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Sample records for agar-plate culture system

  1. Efficacy of agar-plate culture in detection of Strongyloides stercoralis infection.

    PubMed

    Arakaki, T; Iwanaga, M; Kinjo, F; Saito, A; Asato, R; Ikeshiro, T

    1990-06-01

    Agar-plate culture of feces using a modified petri dish proved to be highly efficient in the detection of Strongyloides stercoralis infection. Furrows left by S. stercoralis on the agar plate were distinguished readily in size from those left by Necator americanus. PMID:2352073

  2. A Chlorhexidine- Agar Plate Culture Medium Protocol to Complement Standard Broth Culture of Mycobacterium tuberculosis

    PubMed Central

    Asmar, Shady; Chatellier, Sonia; Mirande, Caroline; van Belkum, Alex; Canard, Isabelle; Raoult, Didier; Drancourt, Michel

    2016-01-01

    The culture of Mycobacterium tuberculosis using parallel inoculation of a solid culture medium and a liquid broth provides the gold standard for the diagnosis of tuberculosis. Here, we evaluated a chlorhexidine decontamination-MOD9 solid medium protocol versus the standard NALC-NaOH-Bactec 960 MGIT protocol for the diagnosis of pulmonary tuberculosis by culture. Three-hundred clinical specimens comprising 193 sputa, 30 bronchial aspirates, 10 broncho-alveolar lavages, 47 stools, and 20 urines were prospectively submitted for the routine diagnosis of tuberculosis. The contamination rates were 5/300 (1.7%) using the MOD9 protocol and 17/300 (5.7%) with the Bactec protocol, respectively (P < 0.05, Fisher exact test). Of a total of 50 Mycobacterium isolates (48 M. tuberculosis and two Mycobacterium abscessus) were cultured. Out of these 50, 48 (96%) isolates were found using the MOD9 protocol versus 35 (70%) when using the Bactec protocol (P < 0.05, Fisher exact test). The time to positivity was 10.1 ± 3.9 days versus 14.7 ± 7.3 days, respectively, (P < 0.05, Student’s t-test). These data confirmed the usefulness of parallel inoculation of a solid culture medium with broth for the recovery of M. tuberculosis in agreement with current recommendations. More specifically, chlorhexidine decontamination and inoculation of the MOD9 solid medium could be proposed to complement the standard Bactec 960 MGIT broth protocol. PMID:26834733

  3. A Chlorhexidine- Agar Plate Culture Medium Protocol to Complement Standard Broth Culture of Mycobacterium tuberculosis.

    PubMed

    Asmar, Shady; Chatellier, Sonia; Mirande, Caroline; van Belkum, Alex; Canard, Isabelle; Raoult, Didier; Drancourt, Michel

    2016-01-01

    The culture of Mycobacterium tuberculosis using parallel inoculation of a solid culture medium and a liquid broth provides the gold standard for the diagnosis of tuberculosis. Here, we evaluated a chlorhexidine decontamination-MOD9 solid medium protocol versus the standard NALC-NaOH-Bactec 960 MGIT protocol for the diagnosis of pulmonary tuberculosis by culture. Three-hundred clinical specimens comprising 193 sputa, 30 bronchial aspirates, 10 broncho-alveolar lavages, 47 stools, and 20 urines were prospectively submitted for the routine diagnosis of tuberculosis. The contamination rates were 5/300 (1.7%) using the MOD9 protocol and 17/300 (5.7%) with the Bactec protocol, respectively (P < 0.05, Fisher exact test). Of a total of 50 Mycobacterium isolates (48 M. tuberculosis and two Mycobacterium abscessus) were cultured. Out of these 50, 48 (96%) isolates were found using the MOD9 protocol versus 35 (70%) when using the Bactec protocol (P < 0.05, Fisher exact test). The time to positivity was 10.1 ± 3.9 days versus 14.7 ± 7.3 days, respectively, (P < 0.05, Student's t-test). These data confirmed the usefulness of parallel inoculation of a solid culture medium with broth for the recovery of M. tuberculosis in agreement with current recommendations. More specifically, chlorhexidine decontamination and inoculation of the MOD9 solid medium could be proposed to complement the standard Bactec 960 MGIT broth protocol. PMID:26834733

  4. Trace Amounts of Furan-2-Carboxylic Acids Determine the Quality of Solid Agar Plates for Bacterial Culture

    PubMed Central

    Hara, Shintaro; Isoda, Reika; Tahvanainen, Teemu; Hashidoko, Yasuyuki

    2012-01-01

    Background Many investigators have recognised that a significant proportion of environmental bacteria exist in a viable but non-culturable state on agar plates, and some researchers have also noticed that some of such bacteria clearly recover their growth on matrices other than agar. However, the reason why agar is unsuitable for the growth of some bacteria has not been addressed. Methodology/Principal Findings According to the guide of a bioassay for swarming inhibition, we identified 5-hydroxymethylfuran-2-carboxylic acid (5-HMFA) and furan-2-carboxylic acid (FA) as factors that inhibit bacterial swarming and likely inhibit extracellular polysaccharide production on agar. The furan-2-carboxylic acids 5-HMFA and FA effectively inhibited the swarming and swimming of several environmental bacteria at concentrations of 1.8 and 2.3 µg L−1 (13 and 21 nmol L−1), respectively, which are equivalent to the concentrations of these compounds in 0.3% agar. On Luria-Bertani (LB) plates containing 1.0% agar that had been previously washed with MeOH, a mixture of 5-HMFA and FA in amounts equivalent to their original concentrations in the unwashed agar repressed the swarming of Escherichia coli K12 strain W3110, a representative swarming bacterium. Conclusions/Significance Agar that contains trace amounts of 5-HMFA and FA inhibits the proliferation of some slow-growing or difficult-to-culture bacteria on the plates, but it is useful for single colony isolation due to the ease of identification of swarmable bacteria as the non-swarmed colonies. PMID:22848437

  5. Identification of Brucella by MALDI-TOF Mass Spectrometry. Fast and Reliable Identification from Agar Plates and Blood Cultures

    PubMed Central

    Ferreira, Laura; Vega Castaño, Silvia; Sánchez-Juanes, Fernando; González-Cabrero, Sandra; Menegotto, Fabiola; Orduña-Domingo, Antonio

    2010-01-01

    Background MALDI-TOF mass spectrometry (MS) is a reliable method for bacteria identification. Some databases used for this purpose lack reference profiles for Brucella species, which is still an important pathogen in wide areas around the world. We report the creation of profiles for MALDI-TOF Biotyper 2.0 database (Bruker Daltonics, Germany) and their usefulness for identifying brucellae from culture plates and blood cultures. Methodology/Principal Findings We created MALDI Biotyper 2.0 profiles for type strains belonging to B. melitensis biotypes 1, 2 and 3; B. abortus biotypes 1, 2, 5 and 9; B. suis, B. canis, B ceti and B. pinnipedialis. Then, 131 clinical isolates grown on plate cultures were used in triplicate to check identification. Identification at genus level was always correct, although in most cases the three replicates reported different identification at species level. Simulated blood cultures were performed with type strains belonging to the main human pathogenic species (B. melitensis, B. abortus, B. suis and B. canis), and studied by MALDI-TOF MS in triplicate. Identification at genus level was always correct. Conclusions/Significance MALDI-TOF MS is reliable for Brucella identification to the genus level from culture plates and directly from blood culture bottles. PMID:21151913

  6. Hyperspectral imaging for detecting pathogens grown on agar plates

    NASA Astrophysics Data System (ADS)

    Yoon, Seung Chul; Lawrence, Kurt C.; Siragusa, Gregory R.; Line, John E.; Park, Bosoon; Windham, William R.

    2007-09-01

    This paper is concerned with the development of a hyperspectral imaging technique for detecting and identifying one of the most common foodborne pathogens, Campylobacter. Direct plating using agars is an effective tool for laboratory tests and analyses of microorganisms. The morphology (size, growth pattern, color, etc.) of colonies grown on agar plates has been widely used to tentatively differentiate organisms. However, it is sometimes difficult to differentiate target organisms like Campylobacters from other contaminants grown together on the same agar plates. A hyperspectral imaging system operating at the visible and near infrared (VNIR) spectral region from 400 nm to 900 nm was set up to measure spectral signatures of 17 different Campylobacter and non-Campylobacter subspecies. Protocols for culturing, imaging samples and for calibrating measured data were developed. The VNIR spectral library of all 17 organisms commonly encountered in poultry was established from calibrated hyperspectral images. A classification algorithm was developed to locate and identify Campylobacters, non-Campylobacter contaminants, and background agars with 99.29% accuracy. This research has a potential to be expanded to detect other pathogens grown on agar media.

  7. Simplified 48-hour IMVic test: an agar plate method.

    PubMed

    Powers, E M; Latt, T G

    1977-09-01

    An agar plate method was developed for the performance of the IMVic (indole, methyl red, Voges-Proskauer, and citrate) tests in lieu of the conventional tubed liquid media. By modifying the composition of the media and adding agar, a single "X"-compartmented petri dish was prepared containing all four IMVic test media. Ease of performance and simplification of the test were achieved by inoculating all four media simultaneously from a single colony (single inoculum) on eosin-methylene blue agar. Tests with 87 cultures, representing 7 genera in the family Enterobacteriaceae, were completed with typical (correct) IMVic patterns for all cultures within 48 h. Parallel tests with conventional media showed that the agar plate method was superior, more sensitive, faster, and simpler to perform, and less time was required to identify Escherichia coli by 72 h. PMID:334074

  8. Syneresis and delayed detachment in agar plates.

    PubMed

    Divoux, Thibaut; Mao, Bosi; Snabre, Patrick

    2015-05-14

    Biogels made of crosslinked polymers such as proteins or polysaccharides behave as porous soft solids and store large amounts of solvent. These gels undergo spontaneous aging, called syneresis, which consists of the shrinkage of the gel matrix and the progressive expulsion of solvent. As a result, a biogel originally casted in a container often loses contact with the container sidewalls, and the detachment time is difficult to anticipate a priori, since it may occur over variable time spans (from hours to days). Here we report on syneresis phenomena in agar plates, which consist of Petri dishes filled with a gel mainly composed of agar. Direct observations and speckle pattern correlation analysis allow us to rationalize the delayed detachment of the gel from the sidewall of the Petri dish. The detachment time t* is surprisingly not controlled by the mass loss as one would intuitively expect. Instead, t* is strongly correlated to the gel minimum thickness emin measured along the sidewall of the plate, and increases as a robust function of emin, independently of the prior mass-loss history. Time-resolved correlation spectroscopy atypically applied to such weakly diffusive media gives access to the local thinning rate of the gel. This technique also allows us to detect the gel micro-displacements that are triggered by water evaporation prior to the detachment, and even to anticipate the latter from a few hours. Our work provides observables to predict the detachment time of agar gels in dishes, and highlights the relevance of speckle pattern correlation analysis for the quantitative investigation of the syneresis dynamics in biopolymer gels. PMID:25812667

  9. 21 CFR 866.4600 - Ouchterlony agar plate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Ouchterlony agar plate. 866.4600 Section 866.4600 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunology Laboratory Equipment and Reagents §...

  10. 21 CFR 866.4600 - Ouchterlony agar plate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Ouchterlony agar plate. 866.4600 Section 866.4600 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunology Laboratory Equipment and Reagents §...

  11. 21 CFR 866.4600 - Ouchterlony agar plate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Ouchterlony agar plate. 866.4600 Section 866.4600 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunology Laboratory Equipment and Reagents §...

  12. 21 CFR 866.4600 - Ouchterlony agar plate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Ouchterlony agar plate. 866.4600 Section 866.4600 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunology Laboratory Equipment and Reagents §...

  13. 21 CFR 866.4600 - Ouchterlony agar plate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Ouchterlony agar plate. 866.4600 Section 866.4600 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunology Laboratory Equipment and Reagents §...

  14. A modified agar plate method for detection of Strongyloides stercoralis.

    PubMed

    Koga, K; Kasuya, S; Khamboonruang, C; Sukhavat, K; Ieda, M; Takatsuka, N; Kita, K; Ohtomo, H

    1991-10-01

    The agar plate method is a new technique with high detection rates for coprological diagnosis of human strongyloidiasis. This report details modifications of the technique and establishes a standardized procedure. We recommend that all plates should be carefully observed using a microscope because macroscopic observation can lead to false negative results. It is also advisable to pour formalin solution directly into microscopically positive dishes to collect worms by sedimentation. This procedure enables one to observe worms otherwise hidden. Sealing dishes with adhesive tape prevents larvae from crawling out of the dishes, eliminating any possibility in the reduction of detection rates, and greatly improves the safety conditions for the technician performing the procedure. We consider the agar plate method to be superior to the filter paper method in detecting Strongyloides, and we believe that it will eventually become the technique of choice. PMID:1951861

  15. Individual based simulations of bacterial growth on agar plates

    NASA Astrophysics Data System (ADS)

    Ginovart, M.; López, D.; Valls, J.; Silbert, M.

    2002-03-01

    The individual based simulator, INDividual DIScrete SIMulations (INDISIM) has been used to study the behaviour of the growth of bacterial colonies on a finite dish. The simulations reproduce the qualitative trends of pattern formation that appear during the growth of Bacillus subtilis on an agar plate under different initial conditions of nutrient peptone concentration, the amount of agar on the plate, and the temperature. The simulations are carried out by imposing closed boundary conditions on a square lattice divided into square spatial cells. The simulator studies the temporal evolution of the bacterial population possible by setting rules of behaviour for each bacterium, such as its uptake, metabolism and reproduction, as well as rules for the medium in which the bacterial cells grow, such as concentration of nutrient particles and their diffusion. The determining factors that characterize the structure of the bacterial colony patterns in the presents simulations, are the initial concentrations of nutrient particles, that mimic the amount of peptone in the experiments, and the set of values for the microscopic diffusion parameter related, in the experiments, to the amount of the agar medium.

  16. Mass production of spores of lactic acid-producing Rhizopus oryzae NBRC 5384 on agar plate.

    PubMed

    Yamane, Tsuneo; Tanaka, Ryosuke

    2013-01-01

    Mass production of sporangiospores (spores) of Rhizopus oryzae NBRC 5384 (identical to NRRL 395 and ATCC 9363) on potato-dextrose-agar medium was studied aiming at starting its L(+)-lactic acid fermentation directly from spore inoculation. Various parameters including harvest time, sowed spore density, size of agar plate, height of air space, and incubation mode of plate (agar-on-bottom or agar-on-top) were studied. Ordinarily used shallow Petri dishes were found out to be unsuitable for the full growth of R. oryzae sporangiophores. In a very wide range of the sowed spore density, the smaller it was, the greater the number of the harvested spores was. It was also interesting to find out that R. oryzae grown downward vertically with a deep air space in an agar-on-top mode gave larger amount of spores than in an agar-on-bottom mode at 30°C for 7-day cultivation. Scale-up of the agar plate culture from 26.4 to 292 cm(2) was studied, resulting in the proportional relationship between the number of the harvested spores/plate and the plate area in the deep Petri dishes. The number of plates of 50 cm in diameter needed for 100 m(3) industrial submerged fermentation started directly from 2 × 10(5) spores/mL inoculum size was estimated as about 6, from which it was inferred that such a fermentation would be feasible. Designing a 50 cm plate and a method of spreading and collecting the spores were suggested. Bioprocess technological significance of the "full-scale industrial submerged fermentation started directly from spore inoculation omitting pre-culture" has been discussed. PMID:23658025

  17. Development of a colony lift immunoassay to facilitate rapid detection and quantification of Escherichia coli O157:H7 from agar plates and filter monitor membranes.

    PubMed

    Ingram, D T; Lamichhane, C M; Rollins, D M; Carr, L E; Mallinson, E T; Joseph, S W

    1998-07-01

    E. coli O157:H7 is a food-borne adulterant that can cause hemorrhagic ulcerative colitis and hemolytic uremic syndrome. Faced with an increasing risk of foods contaminated with E. coli O157:H7, food safety officials are seeking improved methods to detect and isolate E. coli O157:H7 in hazard analysis and critical control point systems in meat- and poultry-processing plants. A colony lift immunoassay was developed to facilitate the positive identification and quantification of E. coli O157:H7 by incorporating a simple colony lift enzyme-linked immunosorbent assay with filter monitors and traditional culture methods. Polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, Mass.) were prewet with methanol and were used to make replicates of every bacterial colony on agar plates or filter monitor membranes that were then reincubated for 15 to 18 h at 36 +/- 1 degree C, during which the colonies not only remained viable but were reestablished. The membranes were dried, blocked with blocking buffer (Kirkegaard and Perry Laboratories [KPL], Gaithersburg, Md.), and exposed for 7 min to an affinity-purified horseradish peroxidase-labeled goat anti-E. coli O157 antibody (KPL). The membranes were washed, exposed to a 3,3',5,5'-tetramethylbenzidine membrane substrate (TMB; KPL) or aminoethyl carbazole (AEC; Sigma Chemical Co., St. Louis, Mo.), rinsed in deionized water, and air dried. Colonies of E. coli O157:H7 were identified by either a blue (via TMB) or a red (via AEC) color reaction. The colored spots on the PVDF lift membrane were then matched to their respective parent colonies on the agar plates or filter monitor membranes. The colony lift immunoassay was tested with a wide range of genera in the family Enterobacteriaceae as well as different serotypes within the E. coli genus. The colony lift immunoassay provided a simple, rapid, and accurate method for confirming the presence of E. coli O157:H7 colonies isolated on filter monitors or spread plates by

  18. Light‐scattering sensor for real‐time identification of Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae colonies on solid agar plate

    PubMed Central

    Huff, Karleigh; Aroonnual, Amornrat; Littlejohn, Amy E. Fleishman; Rajwa, Bartek; Bae, Euiwon; Banada, Padmapriya P.; Patsekin, Valery; Hirleman, E. Daniel; Robinson, J. Paul; Richards, Gary P.; Bhunia, Arun K.

    2012-01-01

    Summary The three most common pathogenic species of Vibrio, Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus, are of major concerns due to increased incidence of water‐ and seafood‐related outbreaks and illness worldwide. Current methods are lengthy and require biochemical and molecular confirmation. A novel label‐free forward light‐scattering sensor was developed to detect and identify colonies of these three pathogens in real time in the presence of other vibrios in food or water samples. Vibrio colonies grown on agar plates were illuminated by a 635 nm laser beam and scatter‐image signatures were acquired using a CCD (charge‐coupled device) camera in an automated BARDOT (BActerial Rapid Detection using Optical light‐scattering Technology) system. Although a limited number of Vibrio species was tested, each produced a unique light‐scattering signature that is consistent from colony to colony. Subsequently a pattern recognition system analysing the collected light‐scatter information provided classification in 1−2 min with an accuracy of 99%. The light‐scattering signatures were unaffected by subjecting the bacteria to physiological stressors: osmotic imbalance, acid, heat and recovery from a viable but non‐culturable state. Furthermore, employing a standard sample enrichment in alkaline peptone water for 6 h followed by plating on selective thiosulphate citrate bile salts sucrose agar at 30°C for ∼ 12 h, the light‐scattering sensor successfully detected V. cholerae, V. parahaemolyticus and V. vulnificus present in oyster or water samples in 18 h even in the presence of other vibrios or other bacteria, indicating the suitability of the sensor as a powerful screening tool for pathogens on agar plates. PMID:22613192

  19. Agar Plate Method for Detection and Enumeration of Alkylbenzenesulfonate-Degrading Microorganisms

    PubMed Central

    Ohwada, Kouichi

    1975-01-01

    A simple method for detection and enumeration of alkylbenzenesulfonate (ABS)-degrading microorganisms by using agar plates was developed and used in microbiological studies of coastal marine and polluted river waters. The method depends upon the color responses of neutral red in alkaline medium. Neutral red changes from pink, when it enters into ABS micelles, to yellow, when the ABS is degraded, and does not form micelles. When neutral red-tris(hydroxymethyl)-aminomethane buffer solution and then cationic surfactant solution were sprayed onto the agar surface of ABS-nutrient agar cultures, transparent haloes appeared around the colonies of ABS-degrading microorganisms against a pink background. Viable counts of ABS-degrading bacteria isolated from both seawater and freshwater environments were considerably higher in polluted waters than in less polluted areas. Viable counts of ABS-degrading bacteria averaged 1.5 × 105/ml in samples from the surface water of polluted Tokyo Bay and 3.0 × 104/ml in samples from the surface water of polluted Tamagawa River but were fewer in number in samples from less polluted waters. Images PMID:234155

  20. Development of hyperspectral imaging technique for salmonella enteritidis and typhimurium on agar plates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella is a common cause of foodborne disease resulting from the consumption of contaminated food products. Although a direct plating method is widely used for presumptive positive screening of pathogenic Salmonella colonies on agar plates, it is labor-intensive, time-consuming and also prone to...

  1. Hyperspectral image reconstruction using RGB color for foodborne pathogen detection on agar plates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This paper reports the latest development of a color vision technique for detecting colonies of foodborne pathogens grown on agar plates with a hyperspectral image classification model that was developed using full hyperspectral data. The hyperspectral classification model depended on reflectance sp...

  2. Hyperspectral image reconstruction using RGB color for foodborne pathogen detection on agar plates

    NASA Astrophysics Data System (ADS)

    Yoon, Seung-Chul; Shin, Tae-Sung; Park, Bosoon; Lawrence, Kurt C.; Heitschmidt, Gerald W.

    2014-03-01

    This paper reports the latest development of a color vision technique for detecting colonies of foodborne pathogens grown on agar plates with a hyperspectral image classification model that was developed using full hyperspectral data. The hyperspectral classification model depended on reflectance spectra measured in the visible and near-infrared spectral range from 400 and 1,000 nm (473 narrow spectral bands). Multivariate regression methods were used to estimate and predict hyperspectral data from RGB color values. The six representative non-O157 Shiga-toxin producing Eschetichia coli (STEC) serogroups (O26, O45, O103, O111, O121, and O145) were grown on Rainbow agar plates. A line-scan pushbroom hyperspectral image sensor was used to scan 36 agar plates grown with pure STEC colonies at each plate. The 36 hyperspectral images of the agar plates were divided in half to create training and test sets. The mean Rsquared value for hyperspectral image estimation was about 0.98 in the spectral range between 400 and 700 nm for linear, quadratic and cubic polynomial regression models and the detection accuracy of the hyperspectral image classification model with the principal component analysis and k-nearest neighbors for the test set was up to 92% (99% with the original hyperspectral images). Thus, the results of the study suggested that color-based detection may be viable as a multispectral imaging solution without much loss of prediction accuracy compared to hyperspectral imaging.

  3. Simulation of Bacillus subtilis biofilm growth on agar plate by diffusion–reaction based continuum model

    NASA Astrophysics Data System (ADS)

    Zhang, Xianlong; Wang, Xiaoling; Nie, Kai; Li, Mingpeng; Sun, Qingping

    2016-08-01

    Various species of bacteria form highly organized spatially-structured aggregates known as biofilms. To understand how microenvironments impact biofilm growth dynamics, we propose a diffusion–reaction continuum model to simulate the formation of Bacillus subtilis biofilm on an agar plate. The extended finite element method combined with level set method are employed to perform the simulation, numerical results show the quantitative relationship between colony morphologies and nutrient depletion over time. Considering that the production of polysaccharide in wild-type cells may enhance biofilm spreading on the agar plate, we inoculate mutant colony incapable of producing polysaccharide to verify our results. Predictions of the glutamate source biofilm’s shape parameters agree with the experimental mutant colony better than that of glycerol source biofilm, suggesting that glutamate is rate limiting nutrient for Bacillus subtilis biofilm growth on agar plate, and the diffusion-limited is a better description to the experiment. In addition, we find that the diffusion time scale is of the same magnitude as growth process, and the common-employed quasi-steady approximation is not applicable here.

  4. Simulation of Bacillus subtilis biofilm growth on agar plate by diffusion-reaction based continuum model.

    PubMed

    Zhang, Xianlong; Wang, Xiaoling; Nie, Kai; Li, Mingpeng; Sun, Qingping

    2016-01-01

    Various species of bacteria form highly organized spatially-structured aggregates known as biofilms. To understand how microenvironments impact biofilm growth dynamics, we propose a diffusion-reaction continuum model to simulate the formation of Bacillus subtilis biofilm on an agar plate. The extended finite element method combined with level set method are employed to perform the simulation, numerical results show the quantitative relationship between colony morphologies and nutrient depletion over time. Considering that the production of polysaccharide in wild-type cells may enhance biofilm spreading on the agar plate, we inoculate mutant colony incapable of producing polysaccharide to verify our results. Predictions of the glutamate source biofilm's shape parameters agree with the experimental mutant colony better than that of glycerol source biofilm, suggesting that glutamate is rate limiting nutrient for Bacillus subtilis biofilm growth on agar plate, and the diffusion-limited is a better description to the experiment. In addition, we find that the diffusion time scale is of the same magnitude as growth process, and the common-employed quasi-steady approximation is not applicable here. PMID:27434099

  5. Visualization of Biosurfactant Film Flow in a Bacillus subtilis Swarm Colony on an Agar Plate

    PubMed Central

    Kim, Kyunghoon; Kim, Jung Kyung

    2015-01-01

    Collective bacterial dynamics plays a crucial role in colony development. Although many research groups have studied the behavior of fluidic swarm colonies, the detailed mechanics of its motion remains elusive. Here, we developed a visualization method using submicron fluorescent beads for investigating the flow field in a thin layer of fluid that covers a Bacillus subtilis swarm colony growing on an agar plate. The beads were initially embedded in the agar plate and subsequently distributed spontaneously at the upper surface of the expanding colony. We conducted long-term live cell imaging of the B. subtilis colony using the fluorescent tracers, and obtained high-resolution velocity maps of microscale vortices in the swarm colony using particle image velocimetry. A distinct periodic fluctuation in the average speed and vorticity of flow in swarm colony was observed at the inner region of the colony, and correlated with the switch between bacterial swarming and growth phases. At the advancing edge of the colony, both the magnitudes of velocity and vorticity of flow in swarm colony were inversely correlated with the spreading speed of the swarm edge. The advanced imaging tool developed in this study would facilitate further understanding of the effect of micro vortices in swarm colony on the collective dynamics of bacteria. PMID:26343634

  6. Visualization of Biosurfactant Film Flow in a Bacillus subtilis Swarm Colony on an Agar Plate.

    PubMed

    Kim, Kyunghoon; Kim, Jung Kyung

    2015-01-01

    Collective bacterial dynamics plays a crucial role in colony development. Although many research groups have studied the behavior of fluidic swarm colonies, the detailed mechanics of its motion remains elusive. Here, we developed a visualization method using submicron fluorescent beads for investigating the flow field in a thin layer of fluid that covers a Bacillus subtilis swarm colony growing on an agar plate. The beads were initially embedded in the agar plate and subsequently distributed spontaneously at the upper surface of the expanding colony. We conducted long-term live cell imaging of the B. subtilis colony using the fluorescent tracers, and obtained high-resolution velocity maps of microscale vortices in the swarm colony using particle image velocimetry. A distinct periodic fluctuation in the average speed and vorticity of flow in swarm colony was observed at the inner region of the colony, and correlated with the switch between bacterial swarming and growth phases. At the advancing edge of the colony, both the magnitudes of velocity and vorticity of flow in swarm colony were inversely correlated with the spreading speed of the swarm edge. The advanced imaging tool developed in this study would facilitate further understanding of the effect of micro vortices in swarm colony on the collective dynamics of bacteria. PMID:26343634

  7. Nocardia pigrifrangens sp. nov., a novel actinomycete isolated from a contaminated agar plate.

    PubMed

    Wang, Liming; Zhang, Yamei; Huang, Ying; Maldonado, Luis A; Liu, Zhiheng; Goodfellow, Michael

    2004-09-01

    A polyphasic study was undertaken to establish the taxonomic position of an actinomycete strain isolated from a contaminated agar plate. The strain, designated 7031T, had morphological and chemotaxonomic properties typical of the genus Nocardia. An almost-complete 16S rRNA gene sequence determined for the strain was aligned with available sequences for nocardiae, and phylogenetic trees were inferred using three tree-generating algorithms. Strain 7031T clustered with the type strains of Nocardia carnea and Nocardia flavorosea, showing low 16S rRNA gene sequence similarities to these species (97.2 and 97.5 %, respectively). The strain was also distinguished from the closest species by a range of phenotypic properties. It is proposed that the strain be recognized as a novel species of Nocardia, Nocardia pigrifrangens sp. nov., the type strain of which is 7031T (= AS 4.1808T = JCM 11884T). PMID:15388728

  8. Laboratory detection of extended-spectrum-beta-lactamase-producing Enterobacteriaceae: evaluation of two screening agar plates and two confirmation techniques.

    PubMed

    Overdevest, I T M A; Willemsen, I; Elberts, S; Verhulst, C; Kluytmans, J A J W

    2011-02-01

    The worldwide prevalence of extended-spectrum-beta-lactamase-producing ESBL-producing Enterobacteriaceae (ESBL-E) is increasing, making the need for optimized detection techniques more urgent. In this study we investigated the performance of two ESBL-E screening and two ESBL-E confirmation techniques. In accordance with the Dutch national guidelines (www.wip.nl), a collection of 642 highly resistant Enterobacteriaceae strains, as identified by Vitek2, was used to test the performances of two screening techniques (EbSA ESBL agar plate and ChromID ESBL agar plate) and of two confirmation techniques (MIC-strip ESBL and Vitek2 ESBL test panel). The individual test results were compared by using Etest, followed by a combination disk test if Etest results were inconclusive. Among group 1 isolates (Escherichia coli, Klebsiella spp., Proteus spp., Salmonella spp., and Shigella spp.) 291 (57.6%) were ESBL-E, versus 65 (47.4%) in group 2 (Enterobacter spp., Citrobacter spp., Morganella morganii, Serratia spp., and Providencia spp.). The sensitivities of all four tests for group 1 were comparable (EbSA, 96.6%; ChromID, 97.3%; MIC-strip, 99.6%; and Vitek2, 95.1%). The specificities of the EbSA and ChromID were the same (93.9%). However, the confirmation techniques produced many inconclusive test results, which reduces the applicability in routine laboratories. Only the two screening agar plates were validated for ESBL testing of group 2 microorganisms. They showed comparable sensitivities; however, the EbSA screening agar plate had a significantly higher specificity (78.6% versus 44.3%). In conclusion the screening agar plates performed better than the two confirmation techniques. The EbSA agar plate had the best overall performance. PMID:21123527

  9. Time- and media-saving testing and identification of microorganisms by multipoint inoculation on undivided agar plates.

    PubMed Central

    Burman, L G; Ostensson, R

    1978-01-01

    Motility and various biochemical activities of isolates of bacteria and yeasts were tested on undivided agar plates by using a simple, manually operated multipoint inoculation apparatus that allowed the analysis of 25 isolates per 9-cm-diameter petri plate. Fermentation of all 17 carbohydrates tested as well as 13 other biochemical activities commonly used for identification of bacteria were readily demonstrated by the multipoint inoculation plate method, and the results agreed very well with those of conventional tube tests. In addition to speedy inoculation and low cost of materials, the multipoint inoculation plate method offers several other advantages when compared with conventional tube tests or with some of the manufactured test kits currently available for recognizing members of the family Enterobacteriaceae. Images PMID:359588

  10. Correlation between Agar Plate Screening and Solid-State Fermentation for the Prediction of Cellulase Production by Trichoderma Strains.

    PubMed

    Florencio, Camila; Couri, Sonia; Farinas, Cristiane Sanchez

    2012-01-01

    The viability of converting biomass into biofuels and chemicals still requires further development towards the reduction of the enzyme production costs. Thus, there is a growing demand for the development of efficient procedures for selection of cellulase-producing microorganisms. This work correlates qualitative screening using agar plate assays with quantitative measurements of cellulase production during cultivation under solid-state fermentation (SSF). The initial screening step consisted of observation of the growth of 78 preselected strains of the genus Trichoderma on plates, using microcrystalline cellulose as carbon source. The 49 strains that were able to grow on this substrate were then subjected to a second screening step using the Congo red test. From this test it was possible to select 10 strains that presented the highest enzymatic indices (EI), with values ranging from 1.51 to 1.90. SSF cultivations using sugarcane bagasse and wheat bran as substrates were performed using selected strains. The CG 104NH strain presented the highest EGase activity (25.93 UI·g(-1)). The EI results obtained in the screening procedure using plates were compared with cellulase production under SSF. A correlation coefficient (R(2)) of 0.977 was obtained between the Congo red test and SSF, demonstrating that the two methodologies were in good agreement. PMID:23227312

  11. Direct comparison of the Ames microplate format (MPF) test in liquid medium with the standard Ames pre-incubation assay on agar plates by use of equivocal to weakly positive test compounds.

    PubMed

    Flückiger-Isler, Sini; Kamber, Markus

    2012-08-30

    The Ames microplate format (MPF™) test, which uses liquid media and in 384-well microplates with a readout based on a colour-change, has been used for over 10 years at several major pharmaceutical companies for screening the genotoxic potential of early drug candidates when compound supply is minimal. Meanwhile, Xenometrix has adapted this screen from the two-strain Ames II test for use with five tester strains, in compliance with OECD Guideline 471. A set of 15 equivocal to weakly positive chemicals selected from the National Toxicology Program (NTP) database was tested simultaneously in the Ames microplate format (MPF) and the standard Ames pre-incubation method on agar plates. Such a direct comparison of the two test methods with the same overnight culture(s), chemicals and S9-mix preparation should exclude external variability factors. Thirteen of the 15 chemicals showed concordant results in both tests despite the choice of chemicals that showed varying inter- and even intra-laboratory results in the NTP studies. These results indicate that the Ames MPF™ assay is a reliable predictive tool that can be used like the regular Ames test to evaluate compounds for mutagenicity. PMID:22579797

  12. Production of putative virulence factors by Renibacterium salmoninarum grown in cell culture.

    PubMed

    McIntosh, D; Flaño, E; Grayson, T H; Gilpin, M L; Austin, B; Villena, A J

    1997-10-01

    A cell culture system, employing the fish cell line Epithelioma papillosum cyprini (EPC), was developed to study the synthesis of intracellular antigen and the expression of putative virulence factors by Renibacterium salmoninarum. EPC cultures infected with R. salmoninarum could be maintained for 7 weeks, during which the pathogen multiplied intracellularly. Immunohistochemical examination of infected cultures revealed the production of the p57 antigen, haemolysin and cytolysin. The intracellular nature of the infection was confirmed by transmission electron microscopic examination of EPC monolayers. A comparison of the relative virulence of bacterial cells cultured in EPC cells and on agar plates revealed that the former were markedly more virulent in challenge experiments with juvenile rainbow trout (Oncorhynchus mykiss Walbaum). The EPC cell culture model provided a system for the study of R. salmoninarum under more natural conditions than those achieved with plate culture techniques. PMID:9353936

  13. Anaerobic thermophilic culture system

    DOEpatents

    Ljungdahl, Lars G.; Wiegel, Jurgen K. W.

    1981-01-01

    A mixed culture system of the newly discovered microorganism Thermoanaerobacter ethanolicus ATCC31550 and the microorganism Clostridium thermocellum ATCC31549 is described. In a mixed nutrient culture medium that contains cellulose, these microorganisms have been coupled and cultivated to efficiently ferment cellulose to produce recoverable quantities of ethanol under anaerobic, thermophilic conditions.

  14. Evaluating 3 gas-delivery systems for culturing Campylobacter jejuni in a microaerophilic environment.

    PubMed

    Haines, M D; Eberle, K N; McDaniel, C D; Kiess, A S

    2011-10-01

    Campylobacter spp. require a microaerophilic environment (80% N(2), 10% CO(2), 5% H(2), and 5% O(2)) for growth. Since the late 1800s, several systems for creating and maintaining specific microbial atmospheres have been developed and applied. The objective of this study was to evaluate Campylobacter jejuni growth by means of 3 commonly used gas-delivery systems for generating a microaerophilic environment: automated, gas-generating sachet, and plastic storage bag. Pure culture C. jejuni cells were suspended in Brucella broth and spread onto campy cefex agar plates. For the automated gas-delivery system, plates were positioned in a Mart anaerobic jar and flushed with a microaerophilic gas mixture using an Anoxomat Mart II system (Mart Microbiology B. V., Netherlands). For the sachet samples, plates were placed in a Mart anaerobic jar and 3 Gaspak EZ campy sachets (Becton Dickinson and Company, Franklin Lakes, NJ) were activated to induce a microaerophilic gas environment. The plates placed in plastic storage bags were flushed with a microaerophilic gas mixture from a premixed tank. For all 3 systems, plates were placed in a low-temperature incubator at 42°C for 24 h. After 24 h, plates were removed from the incubator and colonies were counted. The entire experiment was repeated 5 times. Results indicated no significant difference in colony counts among the gas-delivery systems tested, but colonies grown under the sachet-generated environment were smaller than colonies in the other 2 methods. Smaller colonies could have resulted from the type of media used or the length of time the plates were incubated. In conclusion, all 3 gas-delivery methods were able to produce similar Campylobacter growth results. Initial and long-term costs of equipment, as well as laboratory space availability, may be influential when choosing a gas-delivery method for generating a microaerophilic environment. PMID:21934023

  15. Manuals of Cultural Systems

    NASA Astrophysics Data System (ADS)

    Ballonoff, Paul

    2014-10-01

    Ethnography often studies social networks including empirical descriptions of marriages and families. We initially concentrate on a special subset of networks which we call configurations. We show that descriptions of the possible outcomes of viable histories form a manual, and an orthoalgebra. We then study cases where family sizes vary, and show that this also forms a manual. In fact, it demonstrates adiabatic invariance, a property often associated with physical system conservation laws, and which here expresses conservation of the viability of a cultural system.

  16. MALDI-TOF mass spectrometry for early identification of bacteria grown in blood culture bottles.

    PubMed

    Zabbe, Jean-Benoît; Zanardo, Laura; Mégraud, Francis; Bessède, Emilie

    2015-08-01

    This note reports an interesting way to rapidly identify bacteria grown from blood culture bottles. Chocolate agar plates were inoculated with 1 drop of the positive blood bottle medium. After a 3-hour incubation, the growth veil was submitted to MALDI-TOF mass spectrometry: 77% of the bacteria present have been correctly identified. PMID:25940929

  17. Mass algal culture system

    DOEpatents

    Raymond, Lawrence P.

    1982-01-01

    An apparatus and process for the culture of algae in a liquid medium is disclosed. The medium circulates through an open trough and is exposed to an atmosphere which is temperature regulated. The nutrient content of the liquid medium is regulated to control the chemical composition growth and reproduction characteristics of the cultured algae. Before it is allowed to strike the medium, sunlight is passed through a filter to remove wavelengths which are not photosynthetically active. Heat energy can be recovered from the filter.

  18. Mass algal culture system

    DOEpatents

    Raymond, Lawrence P.

    1981-01-01

    An apparatus and process for the culture of algae in a liquid medium is disclosed. The medium circulates through an open trough and is exposed to an atmosphere which is temperature regulated. The nutrient content of the liquid medium is regulated to control the chemical composition growth and reproduction characteristics of the cultured algae. Before it is allowed to strike the medium, sunlight is passed through a filter to remove wavelengths which are not photosynthetically active. Heat energy can be recovered from the filter.

  19. High density cell culture system

    NASA Technical Reports Server (NTRS)

    Spaulding, Glenn F. (Inventor)

    1994-01-01

    An annular culture vessel for growing mammalian cells is constructed in a one piece integral and annular configuration with an open end which is closed by an endcap. The culture vessel is rotatable about a horizontal axis by use of conventional roller systems commonly used in culture laboratories. The end wall of the endcap has tapered access ports to frictionally and sealingly receive the ends of hypodermic syringes. The syringes permit the introduction of fresh nutrient and withdrawal of spent nutrients. The walls are made of conventional polymeric cell culture material and are subjected to neutron bombardment to form minute gas permeable perforations in the walls.

  20. Protein Chips for Detection of Salmonella spp. from Enrichment Culture

    PubMed Central

    Poltronieri, Palmiro; Cimaglia, Fabio; De Lorenzis, Enrico; Chiesa, Maurizio; Mezzolla, Valeria; Reca, Ida Barbara

    2016-01-01

    Food pathogens are the cause of foodborne epidemics, therefore there is a need to detect the pathogens in food productions rapidly. A pre-enrichment culture followed by selective agar plating are standard detection methods. Molecular methods such as qPCR have provided a first rapid protocol for detection of pathogens within 24 h of enrichment culture. Biosensors also may provide a rapid tool to individuate a source of Salmonella contamination at early times of pre-enrichment culture. Forty mL of Salmonella spp. enrichment culture were processed by immunoseparation using the Pathatrix, as in AFNOR validated qPCR protocols. The Salmonella biosensor combined with immunoseparation showed a limit of detection of 100 bacteria/40 mL, with a 400 fold increase to previous results. qPCR analysis requires processing of bead-bound bacteria with lysis buffer and DNA clean up, with a limit of detection of 2 cfu/50 μL. Finally, a protein chip was developed and tested in screening and identification of 5 common pathogen species, Salmonella spp., E. coli, S. aureus, Campylobacter spp. and Listeria spp. The protein chip, with high specificity in species identification, is proposed to be integrated into a Lab-on-Chip system, for rapid and reproducible screening of Salmonella spp. and other pathogen species contaminating food productions. PMID:27110786

  1. Protein Chips for Detection of Salmonella spp. from Enrichment Culture.

    PubMed

    Poltronieri, Palmiro; Cimaglia, Fabio; De Lorenzis, Enrico; Chiesa, Maurizio; Mezzolla, Valeria; Reca, Ida Barbara

    2016-01-01

    Food pathogens are the cause of foodborne epidemics, therefore there is a need to detect the pathogens in food productions rapidly. A pre-enrichment culture followed by selective agar plating are standard detection methods. Molecular methods such as qPCR have provided a first rapid protocol for detection of pathogens within 24 h of enrichment culture. Biosensors also may provide a rapid tool to individuate a source of Salmonella contamination at early times of pre-enrichment culture. Forty mL of Salmonella spp. enrichment culture were processed by immunoseparation using the Pathatrix, as in AFNOR validated qPCR protocols. The Salmonella biosensor combined with immunoseparation showed a limit of detection of 100 bacteria/40 mL, with a 400 fold increase to previous results. qPCR analysis requires processing of bead-bound bacteria with lysis buffer and DNA clean up, with a limit of detection of 2 cfu/50 μL. Finally, a protein chip was developed and tested in screening and identification of 5 common pathogen species, Salmonella spp., E. coli, S. aureus, Campylobacter spp. and Listeria spp. The protein chip, with high specificity in species identification, is proposed to be integrated into a Lab-on-Chip system, for rapid and reproducible screening of Salmonella spp. and other pathogen species contaminating food productions. PMID:27110786

  2. Economic Systems: A Modular Approach. Cultural Anthropology.

    ERIC Educational Resources Information Center

    Kassebaum, Peter

    Designed for use as supplementary instructional material in a cultural anthropology course, this learning module uses a systems approach to allow students to see the connections and similarities which most cultural groups share on the basis of the type of economic organization that they exhibit. The module begins with a general discussion of…

  3. Constructing a High Density Cell Culture System

    NASA Technical Reports Server (NTRS)

    Spaulding, Glenn F. (Inventor)

    1996-01-01

    An annular culture vessel for growing mammalian cells is constructed in a one piece integral and annular configuration with an open end which is closed by an endcap. The culture vessel is rotatable about a horizontal axis by use of conventional roller systems commonly used in culture laboratories. The end wall of the endcap has tapered access ports to frictionally and sealingly receive the ends of hypodermic syringes. The syringes permit the introduction of fresh nutrient and withdrawal of spent nutrients. The walls are made of conventional polymeric cell culture material and are subjected to neutron bombardment to form minute gas permeable perforations in the walls.

  4. Digital image quantification of siderophores on agar plates

    PubMed Central

    Andrews, Megan Y.; Santelli, Cara M.; Duckworth, Owen W.

    2016-01-01

    This article presents visual image data and detailed methodology for the use of a new method for quantifying the exudation of siderophores during fungal growth. The data include images showing time series for calibration, fungal exudation, and negative controls, as well as replication accuracy information. In addition, we provide detailed protocols for making CAS assay layer plates, the digital analysis protocol for determining area of color change, and discuss growth media that do and do not work with the layer plate method. The results of these data, their interpretation, and further discussion can be found in Andrews et al., 2016 [1]. PMID:26937467

  5. Digital image quantification of siderophores on agar plates.

    PubMed

    Andrews, Megan Y; Santelli, Cara M; Duckworth, Owen W

    2016-03-01

    This article presents visual image data and detailed methodology for the use of a new method for quantifying the exudation of siderophores during fungal growth. The data include images showing time series for calibration, fungal exudation, and negative controls, as well as replication accuracy information. In addition, we provide detailed protocols for making CAS assay layer plates, the digital analysis protocol for determining area of color change, and discuss growth media that do and do not work with the layer plate method. The results of these data, their interpretation, and further discussion can be found in Andrews et al., 2016 [1]. PMID:26937467

  6. Hyperspectral Imaging for Detecting Pathogens Grown on Agar Plates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This paper is concerned with the development of a hyperspectral imaging technique for detecting and identifying one of the most common foodborne pathogens, Campylobacter. Direct plating using agars is an effective tool for laboratory tests and analyses of microorganisms. The morphology (size, growth...

  7. A rapid method combining immunofluorescence and flow cytometry for improved understanding of competitive interactions between lactic acid bacteria (LAB) and methicillin-resistant S. aureus (MRSA) in mixed culture.

    PubMed

    Schellenberg, John; Smoragiewicz, Wanda; Karska-Wysocki, Barbara

    2006-04-01

    The increasing frequency of methicillin-resistant Staphylococcus aureus (MRSA) infections in hospital and community settings highlights the need for effective anti-MRSA agents that will not contribute to the growing problem of antibiotic resistance. Lactic acid bacteria (LAB) are known to exclude various pathogens through multiple mechanisms. In vitro models studying interactions of pathogens and LAB in mixed cultures use selective agar plates to quantify changes in target populations. We applied commercially available S. aureus-specific polyclonal antibodies conjugated with fluorescein isothiocyanate (FITC) for this purpose, producing a bright green signal that clearly differentiates S. aureus from LAB species when mixed cultures are analyzed by flow cytometry and fluorescent microscopy. Flow cytometry of mixed cultures revealed a much larger population of MRSA cells than was detectable using selective agar plates. To our knowledge, this is the first time immunofluorescent flow cytometry has been applied to the study of competitive exclusion in mixed bacterial populations over time. PMID:16154216

  8. Culturing Mouse Cardiac Valves in the Miniature Tissue Culture System.

    PubMed

    Kruithof, Boudewijn P T; Lieber, Samuel C; Kruithof-de Julio, Marianna; Gaussin, Vincian; Goumans, Marie José

    2015-01-01

    Heart valve disease is a major burden in the Western world and no effective treatment is available. This is mainly due to a lack of knowledge of the molecular, cellular and mechanical mechanisms underlying the maintenance and/or loss of the valvular structure. Current models used to study valvular biology include in vitro cultures of valvular endothelial and interstitial cells. Although, in vitro culturing models provide both cellular and molecular mechanisms, the mechanisms involved in the 3D-organization of the valve remain unclear. While in vivo models have provided insight into the molecular mechanisms underlying valvular development, insight into adult valvular biology is still elusive. In order to be able to study the regulation of the valvular 3D-organization on tissue, cellular and molecular levels, we have developed the Miniature Tissue Culture System. In this ex vivo flow model the mitral or the aortic valve is cultured in its natural position in the heart. The natural configuration and composition of the leaflet are maintained allowing the most natural response of the valvular cells to stimuli. The valves remain viable and are responsive to changing environmental conditions. This MTCS may provide advantages on studying questions including but not limited to, how does the 3D organization affect valvular biology, what factors affect 3D organization of the valve, and which network of signaling pathways regulates the 3D organization of the valve. PMID:26555276

  9. Culture of Helicobacter pylori from stool samples in children.

    PubMed

    Falsafi, Tahereh; Valizadeh, Nargess; Najafi, Mehri; Ehsani, Azadeh; Khani, Afsaneh; Landarani, Zahra; Falahi, Zahra

    2007-03-01

    We evaluated two protocols for isolation of Helicobacter pylori in stool from biopsied and nonbiopsied children. Twenty-three child patients whose presumptive positivity or negativity was diagnosed by endoscopy and a rapid urease test at site were used to compare biopsy-based tests with stool-based tests (H. pylori stool antigen test and stool culture). Their gastric activity and bacterial density were graded by the updated Sydney system. Biopsy and stool specimens were cultured on Campy-blood and Belo horizonte agar plates after enrichment in selective Campy-Thio medium. To compare two stool culture protocols, stools from 20 nonbiopsied children were tested by the HpSA test and cultured either as above or after treatment with cholestyramine. Grown colonies were screened by Gram staining, slide agglutination using anti-H. pylori monoclonal IgG; positive isolates were tested by biochemical tests and polymerase chain reaction for H. pylori-specific ureA gene. Coccoid H. pylori was isolated in stool samples from the biopsied patients whose bacterial density was two to four in histology. Their oxidase was slightly positive but became positive after two subcultures, while additional biochemical tests confirmed the isolation of H. pylori. Similar coccoid but oxidase positive H. pylori was isolated from three nonbiopsied children with the protocol of cholestyramine treatment only. The density of bacteria in the stomach may influence the recovery of H. pylori from stool; inactivation of bile with cholestyramine improves the yield in culture and favors isolation of an enhanced metabolic form of bacteria. PMID:17538651

  10. Evaluation of an Image Analysis Device (APAS) for Screening Urine Cultures.

    PubMed

    Glasson, John; Hill, Rhys; Summerford, Michael; Giglio, Steven

    2016-02-01

    While advancements have been made in some areas of pathology with diagnostic materials being screened using image analysis technologies, the reporting of cultures from agar plates remains a manual process. We compared the results for 2,163 urine cultures read by a reference panel of microbiologists, by the routine laboratory process, and by an automated plate reading system, APAS (LBT Innovations Ltd., South Australia). APAS detected colonies with a sensitivity of 99.1% and a specificity of 99.3% on blood agar, while on MacConkey agar, the colony detection sensitivity was 99.4% with a specificity of 99.3%. The device's ability to enumerate growth had an accuracy of 89.2%, and the morphological identification of colonies showed a high level of performance for the colony types typical of Escherichia coli and other enteric bacilli. On blood agar, lactose-fermenting colonies were morphologically identified with a sensitivity of 98.9%, while on MacConkey agar they were identified with a sensitivity of 99.2%. In this first clinical evaluation, APAS demonstrated high performance in the detection, enumeration, and colony classification of isolates compared with that for conventional plate-reading methods. The device found all cases reported by the laboratory and detected the most commonly encountered organisms found in urinary tract infections. PMID:26582838

  11. Evaluation of an Image Analysis Device (APAS) for Screening Urine Cultures

    PubMed Central

    Hill, Rhys; Summerford, Michael; Giglio, Steven

    2015-01-01

    While advancements have been made in some areas of pathology with diagnostic materials being screened using image analysis technologies, the reporting of cultures from agar plates remains a manual process. We compared the results for 2,163 urine cultures read by a reference panel of microbiologists, by the routine laboratory process, and by an automated plate reading system, APAS (LBT Innovations Ltd., South Australia). APAS detected colonies with a sensitivity of 99.1% and a specificity of 99.3% on blood agar, while on MacConkey agar, the colony detection sensitivity was 99.4% with a specificity of 99.3%. The device's ability to enumerate growth had an accuracy of 89.2%, and the morphological identification of colonies showed a high level of performance for the colony types typical of Escherichia coli and other enteric bacilli. On blood agar, lactose-fermenting colonies were morphologically identified with a sensitivity of 98.9%, while on MacConkey agar they were identified with a sensitivity of 99.2%. In this first clinical evaluation, APAS demonstrated high performance in the detection, enumeration, and colony classification of isolates compared with that for conventional plate-reading methods. The device found all cases reported by the laboratory and detected the most commonly encountered organisms found in urinary tract infections. PMID:26582838

  12. HBV culture and infectious systems.

    PubMed

    Hayes, C Nelson; Chayama, Kazuaki

    2016-07-01

    While an effective vaccine against hepatitis B virus (HBV) has long been available, chronic HBV infection remains a severe global public health concern. Current treatment options have limited effectiveness, and long-term therapy is required to suppress HBV replication; however, complete elimination of the virus is rare. The lack of suitable animal models and infection systems has hindered efforts to unravel the HBV life cycle, particularly the early events in HBV entry, which appear to be highly species- and tissue-specific. Human primary hepatocytes remain the gold standard for HBV replication studies but are limited by availability and variability. While the HepaRG cell line is permissive for HBV replication, other hepatoma cell lines such as HepG2 do not support HBV replication. The recent discovery of sodium taurocholate transporting peptide (NTCP) as a primary receptor for HBV binding has led to the development of replication-competent cell lines such as HepG2-NTCP. Human hepatocytes grown in chimeric mice have provided another approach that allows primary human hepatocytes to be used while overcoming many of their limitations. Although the difficulty in developing HBV infection systems has hindered development of effective treatments, the variability and limited replication efficiency among cell lines point to additional liver-specific factors involved in HBV infection. It is hoped that HBV infection studies will lead to novel drug targets and therapeutic options for the treatment of chronic HBV infection. PMID:26935052

  13. Label-free isolation and deposition of single bacterial cells from heterogeneous samples for clonal culturing.

    PubMed

    Riba, J; Gleichmann, T; Zimmermann, S; Zengerle, R; Koltay, P

    2016-01-01

    The isolation and analysis of single prokaryotic cells down to 1 μm and less in size poses a special challenge and requires micro-engineered devices to handle volumes in the picoliter to nanoliter range. Here, an advanced Single-Cell Printer (SCP) was applied for automated and label-free isolation and deposition of bacterial cells encapsulated in 35 pl droplets by inkjet-like printing. To achieve this, dispenser chips to generate micro droplets have been fabricated with nozzles 20 μm in size. Further, the magnification of the optical system used for cell detection was increased. Redesign of the optical path allows for collision-free addressing of any flat substrate since no compartment protrudes below the nozzle of the dispenser chip anymore. The improved system allows for deterministic isolation of individual bacterial cells. A single-cell printing efficiency of 93% was obtained as shown by printing fluorescent labeled E. coli. A 96-well plate filled with growth medium is inoculated with single bacteria cells on average within about 8 min. Finally, individual bacterial cells from a heterogeneous sample of E. coli and E. faecalis were isolated for clonal culturing directly on agar plates in user-defined array geometry. PMID:27596612

  14. Label-free isolation and deposition of single bacterial cells from heterogeneous samples for clonal culturing

    PubMed Central

    Riba, J.; Gleichmann, T.; Zimmermann, S.; Zengerle, R.; Koltay, P.

