number equal to 2.26 +/- 0.18 and a specific growth rate micro 0.019 +/- 0.003 h(-1) were achieved in IPT-AFM. (c) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009.

  2. PROPAGATION OF Portulaca oleracea L. IN LIQUID MEDIUM: IMPLICATIONS OF PLANT GROWTH REGULATORS IN CULTURE

    Directory of Open Access Journals (Sweden)

    Mahipal S. Shekhawat

    2015-02-01

    Full Text Available Portulaca oleracea L. is a medicinal plant, growing in warm and moist regions of north hemisphere of the world. A protocol for in vitro propagation using nodal shoot segments as explants has been outlined. The percent shoot response with shoot induction rate, 6.4 ± 0.7 shoots per explant, was achieved when cultured on agar-gelled Murashige and Skoog (MS medium containing 2.0 mg/L of BAP (6-benzylaminopurine. The cultures were amplified by passages on MS medium with 1.0 mg/L each BAP and kinetin (Kn. The best shoot amplification (37.5±0.9 shoots per vessel was achieved by subculturing of in vitro regenerated shoot clumps on liquid MS medium. Shoots regenerated in vitro by both the processes were rooted on ½ strength of MS medium + 2.0 mg/L of indole-3 butyric acid (IBA. Ninety six percent of the shoots rooted in vitro. The in vitro rooted plantlets were hardened under different regimes of temperature and humidity in the greenhouse. The hardened plantlets were transferred to mixture of soil and manure in polybags.

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

    Directory of Open Access Journals (Sweden)

    Pusztai Beáta

    2015-08-01

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

  4. Mutagenesis breeding of high echinocandin B producing strain and further titer improvement with culture medium optimization.

    Science.gov (United States)

    Zou, Shu-Ping; Zhong, Wei; Xia, Chao-Jie; Gu, Ya-Nan; Niu, Kun; Zheng, Yu-Guo; Shen, Yin-Chu

    2015-10-01

    A combination of microbial strain improvement and statistical optimization is investigated to maximize echinocandin B (ECB) production from Aspergillus nidulans ZJB-0817. A classical sequential mutagenesis was studied first by using physical (ultraviolet irradiation at 254 nm) and chemical mutagens (lithium chloride and sodium nitrite). Mutant strain ULN-59 exhibited 2.1-fold increase in ECB production to 1583.1 ± 40.9 mg/L when compared with the parent strain (750.8 ± 32.0 mg/L). This is the first report where mutagenesis is applied in Aspergillus to improve ECB production. Further, fractional factorial design and central composite design were adopted to optimize the culture medium for increasing ECB production by the mutant ULN-59. Results indicated that four culture media including peptone, K2HPO4, mannitol and L-ornithine had significant effects on ECB production. The optimized medium provided another 1.4-fold increase in final ECB concentration to 2285.6 ± 35.6 mg/L compared to the original medium. The results of this study indicated the combined application of a classical mutation and medium optimization can improve effectively ECB production from A. nidulans and could be a promising tool to improve other secondary metabolites production by fungal strains.

  5. Supplementation of insulin-transferrin-selenium to embryo culture medium improves the in vitro development of pig embryos.

    Science.gov (United States)

    Das, Ziban Chandra; Gupta, Mukesh Kumar; Uhm, Sang Jun; Lee, Hoon Taek

    2014-08-01

    Insulin, transferrin and selenium (ITS) supplementation to oocyte maturation medium improves the post-fertilization embryonic development in pigs. ITS is also commonly used as a supplement for the in vitro culture (IVC) of embryos and stem cells in several mammalian species. However, its use during IVC of pig embryos has not been explored. This study investigated the effect of ITS supplementation to IVC medium on the in vitro development ability of pig embryos produced by parthenogenetic activation (PA), in vitro fertilization (IVF) or somatic cell nuclear transfer (SCNT). We observed that ITS had no significant effect on the rate of first cleavage (P > 0.05). However, the rate of blastocyst formation in ITS-treated PA (45.3 ± 1.9 versus 27.1 ± 2.3%), IVF (31.6 ± 0.6 versus 23.5 ± 0.6%) and SCNT (17.6 ± 2.3 versus 10.7 ± 1.4%) embryos was significantly higher (P Culture of PA embryos in the presence of ITS also enhanced the expansion and hatching ability (29.1 ± 3.0 versus 18.2 ± 3.8%; P 0.05). Taken together, these data suggest that supplementation of ITS to the IVC medium exerts a beneficial but differential effect on pig embryos that varies with the method of embryo production in vitro.

  6. Effects of bovine serum proteins in culture medium on post-warming survival of bovine blastocysts developed in vitro.

    Science.gov (United States)

    Ohboshi, S; Etoh, T; Sakamoto, K; Fujihara, N; Yoshida, T; Tomogane, H

    1997-04-15

    Experiments were conducted to investigate the factors affecting the survival of bovine blastocysts produced in vitro after cryopreservation by vitrification. Zygotes were obtained by in vitro maturation and fertilization of oocytes. Embryos used in this study were developed in vitro at Day 7 and 8 (Day 0 = insemination day) in modified synthetic oviduct fluid medium supplemented with calf serum or BSA. Embryos were cryopreserved in a two-step protocol consisting of exposure to 10% ethylene glycol for 5 min, followed by the original vitrification solution (designated as VS) consisting of 40% (v/v) ethylene glycol, 6% (w/v) polyethylene glycol and 0.5 M sucrose in phosphate-buffered saline for 1 min. After warming, embryos were cultured in modified TCM-199 for an in vitro survival assay. The highest survival rate was obtained from the warmed embryos developed at Day 7 in medium supplemented with BSA (82.6%), and there were significant differences between results with calf scrum and BSA treatment (42.4 and 70.7%, respectively; P < 0.01). However, there were no significant differences in the cell numbers of embryos among the treatments. These results suggest that the survival of embryos developed in medium with BSA is superior to that of embryos developed in medium containing calf serum, although the cell numbers of the embryos developed under both media were similar.

  7. Development and validation of a liquid medium (M7H9C) for routine culture of Mycobacterium avium subsp. paratuberculosis to replace modified Bactec 12B medium.

    Science.gov (United States)

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

    2013-12-01

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

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

    Science.gov (United States)

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

    2015-05-01

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

  9. Culturing and direct PCR suggest prevalent host generalism among diverse fungal endophytes of tropical forest grasses.

    Science.gov (United States)

    Higgins, K Lindsay; Coley, Phyllis D; Kursar, Thomas A; Arnold, A Elizabeth

    2011-01-01

    Most studies examining endophytic fungi associated with grasses (Poaceae) have focused on agronomically important species in managed ecosystems or on wild grasses in subtropical, temperate and boreal grasslands. However grasses first arose in tropical forests, where they remain a significant and diverse component of understory and forest-edge communities. To provide a broader context for understanding grass-endophyte associations we characterized fungal endophyte communities inhabiting foliage of 11 species of phylogenetically diverse C(3) grasses in the understory of a lowland tropical forest at Barro Colorado Island, Panama. Our sample included members of early-arising subfamilies of Poaceae that are endemic to forests, as well as more recently arising subfamilies that transitioned to open environments. Isolation on culture media and direct PCR and cloning revealed that these grasses harbor species-rich and phylogenetically diverse communities that lack the endophytic Clavicipitaceae known from diverse woodland and pasture grasses in the temperate zone. Both the incidence and diversity of endophytes was consistent among grass species regardless of subfamily, clade affiliation or ancestral habitat use. Genotype and phylogenetic analyses suggest that these endophytic fungi are predominantly host generalists, shared not only among distinctive lineages of Poaceae but also with non-grass plants at the same site.

  10. Aquatic flower-inspired cell culture platform with simplified medium exchange process for facilitating cell-surface interaction studies.

    Science.gov (United States)

    Hong, Hyeonjun; Park, Sung Jea; Han, Seon Jin; Lim, Jiwon; Kim, Dong Sung

    2016-02-01

    Establishing fundamentals for regulating cell behavior with engineered physical environments, such as topography and stiffness, requires a large number of cell culture experiments. However, cell culture experiments in cell-surface interaction studies are generally labor-intensive and time-consuming due to many experimental tasks, such as multiple fabrication processes in sample preparation and repetitive medium exchange in cell culture. In this work, a novel aquatic flower-inspired cell culture platform (AFIP) is presented. AFIP aims to facilitate the experiments on the cell-surface interaction studies, especially the medium exchange process. AFIP was devised to capture and dispense cell culture medium based on interactions between an elastic polymer substrate and a liquid medium. Thus, the medium exchange can be performed easily and without the need of other instruments, such as a vacuum suction and pipette. An appropriate design window of AFIP, based on scaling analysis, was identified to provide a criterion for achieving stability in medium exchange as well as various surface characteristics of the petal substrates. The developed AFIP, with physically engineered petal substrates, was also verified to exchange medium reliably and repeatedly. A closed structure capturing the medium was sustained stably during cell culture experiments. NIH3T3 proliferation results also demonstrated that AFIP can be applied to the cell-surface interaction studies as an alternative to the conventional method.

  11. Prediction of embryo implantation potential by mass spectrometry fingerprinting of the culture medium.

    Science.gov (United States)

    Cortezzi, Sylvia Sanches; Cabral, Elaine Cristina; Trevisan, Marcello Garcia; Ferreira, Christina Ramires; Setti, Amanda Souza; Braga, Daniela Paes de Almeida Ferreira; Figueira, Rita de Cássia Sávio; Iaconelli, Assumpto; Eberlin, Marcos Nogueira; Borges, Edson

    2013-05-01

    This study has evaluated the performance of a multivariate statistical model to predict embryo implantation potential by processing data from the chemical fingerprinting of culture medium samples used for human embryo culture. The culture medium for 113 embryos from 55 patients undergoing ICSI was collected after embryo transfer. The samples were split into positive (n=29) and negative (n=84) implantation groups according their implantation outcomes (100% or 0% implantation). The samples were individually diluted and analyzed by electrospray ionization mass spectrometry (ESI-MS). The m/z ratios and relative abundances of the major ions in each spectrum were considered for partial least square discriminant analysis. Data were divided into two subsets (calibration and validation), and the models were evaluated and applied to the validation set. A total of 5987 ions were observed in the groups. The multivariate statistical model described more than 82% of the data variability. Samples of the positive group were correctly identified with 100% probability and negative samples with 70%. The culture media used for embryos that were positive or negative for successful implantation showed specific biochemical signatures that could be detected in a fast, simple, and noninvasive way by ESI-MS. To our knowledge, this is the first report that uses MS fingerprinting to predict human embryo implantation potential. This biochemical profile could help the selection of the most viable embryo, improving single-embryo transfer and thus eliminating the risk and undesirable outcomes of multiple pregnancies.

  12. Rhamnolipids elicit the same cytotoxic sensitivity between cancer cell and normal cell by reducing surface tension of culture medium.

    Science.gov (United States)

    Jiang, Lifang; Shen, Chong; Long, Xuwei; Zhang, Guoliang; Meng, Qin

    2014-12-01

    Biosurfactant rhamnolipids have been claimed to show biological activities of inhibiting the proliferation of cancer cells. In this study, the cytotoxicity of rhamnolipids was examined on four cancer cells (HepG2, Caco-2, Hela, MCF-7 cells) and two normal cells (HK-2 cell, primary hepatocyte). Interestingly, both cancer cells and normal cells exhibited similar sensitivities to the addition of rhamnolipids in culture medium, and the cytotoxicity was largely attenuated by the presence of fetal bovine serum (FBS) in culture medium. In correlation of the mono-/di-rhamnolipid cytotoxicity with the surface tension of culture medium, it was found that rhamnolipids triggered cytotoxicity whenever the surface tension of culture medium decreased below 41 mN/m irrespective of the FBS content in culture medium, cell line, or rhamnolipid congener. Similarly, each chemical surfactant (Tween-80, sodium dodecyl sulfate, and sodium dodecyl benzene sulfonate) could cause cytotoxicity on HepG2 cells whenever its addition made the surface tension under 41 mN/m in culture medium with or without the presence of FBS. It seems that rhamnolipids, like chemical surfactants, exhibited cytotoxicity by reducing the surface tension of culture medium rather than by changing its specific molecular structure, which had no selection on tumor cells. This study could offer helps to correct the misleading biological activity of rhamnolipids and to avoid the possible large wastes of time and expenses on developing the applications in antitumor drugs.

  13. Silver nanoparticles inhibit fish gill cell proliferation in protein-free culture medium.

    Science.gov (United States)

    Yue, Yang; Behra, Renata; Sigg, Laura; Schirmer, Kristin

    2016-10-01

    While short-term exposures of vertebrate cells, such as from fish, can be performed in defined, serum-free media, long-term cultures generally require addition of growth factors and proteins, normally supplied with a serum supplement. However, proteins are known to alter nanoparticle properties by binding to nanoparticles. Therefore, in order to be able to study nanoparticle-cell interactions for extended periods, the rainbow trout (Oncorhynchus mykiss) gill cell line, RTgill-W1, was adapted to proliferate in a commercial, serum-free medium, InVitrus VP-6. The newly adapted cell strain was named RTgill-W1-pf (protein free). These cells proliferate at a speed similar to the RTgill-W1 cells cultured in a fully supplemented medium containing 5% fetal bovine serum. As well, they were successfully cryopreserved in liquid nitrogen and fully recovered after thawing. Yet, senescence set in after about 10 passages in InVitrus VP-6 medium, revealing that this medium cannot fully support long-term culture of the RTgill-W1 strain. The RTgill-W1-pf cell line was subsequently applied to investigate the effect of silver nanoparticles (AgNP) on cell proliferation over a period of 12 days. Indeed, cell proliferation was inhibited by 10 μM AgNP. This effect correlated with high levels of silver being associated with the cells. The new cell line, RTgill-W1-pf, can serve as a unique representation of the gill cell-environment interface, offering novel opportunities to study nanoparticle-cell interactions without serum protein interference.

  14. High diversity and suggested endemicity of culturable Actinobacteria in an extremely oligotrophic desert oasis

    Directory of Open Access Journals (Sweden)

    Hector Fernando Arocha-Garza

    2017-05-01

    Full Text Available The phylum Actinobacteria constitutes one of the largest and anciently divergent phyla within the Bacteria domain. Actinobacterial diversity has been thoroughly researched in various environments due to its unique biotechnological potential. Such studies have focused mostly on soil communities, but more recently marine and extreme environments have also been explored, finding rare taxa and demonstrating dispersal limitation and biogeographic patterns for Streptomyces. To test the distribution of Actinobacteria populations on a small scale, we chose the extremely oligotrophic and biodiverse Cuatro Cienegas Basin (CCB, an endangered oasis in the Chihuahuan desert to assess the diversity and uniqueness of Actinobacteria in the Churince System with a culture-dependent approach over a period of three years, using nine selective media. The 16S rDNA of putative Actinobacteria were sequenced using both bacteria universal and phylum-specific primer pairs. Phylogenetic reconstructions were performed to analyze OTUs clustering and taxonomic identification of the isolates in an evolutionary context, using validated type species of Streptomyces from previously phylogenies as a reference. Rarefaction analysis for total Actinobacteria and for Streptomyces isolates were performed to estimate species’ richness in the intermediate lagoon (IL in the oligotrophic Churince system. A total of 350 morphologically and nutritionally diverse isolates were successfully cultured and characterized as members of the Phylum Actinobacteria. A total of 105 from the total isolates were successfully subcultured, processed for DNA extraction and 16S-rDNA sequenced. All strains belong to the order Actinomycetales, encompassing 11 genera of Actinobacteria; the genus Streptomyces was found to be the most abundant taxa in all the media tested throughout the 3-year sampling period. Phylogenetic analysis of our isolates and another 667 reference strains of the family Streptomycetaceae

  15. High diversity and suggested endemicity of culturable Actinobacteria in an extremely oligotrophic desert oasis.

    Science.gov (United States)

    Arocha-Garza, Hector Fernando; Canales-Del Castillo, Ricardo; Eguiarte, Luis E; Souza, Valeria; De la Torre-Zavala, Susana

    2017-01-01

    The phylum Actinobacteria constitutes one of the largest and anciently divergent phyla within the Bacteria domain. Actinobacterial diversity has been thoroughly researched in various environments due to its unique biotechnological potential. Such studies have focused mostly on soil communities, but more recently marine and extreme environments have also been explored, finding rare taxa and demonstrating dispersal limitation and biogeographic patterns for Streptomyces. To test the distribution of Actinobacteria populations on a small scale, we chose the extremely oligotrophic and biodiverse Cuatro Cienegas Basin (CCB), an endangered oasis in the Chihuahuan desert to assess the diversity and uniqueness of Actinobacteria in the Churince System with a culture-dependent approach over a period of three years, using nine selective media. The 16S rDNA of putative Actinobacteria were sequenced using both bacteria universal and phylum-specific primer pairs. Phylogenetic reconstructions were performed to analyze OTUs clustering and taxonomic identification of the isolates in an evolutionary context, using validated type species of Streptomyces from previously phylogenies as a reference. Rarefaction analysis for total Actinobacteria and for Streptomyces isolates were performed to estimate species' richness in the intermediate lagoon (IL) in the oligotrophic Churince system. A total of 350 morphologically and nutritionally diverse isolates were successfully cultured and characterized as members of the Phylum Actinobacteria. A total of 105 from the total isolates were successfully subcultured, processed for DNA extraction and 16S-rDNA sequenced. All strains belong to the order Actinomycetales, encompassing 11 genera of Actinobacteria; the genus Streptomyces was found to be the most abundant taxa in all the media tested throughout the 3-year sampling period. Phylogenetic analysis of our isolates and another 667 reference strains of the family Streptomycetaceae shows that our

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

    Science.gov (United States)

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

    2016-11-01

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

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

    NARCIS (Netherlands)

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

    2016-01-01

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

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

    NARCIS (Netherlands)

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

    2016-01-01

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

  19. Developmental features of rat cerebellar neural cells cultured in a chemically defined medium

    Energy Technology Data Exchange (ETDEWEB)

    Gallo, V.; Ciotti, M.T.; Aloisi, F.; Levi, G.

    1986-01-01

    We studied some aspects of the differentiation of rat cerebellar neural cells obtained from 8-day postnatal animals and cultured in a serum-free, chemically defined medium (CDM). The ability of the cells to take up radioactive transmitter amino acids was analyzed autoradiographically. The L-glutamate analogue /sup 3/H-D-aspartate was taken up by astroglial cells, but not by granule neurons, even in late cultures (20 days in vitro). This is in agreement with the lack of depolarization-induced release of /sup 3/H-D-aspartate previously observed in this type of culture. In contrast, /sup 3/H-(GABA) was scarcely accumulated by glial-fibrillary-acidic-protein (GFAP)-positive astrocytes, but taken up by glutamate-decarboxylase-positive inhibitory interneurons and was released in a Ca2+-dependent way upon depolarization: /sup 3/H-GABA evoked release progressively increased with time in culture. Interestingly, the expression of the vesicle-associated protein synapsin I was much reduced in granule cells cultured in CDM as compared to those maintained in the presence of serum. These data would indicate that in CDM the differentiation of granule neurons is not complete, while that of GABAergic neurons is not greatly affected. Whether the diminished differentiation of granule cells must be attributed only to serum deprivation or also to other differences in the composition of the culture medium remains to be established. /sup 3/H-GABA was avidly taken up also by a population of cells which were not recognized by antibodies raised against GFAP, glutamate decarboxylase, and microtubule-associated protein 2. These cells have been characterized as bipotential precursors of oligodendrocytes and of a subpopulation of astrocytes bearing a stellate shape and capable of high-affinity /sup 3/H-GABA uptake.

  20. Conditioned medium from mesenchymal stem cells induces cell death in organotypic cultures of rat hippocampus and aggravates lesion in a model of oxygen and glucose deprivation.

    Science.gov (United States)

    Horn, Ana Paula; Frozza, Rudimar Luiz; Grudzinski, Patrícia Benke; Gerhardt, Daniéli; Hoppe, Juliana Bender; Bruno, Alessandra Nejar; Chagastelles, Pedro; Nardi, Nance Beyer; Lenz, Guido; Salbego, Christianne

    2009-01-01

    Cell therapy using bone marrow-derived mesenchymal stem cells (MSC) seems to be a new alternative for the treatment of neurological diseases, including stroke. In order to investigate the response of hippocampal tissue to factors secreted by MSC and if these factors are neuroprotective in a model of oxygen and glucose deprivation (OGD), we used organotypic hippocampal cultures exposed to conditioned medium from bone marrow-derived MSC. Our results suggest that the conditioned medium obtained from these cells aggravates lesion caused by OGD. In addition, the presence of the conditioned medium alone was toxic mainly to cells in the CA1, CA2 and CA3 areas of the hippocampal organotypic culture even in basal conditions. GABA stimulation and NMDA and AMPA receptors antagonists were able to reduce propidium iodide staining, suggesting that the cell death induced by the toxic factors secreted by MSC could involve these receptors.

  1. Visualizing medium and biodistribution in complex cell culture bioreactors using in vivo imaging.

    Science.gov (United States)

    Ratcliffe, E; Thomas, R J; Stacey, A J

    2014-01-01

    There is a dearth of technology and methods to aid process characterization, control and scale-up of complex culture platforms that provide niche micro-environments for some stem cell-based products. We have demonstrated a novel use of 3d in vivo imaging systems to visualize medium flow and cell distribution within a complex culture platform (hollow fiber bioreactor) to aid characterization of potential spatial heterogeneity and identify potential routes of bioreactor failure or sources of variability. This can then aid process characterization and control of such systems with a view to scale-up. Two potential sources of variation were observed with multiple bioreactors repeatedly imaged using two different imaging systems: shortcutting of medium between adjacent inlet and outlet ports with the potential to create medium gradients within the bioreactor, and localization of bioluminescent murine 4T1-luc2 cells upon inoculation with the potential to create variable seeding densities at different points within the cell growth chamber. The ability of the imaging technique to identify these key operational bioreactor characteristics demonstrates an emerging technique in troubleshooting and engineering optimization of bioreactor performance. © 2013 American Institute of Chemical Engineers.

  2. Morphological differentiation between S. mutans and S. sobrinus on modified SB-20 culture medium.

    Science.gov (United States)

    Saravia, Marta Estela; Nelson-Filho, Paulo; Ito, Izabel Yoko; da Silva, Léa Assed Bezerra; da Silva, Raquel Assed Bezerra; Emilson, Claes-Göran

    2011-01-20

    Due to the major role of Streptococcus mutans and Streptococcus sobrinus in the etiology of dental caries, it is important to use culture media that allow for differentiating these bacterial species. The aim of this study was to evaluate the suitability of a modified SB-20 culture medium (SB-20M) for the isolation and morphological differentiation of S. mutans and S. sobrinus, compared to biochemical identification (biotyping). Saliva samples were collected using the spatula method from 145 children, seeded on plates containing the SB-20M, in which sucrose was replaced by coarse granular cane sugar, and incubated in microaerophilia at 37°C during 72 h. Identification of the microorganisms was performed under stereomicroscopy based on colony morphology of 4904 colonies. The morphological identification was examined by biochemical tests of 94 randomly selected colonies with the macroscopic characteristic of S. mutans and S. sobrinus using sugar fermentation, resistance to bacitracin and production of hydrogen peroxide. There was no statistically significant difference (p>0.05) between morphological identification in the SB-20M medium and biochemical identification (biotyping). Biotyping confirmed that S. mutans and S. sobrinus colonies were correctly characterized in the SB-20M in 95.8% and 95.5% of the cases, respectively. Of the mutans streptococci detected in the children 98% were S. mutans and 2% S. sobrinus. The SB-20M medium is reliable for detection and direct morphological identification of S. mutans and S. sobrinus.

  3. Culture filtrate antigens and allergens of Epicoccum nigrum cultivated in modified semi-synthetic medium.

    Science.gov (United States)

    Bisht, Vandana; Singh, Bhanu Pratap; Kumar, Raj; Arora, Naveen; Sridhara, Susheela

    2002-05-01

    Epicoccum nigrum (EN) is an important fungal allergen for nasobronchial allergy. Fungal extracts should contain all the relevant allergen components from spores, mycelium and culture medium for the purpose of allergy diagnosis and therapy. EN extract from spore-mycelial mass has been standardized, but the culture filtrate (CF) allergens of EN have not been studied as EN grows poorly in synthetic medium. The objective of the present study was to obtain a standard CF extract of EN by cultivating the source material in a modified semi-synthetic medium and to compare this with the EN cellular extract. Sabouraud's medium containing yeast extract (50 mg/l) was filtered using 10-kDa cut-off membrane and the lower molecular mass media components were used to cultivate EN. The CF obtained after removing the spore-mycelia was dialyzed to remove media components. The CF extract was characterized by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot. It was compared with EN spore-mycelial extract by enzyme-linked immunosorbent assay (ELISA), ELISA inhibition and by intradermal testing on allergy patients. The CF extract of EN resolved into 30 protein bands on SDS-PAGE. About 27 IgG bands were detected using anti-EN rabbit antibodies and 12 IgE bands by EN-sensitive pooled patients' sera. Periodate modification of CF proteins showed that the carbohydrate moieties are not important for IgE binding. Protein components of 26, 34 and 43 kDa were recognized as the major CF allergens. Three different batches of CF extract required 7.5-9 ng of self protein for 50% inhibition of binding to anti-EN rabbit antibodies in ELISA. Intradermal testing with CF extract showed comparable allergenic potency to standardized EN spore-mycelial extract, although it contained some allergenic proteins in higher amounts as compared to the spore-mycelial extract. In summary, the semi-synthetic medium has been suitably modified for obtaining EN CF antigens. This medium can

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

    Science.gov (United States)

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

    2016-10-01

    Does embryo culture medium influence pregnancy and perinatal outcome in IVF? Embryo culture media used in IVF affect treatment efficacy and the birthweight of newborns. A wide variety of culture media for human preimplantation embryos in IVF/ICSI treatments currently exists. It is unknown which medium is best in terms of clinical outcomes. Furthermore, it has been suggested that the culture medium used for the in vitro culture of embryos affects birthweight, but this has never been demonstrated by large randomized trials. We conducted a multicenter, double-blind RCT comparing the use of HTF and G5 embryo culture media in IVF. Between July 2010 and May 2012, 836 couples (419 in the HTF group and 417 in the G5 group) were included. The allocated medium (1:1 allocation) was used in all treatment cycles a couple received within 1 year after randomization, including possible transfers with frozen-thawed embryos. The primary outcome was live birth rate. Couples that were scheduled for an IVF or an ICSI treatment at one of the six participating centers in the Netherlands or their affiliated clinics. The live birth rate was higher, albeit nonsignificantly, in couples assigned to G5 than in couples assigned to HTF (44.1% (184/417) versus 37.9% (159/419); RR: 1.2; 95% confidence interval (CI): 0.99-1.37; P = 0.08). Number of utilizable embryos per cycle (2.8 ± 2.3 versus 2.3 ± 1.8; P HTF. Of the 383 live born children in this trial, birthweight data from 380 children (300 singletons (G5: 163, HTF: 137) and 80 twin children (G5: 38, HTF: 42)) were retrieved. Birthweight was significantly lower in the G5 group compared with the HTF group, with a mean difference of 158 g (95% CI: 42-275 g; P = 0.008). More singletons were born preterm in the G5 group (8.6% (14/163) versus 2.2% (3/137), but singleton birthweight adjusted for gestational age and gender (z-score) was also lower in the G5 than in the HTF group (-0.13 ± 0.08 versus 0.17 ± 0.08; P = 0.008). This study was powered

  5. Acute toxicity bioassays using Daphnia magna Straus (Cladocera, Daphniidae maintained in a modified culture medium

    Directory of Open Access Journals (Sweden)

    Mónica Núñez

    2013-05-01

    Full Text Available Daphnia magna is a test organism used in ecotoxicological assays of freshwater; however, traditional culture systems for this organism could result expensive, for that the aim of this research was to developed a new economic culture medium. With this purpose, 10 strains of D. magna were isolated, their population development was evaluated by total count of organisms and pregnant females using 3 different culture media: (A alfalfa juice, (B solved yeast and (C a mixture of alfalfa juice plus solved yeast. Successful development of 4 strains was observed in the A medium, but the same strains failed to survive in the B and the C media. The 24h and 48h EC50 average values in acute ecotoxicological assays with potassium dicromate were 0,4045 mg/L ± 0,0389 and 0,1857 mg/L ± 0,0072 respectively. Also, acute ecotoxicological assays with these 4 strains were performed using potassium cyanide, which is a toxic reactive frequently used in mining operations. In this case 24h EC50 value was 1,5388 mg/L ± 0,1146 and 48h EC50 values were 0,6359 mg/L ± 0,0516. 48h EC50 values were lower than the cyanide permissible effluent values established by the Energy and Mining Authority.

  6. Crustacean peptidergic neurons in culture show immediate outgrowth in simple medium.

    Science.gov (United States)

    Cooke, I; Graf, R; Grau, S; Haylett, B; Meyers, D; Ruben, P

    1989-01-01

    The survival and outgrowth of neurons in culture has usually required conditioning factors. We now report that crustacean neurons, taken from the peptidergic neurosecretory system of the eyestalk of crabs (Cardisoma carnifex) and lobsters (Panulirus marginatus), show immediate outgrowth, sustained for a week or more, in defined medium as simple as physiological saline with glucose and glutamine. The neurons show peptide hormone immunoreactivity that is prominent at growth cones, exhibit differences in form correlated with their immunoreactivity, release peptides to the medium, and have voltage-dependent currents, including a well-sustained Ca current. Cd blocks secretion, growth, and the Ca current. Peptidergic secretory neurons may be able to utilize existing membrane from their store of granules and already active synthetic, transport, and secretory mechanisms for immediate outgrowth.

  7. Wal-Mart’s Corporate Culture and the Suggestions for China’s Retailing Industry in Culture-Building

    Institute of Scientific and Technical Information of China (English)

    孙德洋

    2013-01-01

    With sales at a quarter of a trillion and over 1.3 million employees, Wal-Mart has been the biggest retailing success ever before. A lot of Wal-Mart’s success was attributed to the strong and pervasive culture at the company.China’s retailing industry should really learn something about corporate culture-building from this excellent enterprise, so that one day it would become a strong competitor both home and abroad in retailing industry.

  8. Development of an alternative culture medium for the selective enumeration of Lactobacillus casei in fermented milk.

    Science.gov (United States)

    Colombo, Monique; de Oliveira, Aline Evelyn Zimmermann; de Carvalho, Antonio Fernandes; Nero, Luís Augusto

    2014-05-01

    Monitoring the populations of probiotic strains of the species Lactobacillus casei in food is required by food industries in order to assure that a minimum concentration of these organisms will be ingested by consumers. In this context, Petrifilm™ AC plates can be used along with selective culture media to allow the enumeration of specific groups of lactic acid bacteria. The present study aimed to assess chemical substances as selective agents for Lb. casei in order to propose a selective culture medium to be used with Petrifilm™ AC plates as an alternative protocol for the enumeration of probiotic strains of this species in fermented milk. Twenty-six probiotic and starter cultures (including six strains of Lb. casei) were plated on de Man Rogosa and Sharpe (MRS) agar with distinct concentrations of nalidixic acid, bile, lithium chloride, metronidazole, sodium propionate, and vancomycin. Vancomycin at 10 mg/L demonstrated selective activity for Lb. casei. In addition, 2,3,5-triphenyltetrazolium chlorine was identified as a compound that did not inhibit Lb. casei, and Petrifilm™ AC plates used with MRS and vancomycin at 10 mg/L (MRS-V) demonstrated more colonies of this organism when incubated under anaerobic conditions than aerobic conditions. Acidophilus milk and yoghurt were prepared, added to Lb. casei strains, and stored at 4 °C. Lb. casei populations were monitored using MRS-V and MRTLV by conventional plating and associated with Petrifilm™ AC plates. All correlation indices between counts obtained by conventional plating and Petrifilm™ AC were significant (p enumeration of Lb. casei strains in fermented milk. However, the selective potential of this culture medium must be evaluated considering the specific strains of Lb. casei and the starter cultures inoculated in the fermented milk that requires monitoring.

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

    Directory of Open Access Journals (Sweden)

    Virginie eBonnamain

    2013-12-01

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

  10. DETECTION AND CHARACTERISTIC OF FLAT ERYTHROID COLONIES IN SEMISOLID CULTURAL MEDIUMS

    Directory of Open Access Journals (Sweden)

    M. D. Kuchma

    2014-06-01

    Full Text Available It have been shown that progenitor cells of cord blood, bone marrow and «mobilized» peripheral blood in semisolid mediums gave flat erythroid colonies. These colonies are able to form one or more red centers on the 14th day of cultivation and get a big size that evidence about high proliferative activity and resemble granulocyte, erythrocyte, monocyte/macrophage, megakaryocyte colony-forming units. However 92% of the cells of flat colonies express CD235. It shows that the colonies are erythroid, although colony morphology differs from burstoforming erythroid units and erythroid colony forming units. Their occurrence probability in methylcellulose-containing medium is 2,5%±1%, that is significantly lower than in agar- containing medium (58%±4,8%. Thus, we suggested that flat colonies should be counted separately or they should be ascribed as BFU-E.

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

    Science.gov (United States)

    Bliesner, Anna; Liedtke, Christa; Rohn, Holger

    2014-05-15

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

  12. Semi-selective culture medium for Exserohilum turcicum isolation from corn seeds

    Directory of Open Access Journals (Sweden)

    Roberto Luis De Rossi

    2014-06-01

    Full Text Available Northern corn leaf blight, caused by Exserohilum turcicum (Et, is a disease of widespread occurrence in regions where corn, sweetcorn and popcorn are grown. This disease has great potential to cause damage and has been studied for years, but the association of its causal agent with seeds remains unconfirmed. Thus, the availability of a sensitive method to detect and quantify the inoculum in seeds, even at low incidence, is essential. The aim of this study was to develop a method to detect and quantify the presence of the fungus infecting and infesting corn and popcorn seeds. Artificially and naturally infected seeds were employed to develop the medium. The semi-selective medium was composed of carbendazim (active ingredient (60 mg/L, captan (30 mg/L, streptomycin sulfate (500 mg/L and neomycin sulfate (600 mg/L aggregated to the medium lactose casein hydrolysate agar medium. By using this, Et was detected in naturally infected corn seeds, showing 0.124% incidence, in four out of ten analyzed samples. In addition, 1.04 conidia were detected per infested seed. By means of isolation, pathogenicity test, morphological characterization and comparison with descriptions of the species in the literature, the fungus isolated from the seeds was confirmed to be Et. Both infection and infestation were considered low; thus, for studies of Et detection in corn seeds, the use of semi-selective medium and more than 1,200 seeds/sample is suggested.

  13. [Geothermal water as part of culture medium and morpho-physiological properties of Saccharomyces cerevisiae].

    Science.gov (United States)

    Abramov, Sh A; Kotenko, S Ts; Khalilova, E A; Kisrieva, Iu S

    1999-01-01

    Morphophysiological changes in Saccharomyces cerevisiae gamma-503 cells cultivated in nutrient media containing geothermal water as a source of mineral substances were studied. The optimal mineralization of the medium was found to be 4.0 g/l, supplemented with 2.6 g/l (NH4)2HPO4. These conditions provided active growth and development of the culture with high yields of the biomass and the maximal enzymatic activity. Differences in cellular structures at certain stages of metabolism were demonstrated.

  14. Legionella pneumophila Arthritis: use of medium specific for Mycobacteria for isolation of L. pneumophila in culture of articular fluid specimens.

    Science.gov (United States)

    Bemer, Pascale; Leautez, Sophie; Ninin, Emmanuelle; Jarraud, Sophie; Raffi, François; Drugeon, Henri

    2002-07-01

    We report the first case, to our knowledge, of acute purulent arthritis due to Legionella pneumophila in an immunosuppressed patient. L. pneumophila was isolated from samples of blood and articular fluid cultured with use of medium specific for mycobacteria (Bactec 13A medium).

  15. Improving the culturability of freshwater bacteria using FW70, a low-nutrient solid medium amended with sodium pyruvate.

    Science.gov (United States)

    Imazaki, Iori; Kobori, Youichi

    2010-04-01

    Bacterial culture based on the use of plate media is an effective method for investigating bacterial populations in the environment. To improve the culturability of bacteria from freshwater lakes, we developed a new medium, FW70, which contains sodium pyruvate, casamino acids, and lake water and is solidified using gellan gum. To test the importance of these components, we prepared a series of media in which one or more of the components was absent. Water was sampled 31 times from 3 Japanese lakes and was passed through a membrane filter (pore size = 0.45 microm) to remove fast-growing microbes before the water was spread onto the plates. In most cases, significantly larger numbers of bacterial colonies were detected on FW70 than on other media. Furthermore, to test the practicality of FW70, we compared it with standard nutrient agar and R2A agar. In all cases, the culturability was significantly greater on FW70 than on standard nutrient agar or R2A agar. Some isolates recovered by means of FW70 belonged to bacteria that had not previously been classified. These results suggest that FW70 improves the culturability of freshwater bacteria and can be used for the isolation of novel bacteria as a result of the filtration step.

  16. Influence of culture medium growth variables on Ganoderma lucidum exopolysaccharides structural features.

    Science.gov (United States)

    Fraga, Irene; Coutinho, João; Bezerra, Rui M; Dias, Albino A; Marques, Guilhermina; Nunes, Fernando M

    2014-10-13

    In this work the effect of carbon and nitrogen levels and initial pH of the wheat extract culture medium of submerged culture of Ganoderma lucidum on the amount, purity and structural features of exopolysaccharides (EPS) were studied. A low peptone level (1.65 g L(-1)) favored mycelium biomass, EPS purity, but a higher supply of peptone (4.80 g L(-1)) is needed for maximum EPS production. The carbohydrate composition of the EPS and structural features also changed significantly according to the different growing conditions, being observed significant differences in the (1 → 3)/(1 → 4)-Glcp ratio and also on the branching degree of EPS. As the biological activities of EPS are highly dependent on the polysaccharide structural features, this variability can have implications on the EPS biological activities, but can also be used advantageously to produce tailor made polysaccharides with specific applications.

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

    Directory of Open Access Journals (Sweden)

    Enrique Salas Barbosa

    2005-04-01

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

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

    Directory of Open Access Journals (Sweden)

    YUSNITA

    2005-06-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2015-05-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1998-11-01

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

  1. Hyperconcentrated Sweet Whey, a New Culture Medium That Enhances Propionibacterium freudenreichii Stress Tolerance

    Science.gov (United States)

    Huang, Song; Rabah, Houem; Jardin, Julien; Briard-Bion, Valérie; Parayre, Sandrine; Maillard, Marie-Bernadette; Le Loir, Yves; Schuck, Pierre; Jeantet, Romain

    2016-01-01

    ABSTRACT Propionibacterium freudenreichii is used as a cheese-ripening starter and as a probiotic. Its reported physiological effects at the gut level, including modulation of bifidobacteria, colon epithelial cell proliferation and apoptosis, and intestinal inflammation, rely on active metabolism in situ. Survival and activity are thus key factors determining its efficacy, creating stress adaptation and tolerance bottlenecks for probiotic applications. Growth media and growth conditions determine tolerance acquisition. We investigated the possibility of using sweet whey, a dairy by-product, to sustain P. freudenreichii growth. It was used at different concentrations (dry matter) as a culture medium. Using hyperconcentrated sweet whey led to enhanced multistress tolerance acquisition, overexpression of key stress proteins, and accumulation of intracellular storage molecules and compatible solutes, as well as enhanced survival upon spray drying. A simplified process from growth to spray drying of propionibacteria was developed using sweet whey as a 2-in-1 medium to both culture P. freudenreichii and protect it from heat and osmotic injury without harvesting and washing steps. As spray drying is far cheaper and more energy efficient than freeze-drying, this work opens new perspectives for the sustainable development of new starter and probiotic preparations with enhanced robustness. IMPORTANCE In this study, we demonstrate that sweet whey, a dairy industry by-product, not only allows the growth of probiotic dairy propionibacteria, but also triggers a multitolerance response through osmoadaptation and general stress response. We also show that propionibacteria accumulate compatible solutes under these culture conditions, which might account for the limited loss of viability after spray drying. This work opens new perspectives for more energy-efficient production of dairy starters and probiotics. PMID:27235433

  2. Performance evaluation of CHO-K1 cell in culture medium supplemented with hemolymph

    Directory of Open Access Journals (Sweden)

    Tássia Raffoul

    2005-06-01

    Full Text Available The aim of this work was to evaluate the potential of hemolymph utilization as a culture medium supplement to cultivate the animal cell CHO-K1. For this purpose 1% v/v of hemolymph was added to DMEM medium containing 10% v/v of FBS and 1 or 4.5 g/L of glucose. The culture was grown in spinner flasks incubated in a 10% v/v CO2 environment, at 37ºC, with the Cytodex 1 microcarrier. Comparing the results obtained from the culture with hemolymph against those without hemolymph, a positive influence of the hemolymph was observed, as the experiment with hemolymph presented a 52% higher cell concentration and a higher productivity of up to 40%.Desenvolvimento de meios de cultura isentos de soro fetal bovino (SFB é uma das grandes prioridades de pesquisa em desenvolvimento de processos com célula animal. O objetivo do presente trabalho foi realizar uma análise do potencial de uso da hemolinfa como suplemento do meio utilizado no cultivo da célula animal ancorante CHO-K1. Para isso, foi adicionado 1% v/v de extrato de hemolinfa ao meio DMEM contendo 10% v/v de SFB e 1,0 ou 4,5 g/L de glicose. O cultivo foi realizado em frascos tipo spinner em um ambiente de 10% v/v de CO2, a 37ºC, utilizando o microcarregador Cytodex 1. Comparando os resultados obtidos no ensaio com hemolinfa com um sem hemolinfa pode-se notar uma influência positiva da hemolinfa no cultivo, já que o ensaio com hemolinfa apresentou uma concentração máxima de células 52% maior e uma produtividade máxima de até 40% maior.

  3. The effect of chemically defined medium on spontaneous calcium signaling of in situ chondrocytes during long-term culture.

    Science.gov (United States)

    Zhou, Yilu; Park, Miri; Cheung, Enoch; Wang, Liyun; Lu, X Lucas

    2015-04-13

    Chemically defined serum-free medium has been shown to better maintain the mechanical integrity of articular cartilage explants than serum-supplemented medium during long-term in vitro culture, but little is known about its effect on cellular mechanisms. We hypothesized that the chemically defined culture medium could regulate the spontaneous calcium signaling of in situ chondrocytes, which may modulate the cellular metabolic activities. Bovine cartilage explants were cultured in chemically defined serum-free or serum-supplemented medium for four weeks. The spontaneous intracellular calcium ([Ca(2+)]i) signaling of in situ chondrocytes was longitudinally measured together along with the biomechanical properties of the explants. The spontaneous [Ca(2+)]i oscillations in chondrocytes were enhanced at the initial exposure of serum-supplemented medium, but were significantly dampened afterwards. In contrast, cartilage explants in chemically defined medium preserved the level of calcium signaling, and showed more responsive cells with higher and more frequent [Ca(2+)]i peaks throughout the four week culture in comparison to those in serum medium. Regardless of the culture medium that the explants were exposed, a positive correlation was detected between the [Ca(2+)]i responsive rate and the stiffness of cartilage (Spearman's rank correlation coefficient=0.762). A stable pattern of [Ca(2+)]i peaks was revealed for each chondrocyte, i.e., the spatiotemporal features of [Ca(2+)]i peaks from a cell were highly consistent during the observation period (15 min). This study showed that the beneficial effect of chemically defined culture of cartilage explants is associated with the spontaneous [Ca(2+)]i signaling of chondrocytes in cartilage.

  4. Single Cell Protein Production by Saccharomyces cerevisiae Using an Optimized Culture Medium Composition in a Batch Submerged Bioprocess.

    Science.gov (United States)

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

    2016-08-01

    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.

  5. Production of Fusaric Acid by Fusarium spp. in Pure Culture and in Solid Medium Co-Cultures

    Directory of Open Access Journals (Sweden)

    Nadine Bohni

    2016-03-01

    Full Text Available The ability of fungi isolated from nails of patients suffering from onychomycosis to induce de novo production of bioactive compounds in co-culture was examined. Comparison between the metabolite profiles produced by Sarocladium strictum, by Fusarium oxysporum, and by these two species in co-culture revealed de novo induction of fusaric acid based on HRMS. Structure confirmation of this toxin, using sensitive microflow NMR, required only three 9-cm Petri dishes of fungal culture. A targeted metabolomics study based on UHPLC-HRMS confirmed that the production of fusaric acid was strain-dependent. Furthermore, the detected toxin levels suggested that onychomycosis-associated fungal strains of the F. oxysporum and F. fujikuroi species complexes are much more frequently producing fusaric acid, and in higher amount, than strains of the F. solani species complex. Fusarium strains producing no significant amounts of this compound in pure culture, were shown to de novo produce that compound when grown in co-culture. The role of fusaric acid in fungal virulence and defense is discussed.

  6. Production of Fusaric Acid by Fusarium spp. in Pure Culture and in Solid Medium Co-Cultures.

    Science.gov (United States)

    Bohni, Nadine; Hofstetter, Valérie; Gindro, Katia; Buyck, Bart; Schumpp, Olivier; Bertrand, Samuel; Monod, Michel; Wolfender, Jean-Luc

    2016-03-18

    The ability of fungi isolated from nails of patients suffering from onychomycosis to induce de novo production of bioactive compounds in co-culture was examined. Comparison between the metabolite profiles produced by Sarocladium strictum, by Fusarium oxysporum, and by these two species in co-culture revealed de novo induction of fusaric acid based on HRMS. Structure confirmation of this toxin, using sensitive microflow NMR, required only three 9-cm Petri dishes of fungal culture. A targeted metabolomics study based on UHPLC-HRMS confirmed that the production of fusaric acid was strain-dependent. Furthermore, the detected toxin levels suggested that onychomycosis-associated fungal strains of the F. oxysporum and F. fujikuroi species complexes are much more frequently producing fusaric acid, and in higher amount, than strains of the F. solani species complex. Fusarium strains producing no significant amounts of this compound in pure culture, were shown to de novo produce that compound when grown in co-culture. The role of fusaric acid in fungal virulence and defense is discussed.

  7. Production of Normal Mammalian Organ Culture Using a Medium Containing Mem-Alpha, Leibovitz L 15, Glucose Galactose Fructose

    Science.gov (United States)

    Goodwin, Thomas J. (Inventor); Wolf, David A. (Inventor); Spaulding, Glenn F. (Inventor); Prewett, Tacey L. (Inventor)

    1999-01-01

    Normal mammalian tissue and the culturing process has been developed for the three groups of organ, structural and blood tissue. The cells are grown in vitro under micro- gravity culture conditions and form three dimensional cells aggregates with normal cell function. The microgravity culture conditions may be microgravity or simulated microgravity created in a horizontal rotating wall culture vessel. The medium used for culturing the cells, especially a mixture of epithelial and mesenchymal cells contains a mixture of Mem-alpha and Leibovits L15 supplemented with glucose, galactose and fructose.

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

    Science.gov (United States)

    Cote, Gerald L.

    1993-01-01

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

  9. Evaluation of Lo"wenstein-Jensen Medium Culture,MGIT 960 Culture and Different Specimen Types inDiagnosis of Bone and Joint Tuberculosis

    Institute of Scientific and Technical Information of China (English)

    Guirong Wang[1; Weijie Dong[2; Liping Zhao[1; Xia Yu[1; Suting Chen[1; Yuhong Fu[1; Shibing Qin[1; Hairong Huang[1

    2015-01-01

    Objective: The aim of this study was to evaluate L-J (Lo"wenstein-Jensen) medium culture, MGIT 960 culture anddifferent specimen types in diagnosis of BJTB (bone and joint tuberculosis). Methods:: Specimens of pus, caseous necrosis,tuberculous granuloma and sequestrum were collected from 52 BJTB patients. All specimens were cultured using both MGIT 960system and L-J medium; and all pus were amplified using real-time PCR to detect the presence of M. tuberculosis DNA. KeyFindings: A total of 191 specimens were collected. Granuloma had better chance to produce positive outcomes by L-J mediumculture, while for sequestrum MGIT 960 culture had higher yield, but there was no significant difference in the recovery rates amongdifferent types of specimen either by L-J culture (Z2 = 0.638, P = 0.888) or by MGIT960 culture (Z2 = 1.399, P = 0.706). MGIT960culture had significantly higher recovery rate than L-J culture, With a combined culture and PCR-based test, the recovery rate of pusspecimen was significantly higher than that of either method alone (P 〈 0.05). Conclusion: MGIT 960 culture is superior to L-Jculture in BJTB diagnosis; pus, sequestrum, granuloma and caseous necrosis are usable specimen for mycobacterial culture;combination of culture and molecular techniques can provide a better diagnostic significance.

  10. Medium Optimization for Exopolysaccharide Production in Liquid Culture of Endophytic Fungus Berkleasmium sp. Dzf12

    Directory of Open Access Journals (Sweden)

    Peiqin Li

    2012-09-01

    Full Text Available Berkleasmium sp. Dzf12, an endophytic fungus from Dioscorea zingiberensis, is a high producer of spirobisnaphthalenes with various bioactivities. The exopolysaccharide (EPS produced by this fungus also shows excellent antioxidant activity. In this study, the experimental designs based on statistics were employed to evaluate and optimize the medium for EPS production in liquid culture of Berkleasmium sp. Dzf12. For increasing EPS yield, the concentrations of glucose, peptone, KH2PO4, MgSO4∙7H2O and FeSO4∙7H2O in medium were optimized using response surface methodology (RSM. Both the fractional factorial design (FFD and central composite design (CCD were applied to optimize the main factors which significantly affected EPS production. The concentrations of glucose, peptone and MgSO4∙7H2O were found to be the main effective factors for EPS production by FFD experimental analysis. Based on the further CCD optimization and RSM analysis, a quadratic polynomial regression equation was derived from the EPS yield and three variables. Statistical analysis showed the polynomial regression model was in good agreement with the experimental results with the determination coefficient (adj-R2 as 0.9434. By solving the quadratic regression equation, the optimal concentrations of glucose, peptone and MgSO4∙7H2O for EPS production were determined as 63.80, 20.76 and 2.74 g/L, respectively. Under the optimum conditions, the predicted EPS yield reached the maximum (13.22 g/L. Verification experiment confirmed the validity with the actual EPS yield as 13.97 g/L, which was 6.29-fold in comparison with that (2.22 g/L in the original basal medium. The results provide the support data for EPS production in large scale and also speed up the application of Berkleasmium sp. Dzf12.

  11. Characteristics of plasma in culture medium generated by positive pulse voltage and effects of organic compounds on its characteristics

    Science.gov (United States)

    Sato, Y.; Sato, T.; Yoshino, D.

    2016-12-01

    We describe a positive pulse voltage method for generating plasma in culture medium with a composition similar to biological fluids. We also describe the plasma’s characteristics, liquid quality, and the effect of organic compounds in the culture medium on the plasma characteristics through comparisons to a solution containing inorganic salts at the same concentrations as in the culture medium. Light emission with Na and OH spectra was observed within a vapor bubble produced by Joule heating at the tip of the electrode. A downward thermal flow and shock wave were caused by the behavior of the vapor bubble. The culture medium pH gradually increased from 7.9 to 8.3 over the discharge time of 300 s. H2O2 was generated 1.1 mg l-1 in the culture medium after discharge for 300 s, and this value was 0.5 mg l-1 lower than the inorganic salts solution which does not contain organic compounds. This study provides important data that will help facilitate more widespread application of plasma medicine.

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

    Energy Technology Data Exchange (ETDEWEB)

    Eberbeck, D; Zirpel, P; Trahms, L [Physikalisch-Technische Bundesanstalt, Abbestrasse 2-12, 10587 Berlin (Germany); Kettering, M; Hilger, I [Institute of Diagnostic and Interventional Radiology, University Hospital Jena, Erlanger Allee 101, 07747 Jena (Germany); Bergemann, C, E-mail: dietmar.eberbeck@ptb.d [Chemicell GmbH, Eresburgstrasse 22-23, 12103 Berlin (Germany)

    2010-10-13

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

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

    Science.gov (United States)

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

    2013-01-01

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

  14. Optimization of the basal medium for improving production and secretion of taxanes from suspension cell culture of Taxus baccata L

    Directory of Open Access Journals (Sweden)

    Kajani Abolghasem

    2012-10-01

    Full Text Available Abstract Background and purpose of the study Taxol is one of the most effective anticancer drugs that isolated from Taxus sp. due to the slow growth of Taxus trees and low concentration of Taxol in the tissues, the biotechnological approaches especially plant cell culture have been considered to produce Taxol in commercial scale. Methods We investigated the effects of basal medium type used in culture media on production of Taxol and other taxane compounds from cell suspension culture of T. baccata L. Briefly, five commonly basal media including Gamborg, Murashige and Skoog, Woody Plant, Schenk and Hildebrandt, and Driver and Kuniyuki medium were used for preparing separate suspension culture media. The intra- and extra-cellular yields of taxanes were analyzed by using HPLC after 21 days period of culturing. Results The yields of taxanes were significantly different for the cultures prepared by different basal media. Moreover, the effects of basal medium on the yield of products differed for varius taxane compounds. Maximum yields of Baccatin III (10.03 mgl-1 and 10-deacetyl baccatin III (4.2 mgl-1 were achieved from the DKW basal media, but the yield of Taxol was maximum (16.58 mgl-1 in the WPM basal media. Furthermore, the secretion of taxanes from the cells into medium was also considerably affected by the type of basal medium. The maximum extra-cellular yield of Taxol (7.81 mgl-1, Baccatin III (5.0 mgl-1, and 10-deacetyl baccatin III (1.45 mgl-1 were also obtained by using DKW basal medium that were significantly higher than those obtained from other culture media.

  15. Optimization of the basal medium for improving production and secretion of taxanes from suspension cell culture of Taxus baccata L

    Directory of Open Access Journals (Sweden)

    Abolghasem Abbasi Kajani

    2012-10-01

    Full Text Available Background and purpose of the study Taxol is one of the most effective anticancer drugs that isolated from Taxus sp. due to the slow growth of Taxus trees and low concentration of Taxol in the tissues, the biotechnological approaches especially plant cell culture have been considered to produce Taxol in commercial scale.MethodsWe investigated the effects of basal medium type used in culture media on production of Taxol and other taxane compounds from cell suspension culture of T. baccata L. Briefly, five commonly basal media including Gamborg, Murashige and Skoog, Woody Plant, Schenk and Hildebrandt, and Driver and Kuniyuki medium were used for preparing separate suspension culture media. The intra- and extra-cellular yields of taxanes were analyzed by using HPLC after 21 days period of culturing.ResultsThe yields of taxanes were significantly different for the cultures prepared by different basal media. Moreover, the effects of basal medium on the yield of products differed for varius taxane compounds. Maximum yields of Baccatin III (10.03 mgl-1 and 10-deacetyl baccatin III (4.2 mgl-1 were achieved from the DKW basal media, but the yield of Taxol was maximum (16.58 mgl-1 in the WPM basal media. Furthermore, the secretion of taxanes from the cells into medium was also considerably affected by the type of basal medium. The maximum extra-cellular yield of Taxol (7.81 mgl-1, Baccatin III (5.0 mgl-1, and 10-deacetyl baccatin III (1.45 mgl-1 were also obtained by using DKW basal medium that were significantly higher than those obtained from other culture media.

  16. Optimization of the Liquid Culture Medium Composition to Obtain the Mycelium of Agaricus bisporus Rich in Essential Minerals.

    Science.gov (United States)

    Krakowska, Agata; Reczyński, Witold; Muszyńska, Bożena

    2016-09-01

    Agaricus bisporus species (J.E. Lange) Imbach one of the most popular Basidiomycota species was chosen for the research because of its dietary and medicinal value. The presented herein studies included determination of essential mineral accumulation level in the mycelium of A. bisporus, cultivated on liquid cultures in the medium supplemented with addition of the chosen metals' salts. Quantitative analyses of Zn, Cu, Mg, and Fe in liquid cultures made it possible to determine the relationship between accumulation of the selected mineral in A. bisporus mycelium and the culture conditions. Monitoring of the liquid cultures and determination of the elements' concentrations in mycelium of A. bisporus were performed using the flame technique of AAS method. Concentration of Zn in the mycelium, maintained in the medium with the addition of its salt, was in a very wide range from 95.9 to 4462.0 mg/g DW. In the analyzed A. bisporus mycelium, cultured in the medium enriched with copper salt, this metal concentration changed from 89.79 to 7491.50 mg/g DW; considering Mg in liquid cultured mycelium (medium with Mg addition), its concentration has changed from 0.32 to 10.55 mg/g DW. The medium enriched with iron salts has led to bioaccumulation of Fe in mycelia of A. bisporus. Determined Fe concentration was in the range from 0.62 to 161.28 mg/g DW. The proposed method of liquid A. bisporus culturing on medium enriched with the selected macro- and microelements in proper concentrations ratio have led to obtaining maximal growth of biomass, characterized by high efficiency of the mineral accumulation. As a result, a dietary component of increased nutritive value was obtained.

  17. Evidence for the involvement of nematocidal toxins of Purpureocillium lilacinum 6029 cultured on Karanja deoiled cake liquid medium.

    Science.gov (United States)

    Sharma, Abhishek; Sharma, Satyawati; Mittal, Aditya; Naik, S N

    2016-05-01

    In present study, in vitro nematocidal bioassays, FT-IR and HPLC analysis were employed to demonstrate the involvement of toxins of Purpureocillium lilacinum in killing root-knot nematodes (Meloidogyne incognita). During growth study, maximum mycelial biomass (10.52 g/l) in de-oiled Karanja cake medium was achieved on 8th day while complete mortality of nematodes was obtained by 6th day filtrate (FKSM). Maximum production of crude nematocidal toxin was recorded on 7th day suggesting that the toxin production was paralleled with growth of the fungus. The median lethal concentration (LC50) determined for the crude toxin from 6th day to 10th day ranged from 89.41 to 43.21 ppm. The median lethal time (LT50) for the crude toxin of FKSM was found to be 1.46 h. This is the first report of implementing a comparative infra-red spectroscopy coupled with HPLC analysis to predict the presence of nematocidal toxin in the fungal filtrate cultured on Karanja deoiled cake liquid medium.

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

    Directory of Open Access Journals (Sweden)

    S Lopez-Cazaux

    2006-02-01

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

  19. Comparison of dry medium culture plates for mesophilic aerobic bacteria in milk, ice cream, ham, and codfish fillet products.

    Science.gov (United States)

    Park, Junghyun; Kim, Myunghee

    2013-12-01

    This study was performed to compare the performance of Sanita-Kun dry medium culture plate with those of traditional culture medium and Petrifilm dry medium culture plate for the enumeration of the mesophilic aerobic bacteria in milk, ice cream, ham, and codfish fillet. Mesophilic aerobic bacteria were comparatively evaluated in milk, ice cream, ham, and codfish fillet using Sanita-Kun aerobic count (SAC), Petrifilm aerobic count (PAC), and traditional plate count agar (PCA) media. According to the results, all methods showed high correlations of 0.989~1.000 and no significant differences were observed for enumerating the mesophilic aerobic bacteria in the tested food products. SAC method was easier to perform and count colonies efficiently as compared to the PCA and PAC methods. Therefore, we concluded that the SAC method offers an acceptable alternative to the PCA and PAC methods for counting the mesophilic aerobic bacteria in milk, ice cream, ham, and codfish fillet products.

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

    Directory of Open Access Journals (Sweden)

    Grozeva Stanislava

    2014-01-01

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

  1. Quantitative determination of VEGF165 in cell culture medium by aptamer sandwich based chemiluminescence assay.

    Science.gov (United States)

    Shan, Siwen; He, Ziyi; Mao, Sifeng; Jie, Mingsha; Yi, Linglu; Lin, Jin-Ming

    2017-08-15

    In this work, we have developed a sensitive and selective chemiluminescence (CL) assay for vascular endothelial growth factor (VEGF165) quantitative detection based on two specific VEGF165 binding aptamers (Apt). VEGF is a predominant biomarker in cancer angiogenesis, and sensitive detection method of VEGF are highly demanded for both academic study and clinical diagnosis of multiple cancers. In our experiment, VEGF165 was captured in a sandwich structure assembled by two binding aptamers, one capture aptamer was immobilized on streptavidin-coated magnetic beads (MBs) and another VEGF-binding aptamer was labeled by biotin for further phosphatase conjunction. After Apt-VEGF-Apt sandwich was formed on MBs surface, alkaline phosphatase (ALP) was modified to the second aptamer to catalyze CL reaction. By applying 4-methoxy-4-(3-phosphatephenyl)-spiro-(1,2-dioxetane-3,2-adamantane) (AMPPD) as CL substrate, strong signal intensity was achieved. VEGF165 content as low as 1ng/mL was detected in standard spiked samples by our assay, and linear range of working curve was confirmed from 1 to 20ng/mL. Then our method was successfully applied for cell culture medium analysis and on-chip hypoxic HepG2-HUVEC co-culture model study with excellent accuracy equal to ELISA Kit. Our developed assay demonstrated an outstanding performance in VEGF165 quantification and may be further extended to clinical testing of important biomarkers as well as probing microchip cell culture model. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Synthesis of calcium oxalate crystals in culture medium irradiated with non-equilibrium atmospheric-pressure plasma

    Science.gov (United States)

    Kurake, Naoyuki; Tanaka, Hiromasa; Ishikawa, Kenji; Nakamura, Kae; Kajiyama, Hiroaki; Kikkawa, Fumitaka; Mizuno, Masaaki; Yamanishi, Yoko; Hori, Masaru

    2016-09-01

    Octahedral particulates several tens of microns in size were synthesized in a culture medium irradiated through contact with a plume of non-equilibrium atmospheric-pressure plasma (NEAPP). The particulates were identified in the crystalline phase as calcium oxalate dihydrate (COD). The original medium contained constituents such as NaCl, d-glucose, CaCl2, and NaHCO3 but not oxalate or oxalic acid. The oxalate was clearly synthesized and crystallized in the medium as thermodynamically unstable COD crystals after the NEAPP irradiation.

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

    Science.gov (United States)

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

    2016-05-01

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

  4. The effect of culture medium and carrier on explant culture of human limbal epithelium: A comparison of ultrastructure, keratin profile and gene expression.

    Science.gov (United States)

    Pathak, Meeta; Olstad, O K; Drolsum, Liv; Moe, Morten C; Smorodinova, Natalia; Kalasova, Sarka; Jirsova, Katerina; Nicolaissen, Bjørn; Noer, Agate

    2016-12-01

    Patients with limbal stem cell deficiency (LSCD) often experience pain and photophobia due to recurrent epithelial defects and chronic inflammation of the cornea. Successfully restoring a healthy corneal surface in these patients by transplantation of ex vivo expanded human limbal epithelial cells (LECs) may alleviate these symptoms and significantly improve their quality of life. The clinical outcome of transplantation is known to be influenced by the quality of transplanted cells. Presently, several different protocols for cultivation and transplantation of LECs are in use. However, no consensus on an optimal protocol exists. The aim of this study was to examine the effect of culture medium and carrier on the morphology, staining of selected keratins and global gene expression in ex vivo cultured LECs. Limbal biopsies from cadaveric donors were cultured for three weeks on human amniotic membrane (HAM) or on tissue culture coated plastic (PL) in either a complex medium (COM), containing recombinant growth factors, hormones, cholera toxin and fetal bovine serum, or in medium supplemented only with human serum (HS). The expanded LECs were examined by light microscopy (LM), transmission electron microscopy (TEM), immunohistochemistry (IHC) for keratins K3, K7, K8, K12, K13, K14, K15 and K19, as well as microarray and qRT-PCR analysis. The cultured LECs exhibited similar morphology and keratin staining on LM, TEM and IHC examination, regardless of the culture condition. The epithelium was multilayered, with cuboidal basal cells and flattened superficial cells. Cells were attached to each other by desmosomes. Adhesion complexes were observed between basal cells and the underlying carrier in LECs cultured on HAM, but not in LECs cultured on PL. GeneChip Human Gene 2.0 ST microarray (Affymetrix) analysis revealed that 18,653 transcripts were ≥2 fold up or downregulated (p ≤ 0.05). Cells cultured in the same medium (COM or HS) showed more similarities in gene

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

    Science.gov (United States)

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

    2010-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Kashyap R

    2010-01-01

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

  7. Induced biosynthesis of resveratrol and the prenylated stilbenoids arachidin-1 and arachidin-3 in hairy root cultures of peanut: Effects of culture medium and growth stage.

    Science.gov (United States)

    Condori, Jose; Sivakumar, Ganapathy; Hubstenberger, John; Dolan, Maureen C; Sobolev, Victor S; Medina-Bolivar, Fabricio

    2010-05-01

    Previously, we have shown that hairy root cultures of peanut provide a controlled, sustainable and scalable production system that can be induced to produce stilbenoids. However to leverage peanut hairy roots to study the biosynthesis of this polyphenolic biosynthetic pathway, growing conditions and elicitation kinetics of these tissue cultures must be defined and understood. To this end, a new peanut cv. Hull hairy root (line 3) that produces resveratrol and its prenylated analogues arachidin-1 and arachidin-3 upon sodium acetate-mediated elicitation was established. Two culture media were compared for impact on root growth and stilbenoid biosynthesis/secretion. The levels of ammonium, nitrate, phosphate and residual sugars were monitored along growth and elicitation period. A modified MS (MSV) medium resulted in higher root biomass when compared to B5 medium. The stilbenoid profile after elicitation varied depending on the age of the culture (6, 9, 12, and 15-day old). After elicitation at day 9 (exponential growth in MSV medium), over 90% of the total resveratrol, arachidin-1 and arachidin-3 accumulated in the medium. Our studies demonstrate the benefits of the hairy root culture system to study the biosynthesis of stilbenoids including valuable prenylated polyphenolic compounds.

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

    OpenAIRE

    Manju Pathak; Danik Martirosyan

    2012-01-01

    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 pre...

  9. Evaluation on Sensitivity of the Human Sperm Motility Assay for Detecting Endotoxin in Culture Medium

    Institute of Scientific and Technical Information of China (English)

    Wei-jie ZHU; Jing LI; Wen-hong ZHANG; Kang-shou YAO

    2003-01-01

    Objective To investigate the sensitivity of the human sperm motility assay for detecting endotoxin in culture medium Materials & Methods Motile sperm were separated and exposed to different concentrations of endotoxin (0.5 ng/mL, 1 ng/mL, 10 ng/mL, 1 000 ng/mL, 10 000 ng/mL, and 50 000 ng/mL), and sperm motility was determined after incubation. Effects of endotoxin on sperm motility in media without albumin were also examined. In addition, at the same concentrations of endotoxin (0.5 ng/mL, 1 ng/mL, and 10 ng/mL), the sensitivity of the human sperm motility assay was compared to those of 1-cell and 2-cell mouse embryo bioassays.Results At levels of 0.5 ng/mL~1 000 ng/mL endotoxin in media with 2 mg/mL albumin, sperm did not show significant change in motility during 24 h of incubation when compared with the control (P>0.05). However, the sperm motility was significantly inhibited at endotoxin dosages of 10 000 and 50 000 ng/mL. In the absence of albumin supplementation, at endotoxin levels of 50 000 ng/mL, and 1 000 ng/mL, there was a marked decrease in sperm motility compared with the control after 2 h or 8 h of incubation, respectively (P<0.01). In media containing 0.5 ng/mL and 1 ng/mL endotoxin, 1-cell and 2-cell mouse embryos had significantly reduced developmental rates in all developmental stages, and at the level of 10 ng/mL, the development of the embryos was arrested.Conclusion The human sperm motility assay could detect high levels of endotoxin in culture medium but its sensitivity to endotoxin would be inferior to that of the 1-cell or 2-cell mouse embryo bioassay. In the absence of albumin supplementation, the sensitivity of the sperm motility assay could be improved.

  10. Dopamine facilitates dendritic spine formation by cultured striatal medium spiny neurons through both D1 and D2 dopamine receptors.

    Science.gov (United States)

    Fasano, Caroline; Bourque, Marie-Josée; Lapointe, Gabriel; Leo, Damiana; Thibault, Dominic; Haber, Michael; Kortleven, Christian; Desgroseillers, Luc; Murai, Keith K; Trudeau, Louis-Éric

    2013-04-01

    Variations of dopamine (DA) levels induced by drugs of abuse or in the context of Parkinson's disease modulate the number of dendritic spines in medium spiny neurons (MSNs) of the striatum, showing that DA plays a major role in the structural plasticity of MSNs. However, little is presently known regarding early spine development in MSNs occurring before the arrival of cortical inputs and in particular about the role of DA and D1 (D1R) and D2 (D2R) DA receptors. A cell culture model reconstituting early cellular interactions between MSNs, intrinsic cholinergic interneurons and DA neurons was used to study the role of DA in spine formation. After 5 or 10 days in vitro, the presence of DA neurons increased the number of immature spine-like protrusions. In MSN monocultures, chronic activation of D1R or D2R also increased the number of spines and spinophilin expression in MSNs, suggesting a direct role for these receptors. In DA-MSN cocultures, chronic blockade of D1R or D2R reduced the number of dendritic spines. Interestingly, the combined activation or blockade of both D1R and D2R failed to elicit more extensive spine formation, suggesting that both receptors act through a mechanism that is not additive. Finally, we found increased ionotropic glutamate receptor responsiveness and miniature excitatory postsynaptic current (EPSC) frequency in DA-MSN co-cultures, in parallel with a higher number of spines containing PSD-95, suggesting that the newly formed spines present functional post-synaptic machinery preparing the MSNs to receive additional glutamatergic contacts. These results represent a first step in the understanding of how dopamine neurons promote the structural plasticity of MSNs during the development of basal ganglia circuits.

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

    Science.gov (United States)

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

    2015-02-01

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

  12. Ammonium sulphate precipitation of recombinant adenovirus from culture medium: an easy method to increase the total virus yield.

    Science.gov (United States)

    Schagen, F H; Rademaker, H J; Rabelink, M J; van Ormondt, H; Fallaux, F J; van der Eb, A J; Hoeben, R C

    2000-09-01

    In the majority of the methods for purifying and concentrating recombinant adenoviruses (rAds) the virus that is associated with the helper cells is harvested, while the virus that is present in the cell-culture medium is discarded. During routine propagation of adenovirus type-5 vectors at optimised conditions we noted that, on average, 47% of the total amount of virus is present in the culture medium. To recover and concentrate these rAds from the medium, we devised a method, which is based on ammonium sulphate ((NH4)2SO4) precipitation. At 40% (NH4)2SO4 saturation, 95 +/- 6% of the available virus precipitates from the medium, while the majority of the protein (85%) remains in solution. In contrast to adenovirus precipitation with polyethylene glycol, the (NH4)2SO4 precipitation technique allows collection of precipitated rAds by filtration. We demonstrate here that (NH4)2SO4 precipitation of rAds from cell-culture medium is a simple and fast technique that can be used in combination with standard virus isolation methods to increase the yields of rAds.

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

    Directory of Open Access Journals (Sweden)

    Szymańska Katarzyna

    2016-05-01

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

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

    Directory of Open Access Journals (Sweden)

    Manju Pathak

    2012-10-01

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

  15. 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

    2004-01-01

    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.

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

    Directory of Open Access Journals (Sweden)

    Krzysztof Makowski

    2017-07-01

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

  17. Growth and development of rabbit oocytes in vitro: effect of fetal bovine serum concentration on culture medium.

    Science.gov (United States)

    Sugimoto, H; Kida, Y; Miyamoto, Y; Kitada, K; Matsumoto, K; Saeki, K; Taniguchi, T; Hosoi, Y

    2012-09-15

    The objective was to develop a culture system that produced blastocyst stage embryos from rabbit oocytes grown in vitro. Two experiments were performed. First, various concentrations of fetal bovine serum (FBS, 0, 0.05, 0.5 and 5%) were used in the culture medium for in vitro growth (IVG) of oocytes recovered from follicles 200 to 299 μm in diameter. Intracytoplasmic sperm injection (ICSI) was performed on mature oocytes obtained after IVG for 8 days and in vitro maturation for 14 to 16 h. Rates of survival and pronuclear formation after ICSI were higher for oocytes grown in a medium with 0.05% FBS compared to oocytes grown in a medium lacking FBS (97.6 vs. 76.9%, 97.5 vs. 70%, P cultured in 0.05% FBS, oxygen consumption and the number of cells were analyzed. Blastocysts from oocytes grown in vitro with 0.05% FBS had reduced oxygen consumption and number of cells compared with those from ovulated oocytes (21.66 ± 4.54 × 10(14) vs. 50.19 ± 4.61 × 10(14) mol/sec, 244 ± 25 vs. 398 ± 24, P vitro with 0.05% FBS achieved pregnancy, but pregnancies were not maintained to term. In conclusion, the addition of 0.05% FBS to the culture medium for IVG improved developmental competence of rabbit oocytes grown in vitro.

  18. Structure of polysaccharides from mycelium and culture medium of Phellinus nigricans using submerged fermentation

    Institute of Scientific and Technical Information of China (English)

    LI Xia; JIAO LiLi; ZHANG Xu; TIAN WenMin; CHEN Shan; ZHANG LiPing

    2008-01-01

    Two water-soluble polysaccharides, PNW1 and PNM1, were respectively isolated from the mycelium and its culture medium of Phellinus nigricans using submerged fermentation before determining their effects on inhibiting the growth of transplantable tumors in mice. The results of the pharmaceutical experiments showed that oral administration of PNW1 and PNM1 (at a dose of 400 mg/kg) inhibited the growth of tumor of mice-transplanted Sarcoma 180 in vivo. Moreover, a higher inhibition ratio of PNW1 (74.70%) was obtained as compared with PNM1 (55.84%). The averaged molecular weight of PNW1 and PNM1 was determined to be 33 and 29 kD, respectively. Both PNW1 and PNM1 were consisted of glu-cose, galactose, mannose, arabinose and fucose. The major structural features of PNW1 and PNM1 were elucidated using partial acid hydrolysis, periodate oxidation, Smith degradation,13C-NMR, me-thylation and GC-MS. On the basis of these results, the repeating units of PNW1 and PNM1 were estab-lished.

  19. Structure of polysaccharides from mycelium and culture medium of Phellinus nigricans using submerged fermentation

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Two water-soluble polysaccharides, PNW1 and PNM1, were respectively isolated from the mycelium and its culture medium of Phellinus nigricans using submerged fermentation before determining their effects on inhibiting the growth of transplantable tumors in mice. The results of the pharmaceutical experiments showed that oral administration of PNW1 and PNM1 (at a dose of 400 mg/kg) inhibited the growth of tumor of mice-transplanted Sarcoma 180 in vivo. Moreover, a higher inhibition ratio of PNW1 (74.70%) was obtained as compared with PNM1 (55.84%). The averaged molecular weight of PNW1 and PNM1 was determined to be 33 and 29 kD, respectively. Both PNW1 and PNM1 were consisted of glu- cose, galactose, mannose, arabinose and fucose. The major structural features of PNW1 and PNM1 were elucidated using partial acid hydrolysis, periodate oxidation, Smith degradation, 13C-NMR, me- thylation and GC-MS. On the basis of these results, the repeating units of PNW1 and PNM1 were estab- lished.

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

    Directory of Open Access Journals (Sweden)

    Małgorzata Cackowska

    2011-11-01

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

  1. Effects of cultural medium on the formation and antitumor activity of polysaccharides by Cordyceps gunnii.

    Science.gov (United States)

    Zhu, Zhen-Yuan; Liu, Xiao-Cui; Tang, Ya-Li; Dong, Feng-Ying; Sun, Hui-Qing; Chen, Lu; Zhang, Yong-Min

    2016-10-01

    The effects of culture medium composition (i.e., carbon and nitrogen sources) on the growth of mycelia, molecular weight distribution and antitumor activity of intracellular polysaccharides (IPS) from Cordyceps gunnii were investigated. Sucrose and peptone were proved to be the best carbon and nitrogen sources for mycelia growth and remarkably improved IPS production. When the sucrose concentration was 2.0%, the mycelium yield reached up to 15.94±1.26 g/L, but with lower IPS yield; whereas the sucrose concentration was 4.5%, IPS yield reached to a maximum of 138.78±3.89 mg/100 mL. The effects of different carbon/nitrogen (C/N) ratios with equal amounts of carbon source matter on the mycelia and IPS formation were optimized. It found that the yield of mycelia and IPS were both reached to the highest at a C/N ratio of 10:3. In addition, the IPS had the highest macro molecular polysaccharide content and antitumor activity when sucrose concentration was 3.5% and the C/N ratio was 10:1.5. Thus, there was a positive correlation between molecular weight distribution and antitumor activity of IPS by C. gunnii. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  2. In vitro culture of Babesia bovis in a bovine serum-free culture medium supplemented with insulin, transferrin, and selenite.

    Science.gov (United States)

    Rojas Martínez, C; Rodríguez-Vivas, R I; Figueroa Millán, J V; Acosta Viana, K Y; Gutiérrez Ruiz, E J; Álvarez Martínez, J A

    2016-11-01

    Bovine serum is an important factor for the optimal growth of Babesia bovis in vitro. This protozoan can be cultured in M-199 with Earle's salts medium (M-199) supplemented with 40% bovine serum (BS). In the present study, four media were assessed along with the control medium M-199. The effect on the proliferation of B. bovis in vitro was tested when these media were combined with insulin (Ins), transferrin (Trans) and selenite (Sel) in the absence of bovine serum. Treatment with Advanced DMEM/F12 medium (A-DMEM/F12) achieved the highest percentage of parasitized erythrocytes (PPE), reaching a maximum value of 9.59%. A-DMEM/F12 medium supplemented with a mixture of Ins (2000 mg/L), Trans (1100 mg/L), and Sel (1.34 mg/L) allowed for the adaptation and proliferation of B. bovis without bovine serum, showed a constant increase in PPE, and reached a maximum value of 9.7% during seven cycles of in vitro culture. It was concluded that continuous proliferation of B. bovis in vitro could be achieved using A-DMEM/F12 medium supplemented with Ins-Trans-Sel, without bovine serum. After adaptation for proliferation in serum-free medium, the B. bovis strain of parasites could have future use in the study of this economically important protozoan species that affects cattle.

  3. Low-serum culture with novel medium promotes maxillary/mandibular bone marrow stromal cell proliferation and osteogenic differentiation ability.

    Science.gov (United States)

    Suehiro, Fumio; Ishii, Masakazu; Asahina, Izumi; Murata, Hiroshi; Nishimura, Masahiro

    2017-02-16

    The purpose of this study was to evaluate the effect of low-serum STK2 medium on the isolation and osteogenic differentiation of human maxillary/mandibular bone marrow stromal cells (MBMSCs). Human MBMSCs were obtained from patients undergoing dental implant treatment. These cells were cultured in serum-free medium or STK2 medium containing 1  % fetal bovine serum (low-serum) or α-MEM containing 10  % fetal bovine serum (control). Proliferation on the culture surface, cell surface antigen expression, and mRNA levels of neural crest and osteogenic markers were examined. Alkaline phosphatase assay and Alizarin red staining were used to assess osteogenic differentiation potential. Immunoblotting analysis was performed to detect ERK phosphorylation. Low-serum and control MBMSCs were positive for CD73, CD90, and CD105 and negative for CD14, CD34, CD45, CD271, and HLA-DR. CD140a was absent in low-serum cells but present in control cells. Low-serum MBMSCs proliferated more than control MBMSCs. After induction of osteogenic differentiation, alkaline phosphatase activity and osteocalcin mRNA levels were higher in low-serum MBMSCs than in control cells, and Alizarin red staining was stronger in low-serum MBMSCs than in control cells. Low-serum culture promoted ERK phosphorylation. MBMSCs precultured in low-serum medium exhibited a greater cumulative cell number and a higher osteogenic differentiation capacity than those cultured in control medium. These findings indicate that low-serum STK2 culture might be useful to promote MBMSC proliferation and osteogenic differentiation. This method requires less autologous blood collection for cell expansion than conventional methods, thus reducing the burden on patients.

  4. THE USE OF CONTINUOUS, TEMPORARY IMMERSION BIOREACTOR SYSTEM AND SEMISOLID CULTURE MEDIUM FOR THE PRODUCTION OF Eucalyptus camaldulensis CLONES

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    Evânia Galvão Mendonça

    2016-01-01

    Full Text Available The plant micro-propagation in bioreactor systems is regarded as one way to reduce cost by automation and production scheduling. This research was carried out in order to obtain an efficient procedure for clone production of Eucalyptus camaldulensis on different types of bioreactor including continuous and temporary immersion bioreactor. To do so, the apical meristems (1 mm and the apical meristems with adjacent tissue (2,5 mm were used as initial explants. These tissues were cultured, for 60 days, in semisolid culture medium supplemented with 1 mg L -1 indole acetic acid (IAA and 0.32 mg L -1 benzylaminopurine (BA. After 60 days, the meristems with adjacent tissue were transferred to a continuous immersion bioreactor and maintained in dark or light conditions. In order to verify the effect of the explant source on bioreactor multiplication, the explants subcultured from meristems multiplied in semisolid culture medium and the meristems multiplied in continuous immersion bioreactor were tested and maintained in dark conditions. After establishing this parameters, the multiplication experiments were carried out in continuous and temporary immersion and the multiplied explants were then rooted in MS medium supplemented with 0, 2, 4, 8 and 20 mg L -1 indole butyric acid (IBA and kept in the dark or under controlled lighting conditions. After that, the rooting the plants were acclimatized in mist chamber. The meristem with adjacent tissue favored a greater number of buds/explants. The continuous immersion bioreactor in the dark provided higher shoots number and multiplication rate. The rooting was better on culture medium without auxin and kept in the dark for 15 days or the culture medium supplemented with auxin and maintained under light with 100% plantlet rooting. The Eucalyptus camaldulensis acclimatization was efficient, with high survival rate (76%. It was possible to establish the procedure for bioreactor micro-propagation of Eucalyptus

  5. Cell death in a co-culture of hepatocellular carcinoma cells and human umbilical vascular endothelial cells in a medium lacking glucose and arginine.

    Science.gov (United States)

    Tomizawa, Minoru; Shinozaki, Fuminobu; Motoyoshi, Yasufumi; Sugiyama, Takao; Yamamoto, Shigenori; Ishige, Naoki

    2017-01-01

    Human primary hepatocytes are able to survive in a medium without glucose and arginine that is instead supplemented with galactose and ornithine (hepatocyte selection medium; HSM). This is because the cells produce glucose and arginine by the action of galactokinase (GALK) and ornithine carbamoyltransferase (OTC), respectively. It was expected that hepatocellular carcinoma (HCC) cells do not survive in HSM. In the current study, HCC cell lines (namely HLE, HLF, PLC/PRL/5, Hep3B and HepG2) and human umbilical vascular endothelial cells (HUVECs) were cultured in HSM, and the expression levels of GALK1, GALK2 and OTC were analyzed by reverse transcription-quantitative polymerase chain reaction. HLE, HLF and PLC/PRL/5 cells died on day 11, while Hep3B, HepG2 and HUVECs died on day 7. HLF cells were further analyzed as these cells had lower expression levels of GALK1, GALK2 and OTC compared with adult liver cells, and survived until day 11. In these cells, the expression levels of GALK1, GALK2 and OTC did not change on days 3 and 7 as compared to day 0. In addition, a co-culture of HLF cells with HUVECs was established and the medium was changed to HSM. It was observed that HLF cells and HUVECs in co-culture were damaged in HSM. In summary, HCC cells and HUVECs died in a medium without glucose and arginine that was supplemented with galactose and ornithine. HCC cells and HUVECs were damaged in HSM, suggesting a potential application for treatment with the medium.

  6. What Kind of Signaling Maintains Pluripotency and Viability in Human-Induced Pluripotent Stem Cells Cultured on Laminin-511 with Serum-Free Medium?

    Science.gov (United States)

    Nakashima, Yoshiki; Omasa, Takeshi

    2016-01-01

    Xeno-free medium contains no animal-derived components, but is composed of minimal growth factors and is serum free; the medium may be supplemented with insulin, transferrin, and selenium (ITS medium). Serum-free and xeno-free culture of human-induced pluripotent stem cells (hiPSCs) uses a variety of components based on ITS medium and Dulbecco's modified Eagle's medium/Ham's nutrient mixture F12 (DMEM/F12) that contain high levels of iron salt and glucose. Culture of hiPSCs also requires scaffolding materials, such as extracellular matrix, collagen, fibronectin, laminin, proteoglycan, and vitronectin. The scaffolding component laminin-511, which is composed of α5, β1, and γ1 chains, binds to α3β1, α6β1, and α6β4 integrins on the cell membrane to induce activation of the PI3K/AKT- and Ras/MAPK-dependent signaling pathways. In hiPSCs, the interaction of laminin-511/α6β1 integrin with the cell-cell adhesion molecule E-cadherin confers protection against apoptosis through the Ras homolog gene family member A (RhoA)/Rho kinase (ROCK) signaling pathway (the major pathways for cell death) and the proto-oncogene tyrosine-protein kinase Fyn (Fyn)-RhoA-ROCK signaling pathway. The expression levels of α6β1 integrin and E-cadherin on cell membranes are controlled through the activation of insulin receptor/insulin, FGF receptor/FGF2, or activin-like kinase 5 (ALK5)-dependent TGF-β signaling. A combination of growth factors, medium constituents, cell membrane-located E-cadherin, and α6β1 integrin-induced signaling is required for pluripotent cell proliferation and for optimal cell survival on a laminin-511 scaffold. In this review, we discuss and explore the influence of growth factors on the cadherin and integrin signaling pathways in serum-free and xeno-free cultures of hiPSCs during the preparation of products for regenerative medicinal therapies. In addition, we suggest the optimum serum-free medium components for use with laminin-511, a new scaffold for hi

  7. A stochastic model of gene-culture coevolution suggested by the "culture historical hypothesis" for the evolution of adult lactose absorption in humans.

    Science.gov (United States)

    Aoki, K

    1986-05-01

    A stochastic model of gene-culture coevolution, suggested by the "culture historical hypothesis" of Simoons and McCracken, is presented. According to this hypothesis, adult lactose absorption, believed to be an autosomal dominant trait, attained a high frequency in some human populations due to the positive selection pressure induced by culturally determined milk use in those populations. Two-dimensional Kolmogorov backward equations with appropriate boundary conditions are derived for the ultimate fixation probability of milk users, of the gene for adult lactose absorption, and of both jointly, and for the average time until fixation of the gene. These boundary value problems are solved numerically by the Gauss-Seidel method. I define a theoretical measure of the correlation between gene and culture in terms of the three ultimate fixation probabilities. Monte Carlo simulations are conducted to check and extend the numerical results and also to obtain the first arrival time at gene frequency 0.70, which is approximately the highest observed frequency in any population. Two results that pertain to the culture historical hypothesis are obtained. First, the incomplete correlation observed between adult lactose absorption and milk use does not necessarily constitute evidence against the hypothesis. Second, for the postulated genetic change to have occurred within the 6000-year period since the advent of dairying, either the effective population size was of the order of 100, or, if it was of larger order, the selection coefficient probably had to exceed 5%.

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

    Directory of Open Access Journals (Sweden)

    Rauquírio André Albuquerque Marinho da Costa

    1999-01-01

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

  9. Evaluation of Bacillus sphaericus bioinsecticide produced with white soybean meal as culture medium for the control of Culex (Culex) quinquefasciatus.

    Science.gov (United States)

    Melo, André L A; Soccol, Carlos R; Thomaz-Soccol, Vanete; Nogueira, Miodeli

    2009-03-01

    Bioinsecticides are shown to be useful in control programs to prevent several diseases, based on their specificity and efficiency against insect vectors. In the current study a bioinsecticide based on Bacillus sphaericus was produced using a white soybean culture medium and applied to larvae of Culex quinquefasciatus, the susceptible species, and Aedes aegypti, the refractory species used as the negative control. Efficacy was compared with that of the product fermented with the Luria Bertani (LB) reference medium. The experiments showed that C. quinquefasciatus was highly susceptible to the product prepared with white soybean meal, reaching 100% larval mortality even at 10mg/L, while A. aegypti failed to reach 70% mortality at a concentration of 1g/L. By comparison with the reference medium, the proposed culture medium showed high larvicidal power, reaching a LD90 of 2.26 mg/L, while 4.37 mg/L was needed for the LB medium to achieve the same mortality rate. Cost comparison between the formulations favored the use of the bioinsecticide produced with white soybean meal. After factoring in the LD90 value, the cost ratio favored the new raw material by nearly 1:220.

  10. Elongation factor 1-alpha is released into the culture medium during growth of Giardia intestinalis trophozoites.

    Science.gov (United States)

    Skarin, Hanna; Ringqvist, Emma; Hellman, Ulf; Svärd, Staffan G

    2011-04-01

    The molecular pathogenesis of the intestinal parasite Giardia intestinalis is still not fully understood but excretory-secretory products have been suggested to be important during host-parasite interactions. Here we used SDS-PAGE gels and MALDI-TOF analysis to identify proteins released by Giardia trophozoites during in vitro growth. Serum proteins (mainly bovine serum albumin) in the growth medium, bind to the parasite surface and they are continuously released, which interfere with parasite secretome characterization. However, we identified two released Giardia proteins: elongation factor-1 alpha (EF-1α) and a 58 kDa protein, identified as arginine deiminase (ADI). This is the first description of EF-1α as a released/secreted Giardia protein, whereas ADI has been identified in an earlier secretome study. Two genes encoding EF-1α were detected in the Giardia WB genome 35 kbp apart with almost identical coding sequences but with different promoter and 3' regions. Promoter luciferase-fusions showed that both genes are transcribed in trophozoites. The EF-1α protein localizes to the nuclear region in trophozoites but it relocalizes to the cytoplasm during host-cell interaction. Recombinant EF-1α is recognized by serum from giardiasis patients. Our results suggest that released EF-1α protein can be important during Giardia infections.

  11. Stem cells from human exfoliated deciduous teeth differentiate toward neural cells in a medium dynamically cultured with Schwann cells in a series of polydimethylsiloxanes scaffolds

    Science.gov (United States)

    Su, Wen-Ta; Pan, Yu-Jing

    2016-08-01

    Objective. Schwann cells (SCs) are primary structural and functional cells in the peripheral nervous system. These cells play a crucial role in peripheral nerve regeneration by releasing neurotrophic factors. This study evaluated the neural differentiation potential effects of stem cells from human exfoliated deciduous teeth (SHEDs) in a rat Schwann cell (RSC) culture medium. Approach. SHEDs and RSCs were individually cultured on a polydimethylsiloxane (PDMS) scaffold, and the effects of the RSC medium on the SHEDs differentiation between static and dynamic cultures were compared. Main results. Results demonstrated that the SHED cells differentiated by the RSC cultured medium in the static culture formed neurospheres after 7 days at the earliest, and SHED cells formed neurospheres within 3 days in the dynamic culture. These results confirm that the RSC culture medium can induce neurospheres formation, the speed of formation and the number of neurospheres (19.16 folds high) in a dynamic culture was superior to the static culture for 3 days culture. The SHED-derived spheres were further incubated in the RSCs culture medium, these neurospheres continuously differentiated into neurons and neuroglial cells. Immunofluorescent staining and RT-PCR revealed nestin, β-III tubulin, GFAP, and γ-enolase of neural markers on the differentiated cells. Significance. These results indicated that the RSC culture medium can induce the neural differentiation of SHED cells, and can be used as a new therapeutic tool to repair nerve damage.

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

    Directory of Open Access Journals (Sweden)

    José Soares do Nascimento

    2007-07-01

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

  13. Optimization of culture medium for lactosucrose ( G-beta-D-galactosylsucrose) Production by Sterigmatomyces elviae mutant using statistical analysis.

    Science.gov (United States)

    Lee, Jong Ho; Lim, Jung Soo; Song, Yoon Seok; Kang, Seong Woo; Park, Chulhwan; Kim, Seung Wook

    2007-12-01

    In this study, the optimization of culture medium using a Sterigmatomyces elviae mutant was investigated using statistical analysis to increase the cell mass and lactosucrose ((4)G-beta-D-galactosylsucrose) production. In basal medium, the cell mass and lactosucrose production were 4.12 g/l and 140.91 g/l, respectively. However, because of the low cell mass and lactosucrose production, optimization of culture medium was carried out to increase the cell mass and lactosucrose production. Culture media were optimized by the S. elviae mutant using analysis of variance (ANOVA) and response surface methodology (RSM). Central composite designs using RSM were utilized in this investigation. Quadratic models were obtained for cell mass and lactosucrose production. In the case of cell mass, optimal components of the medium were as follows: sucrose 1.13%, yeast extract 0.99%, bactopeptone 2.96%, and ammonium sulfate 0.40%. The predicted maximum value of cell mass was about 5.20 g/l and its experimental value was 5.08 g/l. In the case of lactosucrose production, optimal components of the medium were as follows: sucrose 0.96%, yeast extract 1.2%, bactopeptone 3.0%, and ammonium sulfate 0.48%. Then, the predicted maximum value of lactosucrose production was about 194.12 g/l and the corresponding experimental value was about 183.78 g/l. Therefore, by culturing using predicted conditions, the real cell mass and lactosucrose production increased to 23.3% and 30.42%, respectively.

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

    Directory of Open Access Journals (Sweden)

    Endang Triwulaninngsih

    2002-03-01

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

  15. Detection and characterization of silver nanoparticles and dissolved species of silver in culture medium and cells by AsFlFFF-UV-Vis-ICPMS: application to nanotoxicity tests.

    Science.gov (United States)

    Bolea, E; Jiménez-Lamana, J; Laborda, F; Abad-Álvaro, I; Bladé, C; Arola, L; Castillo, J R

    2014-03-07

    A methodology based on Asymmetric Flow Field-Flow Fractionation (AsFlFFF) coupled with UV-Vis absorption spectrometry and ICP mass spectrometry (ICPMS) has been developed and applied to the study of silver nanoparticles (AgNPs) and dissolved species of silver in culture media and cells used in cytotoxicity tests. The effect of a nano-silver based product (protein stabilized silver nanoparticles ca. 15 nm average diameter) on human hepatoma (HepG2) cell viability has been studied. UV-Vis absorption spectrometry provided information about the nature (organic vs. nanoparticle) of the eluted species, whereas the silver was monitored by ICPMS. A shift towards larger hydrodynamic diameters was observed in the AgNPs after a 24 hour incubation period in the culture medium, which suggests a "protein corona" effect. Silver(I) associated with proteins present in the culture medium has also been detected, as a consequence of the oxidation process experimented by the AgNPs. However, the Ag(I) released into the culture medium did not justify the toxicity levels observed. AgNPs associated with the cultured HepG2 cells were also identified by AsFlFFF, after applying a solubilisation process based on the use of tetramethylammonium hydroxide (TMAH) and Triton X-100. These results have been confirmed by transmission electronic microscopy (TEM) analysis of the fractions collected from the AsFlFFF. The effect of AgNPs on HepG2 cells has been compared to that caused by silver(I) as AgNO3 under the same conditions. The determination of the total content of silver in the cells confirms that a much larger mass of silver as AgNPs with respect to AgNO3 (16 to 1) is needed to observe a similar toxicity.

  16. Increased diazinon hydrolysis to 2-isopropyl-6-methyl-4-pyrimidinol in liquid medium by a specific Streptomyces mixed culture.

    Science.gov (United States)

    Briceño, G; Schalchli, H; Rubilar, O; Tortella, G R; Mutis, A; Benimeli, C S; Palma, G; Diez, M C

    2016-08-01

    Actinobacteria identified as Streptomyces spp. were evaluated for their ability to remove diazinon as the only carbon source from a liquid medium. Single cultures of Streptomyces strains were exposed to diazinon at a concentration of 50 mg L(-1). After 96 h incubation, six of the eight cultures grew and five strains showed an increase in their total protein concentrations and changes in their protein profile. Up to 32% of the diazinon was removed by the single Streptomyces cultures. A compatibility assay showed that the different Streptomyces species were not antagonistic. Twenty-six mixed cultures were then prepared. Diazinon removal was increased when mixed cultures were used, and maximum diazinon removal of 62% was observed when the Streptomyces spp. strains AC5, AC9, GA11 and ISP13 were mixed; this was defined as the selected mixed culture (SMC). Diazinon removal was positively influenced by the addition of glucose into the liquid medium. Our study showed a diazinon degradation rate of 0.025 h(-1), half-life of 28 h(-1) and 2-isopropyl-6-methyl-4-pyrimidinol (IMHP) production of 0.143 mg L h(-1). Rapid diazinon hydrolysis to IMHP was associated with a decrease in the pH of the medium as a consequence of microbial glucose metabolism and organic acid exudation. Moreover, the SMC of Streptomyces was able to remove IMHP. This work constitutes a new, if not the only, report on diazinon degradation by mixed cultures of Streptomyces spp. Given the high levels of diazinon removal, the SMC formed by four Streptomyces strains has the potential to be used to treat the diazinon present in environmental matrices.

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

    DEFF Research Database (Denmark)

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

    2013-01-01

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

  18. Chemical properties and antioxidant activity of exopolysaccharides fractions from mycelial culture of Inonotus obliquus in a ground corn stover medium.