    2016-01-01

    The isolation and analysis of single prokaryotic cells down to 1 μm and less in size poses a special challenge and requires micro-engineered devices to handle volumes in the picoliter to nanoliter range. Here, an advanced Single-Cell Printer (SCP) was applied for automated and label-free isolation and deposition of bacterial cells encapsulated in 35 pl droplets by inkjet-like printing. To achieve this, dispenser chips to generate micro droplets have been fabricated with nozzles 20 μm in size. Further, the magnification of the optical system used for cell detection was increased. Redesign of the optical path allows for collision-free addressing of any flat substrate since no compartment protrudes below the nozzle of the dispenser chip anymore. The improved system allows for deterministic isolation of individual bacterial cells. A single-cell printing efficiency of 93% was obtained as shown by printing fluorescent labeled E. coli. A 96-well plate filled with growth medium is inoculated with single bacteria cells on average within about 8 min. Finally, individual bacterial cells from a heterogeneous sample of E. coli and E. faecalis were isolated for clonal culturing directly on agar plates in user-defined array geometry. PMID:27596612

  15. Culture systems for hepatitis E virus.

    PubMed

    Okamoto, Hiroaki

    2013-02-01

    The lack of an efficient cell culture system for hepatitis E virus (HEV) has greatly hampered detailed analyses of this virus. The first efficient cell culture systems for HEV that were developed were capable of secreting infectious HEV progenies in high titers into culture media, using PLC/PRF/5 cells derived from human hepatocellular carcinoma and A549 cells derived from human lung cancer as host cells. The success achieved with the original genotype 3 JE03-1760F strain has now been extended to various HEV strains in fecal and serum samples obtained from hepatitis E patients and to HEV strains in fecal and serum samples and liver tissues obtained from pigs and wild boar across species barriers. In addition, infectious HEV cDNA clones of the wild-type JE03-1760F strain and its variants have been engineered. Cell culture-generated HEV particles and those in circulating blood were found to be associated with lipids and open reading frame 3 (ORF3) protein, thereby likely contributing to the assembly and release of HEV from infected cells both in vivo and in vitro. The ORF3 protein interacts with the tumor susceptibility gene 101, a critical cellular protein required for the budding of enveloped viruses, through the Pro, Ser, Ala, and Pro (PSAP) motif in infected cells; ORF3 is co-localized with multivesicular bodies (MVBs) in the cytoplasm of infected cells, thus suggesting that HEV requires the MVB pathway for the egress of virus particles. This article reviews the development of efficient cell culture systems for a wide variety of infectious HEV strains obtained from humans, pigs, and wild boar, and also provides details of a new model for virion egress. PMID:23104469

  16. The use of glycylhistidyllysine in culture systems.

    PubMed

    Pickart, L

    1981-06-01

    Glycylhistidyllysine (GHL), a tripeptide isolated from plasma, has been shown to alter the growth rate of many cell types and organisms in culture systems. The tripeptide is optimally active at concentrations between 10 and 200 ng/ml. Some of the more interesting uses of GHL are highlighted in this paper. Present information suggests that GHL functions as a transporter of transition metals, in particular copper, to the cell surface for uptake into the cell. PMID:7021400

  17. Dynamic cell culture system (7-IML-1)

    NASA Technical Reports Server (NTRS)

    Cogoli, Augusto

    1992-01-01

    This experiment is one of the Biorack experiments being flown on the International Microgravity Laboratory 1 (MIL-1) mission as part of an investigation studying cell proliferation and performance in space. One of the objectives of this investigation is to assess the potential benefits of bioprocessing in space with the ultimate goal of developing a bioreactor for continuous cell cultures in space. This experiment will test the operation of an automated culture chamber that was designed for use in a Bioreactor in space. The device to be tested is called the Dynamic Cell Culture System (DCCS). It is a simple device in which media are renewed or chemicals are injected automatically, by means of osmotic pumps. This experiment uses four Type I/O experiment containers. One DCCS unit, which contains a culture chamber with renewal of medium and a second chamber without a medium supply fits in each container. Two DCCS units are maintained under zero gravity conditions during the on-orbit period. The other two units are maintained under 1 gh conditions in a 1 g centrifuge. The schedule for incubator transfer is given.

  18. Comparison of Identification Systems for Classification of Bacteria Isolated from Water and Endolithic Habitats within the Deep Subsurface

    PubMed Central

    Amy, P. S.; Haldeman, D. L.; Ringelberg, D.; Hall, D. H.; Russell, C.

    1992-01-01

    One water and three rock samples were taken from a mined tunnel system, U12n, in Rainier Mesa at the Nevada Test Site. Endolithic microorganisms were cultured from ashfall tuff, which was crushed and made into slurries with a formulation of artificial pore water, on R2A agar plates. Microbial counts ranged from 102 to 104 viable cells per g (dry weight) of rock sampled. The cultured water sample yielded 102 viable cells per ml. Many of the isolates were very small (<1 μm) when viewed in the rock matrix and remained small even when cultured. Most were gram-negative rods. Individual isolates were profiled by API-NFT strip number, antibiotic and metal resistance patterns, and colony and cellular morphologies. Three identification systems, API-NFT strips, BIOLOG, and MIDI, were compared. Each system identified only a small percentage of the total isolates, and in only seven cases were the isolates identified the same way by more than one system. The same genus was identified in three of these cases, but different species were indicated. The genus Pseudomonas was the most commonly identified. The isolate profiles and the three identification systems demonstrated that water isolates were considerably different from endolithic isolates. PMID:16348791

  19. Thin agar film for enhanced fungal growth and microscopic viewing in a new sealable fungal culture case.

    PubMed

    Hill, D R

    1996-09-01

    This project was undertaken to find ways to enhance fungus colony maturation, to make viewing of fungal cultures easier, and to reduce disruption of the fungal structures to be observed for identification. Accordingly, a technique using a thin (0.2-mm) agar film that avoids problems inherent in traditional methods of fungal culture and identification was developed. In addition, to accommodate the 0.2-mm layer of agar film and a contiguous thicker 4-mm section of agar, a sealable fungal culture case that fits within microscope stage calipers and under the objective lenses was invented. The growth and identification of 28 organisms were evaluated in the sealable fungal culture cases and on double-pour agar plates by using potato dextrose agar in both. Compared with results obtained with the double-pour agar plates (rated as "good"), fungal growth and identification with the sealable fungal culture case were superior (rated as "excellent") (P < 0.05, chi-square test). The thin agar film limits excessive mycelial growth, while it often promotes complete sporulation or other forms of maturation of the fungal colony. More importantly, the thin agar film allows direct microscopic viewing of the developing fungal colonies. The portion of the sealable fungal culture case with the 4-mm layer of agar can be used for evaluation of colony pigment and texture. In conclusion, this new sealable fungal culture case allows direct viewing and earlier fungal species identification with greater intrinsic safety. PMID:8862573

  20. Correlation between mastitis occurrence and the count of microorganisms in bulk raw milk of bovine dairy herds in four selective culture media.

    PubMed

    Souto, Luís I M; Minagawa, Clarice Y; Telles, Evelise O; Garbuglio, Márcio A; Amaku, Marcos; Melville, Priscilla A; Dias, Ricardo A; Sakata, Sonia T; Benites, Nilson R

    2010-02-01

    Milk is the normal secretion of the mammary gland, practically free of colostrum and obtained by the complete milking of one or more healthy animals. Mastitis is an inflammatory process of the mammary gland and it may cause alterations in the milk. The present work aimed to verify whether it is possible, by means of the counts of microorganism in the bulk raw milk in four selective culture media, to establish a correlation with the occurrence of mastitis and therefore, to monitor this disease in bovine dairy herds. The following selective culture media were used: KF Streptococcus Agar, Edwards Agar, Baird-Parker Agar, Blood Agar plus potassium tellurite. Spearman's correlation coefficient was calculated in order to compare the occurrence of mastitis (percentage) in each herd with respective selective culture media counts of microorganisms in bulk raw milk. Thirty-six possibilities were analysed (Tamis and CMT-positive rates were compared with the log-transformed count in four selective culture media) and there was a negative correlation between Tamis 3 and the Baird-Parker Agar plate count. The total results of microbiological tests showed that there were three correlations of the counts in selective culture media. Fifty-two possibilities were analysed and there was a negative correlation between no-bacterial-growth mastitis rates and log10 of KF Streptoccocus Agar plate count and there were two positive correlations between coagulase-positive staphylococci and log10 of Baird-Parker Agar plate count and Blood Agar plus potassium tellurite plate count. PMID:19939321

  1. Cultural systems for growing potatoes in space

    NASA Technical Reports Server (NTRS)

    Tibbitts, T.; Bula, R.; Corey, R.; Morrow, R.

    1988-01-01

    Higher plants are being evaluated for life support to provide needed food, oxygen and water as well as removal of carbon dioxide from the atmosphere. The successful utilization of plants in space will require the development of not only highly productive growing systems but also highly efficient bioregenerative systems. It will be necessary to recycle all inedible plant parts and all human wastes so that the entire complement of elemental compounds can be reused. Potatoes have been proposed as one of the desirable crops because they are 1) extremely productive, yielding more than 100 metric tons per hectare from field plantings, 2) the edible tubers are high in digestible starch (70%) and protein (10%) on a dry weight basis, 3) up to 80% of the total plant production is in tubers and thus edible, 4) the plants are easily propagated either from tubers or from tissue culture plantlets, 5) the tubers can be utilized with a minimum of processing, and 6) potatoes can be prepared in a variety of different forms for the human diet (Tibbitts et al., 1982). However potatoes have a growth pattern that complicates the development of growing the plants in controlled systems. Tubers are borne on underground stems that are botanically termed 'rhizomes', but in common usage termed 'stolons'. The stolons must be maintained in a dark, moist area with sufficient provision for enlargement of tubers. Stems rapidly terminate in flowers forcing extensive branching and spreading of plants so that individual plants will cover 0.2 m2 or more area. Thus the growing system must be developed to provide an area that is darkened for tuber and root growth and of sufficient size for plant spread. A system developed for growing potatoes, or any plants, in space will have certain requirements that must be met to make them a useful part of a life support system. The system must 1) be constructed of materials, and involve media, that can be reused for many successive cycles of plant growth, 2

  2. Cultured Inquiry

    ERIC Educational Resources Information Center

    Scheppler, Judith A.; Sethakorn, Nan; Styer, Susan

    2003-01-01

    The Kirby-Bauer assay, also called the disc diffusion assay, is a standard procedure used in clinical laboratories to test the susceptibility of patients' bacterial isolates to antibiotics. In the assay, the bacteria are swabbed onto an agar plate, and paper discs impregnated with antibiotics are placed on the agar. The antibiotic diffuses from…

  3. Oxygenation of intensive cell-culture system.

    PubMed

    Emery, A N; Jan, D C; al-Rubeai, M

    1995-11-01

    The abilities of various methods of oxygenation to meet the demands of high-cell-density culture were investigated using a spin filter perfusion system in a bench-top bioreactor. Oxygen demand at high cell density could not be met by sparging with air inside a spin filter (oxygen transfer values in this condition were comparable with those for surface aeration). Sparging with air outside a spin filter gave adequate oxygen transfer for the support of cell concentrations above 10(7) ml-1 in fully aerobic conditions but the addition of antifoam to control foaming caused blockage of the spinfilter mesh. Bubble-free aeration through immersed silicone tubing with pure oxygen gave similar oxygen transfer rates to that of sparging with air but without the problems of bubble damage and fouling of the spin filter. A supra-optimal level of dissolved oxygen (478% air saturation) inhibited cell growth. However, cells could recover from this stress and reach high density after reduction of the dissolved oxygen level to 50% air saturation. PMID:8590652

  4. Functional Systems and Culturally-Determined Cognitive Differences.

    ERIC Educational Resources Information Center

    Wiseman, Richard L.

    Noting that one means of better understanding the nature of cultural differences is to elucidate the cognitive differences between members of differing cultures, this paper examines Alexander Luria's sociohistorical theory of functional cognitive systems. The paper first describes Luria's notion of functional systems, the crux of which postulates…

  5. Towards a Culturally Competent System of Care. Volume II: Programs Which Utilize Culturally Competent Principles.

    ERIC Educational Resources Information Center

    Isaacs, Mareasa R.; Benjamin, Marva P.

    This monograph was developed to assist states and communities in planning, designing, and implementing culturally competent systems of care for children with serious emotional disturbances. It highlights the culturally competent aspects of 11 programs serving people of color. Each program targets at least one of the four major ethnic minority…

  6. Evaluation of Verigene Blood Culture Test Systems for Rapid Identification of Positive Blood Cultures

    PubMed Central

    Kim, Jae-Seok; Kang, Go-Eun; Kim, Han-Sung; Song, Wonkeun; Lee, Kyu Man

    2016-01-01

    The performance of molecular tests using the Verigene Gram-Positive and Gram-Negative Blood Culture nucleic acid tests (BC-GP and BC-GN, resp.; Naosphere, Northbrook, IL, USA) was evaluated for the identification of microorganisms detected from blood cultures. Ninety-nine blood cultures containing Gram-positive bacteria and 150 containing Gram-negative bacteria were analyzed using the BC-GP and BC-GN assays, respectively. Blood cultures were performed using the Bactec blood culture system (BD Diagnostic Systems, Franklin Lakes, NJ, USA) and conventional identification and antibiotic-susceptibility tests were performed using a MicroScan system (Siemens, West Sacramento, CA, USA). When a single strain of bacteria was isolated from the blood culture, Verigene assays correctly identified 97.9% (94/96) of Gram-positive bacteria and 93.8% (137/146) of Gram-negative bacteria. Resistance genes mecA and vanA were correctly detected by the BC-GP assay, while the extended-spectrum β-lactamase CTX-M and the carbapenemase OXA resistance gene were detected from 30 cases cultures by the BC-GN assay. The BC-GP and BC-GN assays showed high agreement with conventional identification and susceptibility tests. These tests are useful for rapid identification of microorganisms and the detection of clinically important resistance genes from positive Bactec blood cultures. PMID:26904669

  7. Anaerobic bag culture method.

    PubMed

    Rosenblatt, J E; Stewart, P R

    1975-06-01

    In a new method of anaerobic culture, a transparent, gas-impermeable bag is used and the anaerobic environment is established with copper sulfate-saturated steel wool. An Alka-Seltzer tablet generates carbon dioxide. The agar plate surface can be inspected through the bag at any time without interrupting the anaerobic atmosphere or disturbing other specimens. Methylene blue indicator strips are completely reduced by 4 h after the bag is set up and have remained reduced for as long as 3 weeks. Growth of 16 different stock culture anaerobes was generally equivalent by the bag and GasPak jar methods. Yield and growth of anaerobic isolates also were equivalent with 7 of 10 clinical specimens; from the other 3 specimens, 13 isolates were recovered, 5 by both the bag and jar methods and the rest by one method or the other. No consistent differences were found between the anaerobic bag and GasPak jar methods in the yield of anaerobes from clinical specimens. Early growth (24 h of incubation) of anaerobes from one specimen was detected with the bag method. PMID:1100671

  8. Anaerobic bag culture method.

    PubMed Central

    Rosenblatt, J E; Stewart, P R

    1975-01-01

    In a new method of anaerobic culture, a transparent, gas-impermeable bag is used and the anaerobic environment is established with copper sulfate-saturated steel wool. An Alka-Seltzer tablet generates carbon dioxide. The agar plate surface can be inspected through the bag at any time without interrupting the anaerobic atmosphere or disturbing other specimens. Methylene blue indicator strips are completely reduced by 4 h after the bag is set up and have remained reduced for as long as 3 weeks. Growth of 16 different stock culture anaerobes was generally equivalent by the bag and GasPak jar methods. Yield and growth of anaerobic isolates also were equivalent with 7 of 10 clinical specimens; from the other 3 specimens, 13 isolates were recovered, 5 by both the bag and jar methods and the rest by one method or the other. No consistent differences were found between the anaerobic bag and GasPak jar methods in the yield of anaerobes from clinical specimens. Early growth (24 h of incubation) of anaerobes from one specimen was detected with the bag method. Images PMID:1100671

  9. Plasticity of the Arabidopsis root system under nutrient deficiencies.

    PubMed

    Gruber, Benjamin D; Giehl, Ricardo F H; Friedel, Swetlana; von Wirén, Nicolaus

    2013-09-01

    Plant roots show a particularly high variation in their morphological response to different nutrient deficiencies. Although such changes often determine the nutrient efficiency or stress tolerance of plants, it is surprising that a comprehensive and comparative analysis of root morphological responses to different nutrient deficiencies has not yet been conducted. Since one reason for this is an inherent difficulty in obtaining nutrient-deficient conditions in agar culture, we first identified conditions appropriate for producing nutrient-deficient plants on agar plates. Based on a careful selection of agar specifically for each nutrient being considered, we grew Arabidopsis (Arabidopsis thaliana) plants at four levels of deficiency for 12 nutrients and quantified seven root traits. In combination with measurements of biomass and elemental concentrations, we observed that the nutritional status and type of nutrient determined the extent and type of changes in root system architecture (RSA). The independent regulation of individual root traits further pointed to a differential sensitivity of root tissues to nutrient limitations. To capture the variation in RSA under different nutrient supplies, we used principal component analysis and developed a root plasticity chart representing the overall modulations in RSA under a given treatment. This systematic comparison of RSA responses to nutrient deficiencies provides a comprehensive view of the overall changes in root plasticity induced by the deficiency of single nutrients and provides a solid basis for the identification of nutrient-sensitive steps in the root developmental program. PMID:23852440

  10. Biona-C Cell Culture pH Monitoring System

    NASA Technical Reports Server (NTRS)

    Friedericks, C.

    1999-01-01

    Sensors 2000! is developing a system to demonstrate the ability to perform accurate, real-time measurements of pH and CO2 in a cell culture media in Space. The BIONA-C Cell Culture pH Monitoring System consists of S2K! developed ion selective sensors and control electronics integrated with the fluidics of a cell culture system. The integrated system comprises a "rail" in the Cell Culture Module (CCM) of WRAIR (Space Biosciences of Walter Read Army Institute of Research). The CCM is a Space Shuttle mid-deck locker experiment payload. The BIONA-C is displayed along with associated graphics and text explanations. The presentation will stimulate interest in development of sensor technology for real-time cell culture measurements. The transfer of this technology to other applications will also be of interest. Additional information is contained in the original document.

  11. Cultural Development through Human Resource Systems Integration.

    ERIC Educational Resources Information Center

    Albert, Michael

    1985-01-01

    Discusses the framework for developing a cultural human resources management (HRM) perspective. Central to this framework is modifying HRM programs to reinforce the organization's preferred practices. Modification occurs through selection, orientation, training and development, performance appraisal, career development, and compensation and…

  12. Sustaining organizational culture change in health systems.

    PubMed

    Willis, Cameron David; Saul, Jessie; Bevan, Helen; Scheirer, Mary Ann; Best, Allan; Greenhalgh, Trisha; Mannion, Russell; Cornelissen, Evelyn; Howland, David; Jenkins, Emily; Bitz, Jennifer

    2016-03-21

    Purpose - The questions addressed by this review are: first, what are the guiding principles underlying efforts to stimulate sustained cultural change; second, what are the mechanisms by which these principles operate; and, finally, what are the contextual factors that influence the likelihood of these principles being effective? The paper aims to discuss these issues. Design/methodology/approach - The authors conducted a literature review informed by rapid realist review methodology that examined how interventions interact with contexts and mechanisms to influence the sustainability of cultural change. Reference and expert panelists assisted in refining the research questions, systematically searching published and grey literature, and helping to identify interactions between interventions, mechanisms and contexts. Findings - Six guiding principles were identified: align vision and action; make incremental changes within a comprehensive transformation strategy; foster distributed leadership; promote staff engagement; create collaborative relationships; and continuously assess and learn from change. These principles interact with contextual elements such as local power distributions, pre-existing values and beliefs and readiness to engage. Mechanisms influencing how these principles sustain cultural change include activation of a shared sense of urgency and fostering flexible levels of engagement. Practical implications - The principles identified in this review, along with the contexts and mechanisms that influence their effectiveness, are useful domains for policy and practice leaders to explore when grappling with cultural change. These principles are sufficiently broad to allow local flexibilities in adoption and application. Originality/value - This is the first study to adopt a realist approach for understanding how changes in organizational culture may be sustained. Through doing so, this review highlights the broad principles by which organizational action

  13. Growing B Lymphocytes in a Three-Dimensional Culture System

    NASA Technical Reports Server (NTRS)

    Wu, J. H. David; Bottaro, Andrea

    2010-01-01

    A three-dimensional (3D) culture system for growing long-lived B lymphocytes has been invented. The capabilities afforded by the system can be expected to expand the range of options for immunological research and related activities, including testing of immunogenicity of vaccine candidates in vitro, generation of human monoclonal antibodies, and immunotherapy. Mature lymphocytes, which are the effectors of adaptive immune responses in vertebrates, are extremely susceptible to apoptotic death, and depend on continuous reception of survival-inducing stimulation (in the forms of cytokines, cell-to-cell contacts, and antigen receptor signaling) from the microenvironment. For this reason, efforts to develop systems for long-term culture of functional, non-transformed and non-activated mature lymphocytes have been unsuccessful until now. The bone-marrow microenvironment supports the growth and differentiation of many hematopoietic lineages, in addition to B-lymphocytes. Primary bone-marrow cell cultures designed to promote the development of specific cell types in vitro are highly desirable experimental systems, amenable to manipulation under controlled conditions. However, the dynamic and complex network of stromal cells and insoluble matrix proteins is disrupted in prior plate- and flask-based culture systems, wherein the microenvironments have a predominantly two-dimensional (2D) character. In 2D bone-marrow cultures, normal B-lymphoid cells become progressively skewed toward precursor B-cell populations that do not retain a normal immunophenotype, and such mature B-lymphocytes as those harvested from the spleen or lymph nodes do not survive beyond several days ex vivo in the absence of mitogenic stimulation. The present 3D culture system is a bioreactor that contains highly porous artificial scaffolding that supports the long-term culture of bone marrow, spleen, and lymph-node samples. In this system, unlike in 2D culture systems, B-cell subpopulations developing

  14. Increased exosome production from tumour cell cultures using the Integra CELLine Culture System.

    PubMed

    Mitchell, J Paul; Court, Jacqueline; Mason, Malcolm David; Tabi, Zsuzsanna; Clayton, Aled

    2008-06-01

    Exosomes are nanometer-sized vesicles, secreted from most cell types, with documented immune-modulatory functions. Exosomes can be purified from cultured cells but to do so effectively, requires maintenance of cells at high density in order to obtain sufficient accumulation of exosomes in the culture medium, prior to purification. Whilst high density cultures can be achieved with cells in suspension, this remains difficult with adherent cells, resulting in low quantity of exosomes for subsequent study. We have used the Integra CELLine culture system, originally designed for hybridoma cultures, to achieve a significant increase in obtainable exosomes from adherent and non-adherent tumour cells. Traditional cultures of mesothelioma cells (cultured in 75 cm(2) flasks) gave an average yield of 0.78 microg+/-0.14 microg exosome/ml of conditioned medium. The CELLine Adhere 1000 (CLAD1000) flask, housing the same cell line, increased exosome yield approximately 12 fold to 10.06 microg+/-0.97 microg/ml. The morphology, phenotype and immune function of these exosomes were compared, and found to be identical in all respects. Similarly an 8 fold increase in exosome production was obtained from NKL cells (a suspension cell line) using a CELLine 1000 (CL1000) flask. The CELLine system also incurred ~5.5 fold less cost and reduced labour for cell maintenance. This simple culture system is a cost effective, useful method for significantly increasing the quantity of exosomes available from cultured cells, without detrimental effects. This tool should prove advantageous in future studies of exosome-immune modulation in cancer and other settings. PMID:18423480

  15. a Cultural Landscape Information System Developed with Open Source Tools

    NASA Astrophysics Data System (ADS)

    Chudyk, C.; Müller, H.; Uhler, M.; Würriehausen, F.

    2013-07-01

    Since 2010, the state of Rhineland-Palatinate in Germany has developed a cultural landscape information system as a process to secure and further enrich aggregate data about its cultural assets. In an open dialogue between governing authorities and citizens, the intention of the project is an active cooperation of public and private actors. A cultural landscape information system called KuLIS was designed as a web platform, combining semantic wiki software with a geographic information system. Based on data sets from public administrations, the information about cultural assets can be extended and enhanced by interested participants. The developed infrastructure facilitates local information accumulation through a crowdsourcing approach. This capability offers new possibilities for e-governance and open data developments. The collaborative approach allows governing authorities to manage and supervise official data, while public participation enables affordable information acquisition. Gathered cultural heritage information can provide incentives for touristic valorisation of communities or concepts for strengthening regional identification. It can also influence political decisions in defining significant cultural regions worth of protecting from industrial influences. The presented cultural landscape information allows citizens to influence the statewide development of cultural landscapes in a democratic way.

  16. Miniature Bioreactor System for Long-Term Cell Culture

    NASA Technical Reports Server (NTRS)

    Gonda, Steve R.; Kleis, Stanley J.; Geffert, Sandara K.

    2010-01-01

    A prototype miniature bioreactor system is designed to serve as a laboratory benchtop cell-culturing system that minimizes the need for relatively expensive equipment and reagents and can be operated under computer control, thereby reducing the time and effort required of human investigators and reducing uncertainty in results. The system includes a bioreactor, a fluid-handling subsystem, a chamber wherein the bioreactor is maintained in a controlled atmosphere at a controlled temperature, and associated control subsystems. The system can be used to culture both anchorage-dependent and suspension cells, which can be either prokaryotic or eukaryotic. Cells can be cultured for extended periods of time in this system, and samples of cells can be extracted and analyzed at specified intervals. By integrating this system with one or more microanalytical instrument(s), one can construct a complete automated analytical system that can be tailored to perform one or more of a large variety of assays.

  17. Strategy to obtain axenic cultures from field-collected samples of the cyanobacterium Phormidium animalis.

    PubMed

    Vázquez-Martínez, Guadalupe; Rodriguez, Mario H; Hernández-Hernández, Fidel; Ibarra, Jorge E

    2004-04-01

    An efficient strategy, based on a combination of procedures, was developed to obtain axenic cultures from field-collected samples of the cyanobacterium Phormidium animalis. Samples were initially cultured in solid ASN-10 medium, and a crude separation of major contaminants from P. animalis filaments was achieved by washing in a series of centrifugations and resuspensions in liquid medium. Then, manageable filament fragments were obtained by probe sonication. Fragmentation was followed by forceful washing, using vacuum-driven filtration through an 8-microm pore size membrane and an excess of water. Washed fragments were cultured and treated with a sequential exposure to four different antibiotics. Finally, axenic cultures were obtained from serial dilutions of treated fragments. Monitoring under microscope examination and by inoculation in Luria-Bertani (LB) agar plates indicated either axenicity or the degree of contamination throughout the strategy. PMID:15003694

  18. Association of heterotrophic bacteria with aggregated Arthrospira platensis exopolysaccharides: implications in the induction of axenic cultures.

    PubMed

    Shiraishi, Hideaki

    2015-01-01

    Inducing an axenic culture of the edible cyanobacterium Arthrospira (Spirulina) platensis using differential filtration alone is never successful; thus, it has been thought that, in non-axenic cultures, a portion of contaminating bacteria is strongly associated with Arthrospira cells. However, examination of the behavior of these bacteria during filtration revealed that they were not associated with Arthrospira cells but with aggregates of exopolysaccharides present in the medium away from the Arthrospira cells. Based on this finding, a rapid and reliable method for preparing axenic trichomes of A. platensis was established. After verifying the axenicity of the resulting trichomes on enriched agar plates, they were individually transferred to fresh sterile medium using a handmade tool, a microtrowel, to produce axenic cultures. With this technique, axenic cultures of various A. platensis strains were successfully produced. The technique described in this study is potentially applicable to a wider range of filamentous cyanobacteria. PMID:25333502

  19. Differentiation of mammalian skeletal muscle cells cultured on microcarrier beads in a rotating cell culture system

    NASA Technical Reports Server (NTRS)

    Torgan, C. E.; Burge, S. S.; Collinsworth, A. M.; Truskey, G. A.; Kraus, W. E.

    2000-01-01

    The growth and repair of adult skeletal muscle are due in part to activation of muscle precursor cells, commonly known as satellite cells or myoblasts. These cells are responsive to a variety of environmental cues, including mechanical stimuli. The overall goal of the research is to examine the role of mechanical signalling mechanisms in muscle growth and plasticity through utilisation of cell culture systems where other potential signalling pathways (i.e. chemical and electrical stimuli) are controlled. To explore the effects of decreased mechanical loading on muscle differentiation, mammalian myoblasts are cultured in a bioreactor (rotating cell culture system), a model that has been utilised to simulate microgravity. C2C12 murine myoblasts are cultured on microcarrier beads in a bioreactor and followed throughout differentiation as they form a network of multinucleated myotubes. In comparison with three-dimensional control cultures that consist of myoblasts cultured on microcarrier beads in teflon bags, myoblasts cultured in the bioreactor exhibit an attenuation in differentiation. This is demonstrated by reduced immunohistochemical staining for myogenin and alpha-actinin. Western analysis shows a decrease, in bioreactor cultures compared with control cultures, in levels of the contractile proteins myosin (47% decrease, p < 0.01) and tropomyosin (63% decrease, p < 0.01). Hydrodynamic measurements indicate that the decrease in differentiation may be due, at least in part, to fluid stresses acting on the myotubes. In addition, constraints on aggregate size imposed by the action of fluid forces in the bioreactor affect differentiation. These results may have implications for muscle growth and repair during spaceflight.

  20. Optimization of Methanotrophic Growth and Production of Poly(3-Hydroxybutyrate) in a High-Throughput Microbioreactor System.

    PubMed

    Sundstrom, Eric R; Criddle, Craig S

    2015-07-01

    Production of poly(3-hydroxybutyrate) (P3HB) from methane has economic and environmental advantages over production by agricultural feedstock. Identification of high-productivity strains and optimal growth conditions is critical to efficient conversion of methane to polymer. Current culture conditions, including serum bottles, shake flasks, and agar plates, are labor-intensive and therefore insufficient for systematic screening and isolation. Gas chromatography, the standard method for analysis of P3HB content in bacterial biomass, is also incompatible with high-throughput screening. Growth in aerated microtiter plates coupled with a 96-well Nile red flow-cytometric assay creates an integrated microbioreactor system for high-throughput growth and analysis of P3HB-producing methanotrophic cultures, eliminating the need for individual manipulation of experimental replicates. This system was tested in practice to conduct medium optimization for P3HB production in pure cultures of Methylocystis parvus OBBP. Optimization gave insight into unexpected interactions: for example, low calcium concentrations significantly enhanced P3HB production under nitrogen-limited conditions. Optimization of calcium and copper concentrations in the growth medium increased final P3HB content from 18.1% to 49.4% and P3HB concentration from 0.69 g/liter to 3.43 g/liter while reducing doubling time from 10.6 h to 8.6 h. The ability to culture and analyze thousands of replicates with high mass transfer in completely mixed culture promises to streamline medium optimization and allow the detection and isolation of highly productive strains. Applications for this system are numerous, encompassing analysis of biofuels and other lipid inclusions, as well as analysis of heterotrophic and photosynthetic systems. PMID:25956771

  1. Optimization of Methanotrophic Growth and Production of Poly(3-Hydroxybutyrate) in a High-Throughput Microbioreactor System

    PubMed Central

    Criddle, Craig S.

    2015-01-01

    Production of poly(3-hydroxybutyrate) (P3HB) from methane has economic and environmental advantages over production by agricultural feedstock. Identification of high-productivity strains and optimal growth conditions is critical to efficient conversion of methane to polymer. Current culture conditions, including serum bottles, shake flasks, and agar plates, are labor-intensive and therefore insufficient for systematic screening and isolation. Gas chromatography, the standard method for analysis of P3HB content in bacterial biomass, is also incompatible with high-throughput screening. Growth in aerated microtiter plates coupled with a 96-well Nile red flow-cytometric assay creates an integrated microbioreactor system for high-throughput growth and analysis of P3HB-producing methanotrophic cultures, eliminating the need for individual manipulation of experimental replicates. This system was tested in practice to conduct medium optimization for P3HB production in pure cultures of Methylocystis parvus OBBP. Optimization gave insight into unexpected interactions: for example, low calcium concentrations significantly enhanced P3HB production under nitrogen-limited conditions. Optimization of calcium and copper concentrations in the growth medium increased final P3HB content from 18.1% to 49.4% and P3HB concentration from 0.69 g/liter to 3.43 g/liter while reducing doubling time from 10.6 h to 8.6 h. The ability to culture and analyze thousands of replicates with high mass transfer in completely mixed culture promises to streamline medium optimization and allow the detection and isolation of highly productive strains. Applications for this system are numerous, encompassing analysis of biofuels and other lipid inclusions, as well as analysis of heterotrophic and photosynthetic systems. PMID:25956771

  2. Addition of antibacterial agents to MMA-TBB dentin bonding systems--influence on tensile bond strength and antibacterial effect.

    PubMed

    Kudou, Y; Obara, K; Kawashima, T; Kubota, M; Abe, S; Endo, T; Komatsu, M; Okuda, R

    2000-03-01

    To produce a bonding system which has both high bond strength and antibacterial properties, an antibacterial agent (vancomycin: VCM or metronidazol: MN) was added to the PMMA powder of 4-META/MMA-TBB resin (CB). The influence of the addition of an antibacterial agent on tensile bond strength to dentin and the antibacterial effect were investigated in this study. Forty-seven freshly extracted bovine first or second incisors were used to measure the tensile bond strength to dentin. The bond strengths to bovine dentin were not significantly decreased by addition of VCM (1%, 2%, 5%), or MN (1%) to CB (p < 0.05). The antibacterial effect of CB containing antibacterial agent on six strains of bacteria was investigated by the agar plate diffusion method, analyzing the appearance of the inhibition zone around a resin disk following anaerobic culturing. The resin disks containing VCM showed antibacterial effects on all of the strains examined; the widths of the inhibition zones were 4-15 mm. The resin disks containing MN showed antibacterial effects on three strains; the widths of the inhibition zones were 0-4 mm. It was thus possible to produce a bonding system with both antibacterial effect and high tensile bond strength by addition of VCM to PMMA powder. PMID:11219091

  3. Design of a Miniature Tissue Culture System to Culture Mouse Heart Valves

    PubMed Central

    Lieber, Samuel C.; Kruithof, Boudewijn P. T.; Aubry, Nadine; Vatner, Stephen F.; Gaussin, Vinciane

    2010-01-01

    Valvular heart disease is a leading cause of morbidity and mortality in adults but little is known about the underlying etiology. A better understanding of the genetic and hemodynamic mechanisms involved in growth and remodeling of heart valves during physiological and pathological conditions is needed for a better understanding of valvular heart disease. Here, we report the design of a miniature tissue culture system (MTCS) that allows the culture of mitral valves from perinatal to adult mice. The design of the MTCS is novel in that fine positioning and cannulation can be conducted with hearts of different sizes (perinatal to adult). Perfusion of the heart and hence, culture of the mitral valve in its natural position, occurs in a hydraulically sealed culture bath environment. Using the MTCS, we successfully cultured the mitral valve of adult mouse hearts for 3 days. Histological analysis indicated that the cultured valves remained viable and their extracellular matrix organization was similar to age-matched native valves. Gene expression could also be modified in cultured valves by perfusion with medium containing beta-galactosidase-expressing adenovirus. Thus, the MTCS is a new tool to study the genetic and hemodynamic mechanisms underlying the three-dimensional organization of the heart valves, which could provide insights in the pathology of valvular heart disease and be used in animal models for the development of tissue-engineered heart valves. PMID:20099034

  4. Plasticity of the Arabidopsis Root System under Nutrient Deficiencies1[C][W][OPEN

    PubMed Central

    Gruber, Benjamin D.; Giehl, Ricardo F.H.; Friedel, Swetlana; von Wirén, Nicolaus

    2013-01-01

    Plant roots show a particularly high variation in their morphological response to different nutrient deficiencies. Although such changes often determine the nutrient efficiency or stress tolerance of plants, it is surprising that a comprehensive and comparative analysis of root morphological responses to different nutrient deficiencies has not yet been conducted. Since one reason for this is an inherent difficulty in obtaining nutrient-deficient conditions in agar culture, we first identified conditions appropriate for producing nutrient-deficient plants on agar plates. Based on a careful selection of agar specifically for each nutrient being considered, we grew Arabidopsis (Arabidopsis thaliana) plants at four levels of deficiency for 12 nutrients and quantified seven root traits. In combination with measurements of biomass and elemental concentrations, we observed that the nutritional status and type of nutrient determined the extent and type of changes in root system architecture (RSA). The independent regulation of individual root traits further pointed to a differential sensitivity of root tissues to nutrient limitations. To capture the variation in RSA under different nutrient supplies, we used principal component analysis and developed a root plasticity chart representing the overall modulations in RSA under a given treatment. This systematic comparison of RSA responses to nutrient deficiencies provides a comprehensive view of the overall changes in root plasticity induced by the deficiency of single nutrients and provides a solid basis for the identification of nutrient-sensitive steps in the root developmental program. PMID:23852440

  5. Microfluidics co-culture systems for studying tooth innervation

    PubMed Central

    Pagella, Pierfrancesco; Neto, Estrela; Jiménez-Rojo, Lucia; Lamghari, Meriem; Mitsiadis, Thimios A.

    2014-01-01

    Innervation plays a key role in the development and homeostasis of organs and tissues of the orofacial complex. Among these structures, teeth are peculiar organs as they are not innervated until later stages of development. Furthermore, the implication of neurons in tooth initiation, morphogenesis and differentiation is still controversial. Co-cultures constitute a valuable method to investigate and manipulate the interactions of nerve fibers with their target organs in a controlled and isolated environment. Conventional co-cultures between neurons and their target tissues have already been performed, but these cultures do not offer optimal conditions that are closely mimicking the in vivo situation. Indeed, specific cell populations require different culture media in order to preserve their physiological properties. In this study we evaluate the usefulness of a microfluidics system for co-culturing mouse trigeminal ganglia and developing teeth. This device allows the application of specific media for the appropriate development of both neuronal and dental tissues. The results show that mouse trigeminal ganglia and teeth survive for long culture periods in this microfluidics system, and that teeth maintain the attractive or repulsive effect on trigeminal neurites that has been observed in vivo. Neurites are repealed when co-cultured with embryonic tooth germs, while postnatal teeth exert an attractive effect to trigeminal ganglia-derived neurons. In conclusion, microfluidics system devices provide a valuable tool for studying the behavior of neurons during the development of orofacial tissues and organs, faithfully imitating the in vivo situation. PMID:25202282

  6. Seven essential strategies for promoting and sustaining systemic cultural competence.

    PubMed

    Delphin-Rittmon, Miriam E; Andres-Hyman, Raquel; Flanagan, Elizabeth H; Davidson, Larry

    2013-03-01

    Racial and ethnic disparities are disturbing facets of the American healthcare system that document the reality of unequal treatment. Research consistently shows that patients of color experience poorer quality of care and health outcomes contributing to increased risks and accelerated mortality rates relative to their white counterparts. While initially conceptualized as an approach for increasing the responsiveness of children's behavioral health care, cultural competence has been adopted as a key strategy for eliminating racial and ethnic health disparities across the healthcare system. However, cultural competence research and practices largely focus on improving provider competencies, while agency and system level approaches for meeting the service needs of diverse populations are given less attention. In this article we offer seven essential strategies for promoting and sustaining organizational and systemic cultural competence. These strategies are to: (1) Provide executive level support and accountability, (2) Foster patient, community and stakeholder participation and partnerships, (3) Conduct organizational cultural competence assessments, (4) Develop incremental and realistic cultural competence action plans, (5) Ensure linguistic competence, (6) Diversify, develop, and retain a culturally competent workforce, and (7) Develop an agency or system strategy for managing staff and patient grievances. For each strategy we offer several recommendations for implementation. PMID:22581030

  7. Multiwell cell culture plate format with integrated microfluidic perfusion system

    NASA Astrophysics Data System (ADS)

    Domansky, Karel; Inman, Walker; Serdy, Jim; Griffith, Linda G.

    2006-01-01

    A new cell culture analog has been developed. It is based on the standard multiwell cell culture plate format but it provides perfused three-dimensional cell culture capability. The new capability is achieved by integrating microfluidic valves and pumps into the plate. The system provides a means to conduct high throughput assays for target validation and predictive toxicology in the drug discovery and development process. It can be also used for evaluation of long-term exposure to drugs or environmental agents or as a model to study viral hepatitis, cancer metastasis, and other diseases and pathological conditions.

  8. Strengths and limitations of the neurosphere culture system.

    PubMed

    Jensen, Josephine B; Parmar, Malin

    2006-12-01

    After the initial reports of free-floating cultures of neural stem cells termed neurospheres (1,2), a wide array of studies using this promising culture system emerged. In theory, this was a near-perfect system for large-scale production of neural cells for use in cell replacement therapies and to assay for and characterize neural stem cells. More than a decade later, after rigorous scrutiny and ample experimental testing of the neurosphere culture system, it has become apparent that the culture system suffers from several disadvantages, and its usefulness is limited for several applications. Nevertheless, the bulk of high-quality research produced over the last decade has also shown that under the right circumstances and for the appropriate purposes, neurospheres hold up to their initial promise. This article discusses the pros and cons of the neurosphere culture system regarding its three major applications: as an assay for neural stem cells, as a model system for neurogenesis and neural development, and for expansion of neural stem cells for transplantation purposes. PMID:17308349

  9. Therapeutically important proteins from in vitro plant tissue culture systems.

    PubMed

    Doran, Pauline M

    2013-01-01

    Plant cells cultured in liquid medium in bioreactors are now being used commercially to produce biopharmaceutical proteins. The emergence of in vitro plant cell culture as a production vehicle reflects the importance of key biosafety and biocontainment concerns affecting the competitiveness of alternative systems such as mammalian cell culture and agriculture. Food plant species are particularly attractive as hosts for in vitro protein production: the risk of transgene escape and food chain contamination is eliminated using containment facilities, while regulatory approval for oral delivery of drugs may be easier than if non-edible species were used. As in whole plants, proteolysis in cultured plant cells can lead to significant degradation of foreign proteins after synthesis; however, substantial progress has been made to counter the destructive effects of proteases in plant systems. Although protein secretion into the culture medium is advantageous for product recovery and purification, measures are often required to minimise extracellular protease activity and product losses due to irreversible surface adsorption. Disposable plastic bioreactors, which are being used increasingly in mammalian cell bioprocessing, are also being adopted for plant cell culture to allow rapid scale-up and generation of saleable product. This review examines a range of technical and regulatory issues affecting the choice of industrial production platform for foreign proteins, and assesses progress in the development of in vitro plant systems for biopharmaceutical production. PMID:23210789

  10. CULTURAL SYSTEM AFFECTS FRUIT QUALITY AND ANTIOXIDANT CAPACITY IN STRAWBERRIES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cultural system [hill plasticulture (HC) vs. matted row (MR)] and genotypes interactions affected strawberry fruit quality. In general, fruit soluble content, total sugar, fructose, glucose, ascorbic acid, titratable acid and citric acid content were increased in the HC system. Fruit from HC also ...

  11. Method and system of culturing an algal mat

    SciTech Connect

    Das, Keshav C; Cannon, Benjamin R; Bhatnagar, Ashish; Chinnasamy, Senthil

    2014-05-13

    A system and method for culturing algae are presented. The system and method utilize a fog of growth medium that is delivered to an algal mat generator along with a stream of CO.sub.2 to promote growth of algal cells contained in the generator.

  12. Cell culture systems to study glial transformation

    SciTech Connect

    Bressler, J.P.; Cole, R.; de Vellis, J.

    1980-01-01

    The transformation of two different types of glial cells has been studied using an in vivo-/in vitro model and a complete in vitro model. The purpose of the study and to define in vitro model systems is to study the the neoplastic transformation of pure populations of glial cells. Data are presented to demonstrate that the transformed cells are glial and tumorigenic. (ACR)

  13. A Hybrid Robotic Control System Using Neuroblastoma Cultures

    NASA Astrophysics Data System (ADS)

    Ferrández, J. M.; Lorente, V.; Cuadra, J. M.; Delapaz, F.; Álvarez-Sánchez, José Ramón; Fernández, E.

    The main objective of this work is to analyze the computing capabilities of human neuroblastoma cultured cells and to define connection schemes for controlling a robot behavior. Multielectrode Array (MEA) setups have been designed for direct culturing neural cells over silicon or glass substrates, providing the capability to stimulate and record simultaneously populations of neural cells. This paper describes the process of growing human neuroblastoma cells over MEA substrates and tries to modulate the natural physiologic responses of these cells by tetanic stimulation of the culture. We show that the large neuroblastoma networks developed in cultured MEAs are capable of learning: establishing numerous and dynamic connections, with modifiability induced by external stimuli and we propose an hybrid system for controlling a robot to avoid obstacles.

  14. Culture and systems of oppression in abused women's lives.

    PubMed

    Phillips, D S

    1998-01-01

    Women's experiences of abuse are shaped by the social and cultural contexts in which they live. Recognition of the complex ways in which culture and systems of oppression interact, creating qualitatively different abuse experiences, is important nursing knowledge. Of particular concern are the ways in which women are constrained in their efforts to combat abuse as they experience the harsh and alienating effects of racism, sexism, classism, and other forms of social injustice. An understanding of the ways in which race and ethnicity, class, language and citizenship, religion, and culture intersect and shape women's experiences of abuse is critical to the provision of culturally competent nursing care. This understanding is the springboard from which more effective assessment and intervention strategies with vulnerable abused women of diverse backgrounds can emerge. PMID:9836163

  15. Horizontally rotated cell culture system with a coaxial tubular oxygenator

    NASA Technical Reports Server (NTRS)

    Wolf, David A. (Inventor); Schwarz, Ray P. (Inventor); Trinh, Tinh T. (Inventor)

    1991-01-01

    The present invention relates to a horizontally rotating bioreactor useful for carrying out cell and tissue culture. For processing of mammalian cells, the system is sterilized and fresh fluid medium, microcarrier beads, and cells are admitted to completely fill the cell culture vessel. An oxygen containing gas is admitted to the interior of the permeable membrane which prevents air bubbles from being introduced into the medium. The cylinder is rotated at a low speed within an incubator so that the circular motion of the fluid medium uniformly suspends the microbeads throughout the cylinder during the cell growth period. The unique design of this cell and tissue culture device was initially driven by two requirements imposed by its intended use for feasibility studies for three dimensional culture of living cells and tissues in space by JSC. They were compatible with microgravity and simulation of microgravity in one G. The vessels are designed to approximate the extremely quiescent low shear environment obtainable in space.

  16. Quantitation of Pseudomonas aeruginosa in wound biopsy samples: from bacterial culture to rapid `real-time' polymerase chain reaction

    PubMed Central

    Pirnay, Jean-Paul; De Vos, Daniel; Duinslaeger, Luc; Reper, Pascal; Vandenvelde, Christian; Cornelis, Pierre; Vanderkelen, Alain

    2000-01-01

    -fold dilutions of the suspension were spread on Luria-Bertani and cetrimide agar plates. Colony counts were performed after overnight incubation at 37°C. The PCR mixture contained sterile distilled water, PCR buffer, deoxynucleotide mixture or digoxigenin labelling mix, MgCl2, diluted template, primers PAL1 and PAL2, and AmpliTaQ DNA polymerase. The amplification was performed in a GeneAmp® PCR System 2400. An aliquot of the reaction mixture was put on an agarose gel for electrophoresis and visualisation of the PCR product. An image of the gel was made using a digital camera. Image analysis software was used to calculate the band mass of the experimental bands. An aliquot of the digoxigenin labelling reaction was denatured and then hybridized with the biotinylated capture probe PrL. Some of the resultant solution was transferred to a well of a streptavidin-coated microtitre plate (MTP) and incubated for 3 h at 45°C. The solution was discarded. Peroxidase conjugated antidigoxigenin was added and the MTP was incubated for 30 min at 37°C. The solution was discarded and ABTS substrate was added. The MTP was incubated for 30 min at 37°C. Absorbance was read at 405 nm. The RTD-PCR mixture contained PCR grade sterile water, diluted template DNA, primers PAL1 and PAL2, 3' fluorescein (FL)-labelled probe oprL-FL, 5' LC Red 640-labelled and 3' phosphorylated probe oprL-LC, MgCl2, and LC DNA Master Hybridisation Probes, containing Taq DNA polymerase, reaction buffer, dNTP mix with dUTP instead of dTTP, and MgCl2. The amplification was performed in a LightCycler™. The fluorescence signal of LC Red 640 was measured during the annealing phase. The measured fluorescence data was processed with analysis software. Results and discussion: The three methods showed a good concordance with the culture results. Conventional PCR was at least 100 times less sensitive than bacterial culture and had a low dynamic range (2 logs). With a lower detection limit of 103 CFU/g tissue, ELISA

  17. The effect of three culture methods on intensive culture system of pacific white shrimp ( Litopenaeus vannamei)

    NASA Astrophysics Data System (ADS)

    Ma, Zhen; Wan, Rong; Song, Xiefa; Gao, Lei

    2013-09-01

    Different culture methods may affect the intensive culture system of Pacific white shrimp ( Litopenaeus vannamei) regarding water quality and growth and economic performance. This study evaluated the potential effects of three culture methods through cultivation of juvenile shrimps under consistent tank management conditions for 84 d. The three methods involved shrimp cultivation in different tanks, i.e., outdoor tanks with cement bottom (mode-C), greenhouse tanks with cement bottom (mode-G) and outdoor tanks with mud-substrate (mode-M). Results showed that water temperature was significantly higher in mode-G than that in mode-C ( P < 0.05). In contrast to the other two treatments, mode-M had stable pH after 50 d cultivation of shrimps. In the mid-late period, the average concentrations of TAN, NO2-N, DIP and COD were significantly lower in mode-M and mode-G compared with those in mode-C ( P < 0.05). Despite lack of differences in the final shrimp weight among different treatments ( P > 0.05), mode-M had significantly higher shrimp yield, survival rate and feed conversion rate ( P < 0.05) than other modes. There were significant differences in revenue and net return among different treatments ( P < 0.05). These demonstrated that the treatments of mode-G and mode-M were conductive to the intensive culture system of L. vannamei.

  18. Optimization of Buffalo (Bubalus bubalis) Embryonic Stem Cell Culture System

    PubMed Central

    Zandi, Mohammad; Muzaffar, Musharifa; Shah, Syed Mohmad; Kumar Singh, Manoj; Palta, Prabhat; Kumar Singla, Suresh; Manik, Radheysham; Chauhan, Manmohan Singh

    2015-01-01

    Objective In order to retain an undifferentiated pluripotent state, embryonic stem (ES) cells have to be cultured on feeder cell layers. However, use of feeder layers limits stem cell research, since experimental data may result from a combined ES cell and feeder cell response to various stimuli. Materials and Methods In this experimental study, a buffalo ES cell line was established from in vitro derived blastocysts and characterized by the Alkaline phosphatase (AP) and immunoflourescence staining of various pluripotency markers. We examined the effect of various factors like fibroblast growth factor 2 (FGF-2), leukemia inhibitory factor (LIF) and Y-27632 to support the growth and maintenance of bubaline ES cells on gelatin coated dishes, in order to establish feeder free culture systems. We also analyzed the effect of feeder-conditioned media on stem cell growth in gelatin based cultures both in the presence as well as in the absence of the growth factors. Results The results showed that Y-27632, in the presence of FGF-2 and LIF, resulted in higher colony growth and increased expression of Nanog gene. Feeder-Conditioned Medium resulted in a significant increase in growth of buffalo ES cells on gelatin coated plates, however, feeder layer based cultures produced better results than gelatin based cultures. Feeder layers from buffalo fetal fibroblast cells can support buffalo ES cells for more than two years. Conclusion We developed a feeder free culture system that can maintain buffalo ES cells in the short term, as well as feeder layer based culture that can support the long term maintenance of buffalo ES cells. PMID:26199905

  19. Moving Towards Culturally Competent Health Systems: Organizational and Market Factors

    PubMed Central

    Weech-Maldonado, Robert; Elliott, Marc; Pradhan, Rohit; Schiller, Cameron; Dreachslin, Janice; Hays, Ron D.

    2012-01-01

    Cultural competency has been proposed as an organizational strategy to address racial/ethnic disparities in the health care system; disparities are a long-standing policy challenge whose relevance is only increasing with the increasing population diversity of the US and across the world. Using an integrative conceptual framework based on the resource dependency and institutional theories, we examine the relationship between organizational and market factors and hospitals’ degree of cultural competency. Our sample consists of 119 hospitals located in the state of California (US) and is constructed using the following datasets for the year 2006: Cultural Competency Assessment Tool of Hospitals (CCATH) Survey, California’s Office of Statewide Health Planning & Development’s Hospital Inpatient Discharges and Annual Hospital Financial Data, American Hospital Association’s Annual Survey, and the Area Resource File. The dependent variable consists of the degree of hospital cultural competency, as assessed by the CCATH overall score. Organizational variables include ownership status, teaching hospital, payer mix, size, system membership, financial performance, and the proportion of inpatient racial/ethnic minorities. Market characteristics included hospital competition, the proportion of racial/ethnic minorities in the area, metropolitan area, and per capita income. Regression analyses were conducted to assess the relationship between the CCATH overall score and organizational and market variables. Our results show that hospitals which are not-for-profit, serve a more diverse inpatient population, and are located in more competitive and affluent markets exhibit a higher degree of cultural competency. Our results underscore the importance of both institutional and competitive market pressures in guiding hospital behavior. For instance, while not-for-profit may adopt innovative/progressive policies like cultural competency simply as a function of their organizational

  20. Voice knowledge acquisition system for the management of cultural heritage

    NASA Astrophysics Data System (ADS)

    Du Château, Stefan; Boulanger, Danielle; Mercier-Laurent, Eunika

    This document presents our work on a definition and experimentation of a voice interface for cultural heritage inventory. This hybrid system includes signal processing, natural language techniques and knowledge modeling for future retrieval. We discuss the first results and give some points on future work.

  1. Cultural Influences on Number Preferences: Christmas and Grading Systems

    ERIC Educational Resources Information Center

    Stieger, Stefan; Krizan, Zlatan

    2013-01-01

    People consistently prefer numbers associated with themselves (e.g., birth dates) over other numbers. We argue that such number preferences are also shaped by cultural influences, such as customs regarding the day on which Christmas is celebrated and customs regarding ranking of numerals used in national school's grading system. Across 6 different…

  2. Parents' Cultural Belief Systems: Their Origins, Expressions, and Consequences.

    ERIC Educational Resources Information Center

    Harkness, Sara, Ed.; Super, Charles M., Ed.

    This volume presents observations and thinking of scholars from a variety of disciplines about parental cultural belief systems. The chapters are concerned with the sources and consequences of parental ethnotheories in a number of societies. The following chapters are included: (1) "Introduction" (Sara Harkness and Charles M. Super); (2) "Parents'…

  3. Barriers to Excellence: The Culture of Silence in School Systems.

    ERIC Educational Resources Information Center

    Rusch, Edith A.