    Science.gov (United States)

    Xiang, Yuling; Xu, Xiangqun; Li, Juan

    2012-10-15

    The medicinal mushroom Inonotus obliquus has been a folk remedy for a long time in East-European and Asian countries. We first reported the enhancement in production and antioxidant activity of exopolysaccharides by I. obliquus culture under lignocellulose decomposition. In this study, the two different sources of exopolysaccharides from the control medium and the lignocellulose (corn stover) containing medium by I. obliquus in submerged fermentation were fractionated and purified by chromatography. The exopolysaccharides from the corn stover-containing medium presented significantly stronger hydroxyl and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity than the control. Three fractions from the control medium and the corn stover-containing medium were isolated respectively. The fraction of DEPL3 from the corn stover-containing medium with the highest protein content (38.3%), mannose content (49.6%), and the lowest molecular weight (29 kDa) had the highest antioxidant activity with the lowest IC50 values. In conclusion, lignocellulose decomposition changed the chemical characterisation and significantly enhanced the antioxidant activity of exopolysaccharide fractions.

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

    Science.gov (United States)

    Henríquez, Mirtha; González, Ernesto; Marshall, Sergio H; Henríquez, Vitalia; Gómez, Fernando A; Martínez, Irene; Altamirano, Claudia

    2013-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Mirtha Henríquez

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

  1. Population structure of mixed Mycobacterium tuberculosis infection is strain genotype and culture medium dependent.

    Directory of Open Access Journals (Sweden)

    Madeleine Hanekom

    Full Text Available BACKGROUND: Molecular genotyping methods have shown infection with more than one Mycobacterium tuberculosis strain genotype in a single sputum culture, indicating mixed infection. AIM: This study aimed to develop a PCR-based genotyping tool to determine the population structure of M. tuberculosis strain genotypes in primary Mycobacterial Growth Indicator Tubes (MGIT and Löwenstein-Jensen (LJ cultures to identify mixed infections and to establish whether the growth media influenced the recovery of certain strain genotypes. METHOD: A convenience sample of 206 paired MGIT and LJ M. tuberculosis cultures from pulmonary tuberculosis patients resident in Khayelitsha, South Africa were genotyped using an in-house PCR-based method to detect defined M. tuberculosis strain genotypes. RESULTS: The sensitivity and specificity of the PCR-based method for detecting Beijing, Haarlem, S-family, and LAM genotypes was 100%, and 75% and 50% for detecting the Low Copy Clade, respectively. Thirty-one (15% of the 206 cases showed the presence of more than one M. tuberculosis strain genotype. Strains of the Beijing and Haarlem genotypes were significantly more associated with a mixed infection (on both media when compared to infections with a single strain (Beijing MGIT p = 0.02; LJ, p<0.01 and (Haarlem: MGIT p<0.01; LJ, p = 0.01. Strains with the Beijing genotype were less likely to be with "other genotype" strains (p<0.01 while LAM, Haarlem, S-family and LCC occurred independently with the Beijing genotype. CONCLUSION: The PCR-based method was able to identify mixed infection in at least 15% of the cases. LJ media was more sensitive in detecting mixed infections than MGIT media, implying that the growth characteristics of M. tuberculosis on different media may influence our ability to detect mixed infections. The Beijing and Haarlem genotypes were more likely to occur in a mixed infection than any of the other genotypes tested suggesting pathogen

  2. Supplementation of bovine embryo culture medium with L-arginine improves embryo quality via nitric oxide production.

    Science.gov (United States)

    Santana, Priscila Di Paula Bessa; Silva, Thiago Velasco Guimarães; da Costa, Nathália Nogueira; da Silva, Bruno Barauna; Carter, Timothy Frederick; Cordeiro, Marcela da Silva; da Silva, Bruno José Martins; Santos, Simone do Socorro Damasceno; Herculano, Anderson Manoel; Adona, Paulo Roberto; Ohashi, Otávio Mitio; Miranda, Moysés dos Santos

    2014-10-01

    Nitric oxide (NO) is a cell-signaling molecule that regulates a variety of molecular pathways. We investigated the role of NO during preimplantation embryonic development by blocking its production with an inhibitor or supplementing in vitro bovine embryo cultures with its natural precursor, L-arginine, over different periods. Endpoints evaluated included blastocyst rates, development kinetics, and embryo quality. Supplementation with the NO synthase inhibitor N-Nitro-L-arginine-methyl ester (L-NAME) from Days 1 to 8 of culture decreased blastocyst (P quality (P culture period positively correlated with the increased embryo hatching rates and quality (P culture medium with L-arginine favors preimplantation development of bovine embryos. © 2014 Wiley Periodicals, Inc.

  3. Bioleaching of sphalerite by Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans cultured in 9K medium modified with pyrrhotite

    Institute of Scientific and Technical Information of China (English)

    CHEN Song; QIU Guan-zhou; QIN Wen-qing; LAN Zhuo-yue

    2008-01-01

    Elective culture of Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans in 9K medium modified with pyrrhotite was studied. Bioleaching of flotation concentrate of sphalerite by the selected bacteria was carried out. The results show that the microorganisms cultured by pyrrhotite are a mixture of Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans, of which the capability to oxidize ferrous to ferric irons is enhanced by the high mass ratio of Fe to S in pyrrhotite. Three pyrrhotite samples were separated into various parts with corresponding S/Fe ratios by magnetic separation and were used to culture the elective bacteria as the substrate. The association of the cultures could provide a more rapid and complete oxidation of sphalerite than that of bacteria cultivated by conventional methods.

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

    DEFF Research Database (Denmark)

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

    2017-01-01

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

  5. Controlled clinical comparison of plastic versus glass bottles of BacT/ALERT PF medium for culturing blood from children.

    Science.gov (United States)

    Petti, Cathy A; Mirrett, Stanley; Woods, Christopher W; Reller, L Barth

    2005-01-01

    The plastic pediatric BacT/ALERT (bioMérieux, Durham, N.C.) PF (PPF) is a new nonvented aerobic culture medium in a clear plastic bottle designed to prevent breakage. We compared the performance of the new PPF bottle to that of the present glass BacT/ALERT PF bottle for the recovery of microorganisms as well as for the time to detection of growth in samples of blood obtained for culture from children. We found that the PPF and PF bottles were comparable for recovery of microorganisms and that the safety advantage of plastic bottles can be achieved without compromising performance.

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

    DEFF Research Database (Denmark)

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

    2016-01-01

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

  7. Partial purification and characterization of an endo-alpha-N-acetylgalactosaminidase from the culture medium of Streptomyces sp. OH-11242.

    OpenAIRE

    Ishii-Karakasa, I; Iwase, H.; Hotta, K; Tanaka, Y.; Omura, S

    1992-01-01

    For the purification of a new type of endo-alpha-N-acetylgalactosaminidase from the culture medium of Streptomyces sp. OH-11242 (endo-GalNAc-ase-S) [Iwase, Ishii, Ishihara, Tanaka, Omura & Hotta (1988) Biochem. Biophys. Res. Commun. 151, 422-428], a method for assaying enzyme activity was established. Using purified pig gastric mucus glycoprotein (PGM) as the substrate, oligosaccharides liberated from PGM were pyridylaminated, and the reducing terminal sugars of oligosaccharides larger than G...

  8. Medium Renewal Blocks Anti-Proliferative Effects of Metformin in Cultured MDA-MB-231 Breast Cancer Cells.

    Science.gov (United States)

    Rajh, Maruša; Dolinar, Klemen; Miš, Katarina; Pavlin, Mojca; Pirkmajer, Sergej

    2016-01-01

    Epidemiological studies indicate that metformin, a widely used type 2 diabetes drug, might reduce breast cancer risk and mortality in patients with type 2 diabetes. Metformin might protect against breast cancer indirectly by ameliorating systemic glucose homeostasis. Alternatively, it might target breast cancer cells directly. However, experiments using MDA-MB-231 cells, a standard in vitro breast cancer model, produced inconsistent results regarding effectiveness of metformin as a direct anti-cancer agent. Metformin treatments in cultured MDA-MB-231 cells are usually performed for 48-96 hours, but protocols describing renewal of cell culture medium during these prolonged treatments are rarely reported. We determined whether medium renewal protocol might alter sensitivity of MDA-MB-231 cells treated with metformin. Using the MTS assay, BrdU incorporation and Hoechst staining we found that treatment with metformin for 48-72 hours failed to suppress viability and proliferation of MDA-MB-231 cells if low-glucose (1 g/L) medium was renewed every 24 hours. Conversely, metformin suppressed their viability and proliferation if medium was not renewed. Without renewal glucose concentration in the medium was reduced to 0.1 g/L in 72 hours, which likely explains increased sensitivity to metformin under these conditions. We also examined whether 2-deoxy-D-glucose (2-DG) reduces resistance to metformin. In the presence of 2-DG metformin reduced viability and proliferation of MDA-MB-231 cells with or without medium renewal, thus demonstrating that 2-DG reduces their resistance to metformin. In sum, we show that medium renewal blocks anti-proliferative effects of metformin during prolonged treatments in low-glucose medium. Differences in medium renewal protocols during prolonged treatments might therefore lead to apparently inconsistent results as regards effectiveness of metformin as a direct anti-cancer agent. Finally, our results indicate that co-therapy with 2-DG and

  9. Medium Renewal Blocks Anti-Proliferative Effects of Metformin in Cultured MDA-MB-231 Breast Cancer Cells

    Science.gov (United States)

    Rajh, Maruša; Dolinar, Klemen; Miš, Katarina; Pavlin, Mojca; Pirkmajer, Sergej

    2016-01-01

    Epidemiological studies indicate that metformin, a widely used type 2 diabetes drug, might reduce breast cancer risk and mortality in patients with type 2 diabetes. Metformin might protect against breast cancer indirectly by ameliorating systemic glucose homeostasis. Alternatively, it might target breast cancer cells directly. However, experiments using MDA-MB-231 cells, a standard in vitro breast cancer model, produced inconsistent results regarding effectiveness of metformin as a direct anti-cancer agent. Metformin treatments in cultured MDA-MB-231 cells are usually performed for 48–96 hours, but protocols describing renewal of cell culture medium during these prolonged treatments are rarely reported. We determined whether medium renewal protocol might alter sensitivity of MDA-MB-231 cells treated with metformin. Using the MTS assay, BrdU incorporation and Hoechst staining we found that treatment with metformin for 48–72 hours failed to suppress viability and proliferation of MDA-MB-231 cells if low-glucose (1 g/L) medium was renewed every 24 hours. Conversely, metformin suppressed their viability and proliferation if medium was not renewed. Without renewal glucose concentration in the medium was reduced to 0.1 g/L in 72 hours, which likely explains increased sensitivity to metformin under these conditions. We also examined whether 2-deoxy-D-glucose (2-DG) reduces resistance to metformin. In the presence of 2-DG metformin reduced viability and proliferation of MDA-MB-231 cells with or without medium renewal, thus demonstrating that 2-DG reduces their resistance to metformin. In sum, we show that medium renewal blocks anti-proliferative effects of metformin during prolonged treatments in low-glucose medium. Differences in medium renewal protocols during prolonged treatments might therefore lead to apparently inconsistent results as regards effectiveness of metformin as a direct anti-cancer agent. Finally, our results indicate that co-therapy with 2-DG and

  10. Development of an optimized medium, strain and high-throughput culturing methods for Methylobacterium extorquens

    National Research Council Canada - National Science Library

    Delaney, Nigel F; Kaczmarek, Maria E; Ward, Lewis M; Swanson, Paige K; Lee, Ming-Chun; Marx, Christopher J

    2013-01-01

    .... Here we develop a new system for high-throughput batch culture of M. extorquens in microtiter plates by jointly optimizing the properties of the organism, the growth media and the culturing system...

  11. Concise Review: No Breakthroughs for Human Mesenchymal and Embryonic Stem Cell Culture: Conditioned Medium, Feeder Layer, or Feeder‐Free; Medium with Fetal Calf Serum, Human Serum, or Enriched Plasma; Serum‐Free, Serum Replacement Nonconditioned Medium, or Ad Hoc Formula? All That Glitters Is Not Gold

    National Research Council Canada - National Science Library

    Mannello, Ferdinando; Tonti, Gaetana A

    2007-01-01

    The choice of an optimal strategy of stem cell culture is at the moment an impossible task, and the elaboration of a culture medium adapted to the production of embryonic and adult mesenchymal stem...

  12. Pre-culturing of nodal explants in thidiazuron supplemented liquid medium improves in vitro shoot multiplication of Cassia angustifolia.

    Science.gov (United States)

    Siddique, I; Abdullwahab Bukhari, N; Perveen, K; Siddiqui, I; Anis, M

    2013-09-01

    An in vitro propagation system for Cassia angustifolia Vahl. has been developed. Due to the presence of sennosides, the demand of this plant has increased manyfold in global market. Multiple shoots were induced by culturing nodal explants excised from mature plants on a liquid Murashige and Skoog [8] medium supplemented with 5-100 μM of thidiazuron (TDZ) for different treatment duration (4, 8, 12 and 16 d). The optimal level of TDZ supplemented to the culture medium was 75 μM for 12 d induction period followed by subculturing in MS medium devoid of TDZ as it produced maximum regeneration frequency (87%), mean number of shoots (9.6 ± 0.33) and shoot length (4.4 ± 0.46 cm) per explant. A culture period longer than 12 d with TDZ resulted in the formation of fasciated or distorted shoots. Ex vitro rooting was achieved when the basal cut end of regenerated shoots was dipped in 200 μM indole-3-butyric acid (IBA) for half an hour followed by their transplantation in plastic pots filled with sterile soilrite where 85% plantlets grew well and all exhibited normal development. The present findings describe an efficient and rapid plant regeneration protocol that can further be used for genetic transformation studies.

  13. Complete transformation of ZnO and CuO nanoparticles in culture medium and lymphocyte cells during toxicity testing.

    Science.gov (United States)

    Ivask, Angela; Scheckel, Kirk G; Kapruwan, Pankaj; Stone, Vicki; Yin, Hong; Voelcker, Nicolas H; Lombi, Enzo

    2017-03-01

    Here, we present evidence on complete transformation of ZnO and CuO nanoparticles, which are among the most heavily studied metal oxide particles, during 24 h in vitro toxicological testing with human T-lymphocytes. Synchrotron radiation-based X-ray absorption near edge structure (XANES) spectroscopy results revealed that Zn speciation profiles of 30 nm and 80 nm ZnO nanoparticles, and ZnSO4- exposed cells were almost identical with the prevailing species being Zn-cysteine. This suggests that ZnO nanoparticles are rapidly transformed during a standard in vitro toxicological assay, and are sequestered intracellularly, analogously to soluble Zn. Complete transformation of ZnO in the test conditions was further supported by almost identical Zn spectra in medium to which ZnO nanoparticles or ZnSO4 was added. Likewise, Cu XANES spectra for CuO and CuSO4-exposed cells and cell culture media were similar. These results together with our observation on similar toxicological profiles of ZnO and soluble Zn, and CuO and soluble Cu, underline the importance of dissolution and subsequent transformation of ZnO and CuO nanoparticles during toxicological testing and provide evidence that the nano-specific effect of ZnO and CuO nanoparticles is negligible in this system. We strongly suggest to account for this aspect when interpreting the toxicological results of ZnO and CuO nanoparticles.

  14. Effect of nitrogen concentration in culture mediums on growth and enzyme production of Phanerochaete chrysosporium

    Institute of Scientific and Technical Information of China (English)

    GAO Da-wen; WEN Xiang-hua; QIAN Yi

    2005-01-01

    Effect of different nitrogen concentration in the mediums on growth and enzyme production of Phanerochaete chrysosporium was studied when glucose concentration was 10 g/L. The results showed that the medium contained 0.8 g/L ammonium tartrate is the best. It not only supply abundant nutrients for the growth of Phanerochaete chrysosporium, which make mycelia the best grow compared with the other medium, but also produce higher manganese-dependent peroxidase(Mnp) and laccase(Lac) activity. In addition, it is observed that the variation of mycelia surface is related to ligninolytic enzyme secreted by Phanerochaete chrysosporium. When the surface of mycelium pellets appeared burs, it predicts secondary metabolism begin. This experimentation demonstrated that when the ratio of carbon and nitrogen in nitrogen limited medium is equal to 100:8, growth and enzyme production of Phanerochaete chrysosporium is the best, it could achieve the maximum Mnp and Lac activity.

  15. Effect of nitrogen concentration in culture mediums on growth and enzyme production of Phanerochaete chrysosporium.

    Science.gov (United States)

    Gao, Da-wen; Wen, Xiang-hua; Qian, Yi

    2005-01-01

    Effect of different nitrogen concentration in the mediums on growth and enzyme production of Phanerochaete chrysosporium was studied when glucose concentration was 10 g/L. The results showed that the medium contained 0.8 g/L ammonium tartrate is the best. It not only supply abundant nutrients for the growth of Phanerochaete chrysosporium, which make mycelia the best grow compared with the other medium, but also produce higher manganese-dependent peroxidase (Mnp) and laccase (Lac) activity. In addition, it is observed that the variation of mycelia surface is related to ligninolytic enzyme secreted by Phanerochaete chrysosporium. When the surface of mycelium pellets appeared burs, it predicts secondary metabolism begin. This experimentation demonstrated that when the ratio of carbon and nitrogen in nitrogen limited medium is equal to 100:8, growth and enzyme production of Phanerochaete chrysosporium is the best, it could achieve the maximum Mnp and Lac activity.

  16. Studies on a chemically defined medium for in vitro culture of in vitro matured and fertilized porcine oocytes.

    Science.gov (United States)

    Iwasaki, T; Kimura, E; Totsukawa, K

    1999-03-01

    The present study evaluated the effects of various components in a chemically defined medium on the development of IVM/IVF porcine embryos. The investigated components included energy substrates (lactate, pyruvate or glucose, alone or in various combinations), amino acids (glutamine, glycine or alanine), PVP and HEPES buffer. The effects of each energy substrate were the same as the control. However, a mixture of lactate with either of the other energy substrates increased the development rate. Glutamine tended to decrease rate of the development more than other amino acids, and this inhibition was dose dependent. Both PVP and HEPES buffer did not affect development rate. However, more than 35 mM HEPES buffer induced fragmentation From the above results, a new culture medium was designed (supplemented with 0.276 mM glycine, 0.176 mM alanine, 15 mM HEPES buffer and 1% (wt/vol) PVP in BSA-free Whitten's medium with or without glucose). The new medium resulted in a higher embryo development rate (20.4 and 16.3%) than that obtained with the control medium (10.0%).

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

    Directory of Open Access Journals (Sweden)

    R. Gamaroff

    2013-02-01

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

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

    Directory of Open Access Journals (Sweden)

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

    2011-04-01

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

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

    Directory of Open Access Journals (Sweden)

    Ancasi-Espejo Ruth Gabriela

    2016-08-01

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

  20. Stirred tank bioreactor culture combined with serum-/xenogeneic-free culture medium enables an efficient expansion of umbilical cord-derived mesenchymal stem/stromal cells.

    Science.gov (United States)

    Mizukami, Amanda; Fernandes-Platzgummer, Ana; Carmelo, Joana G; Swiech, Kamilla; Covas, Dimas T; Cabral, Joaquim M S; da Silva, Cláudia L

    2016-08-01

    Mesenchymal stem/stromal cells (MSC) are being widely explored as promising candidates for cell-based therapies. Among the different human MSC origins exploited, umbilical cord represents an attractive and readily available source of MSC that involves a non-invasive collection procedure. In order to achieve relevant cell numbers of human MSC for clinical applications, it is crucial to develop scalable culture systems that allow bioprocess control and monitoring, combined with the use of serum/xenogeneic (xeno)-free culture media. In the present study, we firstly established a spinner flask culture system combining gelatin-based Cultispher(®) S microcarriers and xeno-free culture medium for the expansion of umbilical cord matrix (UCM)-derived MSC. This system enabled the production of 2.4 (±1.1) x10(5) cells/mL (n = 4) after 5 days of culture, corresponding to a 5.3 (±1.6)-fold increase in cell number. The established protocol was then implemented in a stirred-tank bioreactor (800 mL working volume) (n = 3) yielding 115 million cells after 4 days. Upon expansion under stirred conditions, cells retained their differentiation ability and immunomodulatory potential. The development of a scalable microcarrier-based stirred culture system, using xeno-free culture medium that suits the intrinsic features of UCM-derived MSC represents an important step towards a GMP compliant large-scale production platform for these promising cell therapy candidates. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

    Directory of Open Access Journals (Sweden)

    Yong-Hao Chen

    2012-12-01

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

  2. Effects of Medium Constituents on Growth and Canthinone Accumulation in Cell Suspension Cultures of Eurycoma longifolia Jack

    Directory of Open Access Journals (Sweden)

    LUTHFI AZIZ MAHMUD SIREGAR

    2009-06-01

    Full Text Available The effect of various macronutrients, micronutrients and sucrose on growth and canthinone alkaloid production in cell suspension cultures of Pasak Bumi (Eurycoma longifolia Jack was investigated. The optimum macronutrients and micronutrients content for the high alkaloid production of E. longifolia Jack was different to that found in the Murashige and Skoog (MS medium. The highest amount of alkaloids, 9-hydroxycanthin-6-one and 9-methoxycanthin-6-one, could be obtained from E. longifolia Jack cells cultured in modified MS liquid medium that containing macronutrients: 21.50 mM NH4NO3, 14.25 mM KNO3, 7.50 mM CaCl2•2H2O, 2.50 mM MgSO4•7H2O, 1.45 mM KH2PO4, while content of micronutrients was 0.233 mM FeNa-EDTA, 0.215 mM MnSO4•4H2O and without CuSO4•5H2O. Increased sucrose concentration to 4.00% (w/v in modified MS liquid medium could increase total of two-alkaloid. The modification of macronutrients and micronutrients concentration based the optimum production of biomass was obtained MSBs medium that producing high biomass but also increasing the production of 9-hydroxycanthin-6-one. The modification of macronutrients or macronutrients and micronutrients based the optimum total of two-alkaloid was obtained MSC and MSD medium that producing low fresh weight but producing the high 9-hydroxycanthin-6-one.

  3. Quantitative Characterization of the Growth of Deinococcus geothermalis DSM-11302: Effect of Inoculum Size, Growth Medium and Culture Conditions.

    Science.gov (United States)

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

    2015-08-20

    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.

  4. 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

    2006-01-01

    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.

  5. Optimization of a Culture Medium for Increased Mevinolin Production by Aspergillus terreus Strain

    Directory of Open Access Journals (Sweden)

    Atalla, M. M.

    2008-01-01

    Full Text Available Fungi are important sources for the production of some pharmaceutical compounds. e.g. lovastatin, mevinolin and monacolin K. These are competitive inhibitors of 3-hydroxy-3-methyl glutaryl coenzyme A reductase, the rate limiting enzyme of cholesterol synthesis pathway. Four fungal strains of Aspergillus terreus and one Penicillium patulum were tested for their potential to produce mevinolin. The fungal strains were cultivated in four different semi-synthetic media to select a fermentation medium and a fungal strain that has the ability to secret mevinolin in high yield. The fermentation followed by TLC screening. Positive results were evaluated by confirmatory HPLC. A. terreus J9 was the best strain for producing mevinolin with a level of 148.66 mg/L of Dox-rice medium. Cultivation a 7.5 L in fermenter, A. terreus J9 produced 932.15 mg/L after 96 h using Dox-rice medium at 6.5 pH. Rise in acidity or alkalinity decrease mevinolin producing ability. Ammonium sulphate in the medium as sulphur and nitrogen sources influenced greatly mevinolin production as well as incubation period. Maximum production was obtained after 36 h incubation. The maximum value of the mevinolin concentratiom (1761.6 mg/L was attained at 400rpm after 60 h fermentation at 28 ºC. The optimized medium resulted in a significant increase of mevinolin cocentration, as compared with that obtained by the fermentation of many other A. terreus species.

  6. The presence of cortical neurons in striatal-cortical co-cultures alters the effects of dopamine and BDNF on Medium Spiny Neuron dendritic development

    Directory of Open Access Journals (Sweden)

    Rachel D Penrod

    2015-07-01

    Full Text Available Medium spiny neurons (MSNs are the major striatal neuron and receive synaptic input from both glutamatergic and dopaminergic afferents. These synapses are made on MSN dendritic spines, which undergo density and morphology changes in association with numerous disease and experience-dependent states. Despite wide interest in the structure and function of mature MSNs, relatively little is known about MSN development. Furthermore, most in vitro studies of MSN development have been done in simple striatal cultures that lack any type of non-autologous synaptic input, leaving open the question of how MSN development is affected by a complex environment that includes other types of neurons, glia, and accompanying secreted and cell-associated cues. Here we characterize the development of MSNs in striatal-cortical co-culture, including quantitative morphological analysis of dendritic arborization and spine development, describing progressive changes in density and morphology of developing spines. Overall, MSN growth is much more robust in the striatal-cortical co-culture compared to striatal mono-culture. Inclusion of dopamine in the co-culture further enhances MSN dendritic arborization and spine density, but the effects of dopamine on dendritic branching are only significant at later times in development. In contrast, exogenous Brain Derived Neurotrophic Factor (BDNF has only a minimal effect on MSN development in the co-culture, but significantly enhances MSN dendritic arborization in striatal mono-culture. Importantly, inhibition of NMDA receptors in the co-culture significantly enhances the effect of exogenous BDNF, suggesting that the efficacy of BDNF depends on the cellular environment. Combined, these studies identify specific periods of MSN development that may be particularly sensitive to perturbation by external factors and demonstrate the importance of studying MSN development in a complex signaling environment.

  7. Suggestions on how to Improve Employee Motivation in Chinese Small and Medium-Sized Private Enterprises. Case Company: X Glassware Factory.

    OpenAIRE

    Li, Yufei

    2016-01-01

    With the economic revolution and open economy policy, China’s private enterprises have become the main driver of the economic growth, covering the main areas of China. However, with the development of enterprises and competition, more and more small and medium-sized private enterprises have faced with serious challenges in human resource management and employee motivation. The phenomena of low employees’ satisfaction,poor motivation and the loss of labor force are prevalent, which have a nega...

  8. Culture conditions for yellow pigment formation byMonascus sp. KB 10 grown on cassava medium.

    Science.gov (United States)

    Yongsmith, B; Tabloka, W; Yongmanitchai, W; Bavavoda, R

    1993-01-01

    An isolate ofMonascus, from a commercial, fermented soybean curd (sufu) was grown on a cassava medium. With medium at an initial pH of 7.0 an orange-red pigmentation was produced but with an initial pH below 4, a light golden pigment was obtained. A medium containing, w/v, 3% cassava starch, 0.4% peptone and 0.1% glutamic acid, with an initial pH of 2.5 was optimal for the production of this yellow pigment, which had a single maximum absorption spectrum at 330 nm. The spectroscopic characterization of the purified yellow pigments demonstrated a monascin-ankaflavin-monascorubrin skeleton.

  9. A simple colony-formation assay in liquid medium, termed 'tadpoling', provides a sensitive measure of Saccharomyces cerevisiae culture viability.

    Science.gov (United States)

    Welch, Aaron Z; Koshland, Douglas E

    2013-12-01

    Here we describe the first high-throughput amenable method of quantifying Saccharomyces cerevisiae culture viability. Current high-throughput methods of assessing yeast cell viability, such as flow cytometry and SGA analysis, do not measure the percentage viability of a culture but instead measure cell vitality or colony fitness, respectively. We developed a method, called tadpoling, to quantify the percentage viability of a yeast culture, with the ability to detect as few as one viable cell amongst ~10(8) dead cells. The most important feature of this assay is the exploitation of yeast colony formation in liquid medium. Utilizing a microtiter dish, we are able to observe a range of viability of 100% to 0.0001%. Comparison of tadpoling to the traditional plating method to measure yeast culture viability reveals that, for the majority of Saccharomyces species analyzed there is no significant difference between the two methods. In comparison to flow cytometry using propidium iodide, the high-throughput method of measuring yeast culture viability, tadpoling is much more accurate at culture viabilities viability.

  10. 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

    2015-12-01

    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.

  11. Optimization of culturing condition and medium composition for the production of alginate lyase by a marine Vibrio sp. YKW-34

    Science.gov (United States)

    Fu, Xiaoting; Lin, Hong; Kim, Sang Moo

    2008-02-01

    Carbohydrases secreted by marine Vibrio sp. YKW-34 with strong Laminaria cell wall degrading ability were screened, and among them alginate lyase was found to be dominant. The effects of medium composition and culturing condition on the production of alginate lyase by marine Vibrio sp. YKW-34 in flask were investigated in this study. In the culture medium of marine broth, no alginate lyase was produced. The activity of the alginate lyase, after being induced, reached 5 UmL-1. The best inoculum volume and inoculum age were 10% and 12 h, respectively. The optimal temperature for alginate lyase production was 25°C. The fermentation medium was composed of 0.5% of Laminaria powder and 0.2% of KNO3 with an initial acidity of pH 8.0. Alginate could induce alginate lyase production but not as efficiently as Laminaria powder did. The addition of fucoidan, cellulose and glucose had negative effect on the alginate lyase production. Other kinds of nitrogen sources, such as yeast extract, beef extract and peptone, had positive effect on the growth of the microorganism and negative effect on alginate lyase production. In addition, the time course of alginate lyase production under the optimized condition was described. The optimal harvest time was 48 h.

  12. Evaluation of dry sheet medium culture plate (Compactdry TC) method for determining numbers of bacteria in food samples.

    Science.gov (United States)

    Mizuochi, S; Kodaka, H

    2000-05-01

    The Compactdry, a ready-to-use and self-diffusible dry medium sheet culture system, has been developed by the Nissui Pharmaceutical Co. Ltd. for enumerating bacteria in food. The Compactdry consists of special spread sheet with culture medium that is the same as standard method nutrients, a cold water-soluble gelling agent, and a unique plastic dish. The procedure for bacterial examination in a sample solution (1 ml) is to just inoculate a test solution into the center of the self-diffusible medium and incubate at 35 degrees C for 48 h. The Compactdry TC (CTC) for the enumeration of total aerobic bacteria from 97 food samples was compared with the standard plate count (SPC) method and 3M Petrifilm aerobic count plates (PAC). The correlation coefficients between the CTC and SPC method, the CTC and PAC, and the PAC and SPC method were 0.97, 0.99, and 0.97, respectively. The Compactdry system is useful for the enumeration of total aerobic bacteria in food and may be a possible suitable alternative to the conventional pour-plate or the Petrifilm plate methods.

  13. Influence of the Culture Medium in Dose-Response Effect of the Chlorhexidine on Streptococcus mutans Biofilms

    Directory of Open Access Journals (Sweden)

    Vanessa Salvadego de Queiroz

    2016-01-01

    Full Text Available The aim of this study was to evaluate the influence of culture medium on dose-response effect of chlorhexidine (CHX on Streptococcus mutans UA159 biofilm and validate the use of the cation-adjusted-Müller-Hinton broth (MH for the evaluation of antibacterial activity. Ultrafiltered Tryptone-Yeast Extract Broth (UTYEB was compared against MH and MH with blood supplementation (MHS. For each medium, six groups (n=4 were assessed: two negative control groups (baseline 48 and 120 h and four experimental groups (0.0001, 0.001, 0.012, and 0.12% CHX. S. mutans biofilm grew on glass slides of each media containing 1% sucrose. After 48 h of growth, biofilms of baseline 48 h were collected and the other groups were treated for 1 min, twice a day, for 3 days, with their respective treatments. The media were changed daily and pH was measured. After 120 h, biofilms were collected and dry weight and viable microorganisms were determined. Results showed CHX dose-response effect being observed in all media for all the variables. However, MH and MHS showed higher sensitivity than UTYEB (p<0.05. We can conclude that the culture medium does influence dose-response effect of CHX on Streptococcus mutans biofilm and that MH can be used for antibacterial activity.

  14. Optimization of Culturing Condition and Medium Composition for the Production of Alginate Lyase by a Marine Vibrio sp. YKW-34

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Carbohydrases secreted by marine Vibrio sp. YKW-34 with strong Laminaria cell wall degrading ability were screened,and among them alginate lyase was found to be dominant. The effects of medium composition and culturing condition on the production of alginate lyase by marine Vibrio sp. YKW-34 in flask were investigated in this study. In the culture medium of marine broth, no alginate lyase was produced. The activity of the alginate lyase, after being induced, reached 5 UmL-1. The best inoculum volume and inoculum age were 10% and 12 h, respectively. The optimal temperature for alginate lyase production was 25℃. The fermentation medium was composed of 0.5% of Laminaria powder and 0.2% of KNO3 with an initial acidity of pH 8.0. Alginate could induce alginate lyase production but not as efficiently as Laminaria powder did. The addition of fucoidan, cellulose and glucose had negative effect on the alginate lyase production. Other kinds of nitrogen sources, such as yeast extract, beef extract and peptone, had positive effect on the growth of the microorganism and negative effect on alginate lyase production. In addition, the time course of alginate lyase production under the optimized condition was described. The optimal harvest time was 48 h.

  15. Influence of the Culture Medium in Dose-Response Effect of the Chlorhexidine on Streptococcus mutans Biofilms

    Science.gov (United States)

    de Queiroz, Vanessa Salvadego; Ccahuana-Vásquez, Renzo Alberto; Tedesco, Alcides Fabiano; Lyra, Luzia; Cury, Jaime Aparecido; Schreiber, Angélica Zaninelli

    2016-01-01

    The aim of this study was to evaluate the influence of culture medium on dose-response effect of chlorhexidine (CHX) on Streptococcus mutans UA159 biofilm and validate the use of the cation-adjusted-Müller-Hinton broth (MH) for the evaluation of antibacterial activity. Ultrafiltered Tryptone-Yeast Extract Broth (UTYEB) was compared against MH and MH with blood supplementation (MHS). For each medium, six groups (n = 4) were assessed: two negative control groups (baseline 48 and 120 h) and four experimental groups (0.0001, 0.001, 0.012, and 0.12% CHX). S. mutans biofilm grew on glass slides of each media containing 1% sucrose. After 48 h of growth, biofilms of baseline 48 h were collected and the other groups were treated for 1 min, twice a day, for 3 days, with their respective treatments. The media were changed daily and pH was measured. After 120 h, biofilms were collected and dry weight and viable microorganisms were determined. Results showed CHX dose-response effect being observed in all media for all the variables. However, MH and MHS showed higher sensitivity than UTYEB (p < 0.05). We can conclude that the culture medium does influence dose-response effect of CHX on Streptococcus mutans biofilm and that MH can be used for antibacterial activity. PMID:27293967

  16. Influence of the Culture Medium in Dose-Response Effect of the Chlorhexidine on Streptococcus mutans Biofilms.

    Science.gov (United States)

    de Queiroz, Vanessa Salvadego; Ccahuana-Vásquez, Renzo Alberto; Tedesco, Alcides Fabiano; Lyra, Luzia; Cury, Jaime Aparecido; Schreiber, Angélica Zaninelli

    2016-01-01

    The aim of this study was to evaluate the influence of culture medium on dose-response effect of chlorhexidine (CHX) on Streptococcus mutans UA159 biofilm and validate the use of the cation-adjusted-Müller-Hinton broth (MH) for the evaluation of antibacterial activity. Ultrafiltered Tryptone-Yeast Extract Broth (UTYEB) was compared against MH and MH with blood supplementation (MHS). For each medium, six groups (n = 4) were assessed: two negative control groups (baseline 48 and 120 h) and four experimental groups (0.0001, 0.001, 0.012, and 0.12% CHX). S. mutans biofilm grew on glass slides of each media containing 1% sucrose. After 48 h of growth, biofilms of baseline 48 h were collected and the other groups were treated for 1 min, twice a day, for 3 days, with their respective treatments. The media were changed daily and pH was measured. After 120 h, biofilms were collected and dry weight and viable microorganisms were determined. Results showed CHX dose-response effect being observed in all media for all the variables. However, MH and MHS showed higher sensitivity than UTYEB (p culture medium does influence dose-response effect of CHX on Streptococcus mutans biofilm and that MH can be used for antibacterial activity.

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

    Science.gov (United States)

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

    2014-03-01

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

  18. Improvement on light penetrability and microalgae biomass production by periodically pre-harvesting Chlorella vulgaris cells with culture medium recycling.

    Science.gov (United States)

    Huang, Yun; Sun, Yahui; Liao, Qiang; Fu, Qian; Xia, Ao; Zhu, Xun

    2016-09-01

    To improve light penetrability and biomass production in batch cultivation, a cultivation mode that periodically pre-harvesting partial microalgae cells from suspension with culture medium recycling was proposed. By daily pre-harvesting 30% microalgae cells from the suspension, the average light intensity in the photobioreactor (PBR) was enhanced by 27.05-122.06%, resulting in a 46.48% increase in total biomass production than that cultivated in batch cultivation without pre-harvesting under an incident light intensity of 160μmolm(-2)s(-1). Compared with the semi-continuous cultivation with 30% microalgae suspension daily replaced with equivalent volume of fresh medium, nutrients and water input was reduced by 60% in the proposed cultivation mode but with slightly decrease (12.82%) in biomass production. No additional nutrient was replenished when culture medium recycling. Furthermore, higher pre-harvesting ratios (40%, 60%) and lower pre-harvesting frequencies (every 2, 2.5days) were not advantageous for the pre-harvesting cultivation mode. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Innovative culture, management control systems and performance in small and medium-sized Spanish family firms

    Directory of Open Access Journals (Sweden)

    Antonio Duréndez

    2011-06-01

    Full Text Available The aim of this paper is to research on organizational culture and management of family firms. We identify family-firms innovative culture and assess the relationship between organizational culture, management control systems (MCS use and their effects on performance of SME family-firms. With this purpose, we carry out an empirical analysis on a sample of Spanish SMEs (285 family and 151 non-family firms. Results show that (1 family-firms have a more hierarchical culture and a lesser extent of MCS use than non-family firms have, and (2 an innovative culture and the use of MCS have positive influences on family-firm performance.

  20. Teaching Sociolinguistics: A Medium for Cultural Awareness of Indonesian University Foreign Language Learners

    Directory of Open Access Journals (Sweden)

    Agnes Herawati

    2014-04-01

    Full Text Available This paper tries to show the evidences that indicate how teaching Sociolinguistics can result in a number of valuable outcomes, including helping students understand and appreciate other cultures different from theirs. Sociolinguistics provides useful examples of language usage in different genres, including how culture influences people in using a language. The opportunities of learning other cultures through language will take the students to the higher level of appreciation of the culture of the target language. To determine how this outcome can be achieved in the language classrooms, this paper provides a review of closely connected literature about how to bridge the gap between cultures in particular. However, to increase its completeness and relevance, this paper also provides some research results that reveal how teaching Sociolinguistics has taken its new applicability and importance, and furthermore adds the effects on how students become more proficient and enthusiastic about their learning. 

  1. Tissue culture of human alveolar periosteal sheets using a stem-cell culture medium (MesenPRO-RS™): In vitro expansion of CD146-positive cells and concomitant upregulation of osteogenic potential in vivo.

    Science.gov (United States)

    Uematsu, Kohya; Kawase, Tomoyuki; Nagata, Masaki; Suzuki, Kenji; Okuda, Kazuhiro; Yoshie, Hiromasa; Burns, Douglas M; Takagi, Ritsuo

    2013-01-01

    We have previously demonstrated that multilayered periosteal sheets prepared from the explant culture of alveolar periosteum serve as a promising osteogenic grafting material in periodontal tissue regeneration. For the preparation of more potent periosteal sheets, we examined the applicability of stem-cell culture media. Compared to the control medium (Medium 199+10% FBS), periosteal sheets expanded with MesenPRO-RS™ medium exhibited these features: Cells grew three-dimensionally and deposited collagen in the extracellular spaces to form thicker multilayers of cells. Chondrocytic markers were not significantly upregulated. Contractile force was generated in proportion with the increased thickness of the periosteal sheets and the formation of cytoplasmic α-smooth muscle actin fibers. However, myofibroblastic markers were not significantly upregulated. The surface marker CD146 was substantially upregulated, while both CD73 and CD105 were downregulated. Alkaline phosphatase, a representative osteoblastic marker, was not upregulated by osteogenic induction. However, these expanded periosteal sheets exhibited substantially stronger osteogenic differentiation when implanted in nude mice. Therefore, despite our reservations, MesenPRO medium effectively expanded the cells contained in periosteal sheets to promote the formation of thicker multilayers of cells in vitro, and these enhanced periosteal sheets expressed increased osteogenic potential at implantation sites in vivo. In conjunction with data indicating that CD146-positive cells were notably expanded and the recently proposed concept that CD146 is a marker for osteogenic progenitor cells found in the bone marrow stroma, our findings suggest that MesenPRO medium improves the preparation of highly osteogenic periosteal sheets suitable for clinical application largely through the induction of CD146-positive cells. Copyright © 2012 Elsevier B.V. All rights reserved.

  2. Evaluation of Urea-motility-indole medium for recognition and differentiation of Salmonella and Shigella species in stool cultures.

    Science.gov (United States)

    Rosa Fraile, M; Vega Aleman, D; Fernandez Gutierrez, C

    1980-09-01

    A semisolid urea-motility-indole medium designed for detection in Enterobacteriaceae of urease activity, motility, and indole production in one tube was prepared and evaluated. The formulation of the medium was similar to that of Christensen urea agar, but the agar concentration was 0.2%, and 1% tryptone was added. Results with 687 strains of Enterobacteriaceae were the same as those obtained with standard test media (98% overall agreement). The urea-motility-indole medium was also used in combination with Kligler iron agar for the recognition and differentiation of Salmonella and Shigella species from colonies picked from plating media in fecal cultures. This combination was compared with the combination of Kligler iron agar and lysine iron agar with 507 strains of non-lactose-fermenting Enterobacteriaceae. Although both combinations enabled the presumptive recognition and differentiation of Salmonella and Shigella species, an analysis of data indicated that the combination of Kligler iron agar and urea-motility-indole medium performed better than the combination of Kligler iron agar and lysine iron agar in detecting Salmonella and Shigella species.

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

    Science.gov (United States)

    Idelevich, E A; Grünastel, B; Peters, G; Becker, K

    2016-03-01

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

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

    DEFF Research Database (Denmark)

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

    2005-01-01

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

  5. [The optimization of the nitric oxide quantitative analysis for its determination in the cultural medium of mammalian cell culture].

    Science.gov (United States)

    Akimov, M G; Fomina-Ageeva, E V; Bezuglov, V V

    2015-01-01

    The protocol for the quantitative analysis of nitric oxide as nitrite-ion suitable for determination of its production by a mammalian cell culture was developed. The optimal results were obtained using microvolume-adjusted Griess method after the preliminary reduction of NO3- to NO2- with non-activated cadmium. The protocol was verified on a rat glioma C6 cell culture. The developed method may be used for the nitric oxide determination in 96-well and 48-well microplates; the detection limit is 2.1 ± 0.1 μM for NO2- and 2.9 ± 0.1 μM for NO3-.

  6. Degradation of Bunker C Fuel Oil by White-Rot Fungi in Sawdust Cultures Suggests Potential Applications in Bioremediation.

    Directory of Open Access Journals (Sweden)

    Darcy Young

    Full Text Available Fungal lignocellulolytic enzymes are promising agents for oxidizing pollutants. This study investigated degradation of Number 6 "Bunker C" fuel oil compounds by the white-rot fungi Irpex lacteus, Trichaptum biforme, Phlebia radiata, Trametes versicolor, and Pleurotus ostreatus (Basidiomycota, Agaricomycetes. Averaging across all studied species, 98.1%, 48.6%, and 76.4% of the initial Bunker C C10 alkane, C14 alkane, and phenanthrene, respectively were degraded after 180 days of fungal growth on pine media. This study also investigated whether Bunker C oil induces changes in gene expression in the white-rot fungus Punctularia strigosozonata, for which a complete reference genome is available. After 20 days of growth, a monokaryon P. strigosozonata strain degraded 99% of the initial C10 alkane in both pine and aspen media but did not affect the amounts of the C14 alkane or phenanthrene. Differential gene expression analysis identified 119 genes with ≥ log2(2-fold greater expression in one or more treatment comparisons. Six genes were significantly upregulated in media containing oil; these genes included three enzymes with potential roles in xenobiotic biotransformation. Carbohydrate metabolism genes showing differential expression significantly accumulated transcripts on aspen vs. pine substrates, perhaps reflecting white-rot adaptations to growth on hardwood substrates. The mechanisms by which P. strigosozonata may degrade complex oil compounds remain obscure, but degradation results of the 180-day cultures suggest that diverse white-rot fungi have promise for bioremediation of petroleum fuels.

  7. Degradation of Bunker C Fuel Oil by White-Rot Fungi in Sawdust Cultures Suggests Potential Applications in Bioremediation.

    Science.gov (United States)

    Young, Darcy; Rice, James; Martin, Rachael; Lindquist, Erika; Lipzen, Anna; Grigoriev, Igor; Hibbett, David

    2015-01-01

    Fungal lignocellulolytic enzymes are promising agents for oxidizing pollutants. This study investigated degradation of Number 6 "Bunker C" fuel oil compounds by the white-rot fungi Irpex lacteus, Trichaptum biforme, Phlebia radiata, Trametes versicolor, and Pleurotus ostreatus (Basidiomycota, Agaricomycetes). Averaging across all studied species, 98.1%, 48.6%, and 76.4% of the initial Bunker C C10 alkane, C14 alkane, and phenanthrene, respectively were degraded after 180 days of fungal growth on pine media. This study also investigated whether Bunker C oil induces changes in gene expression in the white-rot fungus Punctularia strigosozonata, for which a complete reference genome is available. After 20 days of growth, a monokaryon P. strigosozonata strain degraded 99% of the initial C10 alkane in both pine and aspen media but did not affect the amounts of the C14 alkane or phenanthrene. Differential gene expression analysis identified 119 genes with ≥ log2(2-fold) greater expression in one or more treatment comparisons. Six genes were significantly upregulated in media containing oil; these genes included three enzymes with potential roles in xenobiotic biotransformation. Carbohydrate metabolism genes showing differential expression significantly accumulated transcripts on aspen vs. pine substrates, perhaps reflecting white-rot adaptations to growth on hardwood substrates. The mechanisms by which P. strigosozonata may degrade complex oil compounds remain obscure, but degradation results of the 180-day cultures suggest that diverse white-rot fungi have promise for bioremediation of petroleum fuels.

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

    Directory of Open Access Journals (Sweden)

    Fábio de Farias Neves

    2013-07-01

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

  9. Influence of Selenium Content in the Culture Medium on Protein Profile of Yeast Cells Candida utilis ATCC 9950

    Directory of Open Access Journals (Sweden)

    Marek Kieliszek

    2015-01-01

    Full Text Available Selenium is an essential trace element for human health and it has been recognized as a component of several selenoproteins with crucial biological functions. It has been identified as a component of active centers of many enzymes, as well as integral part of biologically active complexes. The aim of the study was to evaluate the protein content and amino acid profile of the protein of fodder yeast Candida utilis ATCC 9950 cultured in media control and experimental enriched selenium. Protein analysis was performed using SDS-PAGE method consisting of polyacrylamide gel electrophoresis in the presence of SDS. The highest contents of soluble protein (49,5 mg/g were found in yeast cells after 24-hour culture conducted in control (YPD medium. In the presence of selenium there were determined small amounts of protein content. With increasing time of yeast culture (to 72 hours the control and experimental media were reported to reduce soluble protein content. In electropherogram proteins from control cultures was observed the presence of 10 protein fractions, but in all the experimental cultures (containing 20, 30, and 40 mg/L selenium of 14 protein fractions. On the basis of the molecular weights of proteins, it can be concluded that they were among others: selenoprotein 15 kDa and selenoprotein 18 kDa.

  10. Teaching Sociolinguistics: A Medium for Cultural Awareness of Indonesian University Foreign Language Learners

    National Research Council Canada - National Science Library

    Agnes Herawati

    2014-01-01

    This paper tries to show the evidences that indicate how teaching Sociolinguistics can result in a number of valuable outcomes, including helping students understand and appreciate other cultures different from theirs...

  11. Information security culture: a survey in small and medium sized enterprises

    OpenAIRE

    Lopes, Isabel Maria; P. De Oliveira

    2014-01-01

    Information security is a relevant fact for current organizations. There are factors inextricably linked to this issue, and one cannot talk about information security in an organization without addressing and understanding the information security culture of that institution. Maximizing the organizational culture within an organization will enable the safeguard of information security. For that, we need to understand which the inhibiting and the enabling factors are...

  12. Understanding information security culture: a survey in small and medium sized enterprises

    OpenAIRE

    Lopes, Isabel Maria; P. De Oliveira

    2014-01-01

    Information security is a relevant fact for current organizations. There are factors inextricably linked to this issue, and one cannot talk about information security in an organization without addressing and understanding the information security culture of that institution. Maximizing the organizational culture within an organization will enable the safeguard of information security. For that, we need to understand which the inhibiting and the enabling factors are. This paper contributes to...