    Little is known about how restructuring networks actually affect the cultures of school systems. This report examines the creation of an "island" of reform in a school district in northwest Ohio. The research emanated from the discovery of a new "island" in northwest Ohio called the Pathfinder Network. The group formed through like-minded…

  4. World Culture in the Capitalist World-System in Transition

    ERIC Educational Resources Information Center

    Griffiths, Tom G.; Arnove, Robert F.

    2015-01-01

    World culture theory (WCT) offers an explanatory framework for macro-level comparative analyses of systems of mass education, including their structures, accompanying policies and their curricular and pedagogical practices. WCT has contributed to broader efforts to overcome methodological nationalism in comparative research. In this paper, we…

  5. Rotating cell culture systems for human cell culture: human trophoblast cells as a model.

    PubMed

    Zwezdaryk, Kevin J; Warner, Jessica A; Machado, Heather L; Morris, Cindy A; Höner zu Bentrup, Kerstin

    2012-01-01

    The field of human trophoblast research aids in understanding the complex environment established during placentation. Due to the nature of these studies, human in vivo experimentation is impossible. A combination of primary cultures, explant cultures and trophoblast cell lines support our understanding of invasion of the uterine wall and remodeling of uterine spiral arteries by extravillous trophoblast cells (EVTs), which is required for successful establishment of pregnancy. Despite the wealth of knowledge gleaned from such models, it is accepted that in vitro cell culture models using EVT-like cell lines display altered cellular properties when compared to their in vivo counterparts. Cells cultured in the rotating cell culture system (RCCS) display morphological, phenotypic, and functional properties of EVT-like cell lines that more closely mimic differentiating in utero EVTs, with increased expression of genes mediating invasion (e.g. matrix metalloproteinases (MMPs)) and trophoblast differentiation. The Saint Georges Hospital Placental cell Line-4 (SGHPL-4) (kindly donated by Dr. Guy Whitley and Dr. Judith Cartwright) is an EVT-like cell line that was used for testing in the RCCS. The design of the RCCS culture vessel is based on the principle that organs and tissues function in a three-dimensional (3-D) environment. Due to the dynamic culture conditions in the vessel, including conditions of physiologically relevant shear, cells grown in three dimensions form aggregates based on natural cellular affinities and differentiate into organotypic tissue-like assemblies. The maintenance of a fluid orbit provides a low-shear, low-turbulence environment similar to conditions found in vivo. Sedimentation of the cultured cells is countered by adjusting the rotation speed of the RCCS to ensure a constant free-fall of cells. Gas exchange occurs through a permeable hydrophobic membrane located on the back of the bioreactor. Like their parental tissue in vivo, RCCS

  6. An Optically Controlled 3D Cell Culturing System

    PubMed Central

    Ishii, Kelly S.; Hu, Wenqi; Namekar, Swapnil A.; Ohta, Aaron T.

    2012-01-01

    A novel 3D cell culture system was developed and tested. The cell culture device consists of a microfluidic chamber on an optically absorbing substrate. Cells are suspended in a thermoresponsive hydrogel solution, and optical patterns are utilized to heat the solution, producing localized hydrogel formation around cells of interest. The hydrogel traps only the desired cells in place while also serving as a biocompatible scaffold for supporting the cultivation of cells in 3D. This is demonstrated with the trapping of MDCK II and HeLa cells. The light intensity from the optically induced hydrogel formation does not significantly affect cell viability. PMID:22701475

  7. INTEGRATED SAFETY MANAGEMENT SYSTEM SAFETY CULTURE IMPROVEMENT INITIATIVE

    SciTech Connect

    MCDONALD JA JR

    2009-01-16

    In 2007, the Department of Energy (DOE) identified safety culture as one of their top Integrated Safety Management System (ISMS) related priorities. A team was formed to address this issue. The team identified a consensus set of safety culture principles, along with implementation practices that could be used by DOE, NNSA, and their contractors. Documented improvement tools were identified and communicated to contractors participating in a year long pilot project. After a year, lessons learned will be collected and a path forward determined. The goal of this effort was to achieve improved safety and mission performance through ISMS continuous improvement. The focus of ISMS improvement was safety culture improvement building on operating experience from similar industries such as the domestic and international commercial nuclear and chemical industry.

  8. Culture and the Immune System: Cultural Consonance in Social Support and C-reactive Protein in Urban Brazil.

    PubMed

    Dressler, William W; Balieiro, Mauro C; Ribeiro, Rosane P; Dos Santos, José Ernesto

    2016-06-01

    In this article, we examine the distribution of a marker of immune system stimulation-C-reactive protein-in urban Brazil. Social relationships are associated with immunostimulation, and we argue that cultural dimensions of social support, assessed by cultural consonance, are important in this process. Cultural consonance is the degree to which individuals, in their own beliefs and behaviors, approximate shared cultural models. A measure of cultural consonance in social support, based on a cultural consensus analysis regarding sources and patterns of social support in Brazil, was developed. In a survey of 258 persons, the association of cultural consonance in social support and C-reactive protein was examined, controlling for age, sex, body mass index, low-density lipoprotein cholesterol, depressive symptoms, and a social network index. Lower cultural consonance in social support was associated with higher C-reactive protein. Implications of these results for future research are discussed. PMID:25828739

  9. Gender and cultural issues in psychiatric nosological classification systems.

    PubMed

    van de Water, Tanya; Suliman, Sharain; Seedat, Soraya

    2016-08-01

    Much has changed since the two dominant mental health nosological systems, the International Classification of Diseases (ICD) and the Diagnostic and Statistical Manual of Mental Disorders (DSM), were first published in 1900 and 1952, respectively. Despite numerous modifications to stay up to date with scientific and cultural changes (eg, exclusion of homosexuality as a disorder) and to improve the cultural sensitivity of psychiatric diagnoses, the ICD and DSM have only recently renewed attempts at harmonization. Previous nosological iterations demonstrate the oscillation in the importance placed on the biological focus, highlighting the tension between a gender- and culture-free nosology (solely biological) and a contextually relevant understanding of mental illness. In light of the release of the DSM 5, future nosological systems, such as the ICD 11, scheduled for release in 2017, and the Research Development Criteria (RDoC), can learn from history and apply critiques. This article aims to critically consider gender and culture in previous editions of the ICD and DSM to inform forthcoming classifications. PMID:27133577

  10. Controlled clinical comparison of three commercial blood culture systems.

    PubMed

    Frank, U; Malkotsis, D; Mlangeni, D; Daschner, F D

    1999-04-01

    In a controlled clinical comparison, three commercial blood culture systems--the standard aerobic BacT/Alert bottle (STD), the aerobic BacT/Alert FAN bottle (FAN) and the Isolator system (ISO; Wampole Laboratories, USA) were compared for their ability to detect aerobic and facultatively anaerobic microorganisms. A total of 945 BacT/Alert (STD and FAN) blood culture sets were compared. Of these, 110 blood culture sets (11.6%) yielded growth of 116 clinically significant bacterial and fungal isolates. Microorganisms were recovered from 10.7% (101/945) of the FAN bottles compared to 8.9% (84/945) of the STD bottles. Of the significant isolates, 78 (67.2%) were recovered by both bottles, 29 (25%) by the FAN bottle only and nine (7.8%) by the STD bottle only (P<0.01). Along with 56.1% (530/945) of BacT/Alert blood culture sets, a concomitant ISO tube was obtained. Of the triple (STD + FAN + ISO) blood culture sets, 54 (10.2%) yielded growth of 59 clinically relevant isolates. Microorganisms were detected in 9.1% (48/530) of the FAN bottles, 8.3% (44/530) of the STD bottles and 4% (21/530) of the ISO tubes (P<0.001). Overall, the BacT/Alert system detected more clinically significant microorganisms than the ISO tube; the STD and the FAN bottle each recovered significantly more staphylococci (P<0.01 and P<0.001, respectively) and gram-negative rods (P<0.01, both). In conclusion, the BacT/Alert FAN bottle performed better than the BacT/Alert STD bottle; both BacT/Alert bottles, however, were superior to the ISO tube in terms of recovery of clinically significant microorganisms, including gram-positive and gram-negative bacteria. PMID:10385012

  11. Information support systems for cultural heritage protection against flooding

    NASA Astrophysics Data System (ADS)

    Nedvedova, K.; Pergl, R.

    2015-08-01

    The goal of this paper is to present use of different kind of software applications to create complex support system for protection of cultural heritage against flooding. The project is very complex and it tries to cover the whole area of the problem from prevention to liquidation of aftermath effects. We used GIS for mapping the risk areas, ontology systems for vulnerability assessment application and the BORM method (Business Object Relation Modelling) for flood protection system planning guide. Those modern technologies helped us to gather a lot of information in one place and provide the knowledge to the broad audience.

  12. Organisational culture matters for system integration in health care.

    PubMed

    Munir, Samina K; Kay, Stephen

    2003-01-01

    This paper illustrates the importance of organisational culture for Clinical Information Systems (CIS) integration. The study is based on data collected in intensive care units in the UK and Denmark. Data were collected using qualitative methods, i.e., observations, interviews and shadowing of health care providers, together with a questionnaire at each site. The data are analysed to extract salient variables for CIS integration, and it is shown that these variables can be separated into two categories that describe the 'Actual Usefulness' of the system and the 'Organisational Culture'. This model is then extended to show that CIS integration directly affects the work processes of the organisation, forming an iterative process of change as a CIS is introduced and integrated. PMID:14728220

  13. An Ex vivo Culture System to Study Thyroid Development

    PubMed Central

    Delmarcelle, Anne-Sophie; Villacorte, Mylah

    2014-01-01

    The thyroid is a bilobated endocrine gland localized at the base of the neck, producing the thyroid hormones T3, T4, and calcitonin. T3 and T4 are produced by differentiated thyrocytes, organized in closed spheres called follicles, while calcitonin is synthesized by C-cells, interspersed in between the follicles and a dense network of blood capillaries. Although adult thyroid architecture and functions have been extensively described and studied, the formation of the “angio-follicular” units, the distribution of C-cells in the parenchyma and the paracrine communications between epithelial and endothelial cells is far from being understood. This method describes the sequential steps of mouse embryonic thyroid anlagen dissection and its culture on semiporous filters or on microscopy plastic slides. Within a period of four days, this culture system faithfully recapitulates in vivo thyroid development. Indeed, (i) bilobation of the organ occurs (for e12.5 explants), (ii) thyrocytes precursors organize into follicles and polarize, (iii) thyrocytes and C-cells differentiate, and (iv) endothelial cells present in the microdissected tissue proliferate, migrate into the thyroid lobes, and closely associate with the epithelial cells, as they do in vivo. Thyroid tissues can be obtained from wild type, knockout or fluorescent transgenic embryos. Moreover, explants culture can be manipulated by addition of inhibitors, blocking antibodies, growth factors, or even cells or conditioned medium. Ex vivo development can be analyzed in real-time, or at any time of the culture by immunostaining and RT-qPCR. In conclusion, thyroid explant culture combined with downstream whole-mount or on sections imaging and gene expression profiling provides a powerful system for manipulating and studying morphogenetic and differentiation events of thyroid organogenesis. PMID:24961920

  14. Culture.

    ERIC Educational Resources Information Center

    1997

    Twelve conference papers on cultural aspects of second language instruction include: "Towards True Multiculturalism: Ideas for Teachers" (Brian McVeigh); Comparing Cultures Through Critical Thinking: Development and Interpretations of Meaningful Observations" (Laurel D. Kamada); "Authority and Individualism in Japan and the USA" (Alisa Woodring);…

  15. A simple method for classification of antibiotics using ion exchange resins added to agar plates.

    PubMed

    Yoshida, K; Kondo, F

    1994-01-01

    Using two different ion-exchange resins (Dowex 50W-X4 as cation and Dowex 1-X4 as anion) added directly to assay plates seeded with Bacillus subtilis or Micrococcus luteus, the size of the inhibitory zone produced by 36 antimicrobial agents around a disc or cup was characterized into various types, such as acidic, basic or amphoteric. An increase of the inhibition zone following addition of 15% Dowex 50W-X4 was evident in penicillins except for ampicillin and penicillin-G, and polyethers. Aminoglycosides, macrolides and colistin, lincomycin, and sulphonamides on assay medium treated with Dowex 1-X4 showed a similar effect on the inhibition zone. Tetracyclines, virginiamycin, oxolinic acid and furazoridone revealed no effects on the inhibition zone with either of the resins. These antibiotics could be divided into various groups on the basis of their chemical structure. This simple and rapid method may be useful for routine laboratory testing of residual antibiotics in meat. PMID:8152391

  16. Development of a selective agar plate for the detection of Campylobacter spp. in fresh produce.

    PubMed

    Yoo, Jin-Hee; Choi, Na-Young; Bae, Young-Min; Lee, Jung-Su; Lee, Sun-Young

    2014-10-17

    This study was conducted to develop a selective medium for the detection of Campylobacter spp. in fresh produce. Campylobacter spp. (n=4), non-Campylobacter (showing positive results on Campylobacter selective agar) strains (n=49) isolated from fresh produce, indicator bacteria (n=13), and spoilage bacteria isolated from fresh produce (n=15) were plated on four Campylobacter selective media. Bolton agar and modified charcoal cefoperazone deoxycholate agar (mCCDA) exhibited higher sensitivity for Campylobacter spp. than did Preston agar and Hunt agar, although certain non-Campylobacter strains isolated from fresh produce by using a selective agar isolation method, were still able to grow on Bolton agar and mCCDA. To inhibit the growth of non-Campylobacter strains, Bolton agar and mCCDA were supplemented with 5 antibiotics (rifampicin, polymyxin B, sodium metabisulfite, sodium pyruvate, ferrous sulfate) and the growth of Campylobacter spp. (n=7) and non-Campylobacter strains (n=44) was evaluated. Although Bolton agar supplemented with rifampicin (BR agar) exhibited a higher selectivity for Campylobacter spp. than did mCCDA supplemented with antibiotics, certain non-Campylobacter strains were still able to grow on BR agar (18.8%). When BR agar with various concentrations of sulfamethoxazole-trimethoprim were tested with Campylobacter spp. (n=8) and non-Campylobacter (n=7), sulfamethoxazole-trimethoprim was inhibitory against 3 of 7 non-Campylobacter strains. Finally, we validated the use of BR agar containing 50mg/L sulfamethoxazole (BRS agar) or 0.5mg/L ciprofloxacin (BRCS agar) and other selective agars for the detection of Campylobacter spp. in chicken and fresh produce. All chicken samples were positive for Campylobacter spp. when tested on mCCDA, BR agar, and BRS agar. In fresh produce samples, BRS agar exhibited the highest selectivity for Campylobacter spp., demonstrating its suitability for the detection of Campylobacter spp. in fresh produce. PMID:25126968

  17. Advanced imaging systems for diagnostic investigations applied to Cultural Heritage

    NASA Astrophysics Data System (ADS)

    Peccenini, E.; Albertin, F.; Bettuzzi, M.; Brancaccio, R.; Casali, F.; Morigi, M. P.; Petrucci, F.

    2014-12-01

    The diagnostic investigations are an important resource in the studies on Cultural Heritage to enhance the knowledge on execution techniques, materials and conservation status of a work of art. In this field, due to the great historical and artistic value of the objects, preservation is the main concern; for this reason, new technological equipment has been designed and developed in the Physics Departments of the Universities of Ferrara and Bologna to enhance the non-invasive approach to the study of pictorial artworks and other objects of cultural interest. Infrared (IR) reflectography, X-ray radiography and computed tomography (CT), applied to works of art, are joined by the same goal: to get hidden information on execution techniques and inner structure pursuing the non-invasiveness of the methods, although using different setup and physical principles. In this work transportable imaging systems to investigate large objects in museums and galleries are presented. In particular, 2D scanning devices for IR reflectography and X-ray radiography, CT systems and some applications to the Cultural Heritage are described.

  18. Gill cell culture systems as models for aquatic environmental monitoring.

    PubMed

    Bury, Nic R; Schnell, Sabine; Hogstrand, Christer

    2014-03-01

    A vast number of chemicals require environmental safety assessments for market authorisation. To ensure acceptable water quality, effluents and natural waters are monitored for their potential harmful effects. Tests for market authorisation and environmental monitoring usually involve the use of large numbers of organisms and, for ethical, cost and logistic reasons, there is a drive to develop alternative methods that can predict toxicity to fish without the need to expose any animals. There is therefore a great interest in the potential to use cultured fish cells in chemical toxicity testing. This review summarises the advances made in the area and focuses in particular on a system of cultured fish gill cells grown into an epithelium that permits direct treatment with water samples. PMID:24574380

  19. An Information System for European culture collections: the way forward.

    PubMed

    Casaregola, Serge; Vasilenko, Alexander; Romano, Paolo; Robert, Vincent; Ozerskaya, Svetlana; Kopf, Anna; Glöckner, Frank O; Smith, David

    2016-01-01

    Culture collections contain indispensable information about the microorganisms preserved in their repositories, such as taxonomical descriptions, origins, physiological and biochemical characteristics, bibliographic references, etc. However, information currently accessible in databases rarely adheres to common standard protocols. The resultant heterogeneity between culture collections, in terms of both content and format, notably hampers microorganism-based research and development (R&D). The optimized exploitation of these resources thus requires standardized, and simplified, access to the associated information. To this end, and in the interest of supporting R&D in the fields of agriculture, health and biotechnology, a pan-European distributed research infrastructure, MIRRI, including over 40 public culture collections and research institutes from 19 European countries, was established. A prime objective of MIRRI is to unite and provide universal access to the fragmented, and untapped, resources, information and expertise available in European public collections of microorganisms; a key component of which is to develop a dynamic Information System. For the first time, both culture collection curators as well as their users have been consulted and their feedback, concerning the needs and requirements for collection databases and data accessibility, utilised. Users primarily noted that databases were not interoperable, thus rendering a global search of multiple databases impossible. Unreliable or out-of-date and, in particular, non-homogenous, taxonomic information was also considered to be a major obstacle to searching microbial data efficiently. Moreover, complex searches are rarely possible in online databases thus limiting the extent of search queries. Curators also consider that overall harmonization-including Standard Operating Procedures, data structure, and software tools-is necessary to facilitate their work and to make high-quality data easily accessible

  20. The Effect of Culture Conditions on Microorganism Identification by Matrix-Assisted Laser Desorption Ionization Mass Spectrometry

    SciTech Connect

    Valentine, Nancy B.; Wunschel, Sharon C.; Wunschel, David S.; Petersen, Catherine E.; Wahl, Karen L.

    2005-01-01

    Abstract Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been used to identify bacteria based upon protein signatures. This research shows that while some different proteins are produced by vegetative bacteria when they are cultured in different growth media, positive identification with MALDI-TOF MS is still possible with the protocol established at Pacific Northwest National Laboratory (PNNL)(11). A core set of small proteins remain constant under at least four different culture media conditions including minimal medium -M9, rich media - tryptic soy broth (TSB) or Luria-Bertani (LB) broth and blood agar plates such that analysis of the intact cells by matrix-assisted laser desorption/ionization mass spectrometry allows for consistent identification.

  1. Use of an insect cell culture growth medium to isolate bacteria from horses with effusive, fibrinous pericarditis: a preliminary study.

    PubMed

    Jones, Samuel L; Valenzisi, Amy; Sontakke, Sushama; Sprayberry, Kimberly A; Maggi, Ricardo; Hegarty, Barbara; Breitschwerdt, Edward

    2007-03-31

    Effusive, fibrinous pericarditis is an uncommon disease entity in horses. In 2001, pericarditis occurred in conjunction with an epizootic in central Kentucky that was associated with exposure to eastern tent caterpillars (ETCs). Bacterial isolation from equine pericardial fluid samples was attempted using an insect cell culture growth medium (ICCGM). Using previously cultured, stored frozen samples from four horses with fibrinous pericarditis, inoculation of 10% blood agar plates yielded no growth, whereas simultaneous inoculation of ICCGM resulted in the isolation of Proprionibacterium acnes, Staphylococcus equorum, a Streptococcus sp. and Pseudomonas rhodesiae from pericardial fluid samples. A similar or novel caterpillar-associated bacteria was not identified; however, use of an ICCGM might enhance isolation of bacteria from equine pericardial fluid. PMID:17204376

  2. TERATOGENICITY OF CYCLOPHOSPHAMIDE IN A COUPLED MICROSOMAL ACTIVATING/EMBRYO CULTURE SYSTEM

    EPA Science Inventory

    Using the coupled microsomal activating/embryo culture system, in vitro experiments were performed to establish the role of metabolism in the embryo toxicity and teratogenicity of cyclophosphamide. Cyclophosphamide in the coupled microsomal activating/embryo culture system produc...

  3. Tissue culture system for infection with human hepatitis delta virus.

    PubMed Central

    Sureau, C; Jacob, J R; Eichberg, J W; Lanford, R E

    1991-01-01

    An in vitro culture system was developed for assaying the infectivity of the human hepatitis delta virus (HDV). Hepatocytes were isolated from chimpanzee liver and grown in a serum-free medium. Cells were shown to be infectible by HDV and to remain susceptible to infection for at least 3 weeks in culture, as evidenced by the appearance of RNA species characteristic of HDV replication as early as 6 days postinfection. When repeated experiments were carried out on cells derived from an animal free of hepatitis B virus (HBV), HDV infection occurred in a consistent fashion but there was no indication of infection with the HBV that was present in the inoculum. Despite numerous attempts with different sources of HBV inocula free of HDV, there was no evidence that indicated susceptibility of these cells to HBV infection. This observation may indicate that HBV and HDV use different modes of entry into hepatocytes. When cells derived from an HBV-infected animal were exposed to HDV, synthesis and release of progeny HDV particles were obtained in addition to HBV replication and production of Dane particles. Although not infectible with HBV, primary cultures of chimpanzee hepatocytes are capable of supporting part of the life cycle of HBV and the entire life cycle of HDV. Images PMID:2041075

  4. The Cultural Erosion Metaphor and the Transcultural Impact of Media Systems.

    ERIC Educational Resources Information Center

    Varan, Duane

    1998-01-01

    Contributes to scholarship on the transcultural impact of media systems (and draws on media effects, political economy, and cultural studies research) by applying the soil erosion metaphor to transcultural impact of television. Discusses four processes associated with this model: cultural abrasion, cultural deflation, cultural deposition, and…

  5. Optimization of culture conditions for an efficient xeno-feeder free limbal cell culture system towards ocular surface regeneration.

    PubMed

    Varghese, Viji Mary; Prasad, Tilak; Kumary, T V

    2010-10-01

    Ex vivo expansion of limbal stem cells from a small biopsy and its subsequent transplantation is the golden choice of treatment for limbal stem cell deficiency. Use of murine 3T3 feeder layer is a prerequisite for this ex vivo expansion. There is an ever-increasing demand for feeder free cultures to avoid xenotoxicity and transmission of xeno-diseases to human system. This study was aimed to establish an efficient xeno-feeder free limbal culture system towards ocular surface regeneration. To study the effect of initial dispase treatment and culture system used, migratory distance of cells from explants was analyzed from phase contrast images using "interactive measurements" of Qwin software (Leica). Expression of p63 in different culture systems was studied by immunofluorescent staining, followed by quantitative confocal microscopy (Carl Zeiss). Results showed dispase treatment was not necessary for establishing limbal explant culture. A combination of Iscove's modified Dulbecco's medium and Panserin 801 resulted in formation of autofeeder layer with maintenance of progenitor characteristics, thus mimicking natural tissue architecture. Further analysis of this culture system showed that cells could be cultured till confluency. Immunofluorescent staining of ABCG2 revealed presence of stem cell marker in the confluent cell layer. Scanning Electron Micrographs demonstrated homogenous population of tightly packed cells in this culture system. Replacement of bovine serum with autologous serum did not affect morphology or growth of cells in this culture system. This study will be a major step in the development of xeno-feeder free epithelial equivalents towards ocular surface reconstruction. PMID:20196106

  6. Modeling human gastrointestinal inflammatory diseases using microphysiological culture systems.

    PubMed

    Hartman, Kira G; Bortner, James D; Falk, Gary W; Ginsberg, Gregory G; Jhala, Nirag; Yu, Jian; Martín, Martín G; Rustgi, Anil K; Lynch, John P

    2014-09-01

    Gastrointestinal illnesses are a significant health burden for the US population, with 40 million office visits each year for gastrointestinal complaints and nearly 250,000 deaths. Acute and chronic inflammations are a common element of many gastrointestinal diseases. Inflammatory processes may be initiated by a chemical injury (acid reflux in the esophagus), an infectious agent (Helicobacter pylori infection in the stomach), autoimmune processes (graft versus host disease after bone marrow transplantation), or idiopathic (as in the case of inflammatory bowel diseases). Inflammation in these settings can contribute to acute complaints (pain, bleeding, obstruction, and diarrhea) as well as chronic sequelae including strictures and cancer. Research into the pathophysiology of these conditions has been limited by the availability of primary human tissues or appropriate animal models that attempt to physiologically model the human disease. With the many recent advances in tissue engineering and primary human cell culture systems, it is conceivable that these approaches can be adapted to develop novel human ex vivo systems that incorporate many human cell types to recapitulate in vivo growth and differentiation in inflammatory microphysiological environments. Such an advance in technology would improve our understanding of human disease progression and enhance our ability to test for disease prevention strategies and novel therapeutics. We will review current models for the inflammatory and immunological aspects of Barrett's esophagus, acute graft versus host disease, and inflammatory bowel disease and explore recent advances in culture methodologies that make these novel microphysiological research systems possible. PMID:24781339

  7. Modeling human gastrointestinal inflammatory diseases using microphysiological culture systems

    PubMed Central

    Hartman, Kira G.; Bortner, James D.; Falk, Gary W.; Ginsberg, Gregory G.; Jhala, Nirag; Yu, Jian; Martín, Martín G.; Rustgi, Anil K.; Lynch, John P.

    2014-01-01

    Gastrointestinal (GI) illnesses are a significant health burden for the US population, with 40 million office visits each year for gastrointestinal complaints and nearly 250,000 deaths. Acute and chronic inflammation are a common element of many GI diseases. Inflammatory processes may be initiated by a chemical injury (acid reflux in the esophagus), an infectious agent (Helicobacter pylori infection in the stomach), autoimmune processes (graft versus host disease after bone marrow transplantation), or idiopathic (as in the case of inflammatory bowel diseases). Inflammation in these settings can contribute to acute complaints (pain, bleeding, obstruction, diarrhea) as well as chronic sequelae including strictures and cancer. Research into the pathophysiology of these conditions has been limited by the availability of primary human tissues or appropriate animal models that attempt to physiologically model the human disease. With the many recent advances in tissue engineering and primary human cell culture systems, it is conceivable that these approaches can be adapted to develop novel human ex vivo systems that incorporate many human cell types to recapitulate in vivo growth and differentiation in inflammatory microphysiological environments. Such an advance in technology would improve our understanding of human disease progression and enhance our ability to test for disease prevention strategies and novel therapeutics. We will review current models for the inflammatory and immunological aspects of Barrett’s esophagus, acute graft versus host disease, and inflammatory bowel disease and explore recent advances in culture methodologies that make these novel microphysiological research systems possible. PMID:24781339

  8. Discussions About “Synthetic Intelligence" in Dissociated Culture System

    NASA Astrophysics Data System (ADS)

    Kudoh, Suguru N.; Kiyohara, Ai; Taguchi, Takahisa

    The fundamental frameworks for possessing qualia are “embodiment” and the network structures of the relationships between internal modules. We proposed “Anaplastic cognitive agent (ACA)” composed by interactions between sub modules with hierarchical history functions and network structures. A dissociated culture system can discriminate several distinct spatiotemporal patterns of action potentials evoked by current inputs, and possesses kinds of history function; a history properties of network dynamics, synaptic plasticity, and so on. These features are fundamental for parts to compose ACA.

  9. Culturally Competent School Leaders: The Individual and the System

    ERIC Educational Resources Information Center

    Hansuvadha, Nat; Slater, Charles L.

    2012-01-01

    Cultural competence is the knowledge, behaviors, and dispositions necessary to effectively interact with other cultural groups. Two case studies are presented which illustrate the cultural competence of administrators in urban settings. Theories are reviewed to investigate the themes of cultural competence that emerged from the professional…

  10. Development of a bovine luteal cell in vitro culture system suitable for co-culture with early embryos.

    PubMed

    Batista, M; Torres, A; Diniz, P; Mateus, L; Lopes-da-Costa, L

    2012-10-01

    The cross talk between the corpus luteum (CL) and the early embryo, potentially relevant to pregnancy establishment, is difficult to evaluate in the in vivo bovine model. In vitro co-culture of bovine luteal cells and early embryos (days 2-8 post in vitro fertilization) may allow the deciphering of this poorly understood cross talk. However, early embryos and somatic cells require different in vitro culture conditions. The objective of this study was to develop a bovine luteal cell in vitro culture system suitable for co-culture with early embryos in order to evaluate their putative steroidogenic and prostanoid interactions. The corpora lutea of the different stages of the estrous cycle (early, mid, and late) were recovered postmortem and enriched luteal cell populations were obtained. In experiments 1 and 2, the effects of CL stage, culture medium (TCM, DMEM-F12, or SOF), serum concentration (5 or 10%), atmosphere oxygen tension (5 or 20%), and refreshment of the medium on the ability of luteal cells to produce progesterone (P(4)) were evaluated. The production of P(4) was significantly increased in early CL cultures, and luteal cells adapted well to simple media (SOF), low serum concentrations (5%), and oxygen tensions (5%). In experiment 3, previous luteal cell cryopreservation did not affect the production of P(4), PGF(2α), and PGE(2) compared to fresh cell cultures. This enables the use of pools of frozen-thawed cells to decrease the variation in cell function associated with primary cell cultures. In experiment 4, mineral oil overlaying culture wells resulted in a 50-fold decrease of the P(4) quantified in the medium, but had no effect on PGF(2α) and PGE(2) quantification. In conclusion, a luteal cell in vitro culture system suitable for the 5-d-long co-culture with early embryos was developed. PMID:23054443

  11. Plant-based culture media: Efficiently support culturing rhizobacteria and correctly mirror their in-situ diversity.

    PubMed

    Youssef, Hanan H; Hamza, Mervat A; Fayez, Mohamed; Mourad, Elhussein F; Saleh, Mohamed Y; Sarhan, Mohamed S; Suker, Ragab M; Eltahlawy, Asmaa A; Nemr, Rahma A; El-Tahan, Mahmod; Ruppel, Silke; Hegazi, Nabil A

    2016-03-01

    Our previous publications and the data presented here provide evidences on the ability of plant-based culture media to optimize the cultivability of rhizobacteria and to support their recovery from plant-soil environments. Compared to the tested chemically-synthetic culture media (e.g. nutrient agar and N-deficient combined-carbon sources media), slurry homogenates, crude saps, juices and powders of cactus (Opuntia ficus-indica) and succulent plants (Aloe vera and Aloe arborescens) were rich enough to support growth of rhizobacteria. Representative isolates of Enterobacter spp., Klebsiella spp., Bacillus spp. and Azospirillum spp. exhibited good growth on agar plates of such plant-based culture media. Cell growth and biomass production in liquid batch cultures were comparable to those reported with the synthetic culture media. In addition, the tested plant-based culture media efficiently recovered populations of rhizobacteria associated to plant roots. Culturable populations of >10(6)-10(8) cfu g(-1) were recovered from the ecto- and endo-rhizospheres of tested host plants. More than 100 endophytic culture-dependent isolates were secured and subjected to morphophysiological identification. Factor and cluster analyses indicated the unique community structure, on species, genera, class and phyla levels, of the culturable population recovered with plant-based culture media, being distinct from that obtained with the chemically-synthetic culture media. Proteobacteria were the dominant (78.8%) on plant-based agar culture medium compared to only 31% on nutrient agar, while Firmicutes prevailed on nutrient agar (69%) compared to the plant-based agar culture media (18.2%). Bacteroidetes, represented by Chryseobacterium indologenes, was only reported (3%) among the culturable rhizobacteria community of the plant-based agar culture medium. PMID:26966571

  12. Plant-based culture media: Efficiently support culturing rhizobacteria and correctly mirror their in-situ diversity

    PubMed Central

    Youssef, Hanan H.; Hamza, Mervat A.; Fayez, Mohamed; Mourad, Elhussein F.; Saleh, Mohamed Y.; Sarhan, Mohamed S.; Suker, Ragab M.; Eltahlawy, Asmaa A.; Nemr, Rahma A.; El-Tahan, Mahmod; Ruppel, Silke; Hegazi, Nabil A.

    2015-01-01

    Our previous publications and the data presented here provide evidences on the ability of plant-based culture media to optimize the cultivability of rhizobacteria and to support their recovery from plant-soil environments. Compared to the tested chemically-synthetic culture media (e.g. nutrient agar and N-deficient combined-carbon sources media), slurry homogenates, crude saps, juices and powders of cactus (Opuntia ficus-indica) and succulent plants (Aloe vera and Aloe arborescens) were rich enough to support growth of rhizobacteria. Representative isolates of Enterobacter spp., Klebsiella spp., Bacillus spp. and Azospirillum spp. exhibited good growth on agar plates of such plant-based culture media. Cell growth and biomass production in liquid batch cultures were comparable to those reported with the synthetic culture media. In addition, the tested plant-based culture media efficiently recovered populations of rhizobacteria associated to plant roots. Culturable populations of >106–108 cfu g−1 were recovered from the ecto- and endo-rhizospheres of tested host plants. More than 100 endophytic culture-dependent isolates were secured and subjected to morphophysiological identification. Factor and cluster analyses indicated the unique community structure, on species, genera, class and phyla levels, of the culturable population recovered with plant-based culture media, being distinct from that obtained with the chemically-synthetic culture media. Proteobacteria were the dominant (78.8%) on plant-based agar culture medium compared to only 31% on nutrient agar, while Firmicutes prevailed on nutrient agar (69%) compared to the plant-based agar culture media (18.2%). Bacteroidetes, represented by Chryseobacterium indologenes, was only reported (3%) among the culturable rhizobacteria community of the plant-based agar culture medium. PMID:26966571

  13. Lean management systems: creating a culture of continuous quality improvement.

    PubMed

    Clark, David M; Silvester, Kate; Knowles, Simon

    2013-08-01

    This is the first in a series of articles describing the application of Lean management systems to Laboratory Medicine. Lean is the term used to describe a principle-based continuous quality improvement (CQI) management system based on the Toyota production system (TPS) that has been evolving for over 70 years. Its origins go back much further and are heavily influenced by the work of W Edwards Deming and the scientific method that forms the basis of most quality management systems. Lean has two fundamental elements--a systematic approach to process improvement by removing waste in order to maximise value for the end-user of the service and a commitment to respect, challenge and develop the people who work within the service to create a culture of continuous improvement. Lean principles have been applied to a growing number of Healthcare systems throughout the world to improve the quality and cost-effectiveness of services for patients and a number of laboratories from all the pathology disciplines have used Lean to shorten turnaround times, improve quality (reduce errors) and improve productivity. Increasingly, models used to plan and implement large scale change in healthcare systems, including the National Health Service (NHS) change model, have evidence-based improvement methodologies (such as Lean CQI) as a core component. Consequently, a working knowledge of improvement methodology will be a core skill for Pathologists involved in leadership and management. PMID:23757036

  14. The Relationship between National Culture and the Usability of an E-Learning System

    ERIC Educational Resources Information Center

    Downey, Steve; Wentling, Rose Mary; Wentling, Tim; Wadsworth, Andrew

    2004-01-01

    This study sought to measure the relationship between national culture and the usability of an e-Learning system by using Hofstede's cultural dimensions and Nielson's usability attributes. The study revealed that high uncertainty avoidance cultures found the system more frustrating to use. The study also revealed that individuals from cultures…

  15. Characterization of nucleoside transport systems in cultured rat epididymal epithelium.

    PubMed

    Leung, G P; Ward, J L; Wong, P Y; Tse, C M

    2001-05-01

    The nucleoside transport systems in cultured epididymal epithelium were characterized and found to be similar between the proximal (caput and corpus) and distal (cauda) regions of the epididymis. Functional studies revealed that 70% of the total nucleoside uptake was Na(+) dependent, while 30% was Na(+) independent. The Na(+)-independent nucleoside transport was mediated by both the equilibrative nitrobenzylthioinosine (NBMPR)-sensitive system (40%) and the NBMPR-insensitive system (60%), which was supported by a biphasic dose response to NBMPR inhibition. The Na(+)-dependent [(3)H]uridine uptake was selectively inhibited 80% by purine nucleosides, indicating that the purine nucleoside-selective N1 system is predominant. Since Na(+)-dependent [(3)H]guanosine uptake was inhibited by thymidine by 20% and Na(+)-dependent [(3)H]thymidine uptake was broadly inhibited by purine and pyrimidine nucleosides, this suggested the presence of the broadly selective N3 system accounting for 20% of Na(+)-dependent nucleoside uptake. Results of RT-PCR confirmed the presence of mRNA for equilibrative nucleoside transporter (ENT) 1, ENT2, and concentrative nucleoside transporter (CNT) 2 and the absence of CNT1. It is suggested that the nucleoside transporters in epididymis may be important for sperm maturation by regulating the extracellular concentration of adenosine in epididymal plasma. PMID:11287319

  16. Biochemical Study of Mixed Culture Prototype in a Closed Ecological System

    NASA Technical Reports Server (NTRS)

    Tischer, R. G.

    1960-01-01

    Since June 1, 1960, the date of initiation of this research project, efforts have been directed toward studying cultural and fermentation patterns and the methodology of pure culture isolation of prototype microorganisms to be employed in closed ecological systems.

  17. A Neuronal Culture System to Detect Prion Synaptotoxicity

    PubMed Central

    Fang, Cheng; Imberdis, Thibaut; Garza, Maria Carmen; Wille, Holger; Harris, David A.

    2016-01-01

    Synaptic pathology is an early feature of prion as well as other neurodegenerative diseases. Although the self-templating process by which prions propagate is well established, the mechanisms by which prions cause synaptotoxicity are poorly understood, due largely to the absence of experimentally tractable cell culture models. Here, we report that exposure of cultured hippocampal neurons to PrPSc, the infectious isoform of the prion protein, results in rapid retraction of dendritic spines. This effect is entirely dependent on expression of the cellular prion protein, PrPC, by target neurons, and on the presence of a nine-amino acid, polybasic region at the N-terminus of the PrPC molecule. Both protease-resistant and protease-sensitive forms of PrPSc cause dendritic loss. This system provides new insights into the mechanisms responsible for prion neurotoxicity, and it provides a platform for characterizing different pathogenic forms of PrPSc and testing potential therapeutic agents. PMID:27227882

  18. Fish Growth in Marine Culture Systems: A Challenge for Biotechnology.

    PubMed

    Lyndon

    1999-07-01

    : Aquaculture production is constrained largely by the growth efficiency of the species being produced. Nutritional approaches have played an important part in improving this situation, but, it is argued, the room for further improvement using such established techniques is limited. Alternative ways of improving fish production by utilizing recent biotechnological advances are explored and assessed as to their potential for commercialization in the near future. Transgenic technologies promise a revolution in aquaculture, but it is considered that consumer resistance may delay the use of transgenic fish for food production. An alternative approach could be the breeding of transgenic fodder plants without the amino acid deficiencies of existing alternatives to fish meal in aquaculture diets. The use of probiotics could reduce antibiotic use on fish farms while they might also provide the basis for "smart" diets, tailored to specific purposes by the inclusion of microorganisms. The selection and genetic engineering of nitrifying and denitrifying bacteria could also pave the way for fully enclosed, recirculating marine culture systems, which would allow control of the environmental variables that currently restrain marine fish culture. PMID:10489415

  19. A Neuronal Culture System to Detect Prion Synaptotoxicity.

    PubMed

    Fang, Cheng; Imberdis, Thibaut; Garza, Maria Carmen; Wille, Holger; Harris, David A

    2016-05-01

    Synaptic pathology is an early feature of prion as well as other neurodegenerative diseases. Although the self-templating process by which prions propagate is well established, the mechanisms by which prions cause synaptotoxicity are poorly understood, due largely to the absence of experimentally tractable cell culture models. Here, we report that exposure of cultured hippocampal neurons to PrPSc, the infectious isoform of the prion protein, results in rapid retraction of dendritic spines. This effect is entirely dependent on expression of the cellular prion protein, PrPC, by target neurons, and on the presence of a nine-amino acid, polybasic region at the N-terminus of the PrPC molecule. Both protease-resistant and protease-sensitive forms of PrPSc cause dendritic loss. This system provides new insights into the mechanisms responsible for prion neurotoxicity, and it provides a platform for characterizing different pathogenic forms of PrPSc and testing potential therapeutic agents. PMID:27227882

  20. Arsenoriboside degradation in marine systems: the use of bacteria culture incubation experiments as model systems.

    PubMed

    Duncan, Elliott G; Maher, William A; Foster, Simon D; Mikac, Katarina M; Krikowa, Frank; Florance, Anthea

    2014-01-01

    Arsenoribosides (as glycerol; phosphate; sulfate and sulfonate) persisted in all bacteria-inoculated cultures irrespective of the source of bacteria (seawater, macro-algae surface) or the culture media used (DIFCO Marine Broth 2216 or novel blended Hormosira banksii tissue-based). This is unlike observations from traditional macro-algae tissue decomposition studies or in nature. In addition known arsenoriboside degradation products such as dimethylarsenoethanol (DMAE), dimethylarsenate (DMA), methylarsenate (MA) and arsenate - As(V) were not detected in any cultures. Consequently, the use of bacterial culture incubation experiments to explain the fate of arsenoribosides in marine systems appears limited as the processes governing arsenoriboside degradation in these experiments appear to be different to those in macro-algae tissue decomposition studies or in nature. PMID:24025537

  1. Mobile system for in-situ imaging of cultural objects

    NASA Astrophysics Data System (ADS)

    Zemlicka, J.; Jakubek, J.; Krejci, F.; Hradil, D.; Hradilova, J.; Mislerova, H.

    2012-01-01

    Non-invasive analytical techniques recently developed with the Timepix pixel detector have shown great potential for the inspection of objects of cultural heritage. We have developed new instrumentation and methodology for in-situ X-ray transmission radiography and X-ray fluorescence imaging and successfully tested and evaluated a mobile system for remote terrain tasks. The prototype portable imaging device comprises the radiation source tube and the spectral sensitive X-ray camera. Both components can be moreover mounted on independent motorized positioning systems allowing adaptation of irradiation geometry to the object shape. Both parts are placed onto a pair of universal portable holders (tripods). The detector is placed in a shielded box with exchangeable entrance window (beam filters and pinhole collimator). This adjustable setup allows performing in-situ measurements for both transmission and emission (XRF) radiography. The assembled system has been successfully tested in our laboratory with phantoms and real samples. The obtained and evaluated results are presented in this paper. Future work will include successive adaptation of the current system for real in-situ utilization and preparation of software allowing semi-automatic remote control of measurements.

  2. Competition of Invertebrates Mixed Culture in the Closed Aquatic System

    NASA Astrophysics Data System (ADS)

    Pisman, Tamara

    The study considers the experimental model of interactions between invertebrates (the cilates Paramecium caudatum, Paramecium bursaria and the rotifers Brachionis plicatilis) in the closed aquatic system. The infusoria P.caudatum can feed on yeast, bacteria and chlorella; in this experiment growth and reproduction were maintained by bacteria only. The P.bursaria - zoochlorella endosymbiosis is a natural model of a simple biotic cycle. P.bursaria consumes glucose and oxygen released by zoochlorella in the process of biosynthesis and releases nitrogenous compounds and carbon dioxide necessary for algal photosynthesis. The rotifers Br. plicatilis can consume algae, bacteria and detritus. Thus in experiment with the mixed culture of invertebrates they can use different food sources. However with any initial percentage of the invertebrates the end portion of P.bursaria reaches 90-99

  3. Organotypic slice co-culture systems to study axon regeneration in the dopaminergic system ex vivo.

    PubMed

    Heine, Claudia; Franke, Heike

    2014-01-01

    Organotypic slice co-cultures are suitable tools to study axonal regeneration and development (growth or regrowth) of different projection systems of the CNS under ex vivo conditions.In this chapter, we describe in detail the reconstruction of the mesocortical and nigrostriatal dopaminergic projection system culturing tissue slices from the ventral tegmental area/substantia nigra (VTA/SN) with the prefrontal cortex (PFC) or the striatum (STR). The protocol includes the detailed slice preparation and incubation. Moreover, different application possibilities of the ex vivo model are mentioned; as an example, the substance treatment procedure and biocytin tracing are described to reveal the effect of applied substances on fiber outgrowth. PMID:24838961

  4. PCR amplification of Bartonella koehlerae from human blood and enrichment blood cultures

    PubMed Central

    2010-01-01

    Background Cats appear to be the primary reservoir host for Bartonella koehlerae, an alpha Proteobacteria that is most likely transmitted among cat populations by fleas (Ctenocephalides felis). Bartonella koehlerae has caused endocarditis in a dog and in one human patient from Israel, but other clinically relevant reports involving this bacterium are lacking. Despite publication of numerous, worldwide epidemiological studies designed to determine the prevalence of Bartonella spp. bacteremia in cats, B. koehlerae has never been isolated using conventional blood agar plates. To date, successful isolation of B. koehlerae from cats and from the one human endocarditis patient has consistently required the use of chocolate agar plates. Results In this study, Bartonella koehlerae bacteremia was documented in eight immunocompetent patients by PCR amplification and DNA sequencing, either prior to or after enrichment blood culture using Bartonella alpha Proteobacteria growth medium. Presenting symptoms most often included fatigue, insomnia, joint pain, headache, memory loss, and muscle pain. Four patients were also infected with Bartonella vinsonii subsp. berkhoffii genotype II. After molecular documentation of B. koehlerae infection in these patients, a serological test was developed and serum samples were tested retrospectively. Bartonella koehlerae antibodies were not detected (titers < 1:16) in 30 healthy human control sera, whereas five of eight patient samples had B. koehlerae antibody titers of 1:64 or greater. Conclusions Although biased by a study population consisting of individuals with extensive arthropod and animal exposure, the results of this study suggest that B. koehlerae bacteremia is more common in immunocompetent people than has been previously suspected. Future studies should more thoroughly define modes of transmission and risk factors for acquiring infection with B. koehlerae. In addition, studies are needed to determine if B. koehlerae is a cause or

  5. Three-dimensional systems for in vitro follicular culture: overview of alginate-based matrices.

    PubMed

    Brito, Ivina R; Lima, Isadora M T; Xu, Min; Shea, Lonnie D; Woodruff, Teresa K; Figueiredo, José R

    2014-08-01

    The in vitro culture of ovarian follicles has provided critical insight into the biology of the follicle and its enclosed oocyte and the physical interaction and communication between the theca and granulosa cells and the oocyte that is necessary to produce meiotically competent oocytes. Various two-dimensional (2D) and three-dimensional (3D) culture systems have been developed to evaluate the effect of growth factors, hormones, extracellular matrix components and culture conditions on follicle development and oocyte growth and maturation. Among these culture systems, 3D systems make it possible to maintain follicle structure and support communication between the various cell compartments within the follicle. In this review article, we will discuss the three main approaches to ovarian follicle culture: 2D attachment systems, 3D floating systems and 3D encapsulated systems. We will specifically emphasise the development of and advances in alginate-based encapsulated systems for in vitro follicle culture. PMID:23866836

  6. Culturing Human Pluripotent and Neural Stem Cells in an Enclosed Cell Culture System for Basic and Preclinical Research

    PubMed Central

    Stover, Alexander E.; Herculian, Siranush; Banuelos, Maria G.; Navarro, Samantha L.; Jenkins, Michael P.; Schwartz, Philip H.

    2016-01-01

    This paper describes how to use a custom manufactured, commercially available enclosed cell culture system for basic and preclinical research. Biosafety cabinets (BSCs) and incubators have long been the standard for culturing and expanding cell lines for basic and preclinical research. However, as the focus of many stem cell laboratories shifts from basic research to clinical translation, additional requirements are needed of the cell culturing system. All processes must be well documented and have exceptional requirements for sterility and reproducibility. In traditional incubators, gas concentrations and temperatures widely fluctuate anytime the cells are removed for feeding, passaging, or other manipulations. Such interruptions contribute to an environment that is not the standard for cGMP and GLP guidelines. These interruptions must be minimized especially when cells are utilized for therapeutic purposes. The motivation to move from the standard BSC and incubator system to a closed system is that such interruptions can be made negligible. Closed systems provide a work space to feed and manipulate cell cultures and maintain them in a controlled environment where temperature and gas concentrations are consistent. This way, pluripotent and multipotent stem cells can be maintained at optimum health from the moment of their derivation all the way to their eventual use in therapy. PMID:27341536

  7. Culturing Human Pluripotent and Neural Stem Cells in an Enclosed Cell Culture System for Basic and Preclinical Research.

    PubMed

    Stover, Alexander E; Herculian, Siranush; Banuelos, Maria G; Navarro, Samantha L; Jenkins, Michael P; Schwartz, Philip H

    2016-01-01

    This paper describes how to use a custom manufactured, commercially available enclosed cell culture system for basic and preclinical research. Biosafety cabinets (BSCs) and incubators have long been the standard for culturing and expanding cell lines for basic and preclinical research. However, as the focus of many stem cell laboratories shifts from basic research to clinical translation, additional requirements are needed of the cell culturing system. All processes must be well documented and have exceptional requirements for sterility and reproducibility. In traditional incubators, gas concentrations and temperatures widely fluctuate anytime the cells are removed for feeding, passaging, or other manipulations. Such interruptions contribute to an environment that is not the standard for cGMP and GLP guidelines. These interruptions must be minimized especially when cells are utilized for therapeutic purposes. The motivation to move from the standard BSC and incubator system to a closed system is that such interruptions can be made negligible. Closed systems provide a work space to feed and manipulate cell cultures and maintain them in a controlled environment where temperature and gas concentrations are consistent. This way, pluripotent and multipotent stem cells can be maintained at optimum health from the moment of their derivation all the way to their eventual use in therapy. PMID:27341536

  8. A three-dimensional culture system using alginate hydrogel prolongs hatched cattle embryo development in vitro.

    PubMed

    Zhao, Shuan; Liu, Zhen-Xing; Gao, Hui; Wu, Yi; Fang, Yuan; Wu, Shuai-Shuai; Li, Ming-Jie; Bai, Jia-Hua; Liu, Yan; Evans, Alexander; Zeng, Shen-Ming

    2015-07-15

    No successful method exists to maintain the three-dimensional architecture of hatched embryos in vitro. Alginate, a linear polysaccharide derived from brown algae, has characteristics that make it an ideal material as a three-dimensional (3D) extracellular matrix for in vitro cell, tissue, or embryo culture. In this study, alginate hydrogel was used for IVC of posthatched bovine embryos to observe their development under the 3D system. In vitro-fertilized and parthenogenetically activated posthatched bovine blastocysts were cultured in an alginate encapsulation culture system (AECS), an alginate overlay culture system (AOCS), or control culture system. After 18 days of culture, the survival rate of embryos cultured in AECS was higher than that in the control group (P < 0.05), and the embryos were expanded and elongated in AECS with the maximal length of 1.125 mm. When the AECS shrinking embryos were taken out of the alginate beads on Day 18 and cultured in the normal culture system, 9.09% of them attached to the bottoms of the plastic wells and grew rapidly, with the largest area of an attached embryo being 66.00 mm(2) on Day 32. The embryos cultured in AOCS developed monovesicular or multivesicular morphologies. Total cell number of the embryos cultured in AECS on Day 19 was significantly higher than that of embryos on Day 8. Additionally, AECS and AOCS supported differentiation of the embryonic cells. Binuclear cells were visible in Day-26 adherent embryos, and the messenger RNA expression patterns of Cdx2 and Oct4 in AOCS-cultured embryos were similar to those in vivo embryos, whereas IFNT and ISG15 messenger RNA were still expressed in Day-26 and Day-32 prolong-cultured embryos. In conclusion, AECS and AOCS did support cell proliferation, elongation, and differentiation of hatched bovine embryos during prolonged IVC. The culture system will be useful to further investigate the molecular mechanisms controlling ruminant embryo elongation and implantation. PMID

  9. Microbial Mediation of Dolomite Precipitation in Natural Environments, Culture Experiments and Molecular Studies

    NASA Astrophysics Data System (ADS)

    Meister, P.; Nealson, K.; McKenzie, J. A.; Warthmann, R.; Vasconcelos, C.

    2005-12-01

    Although dolomite [CaMg(CO3)2] is a common carbonate mineral in sedimentary rocks, it is rarely observed forming in modern environments, and, until recently, experimental precipitation under Earth surface conditions proved impossible. With the discovery of microbial mediated dolomite formation in culture experiments with sulfate-reducing bacteria, it has become apparent that microbes play an important role in overcoming the kinetic barrier of mineral precipitation and, thus, may represent a key factor controlling early diagenetic processes throughout Earth history. The detailed mechanisms of these processes, however, remain poorly understood. Recent studies of dolomite layers in organic carbon-rich hemipelagic sediments recovered on the Peru margin during Ocean Drilling Program Leg 201 (Meister et al., in prep.) indicate precipitation at the interface between the sulphate reduction and methanogenic zones. At this chemical front, alkalinity is strongly increased, sulphate ions, a possible inhibitor of dolomite precipitation, are efficiently removed, and highest total cell densities were counted (up to 10 to the 9 cells / cm3; Shipboard Scientific Party, 2003). These results strengthen the model that microbes are involved in dolomite formation, providing the appropriate chemical conditions, whereas the high cell density may kinetically control the strictly focused precipitation process. We are currently conducting a systematic study of the precipitation of dolomite and other carbonate minerals in the Ca-Mg-bicarbonate-system. In preliminary experiments under aerobic conditions we used agar plates with a marine medium to grow a bacterium isolated from sediments of the San Pedro basin (California), an upwelling area similar to the Peru margin. We observed that the crystals formed only inside of the colonies and showed a dumbbell-shaped morphology similar to dolomite produced in anaerobic experiments. X-ray diffraction patterns revealed, however, that the product was

  10. Multispectral light metering system for cultural heritage diagnosis and conservation

    NASA Astrophysics Data System (ADS)

    Miccoli, Matteo; Melis, Marcello

    2013-05-01

    In the world of Cultural Heritage the first concern is all about Conservation of the works of art. A piece of art in bad shape is meant to deteriorate to an irreversible stage. To avoid this, quite often it's needed to go through one or more cycles of restoration to clean and consolidate the various elements of the piece. The very second concern, once the work of art is restored and in good and stable shape, is its fruition. At the end of the day why one should do all that restoration work if nobody then can access and view? Yet viewing and enjoying an artwork means that a visitor would be able to see it at its best, and this means, almost always, to have a good lighting system. Today, both restoration and fruition can greatly benefit of all the available technologies, and achieve very high level quality. The goal of this paper is the development of an exposimetric system suitable to be extremely useful as a tool for the the non invasive analysis, as well as for the lighting design and lighting systems monitoring. Many diagnosis techniques that are used before the restoration stage, require a suitable lighting system to allow to extract from the painting the maximum amount of information through the acquisition of images in the range of visible as well as UV and IR light. A standard exposimeter is for its own nature, sensitive only to the visible light, constrained by the standard photometric sensitivity curve V(lambda). A wide band exposimeter would be, on the other hand, an invaluable tool to get higher precision and to speed up multispectral wide band images acquisition, avoiding time wasting fail and try cycles to record the subject under wide spectrum conditions. The same equipment can be used to monitor the quality of the light in a expo lighting system at, for example, a museum or a gallery. The light hitting a piece of art has to allow the visitor to see and appreciate all the color shades, and to appreciate the contrast of dark and bright areas due only to

  11. Bacillus polymachus sp. nov., with a broad range of antibacterial activity, isolated from forest topsoil samples by using a modified culture method.