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

    Directory of Open Access Journals (Sweden)

    A J Kindo

    2016-01-01

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

  14. Optimization of a serum-free culture medium for mouse embryonic stem cells using design of experiments (DoE) methodology

    OpenAIRE

    Knöspel, Fanny; Schindler, Rudolf K.; Lübberstedt, Marc; Petzolt, Stephanie; Gerlach, Jörg C.; Zeilinger, Katrin

    2010-01-01

    The in vitro culture behaviour of embryonic stem cells (ESC) is strongly influenced by the culture conditions. Current culture media for expansion of ESC contain some undefined substances. Considering potential clinical translation work with such cells, the use of defined media is desirable. We have used Design of Experiments (DoE) methods to investigate the composition of a serum-free chemically defined culture medium for expansion of mouse embryonic stem cells (mESC). Factor screening analy...

  15. THE INFLUENCE OF E-MANAGEMENT APPLICATION UPOIN THE SMALL AND MEDIUM ENTERPRISES ORGANIZATIONAL CULTURE

    OpenAIRE

    Corneliu Gârjoabă

    2011-01-01

    The internet lays the foundation of the knowledge society’s manifestation bringing new opportunities for the development of the business environment, capitalized predominantly by the small and medium enterprises that, benefiting of a flexible structure, cross with great ease to the ,,e-business,, type of business in order to operate on the virtual market and to obtain higher revenues and faster than in the traditional way.

  16. Screening and optimization of low-cost medium for Pseudomonas putida Rs-198 culture using RSM.

    Science.gov (United States)

    Peng, Yanjie; He, Yanhui; Wu, Zhansheng; Lu, Jianjiang; Li, Chun

    2014-01-01

    The plant growth-promoting rhizobacterial strain Pseudomonas putida Rs-198 was isolated from salinized soils from Xinjiang Province. We optimized the composition of the low-cost medium of P. putida Rs-198 based on its bacterial concentration, as well as its phosphate-dissolving and indole acetic acid (IAA)-producing capabilities using the response surface methodology (RSM), and a mathematical model was developed to show the effect of each medium component and its interactions on phosphate dissolution and IAA production. The model predicted a maximum phosphate concentration in medium containing 63.23 mg/L inorganic phosphate with 49.22 g/L corn flour, 14.63 g/L soybean meal, 2.03 g/L K₂HPO₄, 0.19 g/L MnSO₄ and 5.00 g/L NaCl. The maximum IAA concentration (18.73 mg/L) was predicted in medium containing 52.41 g/L corn flour, 15.82 g/L soybean meal, 2.40 g/L K₂HPO₄, 0.17 g/L MnSO₄ and 5.00 g/L NaCl. These predicted values were also verified through experiments, with a cell density of 10(13) cfu/mL, phosphate dissolution of 64.33 mg/L, and IAA concentration of 18.08 mg/L. The excellent correlation between predicted and measured values of each model justifies the validity of both the response models. The study aims to provide a basis for industrialized fermentation using P. putida Rs-198.

  17. Screening and optimization of low-cost medium for Pseudomonas putida Rs-198 culture using RSM

    Directory of Open Access Journals (Sweden)

    Yanjie Peng

    2014-12-01

    Full Text Available The plant growth-promoting rhizobacterial strain Pseudomonas putida Rs-198 was isolated from salinized soils from Xinjiang Province. We optimized the composition of the low-cost medium of P. putida Rs-198 based on its bacterial concentration, as well as its phosphate-dissolving and indole acetic acid (IAA-producing capabilities using the response surface methodology (RSM, and a mathematical model was developed to show the effect of each medium component and its interactions on phosphate dissolution and IAA production. The model predicted a maximum phosphate concentration in medium containing 63.23 mg/L inorganic phosphate with 49.22 g/L corn flour, 14.63 g/L soybean meal, 2.03 g/L K2HPO4, 0.19 g/L MnSO4 and 5.00 g/L NaCl. The maximum IAA concentration (18.73 mg/L was predicted in medium containing 52.41 g/L corn flour, 15.82 g/L soybean meal, 2.40 g/L K2HPO4, 0.17 g/L MnSO4 and 5.00 g/L NaCl. These predicted values were also verified through experiments, with a cell density of 10(13 cfu/mL, phosphate dissolution of 64.33 mg/L, and IAA concentration of 18.08 mg/L. The excellent correlation between predicted and measured values of each model justifies the validity of both the response models. The study aims to provide a basis for industrialized fermentation using P. putida Rs-198.

  18. Butanol and hexanol production in Clostridium carboxidivorans syngas fermentation: Medium development and culture techniques.

    Science.gov (United States)

    Phillips, John R; Atiyeh, Hasan K; Tanner, Ralph S; Torres, Juan R; Saxena, Jyotisna; Wilkins, Mark R; Huhnke, Raymond L

    2015-08-01

    Clostridium carboxidivorans was grown on model syngas (CO:H2:CO2 [70:20:10]) in a defined nutrient medium with concentrations of nitrogen, phosphate and trace metals formulated to enhance production of higher alcohols. C. carboxidivorans was successfully grown in a limited defined medium (no yeast extract, no MES buffer and minimal complex chemical inputs) using an improved fermentation protocol. Low partial pressure of CO in the headspace, coupled with restricted mass transfer for CO and H2, was required for successful fermentation. In the absence of substrate inhibition (particularly from CO), growth limitation increased production of alcohols, especially butanol and hexanol. Concentrations of butanol (over 1.0g/L), hexanol (up to 1.0g/L) and ethanol (over 3.0g/L) were achieved in bottle fermentations. Minimal medium and controlled supply of CO and H2 should be used in characterizing candidate butanol and hexanol producing strains to select for commercial potential.

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

    Directory of Open Access Journals (Sweden)

    Markov S.L.

    2012-01-01

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

  20. Inhibitory synapse formation in a co-culture model incorporating GABAergic medium spiny neurons and HEK293 cells stably expressing GABAA receptors.

    Science.gov (United States)

    Brown, Laura E; Fuchs, Celine; Nicholson, Martin W; Stephenson, F Anne; Thomson, Alex M; Jovanovic, Jasmina N

    2014-11-14

    Inhibitory neurons act in the central nervous system to regulate the dynamics and spatio-temporal co-ordination of neuronal networks. GABA (γ-aminobutyric acid) is the predominant inhibitory neurotransmitter in the brain. It is released from the presynaptic terminals of inhibitory neurons within highly specialized intercellular junctions known as synapses, where it binds to GABAA receptors (GABAARs) present at the plasma membrane of the synapse-receiving, postsynaptic neurons. Activation of these GABA-gated ion channels leads to influx of chloride resulting in postsynaptic potential changes that decrease the probability that these neurons will generate action potentials. During development, diverse types of inhibitory neurons with distinct morphological, electrophysiological and neurochemical characteristics have the ability to recognize their target neurons and form synapses which incorporate specific GABAARs subtypes. This principle of selective innervation of neuronal targets raises the question as to how the appropriate synaptic partners identify each other. To elucidate the underlying molecular mechanisms, a novel in vitro co-culture model system was established, in which medium spiny GABAergic neurons, a highly homogenous population of neurons isolated from the embryonic striatum, were cultured with stably transfected HEK293 cell lines that express different GABAAR subtypes. Synapses form rapidly, efficiently and selectively in this system, and are easily accessible for quantification. Our results indicate that various GABAAR subtypes differ in their ability to promote synapse formation, suggesting that this reduced in vitro model system can be used to reproduce, at least in part, the in vivo conditions required for the recognition of the appropriate synaptic partners and formation of specific synapses. Here the protocols for culturing the medium spiny neurons and generating HEK293 cells lines expressing GABAARs are first described, followed by detailed

  1. Optimization of culture medium composition for manganese peroxidase and tyrosinase production during Reactive Black 5 decolourization by the yeast Trichosporon akiyoshidainum.

    Science.gov (United States)

    Martorell, María M; Pajot, Hipólito F; Rovati, José I; Figueroa, Lucía I C

    2012-03-01

    Decolourization and degradation of the diazo dye Reactive Black 5 was carried out by the yeast Trichosporon akiyoshidainum. A nine-factor Plackett-Burman design was employed for the study and optimization of the decolourization process and production of manganese peroxidase (MnP) and tyrosinase activities. In the present study, 26 individual experiments were conducted and three responses were evaluated. Raising yeast extract concentration significantly enhanced decolourization and MnP production. Carbon and nitrogen sources, glucose and (NH4)2 SO4, showed no significant effect on any response over the concentration range tested. Other culture medium components, such as CaCl2 or MgSO4, could be excluded from the medium formula, as they had no effect on the evaluated responses. Metal ions (Fe, Cu and Mn) showed different effects on decolourization and enzymatic activities. Addition of copper significantly enhanced MnP activity and decreased dye decolourization. On the contrary, iron had a positive effect on decolourization and no effect on enzyme production. Oddly, increasing manganese concentration had a positive effect on tyrosinase production without affecting decolourization or MnP activity. These results strongly suggest that dye decolourization should be regarded as a complex multi-enzymatic process, where optimal medium composition should arise as a compromise between those optimal for each implied enzyme production. Copyright © 2012 John Wiley & Sons, Ltd.

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

    Science.gov (United States)

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

    2012-01-01

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

  3. Long-term, hormone-responsive organoid cultures of human endometrium in a chemically defined medium.

    Science.gov (United States)

    Turco, Margherita Y; Gardner, Lucy; Hughes, Jasmine; Cindrova-Davies, Tereza; Gomez, Maria J; Farrell, Lydia; Hollinshead, Michael; Marsh, Steven G E; Brosens, Jan J; Critchley, Hilary O; Simons, Benjamin D; Hemberger, Myriam; Koo, Bon-Kyoung; Moffett, Ashley; Burton, Graham J

    2017-05-01

    In humans, the endometrium, the uterine mucosal lining, undergoes dynamic changes throughout the menstrual cycle and pregnancy. Despite the importance of the endometrium as the site of implantation and nutritional support for the conceptus, there are no long-term culture systems that recapitulate endometrial function in vitro. We adapted conditions used to establish human adult stem-cell-derived organoid cultures to generate three-dimensional cultures of normal and decidualized human endometrium. These organoids expand long-term, are genetically stable and differentiate following treatment with reproductive hormones. Single cells from both endometrium and decidua can generate a fully functional organoid. Transcript analysis confirmed great similarity between organoids and the primary tissue of origin. On exposure to pregnancy signals, endometrial organoids develop characteristics of early pregnancy. We also derived organoids from malignant endometrium, and so provide a foundation to study common diseases, such as endometriosis and endometrial cancer, as well as the physiology of early gestation.

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

    Directory of Open Access Journals (Sweden)

    Saxena Vandana

    2017-01-01

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

  5. Culture medium for selective isolation and enumeration of Gram-negative bacteria from ground meats.

    OpenAIRE

    Cyzeska, F J; Seiter, J A; Marks, S N; Jay, J M

    1981-01-01

    We developed a new medium, designated peptone bile amphotericin cycloheximide (PBAC) agar, which contains (per liter) 10 g of peptone, 300 mg of bile salts, 1 mg of amphotericin B, 1 g of cycloheximide, and 15 g of agar. When 21 samples of fresh ground beef were studied and plate count agar counts were used as references, we obtained a mean recovery of 28% of total counts with violet red bile agar overlay, whereas we obtained 48% recovery with PBAC agar. With 12 samples of frozen ground beef,...

  6. Praline metabolism by germinating Lilium longiflorum pollen. I. Labelling of cytoplasmic, wall and culture medium molecules

    Directory of Open Access Journals (Sweden)

    W. V. Dashek

    2014-01-01

    Full Text Available Radioactivity occurs in trithloroacetic acid (TCA-soluble and precipitable, cytoplasm and salt-washed walls following germination of Lilium longiflorum, cv. 'Ace' pollen in medium containing [14C]-proline (Pro. Sephadex gel filtration on G-25 through G-100 was employed to determine whether radioactivity in cytoplasm, wall and growth medium from pollen fed [14C]-Pro or [3H]=Pro plus [14C]-arafbinose (Ara was contained within molecules possessing molecular weights of 5,000 to 100,000 daltones or greater. G-25 elution profiles of a crude cytoplasmic fraction (15,000 X g supernatant from [14C]-Pro labelled pollen yielded a radioctive void volume peak and a retarded peak. The void volume peak contained hydroxyproline (Hyp, and exhibited a coincidence of [3H]-Pro and [14C] -Ara labelling when pollen was double labelled with the two isotopes. This peak also contained radioactivity when pollen was germinated in 2-[3H]-myo-inositol. Germination in medium supplemented with 100 µM 2,2'-dipyridyl eliminated radioactivity from 2-[3H]-myo-inositol or [14C]-,Pro in the peak. Filtratian on G-25 of aTCA-soluble fraction of a salt-extract of walls from [14C]-Pro labelled pollen resulted in void volume and two retarded peaks. Void volume and two retarded peaks were also obtained upon G-25 filtration of a cellulase-digest of walls from [M]-Pro labeled pollen. The void volume peak contained Hyp, Lys, Gly, Ala, Ser, Glu and Asp acids, Val, Tyr, Leu or lieu and Pro. Sephadex G-90, 75, and 100 elution profiles of cellulasedigests of walls from [3H]-,Pro and [14C]-Ara labelled pollen yielded radioactive retarded and Hyp-containing void volume peaks with a coincidence of [3H] and [14C] labelling. Label in the void volume was obtained when either rhozyme P11- or pepsin-digests of walls from [14C]-Pro labelled pollen were gel filtered on G-50. Paper electrophoresis coupled with paper chromatography of acid hydrolyzates of salt-washed wall fractions demonstrated 15 of the

  7. A defined mix of cytokines mimics conditioned medium from cultures of bone marrow-derived mesenchymal stem cells and elicits bone regeneration.

    Science.gov (United States)

    Katagiri, Wataru; Sakaguchi, Kohei; Kawai, Takamasa; Wakayama, Yukiko; Osugi, Masashi; Hibi, Hideharu

    2017-06-01

    We previously reported that conditioned medium from cultures of bone marrow-derived mesenchymal stem cells have strong potential to accelerate bone regeneration. We now examine in vitro and in vivo a defined cytokine cocktail that mimics the effects of conditioned medium on bone regeneration. A cocktail of recombinant human insulin-like growth factor-1, vascular endothelial growth factor-A and transforming growth factor-β1 was prepared at concentrations similar to those in conditioned medium. Conversely, these cytokines were depleted from conditioned medium, and the effects of the cocktail, the conditioned medium and the cytokine-depleted conditioned medium on bone regeneration were evaluated in vitro and in vivo. The cytokine cocktail and conditioned medium enhanced cell migration, tube formation, and expression of osteogenic and angiogenic genes. Depletion of cytokines significantly decreased the effects of conditioned medium in vitro. Similarly, the cytokine cocktail and conditioned medium, but not cytokine-depleted medium, increased bone regeneration in damaged rat calvarial bone. Immunohistochemistry indicated that the cytokine cocktail and conditioned medium strongly enhanced recruitment of endogenous stem cells and endothelial cells. The data indicate that the cytokine cocktail and conditioned medium enhance the migration of stem cells and endothelial cells to damaged bone, and elicit osteogenesis and angiogenesis. © 2017 John Wiley & Sons Ltd.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2000-12-07

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

  9. Literature as a Medium of Cultural Memory in Novel by Drago Jančar That Night I Saw Her

    Directory of Open Access Journals (Sweden)

    Primož Tanko

    2015-09-01

    Full Text Available This paper tries to show the literature as an important part of building and conservation of the cultural memory of the society. Cultural memory belongs also to individuals and groups and they have a common reference point, which are often photos or objects that evoke the memory, event or commemoration which is an important part of identity. In this case is also important that literature through biographical or fictional stories represents space and time. Often fictional literature creates a genuine feeling of a memory to some particular times or events, because it had a wider range of stories, characters and situations and in this way is literature creating collective memory in a more concentrated form. On the case of recent novel by Drago Jancar To noč sem jo videl - That night I saw her, will I show how work literature as a medium of cultural memory and also which are the themes, that are important in Slovenian cultural memory

  10. Direct fermentation of xylan by Clostridium strain BOH3 for the production of butanol and hydrogen using optimized culture medium.

    Science.gov (United States)

    Rajagopalan, Gobinath; He, Jianzhong; Yang, Kun Lin

    2014-02-01

    Clostridium strain BOH3 is able to utilize 10 g/l of xylan in reinforced clostridial medium (RCM) and produce butanol and hydrogen. However, increasing xylan concentration to 30 g/l does not enhance the production of butanol and hydrogen due to insufficient expression of xylanase enzyme (27.5 U/mg). To enhance the xylanase activity, an optimized culture medium (OCM), which consists of sugarcane bagasse hydrolysate (11.75 g/l), ammonium sulfate (8.92 g/l) and iron (III) chloride (1.45 mM) is designed. In the optimized OCM, Clostridium strain BOH3 expresses more xylanase and shows higher xylanase activity (44.05 ± 0.25 U/mg) in the OCM. This activity is about 1.6-fold higher than that in the original RCM. Employing OCM as a medium, Clostridium strain BOH3 effectively ferments high concentration (30 and 50 g/l) of xylan and produces 12.05 ± 0.15 and 14.80 ± 0.15 g/l of butanol and 1.78 ± 0.08 and 2.65 ± 0.15 l/l of hydrogen, respectively. Copyright © 2013 Elsevier Ltd. All rights reserved.

  11. Population structure of mixed Mycobacterium tuberculosis infection is strain genotype and culture medium dependent.

    NARCIS (Netherlands)

    Hanekom, M.; Streicher, E.M.; Berg, D. Van den; Cox, H.; McDermid, C.; Bosman, M.; Pittius, N.C. Gey van; Victor, T.C.; Kidd, M.; Soolingen, D. van; Helden, P.D. van; Warren, R.M.

    2013-01-01

    BACKGROUND: Molecular genotyping methods have shown infection with more than one Mycobacterium tuberculosis strain genotype in a single sputum culture, indicating mixed infection. AIM: This study aimed to develop a PCR-based genotyping tool to determine the population structure of M. tuberculosis st

  12. Effects of melatonin on in vitro development of mouse two-cell embryos cultured in HTF medium.

    Science.gov (United States)

    Tian, Xiu-Zhi; Wen, Qing; Shi, Jian-Min; Liang-Wang; Zeng, Shen-Ming; Tian, Jian-Hui; Zhou, Guang-Bin; Zhu, Shi-En; Liu, Guo-Shi

    2010-01-01

    Melatonin is capable of improving the developmental capacity of ovine, porcine and bovine embryos in vitro. However, whether melatonin possesses similar benefits to the in vitro mouse embryonic development has yet to be determined. In this study, we assessed the effects of various concentrations of melatonin (10-13 to 10-3 M) on the in-vitro development of mouse embryos cultured in HTF medium for 96 hr; embryos cultured without melatonin were used as control. The in vitro development of mouse two-cell embryos significantly benefited from treatment with melatonin in a concentration-dependent manner. The effects of melatonin on the rates of blastocyst formation, hatching/hatched blastocysts and cell number per blastocyst were bi-phasic; all significantly increased by melatonin at 10-13 to 10-5 M and decreased by melatonin at 10-3 M. Maximal benefit of melatonin on in vitro mouse 2-cell embryo development was achieved at a concentration of 10-9 M. In comparison to control, 10-9 M melatonin increased blastocyst formation rate from 48.08 +/- 5.25% to 82.08 +/- 2.34% (p HTF medium.

  13. Optimization of Culture Conditions and Medium Composition for the Marine Algicidal Bacterium Alteromonas sp.DH46 by Uniform Design

    Institute of Scientific and Technical Information of China (English)

    LIN Jing; ZHENG Wei; TIAN Yun; WANG Guizhong; ZHENG Tianling

    2013-01-01

    Harmful algal blooms (HABs) have led to extensive ecological and environmental issues and huge economic losses.Various HAB control techniques have been developed,and biological methods have been paid more attention.Algicidal bacteria is a general designation for bacteria which inhibit algal growth in a direct or indirect manner,and kill or damage the algal cells.A metabolite which is strongly toxic to the dinoflagellate Alexandrium tamarense was produced by strain DH46 of the alga-lysing bacterium Alteromonas sp.The culture conditions were optimized using a single-factor test method.Factors including carbon source,nitrogen source,temperature,initial pH value,rotational speed and salinity were studied.The results showed that the cultivation of the bacteria at 28℃ and 180r min-1 with initial pH 7 and 30 salt contcentration favored both the cell growth and the lysing effect of strain DH46.The optimal medium composition for strain DH46 was determined by means of uniform design experimentation,and the most important components influencing the cell density were tryptone,yeast extract,soluble starch,NaNO3 and MgSO4.When the following culture medium was used (tryptone 14.0g,yeast extract 1.63g,soluble starch 5.0g,NaNO3 1.6g,MgSO4 2.3 g in 1L),the largest bacterial dry weight (7.36gL-1) was obtained,which was an enhancement of 107% compared to the initial medium; and the algal lysis rate was as high as 98.4% which increased nearly 10% after optimization.

  14. Optimization of culture conditions and medium composition for the marine algicidal bacterium Alteromonas sp. DH46 by uniform design

    Science.gov (United States)

    Lin, Jing; Zheng, Wei; Tian, Yun; Wang, Guizhong; Zheng, Tianling

    2013-09-01

    Harmful algal blooms (HABs) have led to extensive ecological and environmental issues and huge economic losses. Various HAB control techniques have been developed, and biological methods have been paid more attention. Algicidal bacteria is a general designation for bacteria which inhibit algal growth in a direct or indirect manner, and kill or damage the algal cells. A metabolite which is strongly toxic to the dinoflagellate Alexandrium tamarense was produced by strain DH46 of the alga-lysing bacterium Alteromonas sp. The culture conditions were optimized using a single-factor test method. Factors including carbon source, nitrogen source, temperature, initial pH value, rotational speed and salinity were studied. The results showed that the cultivation of the bacteria at 28°C and 180 r min-1 with initial pH 7 and 30 salt contcentration favored both the cell growth and the lysing effect of strain DH46. The optimal medium composition for strain DH46 was determined by means of uniform design experimentation, and the most important components influencing the cell density were tryptone, yeast extract, soluble starch, NaNO3 and MgSO4. When the following culture medium was used (tryptone 14.0g, yeast extract 1.63g, soluble starch 5.0 g, NaNO3 1.6 g, MgSO4 2.3 g in 1L), the largest bacterial dry weight (7.36 g L-1) was obtained, which was an enhancement of 107% compared to the initial medium; and the algal lysis rate was as high as 98.4% which increased nearly 10% after optimization.

  15. Analysis of CHO cells metabolic redistribution in a glutamate-based defined medium in continuous culture.

    Science.gov (United States)

    Altamirano, C; Illanes, A; Casablancas, A; Gámez, X; Cairó, J J; Gòdia, C

    2001-01-01

    The effect of glutamine replacement by glutamate and the balance between glutamate and glucose metabolism on the redistribution of t-PA-producing recombinant CHO cells metabolism is studied in a series of glucose shift down and shift up experiments in continuous culture. These experiments reveal the existence of multiple steady states, and experimental data are used to perform metabolic flux analysis to gain a better insight into cellular metabolism and its redistribution. Regulation of glucose feed rate promotes a higher efficiency of glucose and nitrogen source utilization, with lower production of metabolic byproducts, but this reduces t-PA specific production rate. This reduction under glucose limitation can be attributed to the fact that the cells are forced to efficiently utilize the carbon and energy source for growth, impairing the production of dispensable metabolites. It is, therefore, the combination of growth rate and carbon and energy source availability that determines the level of t-PA production in continuous culture.

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

    OpenAIRE

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

    2012-01-01

    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...

  17. Effect of culture medium on biocalcification by Pseudomonas Putida, Lysinibacillus Sphaericus and Bacillus Subtilis

    OpenAIRE

    Márcia Aiko Shirakawa; Maria Alba Cincotto; Daniel Atencio; Gaylarde,Christine C.; John,Vanderley M.

    2011-01-01

    The objective of this study is to investigate the efficiency of calcium carbonate bioprecipitation by Lysinibacillus sphaericus, Bacillus subtilis and Pseudomonas putida, obtained from the Coleção de Culturas do Instituto Nacional de Controle de Qualidade em Saúde (INCQS), as a first step in determining their potential to protect building materials against water uptake. Two culture media were studied: modified B4 containing calcium acetate and 295 with calcium chloride. Calcium consumption in...

  18. Comparing two protocols of DNA extraction of Trypanosoma cruzi cultured in axenic medium

    OpenAIRE

    López, Mariela; Instituto de Investigaciones Biomédicas “Dr. Francisco J. Triana Alonso”, Facultad de Ciencias de la Salud, Universidad de Carabobo. Maracay, Venezuela. Escuela de Bioanálisis, Facultad de Ciencias de la Salud, Universidad de Carabobo. Maracay, Venezuela. Licenciada en Bioanálisis; Rivera, María G.; Instituto de Investigaciones Biomédicas “Dr. Francisco J. Triana Alonso”, Facultad de Ciencias de la Salud, Universidad de Carabobo. Maracay, Venezuela. Licenciada en Bioanálisis.; Viettri, Mercedes; Biomédicas “Dr. Francisco J. Triana Alonso”, Facultad de Ciencias de la Salud, Universidad de Carabobo. Maracay, Venezuela. Licenciada en Bioanálisis.; Lares, María; Instituto de Investigaciones Biomédicas “Dr. Francisco J. Triana Alonso”, Facultad de Ciencias de la Salud, Universidad de Carabobo. Maracay, Venezuela. Técnico superior universitario en Química.; Morocoima, Antonio; Centro de Medicina Tropical de Oriente, Facultad de Medicina, Universidad de Oriente. Anzoátegui, Venezuela. Médico cirujano, magíster en Parasitología.; Herrera, Leidi; Instituto de Zoología y Ecología Tropical, Facultad de Ciencias, Universidad Central de Venezuela. Caracas, Venezuela. Biólogo, doctora en Ciencias.; Ferrer, Elizabeth; Instituto de Investigaciones Biomédicas “Dr. Francisco J. Triana Alonso”, Facultad de Ciencias de la Salud, Universidad de Carabobo. Maracay, Venezuela. Departamento de Parasitología, Facultad de Ciencias de la Salud, Universidad de Carabobo. Maracay, Venezuela. Licenciada en bioanálisis, doctora en Biología Molecular.

    2014-01-01

    Objectives. To compare two extraction protocols of Trypanosoma cruzi DNA for use in DNA amplification of kinetoplast minicircles (kDNA) through the technique of Polymerase Chain Reaction (PCR). Materials and methods. Epimastigotes of T. cruzi were cultured in axenic conditions and masses from 1.5 to 100 x 106 parasites were obtained. DNA extraction was performed using two protocols: extraction with organic solvents (phenol/chloroform), and with resin (Chelex®100), from different parasitic sed...

  19. Optimisation of storage conditions for maintaining culturability of penicillin-susceptible and penicillin-resistant isolates of Streptococcus pneumoniae in transport medium.

    Science.gov (United States)

    Mason, C K; Goldsmith, C E; Moore, J E; McCarron, P; Leggett, P; Montgomery, J; Coulter, W A

    2010-01-01

    Methods employed by the World Health Organization (WHO) are used during this study to determine the optimum storage conditions for maintaining the culturability of Streptococcus pneumoniae in skimmed milk, tryptone, glucose and glycerin (STGG) transport medium. A comparison of S. pneumoniae strains sensitive and resistant to penicillin showed no significant difference in their survival ability in STGG medium. Furthermore, it is confirmed that storage at -70 degrees C remains most effective for maintaining viability by culture of S. pneumoniae. Storage at -20 degrees C would only be acceptable in the short-term, while storage at +4 degrees C is not recommended. Of note, this study has shown STGG medium at room temperature to be an efficient growth medium for pneumococci in the short-term. This work will help to establish robust sampling protocols when performing community studies to ensure culturability of comparison between community and laboratory pneumococci survival.

  20. Differentiation of mouse iPS cells into ameloblast-like cells in cultures using medium conditioned by epithelial cell rests of Malassez and gelatin-coated dishes.

    Science.gov (United States)

    Yoshida, Koki; Sato, Jun; Takai, Rie; Uehara, Osamu; Kurashige, Yoshihito; Nishimura, Michiko; Chiba, Itsuo; Saitoh, Masato; Abiko, Yoshihiro

    2015-09-01

    Induced pluripotent stem (iPS) cells are generated from adult cells and are potentially of great value in regenerative medicine. Recently, it was shown that iPS cells can differentiate into ameloblast-like cells in cultures using feeder cells. In the present study, we sought to induce differentiation of ameloblast-like cells from iPS cells under feeder-free conditions using medium conditioned by cultured epithelial cell rests of Malassez (ERM) cells and gelatin-coated dishes. Two culture conditions were compared: co-cultures of iPS cells and ERM cells; and, culture of iPS cells in ERM cell-conditioned medium. Differentiation of ameloblast-like cells in the cultures was assessed using real-time RT-PCR assays of expression of the marker genes keratin 14, amelogenin, and ameloblastin and by immunocytochemical staining for amelogenin. We found greater evidence of ameloblast-like cell differentiation in the cultures using the conditioned medium. In the latter, the level of amelogenin expression increased daily and was significantly higher than controls on the 7th, 10th, and 14th days. Expression of ameloblastin also increased daily and was significantly higher than controls on the 14th day. The present study demonstrates that mouse iPS cells can be induced to differentiate into ameloblast-like cells in feeder-free cell cultures using ERM cell-conditioned medium and gelatin-coated dishes.

  1. Multiwavelength Resonance Raman Characterization of the Effect of Growth Phase and Culture Medium on Bacteria.

    Science.gov (United States)

    Kunapareddy, Nagapratima; Grun, Jacob; Lunsford, Robert; Nikitin, Sergei; Wang, Zheng; Gillis, David

    2015-08-01

    We examine the use of multiwavelength ultraviolet (UV) resonance-Raman signatures to identify the effects of growth phase and growth medium on gram-positive and gram-negative bacteria. Escherichia coli (E. coli), Citrobacter koseri (C. koseri), Citrobacter braakii (C. braakii), and Bacillus cereus (B. cereus) were grown to logarithmic and stationary phases in nutrient broth and brain heart infusion broth. Resonance Raman spectra of bacteria were obtained at multiple wavelengths between 220 and 260 nm; a range that encompasses the resonance frequencies of cellular constituents. We find that spectra of the same bacterial species exhibit differences due to both growth condition and growth phase, but the larger differences reflect changes due to growth phase. The differences in the Raman spectra correlate with genetic differences among the species. Using a Pearson correlation based algorithm, we achieve successful identification of these bacteria in 83% of the cases.

  2. [Effects of medium-weight molecules isolated from plasma of healthy and burnt animals on the cellular composition of cultured bone marrow erythroblastic islets].

    Science.gov (United States)

    Volchegorskiĭ, I A; Tishevskaia, N V; Kuznetsov, D A

    2002-01-01

    The so-called median-weight molecular fractions (M, B = 0.5-1.5 kDa) exerting a marked hemopoietic effect in the cultured bone marrow erythroblastic islets were isolated from the plasma deproteinates of healthy dogs and animals with acute inflammation (thermal skin burn) by Sephadex G-15 gel chromatography. Acute inflammation is attended by the occurrence of blood medium-weight molecular components that suppress erythropiesis and by the concurrent high rate of granulocytopoiesis in the cultured bone marrow erythroblastic islets. Medium-weight molecular fractions from the blood of healthy dogs substantially were found to suppress erythropoiesis and to stimulate granulocytopoiesis in the cultured erythroblastic islets.

  3. Dio-sensimedia: a novel culture medium for rapid detection of extended spectrum β-lactamases

    Science.gov (United States)

    Cagatay, Atahan A; Kocagoz, Tanil; Eraksoy, Haluk

    2003-01-01

    Background Resistance to contemporary broad-spectrum β-lactams, mediated by extended-spectrum β-lactamases (ESBL), is an increasing problem worldwide. Many of the emerging antimicrobial resistance problems of this decade have been characterized by difficulty in the recognition of resistance in the laboratory, particularly by rapid susceptibility test methods. The plasmid-encoded ESBL represent such a resistance phenomenon that is difficult to recognize. We compared Dio-Sensimedia-ES (DSM-ES; Diomed, Istanbul, Turkey) and Mueller-Hinton (MH) agar in the double-disk synergy test (DDST) as a novel rapid system for detecting ESBL directly from bacterial culture. Methods Sixty ESBL-producing Klebsiella pneumoniae isolates cultured from blood (30), endotracheal aspirates (20), urine (5) and pus (5), as well as 40 Escherichia coli isolates cultured from endotracheal aspirates (15), urine (10), blood (8) and pus (7) were studied. Isolates positive for ESBL by the combined disk tests were tested with the DDST using MH and DSM-ES agar to detect ESBL-mediated resistance in K. pneumoniae and E. coli. DSM-ES agar was also used to determine the susceptibility of Enterobacteriaceae and staphylococci. Results Among 60 ESBL-producing K. pneumoniae isolates, 59 (98.3%) were identified as ESBL-positive by the DDST using MH, and 58 (96.6%), using DSM-ES agar. Of 40 ESBL-producing E. coli isolates, 38 (95%) were ESBL-positive by the DDST on MH agar, and 37 (92.5%), on DSM-ES agar. The average incubation period required for ESBL detection by the DDST on DSM-ES agar was 4 hours. Conclusions Since the DDST results were available within 4 hours when DSM-ES agar was used, the use of this media may significantly lower the length of hospital stay, the total cost for patient care and even the mortality rate by fascilitating early treatment against ESBL-producing organisms. PMID:14511397

  4. A linguistically level playing field: English-medium sport officiating between linguistic imperialism and cultural appropriation

    Directory of Open Access Journals (Sweden)

    Jacob Kornbeck

    2016-02-01

    Full Text Available This article revisits four problem areas of the use of English as a lingua franca by sports officials (including coaches, referees, etc. at international sporting events which were recently identified in an editorial by McNamee in the journal Sport, Ethics and Philosophy. These propositions are revisited by drawing on, inter alia, the theoretical models of ‘linguistic imperialism’ and ‘cultural appropriation’. The argument is made, in particular, that the ability of so-called ‘non-native’ users of English must not be underrated.

  5. Effect of culture medium on acid production from sorbitol by oral bacteria.

    Science.gov (United States)

    Kalfas, S; Edwardsson, S

    1990-08-01

    The fermentation of sorbitol or glucose and the acid production by strains belonging to the genera Actinomyces, Lactobacillus, and Streptococcus isolated from the predominant sorbitol-fermenting human dental plaque flora were studied in cultures in complex or defined bacteriologic broths and in saliva-based broth. The growth yields of Lactobacillus and Streptococcus in the saliva-based media and of Actinomyces in the defined broth were poor. Addition of fermentable carbohydrate to the saliva-based broth favored the growth of Streptococcus and Lactobacillus but not that of Actinomyces. The results showed obvious differences in the capacity of oral bacteria to ferment sorbitol between cultures in saliva-based and bacteriologic broths. Lactobacillus failed to utilize sorbitol when saliva was the only source of nutrients. Lower proportions of lactic and formic acids were formed from sorbitol by Actinomyces and Lactobacillus in the saliva-based than in the bacteriologic media. The findings illustrate some mechanisms possibly involved in the interactions between sorbitol and dental plaque flora.

  6. What is the DSM? Diagnostic manual, cultural icon, political battleground: an overview with suggestions for a critical research agenda

    OpenAIRE

    Greco, Monica

    2016-01-01

    The Diagnostic and Statistical Manual of Mental Disorders of the APA (known as the DSM) is a system for the classification of mental disorders that provides diagnostic criteria used by psychiatrists and experts in related fields. Although classification systems and standards are ubiquitous in social life, they are rarely conspicuous and almost never become an object of public debate. Yet the DSM has attained the status of a ‘cultural icon’ and has been an object of commentary and controversy ...

  7. Optimization of Production Medium for Mass Culture of Metarhizium anisopliae ARS 2231

    Energy Technology Data Exchange (ETDEWEB)

    Kang, Seung Woo [Korea University, Seoul (Korea); Moon, Ki Hyuk [Kyungwon University Technology Complex, Songnam (Korea); Yoon, Jeong Weon [Suwon University, Suwon (Korea)

    1999-11-01

    Chemical pesticides which have been commonly used for crop protection have many problems such as high production cost and environmental hazards. Entomogeneous fungi which attack living insects are very powerful means for microbiological insecticide. The purpose of this study is to investigate the culture conditions for mass production of Metarhizium anisopliae ARS 2231 which is a potential microbiological pesticide. The temperature and pH range for optimal cultivation were 28 deg. C and pH 5.0-7.0, respectively. For M. anisopliae ARS 2231, 3.0% (w/v) rice bran, 0.8% (w/v) hydrolyzed casein, and 0.3% (w/v) K{sub 2}HPO{sub 4} were found as the proper nutrients, considering cell mass, enzyme activities, and spore concentration. 16 refs., 10 tabs.

  8. Cell types and their status in Chlamydomonas-like algae (Chlorophyceae on agar medium culture

    Directory of Open Access Journals (Sweden)

    M.М. Pavlovska

    2014-04-01

    Full Text Available The classification of cell types under agar culture was proposed. Six cell morphotypes were allocated. The statuses were identified depending on the reduction of monade attributes of cells. The variants of transition from one cell morphotype to another under dissolving mucilage were shown. The monade, cocciod, palmeloid and gloeocysta morphotypes approximately equally represented in all clades. The asterococcus and mucogleocysta morphotypes presented only in Reinhardtinia аnd Oogamochlamydinia clades. Any morphotype isn’t typical for all clades of Chlamydomonas-like algae at once. The most of morphotypes numbers (5 from 6 are presented in Reinhardtinia clade. This demonstrates the diversity of the Reinhardtinia clade species. There are only one morphotype presented in Polytominia and Monadinia clades. There are four morphotypes presented in Oogamochlamydinia clade, three – in Moewusinia, two morphotypes – in Chloromonadinia.

  9. Isomaltulose production using free cells: optimisation of a culture medium containing agricultural wastes and conversion in repeated-batch processes.

    Science.gov (United States)

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

    2007-04-01

    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.

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

    Directory of Open Access Journals (Sweden)

    Sidik Marsudi

    2010-10-01

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

  11. Cultured 3T3L1 adipocytes dispose of excess medium glucose as lactate under abundant oxygen availability

    Science.gov (United States)

    Sabater, David; Arriarán, Sofía; Romero, María Del Mar; Agnelli, Silvia; Remesar, Xavier; Fernández-López, José Antonio; Alemany, Marià

    2014-01-01

    White adipose tissue (WAT) produces lactate in significant amount from circulating glucose, especially in obesity;Under normoxia, 3T3L1 cells secrete large quantities of lactate to the medium, again at the expense of glucose and proportionally to its levels. Most of the glucose was converted to lactate with only part of it being used to synthesize fat. Cultured adipocytes were largely anaerobic, but this was not a Warburg-like process. It is speculated that the massive production of lactate, is a process of defense of the adipocyte, used to dispose of excess glucose. This way, the adipocyte exports glucose carbon (and reduces the problem of excess substrate availability) to the liver, but the process may be also a mechanism of short-term control of hyperglycemia. The in vivo data obtained from adipose tissue of male rats agree with this interpretation.

  12. Combined effects of low-level laser therapy and human bone marrow mesenchymal stem cell conditioned medium on viability of human dermal fibroblasts cultured in a high-glucose medium.

    Science.gov (United States)

    Hendudari, Farzane; Piryaei, Abbas; Hassani, Seyedeh-Nafiseh; Darbandi, Hasan; Bayat, Mohammad

    2016-05-01

    Low-level laser therapy (LLLT) exhibited biostimulatory effects on fibroblasts viability. Secretomes can be administered to culture mediums by using bone marrow mesenchymal stem cells conditioned medium (BM-MSCs CM). This study investigated the combined effects of LLLT and human bone marrow mesenchymal stem cell conditioned medium (hBM-MSCs CM) on the cellular viability of human dermal fibroblasts (HDFs), which was cultured in a high-glucose (HG) concentration medium. The HDFs were cultured either in a concentration of physiologic (normal) glucose (NG; 5.5 mM/l) or in HG media (15 mM/l) for 4 days. LLLT was performed with a continuous-wave helium-neon laser (632.8 nm, power density of 0.00185 W/cm(2) and energy densities of 0.5, 1, and 2 J/cm(2)). About 10% of hBM-MSCs CM was added to the HG HDF culture medium. The viability of HDFs was evaluated using dimethylthiazol-diphenyltetrazolium bromide (MTT) assay. A significantly higher cell viability was observed when laser of either 0.5 or 1 J/cm(2) was used to treat HG HDFs, compared to the control groups. The cellular viability of HG-treated HDFs was significantly lower compared to the LLLT + HG HDFs, hBM-MSCs CM-treated HG HDFs, and LLLT + hBM-MSCs CM-treated HG HDFs. In conclusion, hBM-MSCs CM or LLLT alone increased the survival of HG HDFs cells. However, the combination of hBM-MSCs CM and LLLT improved these results in comparison to the conditioned medium.

  13. Innovative use of Mucuna monosperma (Wight) callus cultures for continuous production of melanin by using statistically optimized biotransformation medium.

    Science.gov (United States)

    Inamdar, Shrirang; Joshi, Swati; Bapat, Vishwas; Jadhav, Jyoti

    2014-01-20

    Melanins are predominantly indolic polymers which are having extensive applications in cosmetics, agriculture and medicine. In the present study, optimization of nutritional parameters influencing melanin production by Mucuna monosperma callus cultures was attempted using the response surface methodology (RSM). Standardization of four factors was carried out using the Box-Behnken design. The optimized levels of factors predicted by the model include tyrosine 0.978gL(-1), pH 5.85, SDS 34.55mgL(-1)and copper sulphate 21.14mgL(-1) tyrosine, which resulted in highest melanin yield of 0.887gL(-1). The optimization of medium using RSM resulted in a 3.06-fold increase in the yield of melanin. The ANOVA analysis showed a significant R(2)-value (0.9995), model F-value (1917.72) and probability (0.0001), with insignificant lack of fit. Optimized medium was used in the laboratory scale column reactor for the continuous production of melanin. Uninterrupted flow column exhibited maximum melanin production rate of 250mgL(-1)h(-1) which is the highest value ever reported using plant as a biotransformation source. Melanin production was confirmed by spectrophotometric and chemical analysis. Thus, this study demonstrates the production of melanin by M. monosperma callus, using a laboratory scale column reactor.

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

    Science.gov (United States)

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

    2010-10-01

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

  15. Cell viability studies and operation in cellular culture medium of n-type organic field-effect transistors

    Science.gov (United States)

    Barra, M.; Viggiano, D.; Di Capua, R.; Di Girolamo, F.; Santoro, F.; Taglialatela, M.; Cassinese, A.

    2012-02-01

    The possibility of the fabrication of organic devices suitable to be applied in bio-sensing fields depends largely on the availability of organic compounds displaying robust electrical properties even in aqueous solutions and effective biocompatibility features. In this paper, we report about the good cellular biocompatibility and the electrical response stability in an ionic medium of n-type organic transistors based on the recently developed PDI-8CN2 oligomer. The biocompatibility has been tested by analyzing the adhesion and viability of two different cell lines, human epithelial HeLa cells and murine neuronal F11 cells, on PDI-8CN2 films grown by organic molecular beam deposition (OMBD) on SiO2 substrates. The effect of film thickness on cell attachment was also tested. Uncoated SiO2 substrates were used as control surfaces and sexithiophene (T6) as device testing control. Moreover, the possible toxicity of -CN groups of PDI-8CN2 was tested on HeLa cell cultures, using PDI-8 and T6 molecules as controls. Results showed that, although at high concentration these organic compounds are toxic in solution, if they are presented in form of film, cell lines can attach and grow on them. The electrical response stability of PDI-8CN2 transistors in a cellular culture medium characterized by high concentrations of ionic species has been also investigated. For this purpose, low-voltage operation devices with VGS ranging from -5 V to 5 V, able to strongly reduce the influence of Faradaic currents coming from the electrical operation in an highly ionic environment, have been fabricated on 35 nm thick SiO2 layers and electrically characterized. These results are useful to experimentally define the main critical issues to be further addressed for the fabrication of reliable bio-sensors based on organic transistors.

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

    Science.gov (United States)

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

    2011-04-01

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

  17. Comparative study on using carbon or nitrogen limited medium to culture white rot fungi for reactive brilliant red dye K-2BP decolotization under non-sterile conditions

    Institute of Scientific and Technical Information of China (English)

    GAO; DaWen; WEN; XiangHua; QIAN; Yi

    2007-01-01

    In order to explore ways for the application of white rot fungus in dye effluent treatment under non-sterile conditions, experiment on decolorization of reactive brilliant red was carried out, employing nitrogen-limited and carbon-limited medium with C/N ratio of 56/2.2 and 28/44 (in mmol/L), respectively. The results showed that the decolorization rate reached 92% while culturing white rot fungus with nitrogen-limited medium; however, the decolorization process ended in carbon-limited medium (n(C)/n(N) = 28/44) because of bacterial contamination. In addition, pH rose up to 9.31 after 4 d of decolorization, which was caused by bacterial contamination in the carbon-limited system. Therefore, it is concluded that nitrogen-limited medium can inhibit bacterial growth to some extent while carbon-limited medium is more easily contaminated by bacteria. Nitrogen-limited medium is more suitable in culture of white rot fungus for decolorization of reactive dye. Medium with the ability of inhibiting yeast growth should be developed by adjusting other components of nitrogen-limited medium.

  18. Serum-free human MSC medium supports consistency in human but not in equine adipose-derived multipotent mesenchymal stromal cell culture.

    Science.gov (United States)

    Schubert, Susanna; Brehm, Walter; Hillmann, Aline; Burk, Janina

    2017-09-19

    For clinical applications of multipotent mesenchymal stromal cells (MSCs), serum-free culture is preferable to standardize cell products and prevent contamination with pathogens. In contrast to human MSCs, knowledge on serum-free culture of large animal MSCs is limited, despite its relevance for preclinical studies and development of veterinary cellular therapeutics. This study aimed to evaluate the suitability of a commercially available serum-free human MSC medium for culturing equine adipose-derived MSCs in comparison with human adipose MSCs. Enzyme-free isolation by explant technique and expansion of equine and human cells in the serum-free medium were feasible. However, serum-free culture altered the morphology and complicated handling of equine MSCs, with cell aggregation and spontaneous detachment of multilayers, compared to culture in standard medium supplemented with fetal bovine serum. Furthermore, proliferation and the surface immunophenotype of equine cells were more variable compared to the controls and appeared to depend on the lot of the serum-free medium. Particularly the expression of CD90 was different between experimental groups (P cells found in equine MSC samples cultured in serum-free medium (5.21-83.40%) compared to standard medium (86.20-99.50%). Additionally, small subpopulations expressing MSC exclusion markers such as CD14 (0.28-11.60%), CD34 (0.00-9.87%), CD45 (0.35-10.50%), or MHCII (0.00-3.67%) were found in equine samples after serum-free culture. In contrast, human samples displayed a more consistent morphology and a consistent CD29(+) (98.60-99.90%), CD73(+) (94.60-98.40%), CD90(+) (99.60-99.90%), and CD105(+) (97.40-99.80%) immunophenotype after culture in serum-free medium. The obtained data demonstrate that the serum-free medium was suitable for human MSC culture but did not lead to entirely satisfactory results in equine MSCs. This underlines that requirements regarding serum-free culture conditions are species

  19. Human umbilical cord stem cell conditioned medium versus serum-free culture medium in the treatment of cryopreserved human ovarian tissues in in-vitro culture: a randomized controlled trial.

    Science.gov (United States)

    Jia, Yingxian; Shi, Xiaohan; Xie, Yidong; Xie, Xiaochuan; Wang, Yan; Li, Shangwei

    2017-06-24

    To reduce young female fertility loss, the in-vitro culture of cryopreserved ovarian cortical tissues (OCTs) is considered an effective approach without delaying treatment and undergoing stimulation medicine. However, ischemic damage and follicular loss during the in-vitro culture of OCTs are major technical challenges. Human umbilical cord stem cells (HUMSCs) and their conditioned medium (HUMSC-CM) have been considered to be potential resources for regeneration medicine because they secrete cytokines and enhance cell survival and function. The aim of this study was to determine whether HUMSC-CM improves the development of frozen-thawed in-vitro cultured ovarian tissues compared with a serum-free culture medium (SF-CM). The thawed OCTs (n = 68) were cultivated in HUMSC-CM and SF-CM in vitro for 8 days, and the ovarian tissues were processed and analyzed by a classical histological evaluation. The microvessel density (MVD) and apotosis detection during in-vitro culture of OCTs were also performed. A significant difference in the rate of morphologically normal primordial follicles in the HUMSC-CM group was observed compared to that in the SF-CM group (group C) from days 2 to 4 (day 2: group B 58.0 ± 2.45% vs group C 32.0 ± 5.83%, p = 0.002; day 3: group B 55.5 ± 4.20% vs group C 21.0 ± 9.80%, p = 0.048; day 4: group B 52.0 ± 4.08% vs group C 21.5 ± 8.19%, p = 0.019). The microvessel density (MVD) detection showed a time-dependent increase and peaked on day 4. There was a significant difference between groups B (49.33 ± 0.58) and C (24.33 ± 3.79) (p = 0.036). The percentage of apoptotic follicles in group B was lower than that in group C on day 1 (13.75 ± 2.50% vs 27.0 ± 10.10%, p = 0.003), day 5 (11.75 ± 1.50% vs 51.0 ± 10.5%, p = 0.019) and day 7 (15.0 ± 5.10% vs 46.5 ± 21.75%, p = 0.018). These data have provided the first experimental evidence of the effect of

  20. A Preliminary Observation on the Development of Mouse Embryos Co-cultured with Human Oviductal Tissue or Conditioned Medium in Vitro

    Institute of Scientific and Technical Information of China (English)

    钟瑜; 张春雪; 潘善培

    1994-01-01

    The Present investigation has been carried out to examine the effect of human oviductal tissue co-culture system on the development of mouse embryos in vitro.Two-cell embryos collected from superovulated mouse were co-cultured with human oviductal tissue suspended in Ham'd F10+10%Fetal Calf Serum(F10 FCS),or in oviductal tissue conditioned medium and F10FCS as control.The results showed that the proportion developed into blastocyst,proportion of hatched and the velocity of cmbryo development were higher in both tissue co-culture and conditioned medium as compared with F10 FCS control.Furthermore,the velocity and percentage of embryomic devetopmem were higher in co-culture with ampullary tissue or its conditioned medium than that of isthmus,the effects of co-culture and conditioned medium on embryo development had no significant difference.All the embryos obtained from two co-culture systems could cleave normally,This experimental observation indicated that human oviductal epithelium might secrete some factors to promote the embryonic development in vitro.

  1. Effects of culture medium compositions on antidiabetic activity and anticancer activity of marine endophitic bacteria isolated from sponge

    Science.gov (United States)

    Maryani, Faiza; Mulyani, Hani; Artanti, Nina; Udin, Linar Zalinar; Dewi, Rizna Triana; Hanafi, Muhammad; Murniasih, Tutik

    2017-01-01

    High diversity of Indonesia marine spesies and their ability in producing secondary metabolite that can be used as a drug candidate cause this fascinating topic need to explore. Most of marine organisms explored to discover drug is macroorganism whereas microorganism (such as Indonesia marine bacteria) is very limited. Therefore, in this report, antidiabetic and anticancer activity of Indonesia marine bacteria isolated from Sponges's extract have been studied. Bacteria strain 8.9 which are collection of Research Center for Oseanography, Indonesian Institute of Sciences were from Barrang Lompo Island, Makasar, Indonesia. Bacteria were cultured in different culture medium compositions (such as: different pH, source of glucose and water) for 48 hours on a shaker, then they were extracted with ethyl asetate. Extracts of bacteria were tested by DPPH method (antioxidant activity), alpha glucosidase inhibitory activity method (antidiabetic activity), and Alamar Blue assay (anticancer activity) at 200 ppm. According to result, extract of bacteria in pH 8.0 exhibited the greatest antioxidant (19.27% inhibition), antidiabetic (63.95% inhibition) and anticancer activity of T47D cell line (44.62% cell viability) compared to other extracts. However, effect of addition of sugar sources (such as: glucose, sucrose, and soluble starch) and effect of addition of water/sea water exhibited less influence on their bioactivities. In conclusion, Indonesia marine bacteria isolated from sponge have potential a source of bioactive compound in drug discovery field.

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

    Directory of Open Access Journals (Sweden)

    Fernanda Bovo

    2014-09-01

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

  3. Supplementation of culture medium with L-carnitine improves development and cryotolerance of bovine embryos produced in vitro.

    Science.gov (United States)

    Takahashi, Toshikiyo; Inaba, Yasushi; Somfai, Tamas; Kaneda, Masahiro; Geshi, Masaya; Nagai, Takashi; Manabe, Noboru

    2013-01-01

    High lipid content in embryos is associated with low freezing tolerance. This study assessed the effects of exogenous L-carnitine, an enhancer of lipid metabolism, on the in vitro development and freezing survival of bovine embryos. Also, effects on metabolic activity, reactive oxygen species (ROS) and apoptosis were investigated. Supplementation of embryo culture medium with 1.518 mM or 3.030 mM L-carnitine significantly increased the rates of zygote development to the blastocyst stage and blastocyst cell numbers whereas 6.072 mM of this compound did not improve embryo development. Survival rates after slow freezing of blastocysts were significantly higher when embryos were cultured in the presence of 1.518 mM or 3.030 mM L-carnitine compared with the control. A lower density of lipid droplets was detected in L-carnitine-treated blastocysts compared with the control. L-carnitine significantly reduced ROS levels in 2-cell embryos but did not reduce ROS levels at later stages. The apoptotic cell rate was not different between control and L-carnitine-treated blastocysts. L-carnitine significantly increased ATP levels in 2-cell embryos but not at the 8-cell or blastocyst stages. L-carnitine increased the expression of metabolism-related ATP6 and COX1 genes in blastocysts. In conclusion, L-carnitine supplementation enhanced lipid metabolism in embryos resulting in improved development and cryotolerance of bovine blastocysts produced in vitro.

  4. Seleccionando materiales adecuados cultural y linguisticamente: Sugerencias para los proveedores de servicios (Selecting Culturally and Linguistically Appropriate Materials: Suggestions for Service Providers). ERIC Digest.

    Science.gov (United States)

    Santos, Rosa Milagros; Reese, Debbie

    The population of the United States is growing more culturally diverse each year, and this diversity is clearly evident among families with young children; however, individuals who work in early childhood programs are not as diverse as those they serve. Moreover, many early childhood professionals have little preparation for working with families…

  5. Spanish-Hispanic Culture from A to V (Actualidades to Venezuela): 72 Spanish-Language Interdisciplinary Cultural Themes with Suggested Resource Materials and Activities.

    Science.gov (United States)

    Moore, Merriam M.

    This guidebook is intended for use by teachers of Spanish (FLES through college level) and by teachers in Spanish bilingual programs. It lists all of the Spanish-speaking countries and 72 Spanish-language cultural themes, such as "Actualidades" (Current Events), "Carreras y espanol comercial" (Careers and Commercial Spanish), "Deportes" (Sports),…

  6. Spanish-Hispanic Culture from A to V (Actualidades to Venezuela): 72 Spanish-Language Interdisciplinary Cultural Themes with Suggested Resource Materials and Activities.

    Science.gov (United States)

    Moore, Merriam M.

    This guidebook is intended for use by teachers of Spanish (FLES through college level) and by teachers in Spanish bilingual programs. It lists all of the Spanish-speaking countries and 72 Spanish-language cultural themes, such as "Actualidades" (Current Events), "Carreras y espanol comercial" (Careers and Commercial Spanish), "Deportes" (Sports),…

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

    Science.gov (United States)

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

    2014-05-30

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

  8. 不同无土固体基质对辣椒生长的影响%Effect of Culturing Vegetable Solid Medium on the Growth and Development of Capsicum annuum L. without Soil Medium

    Institute of Scientific and Technical Information of China (English)

    李明福

    2011-01-01

    [目的]探索辣椒对无土固体基质栽培的适应性.[方法]选用烟草育苗专用基质、荞壳、河砂3种无土固体基质,对无土栽培辣椒的生长发育进行研究.[结果]采用烟草育苗专用基质的辣椒株高、生物产量和根重都优于荞壳和砂,说明烟草育苗专用基质更有利于辣椒生长.[结论]在无土固体基质栽培辣椒生产中,以选用烟草育苗专用基质为最佳.%[ Objective] The research aimed to discuss the suitability of Capsicum annuum L.to the culture of soilless solid medium. [ Method ] Three kinds of soilless solid media (tobacco seedling special medium, buckwheat shell and river sand)were selected to study the growth and development of Capsicum annuum L. in soilless culture were studied. [ Result] The plant height, biomass and root weight of Capsicum annuum L. in tobacco seedling special medium were better than that of buckwheat shell and river sand ,which indicated that tobacco seedling special medium was more favorable for the growth of Capsicum annuum L. [ Conclusion ] In the production of Capsicum annuum L. in the culture of soilless solid media, tobacco seedling special medium was the best choice.

  9. 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

    2004-01-01

    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.

  10. In vitro growth and organogenesis of Prosopis farcta plantlets (Fabaceae, Mimosoideae) in culture medium supplemented with various concentrations of Ca++ and Na+.

    Science.gov (United States)

    Stambouli, S; Bouzid, S; Dutuit, P; Harzallah-Skhiri, Fethia

    2012-03-01

    The objective of this study was to vary the mineral composition of the culture medium of Prosopis farcta seedlings per addition of Na+ and Ca++ ions with the aim to identify the culture media which support the growth and/or the expression of the in vitro plant organogenesis. The Na+ and Ca++ ions were added in the culture medium in various concentrations by taking the Gamborg medium, in which macroelements were diluted 10 times, as the basic one. After two months of culture, parameters relating to the vegetative development of plant seedlings and to the various expressions of organogenesis were measured. Weak concentrations in sodium and calcium ions as well as a weak concentration in Ca++ (0.1 mM) with 50 mM in Na+ support the best vegetative development of the plantlets. The most important percentage of plant seedlings presenting a bud initiation was obtained on a medium containing 0.1 mM of Na+ and 50 mM of Ca++. Our study defined several media likely to support in vitro development of Prosopis farcta plantlets allowing the selection of salt tolerant plants or cellular lines. Some other media were chosen for improving micropropagation of the species without adding growth substances.

  11. Stereoselective determination of midodrine and desglymidodrine in culture medium: application to a biotransformation study employing endophytic fungi.

    Science.gov (United States)

    Barth, Thiago; Pupo, Mônica Tallarico; Borges, Keyller Bastos; Okano, Laura Tiemi; Bonato, Pierina Sueli

    2010-05-01

    A CE method was developed and validated for the stereoselective determination of midodrine and desglymidodrine in Czapek culture medium to be applied to a stereoselective biotransformation study employing endophytic fungi. The electrophoretic analyses were performed using an uncoated fused-silica capillary and 70 mmol/L sodium acetate buffer solution (pH 5.0) containing 30 mmol/L heptakis (2, 3, 6-tri-O-methyl)-beta-CD as running electrolyte. The applied voltage and temperature used were 15 kV and 15 degrees C, respectively. The UV detector was set at 200 nm. The sample preparation was carried out by liquid-liquid extraction using ethyl acetate as extractor solvent. The method was linear over the concentration range of 0.1-12 microg/mL for each enantiomer of midodrine and desglymidodrine (r> or =0.9975). Within-day and between-day precision and accuracy evaluated by RSDs and relative errors, respectively, were lower than 15% for all analytes. The method proved to be robust by a fractional factorial design evaluation. The validated method was used to assess the midodrine biotransformation to desglymidodrine by the fungus Phomopsis sp. (TD2), which biotransformed 1.1% of (-)-midodrine to (-)-desglymidodrine and 6.1% of (+)-midodrine to (+)-desglymidodrine.

  12. HPLC analysis of midodrine and desglymidodrine in culture medium: evaluation of static and shaken conditions on the biotransformation by fungi.

    Science.gov (United States)

    Barth, Thiago; Aleu, Josefina; Pupo, Mônica Tallarico; Bonato, Pierina Sueli; Collado, Isidro G

    2013-01-01

    A high-performance liquid chromatography (HPLC) method is presented for the simultaneous determination of midodrine and desglymidodrine (DMAE) in Czapek-Dox culture medium, to be used in biotransformation studies by fungi. The HPLC analysis was conducted using a Lichrospher 100 RP18 column, acetonitrile-40 mmol/L formic acid solution (60:40, v/v) as mobile phase, and ultraviolet detection at 290 nm. The sample preparation was conducted by liquid-liquid extraction using ethyl acetate as extractor solvent. The method was linear over the concentration range of 0.4-40.0 µg/mL for midodrine (r ≥ 0.9997) and DMAE (r ≥ 0.9998). Within-day and between-day precision and accuracy were evaluated by relative standard deviations (≤ 8.2%) and relative errors (-7.3 to 7.4%), respectively. The validated method was used to assess midodrine biotransformation by the fungi Papulaspora immersa Hotson SS13, Botrytis cinerea UCA 992 and Botrytis cinerea 2100 under static and shaken conditions. Under shaken conditions, the biotransformation of midodrine to DMAE was more efficient for all studied fungi, especially for the fungus Botrytis cinerea 2100, which converted 42.2% of midodrine to DMAE.

  13. PENICILLIN-STREPTOMYCIN IN THE CULTURE MEDIUM DURING IN VITRO MATURATION (IVM OF BOVINE OOCYTES AFFECTS NUCLEAR MATURATION AND SUBSEQUENT EMBRYO DEVELOPMENT

    Directory of Open Access Journals (Sweden)

    SHIRAZI A

    2001-01-01

    Full Text Available Introduction: Standard concentrations of antibiotics in culture media are thought to have no detectable toxic effects on the cultured cells. However, since antibiotics are biologically active substances, the possibility that they interfere to some extent with cellular processes occurring in the cultured cells can not always be totally excluded. This study, therefore, was conducted to assess whether the presence of penicllin-streptomycin (pen-strep during in vitro maturation (IVM of bovine cumulus oocyte complexes (COCs affect nuclear and cytoplasmic maturation and subsequent embryo development. Materials and Methods: Bovine COCs were matured at 39oC in a humidified atmosphere with 5 % CO2 in air for 24 h in: 1- culture medium M 199 supplemented with 10 % FCS (Fetal calf serum, 0.05 IU/ml rhFSH (recombinant human FSH and 100 units penicillin and 100 ?g streptomycin/ ml. 2- culture medium M 199 without FCS and rhFSH in the presence of pen-strep. Cultures without antibiotics served as control. Six series of experiments, each consisted of at least 3 replicates, were performed. Results: In vitro maturation in the presence of pen-strep in culture medium supplemented with FCS and rhFSH significantly (P<0.05 increased the percentage of MII oocytes, however, when the COCs were divided, on the basis of appearance of the cumulus investment, into bright and dark groups, this effect was less obvious in both types of COCs, 76% vs 72% in bright COCs (P= 0.149 or 83% vs 80% in dark COCs (P=0.296 in treated and control groups respectively. The percentage of oocytes with type III of cortical granules (CGs distribution was not affected in the presence of pen-strep. The COCs expansion after IVM was not affected by the presence of antibiotics in culture medium. The subsequent embryo development of IVM/IVF produced ova, which were exposed to pen-strep during IVM, was significantly (P<0.05 decreased with respect to blastocyst formation by day 9. In vitro maturation in

  14. Hydrogen photoproduction in green algae Chlamydomonas reinhardtii sustainable over 2 weeks with the original cell culture without supply of fresh cells nor exchange of the whole culture medium.

    Science.gov (United States)

    Yagi, Takafumi; Yamashita, Kyohei; Okada, Norihide; Isono, Takumi; Momose, Daisuke; Mineki, Shigeru; Tokunaga, Eiji

    2016-07-01

    Unicellular green algae Chlamydomonas reinhardtii are known to make hydrogen photoproduction under the anaerobic condition with water molecules as the hydrogen source. Since the hydrogen photoproduction occurs for a cell to circumvent crisis of its survival, it is only temporary. It is a challenge to realize persistent hydrogen production because the cells must withstand stressful conditions to survive with alternation of generations in the cell culture. In this paper, we have found a simple and cost-effective method to sustain the hydrogen production over 14 days in the original culture, without supply of fresh cells nor exchange of the culture medium. This is achieved for the cells under hydrogen production in a sulfur-deprived culture solution on the {anaerobic, intense light} condition in a desiccator, by periodically providing a short period of the recovery time (2 h) with a small amount of TAP(+S) supplied outside of the desiccator. As this operation is repeated, the response time of transition into hydrogen production (preparation time) is shortened and the rate of hydrogen production (build up time) is increased. The optimum states of these properties favorable to the hydrogen production are attained in a few days and stably sustained for more than 10 days. Since generations are alternated during this consecutive hydrogen production experiment, it is suggested that the improved hydrogen production properties are inherited to next generations without genetic mutation. The properties are reset only when the cells are placed on the {sulfur-sufficient, aerobic, moderate light} conditions for a long time (more than 1 day at least).

  15. The presence of c-erbB-2 gene product-related protein in culture medium conditioned by breast cancer cell line SK-BR-3

    Energy Technology Data Exchange (ETDEWEB)

    Alper, O.; Yamaguchi, K.; Hitomi, J.; Honda, S.; Matsushima, T.; Abe, K. (National Cancer Center Research Institute, Tokyo (Japan))

    1990-12-01

    The Mr 185,000 glycoprotein encoded by human c-erbB-2/neu/HER2 gene, termed c-erbB-2 gene product, shows a close structural similarity with epidermal growth factor receptor and is now regarded to be a growth factor receptor for an as yet unidentified ligand. Abundant c-erbB-2 mRNA was demonstrated by Northern blot studies in the human breast cancer cell line SK-BR-3. Cellular radiolabeling experiments followed by immunoprecipitation with three different anti-c-erbB-2 gene product antibodies, recognizing extracellular domain, kinase domain, and carboxyl-terminal portion, respectively, demonstrated the production of a large amount of c-erbB-2 gene product which had the capacity to be phosphorylated. Immunization of mice with concentrated culture medium conditioned by SK-BR-3 cells always generated antibodies against c-erbB-2 gene product, demonstrating that this culture medium contained substance(s) immunologically indistinguishable from c-erbB-2 gene product. This observation was supported by the successful development of a monoclonal antibody against c-erbB-2 gene product, GFD-OA-p185-1, by immunizing mice with this culture medium. The biochemical nature of the substance(s) present in the culture medium was further characterized. When the culture medium conditioned by (35S)cysteine-labeled SK-BR-3 cells was immunoprecipitated by three different anti-c-erbB-2 gene product antibodies, only the antibody recognizing extracellular domain precipitated the (35S)-labeled protein with a molecular weight of 110,000, namely p110. The newly developed monoclonal antibody also immunoprecipitated this protein.

  16. Impact of medium volume and oxygen concentration in the incubator on pericellular oxygen concentration and differentiation of murine chondrogenic cell culture.

    Science.gov (United States)

    Oze, Hiroki; Hirao, Makoto; Ebina, Kosuke; Shi, Kenrin; Kawato, Yoshitaka; Kaneshiro, Shoichi; Yoshikawa, Hideki; Hashimoto, Jun

    2012-02-01

    Previous studies have demonstrated that oxygen environment is an important determinate factor of cell phenotypes and differentiation, although factors which affect pericellular oxygen concentration (POC) in murine chondrogenic cell culture remain unidentified. Oxygen concentrations in vivo were measured in rabbit musculoskeletal tissues, which were by far hypoxic compared to 20% O(2) (ranging from 2.29 ± 1.16 to 4.36 ± 0.51%). Oxygen concentrations in murine chondrogenic cell (C3H10T1/2) culture medium were monitored in different oxygen concentrations (20% or 5%) in the incubator and in different medium volumes (3,700 or 7,400 μl) within 25-cm(2) flasks. Chondrogenic differentiation was assessed by glycosaminoglycan production with quantitative evaluation of Alcian blue staining in 12-well culture dishes. Expression of chondrogenic genes, aggrecan, and type II collagen α1, was examined by quantitative real-time polymerase chain reaction. Oxygen concentrations in medium decreased accordingly with the depth from medium surface, and POC at Day 6 was 18.99 ± 0.81% in 3,700-μl medium (1,480-μm depth) and 13.26 ± 0.23% in 7,400-μl medium (2,960-μm depth) at 20% O(2) in the incubator, which was 4.96 ± 0.08% (1,480-μm depth) and 2.83 ± 0.42% (2,960-μm depth) at 5% O(2), respectively. The differences of POC compared by medium volume were statistically significant (p = 0.0003 at 20% and p = 0.001 at 5%). Glycosaminoglycan production and aggrecan gene expression were most promoted when cultured in moderately low POC, 1,000 μl (2,960-μm depth) at 20% O(2) and 500 μl (1,480-μm depth) at 5% O(2) in 12-well culture dishes. We demonstrate that medium volume and oxygen concentration in the incubator affect not only POC but also chondrogenic differentiation.

  17. Human mesenchymal stem cells possess different biological characteristics but do not change their therapeutic potential when cultured in serum free medium.

    Science.gov (United States)

    Wang, Youwei; Wu, Hehe; Yang, Zhouxin; Chi, Ying; Meng, Lei; Mao, Aibin; Yan, Shulin; Hu, Shanshan; Zhang, Jianzhong; Zhang, Yun; Yu, Wenbo; Ma, Yue; Li, Tao; Cheng, Yan; Wang, Yongjuan; Wang, Shanshan; Liu, Jing; Han, Jingwen; Li, Caiyun; Liu, Li; Xu, Jian; Han, Zhi Bo; Han, Zhong Chao

    2014-12-04

    Mesenchymal stem cells (MSCs) are widely investigated in clinical researches to treat various diseases. Classic culture medium for MSCs, even for clinical use, contains fetal bovine serum. The serum-containing medium (SCM) seems a major obstacle for MSCs-related therapies due to the risk of contamination of infectious pathogens. Some studies showed that MSCs could be expanded in serum free medium (SFM); however, whether SFM would change the biological characteristics and safety issues of MSCs has not been well answered. Human umbilical cord mesenchymal stem cells (hUC-MSCs) were cultured in a chemical defined serum free medium. Growth, multipotency, surface antigen expression, telomerase, immunosuppressive ability, gene expression profile and genomic stability of hUC-MSCs cultured in SFM and SCM were analyzed and compared side by side. hUC-MSCs propagated more slowly and senesce ultimately in SFM. SFM-expanded hUC-MSCs were different from SCM-expanded hUC-MSCs in growth rate, telomerase, gene expression profile. However, SFM-expanded hUC-MSCs maintained multipotency and the profile of surface antigen which were used to define human MSCs. Both SFM- and SCM-expanded hUC-MSCs gained copy number variation (CNV) in long-term in vitro culture. hUC-MCSs could be expanded in SFM safely to obtain enough cells for clinical application, meeting the basic criteria for human mesenchymal stem cells. hUC-MSCs cultured in SFM were distinct from hUC-MSCs cultured in SCM, yet they remained therapeutic potentials for future regenerative medicine.

  18. Adaptation of a Commonly Used, Chemically Defined Medium for Human Embryonic Stem Cells to Stable Isotope Labeling with Amino Acids in Cell Culture

    DEFF Research Database (Denmark)

    Liberski, A. R.; Al-Noubi, M. N.; Rahman, Z. H.

    2013-01-01

    developed by Ludwig et al. and commercially available as mTeSR1 [mTeSR1 is a trade mark of WiCell (Madison, WI) licensed to STEMCELL Technologies (Vancouver, Canada)]. This medium, together with adjustments to the culturing protocol, facilitates reproducible labeling that is easily scalable to the protein......Metabolic labeling with stable isotopes is a prominent technique for comparative quantitative proteomics, and stable isotope labeling with amino acids in cell culture (SILAC) is the most commonly used approach. SILAC is, however, traditionally limited to simple tissue culture regimens and only...... rarely employed in the context of complex culturing conditions as those required for human embryonic stem cells (hESCs). Classic hESC culture is based on the use of mouse embryonic fibroblasts (MEFs) as a feeder layer, and as a result, possible xenogeneic contamination, contribution of unlabeled amino...

  19. Use of MRSD medium and the hydrophobic grid membrane filter technique to differentiate between pediococci and lactobacilli in fermented meat and starter cultures.

    Science.gov (United States)

    Holley, R A; Millard, G E

    1988-10-01

    Modifications of MRS medium were made by incorporation of 0.1 M L-arginine-HCl, 0.0025% phenol red, 100 IU polymyxin B sulfate, by deletion of meat extract, use of only 1.2% (w/v) glucose and increase of Mn2+ to 1000 ppm. In addition, adoption of the hydrophobic grid membrane filter (HGMF) system with 0.025% Fast Green FCF dye and adjustment of the agar medium to pH 5.5 gave MRSD (differential) medium. Incubation at 25 degrees C anaerobically under N2 or CO2 followed by a post-growth staining procedure involving use of 0.4% (w/v) bromocresol purple yielded conditions under which pediococci colonies were blue whereas homo- and heterofermentative lactobacilli were green in color. Under these conditions, 7 pediococci, 16 lactobacilli, and 18 commercial meat starter cultures were successfully analyzed by plate count to yield a differential assessment of the lactobacilli and pediococci present without interference from the 9 other genera tested. Streptococcus lactis and Leuconostoc spp. produced blue and green colonies, respectively, at 25 degrees C which might interference but these organisms are not present in significant numbers in fermented meats. Pediococcus parvulus and Streptococcus faecalis produced green and blue colonies, respectively, but their very poor growth at 25 degrees C prevented their interference. Use of the developed MRSD medium was described for enumeration of both pediococci and lactobacilli in starter cultures and in fermenting dry sausages to enable documentation of starter culture performance.

  20. Generation of HIV-1 Gag VLPs by transient transfection of HEK 293 suspension cell cultures using an optimized animal-derived component free medium.

    Science.gov (United States)

    Cervera, Laura; Gutiérrez-Granados, Sonia; Martínez, Marta; Blanco, Julià; Gòdia, Francesc; Segura, María Mercedes

    2013-07-20

    Virus-like particles (VLPs) offer great promise as candidates for new vaccine strategies. Large-scale approaches for the manufacturing of HIV-1 Gag VLPs have mainly focused on the use of the baculovirus expression system. In this work, the development and optimization of an HIV-1 Gag VLP production protocol by transient gene expression in mammalian cell suspension cultures is reported. To facilitate process optimization, a Gag-GFP fusion construct enabling the generation of fluorescent VLPs was used. The great majority of Gag-GFP present in cell culture supernatants was shown to be correctly assembled into virus-like particles of the expected size and morphology consistent with immature HIV-1 particles. Medium optimization was performed using design of experiments (DoE). Culture medium supplementation with non-animal derived components including recombinant proteins and lipids of synthetic or non-animal-derived origin resulted in improved HEK 293 cell growth and VLP production. The maximum cell density attained using the optimized Freestyle culture medium was 5.4×10(6)cells/mL in batch mode, almost double of that observed using the unsupplemented medium (2.9×10(6)cells/mL). Best production performance was attained when cells were transfected at mid-log phase (2-3×10(6)cells/mL) with medium exchange at the time of transfection using standard amounts of plasmid DNA and polyethylenimine. By using an optimized production protocol, VLP titers were increased 2.4-fold obtaining 2.8μg of Gag-GFP/mL or 2.7×10(9)VLPs/mL according to ELISA and nanoparticle tracking quantification analyses, respectively. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. Type 1 cannabinoid receptor ligands display functional selectivity in a cell culture model of striatal medium spiny projection neurons.

    Science.gov (United States)

    Laprairie, Robert B; Bagher, Amina M; Kelly, Melanie E M; Dupré, Denis J; Denovan-Wright, Eileen M

    2014-09-05

    Modulation of type 1 cannabinoid receptor (CB1) activity has been touted as a potential means of treating addiction, anxiety, depression, and neurodegeneration. Different agonists of CB1 are known to evoke varied responses in vivo. Functional selectivity is the ligand-specific activation of certain signal transduction pathways at a receptor that can signal through multiple pathways. To understand cannabinoid-specific functional selectivity, different groups have examined the effect of individual cannabinoids on various signaling pathways in heterologous expression systems. In the current study, we compared the functional selectivity of six cannabinoids, including two endocannabinoids (2-arachidonyl glycerol (2-AG) and anandamide (AEA)), two synthetic cannabinoids (WIN55,212-2 and CP55,940), and two phytocannabinoids (cannabidiol (CBD) and Δ(9)-tetrahydrocannabinol (THC)) on arrestin2-, Gα(i/o)-, Gβγ-, Gα(s)-, and Gα(q)-mediated intracellular signaling in the mouse STHdh(Q7/Q7) cell culture model of striatal medium spiny projection neurons that endogenously express CB1. In this system, 2-AG, THC, and CP55,940 were more potent mediators of arrestin2 recruitment than other cannabinoids tested. 2-AG, AEA, and WIN55,212-2, enhanced Gα(i/o) and Gβγ signaling, with 2-AG and AEA treatment leading to increased total CB1 levels. 2-AG, AEA, THC, and WIN55,212-2 also activated Gα(q)-dependent pathways. CP55,940 and CBD both signaled through Gα(s). CP55,940, but not CBD, activated downstream Gα(s) pathways via CB1 targets. THC and CP55,940 promoted CB1 internalization and decreased CB1 protein levels over an 18-h period. These data demonstrate that individual cannabinoids display functional selectivity at CB1 leading to activation of distinct signaling pathways. To effectively match cannabinoids with therapeutic goals, these compounds must be screened for their signaling bias.

  2. Evaluation of Urea-motility-indole medium for recognition and differentiation of Salmonella and Shigella species in stool cultures.

    OpenAIRE

    Rosa Fraile, M; Vega Aleman, D; Fernandez Gutierrez, C

    1980-01-01

    A semisolid urea-motility-indole medium designed for detection in Enterobacteriaceae of urease activity, motility, and indole production in one tube was prepared and evaluated. The formulation of the medium was similar to that of Christensen urea agar, but the agar concentration was 0.2%, and 1% tryptone was added. Results with 687 strains of Enterobacteriaceae were the same as those obtained with standard test media (98% overall agreement). The urea-motility-indole medium was also used in co...

  3. 海洋尖尾藻培养液的基本特征%Basic Features of the Culture Medium of Oxyrrhis marina

    Institute of Scientific and Technical Information of China (English)

    安鑫龙; 李雪梅; 李亚宁

    2012-01-01

    [目的]研究海洋尖尾藻培养液的基本特征.[方法]以亚心形扁藻(Platymoras subcordiformis)和米氏凯伦藻(Karernia mikimotoi)作为海洋尖尾藻(Oxyrrhis marina)的饵料进行室内静置培养.[结果]海洋尖尾藻静王培养过程中培养液颜色发生变化,先由饵料藻培养液的颜色变为淡粉红色至粉红色,最终变为无色.海洋尖尾藻培养过程中瓶底首先出现少量沉淀物,随着培养时间延长,沉淀物上附着的饵料藻开始逐渐增多然后慢慢减少直至消失.[结论]该研究获得了海洋尖尾藻培养液的基本牲征,为海洋尖尾藻生态学研究提供了参考.%[ Objective ] To study the basic features of the culture medium of Oxyrrhis marina. [ Method ] The Oxyrrhis marina was indoor cultured with the baits of Platymonas subcordiformis and Karenia mikimotoi. [Result] During the static culture process, the color of the culture medium of Oxyrrhis marina changed firstly from light pink to pink and finally to colorless. Some precipitate appeared at the bottle bottom, with the extending culture time, the bait algae attached to the precipitate gradually increased, then decreased and finally disappeared. [ Conclusion] The study provides references for the screening of the culture medium of Oxyrrhis marina and realizes its long-term culture.

  4. Recurrent somatic embryogenesis in long-term cultures of Gentiana lutea L. as a source for synthetic seed production for medium-term preservation

    Directory of Open Access Journals (Sweden)

    Holobiuc Irina

    2012-01-01

    Full Text Available Our aim was to establish an efficient and reproducible system for producing synthetic seeds from recurrent somatic embryogenesis in long-term cultures of Gentiana lutea L. This species is a vulnerable medicinal plant, protected both at the national and international levels, and is included in different Red Lists and Books. In vitro culture, as an alternative to classical methods of preservation, allows for the cyclic multiplication of plant material and short-, medium- and long-term preservation of tissue collections. Biotechnological approaches allow for maintenance of the plant material in a confined space and protection against biotic and abiotic factors. Somatic embryogenesis (SE is the most efficient way to regenerate plants, ensuring material for preservation and fundamental research. In our experiment, recurrent somatic embryogenesis was developed in long-term cultures in the presence of sugar alcohols (mannitol, sorbitol and in the absence of growth factors. This process proceeded at a high rate, with adventive somatic embryos being generated in a continuous process, followed by maturation, germination and development into plants. To follow the somatic embryogenesis process, histological samples were made. We used these embryogenic cultures for synthetic seed production and medium-term conservation. The viability of somatic embryos after moderate osmotic stress treatment was tested using TTC. Our methodology relied on the induction of somatic embryogenesis in the presence of auxins in the first cycle of in vitro cultures, long-term high embryogenic culture maintenance in the presence of sugar alcohols and synthetic seed production.

  5. Optimization of a feed medium for fed-batch culture of insect cells using a genetic algorithm

    NARCIS (Netherlands)

    Marteijn, R.C.L.; Jurrius, O.; Dhont, J.; Gooijer, de C.D.; Tramper, J.; Martens, D.E.

    2003-01-01

    Insect cells have been cultured for over 30 years, but their application is still hampered by low cell densities in batch fermentations and expensive culture media. With respect to the culture method, the fed-batch culture mode is often found to give the best yields. However, optimization of the

  6. A Newly Defined and Xeno-Free Culture Medium Supports Every-Other-Day Medium Replacement in the Generation and Long-Term Cultivation of Human Pluripotent Stem Cells.

    Science.gov (United States)

    Ahmadian Baghbaderani, Behnam; Tian, Xinghui; Scotty Cadet, Jean; Shah, Kevan; Walde, Amy; Tran, Huan; Kovarcik, Don Paul; Clarke, Diana; Fellner, Thomas

    2016-01-01

    Human pluripotent stem cells (hPSCs) present an unprecedented opportunity to advance human health by offering an alternative and renewable cell resource for cellular therapeutics and regenerative medicine. The present demand for high quality hPSCs for use in both research and clinical studies underscores the need to develop technologies that will simplify the cultivation process and control variability. Here we describe the development of a robust, defined and xeno-free hPSC medium that supports reliable propagation of hPSCs and generation of human induced pluripotent stem cells (hiPSCs) from multiple somatic cell types; long-term serial subculturing of hPSCs with every-other-day (EOD) medium replacement; and banking fully characterized hPSCs. The hPSCs cultured in this medium for over 40 passages are genetically stable, retain high expression levels of the pluripotency markers TRA-1-60, TRA-1-81, Oct-3/4 and SSEA-4, and readily differentiate into ectoderm, mesoderm and endoderm. Importantly, the medium plays an integral role in establishing a cGMP-compliant process for the manufacturing of hiPSCs that can be used for generation of clinically relevant cell types for cell replacement therapy applications.

  7. Application and evaluation of modified sheep blood chocolate culture medium%改良羊血巧克力培养基的应用与评价

    Institute of Scientific and Technical Information of China (English)

    傅石明; 宋月芹; 朱香梅

    2015-01-01

    目的:评价改良羊血巧克力培养基的质量与应用价值。方法将 ATCC 10211流感嗜血杆菌接种到改良前后两种培养基上,比较两种培养基上嗜血杆菌的平均生长指数(GI 值);对352份经筛选的合格痰标本进行流感嗜血杆菌检测,比较两种培养基的阳性分离率。结果传统羊血巧克力培养基 GI 值为(3.69±0.58),改良羊血巧克力培养基 GI 值为(15.08±1.34),两种培养基 GI 值差异有统计学意义(t =25.31,P <0.01)。352份痰标本在改良羊血巧克力培养基上流感嗜血杆菌检出41例,阳性检出率11.65%。352份痰标本在传统羊血巧克力培养基上流感嗜血杆菌检出18例,阳性检出率5.54%。流感嗜血杆菌在两种巧克力培养基上的分离率差异有统计学意义(χ2=21.04,P <0.05)。结论流感嗜血杆菌在改良羊血巧克力培养基上生长良好,菌落明显,极易识别,有助于临床标本中流感嗜血杆菌的检出。%Objective To evaluate the quality and application value of improved sheep blood chocolate medium.Methods The ATCC 10211 of Haemophilus influenzae was inoculated into the modified medium and unmodified medium,the average growth index(GI)of Haemophilus influenzae in two kinds of culture medium was compared.Based on 352 selected qualified sputum specimens for detection of Haemophilus influenzae,the positive isolation rate of medium was compared between the two groups.Results GI value of the traditional blood chocolate culture medium was (3.69 ±0.58),which was significantly lower than (15.08 ±1.34)of the improved sheep blood chocolate culture medium,the difference was significant (t =25.31,P <0.01 ).352 sputum specimens in the improved sheep blood chocolate culture medium,Haemophilus influenzae detected in 41 cases,the positive rate was 11.65%.And 352 sputum specimens in traditional sheep blood chocolate culture,Haemophilus influenzae

  8. Quantitative 'Omics Analyses of Medium Chain Length Polyhydroxyalkanaote Metabolism in Pseudomonas putida LS46 Cultured with Waste Glycerol and Waste Fatty Acids.

    Directory of Open Access Journals (Sweden)

    Jilagamazhi Fu

    Full Text Available Transcriptomes and proteomes of Pseudomonas putida LS46 cultured with biodiesel-derived waste glycerol or waste free fatty acids, as sole carbon sources, were compared under conditions that were either permissive or non-permissive for synthesis of medium chain length polyhydroxyalkanoates (mcl-PHA. The objectives of this study were to elucidate mechanisms that influence activation of biopolymer synthesis, intra-cellular accumulation, and monomer composition, and determine if these were physiologically specific to the carbon sources used for growth of P. putida LS46. Active mcl-PHA synthesis by P. putida LS46 was associated with high expression levels of key mcl-PHA biosynthesis genes and/or gene products including monomer-supplying proteins, PHA synthases, and granule-associated proteins. 'Omics data suggested that expression of these genes were regulated by different genetic mechanisms in P. putida LS46 cells in different physiological states, when cultured on the two waste carbon sources. Optimal polymer production by P. putida LS46 was primarily limited by less efficient glycerol metabolism during mcl-PHA synthesis on waste glycerol. Mapping the 'Omics data to the mcl-PHA biosynthetic pathway revealed significant variations in gene expression, primarily involved in: 1 glycerol transportation; 2 enzymatic reactions that recycle reducing equivalents and produce key mcl-PHA biosynthesis pathway intermediates (e.g. NADH/NADPH, acetyl-CoA. Active synthesis of mcl-PHAs was observed during exponential phase in cultures with waste free fatty acids, and was associated with the fatty acid beta-oxidation pathway. A putative Thioesterase in the beta-oxidation pathway that may regulate the level of fatty acid beta-oxidation intermediates, and thus carbon flux to mcl-PHA biosynthesis, was highly up-regulated. Finally, the data suggested that differences in expression of selected fatty acid metabolism and mcl-PHA monomer-supplying enzymes may play a role in

  9. Trafficking of calcium-permeable and calcium-impermeable AMPA receptors in nucleus accumbens medium spiny neurons co-cultured with prefrontal cortex neurons.

    Science.gov (United States)

    Werner, Craig T; Murray, Conor H; Reimers, Jeremy M; Chauhan, Niravkumar M; Woo, Kenneth K Y; Molla, Hanna M; Loweth, Jessica A; Wolf, Marina E

    2017-04-01

    AMPA receptor (AMPAR) transmission onto medium spiny neurons (MSNs) of the adult rat nucleus accumbens (NAc) is normally dominated by GluA2-containing, Ca(2+)-impermeable AMPAR (CI-AMPARs). However, GluA2-lacking, Ca(2+)-permeable AMPA receptors (CP-AMPARs) accumulate after prolonged withdrawal from extended-access cocaine self-administration and thereafter their activation is required for the intensified (incubated) cue-induced cocaine craving that characterizes prolonged withdrawal from such regimens. These findings suggest the existence of mechanisms in NAc MSNs that differentially regulate CI-AMPARs and CP-AMPARs. Here, we compared trafficking of GluA1A2 CI-AMPARs and homomeric GluA1 CP-AMPARs using immunocytochemical assays in cultured NAc MSNs plated with prefrontal cortical neurons to restore excitatory inputs. We began by evaluating constitutive internalization of surface receptors and found that this occurs more rapidly for CP-AMPARs. Next, we studied receptor insertion into the membrane; combined with past results, the present findings suggest that activation of protein kinase A accelerates insertion of both CP-AMPARs and CI-AMPARs. We also studied constitutive cycling (net loss of receptors from the membrane under conditions where internalization and recycling are both occurring). Interestingly, although CP-AMPARs exhibit faster constitutive internalization, they cycle at similar rates as CI-AMPARs, suggesting faster reinsertion of CP-AMPARs. In studies of synaptic scaling, long-term (24 h) activity blockade increased surface expression and cycling rates of CI-AMPARs but not CP-AMPARs, whereas long-term increases in activity produced more pronounced scaling down of CI-AMPARs than CP-AMPARs but did not alter receptor cycling. These findings can be used to evaluate and generate hypotheses regarding AMPAR plasticity in the rat NAc following cocaine exposure. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Cadmium biosorption by free and immobilised microorganisms cultivated in a liquid soil extract medium: effects of Cd, pH and techniques of culture.

    Science.gov (United States)

    Lebeau, T; Bagot, D; Jézéquel, K; Fabre, B

    2002-05-27

    Instead of soil clean-up, a process not very technically and economically suited to agricultural soil contaminated by heavy metals (with a low concentration of heavy metals but highly or potentially highly contaminated surfaces), the control of the transfer of cadmium from the soil to the crops may well be a convenient method. We tested the bacterium ZAN-044, the actinomycete R27 and a basidiomycete Fomitopsis pinicola isolated for their ability to biosorb Cd, in order to inoculate agricultural soils afterwards. We then compared the cadmium biosorption by viable microbial cells which were free or immobilised in alginate beads and incubated in a soil extract liquid medium at various pH values (5, 6 and 7) and cadmium concentrations (1 and 10 mg/l). The Cd concentration in the medium had the most important effect on the percentage of Cd biosorbed by the microorganisms, but the culture mode (free or immobilised cells) was not a side effect. In the case of F. pinicola and the actinomycete R27, the percentage of Cd biosorbed by free cells did not decrease when the Cd concentration in the medium increased (6-42% at the lowest Cd concentration to 11-48% at 10 mg Cd/l). On the other hand, with a low Cd concentration (1 mg Cd/l), the percentage of Cd biosorbed by the bacterium ZAN-044 was maximum (69%) at pH 7, while this bacterium did not grow at 10 mg Cd/l and it did not accumulate Cd. For the three micro-organisms tested, relatively low specific biosorptions of Cd were observed, when the microorganisms were cultivated with a soil extract medium ('poor' medium), comparatively to those with a 'rich' medium. Finally, the choice of microorganism for the inoculation of contaminated soils depends on the cadmium level in the medium and on the distribution of the metal between the biomass and the medium.

  11. Inhibition of Listeria monocytogenes and Escherichia coli O157:H7 in liquid broth medium and during processing of fermented sausage using autochthonous starter cultures.

    Science.gov (United States)

    Pragalaki, T; Bloukas, J G; Kotzekidou, P

    2013-11-01

    The antimicrobial effect of two autochthonous starter cultures of Lactobacillus sakei was evaluated in vitro (in liquid broth medium) and in situ assays. The inactivation of foodborne pathogens Listeria monocytogenes (serotype 4ab No 10) and Escherichia coli O157:H7 ATCC 43888 was investigated during the production of fermented sausage according to a typical Greek recipe using L. sakei strains as starter cultures. The inactivation kinetics were modeled using GInaFiT, a freeware tool to assess microbial survival curves. By the end of the ripening period, the inhibition of L. monocytogenes was significant in treatments with L. sakei 8416 and L. sakei 4413 compared to the control treatment. A 2.2-log reduction of the population of E. coli O157:H7 resulted from the autochthonous starter culture L. sakei 4413 during sausage processing. The use of the autochthonous starter cultures constitutes an additional improvement to the microbial safety by reducing foodborne pathogens.

  12. Screening Test of Mother Culture Medium Formula of Pleurotus ery ngii%杏鲍菇母种培养基配方筛选试验

    Institute of Scientific and Technical Information of China (English)

    钮梦洁; 郝涤非

    2013-01-01

    In order to screen cultivation plan of Pleurotus eryngii ,taking Pleurotus eryngii as test material ,the screening test of mother culture medium formula of pleurotus eryngii were carried out by orthogonal design . The results showed that PDA medium respectively added 30 g bran ,10 g corn meal ,5 g yeast powder and 10 g glucose ,and that had the best culture effect for Pleurotus eryngii .%为优化杏鲍菇培养方案,以杏鲍菇子实体为试材,采用正交设计进行了杏鲍菇母种培养基配方筛选试验。结果表明:在PDA培养基中添加麸皮30 g、玉米粉10 g、酵母粉5 g、葡萄糖10 g对杏鲍菇母种培养效果最好。

  13. Inhibitory effect of organic acid salts on spoilage flora in culture medium and cured cooked meat products under commercial manufacturing conditions.

    Science.gov (United States)

    Drosinos, Eleftherios H; Mataragas, Marios; Kampani, Aikaterini; Kritikos, Dimitrios; Metaxopoulos, Ioannis

    2006-05-01

    Lactobacillus curvatus, isolated from a spoiled vacuum-packaged 'pariza' type meat product, was used to inoculate modified MRS broth containing sodium lactate, sodium acetate and potassium sorbate in different concentrations, alone or in inter se combinations. Two commercial preparations (MIX 1 and MIX 2) were also used containing combinations of the above antimicrobials. Results from the preservatives addition to the culture medium showed highest antimicrobial activity in the case of the sodium lactate (2%, 3% or 4%), sodium acetate (0.5%) and potassium sorbate (0.15%) combination. Results from the preservatives addition to two types of thermally processed meats showed that sodium lactate and the combination of sodium lactate, sodium acetate and potassium sorbate were the most effective; extending the products shelf life an additional 10 days. Finally, MIX 1 and MIX 2 suppressed the lactic acid bacteria (LAB) growth in the culture medium but not in the final product.

  14. [Effect of pH on suppressing the growth of other bacteria and fungi in culturing Phanerochaete chrysosporium in liquid medium].

    Science.gov (United States)

    Gao, Da-wen; Wen, Xiang-hua; Zhou, Xiao-yan; Zeng, Yong-gang; Qian, Yi

    2005-11-01

    Effect of different pH value on suppressing the growth of other bacteria and fungi in culturing Phanerochaete chrysosporium in liquid medium under non-sterile were investigated in agitated Erlenmeyer flasks. Results showed that nitrogen-limited liquid medium with pH3.6 and pH4.4 were contaminated only by yeast fungi when the Phanerochaete chrysosporium was incubated with spore inoculation under non-sterile condition for one day; however, nitrogen-limited liquid medium with pH5.6 was contaminated not only by yeast, but also by bacteria. These contaminated yeast and bacteria reduced the dye decolorizing ability of Phanerochaete chrysosporium . If after the Phanerochaete chrysosporium was incubated under sterile condition for 5 days, it can decolorize over 70% of the reactive brilliant red K-2BP within 45 hours under non-sterile condition, and this removal rate was close to or even higher than that under sterile condition. Phanerochaete chrysosporium cultured in the liquid medium with pH4.4 have the best decolorizing effect under non-sterile condition, and can decolorize up to 80% of the reactive brilliant red K-2BP in 24 hours. In additions, it was observed that by using the Phanerochaete chrysosporium incubated in above nitrogen-limited liquid medium with different pH under sterile condition for 5 days, the system were also contaminated by the other bacteria and yeast during decolorizing reactive brilliant red K-2BP under non-sterile condition, but the amount of these bacteria and yeast in liquid medium were too little to influence the Phanerochaete chrysosporium decolorizing reactive brilliant red K-2BP. So that, when Phanerochaete chrysosporium was used to decolorize reactive dyes under non-sterile condition, the incubation of Phanerochaete chrysosporium must be operated under sterile condition in order to achieve the higher decolorization.

  15. Serum-free medium supplemented with high-concentration FGF2 for cell expansion culture of human ear chondrocytes promotes redifferentiation capacity.

    Science.gov (United States)

    Mandl, Erik W; van der Veen, Simone W; Verhaar, Jan A N; van Osch, Gerjo J V M

    2002-08-01

    For tissue engineering of autologous cartilage, cell expansion is needed to obtain the cell numbers required. Standard expansion media contain bovine serum. This has several disadvantages, that is, the risk of transmitting diseases and serum-batch variations. The aim of this study was to find a serum-free medium with at least the same potential to expand cell numbers as serum-containing media. Ear chondrocytes of three young children were expanded in either serum-containing medium (SCM; DMEM with 10% fetal calf serum) or serum-free medium (SFM; DMEM with ITS+) supplemented with 5 or 100 ng/mL fibroblast growth factor-2 (FGF2). To promote cell adherence onto the culture flask, the serum-free conditions were cultured with 10% serum for 1 day after each trypsinization. After the fourth passage, the chondrocytes were encapsuled in alginate beads and redifferentiated in a SFM (DMEM with ITS+, hydrocortisone, and L-ascorbic acid) supplemented with 10 ng/mL IGF-I and 10 ng/mL TGFbeta-2. Results showed that expansion in SFM with 100 ng/mL FGF2 was comparable to expansion in SCM. Redifferentiation with SFM with IGF-I and TGFbeta-2 showed high collagen type II expression and high GAG/DNA production regardless of which expansion medium had been used. However, chondrocytes expanded in SFM with 100 ng/mL FGF2 resulted in less positive cells for collagen type I and 11-fibrau (a fibroblast membrane marker). The present study shows that it is possible to use serum-free medium for tissue engineering of cartilage. Expansion of immature ear chondrocytes in SFM supplemented with high-concentration FGF2 resulted in high cell numbers, which in addition had better redifferentiation capacity than cells expanded in medium with 10% serum.