    PubMed

    Nguyen, Tuan Manh; Kim, Jaisoo

    2015-02-01

    A new, modified culture method that utilizes a transwell plate with a 0.4 µm pore-size microporous membrane was developed. This system allows only trace nutrients from the soil into the liquid culture through the microporous membrane. The method is a more powerful tool for the discovery of novel species from soils than are traditional methods. Such newly identified species could potentially produce useful metabolites. A bacterial strain, T515(T), was isolated using this modified culture method. Growth of strain T515(T) occurred at pH 4-9 in a temperature range between 20 °C and 40 °C and in the presence of 0-2 % (w/v) NaCl on R2A agar. Colonies on the agar plates were tiny, white, and convex after 5 days incubation at 28 °C. Comparative analysis of the nearly full-length 16S rRNA gene sequence of strain T515(T) revealed close pairwise similarity with species of the genus Bacillus, and strain T515(T) was most closely related to Bacillus panaciterrae Gsoil 1517(T) (96.7 %) and Bacillus funiculus NAF001(T) (96.0 %). The major quinone of strain T515(T) was menaquinone-7 (MK-7) and the major fatty acids were iso-C15 : 0 (45.5 %), anteiso-C15 : 0 (23.2 %) and C16 : 0 (10.9 %). The predominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Strain T515(T) was sensitive to streptomycin and tetracycline, but resistant to rifampicin (0.125 µg ml(-1)), ampicillin (0.5 µg ml(-1)) and chloramphenicol (1 µg ml(-1)). The strain showed antimicrobial activities against the six strains tested: Bacillus subtilis KEMB 51201-001, Staphylococcus aureus KEMB 4659, Pseudomonas aeruginosa KACC 10185, Staphylococcus epidermidis KACC 13234, Paenibacillus larvae KACC 14031 and Escherichia coli KEMB 212-234. Based on these results, strain T515(T) represents a novel species of the genus Bacillus with the proposed name, Bacillus polymachus sp. nov. The type strain is T515(T) ( = KEMB 9005-168(T) = KACC 18242(T) = NBRC 110614(T)). PMID

  12. Timely culture for mycobacteria which utilizes a microcolony method.

    PubMed Central

    Welch, D F; Guruswamy, A P; Sides, S J; Shaw, C H; Gilchrist, M J

    1993-01-01

    For the isolation of mycobacteria from clinical specimens, we evaluated a method that used a thinly poured Middlebrook 7H11 agar plate (10 by 90 mm) that was examined microscopically. Inoculated plates were sealed, incubated, and examined at regular intervals for the appearance of microcolonies. Plates were examined microscopically, while still sealed, by focusing on the agar surface through the bottom of the plate and the agar. Plates were scanned at low power (x40 total magnification), and colony morphology was confirmed at intermediate power (x100 to x180 magnification). This method was compared with a traditional method that used macroscopic examination of standard mycobacterial media. By using all specimens submitted for mycobacterial culture over the duration of the study, the method was evaluated until 270 isolates of mycobacteria (Mycobacterium tuberculosis, n = 103; M. avium-M. intracellulare, n = 115; miscellaneous, n = 52) were detected. While the conventional method required an average of 23 days to the time of first detection of mycobacteria, the experimental method required an average of only 11 days. When limited to acid-fast stain-positive specimens that were culture positive for M. tuberculosis, the average interval to positivity was 7 days for the microcolony method compared with 17 days for the conventional method. With the experimental method, the microscopic colonial morphology allowed for the presumptive identification of M. tuberculosis colonies, which were distinguished by cording, and M. avium-M. intracellulare colonies, which were smooth and entire. Presumptive identification was complete for 83.5% of the M. tuberculosis isolates within 10 days and for 85% of the M. avium-M. intracellulare isolates within 11 days after inoculation. If the microcolony method was combined with a conventional tube medium, the composite would optimize for speed of recovery while providing the full sensitivity of the conventional method. In addition to reducing

  13. A model for culturally adapting a learning system.

    PubMed

    Del Rosario, M L

    1975-12-01

    The Cross-Cultural Adaption Model (XCAM) is designed to help identify cultural values contained in the text, narration, or visual components of a learning instrument and enables the adapter to evaluate his adapted model so that he can modify or revise it, and allows him to assess the modified version by actually measuring the amount of cultural conflict still present in it. Such a model would permit world-wide adaption of learning materials in population regulation. A random sample of the target group is selected. The adapter develops a measurin g instrument, the cross-cultural adaption scale (XCA), a number of statements about the cultural affinity of the object evaluated. The pretest portion of the sample tests the clarity and understandability of the rating scale to be used for evaluating the instructional materials; the pilot group analyzes the original version of the instructional mater ials, determines the criteria for change, and analyzes the adapted version in terms of these criteria; the control group is administered the original version of the learning materials; and the experimental group is administered the adapted version. Finally, the responses obtained from the XRA rating scale and discussions of both the experimental and control groups are studied and group differences are ev aluated according to cultural conflicts met with each version. With this data, the preferred combination of elements is constructed. PMID:12307758

  14. RAT TRACHEAL CELL CULTURE TRANSFORMATION SYSTEM FOR ASSESSMENT OF ENVIRONMENTAL AGENTS AS CARCINOGENS AND PROMOTERS

    EPA Science Inventory

    A tracheal cell culture system which can be used for detection of hazardous environmental agents is described. The culture system makes use of primary tracheal cells that are isolated from rats by protease digestion of the tracheal epithelium. The epithelial cells are plated on a...

  15. Revisiting the Role of Cultural Capital in East Asian Educational Systems: The Case of South Korea

    ERIC Educational Resources Information Center

    Byun, Soo-yong; Schofer, Evan; Kim, Kyung-keun

    2012-01-01

    The concept of cultural capital has proved invaluable in understanding educational systems in Western countries, and recent work seeks to extend those insights to the diverse educational systems of other geographic regions. Using data from the 2000 Programme for International Student Assessment, the authors explored cultural capital in South Korea…

  16. Adult Educators and Cultural Competence within Health Care Systems: Change at the Individual and Structural Levels

    ERIC Educational Resources Information Center

    Ziegahn, Linda; Ton, Hendry

    2011-01-01

    Goals of cultural competence are commonly described as creation of a health care system and workforce capable of delivering high-quality care to all patients regardless of race, ethnicity, culture, or language. While this "system" is made up of individuals, it also has a life of its own, as with all institutions. In this chapter, the authors…

  17. Potential of fungal co-culturing for accelerated biodegradation of petroleum hydrocarbons in soil.

    PubMed

    Yanto, Dede Heri Yuli; Tachibana, Sanro

    2014-08-15

    The potential of fungal co-culture of the filamentous Pestalotiopsis sp. NG007 with four different basidiomycetes--Trametes versicolor U97, Pleurotus ostreatus PL1, Cerena sp. F0607, and Polyporus sp. S133--for accelerating biodegradation of petroleum hydrocarbons (PHCs) was studied using three different physicochemical characteristic PHCs in soil. All the combinations showed a mutual intermingling mycelial interaction on the agar plates. However, only NG007/S133 (50/50) exhibited an optimum growth rate and enzymatic activities that supported the degradation of asphalt in soil. The co-culture also degraded all fractions at even higher concentrations of the different PHCs. In addition, asphaltene, which is a difficult fraction for a single microorganism to degrade, was markedly degraded by the co-culture, which indicated that the simultaneous biodegradation of aliphatic, aromatic, resin, and asphaltene fractions had occurred in the co-culture. An examination of in-vitro degradation by the crude enzymes and the retrieval fungal culture from the soil after the experiment confirmed the accelerated biodegradation due to enhanced enzyme activities in the co-culture. The addition of piperonyl butoxide or AgNO3 inhibited biodegradation by 81-99%, which demonstrated the important role of P450 monooxygenases and/or dioxygenases in the initial degradation of the aliphatic and aromatic fractions in PHCs. PMID:24997261

  18. Nitric oxide delivery system for cell culture studies.

    PubMed

    Wang, Chen; Deen, William M

    2003-01-01

    To investigate the toxicity and mutagenicity of NO, methods are needed to deliver it to cell cultures at known, constant rates. To permit continuous exposures over lengthy periods, we fabricated a simple apparatus utilizing gas-permeable polydimethylsiloxane (Silastic) tubing to supply both NO and O2 to a stirred, cylindrical vessel. Mass transfer in this system was characterized by measuring the delivery rates of NO or O2 alone, and of NO to air-saturated solutions. The concentrations of NO, O2, and NO2- (the end product of NO oxidation) were monitored continuously. The total flux of nitrogen species into the liquid (as determined from the sum of NO and NO2- accumulation) was 50%-90% greater in the presence of O2, depending on the NO partial pressure in the gas. Also, the simultaneously measured mass transfer coefficients for NO and O2 differed greatly from the corresponding unreactive values. An analysis of the data using diffusion-reaction models showed that NO oxidation in the aqueous boundary layer contributed very little to the nitrogen flux increase or to variations in the mass transfer coefficients. However, the unusually strong dependence of the delivery rates on chemical reactions could be explained by postulating that partial oxidation of NO to NO2 occurred within the membrane. The rate constant we estimated for polydimethylsiloxane, 4.4 x 10(5) M-2 s(-1) at 23 degrees C, is only about one-fifth of values reported previously for water and nonpolar solvents, but the high solubilities of NO and O2 in the polymer are sufficient to make NO2 formation significant. Although considerable NO2 is calculated to enter the liquid, its reaction with aqueous NO is rapid enough to keep this undesired compound at trace levels, except within a few microns of the tubing. Thus, cells will have little exposure to NO2 PMID:12572657

  19. An analysis of the productivity of a CELSS continuous algal culture system

    NASA Technical Reports Server (NTRS)

    Radmer, R.; Behrens, P.; Fernandez, E.; Arnett, K.

    1986-01-01

    One of the most attractive aspects of using algal cultures as plant components for a Closed Ecological Life Support Systems (CELSS) is the efficiency with which they can be grown. Although algae are not necessarily intrinsically more efficient than higher plants, the ease which they can be handled and manipulated (more like chemical reagents than plants), and the culturing techniques available, result in much higher growth rates than are usually attainable with higher plants. Furthermore, preliminary experiments have demonstrated that algal growth and physiology is not detectable altered in a microgravity environment, (1) whereas the response of higher plants to zero gravity is unknown. In order to rationally design and operate culture systems, it is necessary to understand how the macroparameters of a culture system, e.g., productivity, are related to the physiological aspects of the algal culture. A first principles analysis of culture system is discussed, and a mathematical model that describes the relationship of culture productivity to the cell concentration of light-limited culture is derived. The predicted productivity vs cell concentration curve agrees well with the experimental data obtained to test this model, indicating that this model permits an accurate prediction of culture productivity given the growth parameters of the system.

  20. A novel culture system for adult porcine intestinal crypts.

    PubMed

    Khalil, Hassan A; Lei, Nan Ye; Brinkley, Garrett; Scott, Andrew; Wang, Jiafang; Kar, Upendra K; Jabaji, Ziyad B; Lewis, Michael; Martín, Martín G; Dunn, James C Y; Stelzner, Matthias G

    2016-07-01

    Porcine models are useful for investigating therapeutic approaches to short bowel syndrome and potentially to intestinal stem cell (ISC) transplantation. Whereas techniques for the culture and genetic manipulation of ISCs from mice and humans are well established, similar methods for porcine stem cells have not been reported. Jejunal crypts were isolated from murine, human, and juvenile and adult porcine small intestine, suspended in Matrigel, and co-cultured with syngeneic intestinal subepithelial myofibroblasts (ISEMFs) or cultured without feeder cells in various culture media. Media containing epidermal growth factor, noggin, and R-spondin 1 (ENR medium) were supplemented with various combinations of Wnt3a- or ISEMF-conditioned medium (CM) and with glycogen synthase kinase 3 inhibitor (GSK3i), and their effects were studied on cultured crypts. Cell lineage differentiation was assessed by immunohistochemistry and quantitative polymerase chain reaction. Cultured porcine cells were serially passaged and transduced with a lentiviral vector. Whereas ENR medium supported murine enteroid growth, it did not sustain porcine crypts beyond 5 days. Supplementation of Wnt3a-CM and GSK3i resulted in the formation of complex porcine enteroids with budding extensions. These enteroids contained a mixture of stem and differentiated cells and were successfully passaged in the presence of GSK3i. Crypts grown in media supplemented with porcine ISEMF-CM formed spheroids that were less well differentiated than enteroids. Enteroids and spheroids were transfected with a lentivirus with high efficiency. Thus, our method maintains juvenile and adult porcine crypt cells long-term in culture. Porcine enteroids and spheroids can be successfully passaged and transduced by using lentiviral vectors. PMID:26928041

  1. Establishment of banking system for allogeneic cultured dermal substitute.

    PubMed

    Kuroyanagi, Yoshimitsu; Kubo, Kentaro; Matsui, Hiromich; Kim, Hyun Jung; Numari, Shinichiro; Mabuchi, Yho; Kagawa, Shizuko

    2004-01-01

    Allogeneic cultured dermal substitute (CDS) was prepared by culturing fibroblasts on a two-layered spongy matrix of hyaluronic acid (HA) and atelo-collagen (Col). Allogeneic CDS can be cryopreserved and transported to other hospitals in a frozen state. Vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), platelet derived growth factor (PDGF)-AA, transforming growth factor (TGF)-beta1, keratinocytes growth factor (KGF), interleukin (IL)-6 and IL-8 were contained in the culture medium which was used in preparing CDS over a cultivation period of one week (fresh CDS culture medium sample). After thawing a cryopreserved CDS, the CDS was recultured in a culture medium for one week. VEGF, bFGF, HGF, TGF-beta1 and IL-8 were contained in the culture medium which was used in reculturing CDS for one week (cryopreserved CDS culture medium sample), although some cytokines were detected at a lower level than those before freezing. This finding suggests that the cryopreserved CDS retains its ability to release these cytokines. Clinical research on allogeneic CDS, which was newly developed at the R & D Center for Artificial Skin of Kitasato University, has been carried out in medical centers across Japan with the support of the Millennium Project of the Ministry of Health, Labor and Welfare. It was demonstrated that the allogeneic CDS functions as an excellent cell therapy for intractable skin ulcers as well as burn injuries. The spongy matrix itself, as well as the cytokines released from the allogeneic CDS, seemed to be beneficial for the treatment of intractable skin defect. PMID:14720283

  2. Multifunctional encoding system for assessment of movable cultural heritage

    NASA Astrophysics Data System (ADS)

    Tornari, V.; Bernikola, E.; Osten, W.; Groves, R. M.; Marc, G.; Hustinx, G. M.; Kouloumpi, E.; Hackney, S.

    2007-07-01

    This is an introductory paper of a recent EC project dealing with research in cultural heritage and aiming to communicate new fields of application for optical metrology techniques. The project is in its initial state and more conclusive information is expected to be available at the time of the perspective conference. Nowadays safety, ethical, economical and security issues as well as the increase demand for loaning of art objects for exhibitions in transit, are forcing the Conservation Community to undertake strong initiatives and actions against various types of mistreatment, damage or fraud, during transportation of movable Cultural Heritage. Therefore the interest directs to the development of innovative methodologies and instrumentation to respond to critical aspects of increased importance in cultural heritage preservation, among which of prior consideration are: to secure proper treatment, assess probable damage, fight fraud actions in transportation.

  3. Teratological research using in vitro systems. I. Mammalian whole embryo culture.

    PubMed Central

    Flynn, T J

    1987-01-01

    Approximately 390 literature references (through spring 1986) were reviewed for mammalian whole embryo culture procedures, with particular attention to the development of those cultures as systems for teratogenicity testing. The existing procedures could be conveniently divided into three groups, which are defined by the periods of embryogenesis that they embrace: preimplantation, peri-implantation, and post-implantation culture systems. The literature on peri-implantation embryo culture was sparse, and it did not appear that this procedure is being actively developed as a teratogen screening test. The extensive literature on both preimplantation and postimplantation embryo culture suggested considerable use of these two methods in evaluating embryotoxicants. The following discussion was compiled from information gleaned from all references. However, in the interest of brevity, only representative articles are specifically cited. Because the background and methodology for each system are distinct, each system will be discussed separately. PMID:3304996

  4. Long-term maintenance of human induced pluripotent stem cells by automated cell culture system.

    PubMed

    Konagaya, Shuhei; Ando, Takeshi; Yamauchi, Toshiaki; Suemori, Hirofumi; Iwata, Hiroo

    2015-01-01

    Pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem (iPS) cells, are regarded as new sources for cell replacement therapy. These cells can unlimitedly expand under undifferentiated conditions and be differentiated into multiple cell types. Automated culture systems enable the large-scale production of cells. In addition to reducing the time and effort of researchers, an automated culture system improves the reproducibility of cell cultures. In the present study, we newly designed a fully automated cell culture system for human iPS maintenance. Using an automated culture system, hiPS cells maintained their undifferentiated state for 60 days. Automatically prepared hiPS cells had a potency of differentiation into three germ layer cells including dopaminergic neurons and pancreatic cells. PMID:26573336

  5. An Easy-to-Use Polystyrene Microchip-based Cell Culture System.

    PubMed

    Tazawa, Hidekatsu; Sunaoshi, Shohei; Tokeshi, Manabu; Kitamori, Takehiko; Ohtani-Kaneko, Ritsuko

    2016-01-01

    In this study, we developed an integrated, low-cost microfluidic cell culture system that is easy to use. This system consists of a disposable polystyrene microchip, a polytetrafluoroethylene valve, an air bubble trap, and an indium tin oxide temperature controller. Valve pressure resistance was validated with a manometer to be 3 MPa. The trap protected against bubble contamination. The temperature controller enabled the culture of Macaca mulatta RF/6A 135 vascular endothelial cells, which are difficult to culture in glass microchips, without a CO2 incubator. We determined the optimal coating conditions for these cells and were able to achieve stable, confluent culture within 1 week. This practical system is suitable for low-cost screening and has potential applications as circulatory cell culture systems and research platforms in cell biology. PMID:26960617

  6. A Single-Cell and Feeder-Free Culture System for Monkey Embryonic Stem Cells

    PubMed Central

    Ono, Takashi; Suzuki, Yutaka; Kato, Yosuke; Fujita, Risako; Araki, Toshihiro; Yamashita, Tomoko; Kato, Hidemasa; Torii, Ryuzo; Sato, Naoya

    2014-01-01

    Primate pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), hold great potential for research and application in regenerative medicine and drug discovery. To maximize primate PSC potential, a practical system is required for generating desired functional cells and reproducible differentiation techniques. Much progress regarding their culture systems has been reported to date; however, better methods would still be required for their practical use, particularly in industrial and clinical fields. Here we report a new single-cell and feeder-free culture system for primate PSCs, the key feature of which is an originally formulated serum-free medium containing FGF and activin. In this culture system, cynomolgus monkey ESCs can be passaged many times by single-cell dissociation with traditional trypsin treatment and can be propagated with a high proliferation rate as a monolayer without any feeder cells; further, typical PSC properties and genomic stability can be retained. In addition, it has been demonstrated that monkey ESCs maintained in the culture system can be used for various experiments such as in vitro differentiation and gene manipulation. Thus, compared with the conventional culture system, monkey ESCs grown in the aforementioned culture system can serve as a cell source with the following practical advantages: simple, stable, and easy cell maintenance; gene manipulation; cryopreservation; and desired differentiation. We propose that this culture system can serve as a reliable platform to prepare primate PSCs useful for future research and application. PMID:24505480

  7. Biomaterials in co-culture systems: towards optimizing tissue integration and cell signaling within scaffolds.

    PubMed

    Battiston, Kyle G; Cheung, Jane W C; Jain, Devika; Santerre, J Paul

    2014-05-01

    Most natural tissues consist of multi-cellular systems made up of two or more cell types. However, some of these tissues may not regenerate themselves following tissue injury or disease without some form of intervention, such as from the use of tissue engineered constructs. Recent studies have increasingly used co-cultures in tissue engineering applications as these systems better model the natural tissues, both physically and biologically. This review aims to identify the challenges of using co-culture systems and to highlight different approaches with respect to the use of biomaterials in the use of such systems. The application of co-culture systems to stimulate a desired biological response and examples of studies within particular tissue engineering disciplines are summarized. A description of different analytical co-culture systems is also discussed and the role of biomaterials in the future of co-culture research are elaborated on. Understanding the complex cell-cell and cell-biomaterial interactions involved in co-culture systems will ultimately lead the field towards biomaterial concepts and designs with specific biochemical, electrical, and mechanical characteristics that are tailored towards the needs of distinct co-culture systems. PMID:24602569

  8. Algal culture studies related to a Closed Ecological Life Support System (CELSS)

    NASA Technical Reports Server (NTRS)

    Radmer, R.; Behrens, P.; Fernandez, E.; Ollinger, O.; Howell, C.; Venables, A.; Huggins, D.; Gladue, R.

    1984-01-01

    In many respects, algae would be the ideal plant component for a biologically based controlled life support system, since they are eminently suited to the closely coupled functions of atmosphere regeneration and food production. Scenedesmus obliquus and Spirulina platensis were grown in three continuous culture apparatuses. Culture vessels their operation and relative merits are described. Both light and nitrogen utilization efficiency are examined. Long term culture issues are detailed and a discussion of a plasmid search in Spirulina is included.

  9. Higher recovery rate of microorganisms from cerebrospinal fluid samples by the BACTEC culture system in comparison with agar culture.

    PubMed

    Calderaro, Adriana; Martinelli, Monica; Montecchini, Sara; Motta, Federica; Covan, Silvia; Larini, Sandra; Medici, Maria Cristina; Arcangeletti, Maria Cristina; Chezzi, Carlo; De Conto, Flora

    2016-04-01

    The aim of this study was to assess the diagnostic value of the BACTEC FX blood culture (BC) system as compared to the agar culture (AC) of cerebrospinal fluid samples (CSF), evaluating the recovery rate and the time to detection of microorganisms in a 3.5-year period. From December 2011 to May 2015, 1326 CSF samples (694 patients) were submitted to both AC and BC. Among the 150 positive samples (96 patients), 165 microorganisms were detected: 81 by both the protocols, 77 by BC alone, and 7 by AC alone, demonstrating a higher detection rate of BC (95.8%) than AC (53.3%). Although BC presents some disadvantages, it is able to improve the yield of clinically significant microorganisms, and it could potentially reduce the reporting time as compared to AC. The results obtained highlighted the necessity of a combined approach for the successful detection of central nervous system microbial infections. PMID:26867963

  10. Non-culturable bioaerosols in indoor settings: Impact on health and molecular approaches for detection

    NASA Astrophysics Data System (ADS)

    Blais-Lecours, Pascale; Perrott, Phillipa; Duchaine, Caroline

    2015-06-01

    Despite their significant impact on respiratory health, bioaerosols in indoor settings remain understudied and misunderstood. Culture techniques, predominantly used for bioaerosol characterisation in the past, allow for the recovery of only a small fraction of the real airborne microbial burden in indoor settings, given the inability of several microorganisms to grow on agar plates. However, with the development of new tools to detect non-culturable environmental microorganisms, the study of bioaerosols has advanced significantly. Most importantly, these techniques have revealed a more complex bioaerosol burden that also includes non-culturable microorganisms, such as archaea and viruses. Nevertheless, air quality specialists and consultants remain reluctant to adopt these new research-developed techniques, given that there are relatively few studies found in the literature, making it difficult to find a point of comparison. Furthermore, it is unclear as to how this new non-culturable data can be used to assess the impact of bioaerosol exposure on human health. This article reviews the literature that describes the non-culturable fraction of bioaerosols, focussing on bacteria, archaea and viruses, and examines its impact on bioaerosol-related diseases. It also outlines available molecular tools for the detection and quantification of these microorganisms and states various research needs in this field.

  11. Aerobic culture of methanogenic archaea without an external source of hydrogen.

    PubMed

    Khelaifia, S; Lagier, J-C; Nkamga, V D; Guilhot, E; Drancourt, M; Raoult, D

    2016-06-01

    Culturing methanogenic archaea is fastidious, expensive, and requires an external source of hydrogen and carbon dioxide. Until now, these microorganisms have only been cultivated under strictly anaerobic conditions. We previously developed a single versatile culture medium containing sugars and anti-oxydants for cultivating all human known methanogens. Performing aerobic cultures in the presence of Bacteroides thetaiotaomicron, which produces hydrogen, allows for cultivation of Methanobrevibacter smithii which itself produces methane. To obtain colonies, we cultivated M. smithii in an agar plate in the upper part of a double chamber flask with a liquid culture of B. thetaiotaomicron in the lower compartment. We subsequently cultured four other methanogenic species for the first time and successfully isolated 13 strains of M. smithii and nine strains of Methanobrevibacter oralis from 100 stools and 45 oral samples. This procedure allows aerobic isolation and antibiotic susceptibility testing. This changes the ability to routinely study methanogens, which have been neglected in clinical microbiology laboratories and may be useful for biogas production. PMID:27010812

  12. Design and Evaluation of a Cross-Cultural Training System

    NASA Technical Reports Server (NTRS)

    Santarelli, Thomas; Stagl, Kevin C.

    2011-01-01

    Cross-cultural competency, and the underlying communication and affective skills required to develop such expertise, is becoming increasingly important for a wide variety of domains. To address this need, we developed a blended learning platform which combines virtual role-play with tutorials, assessment and feedback. A Middle-Eastern Curriculum (MEC) exemplar for cross-cultural training U.S. military personnel was developed to guide the refinement of an existing game-based training platform. To complement this curriculum, we developed scenario authoring tools to enable end-users to define training objectives, link performance measures and feedback/remediation to these objectives, and deploy experiential scenarios within a game-based virtual environment (VE). Lessons learned from the design and development of this exemplar cross-cultural competency curriculum, as well as formative evaluation results, are discussed. Initial findings suggest that the underlying training technology promotes deep levels of semantic processing of the key information of relevant cultural and communication skills.

  13. Multi-Cultural Education: New Perspectives - New Delivery Systems.

    ERIC Educational Resources Information Center

    Dick, James; Van Every, Ivalyn J.

    This paper describes the cooperative development of multi-cultural inservice courses by the University of Nebraska Teacher Corps and the Omaha public schools. A planning group with representatives from the university and from the local school district met to develop courses to meet the specific needs of teachers in the project schools which…

  14. Bridging the Two Cultures: Disciplinary Divides and Educational Reward Systems

    ERIC Educational Resources Information Center

    Schiferl, E. I.

    2007-01-01

    In 1959 C.P. Snow believed that communication and education could span the cultural gap between the sciences and the humanities. In the twenty-first century, language, research models, and academic structures hinder intellectual communication between art history, cognitive neuroscience and perceptual psychology--three disciplines dedicated to…

  15. A Systemic Approach to Culturally Responsive Assessment Practices and Evaluation

    ERIC Educational Resources Information Center

    Slee, June

    2010-01-01

    In an earlier paper, Slee and Keenan demonstrated that it was possible for tertiary education institutions to design culturally responsive assessment procedures that complied with standardised assessment policy. The authors' paper described "Growing Our Own," an initiative between Charles Darwin University and Northern Territory Catholic…

  16. Modular plant culture systems for life support functions

    NASA Technical Reports Server (NTRS)

    1985-01-01

    The current state of knowledge with regard to culture of higher plants in the zero-G environment is assessed; and concepts for the empirical development of small plant growth chambers for the production of salad type vegetables on space shuttle or space station are evaluated. American and Soviet space flight experiences in gravitational biology are summarized.

  17. Educational Policy vs. Culturally Sensitive Programs in Turkish Educational System

    ERIC Educational Resources Information Center

    Arslan, Hasan

    2009-01-01

    The purpose of this study is to examine the perceptions of elementary school teachers about the sensitiveness of principals, teachers, and curriculum on multicultural education. Education provides the transmission and the advancement of its culture while it is developing and enhancing the common values, the integrity and the progress of…

  18. Algal culture studies related to a Closed Ecological Life Support System (CELSS)

    NASA Technical Reports Server (NTRS)

    Radmer, R. O.; Ollinger, O.; Venables, A.; Fernandez, E.

    1982-01-01

    Studies with algal cultures which relate to closed ecological life support systems (CELSS) are discussed. A description of a constant cell density apparatus for continuous culture of algae is included. Excretion of algal by-products, and nitrogen utilization and excretion are discussed.

  19. MOSAICA: A Web-2.0 Based System for the Preservation and Presentation of Cultural Heritage

    ERIC Educational Resources Information Center

    Barak, Miri; Herscoviz, Orit; Kaberman, Zvia; Dori, Yehudit J.

    2009-01-01

    The question of how to present cultural heritage resources in a way that attracts potential users is becoming important in our ever-changing world. This paper describes MOSAICA system--a web 2.0-based toolbox, dedicated for the preservation and presentation of cultural heritage. This paper also describes an evaluation study that examined MOSAICA…

  20. The Relationship between National Culture and the Usability of an E-Learning System

    ERIC Educational Resources Information Center

    Adeoye, Blessing; Wentling, Rose Mary

    2007-01-01

    The purpose of this study was to investigate possible relationships between national culture and the usability of an e-learning system. The theoretical frameworks that were used to guide this study were Hofstede's (1980) cultural dimensions, and Nielson's (1993) usability attributes. The sample for this study was composed of 24 international…

  1. Multiple Embedded Inequalities and Cultural Diversity in Educational Systems: A Theoretical and Empirical Exploration

    ERIC Educational Resources Information Center

    Verhoeven, Marie

    2011-01-01

    This article explores the social construction of cultural diversity in education, with a view to social justice. It examines how educational systems organize ethno-cultural difference and how this process contributes to inequalities. Theoretical resources are drawn from social philosophy as well as from recent developments in social organisation…

  2. Revisiting the Role of Cultural Capital in East Asian Educational Systems: The Case of South Korea

    PubMed Central

    Byun, Soo-yong; Schofer, Evan; Kim, Kyung-keun

    2013-01-01

    The concept of cultural capital has proved invaluable in understanding educational systems in Western countries, and recent work seeks to extend those insights to the diverse educational systems of other geographic regions. We explored cultural capital in South Korea by investigating the relationships among family socioeconomic status (SES), cultural capital, and children's academic achievement using data from the 2000 Programme for International Student Assessment. South Korea was compared with Japan, France, and the United States to understand how institutional features of South Korean education shape the role of cultural capital in academic success. Results showed that family SES had a positive effect on both parental objectified cultural capital and children's embodied cultural capital in South Korea, consistent with evidence from the other countries. Moreover, parental objectified cultural capital had a positive effect on children's academic achievement in South Korea. In contrast to other countries, however, children's embodied cultural capital had a negative effect on academic achievement in South Korea controlling for the other variables. We highlighted several institutional features of South Korean education including a standardized curriculum, extreme focus on test preparation, and extensive shadow education, which may combine to suppress the effect of children's embodied cultural capital on academic achievement. PMID:24285909

  3. Micro 3D cell culture systems for cellular behavior studies: Culture matrices, devices, substrates, and in-situ sensing methods.

    PubMed

    Choi, Jonghoon; Lee, Eun Kyu; Choo, Jaebum; Yuh, Junhan; Hong, Jong Wook

    2015-09-01

    Microfabricated systems equipped with 3D cell culture devices and in-situ cellular biosensing tools can be a powerful bionanotechnology platform to investigate a variety of biomedical applications. Various construction substrates such as plastics, glass, and paper are used for microstructures. When selecting a construction substrate, a key consideration is a porous microenvironment that allows for spheroid growth and mimics the extracellular matrix (ECM) of cell aggregates. Various bio-functionalized hydrogels are ideal candidates that mimic the natural ECM for 3D cell culture. When selecting an optimal and appropriate microfabrication method, both the intended use of the system and the characteristics and restrictions of the target cells should be carefully considered. For highly sensitive and near-cell surface detection of excreted cellular compounds, SERS-based microsystems capable of dual modal imaging have the potential to be powerful tools; however, the development of optical reporters and nanoprobes remains a key challenge. We expect that the microsystems capable of both 3D cell culture and cellular response monitoring would serve as excellent tools to provide fundamental cellular behavior information for various biomedical applications such as metastasis, wound healing, high throughput screening, tissue engineering, regenerative medicine, and drug discovery and development. PMID:26358782

  4. A Well-Controlled Nucleus Pulposus Tissue Culture System with Injection Port for Evaluating Regenerative Therapies.

    PubMed

    Arkesteijn, Irene T M; Mouser, Vivian H M; Mwale, Fackson; van Dijk, Bart G M; Ito, Keita

    2016-05-01

    In vitro evaluation of nucleus pulposus (NP) tissue regeneration would be useful, but current systems for NP culture are not ideal for injections. The aim of this study was to develop a long-term culture system for NP tissue that allows injections of regenerative agents. Bovine caudal NPs were harvested and placed in the newly designed culture system. After equilibration of the tissue to 0.3 MPa the volume was fixed and the tissue was cultured for 28 days. The cell viability and extracellular matrix composition remained unchanged during the culture period and gene expression profiles were similar to those obtained in earlier studies. Furthermore, to test the responsiveness of bovine caudal NPs in the system, samples were cultured for 4 days and injected twice (day 1 and 3) with (1) PBS, (2) Link-N, for regeneration, and (3) TNF-α, for degeneration. It was shown that TNF-α increased COX2 gene expression, whereas no effect of Link-N was detected. In conclusion, the newly designed system allows long-term culture of NP tissue, wherein tissue reactions to injected stimulants can be observed. PMID:26294008

  5. Medical Devices; Obstetrical and Gynecological Devices; Classification of the Intravaginal Culture System. Final order.

    PubMed

    2016-01-01

    The Food and Drug Administration (FDA) is classifying the intravaginal culture system into class II (special controls). The special controls that will apply to the device are identified in this order and will be part of the codified language for the intravaginal culture system's classification. The Agency is classifying the device into class II (special controls) in order to provide a reasonable assurance of safety and effectiveness of the device. PMID:26742184

  6. In Vitro Culture of Previously Uncultured Oral Bacterial Phylotypes

    PubMed Central

    Thompson, Hayley; Rybalka, Alexandra; Moazzez, Rebecca; Dewhirst, Floyd E.

    2015-01-01

    Around a third of oral bacteria cannot be grown using conventional bacteriological culture media. Community profiling targeting 16S rRNA and shotgun metagenomics methods have proved valuable in revealing the complexity of the oral bacterial community. Studies investigating the role of oral bacteria in health and disease require phenotypic characterizations that are possible only with live cultures. The aim of this study was to develop novel culture media and use an in vitro biofilm model to culture previously uncultured oral bacteria. Subgingival plaque samples collected from subjects with periodontitis were cultured on complex mucin-containing agar plates supplemented with proteose peptone (PPA), beef extract (BEA), or Gelysate (GA) as well as on fastidious anaerobe agar plus 5% horse blood (FAA). In vitro biofilms inoculated with the subgingival plaque samples and proteose peptone broth (PPB) as the growth medium were established using the Calgary biofilm device. Specific PCR primers were designed and validated for the previously uncultivated oral taxa Bacteroidetes bacteria HOT 365 and HOT 281, Lachnospiraceae bacteria HOT 100 and HOT 500, and Clostridiales bacterium HOT 093. All agar media were able to support the growth of 10 reference strains of oral bacteria. One previously uncultivated phylotype, Actinomyces sp. HOT 525, was cultivated on FAA. Of 93 previously uncultivated phylotypes found in the inocula, 26 were detected in in vitro-cultivated biofilms. Lachnospiraceae bacterium HOT 500 was successfully cultured from biofilm material harvested from PPA plates in coculture with Parvimonas micra or Veillonella dispar/parvula after colony hybridization-directed enrichment. The establishment of in vitro biofilms from oral inocula enables the cultivation of previously uncultured oral bacteria and provides source material for isolation in coculture. PMID:26407883

  7. The Role of State Early Childhood Comprehensive Systems in Promoting Cultural Competence and Effective Cross-Cultural Communication. Building State Early Childhood Comprehensive Systems Series, Number 8

    ERIC Educational Resources Information Center

    Sareen, Harvinder; Visencio, Diane; Russ, Shirley; Halfon, Neal

    2005-01-01

    If early childhood systems are to be effective at the population level then they must be able to provide family-centered care to all the racial, ethnic and cultural groups that they serve. Despite major policy driven and technological advances in healthcare, health disparities across different races and ethnicities persist. For example, the infant…

  8. [Effects of different culture system of isolating and passage of sheep embryonic stem-like cells].

    PubMed

    Bai, Changming; Liu, Chousheng; Wang, Zhigang; Wang, Xinzhuang

    2008-07-01

    In this research, we use mouse embryonic fibroblasts as feeder layers. To eliminate the influence of serum and mouse embryonic stem cells (ESCs) conditioned medium (ESCCM) on self-renewal of sheep embryonic stem-like cells, knockout serum replacement (KSR) was used to replace serum, then supplanted with ESCCM for the isolation and cloning of sheep embryonic stem-like cells. We found when inner cell masses (ICMs) cultured in the control group with medium supplanted with fetal bovine serum (FBS), sheep ES-like cells could not survive for more than 3 passages. However, sheep embryonic stem-like cells could remain undifferentiated for 5 passages when cultured in the medium that FBS was substituted by KSR. The result indicates that KSR culture system was more suitable for the isolation and cloning of sheep embryonic stem-like cells compared to FBS culture system. Finally we applied medium with 15% KSR as basic medium supplanted with 40% ESCCM as a new culture system to isolate sheep embryonic stem-like cells, we found one embryonic stem-like cell line still maintained undifferentiating for 8 passages, which characterized with a normal and stable karyotype and high expression of alkaline phosphatase. These results suggest that it is suitable to culture sheep ICM in the new culture system with 15% KSR as basic medium and supplanted with 40% ESCCM, which indicated that mouse ES cells might secrete factors playing important roles in promoting sheep ES-like cells' self-renewal. PMID:18837407

  9. The Changeable Nervous System: Studies On Neuroplasticity In Cerebellar Cultures

    PubMed Central

    Seil, Fredrick J.

    2014-01-01

    Circuit reorganization after injury was studied in a cerebellar culture model. When cerebellar cultures derived from newborn mice were exposed at explantation to a preparation of cytosine arabinoside that destroyed granule cells and oligodendrocytes and compromised astrocytes, Purkinje cells surviving in greater than usual numbers were unensheathed by astrocytic processes and received twice the control number of inhibitory axosomatic synapses. Purkinje cell axon collaterals sprouted and many of their terminals formed heterotypical synapses with other Purkinje cell dendritic spines. The resulting circuit reorganization preserved inhibition in the cerebellar cortex. Following this reorganization, replacement of the missing granule cells and glia was followed by a restitution of the normal circuitry. Most of these developmental and reconstructive changes were not dependent on neuronal activity, the major exception being inhibitory synaptogenesis. The full complement of inhibitory synapses did not develop in the absence of neuronal activity, which could be mitigated by application of exogenous TrkB receptor ligands. Inhibitory synaptogenesis could also be promoted by activity-induced release of endogenous TrkB receptor ligands or by antibody activation of the TrkB receptor. PMID:24933693

  10. Lysine hydroxylation of collagen in a fibroblast cell culture system

    NASA Technical Reports Server (NTRS)

    Uzawa, Katsuhiro; Yeowell, Heather N.; Yamamoto, Kazushi; Mochida, Yoshiyuki; Tanzawa, Hideki; Yamauchi, Mitsuo

    2003-01-01

    The lysine (Lys) hydroxylation pattern of type I collagen produced by human fibroblasts in culture was analyzed and compared. Fibroblasts were cultured from normal human skin (NSF), keloid (KDF), fetal skin (FDF), and skin tissues of Ehlers-Danlos syndrome type VIA and VIB patients (EDS-VIA and -VIB). The type I collagen alpha chains with or without non-helical telopeptides were purified from the insoluble matrix and analyzed. In comparison with NSFs, KDF and FDF showed significantly higher Lys hydroxylation, particularly in the telopeptide domains of both alpha chains. Both EDS-VIA and -VIB showed markedly lower Lys hydroxylation in the helical domains of both alpha chains whereas that in the telopeptides was comparable with those of NSFs. A similar profile was observed in the tissue sample of the EDS-VIB patient. These results demonstrate that the Lys hydroxylation pattern is domain-specific within the collagen molecule and that this method is useful to characterize the cell phenotypes in normal/pathological connective tissues.

  11. How Many Samples and How Many Culture Media To Diagnose a Prosthetic Joint Infection: a Clinical and Microbiological Prospective Multicenter Study

    PubMed Central

    Léger, Julie; Tandé, Didier; Plouzeau, Chloé; Valentin, Anne Sophie; Jolivet-Gougeon, Anne; Lemarié, Carole; Kempf, Marie; Héry-Arnaud, Geneviève; Bret, Laurent; Juvin, Marie Emmanuelle; Giraudeau, Bruno; Burucoa, Christophe

    2015-01-01

    Although numerous perioperative samples and culture media are required to diagnose prosthetic joint infection (PJI), their exact number and types have not yet been definitely determined with a high level of proof. We conducted a prospective multicenter study to determine the minimal number of samples and culture media required for accurate diagnosis of PJI. Over a 2-year period, consecutive patients with clinical signs suggesting PJI were included, with five perioperative samples per patient. The bacteriological and PJI diagnosis criteria were assessed using a random selection of two, three, or four samples and compared with those obtained using the recommended five samples (references guidelines). The results obtained with two or three culture media were then compared with those obtained with five culture media for both criteria. The times-to-positivity of the different culture media were calculated. PJI was confirmed in 215/264 suspected cases, with a bacteriological criterion in 192 (89%). The PJI was monomicrobial (85%) or polymicrobial (15%). Percentages of agreement of 98.1% and 99.7%, respectively, for the bacteriological criterion and confirmed PJI diagnosis were obtained when four perioperative samples were considered. The highest percentages of agreement were obtained with the association of three culture media, a blood culture bottle, a chocolate agar plate, and Schaedler broth, incubated for 5, 7, and 14 days, respectively. This new procedure leads to significant cost saving. Our prospective multicenter study showed that four samples seeded on three culture media are sufficient for diagnosing PJI. PMID:26637380

  12. How Many Samples and How Many Culture Media To Diagnose a Prosthetic Joint Infection: a Clinical and Microbiological Prospective Multicenter Study.

    PubMed

    Bémer, Pascale; Léger, Julie; Tandé, Didier; Plouzeau, Chloé; Valentin, Anne Sophie; Jolivet-Gougeon, Anne; Lemarié, Carole; Kempf, Marie; Héry-Arnaud, Geneviève; Bret, Laurent; Juvin, Marie Emmanuelle; Giraudeau, Bruno; Corvec, Stéphane; Burucoa, Christophe

    2016-02-01

    Although numerous perioperative samples and culture media are required to diagnose prosthetic joint infection (PJI), their exact number and types have not yet been definitely determined with a high level of proof. We conducted a prospective multicenter study to determine the minimal number of samples and culture media required for accurate diagnosis of PJI. Over a 2-year period, consecutive patients with clinical signs suggesting PJI were included, with five perioperative samples per patient. The bacteriological and PJI diagnosis criteria were assessed using a random selection of two, three, or four samples and compared with those obtained using the recommended five samples (references guidelines). The results obtained with two or three culture media were then compared with those obtained with five culture media for both criteria. The times-to-positivity of the different culture media were calculated. PJI was confirmed in 215/264 suspected cases, with a bacteriological criterion in 192 (89%). The PJI was monomicrobial (85%) or polymicrobial (15%). Percentages of agreement of 98.1% and 99.7%, respectively, for the bacteriological criterion and confirmed PJI diagnosis were obtained when four perioperative samples were considered. The highest percentages of agreement were obtained with the association of three culture media, a blood culture bottle, a chocolate agar plate, and Schaedler broth, incubated for 5, 7, and 14 days, respectively. This new procedure leads to significant cost saving. Our prospective multicenter study showed that four samples seeded on three culture media are sufficient for diagnosing PJI. PMID:26637380

  13. Cells and Culture Systems Used to Model the Small Airway Epithelium.

    PubMed

    Bhowmick, Rudra; Gappa-Fahlenkamp, Heather

    2016-06-01

    The pulmonary epithelium is divided into upper, lower, and alveolar (or small) airway epithelia and acts as the mechanical and immunological barrier between the external environment and the underlying submucosa. Of these, the small airway epithelium is the principal area of gas exchange and has high immunological activity, making it a major area of cell biology, immunology, and pharmaceutical research. As animal models do not faithfully represent the human pulmonary system and ex vivo human lung samples have reliability and availability issues, cell lines, and primary cells are widely used as small airway epithelial models. In vitro, these cells are mostly cultured as monolayers (2-dimensional cultures), either media submerged or at air-liquid interface. However, these 2-dimensional cultures lack a three dimension-a scaffolding extracellular matrix, which establishes the intercellular network in the in vivo airway epithelium. Therefore, 3-dimensional cell culture is currently a major area of development, where cells are cultured in a matrix or are cultured in a manner that they develop ECM-like scaffolds between them, thus mimicking the in vivo phenotype more faithfully. This review focuses on the commonly used small airway epithelial cells, their 2-dimensional and 3-dimensional culture techniques, and their comparative phenotype when cultured under these systems. PMID:27071933

  14. Evaluation of a new biphasic culture system for the recovery of mycobacteria.

    PubMed

    Giger, T; Burkardt, H J

    1990-06-01

    A newly developed biphasic culture system (MB-Check) for recovery of mycobacteria was evaluated. The biphasic system consists of a bottle containing selective modified Middlebrook 7H9 broth and a mounted dip slide with chocolate agar and modified Middlebrook 7H11 agar with and without NAP. The system was compared with culture on two egg-based media, Lowenstein medium and a selective Gottsacker medium, using 995 routine specimens and 90 artificially seeded sputa. Mycobacterium tuberculosis was detected in 17 of the 995 routine specimens by the biphasic system and in 14 specimens by the egg-based media together. In the artificially seeded sputa the biphasic system showed higher sensitivity in detection of both tuberculosis complex and non-tuberculous mycobacteria than the egg-based media. The recovery times of the new system were comparable to those of the two conventional culture methods. PMID:2387296

  15. Unique cell culture systems for ground based research

    NASA Technical Reports Server (NTRS)

    Lewis, Marian L.

    1990-01-01

    The horizontally rotating fluid-filled, membrane oxygenated bioreactors developed at NASA Johnson for spacecraft applications provide a powerful tool for ground-based research. Three-dimensional aggregates formed by cells cultured on microcarrier beads are useful for study of cell-cell interactions and tissue development. By comparing electron micrographs of plant seedlings germinated during Shuttle flight 61-C and in an earth-based rotating bioreactor it is shown that some effects of microgravity are mimicked. Bioreactors used in the UAH Bioreactor Laboratory will make it possible to determine some of the effects of altered gravity at the cellular level. Bioreactors can be valuable for performing critical, preliminary-to-spaceflight experiments as well as medical investigations such as in vitro tumor cell growth and chemotherapeutic drug response; the enrichment of stem cells from bone marrow; and the effect of altered gravity on bone and muscle cell growth and function and immune response depression.

  16. Evaluation of the PREVI® Isola automated seeder system compared to reference manual inoculation for antibiotic susceptibility testing by the disk diffusion method.

    PubMed

    Le Page, S; van Belkum, A; Fulchiron, C; Huguet, R; Raoult, D; Rolain, J-M

    2015-09-01

    The disk diffusion (DD) method remains the most popular manual technique for antibiotic susceptibility testing (AST) in clinical microbiology laboratories. This is because of its simplicity, reproducibility, and limited cost compared to (automated) microdilution systems, which are usually less sensitive at detecting certain important mechanisms of resistance. Here, we evaluate the PREVI® Isola automated seeder system using a new protocol for spreading bacterial suspensions (eight deposits of calibrated inocula of bacteria, followed by two rounds of rotation) in comparison with manual DD reference testing on a large series of clinical and reference strains. The average time required for seeding one agar plate for DD with this new protocol was 51 s per plate, i.e., 70 agar plates/h. Reproducibility and repeatability was assessed on three reference and three randomly chosen clinical strains, as usually requested by the European Committee on Antimicrobial Susceptibility Testing (EUCAST), and was excellent compared to the manual method. The standard deviations of zones of growth inhibition showed no statistical discrimination. The correlation between the two methods, assessed using 294 clinical isolates and a panel of six antibiotics (n = 3,528 zones of growth inhibition measured), was excellent, with a correlation coefficient of 0.977. The new PREVI® Isola protocol adapted for DD had a sensitivity of 99 % and a specificity of 100 % compared to the manual technique for interpreting DD as recommended by the EUCAST. PMID:26092031

  17. Ripples in a pond: an open system model of the evolution of safety culture.

    PubMed

    Morley, F J Joel; Harris, Don

    2006-01-01

    The development of an effective safety culture is essential to promote safe operations. Previous studies have either identified the characteristics of effective safety culture analytically, inferring them from signs and symbols derived from working practices, or have restricted the study of the development of safety culture to workers within an organisation. This paper describes a large-scale survey-based study in which the factors influencing the evolution of safety culture are identified empirically and, drawing upon open systems theory, are also extended beyond the bounds of the organisation. Three major determinants of safety culture are identified: safety concerns, influences and actions. Sub-components within each of these categories are also identified and the relationship between them is hypothesised. PMID:16553996

  18. In Vitro Cultivation of ‘Unculturable’ Oral Bacteria, Facilitated by Community Culture and Media Supplementation with Siderophores

    PubMed Central

    Vartoukian, Sonia R.; Adamowska, Aleksandra; Lawlor, Megan; Moazzez, Rebecca; Dewhirst, Floyd E.; Wade, William G.

    2016-01-01

    Over a third of oral bacteria are as-yet-uncultivated in-vitro. Siderophores have been previously shown to enable in-vitro growth of previously uncultivated bacteria. The objective of this study was to cultivate novel oral bacteria in siderophore-supplemented culture media. Various compounds with siderophore activity, including pyoverdines-Fe-complex, desferricoprogen and salicylic acid, were found to stimulate the growth of difficult-to-culture strains Prevotella sp. HOT-376 and Fretibacterium fastidiosum. Furthermore, pyrosequencing analysis demonstrated increased proportions of the as-yet-uncultivated phylotypes Dialister sp. HOT-119 and Megasphaera sp. HOT-123 on mixed culture plates supplemented with siderophores. Therefore a culture model was developed, which incorporated 15 μg siderophore (pyoverdines-Fe-complex or desferricoprogen) or 150 μl neat subgingival-plaque suspension into a central well on agar plates that were inoculated with heavily-diluted subgingival-plaque samples from subjects with periodontitis. Colonies showing satellitism were passaged onto fresh plates in co-culture with selected helper strains. Five novel strains, representatives of three previously-uncultivated taxa (Anaerolineae bacterium HOT-439, the first oral taxon from the Chloroflexi phylum to have been cultivated; Bacteroidetes bacterium HOT-365; and Peptostreptococcaceae bacterium HOT-091) were successfully isolated. All novel isolates required helper strains for growth, implying dependence on a biofilm lifestyle. Their characterisation will further our understanding of the human oral microbiome. PMID:26764907

  19. Tissue culture system using a PANDA ring resonator and wavelength router for hydroponic plant.

    PubMed

    Kamoldilok, Surachart; Suwanpayak, Nathaporn; Suttirak, Saisudawan; Yupapin, Preecha P

    2012-06-01

    A novel system of nanofluidics trapping and delivery, which is known as a tissue culture system is proposed. By using the intense optical pulse(i.e., a soliton pulse) and a system constructed by a liquid core waveguide, the optical vortices (gradient optical fields/wells) can be generated, where the trapping tools in the same way as the optical tweezers in the PANDA ring resonator can be formed. By controlling the suitable parameters, the intense optical vortices can be generated within the PANDA ring resonator, in which the nanofluidics can be trapped and moved (transported) dynamically within the Tissue culture system(a wavelength router), which can be used for tissue culture and delivery in the hydroponic plant system. PMID:22411055

  20. Changing the Management Culture in a Public School System

    ERIC Educational Resources Information Center

    Cohen, Allan R.; Gadon, Herman

    1978-01-01

    The intervention described led to the creation of a new management structure for a public school system with different decision-making processes, greater administrator involvement, and increased commitment to the total system. A number of propositions for change agents have been formulated. Available from: JABS Order Dept., NTL Institute for…

  1. Investigating Educational Systems, Leadership, and School Culture: A Holistic Approach

    ERIC Educational Resources Information Center

    Pratt, Jill Elizabeth

    2014-01-01

    Most populous school districts operate using a bureaucratic hierarchical organizational structure developed primarily for industry, a system structure that has remained intact for a century despite evolving from a manufacturing to a knowledge-based economy. Although strong for efficiency, this system structure is resistant to change and promotes…

  2. Development of 3D hydrogel culture systems with on-demand cell separation.

    PubMed

    Hamilton, Sharon K; Bloodworth, Nathaniel C; Massad, Christopher S; Hammoudi, Taymour M; Suri, Shalu; Yang, Peter J; Lu, Hang; Temenoff, Johnna S

    2013-04-01

    Recently there has been an increased interest in the effects of paracrine signaling between groups of cells, particularly in the context of better understanding how stem cells contribute to tissue repair. Most current 3D co-culture methods lack the ability to effectively separate two cell populations after the culture period, which is important for simultaneously analyzing the reciprocal effects of each cell type on the other. Here, we detail the development of a 3D hydrogel co-culture system that allows us to culture different cell types for up to 7 days and subsequently separate and isolate the different cell populations using enzyme-sensitive glues. Separable 3D co-culture laminates were prepared by laminating PEG-based hydrogels with enzyme-degradable hydrogel adhesives. Encapsulated cell populations exhibited good segregation with well-defined interfaces. Furthermore, constructs can be separated on-demand upon addition of the appropriate enzyme, while cell viability remains high throughout the culture period, even after laminate separation. This platform offers great potential for a variety of basic cell signaling studies as the incorporation of an enzyme-sensitive adhesive interface allows the on-demand separation of individual cell populations for immediate analysis or further culture to examine persistence of co-culture effects and paracrine signaling on cell populations. PMID:23447378

  3. Self-driven perfusion culture system using a paper-based double-layered scaffold.

    PubMed

    Ozaki, Ai; Arisaka, Yoshinori; Takeda, Naoya

    2016-01-01

    Shear stress caused by fluid flow is known to promote tissue development from cells in vivo. Therefore, perfusion cultures have been studied to investigate the mechanisms involved and to fabricate engineered tissues in vitro, particularly those that include blood vessels. Microfluidic devices, which function with fine machinery of chambers and microsyringes for fluid flow and have small culture areas, are conventionally used for perfusion culture. In contrast, we have developed a self-driven perfusion culture system by using a paper-based double-layered scaffold as the fundamental component. Gelatin microfibers were electrospun onto a paper material to prepare the scaffold system, in which the constant perfusion of the medium and the scaffold for cell adhesion/proliferation were functionally divided into a paper and a gelatin microfiber layer, respectively. By applying both the capillary action and siphon phenomenon of the paper-based scaffold, which bridged two medium chambers at different height levels, a self-driven medium flow was achieved and the flow rate was also stable, constant, and quantitatively controllable. Moreover, the culture area was enlargeable to the cm(2) scale. The endothelial cells cultivated on this system oriented along the medium-flow direction, suggesting that the shear stress caused by medium flow was effectively applied. This perfusion culture system is expected to be useful for fabricating three-dimensional and large engineered tissues in the future. PMID:27550929

  4. In Vivo Bone Regeneration Using Tubular Perfusion System Bioreactor Cultured Nanofibrous Scaffolds

    PubMed Central

    Yeatts, Andrew B.; Both, Sanne K.; Yang, Wanxun; Alghamdi, Hamdan S.; Yang, Fang; Jansen, John A.