  16. 马拉色菌培养基的改良及应用%Improvement and application of culture medium for malassezia species

    Institute of Scientific and Technical Information of China (English)

    穰真; 崔凡; 李薇; 王有为

    2013-01-01

    目的 改良传统的Leeming&Notman培养基,提高马拉色菌的培养效率,并实际应用.方法 设立含1%橄榄油、2%橄榄油和4%橄榄油的培养基组.在培养第3、5、7日测量菌落直径,提取基因组DNA并检测浓度,扩增ITS1-2区比较PCR产物质量.结果 马拉色菌在含4%橄榄油的培养基上生长最为迅速,产出的基因组DNA量最大,扩增得到的ITS1-2区产物质量最佳.结论 改良的含4%橄榄油的Leeming&Notman培养基有利于提高马拉色菌科研工作的效率.%Objective To improve the efficiency of traditional Leeming&Notman culture medium for Malassezia species and put into practical application. Methods Three groups of culture mediums containing 1% ,2% or 4% of olive oil were established for a-nalysis. On the 3rd,5th and 7th day of culturing, the diameter of each colony was measured,the genomic DNA of Malassezia isolates was extracted for quantitative analysis,and the ITS1-2 region was amplified for quality evaluation. Results Malassezia isolates on 4% of olive oil medium grew most rapidly,yielded the greatest amount of genomic DNA,and guaranteed best PCR products of ITS1-2 region. Conclusion Modified culture medium containing 4% of olive oil for Malassezia species is likely to improve the efficiency of research work.

  17. Anlysis the feasibility of chocolate medium as culture medium of haemophilus influenzae%巧克力培养基作为流感嗜血杆菌药敏培养基的可行性探讨

    Institute of Scientific and Technical Information of China (English)

    罗宇鹏; 卢先雷; 张婷; 唐仕萍

    2014-01-01

    Objective To study the feasibility of chocolate(CHO)culture-medium for haemophilus influenzae culture,which could be replace the HTM medium as drug susceptibility of haemophilus influenzae.Reducing the workload,saving the cost of inspection.Methods Collect clinical specimens of haemophilus influenzae 49 strains in May to December,2012,With CHO culture-medium and HTM medium for antibacterial drug sensitive test,And HTM medium as the reference standard.Analysis of haemophilus influenzae antibacterial drug sensitive test results, calculate the very important error,error calculation,minor errors and concordance rate.Results The doing of 8 kinds of antimicrobial agents,AMP、CXM、SXT、CIP、CRO、IPM、AZM didn′t appear very important error,the important error was < 5%,The several kinds of antibiotics susceptibility test results could use CHO culture-medium instead of HTM mediums.TCY drug sensitive test appeared relatively high secondary error of 32.7%,So the drug sensitive test of TCY cann′t replace HTM medium by chocolate culture-medium.Conclusion CHO culture-medium couldn′t re-place the HTM medium for haemophilus influenzae todrug sensitivity test in vitro.%目的:巧克力(CHO)培养基作为流感嗜血杆菌药敏培养基可行性探讨,判断巧克力培养基能否代替有效期短需现用现配的嗜血杆菌属试验培养基(HTM 培养基)作为流感嗜血杆菌的药敏培养基,为各级医院减少工作量,节约检验成本。方法收集2012年5~12月临床标本分离的流感嗜血杆菌49株,分别用巧克力培养基和CLSI 推荐培养基 HTM 培养基进行抗菌药物敏感试验,并以 HTM 培养基为参考标准,分析巧克力培养基上流感嗜血杆菌的抗菌药物敏感试验(以下简称药敏试验)结果,计算极重要误差、重要误差、次要误差,一致率。结果在所做的8种抗菌药物中,药物氨苄西林(AMP)、头孢呋辛(CXM)、复方磺胺甲恶唑(SXT

  18. Chemically defined medium and Caenorhabditis elegans

    Science.gov (United States)

    Szewczyk, Nathaniel J.; Kozak, Elena; Conley, Catharine A.

    2003-01-01

    BACKGROUND: C. elegans has been established as a powerful genetic system. Use of a chemically defined medium (C. elegans Maintenance Medium (CeMM)) now allows standardization and systematic manipulation of the nutrients that animals receive. Liquid cultivation allows automated culturing and experimentation and should be of use in large-scale growth and screening of animals. RESULTS: We find that CeMM is versatile and culturing is simple. CeMM can be used in a solid or liquid state, it can be stored unused for at least a year, unattended actively growing cultures may be maintained longer than with standard techniques, and standard C. elegans protocols work well with animals grown in defined medium. We also find that there are caveats to using defined medium. Animals in defined medium grow more slowly than on standard medium, appear to display adaptation to the defined medium, and display altered growth rates as they change the composition of the defined medium. CONCLUSIONS: As was suggested with the introduction of C. elegans as a potential genetic system, use of defined medium with C. elegans should prove a powerful tool.

  19. B7-H4 expression is elevated in human U251 glioma stem-like cells and is inducible in monocytes cultured with U251 stem-like cell conditioned medium.

    Science.gov (United States)

    Mo, Lian-Jie; Ye, Hong-Xing; Mao, Ying; Yao, Yu; Zhang, Jian-Min

    2013-12-01

    Previous studies indicated that B7-H4, the youngest B7 family, negatively regulates T cell-mediated immunity and is significantly overexpressed in many human tumors. Tumor stem cells are purported to play a role in tumor renewal and resistance to radiation and chemotherapy. However, the link between B7-H4 and tumor stem cells is unclear. In this study, we investigated B7-H4 expression in the medium of human glioma U251 cell cultures. Immunofluorescence results showed that U251 cells cultured in serum-free medium (supplemented with 2% B27, 20 ng/mL epidermal growth factor, 20 ng/mL basic fibroblast growth factor) maintained stem-like cell characteristics, including expression of stem cell marker CD133 and the neural progenitor cell markers nestin and SOX2. In contrast, U251 cells cultured in serum-containing medium highly expressed differentiation marker glial fibrillary acidic protein. Flow cytometry analysis showed serum-free medium-cultured U251 cells expressed higher intracellular B7-H4 than serum-containing medium-cultured U251 cells (24%-35% vs. 8%-11%, P cells revealed moderate expression of B7-H4 after stimulation with conditioned medium from U251 cells cultured in serum-containing medium. Moreover, conditioned medium from U251 stem-like cells had a significant stimulation effect on B7-H4 expression compared with serum-containing conditioned medium (P cells, and conditioned medium from these cells more effectively induced monocytes to express B7-H4 than conditioned medium from U251 cells cultured in the presence of serum. Our results show that U251 stem-like cells may play a more crucial role in tumor immunoloregulation with high expression of B7-H4.

  20. 不同培养基及采花期对辣椒花药培养的影响%Effect of Different Culture Medium and Florescence on Anther Culture of Capsicum annuum L

    Institute of Scientific and Technical Information of China (English)

    付文婷; 韩世玉; 邢丹; 廖芳芳; 苏丹; 张爱民; 蓬桂华; 何建文

    2014-01-01

    以5种辣椒为试材,研究不同培养基、采花期和基因型对辣椒花药培养的影响。结果表明:辣椒花药培养中,MS 培养基中花药褐化率、污染率均最低,膨大率最高,分别为33·2%、17·5%和39·4%;花药在3种培养基中反应不同,愈伤组织诱导率大小表现为 B5=CP>MS,胚状体诱导率大小为 MS>CP>B5。以 MS 为基本培养基诱导5种辣椒材料的单倍体,均能诱导出愈伤组织,国塔106愈伤组织诱导率最高,为1·18%,单身如意最低,仅为0·24%,杂6、8024和辛香8号诱导率为0·52%~1·03%。其中,只有杂6、8024和辛香8号3种材料诱导出胚状体,胚状体诱导率分别为0·06%、1·25%和0·05%,单身如意和国塔106均未诱导出胚状体。说明在相同的诱导条件下,不同基因型辣椒的单倍体诱导率存在很大差异。在盛花期采样,辣椒花药培养胚状体和愈伤组织诱导率最高,分别为1·482%和1·840%。%Effect of different medium,florescence and genotypes on anther culture of five Capsicum annuum L..The results showed that the browning rates and contamination rate of anther were the lowest in MS medium with 33·2% and 39·4% respectively,the swell rate of anther was the highest in MS medium with 17·5%;the order of callus induction rate was B5=CP>MS,the order of embryo rate was MS>CP>B5.The callus of five pepper haploids were all induced in MS medium,the callus induction rate of Guota 106 was the highest with 1·18%,the callus induction rate of Danshenruyi was the lowest with 0·24%,the callus induction rates of Za 6,8024 and Xinxiang 8 were from 0·52% to 1·03%.Only embryoid of three genotypes(Za 6,8024 and Xinxiang 8)were induced among the five genotypes,the induction rate were 0·06%,1·25%and 0·05%,Guota 106 and Danshenruyi did not give birth to embryoid,suggesting that the haploid induction rate was obvious different among different genotypes of

  1. Statistical optimization of medium composition and culture condition for the production of recombinant antilipopolysaccharide factor of Eriocheir sinensis in Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    JIANG Shan; LIU Mei; WANG Baojie; JIANG Keyong; WANG Lei

    2011-01-01

    Anti-lipopolysaccharide factors (ALFs) are important antimicrobial peptides that are isolated from some aquatic species.In a previous study,we isolated ALF genes from Chinese mitten crab,Eriocheir sinensis.In this study,we optimized the production of a recombinant ALF by expressing E.sinensis ALF genes in Escherichia coli maintained in shake-flasks.In particular,we focused on optimization of both the medium composition and the culture condition.Various medium components were analyzed by the Plackett-Burman design,and two significant screened factors,(NH4)2SO4 and KH2PO4,were further optimized via the central composite design (CCD).Based on the CCD analysis,we investigated the induction start-up time,the isopropylthio-D-galactoside (IPTG) concentration,the post-induction time,and the temperature by response surface methodology.We found that the highest level of ALF fusion protein was achieved in the medium containing 1.89g/L (NH4)2SO4 and 3.18 g/L KH2PO4,with a cell optical density of 0.8 at 600 nm before induction,an IPTG concentration of 0.5 mmol/L,a post-induction temperature of 32.7℃,and a post-induction time of 4 h.Applying the whole optimization strategy using all optimal factors improved the target protein content from 6.1% (without optimization) to 13.2%.We further applied the optimized medium and conditions in high cell density cultivation,and determined that the soluble target protein constituted 10.5% of the total protein.Our identification of the economic medium composition,optimal culture conditions,and details of the fermentation process should facilitate the potential application of ALF for further research.

  2. High-level secretion of a recombinant protein to the culture medium with a Bacillus subtilis twin-arginine translocation system in Escherichia coli.

    Science.gov (United States)

    Albiniak, Anna M; Matos, Cristina F R O; Branston, Steven D; Freedman, Robert B; Keshavarz-Moore, Eli; Robinson, Colin

    2013-08-01

    The twin-arginine translocation (Tat) system transports folded proteins across the plasma membrane in bacteria, and heterologous proteins can be exported by this pathway if a Tat-type signal peptide is present at the N-terminus. The system thus has potential for biopharmaceutical production in Escherichia coli, where export to the periplasm is often a favoured approach. Previous studies have shown that E. coli cells can export high levels of protein by the Tat pathway, and the protein product accummulates almost exclusively in the periplasm. In this study, we analysed E. coli cells that express the Bacillus subtilis TatAdCd system in place of the native TatABC system. We show that a heterologous model protein, comprising the TorA signal peptide linked to green fluorescent protein (TorA-GFP), is efficiently exported by the TatAdCd system. However, whereas the GFP is exported initially to the periplasm during batch fermentation, the mature protein is increasingly found in the extracellular culture medium. By the end of a 16-h fermentation, ~ 90% of exported GFP is present in the medium as active mature protein. The total protein profiles of the medium and periplasm are essentially identical, confirming that the outer membrane becomes leaky during the fermentation process. The cells are otherwise intact, and there is no large-scale release of cytoplasmic contents. Export levels are relatively high, with ~ 0.35 g GFP·L⁻¹ culture present in the medium. This system thus offers a means of producing recombinant protein in E. coli and harvesting directly from the medium, with potential advantages in terms of ease of purification and downstream processing.

  3. 烟草疫霉分离及生长培养基的选择%Selection of Isolating and Culturing Medium for Phytophthora nicotianae

    Institute of Scientific and Technical Information of China (English)

    苏凯; 桑维钧; 张新强; 王慧

    2013-01-01

    Different base medium adding different combination of antibiotics were applied for isolation and culture of Phytophthora nicotianae.The results showed that selective medium with Vs base medium (100mL/L V8juice+0.02 g/L CaCO3+2 g/Lagar) adding 10 mg/mL ampicillin +5 mg/mL nysfungin+5 mg/mL carbendazim had the best isolation effect as the successful isolation rate was 100%,and the hypha grew well on this medium.After separation,the hypha could be cultured in Vs base medium as the hypha was white,tight and strong with big bacteria clone and strong growth potential.%采用不同的基本培养基添加抗生素组合对烟草疫霉(Phytophthora nicotianae)进行分离培养.结果表明,以V8培养基(100 mL/L Vs汁+0.02 g/L CaCO3+2 g/L洋菜)为基本培养基附加10 mg/mL氨苄青霉素+5 mg/mL制霉素+5 mg/mL多菌灵的选择培养基分离烟草疫霉的效果较好,分离成功率达100%,菌丝的生长状态也较好.菌丝分离后在V8基本培养基中培养,菌丝紧密、浓白、粗壮,菌落大、长势旺.

  4. Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli

    Science.gov (United States)

    Jiang, Shan; Liu, Mei; Wang, Baojie; Jiang, Keyong; Wang, Lei

    2011-11-01

    Anti-lipopolysaccharide factors (ALFs) are important antimicrobial peptides that are isolated from some aquatic species. In a previous study, we isolated ALF genes from Chinese mitten crab, Eriocheir sinensis. In this study, we optimized the production of a recombinant ALF by expressing E. sinensis ALF genes in Escherichia coli maintained in shake-flasks. In particular, we focused on optimization of both the medium composition and the culture condition. Various medium components were analyzed by the Plackett-Burman design, and two significant screened factors, (NH4)2SO4 and KH2PO4, were further optimized via the central composite design (CCD). Based on the CCD analysis, we investigated the induction start-up time, the isopropylthio-D-galactoside (IPTG) concentration, the post-induction time, and the temperature by response surface methodology. We found that the highest level of ALF fusion protein was achieved in the medium containing 1.89 g/L (NH4)2SO4 and 3.18 g/L KH2PO4, with a cell optical density of 0.8 at 600 nm before induction, an IPTG concentration of 0.5 mmol/L, a post-induction temperature of 32.7°C, and a post-induction time of 4 h. Applying the whole optimization strategy using all optimal factors improved the target protein content from 6.1% (without optimization) to 13.2%. We further applied the optimized medium and conditions in high cell density cultivation, and determined that the soluble target protein constituted 10.5% of the total protein. Our identification of the economic medium composition, optimal culture conditions, and details of the fermentation process should facilitate the potential application of ALF for further research.

  5. Analysis of Isolation and Culture of Ureaplasma urealyticum and Micoplasma hominis by Solid Medium and liquid Medium%解脲、人型支原体固体和液体培养基方法比较

    Institute of Scientific and Technical Information of China (English)

    王俊文; 王红敏; 高慧双

    2016-01-01

    Objective To explore the genitourinary tract mycoplasma culture reason of false positive and false negative result in test and improvement countermeasures, to study the clinical detection of ureaplasma type (Uu) and mycoplasma (Mh) training Method 208 cases of urinary tract swab with solid separation culture Uu and Mh directly, inoculation fluid medium as control at the same time. Observe and record 24 h, 48 h, 72 h solid medium grow colonies mycoplasma positive cases and liquid culture positive cases, and statistical methods to analyze the differences of two methods. Results Solid medium direct separation of Uu and Mh have identified accurately, selectivity and the obvious advantages of colony characteristics, compared with the liquid culture is more suitable for clinical test of mycoplasma.%目的:探讨泌尿生殖道支原体培养试验中假阳性、假阴性结果的原因及改进的对策,研究临床检测解脲(Uu)和人型支原体(Mh)培养的方法。方法:对208例泌尿生殖道拭子直接用固体培养分离Uu和Mh,同时接种液体培养基作为对照。观察并记录24 h、48 h、72h固体培养基长出支原体菌落的阳性例数和液体培养阳性例数,并用统计学方法分析两种方法的差异。结果:固体培养基直接分离培养结果与液体培养在不同时间段所得到的结果,阳性率无统计学差异,99%的菌落于24 h~48 h之间可辨认。结论:固体培养基直接分离Uu和Mh具有鉴别准确,选择性强和菌落特征明显的优点,与液体培养相比更适用于支原体的临床检测。

  6. Factors Constraining the Informatization of China' s Small and Medium-sized Enterprises and Suggestions for Related Policies%我国中小企业信息化制约因素及政策建议

    Institute of Scientific and Technical Information of China (English)

    郭东强; 林美容

    2001-01-01

    Compared with the large-sized enterprises, the small and medium-sized enterprises have the characteristics of small scale, low management level, fragile fund basis and low capacity to resist business risk and external impact. Starting with the problems faced by the small and medium-sized enterprises in modern competition and informatization process, this paper explores the concrete policies and measures the small and medium-sized enterprises should adopt in the informatization construction.

  7. Development and characterization of a clinically compliant xeno-free culture medium in good manufacturing practice for human multipotent mesenchymal stem cells.

    Science.gov (United States)

    Chase, Lucas G; Yang, Sufang; Zachar, Vladimir; Yang, Zheng; Lakshmipathy, Uma; Bradford, Jolene; Boucher, Shayne E; Vemuri, Mohan C

    2012-10-01

    Human multipotent mesenchymal stem cell (MSC) therapies are currently being tested in clinical trials for Crohn's disease, multiple sclerosis, graft-versus-host disease, type 1 diabetes, bone fractures, cartilage damage, and cardiac diseases. Despite remarkable progress in clinical trials, most applications still use traditional culture media containing fetal bovine serum or serum-free media that contain serum albumin, insulin, and transferrin. The ill-defined and variable nature of traditional culture media remains a challenge and has created a need for better defined xeno-free culture media to meet the regulatory and long-term safety requirements for cell-based therapies. We developed and tested a serum-free and xeno-free culture medium (SFM-XF) using human bone marrow- and adipose-derived MSCs by investigating primary cell isolation, multiple passage expansion, mesoderm differentiation, cellular phenotype, and gene expression analysis, which are critical for complying with translation to cell therapy. Human MSCs expanded in SFM-XF showed continual propagation, with an expected phenotype and differentiation potential to adipogenic, chondrogenic, and osteogenic lineages similar to that of MSCs expanded in traditional serum-containing culture medium (SCM). To monitor global gene expression, the transcriptomes of bone marrow-derived MSCs expanded in SFM-XF and SCM were compared, revealing relatively similar expression profiles. In addition, the SFM-XF supported the isolation and propagation of human MSCs from primary human marrow aspirates, ensuring that these methods and reagents are compatible for translation to therapy. The SFM-XF culture system allows better expansion and multipotentiality of MSCs and serves as a preferred alternative to serum-containing media for the production of large scale, functionally competent MSCs for future clinical applications.

  8. Optimization of a serum-free culture medium for mouse embryonic stem cells using design of experiments (DoE) methodology.

    Science.gov (United States)

    Knöspel, Fanny; Schindler, Rudolf K; Lübberstedt, Marc; Petzolt, Stephanie; Gerlach, Jörg C; Zeilinger, Katrin

    2010-12-01

    The in vitro culture behaviour of embryonic stem cells (ESC) is strongly influenced by the culture conditions. Current culture media for expansion of ESC contain some undefined substances. Considering potential clinical translation work with such cells, the use of defined media is desirable. We have used Design of Experiments (DoE) methods to investigate the composition of a serum-free chemically defined culture medium for expansion of mouse embryonic stem cells (mESC). Factor screening analysis according to Plackett-Burman revealed that insulin and leukaemia inhibitory factor (LIF) had a significant positive influence on the proliferation activity of the cells, while zinc and L: -cysteine reduced the cell growth. Further analysis using minimum run resolution IV (MinRes IV) design indicates that following factor adjustment LIF becomes the main factor for the survival and proliferation of mESC. In conclusion, DoE screening assays are applicable to develop and to refine culture media for stem cells and could also be employed to optimize culture media for human embryonic stem cells (hESC).

  9. Use of new strains of Rhodobacter sphaeroides and a modified simple culture medium to increase yield and facilitate purification of the reaction centre.

    Science.gov (United States)

    Jun, D; Saer, R G; Madden, J D; Beatty, J T

    2014-05-01

    A new gene expression system was developed in Rhodobacter sphaeroides, replacing a pRK415-based system used previously. The broad host-range IPTG-inducible plasmid pIND4 was used to create the plasmid pIND4-RC1 for expression of the puhA and pufQBALMX genes, encoding the reaction centre (RC) and light-harvesting complex 1 (LH1) proteins. The strain R. sphaeroides ΔRCLH was used to make a knockout of the rshI restriction endonuclease gene, enabling electroporation of DNA into the bacterium; a subsequent knockout of ppsR was made, creating the strain R. sphaeroides RCx lacking this oxygen-sensing repressor of the photosynthesis gene cluster. Using pIND4-RC1, LH1 levels were increased by a factor of about 8 over pRS1 per cell in cultures grown semi-aerobically. In addition, the ppsR knockout allowed for photosynthetic pigment-protein complex synthesis in the presence of high concentrations of molecular oxygen; here, LH1 levels per cell increased by 20 % when grown under high aeration conditions. A new medium (called RLB) is the E. coli medium LB supplemented with MgCl2 and CaCl2, which was found to increase growth rates and final cell culture densities, with an increase of 30 % of LH1 per cell detected in R. sphaeroides RCx(pIND4-RC1) grown in RLB versus LB medium. Furthermore, cell density was about three times greater in RLB compared to semi-aerobic conditions. The combination of all the modifications resulted in an increase of LH1 and RC per mL of culture volume by approximately 35-fold, and a decrease in the length of culture incubation time from about 5 days to ~36 h.

  10. A water-soluble extract from cultured medium of Ganoderma lucidum (Reishi) mycelia attenuates the small intestinal injury induced by anti-cancer drugs.

    Science.gov (United States)

    Kashimoto, Naoki; Ishii, Satomi; Myojin, Yuki; Ushijima, Mitsuyasu; Hayama, Minoru; Watanabe, Hiromitsu

    2010-01-01

    The present study investigated whether a water-soluble extract from the culture medium of Ganoderma lucidum (Reishi) mycelia (MAK) is able to protect the small intestine against damage induced by anti-cancer drugs. Six-week-old male B6C3F1/Crlj mice were fed a basal diet (MF) alone or with various doses of MAK or Agarics blazei Murrill (AGA) beginning one week before treatment with the anti-cancer drugs. Mice were sacrificed 3.5 days after injection of the anti-cancer drug, the small intestine was removed and tissue specimens were examined for the regeneration of small intestinal crypts. In experiment 1, the number of regenerative crypts after the administration of 5-fluorouracil (5FU) intravenously (250 mg/kg) or intraperitoneally (250 or 500 mg/kg) was compared after treatment with MAK or AGA. MAK protected against 5FU-induced small intestinal injury whereas AGA did not. In experiment 2, we investigated the protective effect of MAK against small intestinal injury induced by the anti-cancer drugs: UFT (tegafur with uracil; 1,000 mg/kg, orally), cisplatin (CDDP; 12.5 and 25 mg/kg, intraperitoneally), cyclophosphamide (CPA; 250 mg/kg, orally) and gefitinib (Iressa; 2,000 and 4,000 mg/kg, orally). UFT and CDDP decreased the number of regenerative crypts, but treatment with MAK attenuated the extent of UFT- or CDDP-induced small intestinal injury. CPA or Iressa plus MAK up-regulated crypt regeneration. The present results indicate that MAK ameliorates the small intestinal injury caused by several anti-cancer drugs, suggesting that MAK is a potential preventive agent against this common adverse effect of chemotherapy.

  11. Sporothrix schenckii Sensu Lato identification in fragments of skin lesion cultured in NNN medium for differential diagnosis of cutaneous leishmaniasis.

    Science.gov (United States)

    Antonio, Liliane de Fátima; Pimentel, Maria Inês Fernandes; Lyra, Marcelo Rosandiski; Madeira, Maria de Fátima; Miranda, Luciana de Freitas Campos; Paes, Rodrigo Almeida; Brito-Santos, Fábio; Carvalho, Maria Helena Galdino Figueredo; Schubach, Armando de Oliveira

    2017-02-01

    Eighty-nine patients with clinical suspicion of leishmaniasis were referred for differential diagnosis. Sporothrix schenckii sensu lato was isolated in Novy-MacNeal-Nicolle + Schneider media in 98% of 64 patients with final diagnosis of sporotrichosis. This medium may be suitable for diagnosis of sporotrichosis in areas where cutaneous leishmaniasis is also endemic.

  12. Acceleration of antimicrobial susceptibility testing of positive blood cultures by inoculation of Vitek 2 cards with briefly incubated solid medium cultures.

    Science.gov (United States)

    Idelevich, Evgeny A; Schüle, Isabel; Grünastel, Barbara; Wüllenweber, Jörg; Peters, Georg; Becker, Karsten

    2014-11-01

    Briefly incubated agar cultures from positive blood cultures were used for antimicrobial susceptibility testing (AST) by Vitek 2. The cultivation time until inoculation was 3.8 h for Gram-positive cocci and 2.4 h for Gram-negative rods. The error rates were low, providing early and reliable AST without additional time or cost expenditure.

  13. Optimization of culture medium for novel cell-associated tannase production from Bacillus massiliensis using response surface methodology.

    Science.gov (United States)

    Belur, Prasanna D; Goud, Rakesh; Goudar, Dinesh C

    2012-02-01

    Naturally immobilized tannase (tannin acyl hydrolase, E.C. 3.1.1.20) has many advantages, as it avoids the expensive and laborious operation of isolation, purification, and immobilization, plus it is highly stable in adverse pH and temperature. However, in the case of cell-associated enzymes, since the enzyme is associated with the biomass, separation of the pure biomass is necessary. However, tannic acid, a known inducer of tannase, forms insoluble complexes with media proteins, making it difficult to separate pure biomass. Therefore, this study optimizes the production of cell-associated tannase using a "protein-tannin complex" free media. An exploratory study was first conducted in shake-flasks to select the inducer, carbon source, and nitrogen sources. As a result it was found that gallic acid induces tannase synthesis, a tryptose broth gives higher biomass, and lactose supplementation is beneficial. The medium was then optimized using response surface methodology based on the full factorial central composite design in a 3 l bioreactor. A 2(3) factorial design augmented by 7 axial points (alpha = 1.682) and 2 replicates at the center point was implemented in 17 experiments. A mathematical model was also developed to show the effect of each medium component and their interactions on the production of cell-associated tannase. The validity of the proposed model was verified, and the optimized medium was shown to produce maximum cell-associated tannase activity of 9.65 U/l, which is 93.8% higher than the activity in the basal medium, after 12 h at pH 5.0, 30 degrees C. The optimum medium consists of 38 g/l lactose, 50 g/l tryptose, and 2.8 g/l gallic acid.

  14. Using tobacco waste extract in pre-culture medium to improve xylose utilization for l-lactic acid production from cellulosic waste by Rhizopus oryzae.

    Science.gov (United States)

    Zheng, Yuxi; Wang, Yuanliang; Zhang, Jianrong; Pan, Jun

    2016-10-01

    The aim of this work was to study the high-titer l-lactic acid production from cellulosic waste using Rhizopus oryzae. The tobacco waste water-extract (TWE) added with 5g/L glucose and 0.1g/L vitamin C was optimized as pre-culture medium for R. oryzae. Results found that compared to traditional pre-culture medium, it improved xylose consumption rate up to 2.12-fold and enhanced l-lactic acid yield up to 1.73-fold. The highest l-lactic acid concentration achieved was 173.5g/L, corresponding to volumetric productivity of 1.45g/Lh and yield of 0.860g/g total reducing sugar in fed-batch fermentation. This process achieves efficient production of polymer-grade l-lactic acid from cellulosic feedstocks, lowers the cost of fungal cell pre-culture and provides a novel way for re-utilization of tobacco waste.

  15. Potential of the propolis as storage medium to preserve the viability of cultured human periodontal ligament cells: an in vitro study.

    Science.gov (United States)

    Saxena, Payal; Pant, Vandana Aditya; Wadhwani, Kulvindar Kaur; Kashyap, Mahendra Pratap; Gupta, Saurabh Kumar; Pant, Aditya Bhushan

    2011-04-01

    In vitro experiments were carried out to evaluate the potential of propolis, a natural resin known for its wide therapeutic window, as storage medium to preserve the viability of cultured human periodontal ligament (PDL) cells. Primary cultures of human PDL cells were subjected to either independent exposure of propolis (2.5%, 5.0%, 10.0%, and 20.0%), Hank's balanced salt solution (HBSS), milk (0.5%), artificial saliva, Dulbecco's modified Eagle's medium (DMEM) or combination of propolis 10% + DMEM, propolis 20% + DMEM for 30 min to 24 h at 37 °C. Cell viability was assessed using standard endpoints i.e., tetrazolium bromide salt (MTT), neutral red uptake, and trypan blue dye exclusion assay. In general, combinations of propolis 10% + DMEM, propolis 20% + DMEM, and DMEM alone were found to be better than other media used in this study. The difference in the potentials of these media to maintain the cell viability reached to the statistically significant levels by 24 h, when compared with other media used viz., propolis 2.5% (P propolis 5.0% (P propolis 10.0% (P propolis 20.0% (P milk (P propolis 10% + DMEM, propolis 20% + DMEM, and DMEM alone are equally good as storage media of choice to keep PDL cells viable during extra-alveolar period up to 24 h. Other more readily available medium such as milk may serve as appropriate alternative storage medium for shorter time periods i.e., up to 12 h. © 2011 John Wiley & Sons A/S.

  16. Defined Essential 8™ Medium and Vitronectin Efficiently Support Scalable Xeno-Free Expansion of Human Induced Pluripotent Stem Cells in Stirred Microcarrier Culture Systems

    Science.gov (United States)

    Badenes, Sara M.; Fernandes, Tiago G.; Cordeiro, Cláudia S. M.; Boucher, Shayne; Kuninger, David; Vemuri, Mohan C.; Diogo, Maria Margarida; Cabral, Joaquim M. S.

    2016-01-01

    Human induced pluripotent stem (hiPS) cell culture using Essential 8™ xeno-free medium and the defined xeno-free matrix vitronectin was successfully implemented under adherent conditions. This matrix was able to support hiPS cell expansion either in coated plates or on polystyrene-coated microcarriers, while maintaining hiPS cell functionality and pluripotency. Importantly, scale-up of the microcarrier-based system was accomplished using a 50 mL spinner flask, under dynamic conditions. A three-level factorial design experiment was performed to identify optimal conditions in terms of a) initial cell density b) agitation speed, and c) to maximize cell yield in spinner flask cultures. A maximum cell yield of 3.5 is achieved by inoculating 55,000 cells/cm2 of microcarrier surface area and using 44 rpm, which generates a cell density of 1.4x106 cells/mL after 10 days of culture. After dynamic culture, hiPS cells maintained their typical morphology upon re-plating, exhibited pluripotency-associated marker expression as well as tri-lineage differentiation capability, which was verified by inducing their spontaneous differentiation through embryoid body formation, and subsequent downstream differentiation to specific lineages such as neural and cardiac fates was successfully accomplished. In conclusion, a scalable, robust and cost-effective xeno-free culture system was successfully developed and implemented for the scale-up production of hiPS cells. PMID:26999816

  17. Meio modificado de cultura para caracterização de Salmonella lactose positiva A modified culture medium for the characterization of positive lactose strains of Salmonella

    Directory of Open Access Journals (Sweden)

    Deise Pasetto Falcão

    1976-03-01

    Full Text Available Foi desenvolvido um meio modificado de cultura para isolamento e caracterização de enterobactérias, visando especialmente salmonelas fermentadoras da lactose. No chamado "Meio modificado" as colônias das duas estirpes de Salmonella (lactose positivas e lactose negativas apresentam a morfologia idêntica, o que não ocorre quando são empregados os meios rotineiros à base de lactose, para isolamento de enterobactérias. Esse meio é uma modificação do meio de Hektoen Enteric Agar, do qual retirou-se lactose e adicionou-se xilose e L-lisina. Foi verificado que há possibilidade de diferenciar-se os diversos grupos de enterobactérias, empregando um meio de cultura sem lactose e usando como sistema diferenciador xilose e L-lisina. O meio modificado foi também avaliado quantitativamente comparando o seu poder enriquecedor ou inibitório, ao dos meios de Hektoen Enteric Agar, Brilliant Green Agar e SS Agar para diferentes grupos de enterobactérias.A modified culture medium was developed for the purpose of isolating and characterizing the enterobacterias, giving special attention to the Salmonella strains that ferment lactose. In this "Modified Medium" the colonies of the two strains of Salmonella show a morphological similarity. This does not occur with other culture media of enterobacteria, in which the basic carbohydrate is lactose. The Modified Medium is a modification of the Enteric Agar in which the lactose was substituted by xilose and L-lisine. It was verified that there is a possibility of differentiating between the different groups of enterobacteria by using a culture medium with no lactose and using xilose L-lisina as a differentiating system. The Modified Medium was also evaluated quantitatively comparing its enriching or inhibiting power with the Hektoen Enteric Agar, Brilliant Green Agar and the SS Agar media in relation to the different groups of enterobacteria.

  18. 植物乳杆菌代谢产细菌素的培养基优化%Culture Medium Optimization of Lactobacillus Plantarum for Bacteriocin Production

    Institute of Scientific and Technical Information of China (English)

    佟世生; 解洛香; 徐乐; 胡涛; 刘萍

    2012-01-01

    In this paper, the culture medium of Lactobacillus plantarum for bacteriocin production was optimized. The results showed that the culture medium had an important effect on germ formation and production of bacteriocin, in which carbon source was significant Overall, 5% molasses, 0.5% yeast extract, 2% tryptone, 0.4% KH2PO4, 0.1% MgSO4-7H2O, 0.5% CaCO3, 0.05% MnSO4 and 0.3% Tween 80 were the optimal combination of culture medium.%本文对植物乳杆菌代谢产细菌素的培养基进行一系列优化.结果显示,不同培养基对菌株生长和细菌素产量有重要的影响,其中以碳源的影响最为显著.综合考虑,5%糖蜜、0.5%酵母膏、2%胰蛋白胨、0.4%KH2PO4、0.1%MgSO4·7H2O、0.5%CaCO3、0.05% MnSO4和0.3%吐温80是植物乳杆菌生长和代谢产细菌素的最优培养基组合.

  19. Create the innovative culture of small and medium-sized private enterprises%打造中小民营企业创新文化探讨

    Institute of Scientific and Technical Information of China (English)

    孙仁祥

    2013-01-01

      我国中小民营企业生命周期只有3.5年,其根本原因在于创新文化建设方面存在不足。本文认为企业文化是企业成长最持久的决定因素,中小民营企业只有打造创新文化,塑造以创新为核心的企业价值观,树立以人为本的人才观,加强技术创新管理制度,同时政府要充分利用法制、税收、金融等政策,积极营造有利于中小民营企业自主创新的环境,才能真正实现企业的可持续发展。%  The life cycle of small and medium-sized private enterprises in China is only 3.5 years, the fundamental reason lies in the shortage of the construction of innovation culture. This paper argues that the enterprise culture is the most lasting decisive factor of enterprise growth, only to create innovation culture of small and medium-sized private enterprises, to create innovation as the core values of enterprises, establish the people-oriented concept of talent, strengthen technological innovation management system, at the same time, the government should make full use of legal, tax, financial policy, and actively create independent innovation environment of small and medium-sized private enterprise, in order to realize the sustainable development of enterprises.

  20. Optimum culture medium composition for lipopeptide production by Bacillus subtilis using response surface model-based ant colony optimization

    Indian Academy of Sciences (India)

    J Satya Eswari; M Anand; C Venkateswarlu

    2016-01-01

    Central composite rotatable design (CCRD) of experiments was used to obtain data for Lipopeptide and Biomass concentrations from fermentation medium containing the following five components: glucose,monosodium glutamate, yeast extract,MgSO4·7H2O, and K2HPO4. Data was used to develop a second order regression response surface model (RSM) which was coupled with ant colony optimization (ACO) to optimize the media compositions so as to enhance the productivity of lipopeptide. The optimized media by ACO was found to yield 1.501 g/L of lipopeptide concentration which was much higher compared to 1.387 g/L predicted by Nelder–Mead optimization (NMO). The optimum from ACO was validated experimentally. RSM-based ACO is thus shown to be an effective tool for medium optimization of biosurfactant production.

  1. Exploration on relative quality standard of Corynebacterium diphtheriae culture medium%白喉杆菌培养基相关质控标准探讨

    Institute of Scientific and Technical Information of China (English)

    邓雪莲; 李应伟; 唐朝晖

    2013-01-01

    Objective To establish relative quality standards of Corynebacterium diphtheriae culture medium.Methods Corynebacterium diphtheriae culture media with ≥ 1.8 g/L and < 1.8 g/L ammonia nitrogen were prepared respectively to use for Corynebacterium diphtheriae culture in large fermtor.The statistical analysis of the culture results were made by t test,and the optimal culture medium was identified.Fifteen batches of Corynebacterium diphtheriae culture media were prepared according to the identified formula and used to large fermentor culture.Quantities of diphtheria toxin harvested by culture were detected,and correlativity between ammonia nitrogen contents in Corynebacterium diphtheriae culture medium and diphtheria toxin produced by Corynebacterium diphtheriae was analysed.Fifteen batches of 15%-20% maltose solutions were made up and fed into large fermentor as supplement during Corynebacterium diphtheriae culture.Quantities of diphtheria toxin harvested by culture were detected,and correlativity between concentrations of maltose solutions and diphtheria toxin produced by Corynebacterium diphtheriae was analysed.Results Quantities of diphtheria toxin harvested from Corynebacterium diphtheriae culture media with ≥ 1.8 g/L ammonia nitrogen were significantly higher than those harvested from Corynebacterium diphtheriae culture media with < 1.8 g/L ammonia nitrogen(t =0.5635,P < 0.05).Positive linear correlation (r =0.52) was showed between ammonia nitrogen contents in Corynebacterium diphtheriae culture medium and quantities of diphtheria toxin in ranges of identified ammonia nitrogen contents (1.8-2.0 g/L).When concentrations of maltose solutions ranged from 15% to 20%,negative linear correlation (r =-0.53) was showed between concentrations of maltose solutions and quantities of diphtheria toxin.Conclusion When ammonia nitrogen contents in Corynebacterium diphtheriae culture medium and concentrations of maltose solutions for feeding into large

  2. Induced Biosynthesis of resveratrol and the prenylated stilbenoids arachidan-1 and arachidan-3 in hairy root cultures of peanut: effects of culture medium and growth stage

    Science.gov (United States)

    The peanut plant has evolved specialized biosynthetic mechanisms that allowed resisting infection by producing diverse secondary metabolites. Among these unique compounds are the stilbenoids, which include resveratrol analogues. Our previous research demonstrated that peanut hairy root cultures prov...

  3. Tooth Tissue Engineering: The Importance of Blood Products as a Supplement in Tissue Culture Medium for Human Pulp Dental Stem Cells.

    Science.gov (United States)

    Pisciolaro, Ricardo Luiz; Duailibi, Monica Talarico; Novo, Neil Ferreira; Juliano, Yara; Pallos, Debora; Yelick, Pamela Crotty; Vacanti, Joseph Phillip; Ferreira, Lydia Masako; Duailibi, Silvio Eduardo

    2015-11-01

    One of the goals in using cells for tissue engineering (TE) and cell therapy consists of optimizing the medium for cell culture. The present study compares three different blood product supplements for improved cell proliferation and protection against DNA damage in cultured human dental pulp stem cells for tooth TE applications. Human cells from dental pulp were first characterized as adult stem cells (ectomesenchymal mixed origin) by flow cytometry. Next, four different cell culture conditions were tested: I, supplement-free; II, supplemented with fetal bovine serum; III, allogeneic human serum; and IV, autologous human serum. Cultured cells were then characterized for cell proliferation, mineralized nodule formation, and colony-forming units (CFU) capability. After 28 days in culture, the comet assay was performed to assess possible damage in cellular DNA. Our results revealed that Protocol IV achieved higher cell proliferation than Protocol I (p = 0.0112). Protocols II and III resulted in higher cell proliferation than Protocol I, but no statistical differences were found relative to Protocol IV. The comet assay revealed less cell damage in cells cultured using Protocol IV as compared to Protocols II and III. The damage percentage observed on Protocol II was significantly higher than all other protocols. CFUs capability was highest using Protocol IV (p = 0.0018) and III, respectively, and the highest degree of mineralization was observed using Protocol IV as compared to Protocols II and III. Protocol IV resulted in significantly improved cell proliferation, and no cell damage was observed. These results demonstrate that human blood product supplements can be used as feasible supplements for culturing adult human dental stem cells.

  4. Detection of microorganisms in culture medium through the neutron radiographic technique; Deteccao de microorganismos em meios de cultura pela tecnica de neutrongrafia

    Energy Technology Data Exchange (ETDEWEB)

    Wacha, Reinaldo

    1999-05-01

    The study aims to obtain a more effective and faster method for the detection of bacteria in several culture media, such as potable water and blood. After the process growth in the culture medium, separation and suspension in buffer solution based in boron, the bacteria are deposited in track detectors that are submitted to thermal neutron beams (neutron flux: 2,2 x 10{sup 5} n.cm{sup -2}.s{sup -1}), resulting from the channel J-9 of the Argonauta research reactor, from the Nuclear Engineering Institute, IEN/CNEN. The latent tracks arisen from the alpha particles proceeding from the reaction B ({eta}, {alpha}) Li and, after having been revealed, are analyzed by an optical microscope that allows to detect the existence of the bacteria. Afterwards, they were analyzed in a nanoscope which helps the identification of the tracks of the alpha particles. (author)

  5. Could Externalized St. Jude Medical Riata® Lead Be a Culture Medium of a Polymicrobial Endocarditis? A Clinical Case

    Directory of Open Access Journals (Sweden)

    Zefferino Palamà

    2017-01-01

    Full Text Available We report the case of a man affected by polymicrobial endocarditis developed on a St. Jude Medical Riata lead with a malfunction because of the outsourcing of conductors. The patient was treated with antibiotic targeted therapy and showed different bacteria at the blood cultures and then underwent transvenous leads extraction. Vegetations were highlighted on the caval, atrial, and ventricular tracts of the Riata lead, but the cultures were all negative. The externalization of Riata lead may cause the malfunction but it could also promote bacterial colonies and vegetations. In conclusion, looking for early signs of infection is mandatory during Riata leads follow-up checks.

  6. Evaluation of BD BBL CHROMagar Staph aureus medium using AOAC and ISO culture methods. Performance tested method 100503.

    Science.gov (United States)

    Ritter, Vicki; Kircher, Susan; Sturm, Krista; Warns, Patty; Dick, Nancy

    2009-01-01

    BBL CHROMagar Staph aureus (CSA) medium was evaluated internally and externally for the isolation and enumeration of Staphylococcus aureus in cooked roast beef, smoked salmon, and shell eggs. All food matrixes were processed according to the AOAC Official Method 975.55 and ISO 6888-1:1999. Bacterial counts of S. aureus were compared on CSA to the reference media, Baird-Parker, at low, medium, and high contamination levels. Colony counts were converted to log10 for statistical analysis. Based on the paired t-test and one-way analysis of variance, no statistical difference was noted with the CSA method compared to the AOAC Official Method for the recovery of S. aureus for all food types and contamination levels. Compared to the ISO reference method, no statistical difference was found with the CSA method for any food type or contamination level, with the exception of low-level smoked salmon. A statistical difference was seen in the internal testing with the low-level contaminated smoked salmon where CSA recovered more colonies. The external testing showed no statistical difference with smoked salmon at the low level. The correlation coefficients ranged from 92.6 to 99.4%, demonstrating good correlation for overall levels in all food types and methods. The sensitivity and specificity of the CSA method using known isolates was 100%. The results of this study demonstrate that CSA is an effective medium for the isolation, enumeration, and presumptive identification of S. aureus in cooked roast beef, smoked salmon, and shell eggs in 24 h using ISO and AOAC official methods.

  7. Optimum conditions for growth in liquid medium of Oscillatoria formosa Bory used as the principal food in laboratory culture of intermediate hosts for schistosomosis and fasciolosis

    Directory of Open Access Journals (Sweden)

    Ferreira Filipa M.

    2000-09-01

    Full Text Available The rearing of snails, intermediate hosts of Schistosoma haematobium, S. intercalatum, S. bovis and Fasciola hepatica is the first step to maintain the life cycle of these parasites in laboratory in order to have biological material for the different studies, namely on the systematic biology and immunodiagnostic of schistosomosis and fasciolosis. According to the traditional method, the alga Oscillatoria formosa Bory (Cyanobacteria, principal food source for the snails, was cultivated in soil extract (Sampaio Xavier et al., 1968. However, it was sometimes very difficult to find the proper soil extract and the material was also contaminated by protozoa and fungi. In our work, using a new medium having as a base the Mineral Medium II (modified from Hughes et al., 1958 we found that O. formosa had a better growth response than in the soil extract medium. Snails fed on O. formosa reached three times the size of others at the same age, and they also reached sex maturity earlier, having more egg-masses per snail and, in addition, the rate of survival as well as the number of generations per year under laboratory conditions significantly increased. This culture was also easier to perform, and the axenic conditions easier to maintain.

  8. Purification and characterization of three chitinases and one beta-1,3-glucanase accumulating in the medium of cell suspension cultures of barley (Hordeum vulgare L.)

    DEFF Research Database (Denmark)

    Kragh, K.M.; Jacobsen, S.; Dalgaard Mikkelsen, J.

    1991-01-01

    Three basic chitinases and one basic beta-1,3-glucanase were secreted into the medium when embryogenic cell suspensions of barley (Hordeum vulgare L.) cv. 'Igri' were cultured as undifferentiated aggregates in the presence of 2,4-D. The enzymes were purified by affinity and ion exchange...... chromatography. Two of the chitinases were identified as the previously described endochitinases T and C from barley grain. The third and novel chitinase, designated K, was the major basic chitinase in the medium constituting 4% of the soluble protein. Chitinase K was found to be a 33-kDa endochitinase with p...... chitinases from barley aleurone and barley, bean and potato leaves. The purified beta-1,3-glucanase with a molecular weight (MW) of 32 kDa and pI greater-than-or-equal-to 9.8 constituted 1% of the soluble protein in the medium. Based on similar MW, pI and amino acid composition as well as identical N...

  9. Nutrient metabolism and optimization method for bacterial culture medium%细菌培养基中营养物质的代谢及优化方法

    Institute of Scientific and Technical Information of China (English)

    杨明明(综述); 李国晏; 应莲芳(审校)

    2016-01-01

    细菌培养基是细菌体外生长的基质,其中营养物质是细菌生长、繁殖的基本成分。因此,探索培养基中营养物质的代谢,优化培养基配方,对细菌体外培养的研究及规模化生产有着重要的指导意义。现就细菌培养基分类、营养物质、细菌在营养物质中的代谢及培养基优化方法等方面予以总结。%The bacterial culture medium is an essential growth substrate on bacterial growth and reproduction in vitro . Thus, it has a great siginificance in development of bacterial growth in vitro and in a related large scale production by inves-tigation of nutrient metabolism and searching for optimization formulation for the medium . This article sumirizes a general introduction about classification , nutrient , bacterial metabolism and optimization method for the medium.

  10. Growth and consumption of L-malic acid in wine-like medium by acclimated and non-acclimated cultures of Patagonian Oenococcus oeni strains.

    Science.gov (United States)

    Bravo-Ferrada, Bárbara Mercedes; Hollmann, Axel; Brizuela, Natalia; La Hens, Danay Valdés; Tymczyszyn, Elizabeth; Semorile, Liliana

    2016-09-01

    Five Oenococcus oeni strains, selected from spontaneous malolactic fermentation (MLF) of Patagonic Pinot noir wine, were assessed for their use as MLF starter cultures. After the individual evaluation of tolerance to some stress conditions, usually found in wine (pH, ethanol, SO2, and lysozyme), the behavior of the strains was analyzed in MLO broth with 14 % ethanol and pH 3.5 in order to test for the synergistic effect of high ethanol level and low pH and, finally, in a wine-like medium. Although the five strains were able to grow in MLO broth under low pH and/or high ethanol, they must be acclimated to grow in a wine-like medium. Additionally, glycosidase and tannase activities were evaluated, showing differences among the strains. The potential of the strains to ferment citrate was tested and two of the five strains showed the ability to metabolize this substrate. We did not detect the presence of genes encoding histidine, tyrosine descarboxylase, and putrescine carbamoyltransferase. All the strains tested exhibited good growth capacity and ability to consume L-malic acid in a wine-like medium after cell acclimation, and each of them showed a particular enzyme profile, which might confer different organoleptic properties to the wine.

  11. Medium selection for exopolysaccharide and biomass production in submerged cultures of culinary-medicinal mushrooms from Turkey

    NARCIS (Netherlands)

    Kizilcik, M.; Yamaç, M.; Griensven, van L.J.L.D.

    2010-01-01

    The present study investigates the exopolysaccharide (EPS) and biomass production of 18 strains of 15 species of culinary-medicinal higher Basidiomycetes in submerged culture under four different media. Gloeophyllum abietinum and Schizophyllum commune produced the highest EPS and biomass amounts—3.8

  12. The Cultural Orientations of Entrepreneurs and Employees' Job Satisfaction: The Turkish Small and Medium Sized Enterprises (SMEs) Case

    Science.gov (United States)

    Yetim, Nalan; Yetim, Unsal

    2006-01-01

    The primary purpose of this study was to determine whether cultural orientations which were pervasive and salient in the society of SMEs' entrepreneurs predict employees' job satisfaction. Paternalism, collectivism, individualism, power distance, uncertainty avoidance were assessed as pervasive and salient attributes for Turkish society. Data were…

  13. Validation of Performance of Plastic versus Glass Bottles for Culturing Anaerobes from Blood in BacT/ALERT SN Medium

    OpenAIRE

    Mirrett, Stanley; Joyce, Maria J.; Reller, L. Barth

    2005-01-01

    To validate performance, we compared the new plastic BacT/ALERT (bioMérieux, Durham, NC) SN bottle to the current glass SN bottle with samples of blood obtained for culture from adults and found them comparable for both recovery and speed of detection of microorganisms. We conclude that the safety advantage of plastic bottles can be achieved without compromising performance.

  14. Validation of performance of plastic versus glass bottles for culturing anaerobes from blood in BacT/ALERT SN medium.

    Science.gov (United States)

    Mirrett, Stanley; Joyce, Maria J; Reller, L Barth

    2005-12-01

    To validate performance, we compared the new plastic BacT/ALERT (bioMérieux, Durham, NC) SN bottle to the current glass SN bottle with samples of blood obtained for culture from adults and found them comparable for both recovery and speed of detection of microorganisms. We conclude that the safety advantage of plastic bottles can be achieved without compromising performance.

  15. The Cultural Orientations of Entrepreneurs and Employees' Job Satisfaction: The Turkish Small and Medium Sized Enterprises (SMEs) Case

    Science.gov (United States)

    Yetim, Nalan; Yetim, Unsal

    2006-01-01

    The primary purpose of this study was to determine whether cultural orientations which were pervasive and salient in the society of SMEs' entrepreneurs predict employees' job satisfaction. Paternalism, collectivism, individualism, power distance, uncertainty avoidance were assessed as pervasive and salient attributes for Turkish society. Data were…

  16. A genotype of modified vaccinia Ankara (MVA) that facilitates replication in suspension cultures in chemically defined medium.

    Science.gov (United States)

    Jordan, Ingo; Horn, Deborah; John, Katrin; Sandig, Volker

    2013-01-21

    While vectored vaccines, based on hyperattenuated viruses, may lead to new treatment options against infectious diseases and certain cancers, they are also complex products and sometimes difficult to provide in sufficient amount and purity. To facilitate vaccine programs utilizing host-restricted poxviruses, we established avian suspension cell lines (CR and CR.pIX) and developed a robust, chemically defined, culturing process for production of this class of vectors. For one prominent member, modified vaccinia Ankara (MVA), we now describe a new strain that appears to replicate to greater yields of infectious units, especially in the cell-free supernatant of cultures in chemically defined media. The new strain was obtained by repeated passaging in CR suspension cultures and, consistent with reports on the exceptional genetic stability of MVA, sequencing of 135 kb of the viral genomic DNA revealed that only three structural proteins (A3L, A9L and A34R) each carry a single amino acid exchange (H639Y, K75E and D86Y, respectively). Host restriction in a plaque-purified isolate of the new genotype appears to be maintained in cell culture. Processing towards an injectable vaccine preparation may be simplified with this strain as a complete lysate, containing the main burden of host cell contaminants, may not be required anymore to obtain adequate yields.

  17. Putrescine production via the agmatine deiminase pathway increases the growth of Lactococcus lactis and causes the alkalinization of the culture medium.

    Science.gov (United States)

    del Rio, Beatriz; Linares, Daniel M; Ladero, Victor; Redruello, Begoña; Fernández, Maria; Martin, Maria Cruz; Alvarez, Miguel A

    2015-01-01

    Lactococcus lactis is the most important starter culture organism used in the dairy industry. Although L. lactis species have been awarded Qualified Presumption of Safety status by the European Food Safety Authority, and Generally Regarded as Safe status by the US Food and Drug Administration, some strains can produce the biogenic amine putrescine. One such strain is L. lactis subsp. cremoris CECT 8666 (formerly L. lactis subsp. cremoris GE2-14), which was isolated from Genestoso cheese. This strain catabolizes agmatine to putrescine via the agmatine deiminase (AGDI) pathway, which involves the production of ATP and two ammonium ions. The present work shows that the availability of agmatine and its metabolization to putrescine allows for greater bacterial growth (in a biphasic pattern) and causes the alkalinization of the culture medium in a dose-dependent manner. The construction of a mutant lacking the AGDI cluster (L. lactis CECT 8666 Δagdi) confirmed the latter's direct role in putrescine production, growth, and medium alkalinization. Alkalinization did not affect the putrescine production pattern and was not essential for increased bacterial growth.

  18. Survival of the North American strain of viral hemorrhagic septicemia virus (VHSV) in filtered seawater and seawater containing ovarian fluid, crude oil and serum-enriched culture medium

    Science.gov (United States)

    Kocan, R.M.; Hershberger, P.K.; Elder, N.E.

    2001-01-01

     The North American strain of viral hemorrhagic septicemia virus (NA-VHSV) could be recovered for up to 40 h in natural filtered seawater (27 ppt) with a 50% loss of infectivity after approximately 10 h at 15°C. Addition of 10 ppb North Slope crude oil to the seawater had no effect on virus survival. However, when various concentrations of teleost ovarian fluid were added to seawater, virus could be recovered after 72 h at 0.01% ovarian fluid and after 96 h at 1.0%. When cell culture medium supplemented with 10% fetal bovine serum was added to the seawater, 100% of the virus could be recovered for the first 15 d and 60% of the virus remained after 36 d. These findings quantify NA-VHSV infectivity in natural seawater and demonstrate that ovarian fluid, which occurs naturally during spawning events, significantly prolongs the survival and infectivity of the virus. The extended stabilization of virus in culture medium supplemented with serum allows for low titer field samples to be collected and transported in an unfrozen state without significant loss of virus titer.

  19. Increasing the Richness of Culturable Arsenic-Tolerant Bacteria from Theonella swinhoei by Addition of Sponge Skeleton to the Growth Medium.

    Science.gov (United States)

    Keren, Ray; Lavy, Adi; Ilan, Micha

    2016-05-01

    Theonella swinhoei is an arsenic hyper-accumulator sponge, harboring a multitude of associated bacteria. These bacteria reside in the mesohyl, the dense extracellular matrix of the sponge. Previous elemental analysis of separated cell fractions from the sponge had determined that arsenic is localized to the associated bacteria. Subsequently, sponge-associated arsenic-tolerant bacteria were isolated here and grouped into 15 operational taxonomic units (OTUs, 97% similarity). Both culture-dependent and culture-independent work had revealed that T. swinhoei harbors a highly diverse bacterial community. It was thus hypothesized the acclimation of bacteria in the presence of a sponge skeleton, better mimicking its natural environment, would increase the yield of isolation of sponge-associated bacteria. Using seven modularly designed media, 380 bacteria isolates were grown and grouped into 22 OTUs. Inclusion of sponge skeleton in the growth medium promoted bacterial growth in all seven media, accounting for 20 of the 22 identified OTUs (the other two in a medium without skeleton). Diversity and richness indices were calculated for each treatment or combination of treatments with shared growth parameters. Integrating data inherent in the modularly designed media with the ecological indices led to the formation of new hypotheses regarding the aeration conditions and expected arsenic form in situ. Both aerobic and anoxic conditions are expected to occur in the sponge (temporally and/or spatially). Arsenate is expected to be the dominant (or even the only) arsenic form in the sponge.

  20. 不同培养基对蝉花培养物核苷类成分的影响%Effect of different culture medium on nucleoside component content in culture of Cordyceps cicadae

    Institute of Scientific and Technical Information of China (English)

    张忠亮; 陈桃宝; 尹彬; 崔俏俏; 张霜霜; 王玉芹

    2016-01-01

    目的 建立高效液相色谱(HPLC)法同时检测蝉花培养物4种核苷类成分,分析不同斜面培养基及固体培养基对4种核苷类成分的影响.方法 建立4种核酸类成分——尿苷、鸟苷、腺苷、N6-(2-羟乙基)腺苷的HPLC检测方法,应用Waters Symmetry shield TMRP C18 (250 mm×4.6mm,5μm)色谱柱;流动相为水-甲醇,梯度洗脱;体积流量1.0 mL/min;柱温30℃;进样量10 μL;检测波长260 nm;外标法测定4种核苷类成分水甲.对包括假单胞菌选择培养基(PSA)、沙氏葡萄糖琼脂培养基(SDAY,1 000 mL中含葡萄糖量分别为20、30、40 g)在内的斜面培养基和包括江苏农垦(SCM)、烟农24 (YN-24)、烟农15 (YN-15)、济麦22 (JM-22)在内的固体培养基培养的蝉花培养物中4种核酸类成分水平进行测定,利用SPSS16.0统计软件对测定结果进行多重统计分析,验证并找出引起差异的原因.结果 建立的HPLC法,4种核苷类成分分离度良好,精密度、稳定性、重复性、加样回收率、耐用性均符合要求;4种不同的斜面培养基对尿苷成分基本无影响(P>0.05),PSA对腺苷、鸟苷、N6-(2-羟乙基)腺苷的水平有明显促进作用(P<0.05);固体培养基SCM的4种核苷类成分的水平较高.结论 培养基种类对蝉花培养物核苷类成分变化有直接的影响,确定蝉花虫草的生产工艺时,应充分考虑培养基的选择,同时结合产品成分和生产成本等因素进行综合评价.%Objective The HPLC method was established to simultaneously determine the contents of four kinds of nucleosides in culture of Cordyceps cicadae,and to analyze the effect of different culture medium on nucleoside component content.Methods The HPLC method was applied to determining the contents of four kinds of nucleosides such as uridine,guanosine,adenosine,and N6-(2-hydroxyethyl) adenosine in C.cicadae.Chromatography was carried out on a Waters Symmetry shield TMRP C18 column (250 mm ×4.6 mm,5 μ m) using

  1. EFFECT OF CULTURE MEDIUM ON BACTERIOCIN PRODUCTION BY LACTOBACILLUS RHAMNOSUS HN001 AND LACTOBACILLUS REUTERI ATCC 53608

    Directory of Open Access Journals (Sweden)

    Aguilar-Uscanga B. R.

    2013-06-01

    Full Text Available The aim of this study was to evaluate the effect of media on bacteriocin production by Lactobacillus rhamnosus HN001 and Lactobacillus reuteri ATCC 53608 using three different media: YPM, YPF and MRS supplemented with glucose and K2HPO4. The optimum temperature was 37°C and initial pH 6.5. Bacteriocin-like substances produced by tested bacteria in MRS medium supplemented with glucose and K2HPO4 exhibited a broad antimicrobial spectrum determined by well diffusion assay against indicator bacteria Listeria monocytogenes, Lactobacillus sakei, Enterococcus faecium, Lactobacillus delbrueckii, Lactobacillus acidophilus, but no antimicrobial spectrum against E. coli O157:H7, Salmonella typhimurium, Staphylococcus aureus, Bacillus cereus was detected. Bacteriocin was sensitive to protease IV, trypsin, pepsin and -amylases, but resistant to lipase. It was also resistant to detergents such as Tween 80, Triton-X and SDS. This bacteriocin was thermo-stable (resistant at 60°C, 90°C and 100°C for 30 min. Tested bacteria showed the best antimicrobial (bacteriocin-like activity after growth in MRS medium. Bacteriocin substances produced by tested bacteria showed promising thermo-stable technological properties.

  2. Optimization of the Culture Medium Composition to Improve the Production of Hyoscyamine in Elicited Datura stramonium L. Hairy Roots Using the Response Surface Methodology (RSM)

    Science.gov (United States)

    Ryad, Amdoun; Lakhdar, Khelifi; Majda, Khelifi-Slaoui; Samia, Amroune; Mark, Asch; Corinne, Assaf-Ducrocq; Eric, Gontier

    2010-01-01

    Traditionally, optimization in biological analyses has been carried out by monitoring the influence of one factor at a time; this technique is called one-variable-at-a-time. The disadvantage of this technique is that it does not include any interactive effects among the variables studied and requires a large number of experiments. Therefore, in recent years, the Response Surface Methodology (RSM) has become the most popular optimization method. It is an effective mathematical and statistical technique which has been widely used in optimization studies with minimal experimental trials where interactive factors may be involved. This present study follows on from our previous work, where RSM was used to optimize the B5 medium composition in [NO3−], [Ca2+] and sucrose to attain the best production of hyoscyamine (HS) from the hairy roots (HRs) of Datura stramonium elicited by Jasmonic Acid (JA). The present paper focuses on the use of the RSM in biological studies, such as plant material, to establish a predictive model with the planning of experiments, analysis of the model, diagnostics and adjustment for the accuracy of the model. With the RSM, only 20 experiments were necessary to determine optimal concentrations. The model could be employed to carry out interpolations and predict the response to elicitation. Applying this model, the optimization of the HS level was 212.7% for the elicited HRs of Datura stramonium, cultured in B5-OP medium (optimized), in comparison with elicited HRs cultured in B5 medium (control). The optimal concentrations, under experimental conditions, were determined to be: 79.1 mM [NO3−], 11.4 mM [Ca2+] and 42.9 mg/L of sucrose. PMID:21151467

  3. Optimization of the Culture Medium Composition to Improve the Production of Hyoscyamine in Elicited Datura stramonium L. Hairy Roots Using the Response Surface Methodology (RSM

    Directory of Open Access Journals (Sweden)

    Eric Gontier

    2010-11-01

    Full Text Available Traditionally, optimization in biological analyses has been carried out by monitoring the influence of one factor at a time; this technique is called one‑variable‑at‑a‑time. The disadvantage of this technique is that it does not include any interactive effects among the variables studied and requires a large number of experiments. Therefore, in recent years, the Response Surface Methodology (RSM has become the most popular optimization method. It is an effective mathematical and statistical technique which has been widely used in optimization studies with minimal experimental trials where interactive factors may be involved. This present study follows on from our previous work, where RSM was used to optimize the B5 medium composition in [NO3−], [Ca2+] and sucrose to attain the best production of hyoscyamine (HS from the hairy roots (HRs of Datura stramonium elicited by Jasmonic Acid (JA. The present paper focuses on the use of the RSM in biological studies, such as plant material, to establish a predictive model with the planning of experiments, analysis of the model, diagnostics and adjustment for the accuracy of the model. With the RSM, only 20 experiments were necessary to determine optimal concentrations. The model could be employed to carry out interpolations and predict the response to elicitation. Applying this model, the optimization of the HS level was 212.7% for the elicited HRs of Datura stramonium, cultured in B5-OP medium (optimized, in comparison with elicited HRs cultured in B5 medium (control. The optimal concentrations, under experimental conditions, were determined to be: 79.1 mM [NO3−], 11.4 mM [Ca2+] and 42.9 mg/L of sucrose.

  4. 中小型食品出口企业应用HACCP体系存在的问题及建议%Study on the Problems and Suggestions Existing HACCP System Implementation in Small and Medium Sized Export Food Enterprises

    Institute of Scientific and Technical Information of China (English)

    刘怀龙

    2015-01-01

    This paper analyzes the small and medium export food enterprises may encounter in the process of constructing the HACCP system, and discusses how to promote the small and medium export food enterprises to effectively apply food export HACCP system.%分析我国中小型食品出口企业构建HACCP体系存在的问题, 从多个角度探讨如何推动中小型食品出口企业有效应用HACCP体系.

  5. Salame elaborado com Lactobacillus plantarum fermentado em meio de cultura de plasma suíno Salami sausage prepared with Lactobacillus plantarum fermented in porcine plasma culture medium

    Directory of Open Access Journals (Sweden)

    Paulo Cezar Bastianello Campagnol

    2007-12-01

    Full Text Available Este trabalho teve por objetivo produzir uma cultura starter com uma cepa de Lactobacillus plantarum em um meio de cultura com plasma suíno e verificar a viabilidade de sua aplicação em salame. O meio de cultura foi preparado com plasma suíno e água destilada (1:1, pH 11,0. Após a esterilização, 300 mL foram adicionados de 400 mL de uma solução estéril de glicose e difosfato de potássio. A cepa de Lb. plantarum foi semeada no meio de cultura e submetida à fermentação em pH 7,0, durante 36 horas (100 rpm, 37 ± 0,1 °C. Ao alcançar a fase estacionária, a cultura foi centrifugada e ressuspendida em leite desnatado estéril, liofilizada e aplicada em salame. A influência do inóculo foi avaliada nas características microbiológicas, físico-químicas e sensoriais de salames. Os resultados encontrados foram comparados com tratamentos sem adição de cultura starter e com uma cultura comercial. O microrganismo Lb. plantarum teve um crescimento máximo de 9,82 Log UFC.mL-1, após 30 horas de fermentação. Os salames elaborados com a cultura starter produzida apresentaram uma queda de pH significativamente maior, e menor valor de atividade de água que os demais tratamentos. O microrganismo Lb. plantarum melhorou significativamente o sabor dos salames.The purpose of this work was to produce a starter culture with a strain of Lactobacillus plantarum in a porcine plasma culture medium and ascertain the viability of applying it in salami sausage. The culture medium was prepared with porcine plasma and distilled water (1:1, pH 11.0. After sterilization, 300 mL were added of 400 mL of a sterile solution of glucose and potassium diphosphate. The Lb. plantarum strain was inoculated into the culture medium and subjected to fermentation at pH 7.0 for 36 hours (100 rpm, 37 ± 0.1 °C. When the stationary phase was reached, the culture was centrifuged and resuspensed in sterile skimmed milk, lyophilized and applied to salami. An evaluation was

  6. Rapid Induction of Cerebral Organoids From Human Induced Pluripotent Stem Cells Using a Chemically Defined Hydrogel and Defined Cell Culture Medium.

    Science.gov (United States)

    Lindborg, Beth A; Brekke, John H; Vegoe, Amanda L; Ulrich, Connor B; Haider, Kerri T; Subramaniam, Sandhya; Venhuizen, Scott L; Eide, Cindy R; Orchard, Paul J; Chen, Weili; Wang, Qi; Pelaez, Francisco; Scott, Carolyn M; Kokkoli, Efrosini; Keirstead, Susan A; Dutton, James R; Tolar, Jakub; O'Brien, Timothy D

    2016-07-01

    Tissue organoids are a promising technology that may accelerate development of the societal and NIH mandate for precision medicine. Here we describe a robust and simple method for generating cerebral organoids (cOrgs) from human pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. By using no additional neural induction components, cOrgs appeared on the hydrogel surface within 10-14 days, and under static culture conditions, they attained sizes up to 3 mm in greatest dimension by day 28. Histologically, the organoids showed neural rosette and neural tube-like structures and evidence of early corticogenesis. Immunostaining and quantitative reverse-transcription polymerase chain reaction demonstrated protein and gene expression representative of forebrain, midbrain, and hindbrain development. Physiologic studies showed responses to glutamate and depolarization in many cells, consistent with neural behavior. The method of cerebral organoid generation described here facilitates access to this technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable. Tissue organoids are a promising technology with many potential applications, such as pharmaceutical screens and development of in vitro disease models, particularly for human polygenic conditions where animal models are insufficient. This work describes a robust and simple method for generating cerebral organoids from human induced pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. This method, by virtue of its simplicity and use of defined materials, greatly facilitates access to cerebral organoid technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable. ©AlphaMed Press.

  7. Sporulation of Clostridium cellulolyticum while grown in cellulose-batch and cellulose-fed continuous cultures on a mineral-salt based medium.

    Science.gov (United States)

    Desvaux, M; Petitdemange, H

    2002-03-01

    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.

  8. Strain and Culture Medium Optimization for Production Enhancement of Prodiginines from Marine-Derived Streptomyces sp.GQQ-10

    Institute of Scientific and Technical Information of China (English)

    LI Xueping; ZHANG Guojian; ZHU Tianjiao; LI Dehai; GU Qianqun

    2012-01-01

    A mutant(GQQ-M6)of a Sponge-Derived streptomyces sp.GQQ-10 obtained by UV-induced mutation was used for producing prodiginines(PGs).Single factor experiments and orthogonal array design(OAD)methods were employed for medium optimization.In the single factor method,the effects of soluble starch,glucose,soybean flour,yeast extract and sodium acetate on PGs production were investigated individually.In the subsequent OAD experiments,the concentrations of these 5 key nutritional components combined with salinity were further adjusted.The mutant strain GQQ-M6 gave a 2.2-fold higher PGs production than that of the parent strain;OAD experiments offered a PGs yield of 61mgL-1,which was 10 times higher than that of the initial GQQ-10 strain under the original cultivation mode.

  9. Strain and culture medium optimization for production enhancement of prodiginines from marine-derived Streptomyces sp. GQQ-10

    Science.gov (United States)

    Li, Xueping; Zhang, Guojian; Zhu, Tianjiao; Li, Dehai; Gu, Qianqun

    2012-09-01

    A mutant (GQQ-M6) of a Sponge-Derived streptomyces sp. GQQ-10 obtained by UV-induced mutation was used for producing prodiginines (PGs). Single factor experiments and orthogonal array design (OAD) methods were employed for medium optimization. In the single factor method, the effects of soluble starch, glucose, soybean flour, yeast extract and sodium acetate on PGs production were investigated individually. In the subsequent OAD experiments, the concentrations of these 5 key nutritional components combined with salinity were further adjusted. The mutant strain GQQ-M6 gave a 2.2-fold higher PGs production than that of the parent strain; OAD experiments offered a PGs yield of 61mg L-1, which was 10 times higher than that of the initial GQQ-10 strain under the original cultivation mode.

  10. Evaluation of the HB&L System for the Microbiological Screening of Storage Medium for Organ-Cultured Corneas

    Directory of Open Access Journals (Sweden)

    D. Camposampiero

    2013-01-01

    Full Text Available Aims. To compare HB&L and BACTEC systems for detecting the microorganisms contaminating the corneal storage liquid preserved at 31°C. Methods. Human donor corneas were stored at 4°C followed by preservation at 31°C. Samples of the storage medium were inoculated in BACTEC Peds Plus/F (aerobic microorganisms, BACTEC Plus Anaerobic/F (anaerobic microorganisms, and HB&L bottles. The tests were performed (a after six days of storage, (b end of storage, and (c after 24 hours of preservation in deturgescent liquid sequentially. 10,655 storage and deturgescent media samples were subjected to microbiological control using BACTEC (6-day incubation and HB&L (24-hour incubation systems simultaneously. BACTEC positive/negative refers to both/either aerobic and anaerobic positives/negatives, whereas HB&L can only detect the aerobic microbes, and therefore the positives/negatives depend on the presence/absence of aerobic microorganisms. Results. 147 (1.38% samples were identified positive with at least one of the two methods. 127 samples (134 identified microorganisms were positive with both HB&L and BACTEC. 14 HB&L+/BACTEC− and 6 BACTEC+/HB&L− were identified. Sensitivity (95.5%, specificity (99.8%, and positive (90.1% and negative predictive values (99.9% were high with HB&L considering a 3.5% annual contamination rate. Conclusion. HB&L is a rapid system for detecting microorganisms in corneal storage medium in addition to the existing methods.

  11. Evaluation of the HB&L System for the Microbiological Screening of Storage Medium for Organ-Cultured Corneas.

    Science.gov (United States)

    Camposampiero, D; Grandesso, S; Zanetti, E; Mazzucato, S; Solinas, M; Parekh, M; Frigo, A C; Gion, M; Ponzin, D

    2013-01-01

    Aims. To compare HB&L and BACTEC systems for detecting the microorganisms contaminating the corneal storage liquid preserved at 31°C. Methods. Human donor corneas were stored at 4°C followed by preservation at 31°C. Samples of the storage medium were inoculated in BACTEC Peds Plus/F (aerobic microorganisms), BACTEC Plus Anaerobic/F (anaerobic microorganisms), and HB&L bottles. The tests were performed (a) after six days of storage, (b) end of storage, and (c) after 24 hours of preservation in deturgescent liquid sequentially. 10,655 storage and deturgescent media samples were subjected to microbiological control using BACTEC (6-day incubation) and HB&L (24-hour incubation) systems simultaneously. BACTEC positive/negative refers to both/either aerobic and anaerobic positives/negatives, whereas HB&L can only detect the aerobic microbes, and therefore the positives/negatives depend on the presence/absence of aerobic microorganisms. Results. 147 (1.38%) samples were identified positive with at least one of the two methods. 127 samples (134 identified microorganisms) were positive with both HB&L and BACTEC. 14 HB&L+/BACTEC- and 6 BACTEC+/HB&L- were identified. Sensitivity (95.5%), specificity (99.8%), and positive (90.1%) and negative predictive values (99.9%) were high with HB&L considering a 3.5% annual contamination rate. Conclusion. HB&L is a rapid system for detecting microorganisms in corneal storage medium in addition to the existing methods.

  12. Application of a paper based device containing a new culture medium to detect Vibrio cholerae in water samples collected in Haiti.

    Science.gov (United States)

    Briquaire, Romain; Colwell, Rita R; Boncy, Jacques; Rossignol, Emmanuel; Dardy, Aline; Pandini, Isabelle; Villeval, François; Machuron, Jean-Louis; Huq, Anwar; Rashed, Shah; Vandevelde, Thierry; Rozand, Christine

    2017-02-01

    Cholera is now considered to be endemic in Haiti, often with increased incidence during rainy seasons. The challenge of cholera surveillance is exacerbated by the cost of sample collection and laboratory analysis. A diagnostic tool is needed that is low cost, easy-to-use, and able to detect and quantify Vibrio cholerae accurately in water samples within 18-24h, and perform reliably in remote settings lacking laboratory infrastructure and skilled staff. The two main objectives of this study were to develop and evaluate a new culture medium embedded in a new diagnostic tool (PAD for paper based analytical device) for detecting Vibrio cholerae from water samples collected in Haiti. The intent is to provide guidance for corrective action, such as chlorination, for water positive for V. cholerae epidemic strains. For detecting Vibrio cholerae, a new chromogenic medium was designed and evaluated as an alternative to thiosulfate citrate bile salts sucrose (TCBS) agar for testing raw water samples. Sensitivity and specificity of the medium were assessed using both raw and spiked water samples. The Vibrio cholerae chromogenic medium was proved to be highly selective against most of the cultivable bacteria in the water samples, without loss of sensitivity in detection of V. cholerae. Thus, reliability of this new culture medium for detection of V. cholerae in the presence of other Vibrio species in water samples offers a significant advantage. A new paper based device containing the new chromogenic medium previously evaluated was compared with reference methods for detecting V. cholerae from spiked water sample. The microbiological PAD specifications were evaluated in Haiti. More precisely, a total of 185 water samples were collected at five sites in Haiti, June 2014 and again in June 2015. With this new tool, three V. cholerae O1 and 17 V. cholerae non-O1/O139 strains were isolated. The presence of virulence-associated and regulatory genes, including ctxA, zot, ace, and tox

  13. Changes in the metabolic footprint of placental explant-conditioned medium cultured in different oxygen tensions from placentas of small for gestational age and normal pregnancies.

    LENUS (Irish Health Repository)

    Horgan, R P

    2012-01-31

    Being born small for gestational age (SGA) confers significantly increased risks of perinatal morbidity and mortality. Accumulating evidence suggests that an SGA fetus results from a poorly perfused and abnormally developed placenta. Some of the placental features seen in SGA, such as abnormal cell turnover and impaired nutrient transport, can be reproduced by culture of placental explants in hypoxic conditions. Metabolic footprinting offers a hypothesis-generating strategy to investigate factors absorbed by and released from this tissue in vitro. Previously, metabolic footprinting of the conditioned culture media has identified differences in placental explants cultured under normoxic and hypoxic conditions and between normal pregnancies and those complicated by pre-eclampsia. In this study we aimed to examine the differences in the metabolic footprint of placental villous explants cultured at different oxygen (O(2)) tensions between women who deliver an SGA baby (n = 9) and those from normal controls (n = 8). Placental villous explants from cases and controls were cultured for 96 h in 1% (hypoxic), 6% (normoxic) and 20% (hyperoxic) O(2). Metabolic footprints were analysed by Ultra Performance Liquid Chromatography coupled to an electrospray hybrid LTQ-Orbitrap Mass Spectrometry (UPLC-MS). 574 metabolite features showed significant difference between SGA and normal at one or more of the oxygen tensions. SGA explant media cultured under hypoxic conditions was observed, on a univariate level, to exhibit the same metabolic signature as controls cultured under normoxic conditions in 49% of the metabolites of interest, suggesting that SGA tissue is acclimatised to hypoxic conditions in vivo. No such behaviour was observed under hyperoxic culture conditions. Glycerophospholipid and tryptophan metabolism were highlighted as areas of particular interest.

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

    Science.gov (United States)

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

    2017-01-01

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

  15. Hypnosis, suggestion, and suggestibility: an integrative model.

    Science.gov (United States)

    Lynn, Steven Jay; Laurence, Jean-Roch; Kirsch, Irving

    2015-01-01

    This article elucidates an integrative model of hypnosis that integrates social, cultural, cognitive, and neurophysiological variables at play both in and out of hypnosis and considers their dynamic interaction as determinants of the multifaceted experience of hypnosis. The roles of these variables are examined in the induction and suggestion stages of hypnosis, including how they are related to the experience of involuntariness, one of the hallmarks of hypnosis. It is suggested that studies of the modification of hypnotic suggestibility; cognitive flexibility; response sets and expectancies; the default-mode network; and the search for the neurophysiological correlates of hypnosis, more broadly, in conjunction with research on social psychological variables, hold much promise to further understanding of hypnosis.

  16. Medium Optimization and Submerged Culture for Cenococcum geophilum Fr%土生空团菌培养基优化与摇瓶培养

    Institute of Scientific and Technical Information of China (English)

    薛丽宁; 曹凤娟; 赵慧英; 姚庆智; 闫伟

    2013-01-01

    Based on the results of the single-factor tests, the medium components of four strains of Cg (CgSO1, CgSB2, CgO5 and SPOP2) were optimized by the orthogonal tests. The results showed that the four strains of Cg required different medium components and different carbon sources, nitrogen sources, microelements, mineral salt components and vitamins. The best carbon source and nitrogen source for CgSOl, CgSB2 and CgO5 were glucose + wort and beef extract while SPOP2 were wort and ammonium tartrate. The appropriate amount of microelements, mineral salts and Vitamin Bl enhanced the growth of CgSOl, CgSB2, CgO5 and SPOP2. The results of the optimized medium by plate culture could be used in liquid culture and the biomass of the four strains in liquid culture were 13.2, 12.4, 18.9 and 15.1 g/L, respectively, which all met the requirement of liquid fermentation production. Fig 1, Tab 6, Ref 17%在单因素试验的基础上,通过正交试验对CgSO1、CgSB2、CgO5和SPOP2这4株土生空团菌[Cenococcum geophilum Fr.(Cg)]的培养基配方分别进行了优化.结果发现,来源不同的Cg菌株对培养基组分需求存在一定差异,碳氮源、微量元素、无机盐以及维生素都不相同,CgSO1、CgSB2、CgO5对葡萄糖和麦芽汁的混合C源以及牛肉浸膏利用最好,SPOP2对麦芽汁以及酒石酸铵的利用最好,适量的微量元素、无机盐以及维生素B1会促进Cg的生长.经平板培养基优化后的培养基配方可应用到液体培养中,这4株Cg菌经液体培养后菌丝产量分别达到了13.2、12.4、18.9和15.1g/L,均可满足液体发酵的生产要求.

  17. Medium Optimization for Potato Plantlet Culture%马铃薯试管苗快繁培养基的优化研究

    Institute of Scientific and Technical Information of China (English)

    刘海英

    2014-01-01

    The effect of mineral salt concentrationson potato plantlet growth were studied to solve the problems about the long reproductive cycle and low survival rate for the potato plantlet culture.Adopted multi-factor orthogonal experiment L9 (34) design was used to change the MS medium concentration of CaCl2,Ca(NO3)2,KH2PO4,KNO3 to screen the optimum concentration for potato tissue culture medium preparation.The results showed that CaCl2 0mmol/L,Ca(NO3) 2 20mmol/L,KH2PO4 6mmol/L,KNO3 18mmol/L was the best combination for the potato plantlet culture medium.%针对马铃薯试管苗快繁继代繁殖周期长和移栽后成活率低的实际问题,以马铃薯品种克新l号脱毒试管苗为试验材料,采用多因子正交试验L9(34)设计方法,以MS培养基中4种无机盐CaCl2 、Ca(NO3)2、KH2 PO4、KNO3为试验因素,研究不同无机盐浓度对马铃薯试管苗生长的影响.结果表明,在CaCl2浓度为0mmol/L、Ca(NO3)2浓度为20mmol/L、KH2PO4浓度为6mmol/L、KNO3浓度为18 mmol/L条件下,克新1号马铃薯试管苗的株高、茎粗、根系、茎叶鲜重生长达到最好.本研究结果可为马铃薯组培快繁过程中培养基制备提供依据.

  18. Study of the interaction of inorganic and organic compounds of cell culture medium with a Ti surface.

    Science.gov (United States)

    Burgos-Asperilla, L; García-Alonso, M C; Escudero, M L; Alonso, C

    2010-02-01

    The interaction between Ti and each component of Dulbecco's modified Eagle's medium was studied in depth using different techniques, such as the measurement of the corrosion potential, electrochemical impedance spectroscopy and polarization curves. The characterization of metal surfaces was carried out by scanning electron microscopy and X-ray photoelectron spectroscopy (XPS). The adsorption process of each component was studied using the quartz crystal balance (QCM). The QCM and XPS results reveal that the adsorption kinetics for phosphate and calcium ions is slow. However, the bovine serum albumin (BSA) totally covers the Ti surface rapidly. Because the passive film (titanium oxide) has acidic hydroxyl groups, the calcium ions would have a bridging effect on the electrostatic adsorption of phosphate ions as well as that of BSA. The polarization curves reveal that the adsorbed glucose permits the ionic diffusion of the oxygen to the electrode, while the BSA and fetal bovine serum (FBS) adsorbed after 7 days of immersion act as a diffusive barrier. The impedance measurement and data fitting to the electrical equivalent circuit model show that the resistance of the proteins/TiO(2) interface, for Ti immersed in FBS, is higher than those obtained for BSA, due to the proteins present in the solution as well as the fact that the adsorbed proteins on the surface are greater.

  19. Size distribution of fullerenol nanoparticles in cell culture medium and their influence on antioxidative enzymes in Chinese hamster ovary cells

    Directory of Open Access Journals (Sweden)

    Srđenović Branislava U.

    2015-01-01

    Full Text Available Fullerenol (C60(OH24 nanoparticles (FNP have a significant role in biomedical research due to their numerous biological activities, some of which are cytoprotective and antioxidative properties. The aim of this study was to measure distribution of fullerenol nanoparticles and zeta potential in cell medium RPMI 1640 with 10% fetal bovine serum (FBS and to investigate the influence of FNP on Chinese hamster ovary cells (CHO-K1 survival, as well as to determine the activity of three antioxidative enzymes: superoxide-dismutase, glutathione-reductase and glutathione-S-transferase in mitomycin C-treated cell line. Our investigation implies that FNP, as a strong antioxidant, influence the cellular redox state and enzyme activities and thus may reduce cell proliferation, which confirms that FNP could be exploited for its use as a cytoprotective agent.[Projekat Ministarstva nauke Republike Srbije, br. III45005 i Pokrajinski Sekretarijat za nauku i tehnološki razvoj Vojvodine, grant number 114-451-2056/2011-01

  20. Chemically Defined Medium and Caenorhabditis elegans: A Powerful Approach

    Science.gov (United States)

    Szewczyk, N. J.; Kozak, E.; Conley, C. A.

    2003-01-01

    C. elegans has been established as a powerful genetic system. Growth in a chemically defined medium (C. elegans Maintenance Medium (CeMM)) now allows standardization and systematic manipulation of the nutrients that animals receive. Liquid cultivation allows automated culturing and experimentation and should be of me in large-scale growth and screening of animals. Here we present our initial results from developing culture systems with CeMM. We find that CeMM is versatile and culturing is simple. CeMM can be used in a solid or liquid state, it can be stored unused for at least a year, unattended actively growing cultures may be maintained longer than with standard techniques, and standard C. elegans protocols work well with animals grown in defined medium. We also find that there are caveats of using defined medium. Animals in defined medium grow more slowly than on standard medium, appear to display adaptation to the defined medium, and display altered growth rates as they change defined medium composition. As was suggested with the introduction of C. elegans as a potential genetic system, use of defined medium with C. elegans should prove a powerful tool.

  1. Thinking and suggestions on the management of the medium term financial planning in the meteorological department%气象部门推进中期财政规划管理的思考与建议

    Institute of Scientific and Technical Information of China (English)

    林霖; 李博

    2016-01-01

    根据《国务院关于实行中期财政规划管理的意见》与《财政部关于推进中央部门中期财政规划管理的意见》,气象部门积极推进中期财政规划管理工作。当前的中期财政规划管理,既有项目库储备少、资金规模小,基层预算编制时间紧、缺少科学编制依据,重工程建设、轻运行维护等现实问题,也要考虑中期财政规划和气象发展五年规划的匹配、年度平衡和跨期平衡的衔接、全口径预算基础的统筹与宏观指导的长远问题。在此基础上,气象部门推进中期财政规划管理需瞄准长远,做好基础。顶层设计要与实际需求的测算相结合,夯实中期财政规划管理工作的基础,合理制定战略决策、中长期规划及绩效目标。%According to the State Council on the implementation of opinions "and" the Ministry of Finance on promoting medium-term fiscal planning and management of the central departments of medium-term fiscal planning and management opinions, meteorological departments to actively promote the work of the management of financial planning. The medium-term medium-term fiscal plan management at present, both the project library reserves less, small scale capital budget, time is tight the lack of scientific basis, and heavy construction, light maintenance and other practical problems, but also to consider, medium-term fiscal planning and meteorological development five year plan, the annual balance and intertemporal balance convergence, the problem of long-term overall budget based full caliber and macro guidance. On this basis, the meteorological departments to promote medium-term fiscal be aimed at long-term planning and management, make base for the top-level design to calculate with the actual needs of the combination, laying a solid foundation for medium-term fiscal planning and management work, make reasonable strategic decision, Medium and long term planning and performance

  2. [Influence of genotype, explant type and component of culture medium on in vitro callus induction and shoot organogenesis of tomato (Solanum lycopersicum L.)].

    Science.gov (United States)

    Khaliluev, M R; Bogoutdinova, L R; Baranova, G B; Baranova, E N; Kharchenko, P N; Dolgov, S V

    2014-01-01

    The influence of explant type as well as of the type of growth regulators and concentration on callus induction processes and somatic organogenesis of shoots was studied in vitro on four tomato genotypes of Russian breeding. Cytological study of callus tissue was conducted. It was established that tomato varieties possess a substantially greater ability to indirect shoot organogenesis compared with the F1 hybrid. The highest frequency of somatic organogenesis of shoots, as well as their number per explant, was observed for most of the genotypes studied during the cultivation of cotyledons on Murashige-Skoog culture medium containing 2 mg/l of zeatin in combination with 0.1 mg/l of 3-indoleacetic acid. An effective protocol of indirect somatic organogenesis of shoots from different explants of tomato varieties with a frequency of more than 80% was developed.

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

    Science.gov (United States)

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

    2016-08-01

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

  4. Efficacy of in vitro embryo transfer in lactating dairy cows using fresh or vitrified embryos produced in a novel embryo culture medium.

    Science.gov (United States)

    Block, J; Bonilla, L; Hansen, P J

    2010-11-01

    Objectives were to determine whether pregnancy success could be improved in lactating cows with timed embryo transfer when embryos were produced in vitro using a medium designed to enhance embryo development and survival after cryopreservation. In experiment 1, embryos (n=569 to 922) were cultured in either modified synthetic oviduct fluid or a serum-free medium, Block-Bonilla-Hansen-7 (BBH7). Development to the blastocyst stage was recorded at d 7, and selected blastocysts (n=79 to 114) were vitrified using open pulled straws. Culture of embryos in BBH7 increased development to the blastocyst stage (41.9±2.0 vs. 14.7±2.0%) and advanced blastocyst stages (expanded, hatching, hatched; 31.1±1.3 vs. 6.4±1.3%) at d 7 and resulted in higher hatching rates at 24h postwarming compared with embryos cultured in modified synthetic oviduct fluid (59.0±0.5 vs. 26.7±0.5%). In experiment 2, embryos were produced using X-sorted semen and cultured in BBH7. At d 7 after insemination, embryos were transferred fresh or following vitrification. Lactating Holstein cows were either subjected to timed artificial insemination (TAI) on the day of presumptive ovulation or used as embryo recipients 7 d later. Embryo recipients received an embryo if a corpus luteum was present. The percentage of cows pregnant at d 32, 46, and 76 of gestation was higher among cows that received fresh embryos compared with TAI cows or cows that received vitrified embryos. At d 76, for example, the proportion and percentage pregnant was 47/150 (31.3%) for cows subjected to TAI, 48/95 (50.5%) for cows receiving fresh embryos, and 39/141 (27.7%) for cows receiving a vitrified embryo. No difference was observed in the percentage of cows pregnant among TAI cows and those that received vitrified embryos. There was a service or transfer number × treatment interaction because differences in pregnancy rate between embryo transfer recipients and cows bred by TAI were greater for cows with more than 3 services or

  5. Enrichment of prostate cancer stem-like cells from human prostate cancer cell lines by culture in serum-free medium and chemoradiotherapy.

    Science.gov (United States)

    Wang, Lei; Huang, Xing; Zheng, Xinmin; Wang, Xinghuan; Li, Shiwen; Zhang, Lin; Yang, Zhonghua; Xia, Zhiping

    2013-01-01

    The discovery of rare subpopulations of cancer stem cells (CSCs) has created a new focus in cancer research. As CSCs demonstrate resistance to chemoradiation therapy relative to other cancer cells, this allows the enrichment of CSC populations by killing apoptosis-susceptible cancer cells. In this study, three commonly used human prostate cancer (PCa) cell lines (DU145, PC-3 and LNCaP) were examined for their expression of the putative stem cell markers CD133 and CD44 via flow cytometric analysis. Under normal culture conditions, CD133(+)/CD44(+) cells were only present in the DU145 cell line, and comprised only a minor percentage (0.1% ± 0.01%) of the total population. However, the proportion of these CD133(+)/CD44(+) prostate CSCs could be increased in these cell lines via culture in serum-free medium (SFM), or through chemotherapy or radiotherapy. Indeed, after culture in SFM, the proportion of CD133(+)/CD44(+) cells in DU145 and PC-3 had increased to 10.3% and 3.0%, respectively. Moreover, the proportion had increased to 9.8% enriched by chemotherapy and 3.5% by radiotherapy in DU145. Colony-formation tests, cell invasion assays, and tumor xenografts in BALB/c nude mice were used to evaluate the stem cell properties of CD133(+)/CD44(+) PCa cells that were isolated via fluorescence-activated cell sorting (FACS). CD133(+)/CD44(+) cells had an enhanced colony-formation capability and invasive ability in vitro, and displayed greater tumorigenic properties in vivo. These results demonstrate the presence of CD133(+)/CD44(+) prostate CSCs in established PCa cell lines and that populations of these cells can be enriched by culture in SFM or chemoradiotherapy. Finding novel therapies to override chemoradiation resistance in the prostate CSCs is the key to improve long-term results in PCa management.

  6. In vitro culture of early secondary preantral follicles in hanging drop of ovarian cell-conditioned medium to obtain MII oocytes from outbred deer mice.

    Science.gov (United States)

    Choi, Jung Kyu; Agarwal, Pranay; He, Xiaoming

    2013-12-01

    The ovarian follicle (each contains a single oocyte) is the fundamental functional tissue unit of mammalian ovaries. In humans, it has been long held true that females are born with a maximum number of follicles (or oocytes) that are not only nonrenewable, but also undergoing degeneration with time with a sharply decreased oocyte quality after the age of ∼35. Therefore, it is of importance to isolate and bank ovarian follicles for in vitro culture to obtain fertilizable oocytes later, to preserve the fertility of professional women who may want to delay childbearing, young and unmarried women who may lose gonadal function because of exposure to environmental/occupational hazards or aggressive medical treatments, such as radiation and chemotherapy, and even endangered species and breeds. Although they contributed significantly to the understanding of follicle science and biology, most studies reported to date on this topic were done using the man-made, unnatural inbred animal species. It was found in this study that the conventional two-dimensional microliter drop and three-dimensional hanging drop (HD) methods, reported to be effective for in vitro culture of preantral follicles from inbred mice, are not directly transferrable to outbred deer mice. Therefore, a modified HD method was developed in this study to achieve a much higher (>5 times compared to the best conventional methods) percentage of developing early secondary preantral follicles from the outbred mice to the antral stage, for which, the use of an ovarian cell-conditioned medium and multiple follicles per HD were identified to be crucial. It was further found that the method for in vitro maturation of oocytes in antral follicles obtained by in vitro culture of preantral follicles could be very different from that for oocytes in antral follicles obtained by hormone stimulation in vivo. Therefore, this study should provide important guidance for establishing effective protocols of in vitro follicle

  7. Effect of medium variations (zinc supplementation during oocyte maturation, perifertilization pH, and embryo culture protein source) on equine embryo development after intracytoplasmic sperm injection.

    Science.gov (United States)

    Choi, Young-Ho; Gibbons, John R; Canesin, Heloísa S; Hinrichs, Katrin

    2016-10-15

    Prospective studies were conducted to help define procedural factors affecting in vitro embryo production via intracytoplasmic sperm injection (ICSI) of equine oocytes. In experiment 1, use of 10% fetal bovine serum as a protein source in embryo culture medium resulted in a higher blastocyst rate than did use of a combination of 3% fetal bovine serum, 3% equine preovulatory follicular fluid, and 4% human serum substitute (37% vs. 15%, respectively, P zinc supplementation (0, 0.5, 1, or 1.5 μg/mL) during IVM was examined. There were no significant differences in rates of cleavage or blastocyst development (20%-31%). However, the proportion of blastocysts that developed on Day 7 for the added-zinc treatments was significantly higher than that for the control treatment (45% vs. 8%). In experiment 3, we tested whether use of high-pH medium (pH 8.0-8.4) during ICSI procedures would improve blastocyst rate when sperm with low cleavage rates after ICSI was used. When high-pH conditions were used for sperm preparation and also for the first 2 hours of incubation of injected oocytes after ICSI, the cleavage rate was unaffected but no blastocysts developed (0% vs. 24% for control). When high-pH conditions were used for sperm preparation only, the blastocyst rate was 37%. This was repeated using sperm from a second stallion; there was no significant difference in cleavage or blastocyst rates between sperm preparation in high pH vs. control medium. These findings add to our knowledge of factors affecting in vitro production of equine embryos.