    2014-01-01

    The use of bioreactors for the in vitro culture of constructs for bone tissue engineering has become prevalent as these systems may improve the growth and differentiation of a cultured cell population. Here we utilize a tubular perfusion system (TPS) bioreactor for the in vitro culture of human mesenchymal stem cells (hMSCs) and implant the cultured constructs into rat femoral condyle defects. Using nanofibrous electrospun poly(lactic-co-glycolic acid)/poly(ɛ-caprolactone) scaffolds, hMSCs were cultured for 10 days in vitro in the TPS bioreactor with cellular and acellular scaffolds cultured statically for 10 days as a control. After 3 and 6 weeks of in vivo culture, explants were removed and subjected to histomorphometric analysis. Results indicated more rapid bone regeneration in defects implanted with bioreactor cultured scaffolds with a new bone area of 1.23±0.35 mm2 at 21 days compared to 0.99±0.43 mm2 and 0.50±0.29 mm2 in defects implanted with statically cultured scaffolds and acellular scaffolds, respectively. At the 21 day timepoint, statistical differences (p<0.05) were only observed between defects implanted with cell containing scaffolds and the acellular control. After 42 days, however, defects implanted with TPS cultured scaffolds had the greatest new bone area with 1.72±0.40 mm2. Defects implanted with statically cultured and acellular scaffolds had a new bone area of 1.26±0.43 mm2 and 1.19±0.33 mm2, respectively. The increase in bone growth observed in defects implanted with TPS cultured scaffolds was statistically significant (p<0.05) when compared to both the static and acellular groups at this timepoint. This study demonstrates the efficacy of the TPS bioreactor to improve bone tissue regeneration and highlights the benefits of utilizing perfusion bioreactor systems to culture MSCs for bone tissue engineering. PMID:23865551

  5. Lab on a chip-based hepatic sinusoidal system simulator for optimal primary hepatocyte culture.

    PubMed

    Choi, Yoon Young; Kim, Jaehyung; Lee, Sang-Hoon; Kim, Dong-Sik

    2016-08-01

    Primary hepatocyte cultures have been used in studies on liver disease, physiology, and pharmacology. While they are an important tool for in vitro liver studies, maintaining liver-specific characteristics of hepatocytes in vitro is difficult, as these cells rapidly lose their unique characteristics and functions. Portal flow is an important condition to preserve primary hepatocyte functions and liver regeneration in vivo. We have developed a microfluidic chip that does not require bulky peripheral devices or an external power source to investigate the relationship between hepatocyte functional maintenance and flow rates. In our culture system, two types of microfluidic devices were used as scaffolds: a monolayer- and a concave chamber-based device. Under flow conditions, our chips improved albumin and urea secretion rates after 13 days compared to that of the static chips. Reverse transcription polymerase chain reaction demonstrated that hepatocyte-specific gene expression was significantly higher at 13 days under flow conditions than when using static chips. For both two-dimensional and three-dimensional culture on the chips, flow resulted in the best performance of the hepatocyte culture in vitro. We demonstrated that flow improves the viability and efficiency of long-term culture of primary hepatocytes and plays a key role in hepatocyte function. These results suggest that this flow system has the potential for long-term hepatocyte cultures as well as a technique for three-dimensional culture. PMID:27334878

  6. Glycoprotein secretion in a tracheal organ culture system

    SciTech Connect

    Warunek, D.J.

    1985-01-01

    Glycoprotein secretion in the rat trachea was studied in vitro, utilizing a modified, matrix embed/perfusion chamber. Baseline parameters of the culture environment were determined by enzymatic and biochemical procedures. The effect of pilocarpine on the release of labelled glycoproteins from the tracheal epithelium was assessed. After a single stimulation with the drug, there was a significant increase in the release of /sup 14/C-glucosamine and /sup 3/H-fucose-labelled glycoprotein. The response was dose-dependent. Similar results were obtained after a second exposure to pilocarpine. However, no dose response was observed. Morphological analyses of the tracheal epithelial secretory cells by Alcian Blue/Periodic Acid Schiff staining showed a significant decrease in the total number of Alcian Blue staining cells and an increase in the mixed cell population after a single exposure to pilocarpine. Second stimulation with the drug showed that the trachea was able to respond again, this time with a further decrease in the number of Alcian Blue staining cells and a decrease in the PAS staining cells as well. Carbohydrate analyses after the first simulation with pilocarpine showed increased levels of N-acetyl neuraminic acid and the neutral carbohydrates, fucose and galactose, in the precipitated glycoproteins.

  7. Dynamic cell culture system: a new cell cultivation instrument for biological experiments in space

    NASA Technical Reports Server (NTRS)

    Gmunder, F. K.; Nordau, C. G.; Tschopp, A.; Huber, B.; Cogoli, A.

    1988-01-01

    The prototype of a miniaturized cell cultivation instrument for animal cell culture experiments aboard Spacelab is presented (Dynamic cell culture system: DCCS). The cell chamber is completely filled and has a working volume of 200 microliters. Medium exchange is achieved with a self-powered osmotic pump (flowrate 1 microliter h-1). The reservoir volume of culture medium is 230 microliters. The system is neither mechanically stirred nor equipped with sensors. Hamster kidney (Hak) cells growing on Cytodex 3 microcarriers were used to test the biological performance of the DCCS. Growth characteristics in the DCCS, as judged by maximal cell density, glucose consumption, lactic acid secretion and pH, were similar to those in cell culture tubes.

  8. Magnetic field-magnetic nanoparticle culture system used to grow in vitro murine embryonic stem cells.

    PubMed

    de Freitas, Erika Regina Leal; Soares, Paula Roberta Otaviano; de Santos, Rachel Paula; dos Santos, Regiane Lopes; Porfírio, Elaine Paulucio; Báo, Sônia N; Lima, Emília Celma Oliveira; Guillo, Lídia Andreu

    2011-01-01

    The in vitro growth of embryonic stem cells (ESCs) is usually obtained in the presence of murine embryonic fibroblasts (MEF), but new methods for in vitro expansion of ESCs should be developed due to their potential clinical use. This study aims to establish a culture system to expand and maintain ESCs in the absence of MEF by using murine embryonic stem cells (mECS) as a model of embryonic stem cell. Magnetic nanoparticles (MNPs) were used for growing mESCs in the presence of an external magnetic field, creating the magnetic field-magnetic nanoparticle (MF-MNP) culture system. The growth characteristics were evaluated showing a doubling time slightly higher for mESCs cultivated in the presence of the system than in the presence of the MEF. The undifferentiated state was characterized by RT-PCR, immunofluorescence, alkaline phosphatase activity and electron microscopy. Murine embryonic stem cells cultivated in presence of the MF-MNP culture system exhibited Oct-4 and Nanog expression and high alkaline phosphatase activity. Ultrastructural morphology showed that the MF-MNP culture system did not interfere with processes that cause structural changes in the cytoplasm or nucleus. The MF-MNP culture system provides a tool for in vitro expansion of mESCs and could contribute to studies that aim the therapeutic use of embryonic stem cells. PMID:21446404

  9. System for measuring oxygen consumption rates of mammalian cells in static culture under hypoxic conditions.

    PubMed

    Kagawa, Yuki; Miyahara, Hirotaka; Ota, Yuri; Tsuneda, Satoshi

    2016-01-01

    Estimating the oxygen consumption rates (OCRs) of mammalian cells in hypoxic environments is essential for designing and developing a three-dimensional (3-D) cell culture system. However, OCR measurements under hypoxic conditions are infrequently reported in the literature. Here, we developed a system for measuring OCRs at low oxygen levels. The system injects nitrogen gas into the environment and measures the oxygen concentration by an optical oxygen microsensor that consumes no oxygen. The developed system was applied to HepG2 cells in static culture. Specifically, we measured the spatial profiles of the local dissolved oxygen concentration in the medium, then estimated the OCRs of the cells. The OCRs, and also the pericellular oxygen concentrations, decreased nonlinearly as the oxygen partial pressure in the environment decreased from 19% to 1%. The OCRs also depended on the culture period and the matrix used for coating the dish surface. Using this system, we can precisely estimate the OCRs of various cell types under environments that mimic 3-D culture conditions, contributing crucial data for an efficient 3-D culture system design. PMID:26558344

  10. A Bacterial Continuous Culture System Based on a Microfluidic Droplet Open Reactor.

    PubMed

    Ito, Manami; Sugiura, Haruka; Ayukawa, Shotaro; Kiga, Daisuke; Takinoue, Masahiro

    2016-01-01

    Recently, micrometer-sized bacterial culture systems have attracted attention as useful tools for synthetic biology studies. Here, we present the development of a bacterial continuous culture system based on a microdroplet open reactor consisting of two types of water-in-oil microdroplets with diameters of several hundred micrometers. A continuous culture was realized the through supply of nutrient substrates and the removal of waste and excess bacterial cells based on repeated fusion and fission of droplets. The growth dynamics was controlled by the interval of fusion. We constructed a microfluidic system and quantitatively assessed the dynamics of the bacterial growth using a mathematical model. This system will facilitate the study of synthetic biology and metabolic engineering in the future. PMID:26753707

  11. An integrated microfluidic cell culture system for high-throughput perfusion three-dimensional cell culture-based assays: effect of cell culture model on the results of chemosensitivity assays.

    PubMed

    Huang, Song-Bin; Wang, Shih-Siou; Hsieh, Chia-Hsun; Lin, Yung Chang; Lai, Chao-Sung; Wu, Min-Hsien

    2013-03-21

    Although microfluidic cell culture systems are versatile tools for cellular assays, their use has yet to set in motion an evolutionary shift away from conventional cell culture methods. This situation is mainly due to technical hurdles: the operational barriers to the end-users, the lack of compatible detection schemes capable of reading out the results of a microfluidic-based cellular assay, and the lack of fundamental data to bridge the gap between microfluidic and conventional cell culture models. To address these issues, we propose a high-throughput, perfusion, three-dimensional (3-D) microfluidic cell culture system encompassing 30 microbioreactors. This integrated system not only aims to provide a user-friendly cell culture tool for biologists to perform assays but also to enable them to obtain precise data. Its technical features include (i) integration of a heater chip based on transparent indium tin oxide glass, providing stable thermal conditions for cell culturing; (ii) a microscale 3-D culture sample loading scheme that is both efficient and precise; (iii) a non-mechanical pneumatically driven multiplex medium perfusion mechanism; and (iv) a microplate reader-compatible waste medium collector array for the subsequent high throughput bioassays. In this study, we found that the 3-D culture sample loading method provided uniform sample loading [coefficient of variation (CV): 3.2%]. In addition, the multiplex medium perfusion mechanism led to reasonably uniform (CV: 3.6-6.9%) medium pumping rates in the 30 microchannels. Moreover, we used the proposed system to perform a successful cell culture-based chemosensitivity assay. To determine the effects of cell culture models on the cellular proliferation, and the results of chemosensitivity assays, we compared our data with that obtained using three conventional cell culture models. We found that the nature of the cell culture format could lead to different evaluation outcomes. Consequently, when establishing a

  12. Culture system and long-term storage of culture media in the in vitro production of bovine embryos.

    PubMed

    Varga, Santiago; Diez, Carmen; Fernández, Lina; Alvarez, Jenny; Katchicualula, Adelino; Hidalgo, Carlos Olegario; Tamargo, Carolina; Carbajo, Maite

    2011-03-01

    The optimum culture system for in vitro matured and fertilised oocytes still remains to be clarified. Culture media (CM) for mammalian embryos are routinely prepared fresh for use and preserved under refrigeration during one or two weeks. The purposes of this work were (1) to compare the efficiency of a synthetic oviduct fluid (SOF) with two different bovine serum albumin (BSA) concentrations (3 and 8 g/L) for the in vitro production of bovine blastocysts, (2) to test the effect of timing on adding fetal calf serum (FCS) to the SOF, and (3) to evaluate the effects on bovine embryo development of freezing and lyophilisation as procedures for preserving the SOF. Supplementation of SOF with 3 g/L BSA increased Day-7 blastocyst expansion rates (18.3 ± 1.6 vs. 14.4 ± 0.7; P < 0.05), although no differences in hatching rates were found. Addition of FCS to SOFaa (SOF with amino acids) medium supplemented with sodium citrate (SOFaaci) at 48 and at 72 h post-insemination (PI) allowed obtaining higher Day-6 embryo development rates than when FCS was added at 18 or 96 h PI (Day-6 morulae + blastocyst rate: 30.0 ± 1.1, 40.8 ± 1.1, 43.9 ± 2.3 and 39.3 ± 0.5 for FCS addition at 18, 48, 72 and 96 h, respectively). Hatching rates were significantly improved when serum was added at 72 h PI. Finally, both refrigeration and lyophilisation appeared as useful cryopreservation procedures for SOFaaci, although a significant loss of its ability to support embryo development, compared to the control fresh culture medium, was observed. PMID:21354948

  13. Efficient expansion of mesenchymal stromal cells in a disposable fixed bed culture system.

    PubMed

    Mizukami, Amanda; Orellana, Maristela D; Caruso, Sâmia R; de Lima Prata, Karen; Covas, Dimas T; Swiech, Kamilla

    2013-01-01

    The need for efficient and reliable technologies for clinical-scale expansion of mesenchymal stromal cells (MSC) has led to the use of disposable bioreactors and culture systems. Here, we evaluate the expansion of cord blood-derived MSC in a disposable fixed bed culture system. Starting from an initial cell density of 6.0 × 10(7) cells, after 7 days of culture, it was possible to produce of 4.2(±0.8) × 10(8) cells, which represents a fold increase of 7.0 (±1.4). After enzymatic retrieval from Fibra-Cell disks, the cells were able to maintain their potential for differentiation into adipocytes and osteocytes and were positive for many markers common to MSC (CD73, CD90, and CD105). The results obtained in this study demonstrate that MSC can be efficiently expanded in the culture system. This novel approach presents several advantages over the current expansion systems, based on culture flasks or microcarrier-based spinner flasks and represents a key element for MSC cellular therapy according to GMP compliant clinical-scale production system. PMID:23420706

  14. Culture, Indigenous Knowledge Systems and Sustainable Development: A Critical View of Education in an African Context

    ERIC Educational Resources Information Center

    Breidlid, Anders

    2009-01-01

    The article's focus is the relationship between culture, indigenous knowledge systems (IKS), sustainable development and education in Africa. It analyzes the concept of sustainability with particular reference to education and indigenous knowledge systems. In particular the article analyzes the documents from the World Summit in Johannesburg in…

  15. Organizational Culture and the Design of Computer-Mediated Communication Systems: Issues for Organizational Communication Research.

    ERIC Educational Resources Information Center

    Hacker, Kenneth L.; And Others

    The study of computer mediated communication (CMC) systems in organizations is necessary for a complete examination and explanation of organizational culture and communication. Research has shown that the effects of CMC systems have been both positive and negative. Positively, they have helped to augment oral communication. Negatively, they have…

  16. Revisiting Bourdieu: Alternative Educational Systems in the Light of the Theory of Social and Cultural Reproduction

    ERIC Educational Resources Information Center

    Azaola, Marta Cristina

    2012-01-01

    The paper reflects upon the principles and practice of an alternative educational system operating in rural Mexico in the light of Bourdieu's theory of cultural and social reproduction. Bourdieu's theory seeks to explain processes of reproduction of power relations within schools and society; whereas alternative educational systems seek to expand…

  17. Summary of the Culture, History and Educational System of Viet Nam.

    ERIC Educational Resources Information Center

    Nguyen, Chinh B., Comp.

    This booklet gives information on the culture, history and educational system of Vietnam as it relates to Vietnamese students in the American educational system. It is intended for use by teachers who have Vietnamese students in their classes. It consists of the following chapters: (1) Vietnamese Geography and History; (2) Vietnamese Family…

  18. The Transformation of Ergonomic Affordances into Cultural Affordances: The Case of the Alnuset System

    ERIC Educational Resources Information Center

    Chiappini, Giampaolo

    2012-01-01

    Is it possible to study the ergonomic affordances offered by a system designed for educational aims and their transformation into cultural affordances? To this purpose, what references can we adopt? This work describes the theoretical framework used to realise this study referring to AlNuSet, a system realised within the EC ReMath project to…

  19. Cometabolism of Monochloramine by Distribution System Relevant Mixed Culture Nitrifiers

    EPA Science Inventory

    Monochloramine (NH2Cl) is increasingly used as a residual disinfectant. A major problem related to NH2Cl is nitrification in distribution systems, leading to rapid NH2Cl residual loss. Ammonia-oxidizing bacteria (AOB), which oxidize ammonia (NH3) to nitrite, can cometabolize chem...

  20. The cultural articulation of patriarchy: legal systems, Islam and women.

    PubMed

    Shaheed, F

    1986-01-01

    Patriarchy in Pakistan results in inequalities to women. Issues emphasized include Islamic customary laws, the movement to Islamize penal and social behavioral codes, the mislabeling of Islamic beliefs as "westernization", and the rising women's movement's attempt to oppose present trends. Many practices thought to reflect Muslim culture are really the infliction of Islamic religious principles on pre-existing behavioral codes in Pakistan; thus, such practices are not actually Islamic teachings and are used to control social behavior. It is necessary to separate Islamic institution and actual Pakistanian practices in order to identify Islamic ideology's role in sustaining and vindicating patriachary. 3 roots of shaping jurisprudence are customary law, religious law, and British civil and criminal law. Further investigation of customary and religious laws currently employed indicates acceptance of Muslim practices promoting superiority of men and rejection of Islamic teachings promoting women's rights. Such Islamic teachings include a marriage settlement requiring men to give money to their wives, acknowledgement of marriages as an agreement between consenting adults, and a woman's right to divorce. Customs contradicting Islamic teachings and leading to inequalities for women include denial of a woman's access to economic resources, the annulment of the marriage settlement, and the relative ease of Muslim men to divorce their wives. Some communities practice purdah in which women are secluded from men and excluded in economic and political decisions. Such social restrictions minimize women's involvement in political decision making and in the judiciary. Exercising their right to vote and participating in trade unions, women would influence decision making. Resistance to current practices has been trade unions, women could influence decision making. resistance to current practices has been primarily from upper and middle class women; but to be effective all classes

  1. Effect of culture conditions on morphological changes of Helicobacter pylori.

    PubMed

    Tominaga, K; Hamasaki, N; Watanabe, T; Uchida, T; Fujiwara, Y; Takaishi, O; Higuchi, K; Arakawa, T; Ishii, E; Kobayashi, K; Yano, I; Kuroki, T

    1999-01-01

    The morphological conversion of Helicobacter pylori from the spiral form to the coccoid form may be the expression of a transitory adaptation to an unsuitable environment. The mechanism(s) of this conversion are not clear. In this study, we examined whether the morphological conversion of H. pylori is affected by various culture conditions, such as oxygen concentration, pH, temperature, or the presence of beta-cyclodextrin. H. pylori (NTCC11916) was cultured on Brucella agar, followed by culture in Brucella broth containing 1% agar under several conditions. Morphological conversion of individual H. pylori on the agar plate was investigated with time after incubation under phase contrast microscopy. When H. pylori was inoculated in Brucella broth containing beta-cyclodextrin, the spiral form of the organism was observed even after 6 days of incubation under standard culture conditions: 37 degrees C, pH 7, and microaerobic atmosphere (5% O2/10% CO2/85% N2) (control). The morphological conversion of H. pylori was completed on day 3 in an aerobic atmosphere (20% O2 supply) and on day 2 in an undermicroaerobic atmosphere (<0.1% O2). Its complete morphological conversion was observed at pH 8 on day 5 and at pH 4 on day 6. All of the H. pylori (100%) incubated at 20 degrees or 42 degrees C had converted from the bacillary to the coccoid form on day 4. Conditioned medium without beta-cyclodextrin caused complete conversion on day 5. These results suggest that oxygen concentration, pH, temperature, and beta-cyclodextrin may be related to the H. pylori morphological conversion from the bacillary to the coccoid form. PMID:10616762

  2. Use of hydrogen peroxide treatment and crystal violet agar plates for selective recovery of bacteriophages from natural environments

    SciTech Connect

    Asghari, A.; Farrah, S.R.; Bitton, G. )

    1992-04-01

    Hydrogen peroxide inactivated bacteriophages and bacteria at different rates. A concentration of 0.1% hydrogen peroxide reduced the numbers of several bacteria by an average of 94% but caused an average of 25% inactivation in the numbers of bacteriophages tested. Treating natural samples with hydrogen peroxide selectively reduced the indigenous bacterial flora and permitted better visualization of plaques of lawns of Escherichia coli C-3000. In some cases indigenous gram-positive bacteria were relatively resistant to hydrogen peroxide, but their growth could be limited by incorporation of crystal violet into the bottom agar used for plaque assays. The use of hydrogen peroxide treatment and crystal violet-containing plates permitted recovery of more phages from natural samples than did other procedures, such as chloroform pretreatment or the use of selective plating agar such as EC medium.

  3. Use of benzimidazole agar plates to assess fall armyworm (Lepidoptera: Noctuidae) feeding on excised maize and sorghum leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is an economically significant pest of sorghum and maize. To screen sorghum and maize germplasm for resistance to fall armyworm feeding, field, greenhouse, or lab bioassays are often utilized individually or in combinatio...

  4. Clinical comparison of the isolator and BacT/Alert aerobic blood culture systems.

    PubMed Central

    Hellinger, W C; Cawley, J J; Alvarez, S; Hogan, S F; Harmsen, W S; Ilstrup, D M; Cockerill, F R

    1995-01-01

    The performance characteristics of the Isolator (Wampole Laboratories, Cranbury, N.J.) and the BacT/Alert (Organon Teknika Corporation, Durham, N.C.) aerobic blood culture systems were compared for 6,009 blood culture sets obtained from patients with suspected bloodstream infections. The BacT/Alert aerobic bottle [BTA(O2)] was continuously agitated while it was incubated in 5% CO2 at 36 degrees C; culture plates prepared from the Isolator tube [I(O2)] were incubated in 5% CO2 at 37 degrees C. From 394 blood cultures, 416 clinically significant isolates of bacteria and yeasts were recovered. The overall yields for BTA(O2) and I(O2) were not significantly different (319 versus 336; P = 0.20). I(O2) recovered significantly more staphylococcus (P < 0.05) and yeast isolates (P < 0.01). BTA(O2) recovered significantly more aerobic and facultatively anaerobic gram-negative bacilli (P < 0.05). In blood culture sets which produced growth of the same organisms in both the BTA(O2) and I(O2) systems, the BTA(O2) system detected growth sooner, but more rapid identification was possible with the I(O2) system by virtue of earlier isolation of colonies on solid media. PMID:7665647

  5. Garlic exerts allelopathic effects on pepper physiology in a hydroponic co-culture system.

    PubMed

    Ding, Haiyan; Cheng, Zhihui; Liu, Menglong; Hayat, Sikandar; Feng, Han

    2016-01-01

    A hydroponic co-culture system was adopted to determine the allelopathic potential of garlic on the growth of pepper plants. Different numbers of garlic plants (0, 2, 4, 8 and 12) were hydroponically co-cultured with two pepper plants to investigate allelopathic effects on the growth attributes and antioxidative defense system of the test pepper plants. The responses of the pepper plants depended on the number of garlic plants included in the co-culture system, indicating an association of pepper growth with the garlic root exudate concentration. When grown at a pepper/garlic ratio of 1:1 or 1:2, the pepper plant height, chlorophyll content, and peroxidase (POD), catalase (CAT) and phenylalanine ammonia-lyase (PAL) activities were significantly increased after 30 days of co-culture; in contrast, reduction in methane dicarboxylic aldehyde (MDA) content was observed. However, when the pepper/garlic ratio was 1:4 or higher, these morphological indices and protective enzyme activities were significantly inhibited, whereas MDA levels in the pepper leaves were significantly increased due to severe membrane lipid peroxidation. The results indicate that although low concentrations of garlic root exudates appear to induce protective enzyme systems and promote pepper growth, high concentrations have deleterious effects. These findings suggest that further investigations should optimize the co-culture pepper/garlic ratio to reduce continuous cropping obstacles in pepper production. PMID:27095440

  6. Garlic exerts allelopathic effects on pepper physiology in a hydroponic co-culture system

    PubMed Central

    Ding, Haiyan; Liu, Menglong; Hayat, Sikandar; Feng, Han

    2016-01-01

    ABSTRACT A hydroponic co-culture system was adopted to determine the allelopathic potential of garlic on the growth of pepper plants. Different numbers of garlic plants (0, 2, 4, 8 and 12) were hydroponically co-cultured with two pepper plants to investigate allelopathic effects on the growth attributes and antioxidative defense system of the test pepper plants. The responses of the pepper plants depended on the number of garlic plants included in the co-culture system, indicating an association of pepper growth with the garlic root exudate concentration. When grown at a pepper/garlic ratio of 1:1 or 1:2, the pepper plant height, chlorophyll content, and peroxidase (POD), catalase (CAT) and phenylalanine ammonia-lyase (PAL) activities were significantly increased after 30 days of co-culture; in contrast, reduction in methane dicarboxylic aldehyde (MDA) content was observed. However, when the pepper/garlic ratio was 1:4 or higher, these morphological indices and protective enzyme activities were significantly inhibited, whereas MDA levels in the pepper leaves were significantly increased due to severe membrane lipid peroxidation. The results indicate that although low concentrations of garlic root exudates appear to induce protective enzyme systems and promote pepper growth, high concentrations have deleterious effects. These findings suggest that further investigations should optimize the co-culture pepper/garlic ratio to reduce continuous cropping obstacles in pepper production. PMID:27095440

  7. A modular culture system for the generation of multiple specialized tissues.

    PubMed

    Minuth, Will W; Denk, Lucia; Glashauser, Anne

    2010-04-01

    Numerous factors influence cell functions and tissue development in culture. A modular culture system has been developed to allow the control of many of these important environmental parameters. Optimal adhesion of cells is obtained by selecting an individual biomaterial. Selected specimens are mounted in a tissue carrier in order to protect it against damage during handling and after seeding cells, the carriers can be used in a series of compatible perfusion culture containers. This technique allows the simple bathing of growing tissue under continuous medium transport and the exposure of epithelia to a gradient with different fluids at the luminal and basal sides. A further container is made of transparent material to observe microscopically the developing tissue. In addition, a special model features a flexible silicone lid to apply force to mimic the mechanical load required for developing connective and muscular tissue. Perfusion culture of stem/progenitor cells at the interface of an artificial interstitium made by a polyester fleece results in the spatial development of tubules. During long term culture over weeks the growing tissue is continuously exposed to fresh nutrition and respiratory gas. The medium is transported in a constant flow or in pulses, preventing unstirred layers of fluid. A variety of applications of this modular system, described in this paper, demonstrates that the biological profile of cells and tissues can be strongly improved when perfusion culture with a permanent provision of fresh medium is applied. PMID:20096452

  8. An integrated system for synchronous culture of animal cells under controlled conditions.

    PubMed

    Mendoza-Pérez, Elena; Hernández, Vanessa; Palomares, Laura A; Serrato, José A

    2016-01-01

    The cell cycle has fundamental effects on cell cultures and their products. Tools to synchronize cultured cells allow the study of cellular physiology and metabolism at particular cell cycle phases. However, cells are most often arrested by methods that alter their homeostasis and are then cultivated in poorly controlled environments. Cell behavior could then be affected by the synchronization method and culture conditions used, and not just by the particular cell cycle phase under study. Moreover, only a few viable cells are recovered. Here, we designed an integrated system where a large number of cells from a controlled bioreactor culture is separated by centrifugal elutriation at high viabilities. In contrast to current elutriation methods, cells are injected directly from a bioreactor into an injection loop, allowing the introduction of a large number of cells into the separation chamber without stressful centrifugation. A low pulsation peristaltic pump increases the stability of the elutriation chamber. Using this approach, a large number of healthy cells at each cell cycle phase were obtained, allowing their direct inoculation into fully instrumented bioreactors. Hybridoma cells synchronized and cultured in this system behaved as expected for a synchronous culture. PMID:27625207

  9. The Porcine Aortic Tissue Culture System in vitro for Stem Cell Research

    PubMed Central

    Kim, Dong-Eun; Oh, Keun-Hee; Yang, Ji-Hye; Kwon, Sun-Keun; Cho, Tae-Jun; Lee, Seul-Bi; Nam, Hyun; Lee, Dong-Sup; Lee, Jung-Ryul; Lee, Gene

    2011-01-01

    Background and Objectives: Due to the shortage of human donors for transplantation, the use of animal organs for xenotransplantation has come into great interest. Xeno-derived vessels and cardiac valves would be possible alternatives for the patient suffering from cardiovascular diseases. Therefore, we established in vitro culture system of a porcine vessel that could be helpful for the research of xenograft and stem cell research. Methods and Results: We primarily isolated porcine thoracic aorta, cultured square-shaped pieces up to 17 days and analyzed its morphology and characters. The endothelial cells were primarily isolated from cultured porcine aortic pieces and their morphology, function and character were analyzed in order to confirm them as endothelial cells at day 3, 4, 8, 10 and 17. Even at day 17, the morphology exhibited the intact endothelial layer as well as specifically expressed CD31 and von Willebrand factor. The morphology of primarily isolated cells from cultured tissues was identical as an endothelial cell. By flow cytometry analysis, more than 80% of the isolated cells expressed CD31 and up to 80% took up acetyl low density lipoprotein (ac-LDL) until day 10 of tissue culture period even though it decreased to about 50% at day 17 that means they not only showed typical endothelial cell characters but also functioned properly. Conclusions: We successfully established and optimized a porcine vascular tissue in vitro culture system that could be a valuable model for in vitro study of xenotransplantation and stem cell research. PMID:24298344

  10. A three-dimensional culture system for the growth of hematopoietic cells.

    PubMed

    Naughton, B A; Jacob, L; Naughton, G K

    1990-01-01

    A physiological three-dimensional culture system was developed for the growth of human bone marrow. Bone marrow stromal cells were established on a nylon filtration screen template, suspended in liquid medium and grown to 70% confluence, and inoculated with hematopoietic cells. An intricate microenvironment is established to support hematopoiesis, which proceeds in a three-dimensional orientation. Analysis of the adherent zone of these cultures with flow cytometry and progenitor cell assays reveals multilineage hematologic expression and active proliferation of immature cells for the 12 week experimental period. Similar results were obtained with rat bone marrow cultures using this methodology. The suspended nylon mesh system is novel in that it supports the growth of several hematologic lineages concurrently. This system may lend itself to the growth of purged or untreated bone marrow for transplantation. PMID:2308994

  11. Proteomics analysis of nasopharyngeal carcinoma cell secretome using a hollow fiber culture system and mass spectrometry.

    PubMed

    Wu, Hsin-Yi; Chang, Ying-Hwa; Chang, Yu-Chen; Liao, Pao-Chi

    2009-01-01

    Secreted proteins, referred to as the secretome, are known to regulate a variety of biological functions and are involved in a multitude of pathological processes. However, some secreted proteins from cell cultures are difficult to detect because of their intrinsic low abundance. They are frequently masked by proteins shed from lysed cells and the substantial amounts of serum proteins used in culture medium. We have proposed an analytical platform for sensitive detection of secreted proteins by utilizing a hollow fiber culture (HFC) system coupled with proteomic approaches. The HFC system enables culture of high-density cells in a small volume where secreted proteins can be accumulated. In addition, cell lysis rates can be greatly reduced, which alleviates the contamination from lysed cells. In this study, nasopharyngeal carcinoma (NPC) cells were utilized to evaluate the efficiency of this system in the collection and analysis of the cell secretome. Cells were adapted to serum-free medium and inoculated into the HFC system. The cell lysis rate in the culture system was estimated to be 0.001-0.022%, as determined by probing four intracellular proteins in the conditioned medium (CM), while a cell lysis rate of 0.32-1.84% was observed in dish cultures. Proteins in the CM were analyzed using SDS-PAGE and liquid chromatography tandem mass spectrometry (LC-MS/MS). A total of 134 proteins were identified in 62 gel bands, of which 61% possess a signal peptide and/or a transmembrane domain. In addition, 37% of the identified secretome were classified as extracellular or membrane proteins, whereas 98% of the lysate proteins were identified as intracellular proteins. We suggest that the HFC system may be used to collect secreted proteins efficiently and facilitate comprehensive characterization of cell secretome. PMID:19012429

  12. Assessment of Long-Term Effects of Nanoparticles in a Microcarrier Cell Culture System

    PubMed Central

    Mrakovcic, Maria; Absenger, Markus; Riedl, Regina; Smole, Claudia; Roblegg, Eva; Fröhlich, Leopold F.; Fröhlich, Eleonore

    2013-01-01

    Nano-sized materials could find multiple applications in medical diagnosis and therapy. One main concern is that engineered nanoparticles, similar to combustion-derived nanoparticles, may cause adverse effects on human health by accumulation of entire particles or their degradation products. Chronic cytotoxicity must therefore be evaluated. In order to perform chronic cytotoxicity testing of plain polystyrene nanoparticles on the endothelial cell line EAhy 926, we established a microcarrier cell culture system for anchorage-dependent cells (BioLevitatorTM). Cells were cultured for four weeks and exposed to doses, which were not cytotoxic upon 24 hours of exposure. For comparison, these particles were also studied in regularly sub-cultured cells, a method that has traditionally been used to assess chronic cellular effects. Culturing on basal membrane coated microcarriers produced very high cell densities. Fluorescent particles were mainly localized in the lysosomes of the exposed cells. After four weeks of exposure, the number of cells exposed to 20 nm polystyrene particles decreased by 60% as compared to untreated controls. When tested in sub-cultured cells, the same particles decreased cell numbers to 80% of the untreated controls. Dose-dependent decreases in cell numbers were also noted after exposure of microcarrier cultured cells to 50 nm short multi-walled carbon nanotubes. Our findings support that necrosis, but not apoptosis, contributed to cell death of the exposed cells in the microcarrier culture system. In conclusion, the established microcarrier model appears to be more sensitive for the identification of cellular effects upon prolonged and repeated exposure to nanoparticles than traditional sub-culturing. PMID:23457616

  13. Assessment of long-term effects of nanoparticles in a microcarrier cell culture system.

    PubMed

    Mrakovcic, Maria; Absenger, Markus; Riedl, Regina; Smole, Claudia; Roblegg, Eva; Fröhlich, Leopold F; Fröhlich, Eleonore

    2013-01-01

    Nano-sized materials could find multiple applications in medical diagnosis and therapy. One main concern is that engineered nanoparticles, similar to combustion-derived nanoparticles, may cause adverse effects on human health by accumulation of entire particles or their degradation products. Chronic cytotoxicity must therefore be evaluated. In order to perform chronic cytotoxicity testing of plain polystyrene nanoparticles on the endothelial cell line EAhy 926, we established a microcarrier cell culture system for anchorage-dependent cells (BioLevitator(TM)). Cells were cultured for four weeks and exposed to doses, which were not cytotoxic upon 24 hours of exposure. For comparison, these particles were also studied in regularly sub-cultured cells, a method that has traditionally been used to assess chronic cellular effects. Culturing on basal membrane coated microcarriers produced very high cell densities. Fluorescent particles were mainly localized in the lysosomes of the exposed cells. After four weeks of exposure, the number of cells exposed to 20 nm polystyrene particles decreased by 60% as compared to untreated controls. When tested in sub-cultured cells, the same particles decreased cell numbers to 80% of the untreated controls. Dose-dependent decreases in cell numbers were also noted after exposure of microcarrier cultured cells to 50 nm short multi-walled carbon nanotubes. Our findings support that necrosis, but not apoptosis, contributed to cell death of the exposed cells in the microcarrier culture system. In conclusion, the established microcarrier model appears to be more sensitive for the identification of cellular effects upon prolonged and repeated exposure to nanoparticles than traditional sub-culturing. PMID:23457616

  14. The Impact of National Cultural Differences on Nurses' Acceptance of Hospital Information Systems.

    PubMed

    Lin, Hsien-Cheng

    2015-06-01

    This study aims to explore the influence of national cultural differences on nurses' perceptions of their acceptance of hospital information systems. This study uses the perspective of Technology Acceptance Model; national cultural differences in terms of masculinity/femininity, individualism/collectivism, power distance, and uncertainty avoidance are incorporated into the Technology Acceptance Model as moderators, whereas time orientation is a control variable on hospital information system acceptance. A quantitative research design was used in this study; 261 participants, US and Taiwan RNs, all had hospital information system experience. Data were collected from November 2013 to February 2014 and analyzed using a t test to compare the coefficients for each moderator. The results show that individualism/collectivism, power distance, and uncertainty avoidance all exhibit significant difference on hospital information system acceptance; however, both masculinity/femininity and time orientation factors did not show significance. This study verifies that national cultural differences have significant influence on nurses' behavioral intention to use hospital information systems. Therefore, hospital information system providers should emphasize the way in which to integrate different technological functions to meet the needs of nurses from various cultural backgrounds. PMID:25899441

  15. Modelling and pathway identification involving the transport mechanism of a complex metabolic system in batch culture

    NASA Astrophysics Data System (ADS)

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

    2014-06-01

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

  16. Evaluation of mono or mixed cultures of lactic acid bacteria in type II sourdough system.

    PubMed

    Ekinci, Raci; Şimşek, Ömer; Küçükçuban, Ayca; Nas, Sebahattin

    2016-04-01

    The aim of this study was to evaluate the use of mono and mixed lactic acid bacteria (LAB) cultures to determine suitable LAB combinations for a type II sourdough system. In this context, previously isolated sourdough LAB strains with antimicrobial activity, which included Lactobacillus plantarum PFC22, Lactobacillus brevis PFC31, Pediococcus acidilactici PFC38, and Lactobacillus sanfranciscensis PFC80, were used as mono or mixed culture combinations in a fermentation system to produce type II sourdough, and subsequently in bread dough production. Compared to the monoculture fermentation of dough, the use of mixed cultures shortened the adaptation period by half. In addition, the use of mixed cultures ensured higher microbial viability, and enhanced the fruity flavor during bread dough production. It was determined that the combination of L. plantarum PFC22 + P. acidilactici PFC38 + L. sanfranciscensis PFC80 is a promising culture mixture that can be used in the production of type II sourdough systems, and that may also contribute to an increase in metabolic activity during bread production process. PMID:25807196

  17. In Vivo Culture System Using the INVOcell Device Shows Similar Pregnancy and Implantation Rates to Those Obtained from In Vivo Culture System in ICSI Procedures

    PubMed Central

    García-Ferreyra, Javier; Hilario, Roly; Luna, Daniel; Villegas, Lucy; Romero, Rocío; Zavala, Patricia; Dueñas-Chacón, Julio

    2015-01-01

    CAPSULE Clinical outcomes using INVOcell device with ICSI. OBJECTIVE Intravaginal culture of oocytes (INVO) procedure is an intravaginal culture system that utilizes the INVOcell device in which the fertilization and embryo culture occur. In this procedure, the vaginal cavity serves as an incubator for oocyte fertilization and early embryonic development. The objective of this study was to evaluate the clinical outcomes of this intravaginal culture system in intracytoplasmic sperm injection (ICSI). METHODS A total of 24 cycles INVO-ICSI (study group) and 74 cycles of ICSI (control group) were included in the study. The cleaved oocytes at day 3/total injected oocytes, embryo quality, pregnancy rate (PR), implantation rate (IR), and miscarriage rate (MR) were compared between both groups. RESULTS At day 3, there was no difference in the cleaved oocyte rate (78.7 and 76.1%) and embryo quality (77 and 86.8%) for the study and control groups, respectively. In the study group, more embryos were significantly transferred compared to the control group (2.63 ± 0.58 versus 1.93 ± 0.25; P < 0.05). PRs, IRs, and MRs were similar for the study group compared with the control group (PR: 54.2% versus 58.1%; IR: 31.7% versus 33.6%; MR: 7.7% versus 20.9%). CONCLUSIONS Good PR and IR can be obtained using the INVOcell device, and the INVO-ICSI procedure can be considered as an alternative option to infertile patients. PMID:26085790

  18. Revised culture-based system for identification of Malassezia species.

    PubMed

    Kaneko, Takamasa; Makimura, Koichi; Abe, Michiko; Shiota, Ryoko; Nakamura, Yuka; Kano, Rui; Hasegawa, Atsuhiko; Sugita, Takashi; Shibuya, Shuichi; Watanabe, Shinichi; Yamaguchi, Hideyo; Abe, Shigeru; Okamura, Noboru

    2007-11-01

    Forty-six strains of Malassezia spp. with atypical biochemical features were isolated from 366 fresh clinical isolates from human subjects and dogs. Isolates obtained in this study included 2 (4.7%) lipid-dependent M. pachydermatis isolates; 1 (2.4%) precipitate-producing and 6 (14.6%) non-polyethoxylated castor oil (Cremophor EL)-assimilating M. furfur isolates; and 37 (34.3%) M. slooffiae isolates that were esculin hydrolyzing, 17 (15.7%) that were non-tolerant of growth at 40 degrees C, and 2 (1.9%) that assimilated polyethoxylated castor oil. Although their colony morphologies and sizes were characteristic on CHROMagar Malassezia medium (CHROM), all strains of M. furfur developed large pale pink and wrinkled colonies, and all strains of M. slooffiae developed small (<1 mm) pale pink colonies on CHROM. These atypical strains were distinguishable by the appearance of their colonies grown on CHROM. Three clinically important Malassezia species, M. globosa, M. restricta, and M. furfur, were correctly identified by their biochemical characteristics and colony morphologies. The results presented here indicate that our proposed identification system will be useful as a routine tool for the identification of clinically important Malassezia species in clinical laboratories. PMID:17881545

  19. An Improved Culture Method for Selective Isolation of Campylobacter jejuni from Wastewater.

    PubMed

    Kim, Jinyong; Oh, Euna; Banting, Graham S; Braithwaite, Shannon; Chui, Linda; Ashbolt, Nicholas J; Neumann, Norman F; Jeon, Byeonghwa

    2016-01-01

    Campylobacter jejuni is one of the leading foodborne pathogens worldwide. C. jejuni is isolated from a wide range of foods, domestic animals, wildlife, and environmental sources. The currently available culture-based isolation methods are not highly effective for wastewater samples due to the low number of C. jejuni in the midst of competing bacteria. To detect and isolate C. jejuni from wastewater samples, in this study, we evaluated a few different enrichment conditions using five different antibiotics (i.e., cefoperazone, vancomycin, trimethoprim, polymyxin B, and rifampicin), to which C. jejuni is intrinsically resistant. The selectivity of each enrichment condition was measured with C t value using quantitative real-time PCR, and multiplex PCR to determine Campylobacter species. In addition, the efficacy of Campylobacter isolation on different culture media after selective enrichment was examined by growing on Bolton and Preston agar plates. The addition of polymyxin B, rifampicin, or both to the Bolton selective supplements enhanced the selective isolation of C. jejuni. The results of 16S rDNA sequencing also revealed that Enterococcus spp. and Pseudomonas aeruginosa are major competing bacteria in the enrichment conditions. Although it is known to be difficult to isolate Campylobacter from samples with heavy contamination, this study well exhibited that the manipulation of antibiotic selective pressure improves the isolation efficiency of fastidious Campylobacter from wastewater. PMID:27617011

  20. An Improved Culture Method for Selective Isolation of Campylobacter jejuni from Wastewater

    PubMed Central

    Kim, Jinyong; Oh, Euna; Banting, Graham S.; Braithwaite, Shannon; Chui, Linda; Ashbolt, Nicholas J.; Neumann, Norman F.; Jeon, Byeonghwa

    2016-01-01

    Campylobacter jejuni is one of the leading foodborne pathogens worldwide. C. jejuni is isolated from a wide range of foods, domestic animals, wildlife, and environmental sources. The currently available culture-based isolation methods are not highly effective for wastewater samples due to the low number of C. jejuni in the midst of competing bacteria. To detect and isolate C. jejuni from wastewater samples, in this study, we evaluated a few different enrichment conditions using five different antibiotics (i.e., cefoperazone, vancomycin, trimethoprim, polymyxin B, and rifampicin), to which C. jejuni is intrinsically resistant. The selectivity of each enrichment condition was measured with Ct value using quantitative real-time PCR, and multiplex PCR to determine Campylobacter species. In addition, the efficacy of Campylobacter isolation on different culture media after selective enrichment was examined by growing on Bolton and Preston agar plates. The addition of polymyxin B, rifampicin, or both to the Bolton selective supplements enhanced the selective isolation of C. jejuni. The results of 16S rDNA sequencing also revealed that Enterococcus spp. and Pseudomonas aeruginosa are major competing bacteria in the enrichment conditions. Although it is known to be difficult to isolate Campylobacter from samples with heavy contamination, this study well exhibited that the manipulation of antibiotic selective pressure improves the isolation efficiency of fastidious Campylobacter from wastewater. PMID:27617011

  1. PCR-Based Multiple Species Cell Counting for In Vitro Mixed Culture

    PubMed Central

    Huang, Ruijie; Zhang, Junjie; Yang, X. Frank; Gregory, Richard L.

    2015-01-01

    Changes of bacterial profiles in microbial communities are strongly associated with human health. There is an increasing need for multiple species research in vitro. To avoid high cost or measurement of a limited number of species, PCR-based multiple species cell counting (PCR-MSCC) has been conceived. Species-specific sequence is defined as a unique sequence of one species in a multiple species mixed culture. This sequence is identified by comparing a random 1000 bp genomic sequence of one species with the whole genome sequences of the other species in the same artificial mixed culture. If absent in the other genomes, it is the species-specific sequence. Species-specific primers were designed based on the species-specific sequences. In the present study, ten different oral bacterial species were mixed and grown in Brain Heart Infusion Yeast Extract with 1% sucrose for 24 hours. Biofilm was harvested and processed for DNA extraction and q-PCR amplification with the species-specific primers. By comparing the q-PCR data of each species in the unknown culture with reference cultures, in which the cell number of each species was determined by colony forming units on agar plate, the cell number of that strain in the unknown mixed culture was calculated. This technique is reliable to count microorganism numbers that are less than 100,000 fold different from other species within the same culture. Theoretically, it can be used in detecting a species in a mixed culture of over 200 species. Currently PCR-MSCC is one of the most economic methods for quantifying single species cell numbers, especially for the low abundant species, in a multiple artificial mixed culture in vitro. PMID:25970462

  2. The Clinical Urine Culture: Enhanced Techniques Improve Detection of Clinically Relevant Microorganisms.

    PubMed

    Price, Travis K; Dune, Tanaka; Hilt, Evann E; Thomas-White, Krystal J; Kliethermes, Stephanie; Brincat, Cynthia; Brubaker, Linda; Wolfe, Alan J; Mueller, Elizabeth R; Schreckenberger, Paul C

    2016-05-01

    Enhanced quantitative urine culture (EQUC) detects live microorganisms in the vast majority of urine specimens reported as "no growth" by the standard urine culture protocol. Here, we evaluated an expanded set of EQUC conditions (expanded-spectrum EQUC) to identify an optimal version that provides a more complete description of uropathogens in women experiencing urinary tract infection (UTI)-like symptoms. One hundred fifty adult urogynecology patient-participants were characterized using a self-completed validated UTI symptom assessment (UTISA) questionnaire and asked "Do you feel you have a UTI?" Women responding negatively were recruited into the no-UTI cohort, while women responding affirmatively were recruited into the UTI cohort; the latter cohort was reassessed with the UTISA questionnaire 3 to 7 days later. Baseline catheterized urine samples were plated using both standard urine culture and expanded-spectrum EQUC protocols: standard urine culture inoculated at 1 μl onto 2 agars incubated aerobically; expanded-spectrum EQUC inoculated at three different volumes of urine onto 7 combinations of agars and environments. Compared to expanded-spectrum EQUC, standard urine culture missed 67% of uropathogens overall and 50% in participants with severe urinary symptoms. Thirty-six percent of participants with missed uropathogens reported no symptom resolution after treatment by standard urine culture results. Optimal detection of uropathogens could be achieved using the following: 100 μl of urine plated onto blood (blood agar plate [BAP]), colistin-nalidixic acid (CNA), and MacConkey agars in 5% CO2 for 48 h. This streamlined EQUC protocol achieved 84% uropathogen detection relative to 33% detection by standard urine culture. The streamlined EQUC protocol improves detection of uropathogens that are likely relevant for symptomatic women, giving clinicians the opportunity to receive additional information not currently reported using standard urine culture

  3. Using a 3D Culture System to Differentiate Visceral Adipocytes In Vitro.

    PubMed

    Emont, Margo P; Yu, Hui; Jun, Heejin; Hong, Xiaowei; Maganti, Nenita; Stegemann, Jan P; Wu, Jun

    2015-12-01

    It has long been recognized that body fat distribution and regional adiposity play a major role in the control of metabolic homeostasis. However, the ability to study and compare the cell autonomous regulation and response of adipocytes from different fat depots has been hampered by the difficulty of inducing preadipocytes isolated from the visceral depot to differentiate into mature adipocytes in culture. Here, we present an easily created 3-dimensional (3D) culture system that can be used to differentiate preadipocytes from the visceral depot as robustly as those from the sc depot. The cells differentiated in these 3D collagen gels are mature adipocytes that retain depot-specific characteristics, as determined by imaging, gene expression, and functional assays. This 3D culture system therefore allows for study of the development and function of adipocytes from both depots in vitro and may ultimately lead to a greater understanding of site-specific functional differences of adipose tissues to metabolic dysregulation. PMID:26425808

  4. 3D culture of ovarian follicles: a system towards their engineering?

    PubMed

    Zuccotti, Maurizio; Merico, Valeria; Rebuzzini, Paola; Belli, Martina; Vigone, Giulia; Mulas, Francesca; Fassina, Lorenzo; Wruck, Wasco; Adjaye, James; Bellazzi, Riccardo; Garagna, Silvia

    2015-01-01

    Infertility in women is a health priority. Designing a robust culture protocol capable of attaining complete follicle growth is an exciting challenge, for its potential clinical applications, but also as a model to observe and closely study the sequence of molecular events that lie behind the intricate relationship existing between the oocyte and surrounding follicle cells. Here, we describe the procedures used to maintain the ovarian follicle 3D architecture employing a variety of in vitro systems and several types of matrices. Collagen and alginate are the matrices that led to better results, including proof-of-concept of full-term development. Pioneer in its kind, these studies underlie the drawbacks encountered and the need for a culture system that allows more quantitative analyses and predictions, projecting the culture of the ovarian follicle into the realm of tissue engineering. PMID:26505254

  5. A pneumatic pressure-driven multi-throughput microfluidic circulation culture system.

    PubMed

    Satoh, T; Narazaki, G; Sugita, R; Kobayashi, H; Sugiura, S; Kanamori, T

    2016-06-21

    Here, we report a pneumatic pressure-driven microfluidic device capable of multi-throughput medium circulation culture. The circulation culture system has the following advantages for application in drug discovery: (i) simultaneous operation of multiple circulation units, (ii) use of a small amount of circulating medium (3.5 mL), (iii) pipette-friendly liquid handling, and (iv) a detachable interface with pneumatic pressure lines via sterile air-vent filters. The microfluidic device contains three independent circulation culture units, in which human umbilical vein endothelial cells (HUVECs) were cultured under physiological shear stress induced by circulation of the medium. Circulation of the medium in the three culture units was generated by programmed sequentially applied pressure from two pressure-control lines. HUVECs cultured in the microfluidic device were aligned under a one-way circulating flow with a shear stress of 10 dyn cm(-2); they exhibited a randomly ordered alignment under no shear stress and under reciprocating flow with a shear stress of 10 dyn cm(-2). We also observed 2.8- to 4.9-fold increases in expression of the mRNAs of endothelial nitric oxide synthase and thrombomodulin under one-way circulating flow with a shear stress of 10 dyn cm(-2) compared with conditions of no shear stress or reciprocating flow. PMID:27229626

  6. Three-dimensional Huh7 cell culture system for the study of Hepatitis C virus infection

    PubMed Central

    Sainz, Bruno; TenCate, Veronica; Uprichard, Susan L

    2009-01-01

    Background In order to elucidate how Hepatitis C Virus (HCV) interacts with polarized hepatocytes in vivo and how HCV-induced alterations in cellular function contribute to HCV-associated liver disease, a more physiologically relevant hepatocyte culture model is needed. As such, NASA-engineered three-dimensional (3-D) rotating wall vessel (RWV) bioreactors were used in effort to promote differentiation of HCV-permissive Huh7 hepatoma cells. Results When cultured in the RWV, Huh7 cells became morphologically and transcriptionally distinct from more standard Huh7 two-dimensional (2-D) monolayers. Specifically, RWV-cultured Huh7 cells formed complex, multilayered 3-D aggregates in which Phase I and Phase II xenobiotic drug metabolism genes, as well as hepatocyte-specific transcripts (HNF4α, Albumin, TTR and α1AT), were upregulated compared to 2-D cultured Huh7 cells. Immunofluorescence analysis revealed that these HCV-permissive 3-D cultured Huh7 cells were more polarized than their 2D counterparts with the expression of HCV receptors, cell adhesion and tight junction markers (CD81, scavenger receptor class B member 1, claudin-1, occludin, ZO-1, β-Catenin and E-Cadherin) significantly increased and exhibiting apical, lateral and/or basolateral localization. Conclusion These findings show that when cultured in 3-D, Huh7 cells acquire a more differentiated hepatocyte-like phenotype. Importantly, we show that these 3D cultures are highly permissive for HCV infection, thus providing an opportunity to study HCV entry and the effects of HCV infection on host cell function in a more physiologically relevant cell culture system. PMID:19604376

  7. The potential use of constructed wetlands in a recirculating aquaculture system for shrimp culture.

    PubMed

    Lin, Ying-Feng; Jing, Shuh-Ren; Lee, Der-Yuan

    2003-01-01

    A pilot-scale constructed wetland unit, consisting of free water surface (FWS) and subsurface flow (SF) constructed wetlands arranged in series, was integrated into an outdoor recirculating aquaculture system (RAS) for culturing Pacific white shrimp (Litopenaeus vannamei). This study evaluated the performance of the wetland unit in treating the recirculating wastewater and examined the effect of improvement in water quality of the culture tank on the growth and survival of shrimp postlarvae. During an 80-day culture period, the wetland unit operated at a mean hydraulic loading rate of 0.3 m/day and effectively reduced the influent concentrations of 5-day biochemical oxygen demand (BOD5, 24%), suspended solids (SS, 71%), chlorophyll a (chl-a, 88%), total ammonium (TAN, 57%), nitrite nitrogen (NO2-N, 90%) and nitrate nitrogen (NO3-N, 68%). Phosphate (PO4-P) reduction was the least efficient (5.4%). The concentrations of SS, Chl-a, turbidity and NO3-N in the culture tank water in RAS were significantly (Psystem (CAS) that simulated static pond culture without wetland treatment. However, no significant difference (Psystems. At the end of the study, the harvest results showed that shrimp weight and survival rate in the RAS (3.8 +/-1.8 g/shrimp and 90%) significantly (Pculture environment, consequently increasing the shrimp growth and survival without water exchange, in a recirculating system. PMID:12663210

  8. Three-dimensional hydrogel cell culture systems for modeling neural tissue

    NASA Astrophysics Data System (ADS)

    Frampton, John

    Two-dimensional (2-D) neural cell culture systems have served as physiological models for understanding the cellular and molecular events that underlie responses to physical and chemical stimuli, control sensory and motor function, and lead to the development of neurological diseases. However, the development of three-dimensional (3-D) cell culture systems will be essential for the advancement of experimental research in a variety of fields including tissue engineering, chemical transport and delivery, cell growth, and cell-cell communication. In 3-D cell culture, cells are provided with an environment similar to tissue, in which they are surrounded on all sides by other cells, structural molecules and adhesion ligands. Cells grown in 3-D culture systems display morphologies and functions more similar to those observed in vivo, and can be cultured in such a way as to recapitulate the structural organization and biological properties of tissue. This thesis describes a hydrogel-based culture system, capable of supporting the growth and function of several neural cell types in 3-D. Alginate hydrogels were characterized in terms of their biomechanical and biochemical properties and were functionalized by covalent attachment of whole proteins and peptide epitopes. Methods were developed for rapid cross-linking of alginate hydrogels, thus permitting the incorporation of cells into 3-D scaffolds without adversely affecting cell viability or function. A variety of neural cell types were tested including astrocytes, microglia, and neurons. Cells remained viable and functional for longer than two weeks in culture and displayed process outgrowth in 3-D. Cell constructs were created that varied in cell density, type and organization, providing experimental flexibility for studying cell interactions and behavior. In one set of experiments, 3-D glial-endothelial cell co-cultures were used to model blood-brain barrier (BBB) structure and function. This co-culture system was

  9. Culture characteristics of the atmospheric and room temperature plasma-mutated Spirulina platensis mutants in CO2 aeration culture system for biomass production.