  8. Study on Culture Medium of Ganoderma lucidum in Zhuhai%珠海灵芝栽培基质配方的筛选研究

    Institute of Scientific and Technical Information of China (English)

    张长海; 沈汉国; 陈继敏; 潘丽晶; 张妙彬; 黄聪灵; 林淑芬

    2013-01-01

      为筛选出适合珠海地区灵芝栽培的基质配方,采用不同木屑原料、不同配比等进行灵芝优良菌株的栽培研究。研究结果发现,以台湾相思为木屑原料以及木屑与棉籽壳的比例为1:1时,灵芝的菌丝生长、子实体形态特征、产量和有效成分等4项综合性状最优。综合来看,珠海本地的灵芝高产栽培基质配方为:木屑35%,棉籽壳35%,麦麸20%,玉米粉6%,糖1%,石膏1%,碳酸钙1%,磷酸二氢钾1%。此配方含水量60%。%In order to screen the suitable culture medium of Ganoderma lucidum in Zhuhai, four kinds of wood materials and different ratio of wood materials for Ganoderma lucidum cultivation were studied in this paper. The results showed that when cultivation with Taiwan Acacia sawdust and the ratio of wood material: cotton seed shell being 1:1, the comprehensive characters including mycelium growth, fruiting body morphology, yield and active components of Ganoderma lucidum were the best. In general, the optimized culture medium formula for Ganoderma lucidum cultivation in Zhuhai was: wood 35%, cottonseed shell 35%, wheat bran 20%, corn powder 6%, sugar 1%, gypsum 1%, calcium carbonate 1%, potassium dihydrogen phosphate 1%, water 60%.

  9. Attitudes to Suggestions

    Institute of Scientific and Technical Information of China (English)

    PETER; JOHNSON

    2007-01-01

    As an Australian expat teaching English in China for over four years, I often encourage my students to not only learn the English language but also try to understand Western culture. This includes the fact that Westerners frequently initiate proactive suggestions on any aspects of soci-

  10. Optimization of culture medium for anaerobic production of rhamnolipid by recombinant Pseudomonas stutzeri Rhl for microbial enhanced oil recovery.

    Science.gov (United States)

    Zhao, F; Mandlaa, M; Hao, J; Liang, X; Shi, R; Han, S; Zhang, Y

    2014-08-01

    Response surface methodology was employed to enhance the anaerobic production of rhamnolipid by recombinant Pseudomonas stutzeri Rhl. Glycerol is a promising carbon source used to anaerobically produce rhamnolipid. In a Plackett-Burman design, glycerol, KH2 PO4 and yeast extract were significant factors. The proposed optimized medium contained the following: 46·55 g l(-1) glycerol; 3 g l(-1) NaNO3 ; 5·25 g l(-1) K2 HPO4 ·3H2 O; 5·71 g l(-1) KH2 PO4 ; 0·40 g l(-1) MgSO4 ·7H2 O; 0·13 g l(-1) CaCl2 ; 1·0 g l(-1) KCl; 1·0 g l(-1) NaCl; and 2·69 g l(-1) yeast extract. Using this optimized medium, we obtained an anaerobic yield of rhamnolipid of 3·12 ± 0·11 g l(-1) with a 0·85-fold increase. Core flooding test results also revealed that Ps. stutzeri Rhl grown in an optimized medium enhanced the oil recovery efficiency by 15·7%, which was 6·6% higher than in the initial medium. Results suggested that the optimized medium is a promising nutrient source that could effectively mobilize oil by enhancing the in situ production of rhamnolipid. The ex situ application of rhamnolipid for microbial enhanced oil recovery (MEOR) is costly and complex in terms of rhamnolipid production, purification and transportation. Compared with ex situ applications, the in situ production of rhamnolipid in anaerobic oil reservoir is more advantageous for MEOR. This study is the first to report the anaerobic production optimization of rhamnolipid. Results showed that the optimized medium enhanced not only the anaerobic production of rhamnolipid but also crude oil recovery. © 2014 The Society for Applied Microbiology.

  11. The application of phytone in the pneumococcal culture medium%植物蛋白胨在肺炎链球菌培养基中的应用

    Institute of Scientific and Technical Information of China (English)

    邓雪莲; 颜熙熙; 张锐; 王静; 毕俊虹

    2014-01-01

    目的 用植物蛋白胨替代目前肺炎链球菌(pneumococcus,Pn)培养基中的动物蛋白胨,以保证生物制品的安全性.方法 比较大豆蛋白胨与胰蛋白胨的各种成分含量.分别以大豆蛋白胨和胰蛋白胨制备培养基进行9个型Pn大罐培养,比较两种培养基培养对Pn生长和粗制多糖含量的影响.结果 大豆蛋白胨的总氮含量、氨氮含量和消化系数与胰蛋白胨相近,但大豆蛋白胨的总碳水化合物含量(336.20 μg/g)明显高于胰蛋白胨的总碳水化合物含量(10.56 μg/g).植物源性培养基培养获得的各型Pn浓度(t=3.851,P<0.05)和粗制多糖含量(t=3.853,P<0.05)明显高于动物源性培养基培养.结论 植物蛋白胨可替代动物蛋白胨来制备Pn培养基.%Objective To replace animal peptone by phytone in the pneumococcal culture medium in order to ensure the safety of biological products.Methods Contents of various components in soy peptone were compared with those in animal peptone.Two kinds of pneumococcal culture media were prepared by using soy peptone and tryptone respectively to culture 9 types of pneumococcus in the large fermentation tank.The influence of 2 kinds of pneumococcal culture media on pneumococcal growth and content of crude polysaccharide were compared.Results The total nitrogen content,ammonia-nitrogen content and digestion coefficient of soy peptone were similar to tryptone,but the total carbohydrate content of soy peptone (336.20 μg/g) was significantly higher than that of tryptone (10.56 μg/g).The concentrations (t =3.851,P < 0.05) and the contents of crude polysaccharide (t =3.853,P < 0.05) of different types of pneumococcus harvested from the plant-derived culture media were significantly higher than those harvested from the animalderived culture media.Conclusion Animal peptone can be substituted by phytone to prepare the pneumococcal culture medium.

  12. A Clostridium Group IV Species Dominates and Suppresses a Mixed Culture Fermentation by Tolerance to Medium Chain Fatty Acids Products

    Science.gov (United States)

    Andersen, Stephen J.; De Groof, Vicky; Khor, Way Cern; Roume, Hugo; Props, Ruben; Coma, Marta; Rabaey, Korneel

    2017-01-01

    A microbial community is engaged in a complex economy of cooperation and competition for carbon and energy. In engineered systems such as anaerobic digestion and fermentation, these relationships are exploited for conversion of a broad range of substrates into products, such as biogas, ethanol, and carboxylic acids. Medium chain fatty acids (MCFAs), for example, hexanoic acid, are valuable, energy dense microbial fermentation products, however, MCFA tend to exhibit microbial toxicity to a broad range of microorganisms at low concentrations. Here, we operated continuous mixed population MCFA fermentations on biorefinery thin stillage to investigate the community response associated with the production and toxicity of MCFA. In this study, an uncultured species from the Clostridium group IV (related to Clostridium sp. BS-1) became enriched in two independent reactors that produced hexanoic acid (up to 8.1 g L−1), octanoic acid (up to 3.2 g L−1), and trace concentrations of decanoic acid. Decanoic acid is reported here for the first time as a possible product of a Clostridium group IV species. Other significant species in the community, Lactobacillus spp. and Acetobacterium sp., generate intermediates in MCFA production, and their collapse in relative abundance resulted in an overall production decrease. A strong correlation was present between the community composition and both the hexanoic acid concentration (p = 0.026) and total volatile fatty acid concentration (p = 0.003). MCFA suppressed species related to Clostridium sp. CPB-6 and Lactobacillus spp. to a greater extent than others. The proportion of the species related to Clostridium sp. BS-1 over Clostridium sp. CPB-6 had a strong correlation with the concentration of octanoic acid (p = 0.003). The dominance of this species and the increase in MCFA resulted in an overall toxic effect on the mixed community, most significantly on the Lactobacillus spp., which resulted in a decrease in total

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

    Directory of Open Access Journals (Sweden)

    Watson Andrew J

    2007-01-01

    Full Text Available Abstract Background Mechanisms that confer an ability to respond positively to environmental osmolarity are fundamental to ensuring embryo survival during the preimplantation period. Activation of p38 mitogen-activated protein kinase (MAPK occurs following exposure to hyperosmotic treatment. Recently, a novel scaffolding protein called Osmosensing Scaffold for MEKK3 (OSM was linked to p38 MAPK activation in response to sorbitol-induced hypertonicity. The human ortholog of OSM is cerebral cavernous malformation 2 (CCM2. The present study was conducted to investigate whether CCM2 is expressed during mouse preimplantation development and to determine whether this scaffolding protein is associated with p38 MAPK activation following exposure of preimplantation embryos to hyperosmotic environments. Results Our results indicate that Ccm2 along with upstream p38 MAPK pathway constituents (Map3k3, Map2k3, Map2k6, and Map2k4 are expressed throughout mouse preimplantation development. CCM2, MAP3K3 and the phosphorylated forms of MAP2K3/MAP2K6 and MAP2K4 were also detected throughout preimplantation development. Embryo culture in hyperosmotic media increased p38 MAPK activity in conjunction with elevated CCM2 levels. Conclusion These results define the expression of upstream activators of p38 MAPK during preimplantation development and indicate that embryo responses to hyperosmotic environments include elevation of CCM2 and activation of p38 MAPK.

  14. Structural investigation and response surface optimisation for improvement of kefiran production yield from a low-cost culture medium.

    Science.gov (United States)

    Ghasemlou, Mehran; Khodaiyan, Faramarz; Jahanbin, Kambiz; Gharibzahedi, Seyed Mohammad Taghi; Taheri, Salman

    2012-07-15

    Kefiran, a water-soluble heteropolysaccharide with molecular weight of 1.35×10(6)Da and a specific optical rotation of +64° (c 1.0, H2O), was isolated from kefir grains grown in cheese whey and further purified through DEAE-Sepharose XK26. Response surface methodology was employed to optimise the culture conditions for kefiran production from kefir grains to be lactose concentration 67 g/l, yeast extract 13g/l, pH 5.7 and temperature 24°C. Intrinsic viscosity was 5.84 dl/g using the Huggins extrapolation and 5.53 dl/g using the Kramer extrapolation. Monosaccharide analysis revealed that kefiran is composed of glucose (Glc) and galactose (Gal) in a relative molar ratio of 1.0:1.1. Its structural features were elucidated by a combination of FT-IR, methylation and GC-MS analysis, periodate oxidation-Smith degradation, partial acid hydrolysis and NMR spectroscopy ((1)H, (13)C and HMBC). The data obtained indicated that kefiran possessed a backbone of (1→6)-linked Glc, (1→3)-linked Gal, (1→4)-linked Gal, (1→4)-linked Glc and (1→2,6)-linked Gal, with a branch attached to O-2 of Gal residues and terminated with Glc residues.

  15. Scopoletin uptake from culture medium and accumulation in the vacuoles after conversion to scopolin in 2,4-D-treated tobacco cells.

    Science.gov (United States)

    Taguchi; Fujikawa; Yazawa; Kodaira; Hayashida; Shimosaka; Okazaki

    2000-02-21

    Tobacco (Nicotiana tabacum L. Bright Yellow) T-13 cell line has the ability to produce scopoletin endogenously and release some of it into the culture medium. We investigated the mechanism of scopoletin uptake following treatment of a tobacco culture with 2,4-dichlorophenoxyacetic acid (2,4-D). Addition of [14C]-labeled scopoletin showed that scopoletin was taken up by 2,4-D-treated cells and converted to scopolin, a 7-O-glucoside of scopoletin. This uptake of scopoletin began 6 h after 2,4-D addition to the cells. Experiments using several inhibitors showed that this uptake was energy-dependent. The phenomenon of 2,4-D-stimulated uptake was observed only for 7-hydroxycoumarins, such as scopoletin, umbelliferone and esculetin. To further investigate the site for scopoletin accumulation, we separated the vacuoles from T-13 cells and quantified the coumarin contents in this fraction. Most of the scopoletin in the vacuoles was present as glucoconjugate, scopolin. Moreover, glucosylation activity was absent from isolated vacuoles and, therefore, is likely to be located in the cytosol. Therefore, we can state that 2,4-D treatment of tobacco cells stimulated scopoletin uptake. The scopoletin was converted into scopolin in the cytoplasm, and then transferred into the vacuoles.

  16. Production of Gymnemic Acid Depends on Medium, Explants, PGRs, Color Lights, Temperature, Photoperiod, and Sucrose Sources in Batch Culture of Gymnema sylvestre

    Directory of Open Access Journals (Sweden)

    A. Bakrudeen Ali Ahmed

    2012-01-01

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

  17. Adding of ascorbic acid to the culture medium influences the antioxidant status and some biochemical parameters in the hen granulosa cells.

    Science.gov (United States)

    Capcarova, M; Kolesarova, A; Kalafova, A; Bulla, J; Sirotkin, A V

    2015-07-01

    The aim of the present study was to determine the activity of superoxide dismutase (SOD), total antioxidant status (TAS) of the hen granulosa cells, and selected biochemical parameters, including calcium, phosphorus, sodium, potassium, glucose, cholesterol, proteins, in the culture medium of granulosa cells after exposing them to ascorbic acid in vitro conditions. Ovarian granulosa cells of hens were incubated with various doses of ascorbic acid (E1 0.09 mg/ml, E2 0.13 mg/ml, E3 0.17 mg/ml, E4 0.33 mg/ml, E5 0.5 mg/ml). Ascorbic acid did not manifest antioxidant potential and higher doses of ascorbic acid (0.17; 0.33 and 0.5 mg/ml) decreased the activity of SOD in granulosa cells. Vitamin application resulted in a significantly (p<0.05) higher accumulation of Na+ and K+ in culture media of granulosa cells and decreased the concentration of glucose and proteins. These results indicate that ascorbic acid might be involved in the regulation of selected biochemical and physiological processes in ovarian granulosa cells.

  18. Production of gymnemic acid depends on medium, explants, PGRs, color lights, temperature, photoperiod, and sucrose sources in batch culture of Gymnema sylvestre.

    Science.gov (United States)

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

    2012-01-01

    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.

  19. Spore germination, early development and some notes on the effects of in vitro culture medium on Frullania ericoides (Nees Mont. (Frullaniaceae, Marchantiophyta

    Directory of Open Access Journals (Sweden)

    Juliana da Costa Silva-e-Costa

    Full Text Available ABSTRACT In bryophytes, establishment can occur by a sexual or asexual process, but the production of spores enables colonization of a wider range of habitats and substrates than can asexual propagules. Successful germination is critical for establishment in a new environment. This paper addresses germination and sporeling development in Frullania ericoides, a leafy liverwort species. Fresh spores were inoculated in vitro in different culture strengths of Knop’s nutrient solution (one-fourth strength, half strength, full strength, one and a half strength and double strength, in order to evaluate the effects of this solution on spore germination and on the development of external protonema. On the first assessment, spore germination was observed at all the concentrations. Germination was endosporic, with cell division and proliferation, resulting in a globular protonema, within the spore wall. Beginning at the fourth week, the development of tightly concave primordial leaves was observed in all but the double-strength medium. Throughout the period of study, the treatments with lower concentrations exhibited external protonema with greater lengths. The double-strength treatment was statistically different from other treatments in at least two parameters. The results of this study demonstrate the potential of in vitro culture techniques for bryophyte spore studies and germplasm preservation.

  20. community Participation for culturing Local Strain of bacillus Thuringiensis H-14 in coconut Medium to controlanopheles Sundaicus Larvae at kampung Laut, cilacap regency

    Directory of Open Access Journals (Sweden)

    Wiwik Trapsilowati

    2015-01-01

    Full Text Available background: Community participation was necessary for culturinglocal strain of Bacillus thuringiensis H-14 in coconut medium and adoption this bioinsecticide to control Anopheles sundaicus. This study aimed to evaluate community participation for culturing local strain of B. thuringiensis H-14 using coconut media and spreading the results of inoculation against Anopheles sundaicus larvae. Methods: The study design was a quasi-experiment. Reseach was conducted in February-November 2007 in Kampung Laut, Cilacap. Samples were 30 person which were choosed purposively. The sample criterias were at least 15 years of age and have a pond. Community participation was evaluated for 6 times using checklist. results: Results showed that all respondents (> 80% could have on every element of 9 elements properly. Ninety percent of respondents could prepare coconut independently and made local strains of B. thuringiensis H-14 inoculation in coconuts properly (96. 67%. The density of An. sundaicuslarvae was decreased by 80–100% after 1 day spreading with culture of local strain of B. thuringiensis H-14. conclusion: Communities could participate independently for culturinglocal strain of B. thuringiensis H-14 in coconuts to control An. sundaicus larvae. The inoculation capable todecrease An. sundaicus larvae density 80–100% after 1 day. Further research was recomended to calculate cost benefit and cost effectiveness.

  1. Brewer's spent grain and corn steep liquor as alternative culture medium substrates for proteinase production by Streptomyces malaysiensis AMT-3

    Directory of Open Access Journals (Sweden)

    Rodrigo Pires do Nascimento

    2011-12-01

    Full Text Available Brewer's spent grain and corn steep liquor or yeast extract were used as the sole organic forms for proteinase production by Streptomyces malaysiensis in submerged fermentation. The influence of the C and N concentrations, as well as the incubation periods, were assessed. Eight proteolytic bands were detected through gelatin-gel-electrophoresis in the various extracts obtained from the different media and after different incubation periods, with apparent molecular masses of 20, 35, 43, 50, 70, 100, 116 and 212 kDa. The results obtained suggest an opportunity for exploring this alternative strategy for proteinases production by actinomycetes, using BSG and CSL as economically feasible substrates.

  2. 不同培养基质对金盏菊生长的影响研究%Effects of Different Culture Medium on the Growth of Calendula officinalis

    Institute of Scientific and Technical Information of China (English)

    陈峰; 沈迎春; 刘阳; 李莉

    2014-01-01

    Using different culture medium,using pot Calendula officinalis as experimental material,research the effect of garden soil,mushroom residue,vermiculite and mixed of these three medium as culture medi-um on the growth and development of Calendula officinalis. The test results show that:the different culture medium containing nutrients difference;effect of different culture medium on the growth of Calendula offici-nalis is different;effect of different culture substrates on chlorophyll content,Catalase activity,superoxide dis-mutase(SOD)activity are different. Comprehensive morphological and physiological indexes on the growth of Calendula officinalis can be concluded:the mushroom residue as growth culture medium of Calendula offici-nalis is best.%采用不同的培养基质,利用盆栽金盏菊作为试验材料,研究田园土、菌渣、蛭石及三者的混合基质作为培养基对金盏菊生长发育的影响。结果表明:不同培养基质所含营养成分差别较大;不同培养基质对金盏菊生长发育影响不同;不同培养基质对金盏菊叶绿素含量、过氧化氢酶活性、超氧化物歧化酶(SOD)活性等影响均不同。综合金盏菊生长的形态指标和生理指标可知:用菌渣作为培养基质的金盏菊长势最好。

  3. The cultural heritage of the metropolitan area of Mendoza (Argentina). Methodological suggestions for their evaluation as a resource in local development project

    OpenAIRE

    Raffa, Cecilia; Conicet Argentina.; Cirvini, Silvia Augusta; Conicet Argentina.

    2014-01-01

    The research methodology proposal as presented, articulate and transfer of applied research results about EnvironmentalHeritage (pca) of the Metropolitan Area of Mendoza (amm). Our general hypothesis argues that the cultural heritage associatedwith the Territory (real estate: architectural, urban, landscape, with its accessories and environment) is a resource and a toolin local development processes, which in the case of the amm is not taken into account by the main actors involved. That is w...

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

    Science.gov (United States)

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

    2017-01-01

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

  5. Effect of low-osmolar contrast medium iopromide and iso-osmolar iodixanol on DNA fragmentation in renal tubular cell culture.

    Science.gov (United States)

    Ludwig, Ulla; Connemann, Julia; Keller, Frieder

    2013-12-01

    Intravascular administration of iodinated contrast media continues to be a common cause of hospital-acquired acute kidney injury. Accumulating evidence suggests that radiocontrast agent-induced nephrotoxicity is associated with increased oxidative stress, which leads to renal tissue damage with DNA fragmentation. We therefore tested whether an iso-osmolar contrast medium (iodixanol) causes less oxidative DNA damage to renal tubular cells than a low-osmolar contrast medium (iopromide). HK-2 cells (human proximal renal tubular cell line) were incubated at different time points (10 min-2 h) with increasing concentrations (20-120 mg/ml iodine) of iodixanol or of iopromide. Oxidative DNA damage to renal tubular cells was measured by alkaline comet assay (single-cell gel electrophoresis). Both iso- and low-osmolar contrast agents induced time- and concentration-dependent DNA fragmentation. DNA fragmentation was maximal at 2 h with 120 mg/ml iodine for iopromide (32 ± 27 tail moments) and iodixanol (46 ± 41 tail moments); both were significantly different from the control value with 3.15 ± 1.6 tail moments (Student's t test; p DNA fragmentation than iopromide (ANOVA for 1 h p = 0.039 and 2 h p = 0.025, respectively). We were able to demonstrate for the first time that an iso-osmolar contrast medium induced even greater oxidative stress and DNA damage than a low-osmolar agent in HK-2 cells. This could provide an explanation for the nephrotoxicity that also is observed with iodixanol in clinical practice.

  6. Internal Control Situation and Suggestions in Small and Medium Enterprises Based on the Idea of Risk Management%基于风险管理理念的中小企业内控现状及改进对策

    Institute of Scientific and Technical Information of China (English)

    徐文静; 陶宗卫

    2011-01-01

    内部控制是实现企业管理目标的过程,它贯穿于企业经营活动的各个方面。企业管理的重要内容之一就是建立风险预警、评估系统,分析各种风险的类型和程度,并采取相应的控制措施。面对日益激烈的市场竞争,中小企业要想立足并不断发展壮大,完善企业的内部控制制度建设,提高企业的风险管理水平,已成为当务之急。%Internal control is a process to realize the management target of a company. It exists in every aspect of enterprise activity. One of important contents in enterprise management is establishment of risk warning and evaluation system to analyze the type and level of the risk and then find appropriate measures. In order to gain a place and grow in the keen market competition, it is urgent for small and medium enterprises to consummate their internal control system step by step and promote its level of risk management.

  7. Immunohistochemical expression of stem cell, endothelial cell, and chemosensitivity markers in primary glioma spheroids cultured in serum-containing and serum-free medium

    DEFF Research Database (Denmark)

    Christensen, Karina; Aaberg-Jessen, Charlotte; Andersen, Claus

    2010-01-01

    To investigate the influence of serum-free medium (SFM) supplemented with epidermal growth factor and basic fibroblast growth factor compared with conventional serum-containing medium (SCM) on the phenotype of organotypic primary spheroids from seven gliomas.......To investigate the influence of serum-free medium (SFM) supplemented with epidermal growth factor and basic fibroblast growth factor compared with conventional serum-containing medium (SCM) on the phenotype of organotypic primary spheroids from seven gliomas....

  8. Optimization of medium composition and cultural conditions for production of antifungal substances by Streptomyces platensis 3-10 and evaluation of its efficacy in suppression of clubroot disease of oilseed rape

    Science.gov (United States)

    This study was conducted to optimize the medium composition and cultural conditions for improving production of antifungal substances (AFS) by Streptomyces 3-10 and for enhancing its efficacy in suppression of clubroot disease of oilseed rape caused by Plasmodiophora brassicae. Results showed that t...

  9. An ultrasensitive LC-MS/MS method with liquid phase extraction to determine paclitaxel in both cell culture medium and lysate promising quantification of drug nanocarriers release in vitro.

    Science.gov (United States)

    Baati, Tarek; Schembri, Thérèse; Villard, Claude; Correard, Florian; Braguer, Diane; Estève, Marie-Anne

    2015-11-10

    The quantification of paclitaxel, a chemotherapy drug used to treat different types of cancers, has been performed from complete cell culture medium and cell lysate samples using a simple liquid-liquid extraction procedure in conjunction with liquid chromatography tandem mass spectrometry (LC-MS/MS). A simple sample preparation using methanol and acetic acid as a weaker acid was applied to avoid paclitaxel destruction and to achieve recovery exceeding 80 % from both matrices spiked with paclitaxel and docetaxel used as internal standard. This rapid, simple, selective and sensitive method enabled the quantification of paclitaxel within the linear range of 1-250nM in culture medium and 5-250nM in cell lysate. The lower limit of quantification was achieved in cell culture medium and cell lysates at 0.2 and 1pmol, respectively. This method was successfully applied to human non-small cell lung carcinoma cells (A549 cells) in order to quantify the amount of paclitaxel in both cell culture medium and lysate after incubation with 5, 50 and 100nM of paclitaxel. This ultra-sensitive method promises the quantification of ultra-low concentrations of paclitaxel released from any nanocarriers, allowing the determination of the kinetic profile of drug release, which is an essential parameter to validate the use of nanocarriers for drug delivery in cancer therapy.

  10. Characterization of the Embryogenic Tissue of the Norway Spruce Including a Transition Layer between the Tissue and the Culture Medium by Magnetic Resonance Imaging

    Science.gov (United States)

    Kořínek, R.; Mikulka, J.; Hřib, J.; Hudec, J.; Havel, L.; Bartušek, K.

    2017-02-01

    The paper describes the visualization of the cells (ESEs) and mucilage (ECMSN) in an embryogenic tissue via magnetic resonance imaging (MRI) relaxometry measurement combined with the subsequent multi-parametric segmentation. The computed relaxometry maps T1 and T2 show a thin layer (transition layer) between the culture medium and the embryogenic tissue. The ESEs, mucilage, and transition layer differ in their relaxation times T1 and T2; thus, these times can be used to characterize the individual parts within the embryogenic tissue. The observed mean values of the relaxation times T1 and T2 of the ESEs, mucilage, and transition layer are as follows: 1469 ± 324 and 53 ± 10 ms, 1784 ± 124 and 74 ± 8 ms, 929 ± 164 and 32 ± 4.7 ms, respectively. The multi-parametric segmentation exploiting the T1 and T2 relaxation times as a classifier shows the distribution of the ESEs and mucilage within the embryogenic tissue. The discussed T1 and T2 indicators can be utilized to characterize both the growth-related changes in an embryogenic tissue and the effect of biotic/abiotic stresses, thus potentially becoming a distinctive indicator of the state of any examined embryogenic tissue.

  11. Effects of granulocyte-macrophage colony-stimulating factor supplementation in culture medium on embryo quality and pregnancy outcome of women aged over 35 years.

    Science.gov (United States)

    Zhou, Wenhui; Chu, Dapeng; Sha, Wei; Fu, Lei; Li, Yuan

    2016-01-01

    The purpose of this study is to explore whether a low concentration of granulocyte-macrophage colony-stimulating factor (GM-CSF) supplementation in culture medium is beneficial to infertile women aged over 35 years. A retrospective cohort study was performed to analyze the embryo quality and pregnancy outcome of 212 controlled ovarian stimulation (COH) cycles with or without GM-CSF addition (n = 117 [GM-CSF, 0.2 ng/mL] vs n = 95 [control]). No significant difference was observed in cleavage rate (96.2 vs 96.5 %), blastocyst formation rate (53.2 vs 54.0 %), good blastocyst rate (26.8 vs 26.8 %), or available embryo rate (54.2 vs 49.7 %) between the GM-CSF group and the control group. However, the average age of the GM-CSF group (38.41 ± 3.13 years) was significantly 1 year older than that of the corresponding control group (37.45 ± 2.74 years) (P quality was observed, the addition of this factor significantly decreased the occurrence of chemical pregnancy of women aged over 35 years, indicating the role of GM-CSF in improving implantation competence of embryos derived from elderly infertile women.

  12. Uranium(VI) complexation in cell culture medium: influence of speciation on Normal Rat Kidney (NRK-52{sup E}) cell accumulation

    Energy Technology Data Exchange (ETDEWEB)

    Carriere, M.; Khodja, H.; Avoscan, L.; Carrot, F.; Gouget, B. [Lab. Pierre Suee CEA-CNRS UMR 9956, Gif sur Yvette (France)

    2005-07-01

    Uranium bioavailability and toxicity are closely linked to the metal's speciation in solution. However in biological fluids or in media classically used for cell culture - and subsequently for in vitro cell exposure -, uranium is rarely present as free-ion since these media contain non-negligible concentrations of potential ligands such as phosphate and bicarbonate but also co-ions such as calcium which can cause U(VI) complexes precipitation. The chemical form of uranium that is internalized in cells and interferes with biological processes is of major concern. Uranium toxicity and accumulation were evaluated in vitro on NRK-52{sup E} cells, model for rat renal proximal tubule. Uranium intracellular accumulation begins after 12 h exposure to 600 {mu}M U(VI); toxicity appears as soon as cells accumulated 25 to 30 mg U/g protein. Modification of uranium speciation in the exposure medium induces great changes in toxicity and cell accumulation. Comparison of toxicity and accumulation results to theoretical uranium speciation, calculated with the J-Chess computer program, shows that free-ion concentration can not explain the total uranium intracellular accumulation. Low molecular weight U(VI) complexes, such as UO{sub 2}(CO{sub 3}){sub 3}{sup 4-} but also UO{sub 2}PO{sub 4}{sup -} could be implicated in U(VI) cellular accumulation and toxicity. (orig.)

  13. Suggested guidelines for using systemic antimicrobials in bacterial skin infections: part 1—diagnosis based on clinical presentation, cytology and culture

    Science.gov (United States)

    Beco, L.; Guaguère, E.; Méndez, C. Lorente; Noli, C.; Nuttall, T.; Vroom, M.

    2013-01-01

    Systemic antimicrobials are critically important in veterinary healthcare, and resistance is a major concern. Antimicrobial stewardship will be important in maintaining clinical efficacy by reducing the development and spread of antimicrobial resistance. Bacterial skin infections are one of the most common reasons for using systemic antimicrobials in dogs and cats. Appropriate management of these infections is, therefore, crucial in any policy for responsible antimicrobial use. The goals of therapy are to confirm that an infection is present, identify the causative bacteria, select the most appropriate antimicrobial, ensure that the infection is treated correctly, and to identify and manage any underlying conditions. This is the first of two articles that will provide evidence-led guidelines to help practitioners address these issues. This article covers diagnosis, including descriptions of the different clinical presentations of surface, superficial and deep bacterial skin infections, how to perform and interpret cytology, and how to best use bacterial culture and sensitivity testing. Part 2 will discuss therapy, including choice of drug and treatment regimens. PMID:23292951

  14. Concise review: no breakthroughs for human mesenchymal and embryonic stem cell culture: conditioned medium, feeder layer, or feeder-free; medium with fetal calf serum, human serum, or enriched plasma; serum-free, serum replacement nonconditioned medium, or ad hoc formula? All that glitters is not gold!

    Science.gov (United States)

    Mannello, Ferdinando; Tonti, Gaetana A

    2007-07-01

    The choice of an optimal strategy of stem cell culture is at the moment an impossible task, and the elaboration of a culture medium adapted to the production of embryonic and adult mesenchymal stem cells for the clinical application of cell therapy remains a crucial matter. To make an informed choice, it is crucial to not underestimate the theoretical health risk of using xenogenic compounds, to limit the immunological reactions once stem cells are transplanted, to not overestimate the controversial results obtained with human serum, plasma, and blood derivatives, as well as to carefully examine the pros and cons of serum-free and ad hoc formulation strategies; besides that, to also maintain multipotentiality, self-renewal, and transplantability. The extent to which we are able to achieve effective cell therapies will depend on assimilating a rapidly developing base of scientific knowledge with the practical considerations of design, delivery, and host response. Although clinical studies have already started, many questions remain unsolved, and concomitantly even more evidence on suitable and safe off-the-shelf products (mainly xeno-free) for embryonic and mesenchymal stem cells is cropping up, even though there should be no rush to enter the clinical stage while the underlying basic research is still not so solid; this solely will lead to high-quality translational research, without making blunders stemming from the assumption that all that glitters is not gold. Disclosure of potential conflicts of interest is found at the end of this article.

  15. Experiments on tissue culture in the genus Lycopersicon miller : Shoot formation from protoplasts of tomato long-term cell cultures.

    Science.gov (United States)

    Koblitz, H; Koblitz, D

    1982-06-01

    Callus cultures from cotyledon explants were established and maintained in culture for more than two years. After several months callus cultures were transferred into liquid medium and cultured as cell suspensions. Protoplasts were isolated from these cell suspension cultures and cultured in a liquid medium. After formation of new cell walls the cells were further cultured in liquid medium and afterwards transferred to an agar-solidified medium to give a vigorously growing callus culture. In the case of the cultivar 'Lukullus' shoots were recovered from callus. All efforts to root these shoots failed and this, in addition to variations in appearence, suggests that the shoots are changed genetically possibly due to the prolonged culture period.

  16. RNA-Based Stable Isotope Probing Suggests Allobaculum spp. as Particularly Active Glucose Assimilators in a Complex Murine Microbiota Cultured In Vitro

    Science.gov (United States)

    Herrmann, Elena; Young, Wayne; Rosendale, Douglas; Reichert-Grimm, Verena; Conrad, Ralf

    2017-01-01

    RNA-based stable isotope probing (RNA-SIP) and metabolic profiling were used to detect actively glucose-consuming bacteria in a complex microbial community obtained from a murine model system. A faeces-derived microbiota was incubated under anaerobic conditions for 0, 2, and 4 h with 40 mM [U13C]glucose. Isopycnic density gradient ultracentrifugation and fractionation of isolated RNA into labeled and unlabeled fractions followed by 16S rRNA sequencing showed a quick adaptation of the bacterial community in response to the added sugar, which was dominated by unclassified Lachnospiraceae species. Inspection of distinct fractions of isotope-labeled RNA revealed Allobaculum spp. as particularly active glucose utilizers in the system, as the corresponding RNA showed significantly higher proportions among the labeled RNA. With time, the labeled sugar was used by a wider spectrum of faecal bacteria. Metabolic profiling indicated rapid fermentation of [U13C]glucose, with lactate, acetate, and propionate being the principal 13C-labeled fermentation products, and suggested that “cross-feeding” occurred in the system. RNA-SIP combined with metabolic profiling of 13C-labeled products allowed insights into the microbial assimilation of a general model substrate, demonstrating the appropriateness of this technology to study assimilation processes of nutritionally more relevant substrates, for example, prebiotic carbohydrates, in the gut microbiota of mice as a model system. PMID:28299315

  17. Induction of E-cadherin+ human amniotic fluid cell differentiation into oocyte-like cells via culture in medium supplemented with follicular fluid.

    Science.gov (United States)

    Liu, Te; Huang, Yongyi; Bu, Yanzhen; Zhao, Yanhui; Zou, Gang; Liu, Zhixue

    2014-07-01

    Pluripotent human amniotic fluid cells (HuAFCs) can differentiate into various types of somatic cell in vitro. However, their differentiation into oocyte-like cells has never been described to the best of our knowledge. In the present study, differentiation of E-cadherin+ and E-cadherin- HuAFC sub-populations into oocyte-like cells was induced via culture in medium containing bovine follicular fluid and β-mercaptoethanol. The E-cadherin+ HuAFCs expressed DAZL highly. Post-induction, cells with an oocyte-like phenotype were found among the E-cadherin+ HuAFCs, expressing markers specific to germ cells and oocytes (VASA, ZP3 and GDF9) and meiosis (DMC1 and SCP3). When specific small interfering RNA (siRNA) was used to suppress E-cadherin in the E-cadherin+ HuAFCs, the levels of DAZL expression were reduced. Post-induction, the morphology of the siRNA‑E‑cadherin HuAFCs was poorer and the expression levels of germ cell-specific markers were lower compared with those of the siRNA-mock HuAFCs. Therefore, E-cadherin+ HuAFCs could be more easily induced to differentiate into oocyte-like cells by bovine follicular fluid and β-mercaptoethanol. In addition, the E-cadherin+ HuAFCs exhibited potential characteristics of DAZL protein expression, and thus it was conjectured that bovine follicular fluid acts on DAZL protein and promotes E-cadherin+ HuAFC differentiation into oocyte-like cells.

  18. Effective clinical-scale production of dendritic cell vaccines by monocyte elutriation directly in medium, subsequent culture in bags and final antigen loading using peptides or RNA transfection.

    Science.gov (United States)

    Erdmann, Michael; Dörrie, Jan; Schaft, Niels; Strasser, Erwin; Hendelmeier, Martin; Kämpgen, Eckhart; Schuler, Gerold; Schuler-Thurner, Beatrice

    2007-09-01

    Dendritic cell (DC) vaccination approaches are advancing fast into the clinic. The major obstacle for further improvement is the current lack of a simple functionally "closed" system to generate standardized monocyte-derived (mo) DC vaccines. Here, we significantly optimized the use of the Elutra counterflow elutriation system to enrich monocytic DC precursors by (1) developing an algorithm to avoid red blood cell debulking and associated monocyte loss before elutriation, and (2) by elutriation directly in culture medium rather than phosphate-buffered saline. Upon elutriation the bags containing the collected monocytes are simply transferred into the incubator to generate DC progeny as the final "open" washing step is no longer required. Elutriation resulted in significantly more (> or = 2-fold) and purer DC than the standard gradient centrifugation/adherence-based monocyte enrichment, whereas morphology, maturation markers, viability, migratory capacity, and T cell stimulatory capacity were identical. Subsequently, we compared RNA transfection, as this is an increasingly used approach to load DC with antigen. Elutra-derived and adherence-derived DC could be electroporated with similar, high efficiency (on average >85% green fluorescence protein positive), and appeared also equal in antigen expression kinetics. Both Elutra-derived and adherence-derived DC, when loaded with the MelanA peptide or electroporated with MelanA RNA, showed a high T cell stimulation capacity, that is, priming of MelanA-specific CD8+ T cells. Our optimized Elutra-based procedure is straightforward, clearly superior to the standard gradient centrifugation/plastic adherence protocol, and now allows the generation of large numbers of peptide-loaded or RNA-transfected DC in a functionally closed system.

  19. Identification of candidate agents active against N. ceranae infection in honey bees: establishment of a medium throughput screening assay based on N. ceranae infected cultured cells.

    Science.gov (United States)

    Gisder, Sebastian; Genersch, Elke

    2015-01-01

    Many flowering plants in both natural ecosytems and agriculture are dependent on insect pollination for fruit set and seed production. Managed honey bees (Apis mellifera) and wild bees are key pollinators providing this indispensable eco- and agrosystem service. Like all other organisms, bees are attacked by numerous pathogens and parasites. Nosema apis is a honey bee pathogenic microsporidium which is widely distributed in honey bee populations without causing much harm. Its congener Nosema ceranae was originally described as pathogen of the Eastern honey bee (Apis cerana) but jumped host from A. cerana to A. mellifera about 20 years ago and spilled over from A. mellifera to Bombus spp. quite recently. N. ceranae is now considered a deadly emerging parasite of both Western honey bees and bumblebees. Hence, novel and sustainable treatment strategies against N. ceranae are urgently needed to protect honey and wild bees. We here present the development of an in vitro medium throughput screening assay for the identification of candidate agents active against N. ceranae infections. This novel assay is based on our recently developed cell culture model for N. ceranae and coupled with an RT-PCR-ELISA protocol for quantification of N. ceranae in infected cells. The assay has been adapted to the 96-well microplate format to allow automated analysis. Several substances with known (fumagillin) or presumed (surfactin) or no (paromomycin) activity against N. ceranae were tested as well as substances for which no data concerning N. ceranae inhibition existed. While fumagillin and two nitroimidazoles (metronidazole, tinidazole) totally inhibited N. ceranae proliferation, all other test substances were inactive. In summary, the assay proved suitable for substance screening and demonstrated the activity of two synthetic antibiotics against N. ceranae.

  20. Growing need to study foraminifera in the laboratory culture experiments: An attempt from the central west coast of India

    Digital Repository Service at National Institute of Oceanography (India)

    Nigam, R.; Khare, N.; Koli, N.Y.

    culture programme is studied. Initially, the effect of different media (like plain sea water and Erdschreiber medium), food and antibiotic drug on the growth of foraminiferal tests are monitored. The results suggest that Erdschreiber medium is condusive...

  1. Zastosowanie peptonu Peptobak-Bacutil tu hodowli merystematycznej tkanki Cymbidium Sw. [Application of the peptone Peptobak-Bacutil medium in the culture of meristem-tissue of Cymbidium Sw.

    Directory of Open Access Journals (Sweden)

    K. Kukułczanka

    2015-06-01

    Full Text Available In was found that peptone of Peptobak-Bacutil brand (produced in Poland fulfills perfectly the requirements in cultures in vitro of the meristem tissues of Cymbidium in the modified Tsuchiya's medium in a concentration of 2 g per 1. A decrease in the concentration to 1 g per 1. affects the growth intensity of the tissue, but an increase of concentration up to 3 g per 1. retards the transformation of protocorms into the plant-stage. Growth of the meristem tissue of Cymbidium in the medium consolidated with agar is weaker and the number of protocorms produced 2,3 times lower than in liquid media.

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

    Directory of Open Access Journals (Sweden)

    Meiyan Xuan

    2015-01-01

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

  3. Pulp fruit added to culture medium for in vitro orchid developmentPolpa de frutos adicionada ao meio de cultivo no crescimento in vitro de orquídea

    Directory of Open Access Journals (Sweden)

    Gilberto Rostirolla Batista de Souza

    2013-06-01

    Full Text Available As an additive in in vitro culture media, fruits have a great potential for facilitating economical orchid production because of lower technology requirements and the ease of obtaining raw materials to formulate culture media. We studied the in vitro growth of Cattleya bicolor Lindl. grown in a simplified culture medium supplemented with different kinds of fruit pulp. The experimental design was completely randomised, with eight seedlings per replication and ten replications per treatment, for a total of 80 seedlings per treatment. The culture medium was made using 150 g L -1 of pulp (without peel or seed from the following fruits: ripe Santa Cruz tomatoes (Solanum lycopersicum L., dwarf bananas (Musa cavendishii L. of intermediate ripeness, light green chayote (Sechium edule (Jacq. Sw, ripe papaya (Carica papaya L. or green coconut (Cocos nucifera L..The treatment control was MS 50 %. The treatments and the control were kept in a growth chamber for seven months before evaluating seedling survival percentage, shoot height, number of leaves, rooting percentage, root number, root length and dry masses of shoot and roots. The highest percentages of seedling survival were obtained using MS 50 %, banana and coconut medium. The seedling survival and rooting percentages illustrate that it is possible to emphasise the culture medium MS 50% and the culture medium supplemented with coconut on the most traditional culture medium with banana or tomato pulp. For the in vitro development of Cattleya bicolor Lindl., a simplified culture medium supplemented with coconut pulp is the most suitable for use as an alternative to MS 50%. A simplified culture medium supplemented with papaya pulp is not recommended for the in vitro development of Cattleya bicolor Lindl. Os frutos apresentam potencial para serem utilizados na elaboração de meios de cultivo para facilitar a produção de orquídeas em pequenas propriedades, contribuindo para a rentabilidade do cultivo

  4. CLINICAL COMPARISON OF GRAFTS STORED IN MCCAREY-KAUFMAN MEDIUM AT 4-DEGREES-C AND IN CORNEAL ORGAN-CULTURE AT 31-DEGREES-C

    NARCIS (Netherlands)

    RIJNEVELD, WJ; BEEKHUIS, WH; VANRIJ, G; RINKELVANDRIEL, B; PELS, E

    Twenty-eight paired human corneas were preserved in minimal essential medium at 31-degrees-C and in McCarey-Kaufman medium at 4-degrees-C. These grafts were then transplanted in pairs of patients with keratoconus who were age matched as closely as possible. These pairs received donor corneas from

  5. EFFECT OF DONOR CORNEA PRESERVATION IN TISSUE-CULTURE AND IN MCCAREY-KAUFMANN MEDIUM ON CORNEAL GRAFT-REJECTION AND VISUAL-ACUITY

    NARCIS (Netherlands)

    MOLL, AC; VANRIJ, G; BEEKHUIS, WH; DELAVALETTE, JHCR; HERMANS, J; PELS, E; RINKELVANDRIEL, E

    1991-01-01

    In a retrospective study the rate of rejection and clouding of the donor cornea for other reasons was investigated in 230 penetrating keratoplasties performed between 1984-1986. Donor corneas were stored in McCarey-Kaufmann Medium (MK) at 4-degrees-C or in a modified Minimal Essential Medium (MEM)

  6. Utilização de resíduos agroindustriais para a produção de proteases pelo termofílico Bacillus sp em fermentação submersa: otimização do meio de cultura usando a técnica de planejamento experimental Protease production using agroindustrial residues by thermophilic Bacillus sp in submerged fermentation: optimization of the culture medium using an experimental design approach

    Directory of Open Access Journals (Sweden)

    Silvania Alves Ladeira

    2010-01-01

    Full Text Available Thermophilic Bacillus sp. SMIA-2, produced protease when grown on apple pectic, whey protein and corn step liquor medium, whose concentration was varied from 3 to 10 gL-1, according to the central composite design 2³. The experiments were conducted in shaker, at 50 °C, 150 rpm and initial pH 6.5. The results revealed that the culture medium affected both, cell growth and enzyme production. After graphical and numerical optimization procedure, the enzyme production reached its maximum value at 30 h fermentation, reaching, approximately, 70 U protein mg-1, suggesting that this process was partially associated to the growth.

  7. 在复合成分培养基中骨髓间充质干细胞的诱导成骨%Osteogenic induction of human bone marrow mesenchymal stem cells cultured in complex medium

    Institute of Scientific and Technical Information of China (English)

    徐丽丽; 孙晓娟; 郝秀仙; 谢婷婷; 杨乃龙

    2015-01-01

    背景:研究显示骨质疏松多伴有成骨细胞的减少,成骨细胞替代疗法成为治疗骨质疏松症的新靶点。目的:观察人骨髓间充质干细胞在含地塞米松、维生素C及β-甘油磷酸钠培养基中向成骨细胞分化的能力。方法:采用人淋巴细胞分离液从成人骨髓血中分离纯化间充质干细胞,应用流式细胞仪检测其表面标志物的活性,透射电镜观察骨髓间充质干细胞的超微结构;将骨髓间充质干细胞在含有地塞米松、维生素 C 与β-甘油磷酸钠的成骨诱导培养基中诱导分化, RT-PCR检测骨髓间充质干细胞成骨诱导后骨形态发生蛋白2 mRNA的表达情况。结果与结论:培养2周后可见细胞呈成纤维状生长,强表达 CD44,CD29,不表达 CD34,CD45,具有向成骨细胞诱导分化的潜能,茜素红及碱性磷酸酶染色阳性,骨形态发生蛋白2 mRNA表达阳性,说明人骨髓间充质干细胞在含地塞米松、维生素C及β-甘油磷酸钠培养基中可向成骨细胞诱导分化,具有治疗骨质疏松症的潜能。%BACKGROUND:Studies have shown that the number of osteoblasts is often decreased after osteoporosis, and osteoblast replacement therapy becomes a new target for the treatment of osteoporosis. OBJECTIVE:To observe the osteogenic differentiation of human bone marrow mesenchymal stem cels cultured in dexamethasone, vitamin C and beta-glycerophosphate. METHODS:Mesenchymal stem cels were isolated and purified from adult bone marrow using human lymphocyte separation medium. The expression of cel surface markers was detected by flow cytometry. Cel ultrastructure was observed by transmission electron microscope. Then, the bone marrow mesenchymal stem cels were cultured in osteogenic induction medium containing dexamethasone, vitamin C andβ-glycerophosphate, and RT-PCR was used to detect the bone morphogenetic protein-2 mRNA expression after osteogenic induction. RESULTS AND

  8. Ways and suggestions for the class cultural construction of modern power supply enterprises%试论现代供电企业班组文化建设的途径与建议

    Institute of Scientific and Technical Information of China (English)

    夏传龙

    2013-01-01

    Paying more attention to the team cultural construction plays an important role for the healthy development of the modern power supply enterprises.By analysing the present situation of team cultural construction in the power supply company where the anthor works,this paper concludes that the team cultural construction is effective and can improve the work efficiency.In view of the current situation,it points out the significance of the team cultural construction to the cultural construction of enterprises,discusses the basic ways of team cultural construction of the modern power supply enterprises and puts forward four suggestions.%注重班组文化建设,对于现代供电企业的健康发展起着积极的重要作用。文章以笔者所在的供电公司班组文化现状为个案,通过分析总结与归纳,认为供电公司在班组文化建设方面做出的尝试是有成效的,能提高工作效率。针对当前状况,指明了班组文化建设对企业文化建设的意义,探讨了现代供电企业班组文化建设的基本途径,同时提出了4点建议。

  9. Nuclear-cytoplasmic "tug of war" during cloning: effects of somatic cell nuclei on culture medium preferences of preimplantation cloned mouse embryos

    National Research Council Canada - National Science Library

    Chung, Young Gie; Mann, Mellissa R W; Bartolomei, Marisa S; Latham, Keith E

    2002-01-01

    .... Embryo culture conditions have not been optimized for cloned embryos, and the effects of culture conditions on these early events and the successful initiation of clonal development have not been examined...

  10. Nuclear-Cytoplasmic “Tug of War” During Cloning: Effects of Somatic Cell Nuclei on Culture Medium Preferences of Preimplantation Cloned Mouse Embryos1

    National Research Council Canada - National Science Library

    Young Gie Chung; Mellissa R. W. M