    PubMed

    Tan, Yinyee; Fang, Mingyue; Jin, Lihua; Zhang, Chong; Li, He-Ping; Xing, Xin-Hui

    2015-10-01

    For biomass production of Spirulina platensis as feedstock of fermentation, the culture characteristics of three typical mutants of 3-A10, 3-B2 and 4-B3 generated by atmospheric and room temperature plasmas (ARTP) mutagenesis were systematically studied by using CO2 aeration culture system and compared with the wild strain. The specific growth rate of wild strain in the pure air aeration culture system exhibited a 76.2% increase compared with static culture, while the specific growth rates of the 3-A10, 3-B2 and 4-B3 in pure air aeration culture system were increased by 114.4%, 95.9% and 88.2% compared with their static cultures. Compared with static culture, the carbohydrate contents of wild strain, 3-A10, 3-B2 and 4-B3 in pure air aeration culture system dropped plainly by 51.0%, 79.3%, 85.5% and 26.1%. Increase of CO2 concentration enhanced carbohydrate content and productivity. Based on the carbohydrate productivity, the optimal inlet of CO2 concentration in aeration culture was determined to be 12% (v/v). Under this condition, 3-B2 exhibited the highest carbohydrate content (30.7%), CO2 fixation rate (0.120gCO2·g(-1)·d(-1)) and higher growth rate (0.093 g L(-1)·d(-1)), while 3-A10 showed the highest growth rate (0.118 g L(-1)·d(-1)) and higher CO2 fixation rate (0.117gCO2·g(-1)·d(-1)) but low carbohydrate content (24.5%), and 4-B3 showed the highest chlorophyll (Chl) content (3.82 mg·g(-1)). The most outstanding mutant by static culture in terms of growth rate and carbohydrate productivity (3-B2), was also demonstrated by CO2 aeration culture system. This study revealed that the ARTP mutagenesis could generate the S. platensis mutants suitable for CO2 aeration culture aiming at biomass production. PMID:25795571

  10. Culturing thick brain slices: An interstitial 3D microperfusion system for enhanced viability

    PubMed Central

    Rambani, Komal; Vukasinovic, Jelena; Glezer, Ari; Potter, Steve M.

    2009-01-01

    Brain slice preparations are well-established models for a wide spectrum of in vitro investigations in the neuroscience discipline. However, these investigations are limited to acute preparations or thin organotypic culture preparations due to the lack of a successful method that allows culturing of thick organotypic brain slices. Thick brain slice cultures suffer necrosis due to ischemia deep in the tissue resulting from a destroyed circulatory system and subsequent diffusion-limited supply of nutrients and oxygen. Although thin organotypic brain slice cultures can be successfully cultured using a well established roller tube method (a monolayer organotypic culture) (Gahwiler B H, 1981) or a membrane insert method (up to 1–4 cell layers, <150μm)(Stoppini L et al., 1991), these methods fail to support thick tissue preparations. A few perfusion methods (using submerged or interface/microfluidic chambers) have been reported to enhance the longevity (up to few hours) of acute slice preparations (up to 600μm thick) (Hass H L et al., 1979; Nicoll R A and Alger B E, 1981; Passeraub P A et al., 2003). Here, we report a unique interstitial microfluidic perfusion technique to culture thick (700μm) organotypic brain slices. The design of the custom-made micro-perfusion chamber facilitates laminar, interstitial perfusion of oxygenated nutrient medium throughout the tissue thickness with concomitant removal of depleted medium and catabolites. We examined the utility of this perfusion method to enhance the viability of the thick organotypic brain slice cultures after 2 days and 5 days in vitro (DIV). We investigated the range of amenable flow rates that enhance the viability of 700μm thick organotypic brain slices compared to the unperfused control cultures. Our perfusion method allows up to 84.6% viability (P<0.01) and up to 700μm thickness, even after 5 DIV. Our results also confirm that these cultures are functionally active and have their in vivo cytoarchitecture

  11. A primary culture system of mouse thick ascending limb cells with preserved function and uromodulin processing.

    PubMed

    Glaudemans, Bob; Terryn, Sara; Gölz, Nadine; Brunati, Martina; Cattaneo, Angela; Bachi, Angela; Al-Qusairi, Lama; Ziegler, Urs; Staub, Olivier; Rampoldi, Luca; Devuyst, Olivier

    2014-02-01

    The epithelial cells lining the thick ascending limb (TAL) of the loop of Henle perform essential transport processes and secrete uromodulin, the most abundant protein in normal urine. The lack of differentiated cell culture systems has hampered studies of TAL functions. Here, we report a method to generate differentiated primary cultures of TAL cells, developed from microdissected tubules obtained in mouse kidneys. The TAL tubules cultured on permeable filters formed polarized confluent monolayers in ∼12 days. The TAL cells remain differentiated and express functional markers such as uromodulin, NKCC2, and ROMK at the apical membrane. Electrophysiological measurements on primary TAL monolayers showed a lumen-positive transepithelial potential (+9.4 ± 0.8 mV/cm(2)) and transepithelial resistance similar to that recorded in vivo. The transepithelial potential is abolished by apical bumetanide and in primary cultures obtained from ROMK knockout mice. The processing, maturation and apical secretion of uromodulin by primary TAL cells is identical to that observed in vivo. The primary TAL cells respond appropriately to hypoxia, hypertonicity, and stimulation by desmopressin, and they can be transfected. The establishment of this primary culture system will allow the investigation of TAL cells obtained from genetically modified mouse models, providing a critical tool for understanding the role of that segment in health and disease. PMID:23887378

  12. A Defined, Controlled Culture System for Primary Bovine Chromaffin Progenitors Reveals Novel Biomarkers and Modulators

    PubMed Central

    Masjkur, Jimmy; Levenfus, Ian; Lange, Sven; Arps-Forker, Carina; Poser, Steve; Qin, Nan; Vukicevic, Vladimir; Chavakis, Triantafyllos; Eisenhofer, Graeme; Bornstein, Stefan R.; Ehrhart-Bornstein, Monika

    2014-01-01

    We present a method to efficiently culture primary chromaffin progenitors from the adult bovine adrenal medulla in a defined, serum-free monolayer system. Tissue is dissociated and plated for expansion under support by the mitogen basic fibroblast growth factor (bFGF). The cultures, although not homogenous, contain a subpopulation of cells expressing the neural stem cell marker Hes3 that also propagate. In addition, Hes3 is also expressed in the adult adrenal medulla from where the tissue is taken. Differentiation is induced by bFGF withdrawal and switching to Neurobasal medium containing B27. Following differentiation, Hes3 expression is lost, and cells acquire morphologies and biomarker expression patterns of chromaffin cells and dopaminergic neurons. We tested the effect of different treatments that we previously showed regulate Hes3 expression and cell number in cultures of fetal and adult rodent neural stem cells. Treatment of the cultures with a combination of Delta4, Angiopoietin2, and a Janus kinase inhibitor increases cell number during the expansion phase without significantly affecting catecholamine content levels. Treatment with cholera toxin does not significantly affect cell number but reduces the ratio of epinephrine to norepinephrine content and increases the dopamine content relative to total catecholamines. These data suggest that this defined culture system can be used for target identification in drug discovery programs and that the transcription factor Hes3 may serve as a new biomarker of putative adrenomedullary chromaffin progenitor cells. PMID:24855275

  13. Co-culture systems and technologies: taking synthetic biology to the next level

    PubMed Central

    Goers, Lisa; Freemont, Paul; Polizzi, Karen M.

    2014-01-01

    Co-culture techniques find myriad applications in biology for studying natural or synthetic interactions between cell populations. Such techniques are of great importance in synthetic biology, as multi-species cell consortia and other natural or synthetic ecology systems are widely seen to hold enormous potential for foundational research as well as novel industrial, medical and environmental applications with many proof-of-principle studies in recent years. What is needed for co-cultures to fulfil their potential? Cell–cell interactions in co-cultures are strongly influenced by the extracellular environment, which is determined by the experimental set-up, which therefore needs to be given careful consideration. An overview of existing experimental and theoretical co-culture set-ups in synthetic biology and adjacent fields is given here, and challenges and opportunities involved in such experiments are discussed. Greater focus on foundational technology developments for co-cultures is needed for many synthetic biology systems to realize their potential in both applications and answering biological questions. PMID:24829281

  14. Co-culture systems and technologies: taking synthetic biology to the next level.

    PubMed

    Goers, Lisa; Freemont, Paul; Polizzi, Karen M

    2014-07-01

    Co-culture techniques find myriad applications in biology for studying natural or synthetic interactions between cell populations. Such techniques are of great importance in synthetic biology, as multi-species cell consortia and other natural or synthetic ecology systems are widely seen to hold enormous potential for foundational research as well as novel industrial, medical and environmental applications with many proof-of-principle studies in recent years. What is needed for co-cultures to fulfil their potential? Cell-cell interactions in co-cultures are strongly influenced by the extracellular environment, which is determined by the experimental set-up, which therefore needs to be given careful consideration. An overview of existing experimental and theoretical co-culture set-ups in synthetic biology and adjacent fields is given here, and challenges and opportunities involved in such experiments are discussed. Greater focus on foundational technology developments for co-cultures is needed for many synthetic biology systems to realize their potential in both applications and answering biological questions. PMID:24829281

  15. From the World-Systems Perspective to Institutional World History: Culture and Economy in Global Theory.

    ERIC Educational Resources Information Center

    Benton, Lauren

    1996-01-01

    Summarizes critical attacks on Immanuel Wallerstein's World Systems approach to history and offers new critical evaluations. Wallerstein argued that an emerging capitalist world economy dominated politics and history from the 16th century to the present. Defines two new approaches, institutional analysis and post colonial cultural theory, that…

  16. Hybrid System for the Inventory of the Cultural Heritage Using Voice Interface for Knowledge acquisition

    NASA Astrophysics Data System (ADS)

    Château, Stefan Du; Boulanger, Danielle; Mercier-Laurent, Eunika

    This document presents our work on a definition and experimentation of a voice interface for cultural heritage inventory. This hybrid system includes signal processing, natural language techniques and knowledge modeling for future retrieval. We discuss the first results and present some challenges for our future work.

  17. Cultural Capital in East Asian Educational Systems: The Case of Japan

    ERIC Educational Resources Information Center

    Yamamoto, Yoko; Brinton, Mary C.

    2010-01-01

    Cultural capital has been an important but often elusive concept in the study of educational processes and social class reproduction. The authors suggest that this is partly because a country's educational system and ways of evaluating students at different educational transitions set the context for the mechanisms through which embodied and…

  18. Development of Quality Assurance System in Culture and Nation Character Education in Primary Education in Indonesia

    ERIC Educational Resources Information Center

    Susilana, Rudi; Asra

    2013-01-01

    The purpose of national education is to develop skills and build dignified national character and civilization in educating nation life (Act No. 20, 2003). The paper describes a system of quality assurance in culture and character education in primary education. This study employs the six sigma model which consists of the formula DMAIC (Define,…

  19. The Learning Transfer System Inventory (LTSI) in Ukraine: The Cross-Cultural Validation of the Instrument

    ERIC Educational Resources Information Center

    Yamkovenko, Bogdan V.; Holton, Elwood, III; Bates, R. A.

    2007-01-01

    Purpose: The purpose of this research is to expand cross-cultural research and validate the Learning Transfer System Inventory in Ukraine. The researchers seek to translate the LTSI into Ukrainian and investigate the internal structure of this translated version of the questionnaire. Design/methodology/approach: The LTSI is translated into…

  20. PLANT CULTURAL SYSTEM FOR MONITORING EVAPOTRANSPIRATION AND PHYSIOLOGICAL RESPONSES UNDER FIELD CONDITIONS

    EPA Science Inventory

    A plant culture system incorporating the water-table root-screen method for controlling plant water status was adapted for use in open-top field exposure chambers for studying the effects of drought stress on physiological responses. The daily transpiration rates of the plants we...

  1. Raising Cultural Self-Efficacy among Faculty and Staff of a Private Native Hawaiian School System

    ERIC Educational Resources Information Center

    Fong, Randie Kamuela

    2012-01-01

    The Hawaiian cultural revitalization movement in Hawai`i is an important driver for many Hawaiian organizations as well as educational institutions that serve Native Hawaiians. One such organization is Kamehameha Schools, a private school system founded and endowed by Princess Bernice Pauahi Bishop in 1887 to educate Native Hawaiian children. From…

  2. Improving Achievement for Linguistically and Culturally Diverse Learners through an Inquiry-Based Earth Systems Curriculum

    ERIC Educational Resources Information Center

    Lambert, Julie; Ariza, Eileen N. Whelan

    2008-01-01

    This report describes an inquiry-based Earth systems curriculum and strategies for teaching diverse students, which were embedded in the curriculum. The curriculum was implemented with 5th-grade students with varied linguistic, cultural, and socioeconomic backgrounds in five schools in a large, southeastern U.S., urban school district. At the end…

  3. Social-Cultural-Historical Contradictions in an L2 Listening Lesson: A Joint Activity System Analysis

    ERIC Educational Resources Information Center

    Cross, Jeremy

    2011-01-01

    Informed and inspired by neo-Vygotskian theory, this article outlines a study exploiting a contemporary conceptualization of Wells's (2002) joint activity system model as an exploratory framework for examining and depicting the social-cultural-historical contradictions in second-language (L2) learners' joint activity. The participants were a pair…

  4. National Contexts Influencing Principals' Time Use and Allocation: Economic Development, Societal Culture, and Educational System

    ERIC Educational Resources Information Center

    Lee, Moosung; Hallinger, Philip

    2012-01-01

    This study examines the impact of macro-context factors on the behavior of school principals. More specifically, the article illuminates how a nation's level of economic development, societal culture, and educational system influence the amount of time principals devote to their job role and shape their allocation of time to instructional…

  5. FURTHER DEVELOPMENT OF RODENT WHOLE EMBRYO CULTURE: SOLVENT TOXICITY AND WATER INSOLUBLE COMPOUND DELIVERY SYSTEM

    EPA Science Inventory

    In order to study the in vitro embryotoxicity and dysmorphogenesis of water insoluble compounds, solvents or chemical delivery systems of low toxicity and teratogenicity to the developing embryo must be found. Therefore, day 10.5 rat embryos were cultured for 2 days in whole rat ...

  6. Enforced Cultural Change in Academe. A Practical Case Study: Implementing Management Systems in Higher Education.

    ERIC Educational Resources Information Center

    Spencer-Matthews, Sarah

    2001-01-01

    An action research project sought to implement a quality management system in an academic department in an Australian higher education institution. The case study revealed that technical change was attained, but adoption of a quality culture achieved only token acceptance. (EV)

  7. Cultural Influences and Corporate Decision Making: The Humanities/Information Systems Partnership.

    ERIC Educational Resources Information Center

    Skovira, Robert J.

    Robert Morris College received a national Endowment for the Humanities grant to create a faculty study project to internationalize the humanities curriculum. The Humanities can play a role in building the cultural contexts and contributing to a deeper understanding of information-based corporate decision making in Information Systems courses,…

  8. Organizing the Baby Boomer Construct: An Exploration of Marketing, Social Systems, and Culture

    ERIC Educational Resources Information Center

    Lipschultz, Jeremy H.; Hilt, Michael L.; Reilly, Hugh J.

    2007-01-01

    Baby boomer trends are applied in the development of a conceptual framework that offers a social systems and cultural model for future studies. While there has been considerable recent attention paid to baby boomers, the studies lack a coherent theoretical base that would allow for more advanced and continuing research. Aging baby boomers heading…

  9. Spearmint plantlet culture system as a means to study secondary metabolism in vitro

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The production of phytochemicals in vitro has been obtained from a variety of tissue types and organs. A plantlet culture system offers a means to study whole plant growth and development in a miniature scale and their corresponding phytochemical production. Plantlets resemble their in vivo counte...

  10. Effect of cultural system and essential oil treatment on antioxidant capacity in raspberries

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effects of cultural system and essential oil treatment on antioxidant enzyme activities, antioxidant capacities and flavonoid contents in raspberries were evaluated. Raspberries were hand-harvested from organic and conventional farms in Maryland, USA, and were treated with essential oils includi...

  11. Improving Hospital Quality and Patient Safety an Examination of Organizational Culture and Information Systems

    ERIC Educational Resources Information Center

    Gardner, John Wallace

    2012-01-01

    This dissertation examines the effects of safety culture, including operational climate and practices, as well as the adoption and use of information systems for delivering high quality healthcare and improved patient experience. Chapter 2 studies the influence of both general and outcome-specific hospital climate and quality practices on process…

  12. Comparison of the Expression of Hepatic Genes by Human Wharton’s Jelly Mesenchymal Stem Cells Cultured in 2D and 3D Collagen Culture Systems

    PubMed Central

    Khodabandeh, Zahra; Vojdani, Zahra; Talaei-Khozani, Tahereh; Jaberipour, Mansoureh; Hosseini, Ahmad; Bahmanpour, Soghra

    2016-01-01

    Background: Human Wharton’s jelly mesenchymal stem cells (HWJMSCs) express liver-specific markers such as albumin, alpha-fetoprotein, cytokeratin-19, cytokeratin-18, and glucose-6-phosphatase. Therefore, they can be considered as a good source for cell replacement therapy for liver diseases. This study aimed to evaluate the effects of various culture systems on the hepatocyte-specific gene expression pattern of naïve HWJMSCs. Methods: HWJMSCs were characterized as MSCs by detecting the surface CD markers and capability to differentiate toward osteoblast and adipocyte. HWJMSCs were cultured in 2D collagen films and 3D collagen scaffolds for 21 days and were compared to control cultures. Real time RT-PCR was used to evaluate the expression of liver-specific genes. Results: The HWJMSCs which were grown on non-coated culture plates expressed cytokeratin-18 and -19, alpha-fetoprotein, albumin, glucose-6-phosphatase, and claudin. The expression of the hepatic nuclear factor 4 (HNF4) was very low. The cells showed a significant increase in caludin expression when they cultured in 3D collagen scaffolds compared to the conventional monolayer culture and 2D collagen scaffold. Conclusion: Various culture systems did not influence on hepatocyte specific marker expression by HWJMSCs, except for claudin. The expression of claudin showed that 3D collagen scaffold provided the extracellular matrix for induction of the cells to interconnect with each other. PMID:26722142

  13. Enrichment and Molecular Characterization of a Bacterial Culture That Degrades Methoxy-Methyl Urea Herbicides and Their Aniline Derivatives

    PubMed Central

    El-Fantroussi, Said

    2000-01-01

    Soil treated with linuron for more than 10 years showed high biodegradation activity towards methoxy-methyl urea herbicides. Untreated control soil samples taken from the same location did not express any linuron degradation activity, even after 40 days of incubation. Hence, the occurrence in the field of a microbiota having the capacity to degrade a specific herbicide was related to the long-term treatment of the soil. The enrichment culture isolated from treated soil showed specific degradation activity towards methoxy-methyl urea herbicides, such as linuron and metobromuron, while dimethyl urea herbicides, such as diuron, chlorotoluron, and isoproturon, were not transformed. The putative metabolic intermediates of linuron and metobromuron, the aniline derivatives 3,4-dichloroaniline and 4-bromoaniline, were also degraded. The temperature of incubation drastically affected degradation of the aniline derivatives. Whereas linuron was transformed at 28 and 37°C, 3,4-dichloroaniline was transformed only at 28°C. Monitoring the enrichment process by reverse transcription-PCR and denaturing gradient gel electrophoresis (DGGE) showed that a mixture of bacterial species under adequate physiological conditions was required to completely transform linuron. This research indicates that for biodegradation of linuron, several years of adaptation have led to selection of a bacterial consortium capable of completely transforming linuron. Moreover, several of the putative species appear to be difficult to culture since they were detectable by DGGE but were not culturable on agar plates. PMID:11097876

  14. An ex-ovo chicken embryo culture system suitable for imaging and microsurgery applications.

    PubMed

    Yalcin, Huseyin C; Shekhar, Akshay; Rane, Ajinkya A; Butcher, Jonathan T

    2010-01-01

    Understanding the relationships between genetic and microenvironmental factors that drive normal and malformed embryonic development is fundamental for discovering new therapeutic strategies. Advancements in imaging technology have enabled quantitative investigation of the organization and maturing of the body plan, but later stage embryonic morphogenesis is less clear. Chicken embryos are an attractive vertebrate animal model system for this application because of its ease of culture and surgical manipulation. Early embryos can be cultured for a short time on filter paper rings, which enables complete optical access for cell patterning and fate studies. Studying advanced developmental processes such as cardiac morphogenesis are traditionally performed through a window of the eggshell, but this technique limits optical access due to window size. We previously developed a simple method to culture whole embryos ex-ovo on hexagonal weigh boats for up to 10 days, which enabled high resolution imaging via ultrasonography. These cultures were difficult to transport, limiting the types of imaging tools available for live experiments. We here present an improved shell-less culture system with a cost-effective, portable environmental chamber. Eggs were cracked onto a hammock created by a polyurethane membrane (cling wrap) affixed circumferentially to a plastic cup partially filled with sterile water. The dimensions of the circumference and depth of the hammock were both critical to maintain surface tension, while the mechanics of the hammock and water beneath helped dampen vibrations induced by transportation. A small footprint circulating water bath was also developed to enable continuous temperature control during experimentation. We demonstrate the ability to culture embryos in this way for at least 14 days without morphogenic defect or delay and employ this system in several microsurgical and imaging applications. PMID:21048670

  15. Recurrent amoebic gill infestation in rainbow trout cultured in a semiclosed water recirculation system

    USGS Publications Warehouse

    Noble, A.C.; Herman, R.L.; Noga, E.J.; Bullock, G.L.

    1997-01-01

    Five lots of commercially purchased juvenile rainbow trout Oncorhynchus mykiss (17-44 g) stocked in a continuous-production water recirculation system became infested with gilt amoebae. The amoebae were introduced into the recirculation system, as evidenced by their presence on gills of fish held in quarantine tanks. Based on their morphology, as seen in histological sections and by electron microscopy, the amoebae appeared to be more closely related to the family Cochliopodiidae than to other taxa of free living amoebae. Attempts to culture the amoebae in different media, at different temperatures of incubation, and in fish cell culture were not successful. Initial treatment of the recirculation system with formalin at 167 parts per million (ppm) for 1 h eliminated amoebae from the gills. Subsequent treatments of the entire system with formalin at 50-167 ppm reduced the intensity of further infestations.

  16. COMBINED USE OF A WATER-INSOLUBLE CHEMICAL DELIVERY SYSTEM AND A METABOLIC ACTIVATION SYSTEM IN WHOLE EMBRYO CULTURE

    EPA Science Inventory

    An integrated water insoluble chemical delivery/metabolic activation/rat embryo culture system is described. In initial studies corn oil was used as the solvent and diallate as the substrate. Increasing concentrations of diallate dissolved in corn oil caused embryonic growth reta...

  17. Phosphorylation of the insulin receptor in cultured hepatoma cells and a solubilized system

    SciTech Connect

    Kasuga, M.; White, M.F.; Kahn, C.R.

    1985-01-01

    Methods are described which have been used successfully to study insulin receptor autophosphorylation in cultured cells (hepatoma cell line Fao) and detergent solubilized receptor systems. Intact cultured cells were labelled with /sup 32/PO/sub 4//sup 3 -/. Details are given for the solubilization and purification of the insulin receptor and insulin dose-response curves for phosphorylation of the solubilized insulin receptor. Trypsin digestion of a phosphorylated subunit suggests that at least peptides containing sites of /sup 32/P incorporation exist in the receptor molecule.

  18. Enhanced Geothermal System Development of the AmeriCulture Leasehold in the Animas Valley

    SciTech Connect

    Duchane, David V; Seawright, Gary L; Sewright, Damon E; Brown, Don; Witcher, James c.; Nichols, Kenneth E.

    2001-03-02

    Working under the grant with AmeriCulture, Inc., and its team of geothermal experts, assembled a plan to apply enhanced geothermal systems (EGS) techniques to increase both the temperature and flow rate of the geothermal waters on its leasehold. AmeriCulture operates a commercial aquaculture facility that will benefit from the larger quantities of thermal energy and low cost electric power that EGS technology can provide. The project brought together a team of specialists that, as a group, provided the full range of expertise required to successfully develop and implement the project.

  19. The culture of Chlorella vulgaris with human urine in multibiological life support system experiments

    NASA Astrophysics Data System (ADS)

    Li, Ming; Liu, Hong; Tong, Ling; Fu, Yuming; He, Wenting; Hu, Enzhu; Hu, Dawei

    The Integrative Experimental System (IES) was established as a tool to evaluate the rela-tionship of the subsystems in Bioregenerative Life Support System, and Multibiological Life Support System Experiments (MLSSE) have been conducted in the IES. The IES consists of a higher plant chamber, an animal chamber and a plate photo bioreactor (PPB) which cultivated lettuce (Lactuca sativa L.), silkworm (Bombyx Mori L.) and microalgae (Chlorella vulgaris), respectively. In MLSSE, four volunteers took turns breathing the system air through a tube connected with the animal chamber periodically. According to the CO2 concentration in the IES, the automotive control system of the PPB changed the light intensity regulating the photosynthesis of Chlorella vulgaris to make CO2 /O2 in the system maintain at stable levels. Chlorella vulgaris grew with human urine by carrying certain amount of alga liquid out of the bioreactor every day with synthetic urine replenished into the system, and O2 was regenerated, at the same time human urine was purified. Results showed that this IES worked stably and Chlorella vulgaris grew well; The culture of Chlorella vulgaris could be used to keep the balance of CO2 and O2 , and the change of light intensity could control the gas composition in the IES; Microalgae culture could be used in emergency in the system, the culture of Chlorella vulgaris could recover to original state in 5 days; 15.6 ml of condensation water was obtained every day by the culture of Chlorella vulgaris; The removal efficiencies of N, P in human urine could reach to 98.2% and 99.5%.

  20. Incubator embedded cell culture imaging system (EmSight) based on Fourier ptychographic microscopy.

    PubMed

    Kim, Jinho; Henley, Beverley M; Kim, Charlene H; Lester, Henry A; Yang, Changhuei

    2016-08-01

    Multi-day tracking of cells in culture systems can provide valuable information in bioscience experiments. We report the development of a cell culture imaging system, named EmSight, which incorporates multiple compact Fourier ptychographic microscopes with a standard multiwell imaging plate. The system is housed in an incubator and presently incorporates six microscopes. By using the same low magnification objective lenses as the objective and the tube lens, the EmSight is configured as a 1:1 imaging system that, providing large field-of-view (FOV) imaging onto a low-cost CMOS imaging sensor. The EmSight improves the image resolution by capturing a series of images of the sample at varying illumination angles; the instrument reconstructs a higher-resolution image by using the iterative Fourier ptychographic algorithm. In addition to providing high-resolution brightfield and phase imaging, the EmSight is also capable of fluorescence imaging at the native resolution of the objectives. We characterized the system using a phase Siemens star target, and show four-fold improved coherent resolution (synthetic NA of 0.42) and a depth of field of 0.2 mm. To conduct live, long-term dopaminergic neuron imaging, we cultured ventral midbrain from mice driving eGFP from the tyrosine hydroxylase promoter. The EmSight system tracks movements of dopaminergic neurons over a 21 day period. PMID:27570701

  1. Incubator embedded cell culture imaging system (EmSight) based on Fourier ptychographic microscopy

    PubMed Central

    Kim, Jinho; Henley, Beverley M.; Kim, Charlene H.; Lester, Henry A.; Yang, Changhuei

    2016-01-01

    Multi-day tracking of cells in culture systems can provide valuable information in bioscience experiments. We report the development of a cell culture imaging system, named EmSight, which incorporates multiple compact Fourier ptychographic microscopes with a standard multiwell imaging plate. The system is housed in an incubator and presently incorporates six microscopes. By using the same low magnification objective lenses as the objective and the tube lens, the EmSight is configured as a 1:1 imaging system that, providing large field-of-view (FOV) imaging onto a low-cost CMOS imaging sensor. The EmSight improves the image resolution by capturing a series of images of the sample at varying illumination angles; the instrument reconstructs a higher-resolution image by using the iterative Fourier ptychographic algorithm. In addition to providing high-resolution brightfield and phase imaging, the EmSight is also capable of fluorescence imaging at the native resolution of the objectives. We characterized the system using a phase Siemens star target, and show four-fold improved coherent resolution (synthetic NA of 0.42) and a depth of field of 0.2 mm. To conduct live, long-term dopaminergic neuron imaging, we cultured ventral midbrain from mice driving eGFP from the tyrosine hydroxylase promoter. The EmSight system tracks movements of dopaminergic neurons over a 21 day period. PMID:27570701

  2. A bladder cancer microenvironment simulation system based on a microfluidic co-culture model.

    PubMed

    Liu, Peng-fei; Cao, Yan-wei; Zhang, Shu-dong; Zhao, Yang; Liu, Xiao-guang; Shi, Hao-qing; Hu, Ke-yao; Zhu, Guan-qun; Ma, Bo; Niu, Hai-tao

    2015-11-10

    A tumor microenvironment may promote tumor metastasis and progression through the dynamic interplay between neoplastic cells and stromal cells. In this work, the most representative and significant stromal cells, fibroblasts, endothelial cells, and macrophages were used as vital component elements and combined with bladder cancer cells to construct a bladder cancer microenvironment simulation system. This is the first report to explore bladder cancer microenvironments based on 4 types of cells co-cultured simultaneously. This simulation system comprises perfusion equipment, matrigel channel units, a medium channel and four indirect contact culture chambers, allowing four types of cells to simultaneously interact through soluble biological factors and metabolites. With this system, bladder cancer cells (T24) with a tendency to form a 'reticular' structure under 3 dimensional culture conditions were observed in real time. The microenvironment characteristics of paracrine interactions and cell motility were successfully simulated in this system. The phenotype change process in stromal cells was successfully reproduced in this system by testing the macrophage effector molecule Arg-1. Arg-1 was highly expressed in the simulated tumor microenvironment group. To develop "precision medicine" in bladder cancer therapy, bladder cancer cells were treated with different clinical 'neo-adjuvant' chemotherapy schemes in this system, and their sensitivity differences were fully reflected. This work provides a preliminary foundation for neo-adjuvant chemotherapy in bladder cancer, a theoretical foundation for tumor microenvironment simulation and promotes individual therapy in bladder cancer patients. PMID:26462177

  3. Use of a Closed Culture System to Improve the Safety of Lentiviral Vector Production.

    PubMed

    Wu, Tao; Bour, Gaëtan; Durand, Sarah; Lindner, Véronique; Gossé, Francine; Zona, Laetitia; Certoux, Jean-Marie; Diana, Michele; Baumert, Thomas F; Marescaux, Jacques; Mutter, Didier; Pessaux, Patrick; Robinet, Eric

    2015-12-01

    We evaluated the possibility of introducing a combination of six oncogenes into primary porcine hepatocytes (PPH) using a lentiviral vector (LV)-mediated gene transfer in order to develop a porcine hepatocellular carcinoma model based on autologous transplantation of ex vivo-transformed hepatocytes. The six oncogenes were introduced into three plasmids, hence enabling the production of LVs encoding a luciferase reporter gene and hTERT+p53(DD), cyclinD1+CDK4(R24C), and c-myc(T58A)+HRas(G21V) genes, respectively. In order to improve the protection of the laboratory personnel manipulating such LVs, we used a compact cell culture cassette (CliniCell(®) device) as a closed cell culture system. We demonstrated that the CliniCell device allows to produce LVs, through plasmid transfection of 293T cells, and, after transfer to a second cassette, to transduce PPH with a similar efficacy as conventional open cell culture systems such as flasks or Petri dishes. Additionally, it is possible to cryopreserve at -80°C the transduced cells, directly in the CliniCell device used for the transduction. In conclusion, the use of a closed culture system for the safe handling of oncogene-encoding LVs lays the foundation for the development of porcine tumor models based on the autologous transplantation of ex vivo-transformed primary cells. PMID:26467420

  4. Modelling of Microalgae Culture Systems with Applications to Control and Optimization.

    PubMed

    Bernard, Olivier; Mairet, Francis; Chachuat, Benoît

    2016-01-01

    Mathematical modeling is becoming ever more important to assess the potential, guide the design, and enable the efficient operation and control of industrial-scale microalgae culture systems (MCS). The development of overall, inherently multiphysics, models involves coupling separate submodels of (i) the intrinsic biological properties, including growth, decay, and biosynthesis as well as the effect of light and temperature on these processes, and (ii) the physical properties, such as the hydrodynamics, light attenuation, and temperature in the culture medium. When considering high-density microalgae culture, in particular, the coupling between biology and physics becomes critical. This chapter reviews existing models, with a particular focus on the Droop model, which is a precursor model, and it highlights the structure common to many microalgae growth models. It summarizes the main developments and difficulties towards multiphysics models of MCS as well as applications of these models for monitoring, control, and optimization purposes. PMID:25604163

  5. Prolonged survival of virulent Treponema pallidum (Nichols strain) in cell-free and tissue culture systems.

    PubMed

    Fieldsteel, A H; Becker, F A; Stout, J G

    1977-10-01

    Survival of Treponema pallidum was found to be prolonged in the presence of tissue culture. Of the 12 cultures studied, cottontail rabbit epithelium (Sf1Ep) supported T. pallidum for the longest time. In horizontal Leighton tubes with reduced medium and an atmosphere of 5% CO2 in N2, the 50% survival time (ST50) was 5 to 6 days for treponemes associated with monolayers of Sf1Ep cells. Comparable cell-free tubes had ST50 values of less than 4 days. In vertical Leighton tubes containing 6 ml of prereduced medium incubated aerobically, gradients of O2 tension and redox potential were established. Attachment and survival of T. pallidum were greatest at a depth of about 10 to 20 mm. Motility was between 70 and 95% in this area throughout the first 14 days of incubation. Occasionally, greater than 50% motility was observed for as long as 21 days. The redox potential and O2 tension in the optimal area of gradient cultures were reproduced by adjusting the medium depth in a shell vial culture system containing cells on a horizontal cover slip. Treponemes associated with the cell monolayer in both gradient and shell vial cultures were still virulent after 21 days in vitro. The dilution of testis extract and the concentration of T. pallidum were found to be important factors in survival of T. pallidum. PMID:332639

  6. Assessment of patient safety culture in clinical laboratories in the Spanish National Health System

    PubMed Central

    Giménez-Marín, Angeles; Rivas-Ruiz, Francisco; García-Raja, Ana M.; Venta-Obaya, Rafael; Fusté-Ventosa, Margarita; Caballé-Martín, Inmaculada; Benítez-Estevez, Alfonso; Quinteiro-García, Ana I.; Bedini, José Luis; León-Justel, Antonio; Torra-Puig, Montserrat

    2015-01-01

    Introduction There is increasing awareness of the importance of transforming organisational culture in order to raise safety standards. This paper describes the results obtained from an evaluation of patient safety culture in a sample of clinical laboratories in public hospitals in the Spanish National Health System. Material and methods A descriptive cross-sectional study was conducted among health workers employed in the clinical laboratories of 27 public hospitals in 2012. The participants were recruited by the heads of service at each of the participating centers. Stratified analyses were performed to assess the mean score, standardized to a base of 100, of the six survey factors, together with the overall patient safety score. Results 740 completed questionnaires were received (88% of the 840 issued). The highest standardized scores were obtained in Area 1 (individual, social and cultural) with a mean value of 77 (95%CI: 76-78), and the lowest ones, in Area 3 (equipment and resources), with a mean value of 58 (95%CI: 57-59). In all areas, a greater perception of patient safety was reported by the heads of service than by other staff. Conclusions We present the first multicentre study to evaluate the culture of clinical safety in public hospital laboratories in Spain. The results obtained evidence a culture in which high regard is paid to safety, probably due to the pattern of continuous quality improvement. Nevertheless, much remains to be done, as reflected by the weaknesses detected, which identify areas and strategies for improvement. PMID:26525595

  7. Spatially Selective Reagent Delivery into Cancer Cells Using a Two-Layer Microfluidic Culture System

    PubMed Central

    Liu, Yan; Butler, W. Boyd; Pappas, Dimitri

    2012-01-01

    In this work, we demonstrate a two-layer microfluidic system capable of spatially selective delivery of drugs and other reagents under low shear stress. Loading occurs by hydrodynamically focusing a reagent stream over a particular region of the cell culture. The system consisted of a cell culture chamber and fluid flow channel, which were located in different layers to reduce shear stress on cells. Cells in the center of the culture chamber were exposed to parallel streams of laminar flow, which allowed fast changes to be made to the cellular environment. The shear force was reduced to 2.7 dyn/cm2 in the two-layer device (vs. 6.0 dyn/cm2 in a one-layer device). Cells in the side of the culture chamber were exposed to the side streams of buffer; the shear force was further reduced to a greater extent since the sides of the culture chamber were separated from the main fluid path. The channel shape and flow rate of the multiple streams were optimized for spatially-controlled reagent delivery. The boundaries between streams were well controlled at a flow rate of 0.1 mL/h, which was optimized for all streams. We demonstrated multi-reagent delivery to different regions of the same culture well, as well as selective treatment of cancer cells with a built in control group in the same well. In the case of apoptosis induction using staurosporine, 10% of cells remained viable after 24 hours of exposure. Cells in the same chamber, but not exposed to staurosporine, had a viability of 90%. This chip allows dynamic observation of cellular behavior immediately after drug delivery, as well as long-term drug treatment with the benefit of large cell numbers, device simplicity, and low shear stress. PMID:22882832

  8. Clinical laboratory comparison of the 10-ml isolator blood culture system with BACTEC radiometric blood culture media.

    PubMed

    Kellogg, J A; Manzella, J P; McConville, J H

    1984-10-01

    The efficiency of the 10-ml Isolator (E. I. du Pont de Nemours & Co., Inc.) for recovery of pathogens from blood was compared with that of BACTEC 6B and 7C media (Johnston Laboratories) by using 4,195 cultures from 1,662 patients. During the first phase of the study, BACTEC bottles were inoculated with 3 ml of blood; in the second phase, bottles were inoculated with 5 ml. The objectives were to compare results with similar blood volumes used for the detection of anaerobes as well as similar overall volumes and to determine the relative sensitivity of BACTEC media inoculated with the minimum and maximum volumes suggested by the manufacturer. From 180 patients, 391 significant isolates were recovered, 354 (91%) with the Isolator and 304 (78%) with the bottles. Isolators recovered 31 (15%) and 19 (18%) more pathogens overall than did the two-bottle system inoculated with 3 and 5 ml of blood, respectively, including 30 (36%) and 10 (34%) more Enterobacteriaceae. Recovery of anaerobes was greater in the BACTEC anaerobic medium, but only when its inoculum was increased to 5 ml. No significant differences existed between the two systems in pathogen detection times or detection of polymicrobic bacteremia. The Isolator contamination rate (8.3%) was approximately 4 times that of the bottles. The number of CFU of pathogen per milliliter of blood, blood volume sampled, and number of Isolators collected were more important than antimicrobial agent pretreatment in contributing to patient bacteremia of fungemia undetected by the Isolator. The Isolator appeared to be a practical alternative for recovery of aerobic and facultatively anaerobic pathogens from the blood. PMID:6386871

  9. Design and Physical Characterization of a Synchronous Multivalve Aortic Valve Culture System

    PubMed Central

    Durst, Christopher A.; Grande-Allen, K. Jane

    2015-01-01

    For many tissues, cyclic mechanical stimulation is considered necessary to maintain the normal morphology in vitro. The aim of this study was to design and evaluate a simple bioreactor system capable of medium-term (more than 2 weeks) culture of native and engineered aortic valves. The system consists of three pistons in separate cylindrical chambers that are simultaneously driven through the culture medium by a crank and cam assembly. The faces of these pistons have unidirectional valves mounted in opposing orientations that permit flow from one side of the face to the other. A custom designed stent was employed to secure either native or engineered trileaflet valves to the pistons. Computational fluid dynamics and finite element modeling was used to assist selection of materials and components in the system. Finally, sterility testing using base culture medium was performed to verify the ability of the system to retain sterile conditions. The current design permits the cyclic opening and closing of three aortic valves, however this device can be modified to accommodate up to 12 valves simultaneously. This new bioreactor system has applications not only for development of tissue-engineered valves, but for also studying disease models in the aortic valve. PMID:19953323

  10. Response of sago pondweed, a submerged aquatic macrophyte, to herbicides in three laboratory culture systems

    USGS Publications Warehouse

    Fleming, W.J.; Ailstock, M.S.; Momot, J.J.; Norman, C.M.

    1991-01-01

    The phytotoxicity of atrazine, paraquat, glyphosate, and alachlor to sago pondweed (Potamogeton pectinatus), a submerged aquatic macrophyte, was tested under three types of laboratory culture conditions. In each case, tests were conducted in static systems, the test period was four weeks, and herbicide exposure was chronic, resulting from a single addition of herbicide to the test vessels at the beginning of the test period. The three sets of test conditions employed were(1) axenic cultures in 125-mL flasks containing a nutrient media and sucrose; (2) a microcosm system employing 18.9-L buckets containing a sand, shell, and peat substrate; and (3) an algae-free system employing O.95-L jars containing reconstituted freshwater and a nutrient agar substrate. The primary variable measured was biomass production. Plants grew well in all three test systems, with biomass of untreated plants increasing by a factor of about 5 to 6.5 during the four-week test period. Biomass production in response to herbicide exposure differed significantly among culture systems, which demonstrates the need for a standardized testing protocol for evaluating the effects of toxics on submerged aquatic plants.

  11. The Impact of Corporate Culture, the Reward System, and Perceived Moral Intensity on Marketing Students' Ethical Decision Making

    ERIC Educational Resources Information Center

    Nill, Alexander; Schibrowsky, John A.

    2005-01-01

    An experiment was conducted to study how marketing students' ethical decision making was influenced by their perceived moral intensity (PMI), corporate culture, and the reward system. The findings indicate that levels of awareness of the ethical consequences of a decision, the corporate culture, and the reward system all significantly affect…

  12. System for exposing cultured cells to intermittent hypoxia utilizing gas permeable cultureware.

    PubMed

    Polak, Jan; Studer-Rabeler, Karen; McHugh, Holly; Hussain, Mehboob A; Shimoda, Larissa A

    2015-07-01

    Tissue intermittent hypoxia (IH) occurs in obstructive sleep apnea, sickle cell anemia, physical exercise and other conditions. Poor gas solubility and slow diffusion through culture media hampers mimicking IH-induced transitions of O(2) in vitro. We aimed to develop a system enabling exposure of cultured cells to IH and to validate such exposure by real-time O(2) measurements and cellular responses. Standard 24-well culture plates and plates with bottoms made from a gas permeable film were placed in a heated cabinet. Desired cycling of O(2) levels was induced using programmable solenoids to purge mixtures of 95% N(2) + 5% CO(2) or 95% O(2) + 5% CO(2). Dissolved oxygen, gas pressure, temperature, and water evaporation were measured during cycling. IH-induced cellular effects were evaluated by hypoxia inducible factor (HIF) and NF-κB luciferase reporters in HEK296 cells and by insulin secretion in rat insulinoma cells. Oxygen cycling in the cabinet was translated into identical changes of O(2) at the well bottom in gas permeable, but not in standard cultureware. Twenty-four hours of IH exposure increased HIF (112%), NF-κB (111%) and insulin secretion (44%). Described system enables reproducible and prolonged IH exposure in cultured cells while controlling for important environmental factors. PMID:25816360

  13. A three-dimensional culture system recapitulates placental syncytiotrophoblast development and microbial resistance

    PubMed Central

    McConkey, Cameron A.; Delorme-Axford, Elizabeth; Nickerson, Cheryl A.; Kim, Kwang Sik; Sadovsky, Yoel; Boyle, Jon P.; Coyne, Carolyn B.

    2016-01-01

    In eutherians, the placenta acts as a barrier and conduit at the maternal-fetal interface. Syncytiotrophoblasts, the multinucleated cells that cover the placental villous tree surfaces of the human placenta, are directly bathed in maternal blood and are formed by the fusion of progenitor cytotrophoblasts that underlie them. Despite their crucial role in fetal protection, many of the events that govern trophoblast fusion and protection from microbial infection are unknown. We describe a three-dimensional (3D)–based culture model using human JEG-3 trophoblast cells that develop syncytiotrophoblast phenotypes when cocultured with human microvascular endothelial cells. JEG-3 cells cultured in this system exhibit enhanced fusogenic activity and morphological and secretory activities strikingly similar to those of primary human syncytiotrophoblasts. RNASeq analyses extend the observed functional similarities to the transcriptome, where we observed significant overlap between syncytiotrophoblast-specific genes and 3D JEG-3 cultures. Furthermore, JEG-3 cells cultured in 3D are resistant to infection by viruses and Toxoplasma gondii, which mimics the high resistance of syncytiotrophoblasts to microbial infections in vivo. Given that this system is genetically manipulatable, it provides a new platform to dissect the mechanisms involved in syncytiotrophoblast development and microbial resistance. PMID:26973875

  14. An optogenetic approach for assessing formation of neuronal connections in a co-culture system.

    PubMed

    Su, Colin T E; Yoon, Su-In; Marcy, Guillaume; Chin, Eunice W M; Augustine, George J; Goh, Eyleen L K

    2015-01-01

    Here we describe a protocol to generate a co-culture consisting of 2 different neuronal populations. Induced pluripotent stem cells (iPSCs) are reprogrammed from human fibroblasts using episomal vectors. Colonies of iPSCs can be observed 30 days after initiation of fibroblast reprogramming. Pluripotent colonies are manually picked and grown in neural induction medium to permit differentiation into neural progenitor cells (NPCs). iPSCs rapidly convert into neuroepithelial cells within 1 week and retain the capability to self-renew when maintained at a high culture density. Primary mouse NPCs are differentiated into astrocytes by exposure to a serum-containing medium for 7 days and form a monolayer upon which embryonic day 18 (E18) rat cortical neurons (transfected with channelrhodopsin-2 (ChR2)) are added. Human NPCs tagged with the fluorescent protein, tandem dimer Tomato (tdTomato), are then seeded onto the astrocyte/cortical neuron culture the following day and allowed to differentiate for 28 to 35 days. We demonstrate that this system forms synaptic connections between iPSC-derived neurons and cortical neurons, evident from an increase in the frequency of synaptic currents upon photostimulation of the cortical neurons. This co-culture system provides a novel platform for evaluating the ability of iPSC-derived neurons to create synaptic connections with other neuronal populations. PMID:25742527

  15. Characterization of a primary brown adipocyte culture system derived from human fetal interscapular fat

    PubMed Central

    Seiler, Sarah E; Xu, Dan; Ho, Jia-Pei; Lo, Kinyui Alice; Buehrer, Benjamin M; Ludlow, Y John W; Kovalik, Jean-Paul; Sun, Lei

    2015-01-01

    Brown fat has gained widespread attention as a potential therapeutic target to treat obesity and associated metabolic disorders. Indeed, the anti-obesity potential of multiple targets to stimulate both brown adipocyte differentiation and recruitment have been verified in rodent models. However, their therapeutic potential in humans is unknown due to the lack of a human primary brown adipocyte cell culture system. Likewise, the lack of a well-characterized human model has limited the discovery of novel targets for the activation of human brown fat. To address this current need, we aimed to identify and describe the first primary brown adipocyte cell culture system from human fetal interscapular brown adipose tissue. Pre-adipocytes isolated from non-viable human fetal interscapular tissue were expanded and cryopreserved. Cells were then thawed and plated alongside adult human subcutaneous and omental pre-adipocytes for subsequent differentiation and phenotypic characterization. Interscapular pre-adipocytes in cell culture differentiated into mature adipocytes that were morphologically indistinguishable from the adult white depots. Throughout differentiation, cultured human fetal interscapular adipocytes demonstrated increased expression of classical brown fat markers compared to subcutaneous and omental cells. Further, functional analysis revealed an elevation in fatty acid oxidation as well as maximal and uncoupled oxygen consumption in interscapular brown adipocytes compared to white control cells. These data collectively identify the brown phenotype of these cells. Thus, our primary cell culture system derived from non-viable human fetal interscapular brown adipose tissue provides a valuable tool for the study of human brown adipocyte biology and for the development of anti-obesity therapeutics. PMID:26451287

  16. An Evaluation of Matrix-Containing and Humanised Matrix-Free 3-Dimensional Cell Culture Systems for Studying Breast Cancer

    PubMed Central

    Roberts, Grace C.; Morris, Paul G.; Moss, Marcus A.; Maltby, Sarah L.; Palmer, Chelsea A.; Nash, Claire E.; Smart, Emily; Holliday, Deborah L.; Speirs, Valerie

    2016-01-01

    Background 3D cell cultures are emerging as more physiologically meaningful alternatives to monolayer cultures for many biological applications. They are attractive because they more closely mimic in vivo morphology, especially when co-cultured with stromal fibroblasts. Methodology/Principal Findings We compared the efficacy of 3 different 3D cell culture systems; collagen I, low attachment culture vessels and a modification of Fibrolife®, a specialised humanised cell culture medium devoid of animal-derived components, using breast cancer cell lines representative of the different molecular subtypes of breast cancer, cultured alone or with human mammary fibroblasts with a view to developing matrix-free humanised systems. 3D collagen I culture supported the growth of a range of breast cancer cell lines. By modifying the composition of Fibrolife® to epiFL, matrix-free cell culture was possible. During sequential transfer to epiFL breast cancer cells gradually detached from the flask, growing progressively as spheroids. Phenotype was stable and reversible with cells remaining actively proliferating and easily accessible throughout culture. They could also be revived from frozen stocks. To achieve co-culture with fibroblasts in epiFL required use of low attachment culture vessels instead of standard plastic as fibroblasts remained adherent in epiFL. Here, cancer cell spheroids were allowed to form before adding fibroblasts. Immunohistochemical examination showed fibroblasts scattered throughout the epithelial spheroid, not dissimilar to the relationship of tumour stroma in human breast cancer. Conclusions Because of its ease of handling, matrix-free 3D cell culture may be a useful model to study the influence of fibroblasts on breast cancer epithelial cells with use of epiFL culture medium taking this a step further towards a fully humanised 3D model. This methodology could be applied to other types of cancer cell lines, making this a versatile technique for cancer

  17. Epithelial monolayer culture system for real‐time single‐cell analyses

    PubMed Central

    Seo, Jong Bae; Moody, Mark; Koh, Duk‐Su

    2014-01-01

    Abstract Many epithelial cells form polarized monolayers under in vivo and in vitro conditions. Typically, epithelial cells are cultured for differentiation on insert systems where cells are plated on a porous filter membrane. Although the cultured monolayers have been a standard system to study epithelial physiology, there are some limits: The epithelial cells growing inside the commercial inserts are not optimal to visualize directly through lenses on inverted microscopes. The cell images are optically distorted and background fluorescence is bright due to the filter membrane positioned between the cells and the lens. In addition, the cells are not easily accessible by electrodes due to the presence of tall side walls. Here, we present the design, fabrication, and practical applications of an improved system for analysis of polarized epithelial monolayers. This new system allows (1) direct imaging of cells without an interfering filter membrane, (2) electrophysiological measurements, and (3) detection of apical secretion with minimal dilution. Therefore, our culture method is optimized to study differentiated epithelial cells at the single‐cell and subcellular levels, and can be extended to other cell types with minor modifications. PMID:24771696

  18. The effectiveness of the "Clean-Area-System" for infection control in the dental clinic.

    PubMed

    Noro, A; Suyama, Y; Takahashi, E; Chattin, B R; Hirai, Y; Takahashi, K; Ishikawa, T

    1998-02-01

    The use of effective infection control procedures and universal precautions in dental clinics, prevents cross contamination that could extend to dental health care workers and patients. The present study was initiated to investigate airborne environmental contamination in the dental clinic by viable cell count of oral streptococci grown on Mitis-Salivarius and blood agar plates. The reduction of the contamination by the "Clean-Area-System" was evaluated. "Andersen-Microbe-Sampler-Apparatus" and "Laser-Particle-Counter-System" were used for sampling and counting the bacterial cells and airborne dust, respectively. Numbers of viable cells counted as total colony forming units (CFUs) in the dental clinic were found to be significantly higher than those in the waiting room and the research laboratory. We found that the "Clean-Area-System" significantly reduced the CFUs grown on blood agar plates (p < 0.05), and that using the "Clean-Area-System" combined with the "Extra-Oral-Vacuum-Aspirator" is desirable in dental procedures such as cavity preparation. The "Extra-Oral-Vacuum-Aspirator" reduced airborne environmental contamination during tooth cutting and ultrasonic scaling procedures. In non-grinding procedures, this system proved to be very useful for infection control in the operative area. The authors concluded that the combined use of "Clean-Area-System" (dust collection ablation) and "Extra-Oral-Vacuum-Aspirator" (absorb dust ablation) was effective to reduce airborne environmental contamination in the dental clinic. We also fully confirmed that oral streptococci were an adequate indicator in the assessment for infection control in dental institutions. PMID:9663027

  19. Effect of agitation and terminal subcultures on yield and speed of detection of the Oxoid Signal blood culture system versus the BACTEC radiometric system

    SciTech Connect

    Weinstein, M.P.; Mirrett, S.; Reimer, L.G.; Reller, L.B.

    1989-03-01

    In an initial evaluation, we found the Oxoid Signal blood culture system inferior to the BACTEC radiometric system for detection of some microorganisms causing septicemia. To determine whether modified processing of the Oxoid Signal blood culture system could improve its yield and speed of detecting positive cultures relative to the BACTEC radiometric system, we agitated all Oxoid bottles during the first 24 to 48 h of incubation and performed aerobic and anaerobic subcultures of all Oxoid bottles negative after 7 days of incubation. These modifications improved the overall performance of the Oxoid system, particularly with regard to the yield of streptococci, members of the family Enterobacteriaceae, and Haemophilus, Neisseria, and Acinetobacter spp. The speed of detecting positive cultures also was improved, especially within the first 24 h of incubation. However, the BACTEC system still detected more positive cultures (P less than 0.005), especially of obligate aerobes such as Pseudomonas aeruginosa (P less than 0.05) and yeasts (P less than 0.005). The BACTEC system also detected positive cultures earlier than the Oxoid system (e.g., at 24 h of incubation, 70.5% of BACTEC positive cultures detected versus 62.1% of Oxoid positive cultures detected). Further modifications of the Oxoid system which might include a revised medium, additional processing modifications, altered headspace atmosphere, or a complementary second broth medium should be considered, since the system is attractive in concept and is easy to use in the clinical laboratory.

  20. Hyperspectral image acquisition and analysis of cultured bacteria for the discrimination of urinary tract infections.

    PubMed

    Turra, Giovanni; Conti, Nicola; Signoroni, Alberto

    2015-08-01

    Because of their widespread diffusion and impact on human health, early identification of pathogens responsible for urinary tract infections (UTI) is one of the main challenges of clinical microbiology. Currently, bacteria culturing on Chromogenic plates is widely adopted for UTI detection for its readily interpretable visual outcomes. However, the search of alternate solutions can be highly attractive, especially in the rapidly developing context of bacteriology laboratory automation and digitization, as long as they can improve cost-effectiveness or allow early discrimination. In this work, we consider and develop hyperspectral image acquisition and analysis solutions to verify the feasibility of a "virtual chromogenic agar" approach, based on the acquisition of spectral signatures from bacterial colonies growing on blood agar plates, and their interpretation by means of machine learning solutions. We implemented and tested two classification approaches (PCA+SVM and RSIMCA) that evidenced good capability to discriminate among five selected UTI bacteria. For its better performance, robustness and attitude to work with an expanding set of pathogens, we conclude that the RSIMCA-based approach is worth to be further investigated in a clinical usage perspective. PMID:26736373

  1. Three-dimensional culture systems in cancer research: Focus on tumor spheroid model.

    PubMed

    Nath, Sritama; Devi, Gayathri R

    2016-07-01

    Cancer cells propagated in three-dimensional (3D) culture systems exhibit physiologically relevant cell-cell and cell-matrix interactions, gene expression and signaling pathway profiles, heterogeneity and structural complexity that reflect in vivo tumors. In recent years, development of various 3D models has improved the study of host-tumor interaction and use of high-throughput screening platforms for anti-cancer drug discovery and development. This review attempts to summarize the various 3D culture systems, with an emphasis on the most well characterized and widely applied model - multicellular tumor spheroids. This review also highlights the various techniques to generate tumor spheroids, methods to characterize them, and its applicability in cancer research. PMID:27063403

  2. Recent Progress in Endothelial Progenitor Cell Culture Systems: Potential for Stroke Therapy

    PubMed Central

    TAKIZAWA, Shunya; NAGATA, Eiichiro; NAKAYAMA, Taira; MASUDA, Haruchika; ASAHARA, Takayuki

    2016-01-01

    Endothelial progenitor cells (EPCs) participate in endothelial repair and angiogenesis due to their abilities to differentiate into endothelial cells and to secrete protective cytokines and growth factors. Consequently, there is considerable interest in cell therapy with EPCs isolated from peripheral blood to treat various ischemic injuries. Quality and quantity-controlled culture systems to obtain mononuclear cells enriched in EPCs with well-defined angiogenic and anti-inflammatory phenotypes have recently been developed, and increasing evidence from animal models and clinical trials supports the idea that transplantation of EPCs contributes to the regenerative process in ischemic organs and is effective for the therapy of ischemic cerebral injury. Here, we briefly describe the general characteristics of EPCs, and we review recent developments in culture systems and applications of EPCs and EPC-enriched cell populations to treat ischemic stroke. PMID:27041632

  3. [A study of culture-based easy identification system for Malassezia].

    PubMed

    Kaneko, Takamasa

    2011-01-01

    Most species of this genus are lipid-dependent yeasts, which colonize the seborrheic part of the skin, and they have been reported to be associated with pityriasis versicolor, Malassezia folliculitis, seborrheic dermatitis, and atopic dermatitis. Malassezia have been re-classified into 7 species based on molecular biological analysis of nuclear ribosomal DNA/RNA and new Malassezia species were reported. As members of the genus Malassezia share similar morphological and biochemical characteristics, it was thought to be difficult to differentiate between them based on phenotypic features. While molecular biological techniques are the most reliable methods for identification of Malassezia, they are not available in most clinical laboratories. We studied ( i ) development of an efficient isolation media and culture based easy identification system, ( ii ) the incidence of atypical biochemical features in Malassezia species and propose a culture-based easy identification system for clinically important Malassezia species, M. globosa, M. restricta, and M. furfur. PMID:22123328

  4. Plasma-Mediated Inactivation of Pseudomonas aeruginosa Biofilms Grown on Borosilicate Surfaces under Continuous Culture System

    PubMed Central

    Vandervoort, Kurt G.; Brelles-Mariño, Graciela

    2014-01-01

    Biofilms are microbial communities attached to a surface and embedded in a matrix composed of exopolysaccharides and excreted nucleic acids. Bacterial biofilms are responsible for undesirable effects such as disease, prostheses colonization, biofouling, equipment damage, and pipe plugging. Biofilms are also more resilient than free-living cells to regular sterilization methods and therefore it is indispensable to develop better ways to control and remove them. The use of gas discharge plasmas is a good alternative since plasmas contain a mixture of reactive agents well-known for their decontamination potential against free microorganisms. We have previously reported that Pseudomonas aeruginosa biofilms were inactivated after a 1-min plasma exposure. We determined that the adhesiveness and the thickness of Pseudomonas biofilms grown on borosilicate were reduced. We also reported sequential morphological changes and loss of viability upon plasma treatment. However, the studies were carried out in batch cultures. The use of a continuous culture results in a more homogenous environment ensuring reproducible biofilm growth. The aim of this work was to study plasma-mediated inactivation of P. aeruginosa biofilms grown on borosilicate in a continuous culture system. In this paper we show that biofilms grown on glass under continuous culture can be inactivated by using gas discharge plasma. Both biofilm architecture and cell culturabilty are impacted by the plasma treatment. The inactivation kinetics is similar to previously described ones and cells go through sequential changes ranging from minimal modification without loss of viability at short plasma exposure times, to major structure and viability loss at longer exposure times. We report that changes in biofilm structure leading to the loss of culturability and viability are related to a decrease of the biofilm matrix adhesiveness. To our knowledge, there has been no attempt to evaluate the inactivation

  5. Single-Plasmid-Based System for Efficient Noncanonical Amino Acid Mutagenesis in Cultured Mammalian Cells.

    PubMed

    Cohen, Sarit; Arbely, Eyal

    2016-06-01

    We describe a new expression system for efficient non-canonical amino acid mutagenesis in cultured mammalian cells by using the pyrrolysine tRNA synthetase/tRNACUA (Pyl) pair. A significant improvement in the incorporation of non-canonical amino acids into proteins was obtained by combining all the required genetic components into a single and compact vector that can be efficiently delivered to different mammalian cell lines by conventional transfection reagents. PMID:27120490

  6. Utilizing a Robotic Sprayer for High Lateral and Mass Resolution MALDI FT-ICR MSI of Microbial Cultures

    NASA Astrophysics Data System (ADS)

    Anderton, Christopher R.; Chu, Rosalie K.; Tolić, Nikola; Creissen, Alain; Paša-Tolić, Ljiljana

    2016-03-01

    The ability to visualize biochemical interactions between microbial communities using MALDI MSI has provided tremendous insights into a variety of biological fields. Matrix application using a sieve proved to be incredibly useful, but it has many limitations that include uneven matrix coverage and limitation in the types of matrices that could be employed in studies. Recently, there has been a concerted effort to improve matrix application for studying agar plated microbial cultures, many of which utilized automated matrix sprayers. Here, we describe the usefulness of using a robotic sprayer for matrix application. The robotic sprayer has two-dimensional control over where matrix is applied, and a heated capillary that allows for rapid drying of the applied matrix. This method provided a significant increase in MALDI sensitivity over the sieve method, as demonstrated by FT-ICR MS analysis, facilitating the ability to gain higher lateral resolution MS images of Bacillus subtilis than previously reported. This method also allowed for the use of different matrices to be applied to the culture surfaces.

  7. Tailoring microfluidic systems for organ-like cell culture applications using multiphysics simulations

    NASA Astrophysics Data System (ADS)

    Hagmeyer, Britta; Schütte, Julia; Böttger, Jan; Gebhardt, Rolf; Stelzle, Martin

    2013-03-01

    Replacing animal testing with in vitro cocultures of human cells is a long-term goal in pre-clinical drug tests used to gain reliable insight into drug-induced cell toxicity. However, current state-of-the-art 2D or 3D cell cultures aiming at mimicking human organs in vitro still lack organ-like morphology and perfusion and thus organ-like functions. To this end, microfluidic systems enable construction of cell culture devices which can be designed to more closely resemble the smallest functional unit of organs. Multiphysics simulations represent a powerful tool to study the various relevant physical phenomena and their impact on functionality inside microfluidic structures. This is particularly useful as it allows for assessment of system functions already during the design stage prior to actual chip fabrication. In the HepaChip®, dielectrophoretic forces are used to assemble human hepatocytes and human endothelial cells in liver sinusoid-like structures. Numerical simulations of flow distribution, shear stress, electrical fields and heat dissipation inside the cell assembly chambers as well as surface wetting and surface tension effects during filling of the microchannel network supported the design of this human-liver-on-chip microfluidic system for cell culture applications. Based on the device design resulting thereof, a prototype chip was injection-moulded in COP (cyclic olefin polymer). Functional hepatocyte and endothelial cell cocultures were established inside the HepaChip® showing excellent metabolic and secretory performance.

  8. Biodegradation of Endocrine Disruptors in Solid-Liquid Two-Phase Partitioning Systems by Enrichment Cultures

    PubMed Central

    dos Santos, Silvia Cristina Cunha; Ouellette, Julianne; Juteau, Pierre; Lépine, François; Déziel, Eric

    2013-01-01

    Naturally occurring and synthetic estrogens and other molecules from industrial sources strongly contribute to the endocrine disruption of urban wastewater. Because of the presence of these molecules in low but effective concentrations in wastewaters, these endocrine disruptors (EDs) are only partially removed after most wastewater treatments, reflecting the presence of these molecules in rivers in urban areas. The development of a two-phase partitioning bioreactor (TPPB) might be an effective strategy for the removal of EDs from wastewater plant effluents. Here, we describe the establishment of three ED-degrading microbial enrichment cultures adapted to a solid-liquid two-phase partitioning system using Hytrel as the immiscible water phase and loaded with estrone, estradiol, estriol, ethynylestradiol, nonylphenol, and bisphenol A. All molecules except ethynylestradiol were degraded in the enrichment cultures. The bacterial composition of the three enrichment cultures was determined using 16S rRNA gene sequencing and showed sequences affiliated with bacteria associated with the degradation of these compounds, such as Sphingomonadales. One Rhodococcus isolate capable of degrading estrone, estradiol, and estriol was isolated from one enrichment culture. These results highlight the great potential for the development of TPPB for the degradation of highly diluted EDs in water effluents. PMID:23728808

  9. Characteristics of human dendritic cells generated in a microgravity analog culture system

    NASA Technical Reports Server (NTRS)

    Savary, C. A.; Grazziuti, M. L.; Przepiorka, D.; Tomasovic, S. P.; McIntyre, B. W.; Woodside, D. G.; Pellis, N. R.; Pierson, D. L.; Rex, J. H.; McIntire, L. V. (Principal Investigator)

    2001-01-01

    Generation of an effective immune response requires that antigens be processed and presented to T lymphocytes by antigen-presenting cells, the most efficient of which are dendritic cells (DC). Because of their influence on both the innate and the acquired arms of immunity, a defect in DC would be expected to result in a broad impairment of immune function, not unlike that observed in astronauts during or after space flight. In the study reported here, we investigated whether DC generation and function are altered in a culture environment that models microgravity, i.e., the rotary-cell culture system (RCCS). We observed that RCCS supported the generation of DC identified by morphology, phenotype (HLA-DR+ and lacking lineage-associated markers), and function (high allostimulatory activity). However, the yield of DC from RCCS was significantly lower than that from static cultures. RCCS-generated DC were less able to phagocytose Aspergillus fumigatus conidia and expressed a lower density of surface HLA-DR. The proportion of DC expressing CD80 was also significantly reduced in RCCS compared to static cultures. When exposed to fungal antigens, RCCS-generated DC produced lower levels of interleukin-12 and failed to upregulate some costimulatory/adhesion molecules involved in antigen presentation. These data suggest that DC generation, and some functions needed to mount an effective immune response to pathogens, may be disturbed in the microgravity environment of space.

  10. Probing Hypoxia-Induced Staurosporine Resistance in Prostate Cancer Cells with a Microfluidic Culture System

    PubMed Central

    Khanal, Grishma; Hiemstra, Scott

    2014-01-01

    A microfluidic system for cell culture and drug response studies was developed to elucidate the effects of hypoxia on drug susceptibility. Drug response studies were performed in prostate cancer cells and Ramos B cells under normoxic and hypoxic conditions. A vacuum actuated microfluidic culture device was used for cell culture and PC3 cells were cultured in the chip up to 16 hours. Cells were treated with several concentrations of staurosporine and apoptosis was assayed using the fluorescent probes MitoTracker Red and Annexin-V. For hypoxic samples, the chip was placed in a hypoxia chamber and pre-conditioned at <1% oxygen before inducing the cells with staurosporine. Cells exposed to 2 μM staurosporine were 32% ± 10% apoptotic under normoxic conditions but only 1.5% ± 12% apoptotic under hypoxic conditions. As little as 1 hour of hypoxic preconditioning increased drug resistance. Cell apoptosis correlated with drug dose, although in each case hypoxia reduced the apoptotic fraction significantly. Given the rapid nature of cell adaptation to hypoxia, this chip and analysis approach can be used to identify compounds that can induce cell death in hypoxic tumor cells rapidly. PMID:24479128

  11. Enhancement of antibody production by growth factor addition in perfusion and hollow-fiber culture systems.

    PubMed

    Omasa, T; Kobayashi, M; Nishikawa, T; Shioya, S; Suga, K; Uemura, S; Kitani, Y; Imamura, Y

    1995-12-20

    The effects of the high-molecular-weight growth factors, transferrin and bovine serum albumin (BSA), on antibody production were analyzed quantitatively in continuous hollow-fiber cultivation over a period of 60 days. Transferrin enhanced cell growth but had no significant effect on the specific antibody production rate, whereas BSA significantly enhanced antibody production. The antibody production rate was increased 4- and 14-fold respectively by feeding BSA at 2 and 5 g L(-1) into the EC side of the system (the side connected to the cell-containing outer part of the hollow-fiber unit) compared with the production achieved without BSA. Addition of 5 g L(1) BSA into the IC side of the system (the side connected to the inner part of the hollow-fiber unit) resulted in a 2.5-fold increase in the antibody production rate. The effect of BSA was also analyzed using the perfusion culture system with a separation unit. When fresh medium containing either 2 or 5 g L(-1) BSA was fed into the reactor, both the specific growth rate and specific death rate increased, while the specific antibody production rate was increased 2- and 25-fold, respectively, by feeding BSA at these two concentrations compared with no addition. Comparing the two systems, the increase in the antibody production rate achieved with the hollow-fiber system was threefold greater than that in the perfusion culture system with the same concentration of BSA feeding. (c) 1995 John Wiley & Sons, Inc. PMID:18623537

  12. Zika Virus Infectious Cell Culture System and the In Vitro Prophylactic Effect of Interferons.

    PubMed

    Contreras, Deisy; Arumugaswami, Vaithilingaraja

    2016-01-01

    Zika Virus (ZIKV) is an emerging pathogen that is linked to fetal developmental abnormalities such as microcephaly, eye defects, and impaired growth. ZIKV is an RNA virus of the Flaviviridae family. ZIKV is mainly transmitted by mosquitoes, but can also be spread by maternal to fetal vertical transmission as well as sexual contact. To date, there are no reliable treatment or vaccine options available to protect those infected by the virus. The development of a reproducible, effective Zika virus infectious cell culture system is critical for studying the molecular mechanisms of ZIKV replication as well as drug and vaccine development. In this regard, a protocol describing a mammalian cell-based in vitro Zika virus culture system for viral production and growth analysis is reported here. Details on the formation of plaques by Zika virus on a cell monolayer and plaque assay for measuring viral titer are presented. Viral genome replication kinetics and double-stranded RNA genome replicatory intermediates are determined. This culture platform was utilized to screen against a library of a small set of cytokines resulting in the identification of interferon-α (IFN-α), IFN-β and IFN-γ as potent inhibitors of Zika viral growth. In summary, an in vitro infectious Zika viral culture system and various virological assays are demonstrated in this study, which has the potential to greatly benefit the research community in elucidating further the mechanisms of viral pathogenesis and the evolution of viral virulence. Antiviral IFN-alpha can further be evaluated as a prophylactic, post-exposure prophylactic, and treatment option for Zika virus infections in high-risk populations, including infected pregnant women. PMID:27584546

  13. Development and characterization of cell culture systems from Puntius (Tor) chelynoides (McClelland).

    PubMed

    Goswami, M; Sharma, B S; Tripathi, A K; Yadav, Kamalendra; Bahuguna, S N; Nagpure, N S; Lakra, W S; Jena, J K

    2012-05-25

    Puntius (Tor) chelynoides, commonly known as dark mahseer, is a commercially important coldwater fish species which inhabits fast-flowing hill-streams of India and Nepal. Cell culture systems were developed from eye, fin, heart and swim bladder tissues of P. chelynoides using explant method. The cell culture system developed from eye has been maintained towards a continuous cell line designated as PCE. The cells were grown in 25cm(2) tissue culture flasks with Leibovitz' L-15 media supplemented with 20 % fetal bovine serum (FBS) at 24°C. The PCE cell line consists of predominantly fibroblast-like cells and showed high plating efficiency. The monolayer formed from the fin and heart explants were comprised of epithelial as well as fibroblast-like cells, a prominent and rhythmic heartbeat was also observed in heart explants. Monolayer formed from swim bladder explants showed the morphology of fibroblast-like cells. All the cells from different tissues are able to grow at an optimum temperature of 24°C and growth rate increased as the FBS concentration increased. The PCE cell line was characterized using amplification of mitochondrial cytochrome oxidase subunit I (COI) & 16S rRNA genes which confirmed that the cell line originated from P. chelynoides. Cytogenetic analysis of PCE cell line and cells from fin revealed a diploid count of 100 chromosomes. Upon transfection with pEGFP-C1 plasmid, bright fluorescent signals were observed, suggesting that this cell line can be used for transgenic and genetic manipulation studies. Further, genotoxicity assessment of PCE cells illustrated the utility of this cell line as an in vitro model for aquatic toxicological studies. The PCE cell line was successfully cryopreserved and revived at different passage levels. The cell line and culture systems are being maintained to develop continuous cell lines for further studies. PMID:22465535

  14. Using Remote Sensing Technology on the Delimitation of the Conservation Area for the Jianan Irrigation System Cultural Landsccape

    NASA Astrophysics Data System (ADS)

    Wang, C. H.

    2015-08-01

    In recent years the cultural landscape has become an important issue for cultural heritages throughout the world. It represents the "combined works of nature and of man" designated in Article 1 of the World Heritage Convention. When a landscape has a cultural heritage value, important features should be marked and mapped through the delimitation of a conservation area, which may be essential for further conservation work. However, a cultural landscape's spatial area is usually wider than the ordinary architectural type of cultural heritage, since various elements and impact factors, forming the cultural landscape's character, lie within a wide geographic area. It is argued that the conservation of a cultural landscape may be influenced by the delimitation of the conservation area, the corresponding land management measures, the limits and encouragements. The Jianan Irrigation System, an historical cultural landscape in southern Taiwan, was registered as a living cultural heritage site in 2009. However, the system's conservation should not be limited to just only the reservoir or canals, but expanded to irrigated areas where farmland may be the most relevant. Through the analysis process, only approximately 42,000 hectares was defined as a conservation area, but closely related to agricultural plantations and irrigated by the system. This is only half of the 1977 irrigated area due to urban sprawl and continuous industrial expansion.

  15. Simplified three-dimensional culture system for long-term expansion of embryonic stem cells

    PubMed Central

    McKee, Christina; Perez-Cruet, Mick; Chavez, Ferman; Chaudhry, G Rasul

    2015-01-01

    proliferation as determined by confocal microscopy and PB staining. 3-D scaffold cultured ESCs expressed significantly higher levels (P < 0.01) of Oct4, Nanog, and Kl4, showing a 2.8, 3.0 and 1.8 fold increase, respectively, in comparison to 2-D grown cells. A similar increase in the protein expression levels of Oct4, Nanog, and Klf4 was observed in 3-D grown ESCs. However, when 3-D cultured ESCs were subsequently passaged in 2-D culture conditions, the level of these pluripotent markers was reduced to normal levels. 3-D grown ESCs produced teratomas and yielded cells of all three germ layers, expressing brachyury (mesoderm), NCAM (ectoderm), and GATA4 (endoderm) markers. Furthermore, these cells differentiated into osteogenic, chondrogenic, myogenic, and neural lineages expressing Col1, Col2, Myog, and Nestin, respectively. CONCLUSION: This novel 3-D culture system demonstrated long-term maintenance of mouse ESCs without the routine passaging and manipulation necessary for traditional 2-D cell propagation. PMID:26328022

  16. Numerical calculations for diffusion effects in the well-of-the-well culture system for mammalian embryos.

    PubMed

    Matsuura, Koji

    2014-06-01

    Recent studies suggest that the microenvironment and embryo density used during embryo culture considerably affect development to the blastocyst stage. High embryo density allows for autocrine secretions to diffuse to neighbouring embryos during group culture, with a positive effect on further development. A variation of group culture is the well-of-the-well (WOW) culture system, allowing for individual identification of embryos cultured in small holes in a microdroplet. Bovine blastocyst development is higher in the WOW culture system than in conventional group culture. To compare the concentration of chemical factors between conventional and WOW culture, a model was constructed to calculate the concentration of secreted factors based on Fick's second law of diffusion using spreadsheet software. Furthermore, model was used to determine the concentration of growth factors and waste materials adjacent to the embryo periphery. The results of these calculations suggest that the highest difference in the concentration of secreted small molecules and macromolecules was at the most two- to threefold, with the concentrations reduced more and diffusion kinetics facilitated to a greater extent in the WOW culture system. The average ratio of the concentration of secreted macromolecules (10nm diameter) around the embryos was also compared between systems with well widths of 0.1 and 0.3mm. The concentration of secreted materials surrounding embryos increased in a narrow tapered well. The findings suggest that the WOW culture system is better than conventional group culture because of the increased final concentration of autocrine factors and higher diffusion kinetics of waste materials. PMID:23697480

  17. Psychosocial and Cultural Modeling in Human Computation Systems: A Gamification Approach

    SciTech Connect

    Sanfilippo, Antonio P.; Riensche, Roderick M.; Haack, Jereme N.; Butner, R. Scott

    2013-11-20

    “Gamification”, the application of gameplay to real-world problems, enables the development of human computation systems that support decision-making through the integration of social and machine intelligence. One of gamification’s major benefits includes the creation of a problem solving environment where the influence of cognitive and cultural biases on human judgment can be curtailed through collaborative and competitive reasoning. By reducing biases on human judgment, gamification allows human computation systems to exploit human creativity relatively unhindered by human error. Operationally, gamification uses simulation to harvest human behavioral data that provide valuable insights for the solution of real-world problems.

  18. Copyright for Interactive Systems: Stratagems for Tourism and Cultural Heritage Promotion

    NASA Astrophysics Data System (ADS)

    Cipolla-Ficarra, Francisco V.; Cipolla-Ficarra, Miguel; Ficarra, Valeria M.

    We present a series of strategies followed from the interactive design for the realization of a hypermedia system aimed at promoting in an original, simple and universal way the cultural and tourism heritage of a wide rural area in two Italian regions: Emilia Romagna and Lombardy. Besides, the main stratagems followed are disclosed to overcome the existing hurdles when it comes to copyright for the free diffusion of the tourism view of the area, such as can be photography or video, for instance. Finally, we present the first vademecum to be considered before making on-line and off-line interactive systems in Italy.

  19. Bridging Diversity and Family Systems: Culturally Informed and Flexible Family Based Treatment for Hispanic Adolescents

    PubMed Central

    Santisteban, Daniel A.; Mena, Maite P.; Abalo, Clara

    2014-01-01

    There is growing interest in identifying interventions that have been tested and found efficacious with minority families. This interest is fueled in part by the growth of Hispanics in the U.S. as well as by research findings that suggest that Hispanics have better outcomes when treatments are adapted to their unique experiences, and risk and protective factors. Family-based treatments for culturally diverse populations require the integration of advances from both the cultural and family systems domains. Current intervention research has begun to move towards developing and advancing individualized interventions for patients/clients. Adaptive interventions, tailored interventions, adapted interventions, and targeted interventions have all been identified in the literature as appropriate for addressing distinct cultural characteristics which generic interventions may not address effectively. To date, research has focused less on tailored or adaptive interventions partly due to the fact that they require decision rules, more careful implementation, and measurement of individualized outcomes. In this article we present evidence for the usefulness of adaptive interventions that can address not only subgroup variability but within group variability as well. Culturally Informed and Flexible Family-Based Treatment for Adolescents is presented as an adaptive treatment that allows for the tailoring of treatment to the unique clinical and cultural variations of individual adolescents and families, but that does so in a systematic and replicable fashion. By building decision-making processes into the manualized treatment, the transportability of the treatment may be enhanced as family therapists appreciate it’s flexibility to address the complexity of clinical work. PMID:24772378

  20. Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors

    PubMed Central

    Edmondson, Rasheena; Broglie, Jessica Jenkins; Adcock, Audrey F.

    2014-01-01

    Abstract Three-dimensional (3D) cell culture systems have gained increasing interest in drug discovery and tissue engineering due to their evident advantages in providing more physiologically relevant information and more predictive data for in vivo tests. In this review, we discuss the characteristics of 3D cell culture systems in comparison to the two-dimensional (2D) monolayer culture, focusing on cell growth conditions, cell proliferation, population, and gene and protein expression profiles. The innovations and development in 3D culture systems for drug discovery over the past 5 years are also reviewed in the article, emphasizing the cellular response to different classes of anticancer drugs, focusing particularly on similarities and differences between 3D and 2D models across the field. The progression and advancement in the application of 3D cell cultures in cell-based biosensors is another focal point of this review. PMID:24831787

  1. Viral Load Analysis of Hepatitis C Virus in Huh7.5 Cell Culture System

    PubMed Central

    Teimourpour, Roghayeh; Meshkat, Zahra; Gholoubi, Aida; Nomani, Hosein; Rostami, Sina

    2015-01-01

    Background: Previous studies using cell culture systems for the replication of hepatitis C virus have opened new research dimensions, and paved the ways for further and detailed studies of the virus in vitro. Objectives: The purpose of the present study was to cultivate hepatitis C virus in a cell culture system and evaluate viral amplification. Materials and Methods: In order to propagate hepatitis C virus, cloned whole genome of virus, JFH-1, was used. JFH-1 cDNA was introduced into strain JM109 of Escherichia coli and plasmid, containing the viral genome was purified from transformed bacteria. After XbaI digestion, RNA synthesis was induced using T7 RNA polymerase enzyme. Next, eukaryotic cell line Huh 7.5 was transfected by the purified RNA. Finally, Huh-7.5 cell line was infected with replicated virus and viral load was determined using real-time PCR (Polymerase Chain Reaction). Results: The amount of viral load, which was measured using real-time PCR was 17592 IU/mL. Conclusions: In the present study, using cell culture, a high titer (in acceptable range) of infectious hepatitis C virus was produced. This method could be used in future studies. PMID:26290686

  2. Evaluation of human hepatocytes cultured by three-dimensional spheroid systems for drug metabolism.

    PubMed

    Ohkura, Takako; Ohta, Kunihiro; Nagao, Takuya; Kusumoto, Kumiko; Koeda, Akiko; Ueda, Tadayoshi; Jomura, Tomoko; Ikeya, Takeshi; Ozeki, Emiko; Wada, Kazuki; Naitoh, Kazushi; Inoue, Yukiko; Takahashi, Naoki; Iwai, Hisakazu; Arakawa, Hiroshi; Ogihara, Takuo

    2014-01-01

    We investigated the utility of three-dimensional (3D) spheroid cultures of human hepatocytes in discovering drug metabolites. Metabolites of acetaminophen, diclofenac, lamotrigine, midazolam, propranolol and salbutamol were analyzed by liquid chromatography-tandem mass spectrometry (LC/MS/MS) to measure enzyme activities in this system cultured for 2 and 7 days. Sequential metabolic reactions by Phase I and then Phase II enzymes were found in diclofenac [CYP2C9 and UDP-glucuronyltransferases (UGTs)], midazolam (CYP3A4 and UGTs) and propranolol (CYP1A2/2D6 and UGTs). Moreover, lamotrigine and salbutamol were metabolized to lamotrigine-N-glucuronide and salbutamol 4-O-sulfate, respectively. These metabolites, which are human specific, could be observed in clinical studies, but not in conventional hepatic culture systems as in previous reports. Acetaminophen was metabolized to glucuronide and sulfate conjugates, and N-acetyl-p-benzo-quinoneimine (NAPQI) and its metabolites were not observed. In addition, mRNA of drug-metabolism enzymes [CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, UGT1A1, UGT2B7, sulfotransferase 1A1 (SULT1A1) and glutathione S-transferase pi 1 (GSTP1)], which were measured by qRT-PCR, were expressed in the human hepatocyte spheroids. In conclusion, these results suggest that human hepatocyte spheroids are useful in discovering drug metabolites. PMID:24695277

  3. A portable immunomagnetic cell capture system to accelerate culture diagnosis of bacterial infections.

    PubMed

    Singh, Saurabh; Upadhyay, Mohita; Sharma, Jyoti; Gupta, Shalini; Vivekanandan, Perumal; Elangovan, Ravikrishnan

    2016-05-23

    Bacterial infections continue to be a major cause of deaths globally, particularly in resource-poor settings. In the absence of rapid and affordable diagnostic solutions, patients are mostly administered broad spectrum antibiotics leading to antibiotics resistance and poor recovery. Culture diagnosis continues to be a gold standard for diagnosis of bacterial infection, despite its long turnaround time of 24 to 48 h. We have developed a portable immunomagnetic cell capture (iMC(2)) system that allows rapid culture diagnosis of bacterial pathogens. Our approach involves the culture growth of the blood samples in broth media for 6 to 8 h, followed by immunomagnetic enrichment of the target cells using the iMC(2) device. The device comprises a disposable capture chip that has two chambers of 5 ml and 50 μl volume connected through a channel with a manual valve. Bacterial cells bound to antibody coated magnetic nanoparticles are swept from the 5 ml sample chamber into the 50 μl recovery chamber by moving an external magnetic field with respect to the capture chip using a linear positioner. This enables specific isolation and up to 100× enrichment of the target cells. The presence of bacteria in the recovered sample is confirmed visually using a lateral flow immunoassay. The system is demonstrated in buffer and blood samples spiked with S. typhi. The method has high sensitivity (10 CFU ml(-1)), specificity and a rapid turnaround time of less than 7 h, a significant improvement over conventional methods. PMID:27118505

  4. Understanding the earth systems of Malawi: Ecological sustainability, culture, and place-based education

    NASA Astrophysics Data System (ADS)

    Glasson, George E.; Frykholm, Jeffrey A.; Mhango, Ndalapa A.; Phiri, Absalom D.

    2006-07-01

    The purpose of this 2-year study was to investigate Malawian teacher educators' perspectives and dispositions toward teaching about ecological sustainability issues in Malawi, a developing country in sub-Sahara Africa. This study was embedded in a larger theoretical framework of investigating earth systems science through the understanding of nature-knowledge-culture systems from local, place-based perspectives. Specifically, we were interested in learning more about eco-justice issues that are related to environmental degradation in Malawi and the potential role of inquiry-oriented pedagogies in addressing these issues. In a science methods course, the African educators' views on deforestation and teaching about ecological sustainability were explored within the context of the local environment and culture. Teachers participated in inquiry pedagogies designed to promote the sharing of perspectives related to the connections between culture and ecological degradation. Strategies encouraging dialogue and reflection included role-playing, class discussions, curriculum development activities, teaching experiences with children, and field trips to a nature preserve. Data were analyzed from postcolonial and critical pedagogy of place theoretical perspectives to better understand the hybridization of viewpoints influenced by both Western and indigenous science and the political hegemonies that impact sustainable living in Malawi. Findings suggested that the colonial legacy of Malawi continues to impact the ecological sustainability issue of deforestation. Inquiry-oriented pedagogies and connections to indigenous science were embraced by the Malawian educators as a means to involve children in investigation, decision making, and ownership of critical environmental issues.

  5. Effect of different culture systems on adipocyte differentiation-related protein (ADRP) in bovine embryos.

    PubMed

    Al Darwich, A; Perreau, C; Tsikis, G; Coudert, E; Touzé, J L; Briant, E; Beckers, J F; Mermillod, P; Guignot, F

    2014-03-01

    Bovine embryos cultured in serum-containing media abnormally accumulate lipid droplets, compared to their in vivo counterparts. The objective of this study was to investigate the effect of different culture systems on the mRNA expression and on the quantification and localisation of adipocyte differentiation-related protein (ADRP), a protein associated with lipid accumulation in bovine blastocysts. Two experiments were independently performed for ADRP mRNA expression analysis. In experiment A, blastocysts were produced in modified synthetic oviduct fluid (mSOF)+10% foetal calf serum (FCS), in coculture (bovine oviduct epithelial cells, Boec) and in ewe oviducts, whereas in experiment B, they were produced in mSOF+10μM docosahexaenoic acid (DHA) and in vivo. Control groups were also performed. ADRP mRNA expression was downregulated in the Boec, ewe oviduct and in vivo groups compared to the 10% FCS or DHA groups, respectively. Moreover, the expression of this protein was downregulated in the Boec group compared to the control group (P<0.05). A third experiment (experiment C) was performed to quantify and localise ADRP protein. Boec, in vivo and control groups were tested. After immunofluorescence staining followed by confocal microscopy analysis, embryonic ADRP was clearly localised around lipid droplets, indicating that ADRP is also a lipid droplet coat protein in bovine embryos. In conclusion, our results demonstrate that bovine embryos at the blastocyst stage expressed ADRP mRNA and protein, and that the embryonic culture system modified this expression. PMID:24560670

  6. A two dimensional clinostat experiment for microalgae cultures - basic work for bio- regenerativ life support systems

    NASA Astrophysics Data System (ADS)

    Harting, Benjamin; Slenzka, Klaus

    2012-07-01

    To investigate the influence of microgravity environments on photosynthetic organisms we designed a 2 dimensional clinostatexperiment for a suspended cell culture of Chlamydomonas reinhardtii. A novel approach of online measurments concerning relevant parameters important for the clasification of photosynthesis was obtained. To adress the photosynthesis rate we installed and validated an optical mesurement system to monitor the evolution and consumption of dissolved oxygen. Simultaneously a PAM sensor to analyse the flourescence quantum yield of the photochemical reaction was integarted. Thus it was possible to directly classify important parameters of the phototrophic metabolism during clinorotation. The experiment design including well suited light conditions and further biochemical analysis were directly performed for microalgal cell cultures. Changes in the photosynthetic efficiancy of phototrophic cyanobacteria has been observed during parabolic flight campaign but the cause is already not understood. Explenations could be the dependency of gravitaxis by intracellular ionconcentartion or the existance of mechanosensitive ionchannels for example associated in chloroplasts of Chlamydomonas reinhardtii. The purpuse of the microalgal clinostat are studies in a qasi microgravity environment for the process design of future bioregenerative life suport systems in spaceflight missions. First results has indicated the need for special nourishment of the cell culture during microgravity experiments. Further data will be presented during the assembly.

  7. Research and development of shallow algal mass culture systems for the production of oils

    SciTech Connect

    Laws, E.A.

    1984-10-01

    The major accomplishment of the past nine months' work was the identification of a microalgal species which can be grown in the system on a 12-month basis without temperature control. The most promising species identified to date is a strain of platymonas sp. This strain grows rapidly at temperatures from 20/sup 0/ to 34/sup 0/C, and at salinities from 1.5 to 3.5%. Neither the lower temperature limit nor the lower salinity limit of the strain are known at this time. A factorial experiment designed to determine optimum growth conditions indicated that the optimum culture depth was 10 cm, the optimum pH about 7.5, and the optimum flow rate about 30 cm/s. A major discovery was that diluting the culture every third day greatly enhanced production. In this dilution mode daily yields averaged 46 g/m/sup 2/ ash-free dry weight (AFDW) over a one-month period, and photosynthetic efficiencies averaged 11% (based on visible light energy). The former figure is over twice the best long-term yields achieved in microalgal mass culture systems grown exclusively on inorganic nutrients.

  8. Computer-controlled microcirculatory support system for endothelial cell culture and shearing.

    PubMed

    Song, Jonathan W; Gu, Wei; Futai, Nobuyuki; Warner, Kristy A; Nor, Jacques E; Takayama, Shuichi

    2005-07-01

    Endothelial cells (ECs) lining the inner lumen of blood vessels are continuously subjected to hemodynamic shear stress, which is known to modify EC morphology and biological activity. This paper describes a self-contained microcirculatory EC culture system that efficiently studies such effects of shear stress on EC alignment and elongation in vitro. The culture system is composed of elastomeric microfluidic cell shearing chambers interfaced with computer-controlled movement of piezoelectric pins on a refreshable Braille display. The flow rate is varied by design of channels that allow for movement of different volumes of fluid per variable-speed pump stroke. The integrated microfluidic valving and pumping system allowed primary EC seeding and differential shearing in multiple compartments to be performed on a single chip. The microfluidic flows caused ECs to align and elongate significantly in the direction of flow according to their exposed levels of shear stress. This microfluidic system overcomes the small flow rates and the inefficiencies of previously described microfluidic and macroscopic systems respectively to conveniently perform parallel studies of EC response to shear stress. PMID:15987102

  9. A Novel Feeder-Free Culture System for Human Pluripotent Stem Cell Culture and Induced Pluripotent Stem Cell Derivation

    PubMed Central

    Vuoristo, Sanna; Toivonen, Sanna; Weltner, Jere; Mikkola, Milla; Ustinov, Jarkko; Trokovic, Ras; Palgi, Jaan; Lund, Riikka; Tuuri, Timo; Otonkoski, Timo

    2013-01-01

    Correct interactions with extracellular matrix are essential to human pluripotent stem cells (hPSC) to maintain their pluripotent self-renewal capacity during in vitro culture. hPSCs secrete laminin 511/521, one of the most important functional basement membrane components, and they can be maintained on human laminin 511 and 521 in defined culture conditions. However, large-scale production of purified or recombinant laminin 511 and 521 is difficult and expensive. Here we have tested whether a commonly available human choriocarcinoma cell line, JAR, which produces high quantities of laminins, supports the growth of undifferentiated hPSCs. We were able to maintain several human pluripotent stem cell lines on decellularized matrix produced by JAR cells using a defined culture medium. The JAR matrix also supported targeted differentiation of the cells into neuronal and hepatic directions. Importantly, we were able to derive new human induced pluripotent stem cell (hiPSC) lines on JAR matrix and show that adhesion of the early hiPSC colonies to JAR matrix is more efficient than to matrigel. In summary, JAR matrix provides a cost-effective and easy-to-prepare alternative for human pluripotent stem cell culture and differentiation. In addition, this matrix is ideal for the efficient generation of new hiPSC lines. PMID:24098444

  10. An Exploratory Investigation of the Relationships among Organizational Cultures, Organizational Communication Systems, and Member Satisfaction and Commitment.

    ERIC Educational Resources Information Center

    Pickett, Terry A.; Sorenson, Ritch L.

    A study examined the relationship between organizational communication systems and organizational culture and between culture and two human relations-based outcomes--satisfaction and commitment. Subjects were 26 college students who were assigned positions in one of two simulated, bureaucratic organizations that functioned for four class periods.…

  11. Joint-Service Integration: An Organizational Culture Study of the United States Department of Defense Voluntary Education System

    ERIC Educational Resources Information Center

    Benson, Martin K.

    2010-01-01

    The purpose of the descriptive case study with a multiple case framework was to (a) describe the organizational cultures of education programs and leaders in the United States (U.S.) Department of Defense (DoD) voluntary education system on Oahu, Hawaii; (b) determine if an overlapping common organizational culture exists; and (c) assess the…

  12. Comparing the Cultural Dimensions and Learners' Perceived Effectiveness of Online Learning Systems (OLS) among American and Malaysian Learners

    ERIC Educational Resources Information Center

    Keng, Seng C.

    2010-01-01

    With the rapid and exponential growth of Internet use worldwide, online learning has become one of the most widely used learning paradigms in the education environment. Yet despite the rapidly increasing cultural diversity of online learners, few studies have investigated the effectiveness of cross-cultural Online Learning Systems (OLS) using a…

  13. Analysis of cell identity, morphology, apoptosis and mitotic activity in a primary neural cell culture system in Drosophila

    PubMed Central

    2012-01-01

    In Drosophila, most neurogenetic research is carried out in vivo. Mammalian research demonstrates that primary cell culture techniques provide a powerful model to address cell autonomous and non-autonomous processes outside their endogenous environment. We developed a cell culture system in Drosophila using wildtype and genetically manipulated primary neural tissue for long-term observations. We assessed the molecular identity of distinct neural cell types by immunolabeling and genetically expressed fluorescent cell markers. We monitored mitotic activity of cell cultures derived from wildtype and tumorous larval brains. Our system provides a powerful approach to unveil developmental processes in the nervous system and to complement studies in vivo. PMID:22554060

  14. Tumor-suppressor effects of chemical functional groups in an in vitro co-culture system

    NASA Astrophysics Data System (ADS)

    Xu, Su-Ju; Cui, Fu-Zhai; Kong, Xiang-Dong

    2014-06-01

    Liver normal cells and cancer cells co-cultured on surfaces modified by different chemical functional groups, including mercapto (-SH), hydroxyl (-OH) and methyl (-CH3) groups. The results showed that different cells exhibited changes in response to different surfaces. Normal cells on -SH surface exhibited the smallest contact area with mostly rounded morphology, which led to the death of cancer cells, while cancer cells could not grow on -CH3 groups, which also died. In the co-culture system, the -CH3 group exhibited its unique effect that could trigger the death of cancer cells and had no effects on normal cells. Our findings provide useful information on strategies for the design of efficient and safe regenerative medicine materials.

  15. Seeing your way to health: the visual pedagogy of Bess Mensendieck's physical culture system.

    PubMed

    Veder, Robin

    2011-01-01

    This essay examines the images and looking practices central to Bess M. Mensendieck's (c.1866-1959) 'functional exercise' system, as documented in physical culture treatises published in Germany and the United States between 1906 and 1937. Believing that muscular realignment could not occur without seeing how the body worked, Mensendieck taught adult non-athletes to see skeletal alignment and muscular movement in their own and others' bodies. Three levels of looking practices are examined: didactic sequences; penetrating inspection and appreciation of physiological structures; and ideokinetic visual metaphors for guiding movement. With these techniques, Mensendieck's work bridged the body cultures of German Nacktkultur (nudism), American labour efficiency and the emerging physical education profession. This case study demonstrates how sport historians could expand their analyses to include practices of looking as well as questions of visual representation. PMID:21949947

  16. Pulse exposure of cultured rat neurons to aluminum-maltol affected the axonal transport system.

    PubMed

    Kashiwagi, Y; Nakamura, Y; Miyamae, Y; Hashimoto, R; Takeda, M

    1998-08-01

    Although chronic aluminum neurotoxicity has been well established, the mechanism of the toxicity has not been elucidated yet. In order to simplify the study of the aluminum neurotoxicity, we employed the pulse exposure of cultured rat cortical neurons to 250 microM aluminum-maltol for 1 h at the early stage (6 h after plating), which resulted in abnormal distribution of neurofilament L (NFL) and fast axonal transported proteins, whereas the axonal transport of tubulin, actin, and clathrin were not impaired. Otherwise, the pulse exposure of neurons at the late stage (4 days after plating) to the same concentration of aluminum-maltol did not affect the cell morphology and the distribution of NFL. The pulse exposure of cultured neurons to aluminum-maltol at the early stage might affect the axonal transport system of NFL and fast axonal transported proteins. PMID:9756345

  17. Genetically Encoded Sender–Receiver System in 3D Mammalian Cell Culture

    PubMed Central

    2013-01-01

    Engineering spatial patterning in mammalian cells, employing entirely genetically encoded components, requires solving several problems. These include how to code secreted activator or inhibitor molecules and how to send concentration-dependent signals to neighboring cells, to control gene expression. The Madin–Darby Canine Kidney (MDCK) cell line is a potential engineering scaffold as it forms hollow spheres (cysts) in 3D culture and tubulates in response to extracellular hepatocyte growth factor (HGF). We first aimed to graft a synthetic patterning system onto single developing MDCK cysts. We therefore developed a new localized transfection method to engineer distinct sender and receiver regions. A stable reporter line enabled reversible EGFP activation by HGF and modulation by a secreted repressor (a truncated HGF variant, NK4). By expanding the scale to wide fields of cysts, we generated morphogen diffusion gradients, controlling reporter gene expression. Together, these components provide a toolkit for engineering cell–cell communication networks in 3D cell culture. PMID:24313393

  18. Evaluation of mechanical and morphologic features of PLLA membranes as supports for perfusion cells culture systems.

    PubMed

    Montesanto, S; Brucato, V; La Carrubba, V

    2016-12-01

    Porous biodegradable PLLA membranes, which can be used as supports for perfusion cell culture systems were designed, developed and characterized. PLLA membranes were prepared via diffusion induced phase separation (DIPS). A glass slab was coated with a binary PLLA-dioxane solution (8wt.% PLLA) via dip coating, then pool immersed in two subsequent coagulation baths, and finally dried in a humidity-controlled environment. Surface and mechanical properties were evaluated by measuring pore size, porosity via scanning electron microscopy, storage modulus, loss modulus and loss angle by using a dynamic mechanical analysis (DMA). Cell adhesion assays on different membrane surfaces were also performed by using a standard count method. Results provide new insights into the foaming methods for producing polymeric membranes and supply indications on how to optimise the fabrication parameters to design membranes for tissue cultures and regeneration. PMID:27612778

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

    NASA Technical Reports Server (NTRS)

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

    2000-01-01

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

  20. The Comparative Evaluation of Antibacterial Activity of Methacryloxydodecyl Pyridinium Bromide and Non-methacryloxydodecyl Pyridinium Bromide Dentin Bonding Systems Using Two Different Techniques: An In vitro Study

    PubMed Central

    Deshpande, Prashanth; Nainan, Mohan Thomas; Metta, Kiran Kumar; Shivanna, Vasundhara; Ravi, Ramkrishna; Prashanth, B R

    2014-01-01

    Background: Adhesive systems have enabled clinicians to preserve more tooth structure by changing cavity designs. However, because of the polymerization shrinkage adhesive systems are not capable of totally prohibiting the gap formation between the cavity and restorative material of composite resin leading to colonization of oral microorganisms from saliva. One possible solution for this serious problem is to use dental materials with antibacterial properties. So the development of such agents has initiated for successful restorations. Hence, the purpose of this study was to compare the antibacterial activities of two dentin bonding systems: Clearfil protect bond (CPB) and prime & bond NT using agar well technique and tooth cavity model. Materials and Methods: CPB and prime and bond NT (PBNT) were evaluated in this study using agar well technique and tooth cavity model. In the agar well technique, the materials were filled in the wells of Muller-Hinton agar plates inoculated with Streptococcus mutans NCTC 10449M and the diameter of inhibition zones produced around the materials were measured after 24 h of incubation. For the tooth cavity model test, 3 cavities (of diameter - 1 mm and depth - 2 mm) were prepared in the flat occlusal dentin of human extracted molar. After sterilization, the teeth were left in the culture of broth of S. mutans at 37°C for 72 h for allowing bacteria to invade the cavity for 72 h. The dentin bonding systems were applied separately to each of the two infected cavities, and the third cavity was not applied and used as control. After sealing the occlusal surfaces, the teeth were kept in sterile physiological saline at 37°C for 72 h. The standardized amounts of dentin chips (120 + 5 mg) were obtained from the cavity walls, and the numbers of bacteria recovered were determined. The results were analyzed using one-way ANOVA, Kruskal–Wallis and Mann–Whitney’s U-tests. Results: The primer of CPB and PBNT produced similar inhibition

  1. Uncouplers of oxidative phosphorylation promote derepression of the hexose transport system in cultures of hamster cells.

    PubMed Central

    Kalckar, H M; Christopher, C W; Ullrey, D

    1979-01-01

    Long-term (18-24 hr) preincubation of NIL hamster cell cultures with D-glucose or D-glucosamine (both of which repress the hexose transport system) gave rise to a striking loss of the hexose transport system ("super-repression") when cycloheximide was also present in the culture medium. However, if 0.2 mM 2,4-dinitrophenol (DNP) was also present, the cycloheximide-mediated super-repression was prevented. Moreover, the presence of DNP at this low concentration contributed to an increase in hexose uptake such that it was substantially higher than that permitted by either of the two repressive sugars alone. When the cultures were maintained in medium containing D-fructose in place of glucose, a marked increase in uptake occurred, and this increase (derepression) was not affected by DNP. The derepression due to glucose deprivation and the increases caused by DNP treatment were also observed when 3-O-methylglucose was used to measure hexose transport. Although cultures maintained in the presence of glucosamine exhibited a repressed hexose transport rate, they did not generate significant amounts of lactic acid. DNP, and other uncouplers of oxidative phosphorylation, promoted a derepressed state of hexose transport but did not stimulate the generation of lactate from glucosamine. These data suggest that the metabolic repression phenomena of hexose transport do not depend on glycolysis but rather on the "energized" state of the cell. The energized state of the cell may also be required for the super-repression of hexose transport that is especially apparent when protein synthesis is blocked by cycloheximide. PMID:293732

  2. Development of an enhanced human gastrointestinal epithelial culture system to facilitate patient-based assays

    PubMed Central

    VanDussen, Kelli L; Marinshaw, Jeffrey M; Shaikh, Nurmohammad; Miyoshi, Hiroyuki; Moon, Clara; Tarr, Phillip I; Ciorba, Matthew A; Stappenbeck, Thaddeus S

    2014-01-01

    Objective The technology for the growth of human intestinal epithelial cells is rapidly progressing. An exciting possibility is that this system could serve as a platform for individualized medicine and research. However, to achieve this goal, human epithelial culture must be enhanced so that biopsies from individuals can be used to reproducibly generate cell lines in a short time frame so that multiple, functional assays can be performed (i.e., barrier function and host-microbial interactions). Design We created a large panel of human gastrointestinal epithelial cell lines (n = 65) from patient biopsies taken during routine upper and lower endoscopy procedures. Proliferative stem/progenitor cells were rapidly expanded using a high concentration of conditioned media containing the factors critical for growth (Wnt3a, R-spondin and Noggin). A combination of lower conditioned media concentration and Notch inhibition was used to differentiate these cells for additional assays. Results We obtained epithelial lines from all accessible tissue sites within two weeks of culture. The intestinal cell lines were enriched for stem cell markers and rapidly grew as spheroids that required passage at 1:3–1:4 every 3 days. Under differentiation conditions, intestinal epithelial spheroids showed region-specific development of mature epithelial lineages. These cells formed functional, polarized monolayers covered by a secreted mucus layer when grown on Transwell membranes. Using two-dimensional culture, these cells also demonstrated novel adherence phenotypes with various strains of pathogenic Escherichia coli. Conclusion This culture system will facilitate the study of inter-individual, functional studies of human intestinal epithelial cells, including host-microbial interactions. PMID:25007816

  3. Human whole-blood culture system for ex vivo characterization of designer-cell function.

    PubMed

    Schukur, Lina; Geering, Barbara; Fussenegger, Martin

    2016-03-01

    Encapsulated designer cells implanted into mice are currently used to validate the efficacy of therapeutic gene networks for the diagnosis and treatment of various human diseases in preclinical research. Because many human conditions cannot be adequately replicated by animal models, complementary and alternative procedures to test future treatment strategies are required. Here we describe a novel approach utilizing an ex vivo human whole-blood culture system to validate synthetic biology-inspired designer cell-based treatment strategies. The viability and functionality of transgenic mammalian designer cells co-cultured with primary human immune cells were characterized. We demonstrated that transgenic mammalian designer cells required adequate insulation from the human blood microenvironment to maintain viability and functionality. The biomaterial alginate-(poly-l-lysine)-alginate used to encapsulate the transgenic designer cells did neither affect the viability of primary granulocytes and lymphocytes nor the functionality of lymphocytes. Additionally, alginate-encapsulated transgenic designer cells remained responsive to the release of the pro-inflammatory cytokine tumor necrosis factor (TNF) from the whole-blood culture upon exposure to bacterial lipopolysaccharide (LPS). TNF diffused into the alginate capsules, bound to the specific TNF receptors on the transgenic designer cells' surface and triggered the expression of the reporter gene SEAP (human placental secreted alkaline phosphatase) that was rewired to the TNF-specific signaling cascade. Human whole-blood culture systems can therefore be considered as valuable complementary assays to animal models for the validation of synthetic circuits in genetically modified mammalian cells and may speed up preclinical research in a world of personalized medicine. PMID:26348251

  4. Comparison of Marine Microalgae Culture Systems for Fuels Production and Carbon Sequestration

    SciTech Connect

    Weissman, Joseph C; Polle, Juergen

    2006-05-30

    The dual problems of global fossil fuels supplies and global warming focus attention on the need to develop technologies that can provide large amounts of renewable fuels without contributing to global warming. The capture of power plant flue gas CO2 using microalgae cultures is one potential technology that could meet this objective. The central R&D issues are the design and operation of low-cost algal mass culture systems and the development of algal strains and cultivation techniques that can achieve very high biomass productivities. The major objective of this project was to develop mass culture techniques that could result in greatly increased biomass productivities, well above the about 50 metric tons per hectare per year (mt/ha/y) currently achievable. In this project, two marine microalgae species, the diatom Cyclotella sp.. and the green alga Tetraselmis sp., were cultivated on seawater in both open ponds and closed photo bioreactors, under a variety of different cultivation conditions. Simultaneous operation of the closed photo bioreactors and open ponds demonstrated similar productivities, under the same operating conditions. Thus the very expensive closed systems do not provide any major or inherent advantages in microalgae production over open ponds. Mutants of Cyclotella sp. were developed that exhibited reduced pigment content, which theoretically would result in greatly increased productivities when grown under full sunlight. However, in open ponds, these mutant strains exhibited similar productivities as the parental strains. The mutant strains all grew relatively slowly, suggesting that additional mutations masked whatever inherent potential for increased productivities may have resulted from the reduced pigment content. Research is still required to develop improved low pigment strains. When open pond cultures were exposed to intermittent sunlight, by partially covering the ponds with slats, solar conversion efficiencies increased dramatically

  5. Altered TNF-Alpha, Glucose, Insulin and Amino Acids in Islets Langerhans Cultured in a Microgravity Model System

    NASA Technical Reports Server (NTRS)

    Tobin, Brian W.; Leeper-Woodford, Sandra K.; Hashemi, Brian B.; Smith, Scott M.; Sams, Clarence F.

    2001-01-01

    The present studies were designed to determine effects of a microgravity model system upon lipopolysaccharide (LPS) stimulated tumor necrosis factor alpha (TNF-alpha) activity and indices of insulin and fuel homeostasis of pancreatic islets of Langerhans. Islets (1726+/-1 17,150 u IEU) from Wistar Furth rats were treated as: 1) HARV (High Aspect Ratio Vessel cell culture) , 2) HARV plus LPS, 3) static culture, 4) static culture plus LPS. TNF-alpha (L929 cytotoxicity assay) was significantly increased in LPS-induced HARV and static cultures, yet the increase was more pronounced in the static culture group (p<0.05). A decrease in insulin concentration was demonstrated in the LPS stimulated HARV culture (p<0.05). We observed a greater glucose concentration and increased disappearance of arginine in islets cultured in HARVs. While nitrogenous compound analysis indicated a ubiquitous reliance upon glutamine in all experimental groups, arginine was converted to ornithine at a two-fold greater rate in the islets cultured in the HARV microgravity model system (p<0.05). These studies demonstrate alterations in LPS induced TNF-alpha production of pancreatic islets of Langerhans, favoring a lesser TNF activity in the HARV. These alterations in fuel homeostasis may be promulgated by gravity averaged cell culture methods or by three dimensional cell assembly.

  6. A S.M.A.R.T. system for the seismic vulnerability mitigation of Cultural Heritages

    NASA Astrophysics Data System (ADS)

    Montuori, Antonio; Costanzo, Antonio; Gaudiosi, Iolanda; Vecchio, Antonio; Minasi, Mario; Falcone, Sergio; La Piana, Carmelo; Stramondo, Salvatore; Casula, Giuseppe; Giovanna Bianchi, Maria; Fabrizia Buongiorno, Maria; Musacchio, Massimo; Doumaz, Fawzi; Ilaria Pannaccione Apa, Maria

    2016-04-01

    Both assessment and mitigation of seismic vulnerability connected to cultural heritages monitoring are non-trivial issues, based on the knowledge of structural and environmental factors potential impacting the cultural heritage. A holistic approach could be suitable to provide an effective monitoring of cultural heritages within their surroundings at different spatial and temporal scales. On the one hand, the analysis about geometrical and structural properties of monuments is important to assess their state of conservation, their response to external stresses as well as anomalies related to natural and/or anthropogenic phenomena (e.g. the aging of materials, seismic stresses, vibrational modes). On the other hand, the investigation of the surrounding area is relevant to assess environmental properties and natural phenomena (e.g. landslides, earthquakes, subsidence, seismic response) as well as their related impacts on the monuments. Within such a framework, a multi-disciplinary system has been developed and here presented for the monitoring of cultural heritages for seismic vulnerability assessment and mitigation purposes*. It merges geophysical investigations and modeling, in situ measurements and multi-platforms remote sensing sensors for the non-destructive and non-invasive multi-scales monitoring of historic buildings in a seismic-prone area. In detail, the system provides: a) the long-term and the regional-scale analysis of buildings' environment through the integration of seismogenic analysis, airborne magnetic surveys, space-borne Synthetic Aperture Radar (SAR) and multi-spectral sensors. They allow describing the sub-surface fault systems, the surface deformation processes and the land use mapping of the regional-scale area on an annual temporal span; b) the short-term and the basin-scale analysis of building's neighborhood through geological setting and geotechnical surveys, airborne Light Detection And Radar (LiDAR) and ground-based SAR sensors. They

  7. Quantifying the Metrics That Characterize Safety Culture of Three Engineered Systems

    SciTech Connect

    Tucker, Julie; Ernesti, Mary; Tokuhiro, Akira

    2002-07-01

    With potential energy shortages and increasing electricity demand, the nuclear energy option is being reconsidered in the United States. Public opinion will have a considerable voice in policy decisions that will 'road-map' the future of nuclear energy in this country. This report is an extension of the last author's work on the 'safety culture' associated with three engineered systems (automobiles, commercial airplanes, and nuclear power plants) in Japan and the United States. Safety culture, in brief is defined as a specifically developed culture based on societal and individual interpretations of the balance of real, perceived, and imagined risks versus the benefits drawn from utilizing a given engineered systems. The method of analysis is a modified scale analysis, with two fundamental Eigen-metrics, time- (t) and number-scales (N) that describe both engineered systems and human factors. The scale analysis approach is appropriate because human perception of risk, perception of benefit and level of (technological) acceptance are inherently subjective, therefore 'fuzzy' and rarely quantifiable in exact magnitude. Perception of risk, expressed in terms of the psychometric factors 'dread risk' and 'unknown risk', contains both time- and number-scale elements. Various engineering system accidents with fatalities, reported by mass media are characterized by t and N, and are presented in this work using the scale analysis method. We contend that level of acceptance infers a perception of benefit at least two orders larger magnitude than perception of risk. The 'amplification' influence of mass media is also deduced as being 100- to 1000-fold the actual number of fatalities/serious injuries in a nuclear-related accident. (authors)

  8. Hetero-cellular prototyping by synchronized multi-material bioprinting for rotary cell culture system.

    PubMed

    Snyder, Jessica; Son, Ae Rin; Hamid, Qudus; Wu, Honglu; Sun, Wei

    2016-03-01

    Bottom-up tissue engineering requires methodological progress of biofabrication to capture key design facets of anatomical arrangements across micro, meso and macro-scales. The diffusive mass transfer properties necessary to elicit stability and functionality require hetero-typic contact, cell-to-cell signaling and uniform nutrient diffusion. Bioprinting techniques successfully build mathematically defined porous architecture to diminish resistance to mass transfer. Current limitations of bioprinted cell assemblies include poor micro-scale formability of cell-laden soft gels and asymmetrical macro-scale diffusion through 3D volumes. The objective of this work is to engineer a synchronized multi-material bioprinter (SMMB) system which improves the resolution and expands the capability of existing bioprinting systems by packaging multiple cell types in heterotypic arrays prior to deposition. This unit cell approach to arranging multiple cell-laden solutions is integrated with a motion system to print heterogeneous filaments as tissue engineered scaffolds and nanoliter droplets. The set of SMMB process parameters control the geometric arrangement of the combined flow's internal features and constituent material's volume fractions. SMMB printed hepatocyte-endothelial laden 200 nl droplets are cultured in a rotary cell culture system (RCCS) to study the effect of microgravity on an in vitro model of the human hepatic lobule. RCCS conditioning for 48 h increased hepatocyte cytoplasm diameter 2 μm, increased metabolic rate, and decreased drug half-life. SMMB hetero-cellular models present a 10-fold increase in metabolic rate, compared to SMMB mono-culture models. Improved bioprinting resolution due to process control of cell-laden matrix packaging as well as nanoliter droplet printing capability identify SMMB as a viable technique to improve in vitro model efficacy. PMID:26759993

  9. A Method for Ovarian Follicle Encapsulation and Culture in a Proteolytically Degradable 3 Dimensional System

    PubMed Central

    Shikanov, Ariella; Xu, Min; Woodruff, Teresa K.; Shea, Lonnie D.

    2011-01-01

    The ovarian follicle is the functional unit of the ovary that secretes sex hormones and supports oocyte maturation. In vitro follicle techniques provide a tool to model follicle development in order to investigate basic biology, and are further being developed as a technique to preserve fertility in the clinic1-4. Our in vitro culture system employs hydrogels in order to mimic the native ovarian environment by maintaining the 3D follicular architecture, cell-cell interactions and paracrine signaling that direct follicle development 5. Previously, follicles were successfully cultured in alginate, an inert algae-derived polysaccharide that undergoes gelation with calcium ions6-8. Alginate hydrogels formed at a concentration of 0.25% w/v were the most permissive for follicle culture, and retained the highest developmental competence 9. Alginate hydrogels are not degradable, thus an increase in the follicle diameter results in a compressive force on the follicle that can impact follicle growth10. We subsequently developed a culture system based on a fibrin-alginate interpenetrating network (FA-IPN), in which a mixture of fibrin and alginate are gelled simultaneously. This combination provides a dynamic mechanical environment because both components contribute to matrix rigidity initially; however, proteases secreted by the growing follicle degrade fibrin in the matrix leaving only alginate to provide support. With the IPN, the alginate content can be reduced below 0.25%, which is not possible with alginate alone 5. Thus, as the follicle expands, it will experience a reduced compressive force due to the reduced solids content. Herein, we describe an encapsulation method and an in vitro culture system for ovarian follicles within a FA-IPN. The dynamic mechanical environment mimics the natural ovarian environment in which small follicles reside in a rigid cortex and move to a more permissive medulla as they increase in size11. The degradable component may be particularly

  10. Another Route to a General Theory of Cultural Transmission: A Systems Model

    ERIC Educational Resources Information Center

    Dobbert, Marion L.

    1975-01-01

    A cultural transmission theory is presented. The model has two uses: to provide a guide for gathering cultural transmission data; and to set up a computerized cultural transmission simulation. Availability information is given in SO 504 073. (Author/RM)

  11. Effect of different culture systems on the production of foot and mouth disease trivalent vaccine

    PubMed Central

    Hassan, Amr Ismail

    2016-01-01

    Aim: This study aims to determine the effect of the stationary rawx, roller, and the suspension cell culture systems on the total virus yield infectivity and antigenicity. Materials and Methods: Three serotypes of foot and mouth disease virus (FMDV) (serotype A, O and SAT-2) were inoculated separately into baby hamster kidney-21 cell line in rawx, roller, and suspension cultivation systems using multiplicity of infection (1:100). Samples were taken from the total virus yield from each system at 15, 18, 21, and 24 h post-inoculation. Testing the total virus yield infectivity through virus titration and antigenicity through estimation of complement fixing titer and 146S content and evaluation of the potency of the vaccine prepared from the different cultivation systems were done. Results: The results showed that the FMDV titer of serotype A, O, and SAT-2 obtained from the roller cultivation system showed the highest level followed by suspension cultivation system then the rawx cultivation system. The FMDV titer showed its highest level at 21 h post-inoculation in all the cultivation systems and then decline at 24 h post-inoculation. The antigenicity reached its highest value content at 18 h post-inoculation either by complement fixation test or by quantifying the 146S intact virion. Montanide ISA 206 oil inactivated trivalent vaccines were prepared from the tested serotypes (A Iran O5. O Panasia and SAT-2/EGY/2012) harvested at 18 h post-inoculation from the 3 culture systems. The results of tracing the antibody response showed that the mean antibody response from the roller cultivation system start its protective antibody titer earlier at 2 weeks post-vaccination (WPV) than the vaccine prepared from the other two cultivation system and the immune protection period lasts longer for 36 WPV for the roller cultivation system vaccine than the other two cultivation systems. Conclusion: The best cultivation system used for the production of FMD vaccine regarding its

  12. A Scalable Perfusion Culture System with Miniature Peristaltic Pumps for Live-Cell Imaging Assays with Provision for Microfabricated Scaffolds

    PubMed Central

    Balakrishnan, Sreenath; Suma, M.S.; Raju, Shilpa R.; Bhargav, Santosh D.B.; Arunima, S.; Das, Saumitra

    2015-01-01

    Abstract We present a perfusion culture system with miniature bioreactors and peristaltic pumps. The bioreactors are designed for perfusion, live-cell imaging studies, easy incorporation of microfabricated scaffolds, and convenience of operation in standard cell culture techniques. By combining with miniature peristaltic pumps—one for each bioreactor to avoid cross-contamination and to maintain desired flow rate in each—we have made a culture system that facilitates perfusion culture inside standard incubators. This scalable system can support multiple parallel perfusion experiments. The major components are fabricated by three-dimensional printing using VeroWhite, which we show to be amenable to ex vivo cell culture. Furthermore, the components of the system can be reused, thus making it economical. We validate the system and illustrate its versatility by culturing primary rat hepatocytes, live imaging the growth of mouse fibroblasts (NIH 3T3) on microfabricated ring-scaffolds inserted into the bioreactor, performing perfusion culture of breast cancer cells (MCF7), and high-magnification imaging of hepatocarcinoma cells (HuH7). PMID:26309810

  13. Growth of sweetpotato cultured in the newly designed hydroponic system for space farming

    NASA Astrophysics Data System (ADS)

    Kitaya, Y.; Hirai, H.; Wei, X.; Islam, A. F. M. S.; Yamamoto, M.

    Life support of crews in long-duration space missions for other planets will be highly dependent on amounts of food, atmospheric O2 and clean water produced by plants. Therefore, the space farming system with scheduling of crop production, obtaining high yields with a rapid turnover rate, converting atmospheric CO2 to O2 and purifying water should be established with employing suitable plant species and cultivars and precisely controlling environmental variables around plants grown at a high density in a limited space. In this study, we developed a new hydroponic method for producing tuberous roots and fresh edible leaves and stems of sweetpotato. In the first experiment, we examined the effects of water contents in the rooting substrate on growth and tuberous root development of sweetpotato. The rooting substrates made with rockwool slabs were inclined in a culture container and absorbed nutrient solution from the lower end of the slabs by capillary action. Tuberous roots developed on the lower surface of the rockwool slabs. The tuberous roots showed better growth and development at locations farther from the water surface on the rockwool slabs, which had lower water content. In the second experiment, three sweetpotato cultivars were cultured in a hydroponic system for five months from June to November under the sun light in Osaka, Japan as a fundamental study for establishing the space farming system. The cultivars employed were ‘Elegant summer’, ‘Kokei-14’ and ‘Beniazuma’. The hydroponic system mainly consisted of culture containers and rockwool slabs. Dry weights of tuberous roots developed in the aerial space between the rockwool slab and the nutrient solution filled at the bottom of the culture container were 0.34, 0.45 and 0.23 kg/plant and dry weights of the top portion (leaves, petioles and stems) were 0.42, 0.29 and 0.61 kg/plant for ‘Elegant summer’, ‘Kokei-14’ and ‘Beniazuma’, respectively. Young stems and leaves as well as

  14. Adipose Tissue Engineering in Three-Dimensional Levitation Tissue Culture System Based on Magnetic Nanoparticles

    PubMed Central

    Daquinag, Alexes C.; Souza, Glauco R.

    2013-01-01

    White adipose tissue (WAT) is becoming widely used in regenerative medicine/cell therapy applications, and its physiological and pathological importance is increasingly appreciated. WAT is a complex organ composed of differentiated adipocytes, stromal mesenchymal progenitors known as adipose stromal cells (ASC), as well as endothelial vascular cells and infiltrating leukocytes. Two-dimensional (2D) culture that has been typically used for studying adipose cells does not adequately recapitulate WAT complexity. Improved methods for reconstruction of functional WAT ex vivo are instrumental for understanding of physiological interactions between the composing cell populations. Here, we used a three-dimensional (3D) levitation tissue culture system based on magnetic nanoparticle assembly to model WAT development and growth in organoids termed adipospheres. We show that 3T3-L1 preadipocytes remain viable in spheroids for a long period of time, while in 2D culture, they lose adherence and die after reaching confluence. Upon adipogenesis induction in 3T3-L1 adipospheres, cells efficiently formed large lipid droplets typical of white adipocytes in vivo, while only smaller lipid droplet formation is achievable in 2D. Adiposphere-based coculture of 3T3-L1 preadipocytes with murine endothelial bEND.3 cells led to a vascular-like network assembly concomitantly with lipogenesis in perivascular cells. Adipocyte-depleted stromal vascular fraction (SVF) of mouse WAT cultured in 3D underwent assembly into organoids with vascular-like structures containing luminal endothelial and perivascular stromal cell layers. Adipospheres made from primary WAT cells displayed robust proliferation and complex hierarchical organization reflected by a matricellular gradient incorporating ASC, endothelial cells, and leukocytes, while ASC quickly outgrew other cell types in adherent culture. Upon adipogenesis induction, adipospheres derived from the SVF displayed more efficient lipid droplet

  15. Voluntary organ donation system adapted to Chinese cultural values and social reality.

    PubMed

    Huang, Jiefu; Millis, J Michael; Mao, Yilei; Millis, M Andrew; Sang, Xinting; Zhong, Shouxian

    2015-04-01

    Organ donation and transplant systems have unique characteristics based on the local culture and socioeconomic context. China's transplant and organ donation systems developed without regulatory oversight until 2006 when regulation and policy were developed and then implemented over the next several years. Most recently, the pilot project of establishing a voluntary citizen-based deceased donor program was established. The pilot program addressed the legal, financial, and cultural barriers to organ donation in China. The pilot program has evolved into a national program. Significantly, it established a uniquely Chinese donor classification system. The Chinese donor classification system recognizes donation after brain death (category I), donation after circulatory death (category II), and donation after brain death followed by circulatory death (category III). Through August 2014, the system has identified 2326 donors and provided 6416 organs that have been allocated though a transparent organ allocation system. The estimated number of donors in 2014 is 1147. As China's attitudes toward organ donation have matured and evolved and as China, as a nation, is taking its place on the world stage, it is recognizing that its past practice of using organs from executed prisoners is not sustainable. It is time to recognize that the efforts to regulate transplantation and provide voluntary citizen-based deceased organ donation have been successful and that China should use this system to provide organs for all transplants in every province and hospital in China. At the national organ transplant congress on October 30, 2014, the Chairman of the China's national organ donation and transplantation committee, Jeifu Huang required all hospitals to stop using organs from executed prisoners immediately and the civilian organ donation will be sole source for organ transplant in China starting January 2015. PMID:25545626

  16. A Continuous Culture System for Assessing Microbial Activities in the Piezosphere

    PubMed Central

    Pérez-Rodríguez, Ileana

    2015-01-01

    Continuous culture under elevated pressures is an important technique for expanding the exploration of microbial growth and survival in extreme environments associated with the deep biosphere. Here we present a benchtop stirred continuous culture bioreactor capable of withstanding temperatures ranging from 25 to 120°C and pressures as high as 69 MPa. The system is configured to allow the employment of media enriched in dissolved gases, under oxic or anoxic conditions, while permitting periodic sampling of the incubated organisms with minimal physical/chemical disturbance inside the reactor. In a pilot experiment, the fermentative growth of the thermopiezophilic bacterium Marinitoga piezophila was investigated continuously for 382 h at 65°C and at pressures ranging from 0.1 to 40 MPa while the medium flow rate was varied from 2 to 0.025 ml/min. The enhanced growth observed at 30 and 40 MPa and 0.025 ml/min supports the pressure preferences of M. piezophila when grown fermentatively. This assay successfully demonstrates the capabilities of the bioreactor for continuous culturing at a variety of dilution rates, pressures, and temperatures. We anticipate that this technology will accelerate our understanding of the physiological and metabolic status of microorganisms under temperature, pressure, and energy regimes resembling those of the Earth's piezosphere. PMID:26209666

  17. A Continuous Culture System for Assessing Microbial Activities in the Piezosphere.

    PubMed

    Foustoukos, Dionysis I; Pérez-Rodríguez, Ileana

    2015-10-01

    Continuous culture under elevated pressures is an important technique for expanding the exploration of microbial growth and survival in extreme environments associated with the deep biosphere. Here we present a benchtop stirred continuous culture bioreactor capable of withstanding temperatures ranging from 25 to 120°C and pressures as high as 69 MPa. The system is configured to allow the employment of media enriched in dissolved gases, under oxic or anoxic conditions, while permitting periodic sampling of the incubated organisms with minimal physical/chemical disturbance inside the reactor. In a pilot experiment, the fermentative growth of the thermopiezophilic bacterium Marinitoga piezophila was investigated continuously for 382 h at 65°C and at pressures ranging from 0.1 to 40 MPa while the medium flow rate was varied from 2 to 0.025 ml/min. The enhanced growth observed at 30 and 40 MPa and 0.025 ml/min supports the pressure preferences of M. piezophila when grown fermentatively. This assay successfully demonstrates the capabilities of the bioreactor for continuous culturing at a variety of dilution rates, pressures, and temperatures. We anticipate that this technology will accelerate our understanding of the physiological and metabolic status of microorganisms under temperature, pressure, and energy regimes resembling those of the Earth's piezosphere. PMID:26209666

  18. Studies on luminous, Vibrio harveyi associated with shrimp culture system rearing Penaeus monodon.

    PubMed

    Kannapiran, E; Ravindran, J; Chandrasekar, R; Kalaiarasi, A

    2009-09-01

    Microbiological studies in a modified extensive shrimp culture system at Nambuthalai, southeast coast of India were carried out fora period of 120 days. Population dynamics and distribution profile of luminous bacteria and total heterotrophic bacteria in the water sediment and animal samples were monitored. Luminous bacteria associated with exoskeleton, gills and gut were isolated and quantified. The total heterotrophic bacterial counts ranged from 1.3 x 10(4) to 25.3 x 10(4) CFU ml(-1) in water and 1.5 x 10(6) to 26.2 x 10(6) CFU g(-1) in sediment. The V. harveyi population density varied between 0.6 x 10(4) and 8.8 x 10(4) LCFU ml(-1) in water and from 1.2 x 10(6) to 10.4 x 10(6) LCFU g(-1) sediment respectively. The gut of the animal was found to harbor high density of V. harveyi than gills and exoskeleton. The total heterotrophic bacteria and V. harveyi population density showed increasing trend during the culture period. The high V harveyi density observed in this study at the end of the culture period correlated with the outbreak of white spot disease. PMID:20143707

  19. Columbia River System Operation Review : Final Environmental Impact Statement, Appendix D: Cultural Resources.

    SciTech Connect

    Columbia River System Operation Review

    1995-11-01

    This study attempts to identify and analyze the impacts of the System Operating Strategy (SOS) alternatives on cultural resources. The impacts include effects on Native American traditional cultural values, properties and practices. They also include effects on archeological or historic properties meeting the criteria of the National Register of Historic Places. In addition to responding to the requirements of the National Environmental Policy Act (NEPA), this analysis addresses the requirements of the National Historic Preservation Act (NHPA), the Archeological Resources Protection Act (ARPA), the Native American Graves Protection and Repatriation Act (NAGPRA), the Native American Religious Freedom Act (NARFA), and other relevant legislation. To meet their legally mandated cultural resources requirements, the SOR agencies will develop agreements and Implementation Plans with the appropriate State Historic Preservation Officers (SHPOs), Tribes, and the Advisory Council on Historic Preservation (ACHP) detailing the measures necessary to best manage the resource. The planning and implementation activities will be staged over a number of years in consultation with affected Tribes.

  20. A 3D human neural cell culture system for modeling Alzheimer’s disease

    PubMed Central

    Kim, Young Hye; Choi, Se Hoon; D’Avanzo, Carla; Hebisch, Matthias; Sliwinski, Christopher; Bylykbashi, Enjana; Washicosky, Kevin J.; Klee, Justin B.; Brüstle, Oliver; Tanzi, Rudolph E.; Kim, Doo Yeon

    2015-01-01

    Stem cell technologies have facilitated the development of human cellular disease models that can be used to study pathogenesis and test therapeutic candidates. These models hold promise for complex neurological diseases such as Alzheimer’s disease (AD) because existing animal models have been unable to fully recapitulate all aspects of pathology. We recently reported the characterization of a novel three-dimensional (3D) culture system that exhibits key events in AD pathogenesis, including extracellular aggregation of β-amyloid and accumulation of hyperphosphorylated tau. Here we provide instructions for the generation and analysis of 3D human neural cell cultures, including the production of genetically modified human neural progenitor cells (hNPCs) with familial AD mutations, the differentiation of the hNPCs in a 3D matrix, and the analysis of AD pathogenesis. The 3D culture generation takes 1–2 days. The aggregation of β-amyloid is observed after 6-weeks of differentiation followed by robust tau pathology after 10–14 weeks. PMID:26068894

  1. Revisiting the Diego Blood Group System in Amerindians: Evidence for Gene-Culture Comigration.

    PubMed

    Bégat, Christophe; Bailly, Pascal; Chiaroni, Jacques; Mazières, Stéphane

    2015-01-01

    Six decades ago the DI*A allele of the Diego blood group system was instrumental in proving Native American populations originated from Siberia. Since then, it has received scant attention. The present study was undertaken to reappraise distribution of the DI*A allele in 144 Native American populations based on current knowledge. Using analysis of variance tests, frequency distribution was studied according to geographical, environmental, and cultural parameters. Frequencies were highest in Amazonian populations. In contrast, DI*A was undetectable in subarctic, Fuegian, Panamanian, Chaco and Yanomama populations. Closer study revealed a correlation that this unequal distribution was correlated with language, suggesting that linguistic divergence was a driving force in the expansion of DI*A among Native Americans. The absence of DI*A in circumpolar Eskimo-Aleut and Na-Dene speakers was consistent with a late migratory event confined to North America. Distribution of DI*A in subtropical areas indicated that gene and culture exchanges were more intense within than between ecozones. Bolstering the utility of classical genetic markers in biological anthropology, the present study of the expansion of Diego blood group genetic polymorphism in Native Americans shows strong evidence of gene-culture comigration. PMID:26148209

  2. A 3D human neural cell culture system for modeling Alzheimer's disease.

    PubMed

    Kim, Young Hye; Choi, Se Hoon; D'Avanzo, Carla; Hebisch, Matthias; Sliwinski, Christopher; Bylykbashi, Enjana; Washicosky, Kevin J; Klee, Justin B; Brüstle, Oliver; Tanzi, Rudolph E; Kim, Doo Yeon

    2015-07-01

    Stem cell technologies have facilitated the development of human cellular disease models that can be used to study pathogenesis and test therapeutic candidates. These models hold promise for complex neurological diseases such as Alzheimer's disease (AD), because existing animal models have been unable to fully recapitulate all aspects of pathology. We recently reported the characterization of a novel 3D culture system that exhibits key events in AD pathogenesis, including extracellular aggregation of amyloid-β (Aβ) and accumulation of hyperphosphorylated tau. Here we provide instructions for the generation and analysis of 3D human neural cell cultures, including the production of genetically modified human neural progenitor cells (hNPCs) with familial AD mutations, the differentiation of the hNPCs in a 3D matrix and the analysis of AD pathogenesis. The 3D culture generation takes 1-2 d. The aggregation of Aβ is observed after 6 weeks of differentiation, followed by robust tau pathology after 10-14 weeks. PMID:26068894

  3. Glycolysis inhibition by palmitate in renal cells cultured in a two-chamber system.

    PubMed

    Bolon, C; Gauthier, C; Simonnet, H

    1997-11-01

    A major shortcoming of renal proximal tubular cells (RPTC) in culture is the gradual modification of their energy metabolism from the oxidative type to the glycolytic type. To test the possible reduction of glycolysis by naturally occurring long-chain fatty acids, RPTC were cultured in a two-chamber system, with albumin-bound palmitate (0.4 mM) added to the basolateral chamber after confluency. Twenty-four hours of contact with palmitate decreased glycolysis by 38% provided that carnitine was present; lactate production was decreased by 38%, and the decrease in glycolysis resulted from a similar decrease of basolateral and apical net uptake of glucose. In contrast to the previously described effect of the nonphysiological oxidative substrate heptanoate, palmitate promoted a long-term decrease in lactate production and sustained excellent cellular growth. After 4 days of contact, decreased glycolysis was maintained even in the absence of carnitine and resulted from a decrease of basolateral uptake only, suggestive of long-term regulation different from the earlier effects. Thus, although cultured RPTC lost their oxidative phenotype, they exhibited a type of regulation (Randle effect) that is found in the oxidative-type but not in the glycolytic-type tissues, therefore unmasking a regulative capacity barely detectable in fresh RPTC. Low PO2 (50 mmHg in the apical chamber) could be a major cause of elevated glycolysis and could hinder the effects of palmitate. PMID:9374661

  4. Revisiting the Diego Blood Group System in Amerindians: Evidence for Gene-Culture Comigration

    PubMed Central

    Bégat, Christophe; Bailly, Pascal; Chiaroni, Jacques; Mazières, Stéphane

    2015-01-01

    Six decades ago the DI*A allele of the Diego blood group system was instrumental in proving Native American populations originated from Siberia. Since then, it has received scant attention. The present study was undertaken to reappraise distribution of the DI*A allele in 144 Native American populations based on current knowledge. Using analysis of variance tests, frequency distribution was studied according to geographical, environmental, and cultural parameters. Frequencies were highest in Amazonian populations. In contrast, DI*A was undetectable in subarctic, Fuegian, Panamanian, Chaco and Yanomama populations. Closer study revealed a correlation that this unequal distribution was correlated with language, suggesting that linguistic divergence was a driving force in the expansion of DI*A among Native Americans. The absence of DI*A in circumpolar Eskimo-Aleut and Na-Dene speakers was consistent with a late migratory event confined to North America. Distribution of DI*A in subtropical areas indicated that gene and culture exchanges were more intense within than between ecozones. Bolstering the utility of classical genetic markers in biological anthropology, the present study of the expansion of Diego blood group genetic polymorphism in Native Americans shows strong evidence of gene-culture comigration. PMID:26148209

  5. 3-Dimensional culture systems for anti-cancer compound profiling and high-throughput screening reveal increases in EGFR inhibitor-mediated cytotoxicity compared to monolayer culture systems.

    PubMed

    Howes, Amy L; Richardson, Robyn D; Finlay, Darren; Vuori, Kristiina

    2014-01-01

    3-dimensional (3D) culture models have the potential to bridge the gap between monolayer cell culture and in vivo studies. To benefit anti-cancer drug discovery from 3D models, new techniques are needed that enable their use in high-throughput (HT) screening amenable formats. We have established miniaturized 3D culture methods robust enough for automated HT screens. We have applied these methods to evaluate the sensitivity of normal and tumorigenic breast epithelial cell lines against a panel of oncology drugs when cultured as monolayers (2D) and spheroids (3D). We have identified two classes of compounds that exhibit preferential cytotoxicity against cancer cells over normal cells when cultured as 3D spheroids: microtubule-targeting agents and epidermal growth factor receptor (EGFR) inhibitors. Further improving upon our 3D model, superior differentiation of EC50 values in the proof-of-concept screens was obtained by co-culturing the breast cancer cells with normal human fibroblasts and endothelial cells. Further, the selective sensitivity of the cancer cells towards chemotherapeutics was observed in 3D co-culture conditions, rather than as 2D co-culture monolayers, highlighting the importance of 3D cultures. Finally, we examined the putative mechanisms that drive the differing potency displayed by EGFR inhibitors. In summary, our studies establish robust 3D culture models of human cells for HT assessment of tumor cell-selective agents. This methodology is anticipated to provide a useful tool for the study of biological differences within 2D and 3D culture conditions in HT format, and an important platform for novel anti-cancer drug discovery. PMID:25247711

  6. Impact of depuration of earthy-musty off-flavors on fillet quality of Atlantic salmon, Salmo salar, cultured in a recirculating system

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Over the past decade in the United States, there has been increased interest in the establishment and use of land-based, closed-containment systems [e.g., recirculating aquaculture system (RAS)] for salmonid culture. These culture systems have unique challenges compared to net pen culture of salmon...

  7. Impact of depuration of earth-musty off-flavors on fillet quality of atlantic salmon, Salmo salar, cultured in a recirculating aquaculture system

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Over the past decade in the United States, there has been increased interest in the establishment and use of land-based, closed-containment systems [e.g., recirculating aquaculture system (RAS)] for salmonid culture. These culture systems have unique challenges compared to net pen culture of salmon...

  8. Multilineage differentiation of rhesus monkey embryonic stem cells in three-dimensional culture systems

    NASA Technical Reports Server (NTRS)

    Chen, Silvia S.; Revoltella, Roberto P.; Papini, Sandra; Michelini, Monica; Fitzgerald, Wendy; Zimmerberg, Joshua; Margolis, Leonid

    2003-01-01

    In the course of normal embryogenesis, embryonic stem (ES) cells differentiate along different lineages in the context of complex three-dimensional (3D) tissue structures. In order to study this phenomenon in vitro under controlled conditions, 3D culture systems are necessary. Here, we studied in vitro differentiation of rhesus monkey ES cells in 3D collagen matrixes (collagen gels and porous collagen sponges). Differentiation of ES cells in these 3D systems was different from that in monolayers. ES cells differentiated in collagen matrixes into neural, epithelial, and endothelial lineages. The abilities of ES cells to form various structures in two chemically similar but topologically different matrixes were different. In particular, in collagen gels ES cells formed gland-like circular structures, whereas in collagen sponges ES cells were scattered through the matrix or formed aggregates. Soluble factors produced by feeder cells or added to the culture medium facilitated ES cell differentiation into particular lineages. Coculture with fibroblasts in collagen gel facilitated ES cell differentiation into cells of a neural lineage expressing nestin, neural cell adhesion molecule, and class III beta-tubulin. In collagen sponges, keratinocytes facilitated ES cell differentiation into cells of an endothelial lineage expressing factor VIII. Exogenous granulocyte-macrophage colony-stimulating factor further enhanced endothelial differentiation. Thus, both soluble factors and the type of extracellular matrix seem to be critical in directing differentiation of ES cells and the formation of tissue-like structures. Three-dimensional culture systems are a valuable tool for studying the mechanisms of these phenomena.

  9. Implementing oxygen control in chip-based cell and tissue culture systems.

    PubMed

    Oomen, Pieter E; Skolimowski, Maciej D; Verpoorte, Elisabeth

    2016-09-21

    Oxygen is essential in the energy metabolism of cells, as well as being an important regulatory parameter influencing cell differentiation and function. Interest in precise oxygen control for in vitro cultures of tissues and cells continues to grow, especially with the emergence of the organ-on-a-chip and the desire to emulate in vivo conditions. This was recently discussed in this journal in a Critical Review by Brennan et al. (Lab Chip (2014). DOI: ). Microfluidics can be used to introduce flow to facilitate nutrient supply to and waste removal from in vitro culture systems. Well-defined oxygen gradients can also be established. However, cells can quickly alter the oxygen balance in their vicinity. In this Tutorial Review, we expand on the Brennan paper to focus on the implementation of oxygen analysis in these systems to achieve continuous monitoring. Both electrochemical and optical approaches for the integration of oxygen monitoring in microfluidic tissue and cell culture systems will be discussed. Differences in oxygen requirements from one organ to the next are a challenging problem, as oxygen delivery is limited by its uptake into medium. Hence, we discuss the factors determining oxygen concentrations in solutions and consider the possible use of artificial oxygen carriers to increase dissolved oxygen concentrations. The selection of device material for applications requiring precise oxygen control is discussed in detail, focusing on oxygen permeability. Lastly, a variety of devices is presented, showing the diversity of approaches that can be employed to control and monitor oxygen concentrations in in vitro experiments. PMID:27492338

  10. Bridging the gap between traditional cell cultures and bioreactors applied in regenerative medicine: practical experiences with the MINUSHEET perfusion culture system.

    PubMed

    Minuth, Will W; Denk, Lucia

    2016-03-01

    To meet specific requirements of developing tissues urgently needed in tissue engineering, biomaterial research and drug toxicity testing, a versatile perfusion culture system was developed. First an individual biomaterial is selected and then mounted in a MINUSHEET(®) tissue carrier. After sterilization the assembly is transferred by fine forceps to a 24 well culture plate for seeding cells or mounting tissue on it. To support spatial (3D) development a carrier can be placed in various types of perfusion culture containers. In the basic version a constant flow of culture medium provides contained tissue with always fresh nutrition and respiratory gas. For example, epithelia can be transferred to a gradient container, where they are exposed to different fluids at the luminal and basal side. To observe development of tissue under the microscope, in a different type of container a transparent lid and base are integrated. Finally, stem/progenitor cells are incubated in a container filled by an artificial interstitium to support spatial development. In the past years the described system was applied in numerous own and external investigations. To present an actual overview of resulting experimental data, the present paper was written. PMID:25894791

  11. Denitrification with epsilon-caprolactam by acclimated mixed culture and by pure culture of bacteria isolated from polyacrylonitrile fibre manufactured wastewater treatment system.

    PubMed

    Lee, C M; Wang, C C

    2004-01-01

    The aim of this study is to isolate denitrifying bacteria utilizing epsilon-caprolactam as the substrate, from a polyacrylonitrile fibre manufactured wastewater treatment system. The aim is also to compare the performance of PAN (polyacrylonitrile) mixed bacteria cultures acclimated to epsilon-caprolactam and isolated pure strain for treating different initial epsilon-caprolactam concentrations from synthetic wastewater under anoxic conditions. The result showed that the PAN mixed bacteria cultures acclimated to epsilon-caprolactam could utilize 1538.5 mg/l of epsilon-caprolactam as a substrate for denitrification. Sufficient time and about 2200 mg/l of nitrate were necessary for the complete epsilon-caprolactam removal. Paracoccus thiophilus was isolated from the polyacrylonitrile fibre manufactured wastewater treatment system and it could utilize 1722.5 mg/l of epsilon-caprolactam as a substrate for denitrification. About 3500 mg/l of nitrate was necessary for the complete removal of epsilon-caprolactam. When the initial epsilon-caprolactam concentration was below 784.3 mg/l, the removal efficiency of epsilon-caprolactam by Paracoccus thiophilus was better than that for the PAN mixed bacteria cultures. The growth of Paracoccus thiophilus was better. However, when the initial epsilon-caprolactam concentration was as high as 1445.8 mg/l, both the epsilon-caprolactam removal efficiency by Paracoccus thiophilus and Paracoccus thiophilus specific growth rate were similar to the PAN mixed bacteria cultures. PMID:15137443

  12. Are the core values of the radiological protection system shared across cultures?

    PubMed

    Zölzer, F

    2016-06-01

    In spite of ongoing globalisation in many fields, the ethics of radiological protection have long been discussed almost exclusively in terms of 'Western' moral philosophy concepts such as utilitarianism or deontology. A cross-cultural discourse in this field is only just beginning. In 'Principles of Biomedical Ethics', Beauchamp and Childress suggested that there exists a 'common morality' which is 'not relative to cultures or individuals, because it transcends both'. They proposed four cross-culturally valid principles for decision making in medicine: respect for autonomy, non-maleficence, beneficence, and justice. A similar approach is being developed by the International Commission on Radiological Protection Task Group 94 on the ethics of radiological protection. Here, the core values are: human dignity, beneficence/non-maleficence, prudence, and justice. Other values could be added, such as consideration for the interests of society as a whole or the interests of future generations, or procedural values such as transparency and accountability; this paper will include a brief discussion on how they relate to the four basic principles. The main question to be addressed here, however, is whether the proposed core values are indeed part of a 'common morality'. This, as it will be argued, cannot be decided by a global opinion poll, but has to be based on an analysis of the written and oral traditions that have provided ethical orientation throughout history, and are still considered seminal by the majority of people. It turns out that there are indeed many commonalities across cultures, and that the concept of globally shared core values for the radiological protection system is not hopelessly idealistic. PMID:26984903

  13. Generation of Sheffield (Shef) human embryonic stem cell lines using a microdrop culture system.

    PubMed

    Aflatoonian, Behrouz; Ruban, Ludmila; Shamsuddin, Shamsul; Baker, Duncan; Andrews, Peter; Moore, Harry

    2010-04-01

    The conventional method for the derivation of human embryonic stem cells (hESCs) involves inner cell mass (ICM) co-culture with a feeder layer of inactivated mouse or human embryonic fibroblasts in an in vitro fertilisation culture dish. Growth factors potentially involved in primary derivation of hESCs may be lost or diluted in such a system. We established a microdrop method which maintained feeder cells and efficiently generated hESCs. Embryos were donated for stem cell research after fully informed patient consent. A feeder cell layer was made by incubating inactivated mouse embryonic fibroblasts (MEFs) feeder cells in a 50 microl drop of medium (DMEM/10% foetal calf serum) under mineral oil in a small tissue culture dish. MEFs formed a confluent layer and medium was replaced with human embryonic stem medium supplemented with 10% Plasmanate (Bayer) and incubated overnight. Cryopreserved embryos were thawed and cultured until the blastocyst stage and the zona pellucida removed with pronase (2 mg/ml; Calbiochem). A zona-free intact blastocyst was placed in the feeder microdrop and monitored for ES derivation with medium changed every 2-3 d. Proliferating hESCs were passaged into other feeder drops and standard feeder preparation by manual dissection until a stable cell line was established. Six hESC lines (Shef 3-8) were derived. From a total of 46 blastocysts (early to expanded), five hESC lines were generated (Shef 3-7). Shef 3-6 were generated on MEFs from 25 blastocysts. Shef7 was generated on human foetal gonadal embryonic fibroblasts from a further 21 blastocysts. From our experience, microdrop technique is more efficient than conventional method for derivation of hESCs and it is much easier to monitor early hESC derivation. The microdrop method lends itself to good manufacturing practice derivation of hESCs. PMID:20224972

  14. Testing a theory of organizational culture, climate and youth outcomes in child welfare systems: A United States national study

    PubMed Central

    Williams, Nathaniel J.; Glisson, Charles

    2013-01-01

    Theories of organizational culture and climate (OCC) applied to child welfare systems hypothesize that strategic dimensions of organizational culture influence organizational climate and that OCC explains system variance in youth outcomes. This study provides the first structural test of the direct and indirect effects of culture and climate on youth outcomes in a national sample of child welfare systems and isolates specific culture and climate dimensions most associated with youth outcomes. The study applies multilevel path analysis (ML-PA) to a U.S. nationwide sample of 2,380 youth in 73 child welfare systems participating in the second National Survey of Child and Adolescent Well-being. Youths were selected in a national, two-stage, stratified random sample design. Youths’ psychosocial functioning was assessed by caregivers’ responses to the Child Behavior Checklist at intake and at 18-month follow-up. OCC was assessed by front-line caseworkers’ (N=1,740) aggregated responses to the Organizational Social Context measure. Comparison of the a priori and subsequent trimmed models confirmed a reduced model that excluded rigid organizational culture and explained 70% of the system variance in youth outcomes. Controlling for youth- and system-level covariates, systems with more proficient and less resistant organizational cultures exhibited more functional, more engaged, and less stressful climates. Systems with more proficient cultures and more engaged, more functional, and more stressful climates exhibited superior youth outcomes. Findings suggest child welfare administrators can support service effectiveness with interventions that improve specific dimensions of culture and climate. PMID:24094999

  15. Biomass recycle as a means to improve the energy efficiency of CELSS algal culture systems

    NASA Technical Reports Server (NTRS)

    Radmer, R.; Cox, J.; Lieberman, D.; Behrens, P.; Arnett, K.

    1987-01-01

    Algal cultures can be very rapid and efficient means to generate biomass and regenerate the atmosphere for closed environmental life support systems. However, as in the case of most higher plants, a significant fraction of the biomass produced by most algae cannot be directly converted to a useful food product by standard food technology procedures. This waste biomass will serve as an energy drain on the overall system unless it can be efficiently recycled without a significant loss of its energy content. Experiments are reported in which cultures of the alga Scenedesmus obliquus were grown in the light and at the expense of an added carbon source, which either replaced or supplemented the actinic light. As part of these experiments, hydrolyzed waste biomass from these same algae were tested to determine whether the algae themselves could be made part of the biological recycling process. Results indicate that hydrolyzed algal (and plant) biomass can serve as carbon and energy sources for the growth of these algae, suggesting that the efficiency of the closed system could be significantly improved using this recycling process.

  16. Developing New Systems of Data to Advance a Culture of Health

    PubMed Central

    Plough, Alonzo

    2014-01-01

    Introduction: The system dynamics that are driving changes in health and health care in the United States are not well captured by standard data-collection activities. We need data systems that can inform policy and program development and can reflect the increasing recognition that all aspects of people’s lives—their work, families, and communities and not just formal health and health care services—support active and healthy living. Culture of Health Action Areas: Within the Culture of Health (COH) framework, four action areas are further upstream in the sequence of change and point to additional need for more nuanced approaches to data collection: building a shared value of health; fostering collaborations to improve well-being; creating health in more equitable community environments; and transforming health and health care systems. These action areas define the COH framework that we hope will spur research and strategies across sectors to improve health. Conclusion: The action framework for advancing a COH will guide the Robert Wood Johnson Foundation’s (RWJF) grant making and strategic collaborations. It is hoped that the framework will also catalyze changes in how researchers and organizations collect health-related data at all levels. PMID:25995991

  17. Mitigation of climate change impacts by hydrologic and cultural components of traditional acequia irrigation systems

    NASA Astrophysics Data System (ADS)

    Fernald, A.

    2009-12-01

    In northern New Mexico and other physiographically similar semi-arid settings worldwide, traditional irrigation systems divert snowmelt runoff from streams for distribution to valley croplands. This field hydrology and culture study is taking place in three New Mexico watersheds. Ongoing measurements show that seepage to groundwater and subsequent stream recharge from subsurface return flows effectively reduce spring runoff peaks and augment summer baseflow. This retransmission function of traditional acequia irrigated valleys is important for downstream users, particularly in the face of changing climate with projected earlier snowmelt and increased rain. Preliminary evaluations of the community irrigation management structure show high adaptability to climate variation. Water is partitioned to individual users based on water availability, with more water for all in wet years and less for all in dry years. Irrigation water seepage has additional benefits: water quality improvement, wildlife habitat creation, riparian vegetation support, and aesthetic enhancement. Community cohesion and longevity are supported by hydrologic and cultural aspects of the irrigation systems. Lessons learned from these systems promise a window into techniques for sustainable management of linked watersheds and river valleys under future climate change scenarios.

  18. The Connecticut Latino Behavioral Health System: A culturally informed community-academic collaboration.

    PubMed

    Paris, Manuel; Silva, Michelle A; Diaz, Esperanza; Bedregal, Luis E; Cole, Robert A; Añez-Nava, Luis M

    2016-05-01

    The Connecticut Latino Behavioral Health System (LBHS) represents a culturally informed community-academic collaboration that includes agencies focused on mental health, addictions, behavioral health within community health centers, and social rehabilitation; the Yale University Department of Psychiatry; and the Connecticut Department of Mental Health and Addiction Services. The core mission of the LBHS is to expand and enhance the provision of recovery-oriented, and culturally and linguistically appropriate, services to the monolingual Spanish-speaking community in parts of South Central Connecticut. This article outlines the rationale and need for such a collaboration to meet the needs of an underrepresented and underserved ethnic minority group. The process by which these entities came together to develop and successfully implement systemic strategies is described in the context of 2 overarching priorities: (a) workforce development, and (b) access to services. The authors also highlight lessons learned that have informed the decision-making process since the inception of the LBHS, and future directions to ensure that it is prepared to meet changing consumer needs and systemic priorities. (PsycINFO Database Record PMID:27148948

  19. Evaluation of the rat embryo culture system as a predictive test for human teratogens.

    PubMed

    Guest, I; Buttar, H S; Smith, S; Varma, D R

    1994-01-01

    Ingestion of the anticonvulsant drug valproic acid and of the angiotensin converting enzyme inhibitor captopril during pregnancy has been associated with abnormal fetal outcome in humans. In contrast, the use of the antiinflammatory drug ibuprofen and the antihistamine diphenhydramine has not been documented to be embryotoxic in humans. We evaluated the rat embryo culture system as a predictive model of teratogenesis, using these four drugs as test agents. Valproic acid, ibuprofen, and diphenhydramine were embryotoxic, inducing concentration-dependent decreases in growth and a significant increase in anomalies. Valproic acid caused an increase in neural tube defects, ibuprofen increased the incidence of abnormal maxillary processes, and diphenhydramine increased the number of embryos with distorted body morphology. These abnormalities were induced at concentrations of valproic acid and diphenhydramine that are used clinically, but ibuprofen only induced toxicity at concentrations greatly exceeding the therapeutic range. Captopril was not embryotoxic up to 5 mM, the highest concentration tested. These results suggest that the rat embryo culture system produces both false positive and false negative data on the teratogenic potential of drugs. Although such an in vitro assay may be suitable to determine the mechanism of teratogenesis, it is not a sensitive indicator of potential human teratogens on its own. These data support the view that in vitro systems can only supplement clinical and epidemiological observations in humans, possibly as a method to determine mechanisms of actions of teratogens. PMID:8012899

  20. A management system integrating radiation protection and safety supporting safety culture in the hospital.

    PubMed

    Almén, A; Lundh, C

    2015-04-01

    Quality assurance has been identified as an important part of radiation protection and safety for a considerable time period. A rational expansion and improvement of quality assurance is to integrate radiation protection and safety in a management system. The aim of this study was to explore factors influencing the implementing strategy when introducing a management system including radiation protection and safety in hospitals and to outline benefits of such a system. The main experience from developing a management system is that it is possible to create a vast number of common policies and routines for the whole hospital, resulting in a cost-efficient system. One of the key benefits is the involvement of management at all levels, including the hospital director. Furthermore, a transparent system will involve staff throughout the organisation as well. A management system supports a common view on what should be done, who should do it and how the activities are reviewed. An integrated management system for radiation protection and safety includes key elements supporting a safety culture. PMID:25429027

  1. A New Acquisition and Imaging System for Environmental Measurements: An Experience on the Italian Cultural Heritage

    PubMed Central

    Leccese, Fabio; Cagnetti, Marco; Calogero, Andrea; Trinca, Daniele; di Pasquale, Stefano; Giarnetti, Sabino; Cozzella, Lorenzo

    2014-01-01

    A new acquisition system for remote control of wall paintings has been realized and tested in the field. The system measures temperature and atmospheric pressure in an archeological site where a fresco has been put under control. The measuring chain has been designed to be used in unfavorable environments where neither electric power nor telecommunication infrastructures are available. The environmental parameters obtained from the local monitoring are then transferred remotely allowing an easier management by experts in the field of conservation of cultural heritage. The local acquisition system uses an electronic card based on microcontrollers and sends the data to a central unit realized with a Raspberry-Pi. The latter manages a high quality camera to pick up pictures of the fresco. Finally, to realize the remote control at a site not reached by internet signals, a WiMAX connection based on different communication technologies such as WiMAX, Ethernet, GPRS and Satellite, has been set up. PMID:24859030

  2. Engagement in system redesign: A wellness example to enable a cultural transformation.

    PubMed

    Anderson, Bradley; Hansson, Wendy Kai

    2016-09-01

    For the past several years, Interior Health (IH) has worked collaboratively with Aboriginal leaders to build strong relationships and develop an environment of reciprocal accountability and knowledge exchange. All partners are committed to working together to change the healthcare system so that it responds appropriately and effectively to the needs of a population of people with the poorest health outcomes. The development of the IH Aboriginal Health and Wellness Strategy is an example of meaningful engagement with First Nations communities in the IH region at the system level. The strategy was built by IH, led by First Nations, and developed in collaboration with First Nations, Urban, and Métis partners. It aims to address the shared long-term goal of improving the overall health and wellness of Aboriginal people and enable a cultural transformation to overcome the barriers that are stopping Aboriginal people from seeking care from a shared healthcare system. PMID:27576857

  3. A new acquisition and imaging system for environmental measurements: an experience on the Italian cultural heritage.

    PubMed

    Leccese, Fabio; Cagnetti, Marco; Calogero, Andrea; Trinca, Daniele; di Pasquale, Stefano; Giarnetti, Sabino; Cozzella, Lorenzo

    2014-01-01

    A new acquisition system for remote control of wall paintings has been realized and tested in the field. The system measures temperature and atmospheric pressure in an archeological site where a fresco has been put under control. The measuring chain has been designed to be used in unfavorable environments where neither electric power nor telecommunication infrastructures are available. The environmental parameters obtained from the local monitoring are then transferred remotely allowing an easier management by experts in the field of conservation of cultural heritage. The local acquisition system uses an electronic card based on microcontrollers and sends the data to a central unit realized with a Raspberry-Pi. The latter manages a high quality camera to pick up pictures of the fresco. Finally, to realize the remote control at a site not reached by internet signals, a WiMAX connection based on different communication technologies such as WiMAX, Ethernet, GPRS and Satellite, has been set up. PMID:24859030

  4. Documentation of Cultural Heritages Using a GIS Based Information and Management System; Case Study of Safranbolu

    NASA Astrophysics Data System (ADS)

    Seker, D. Z.; Alkan, M.; Kutoglu, S. S.; Akcin, H.

    2010-12-01

    Documentation of the cultural heritage sites is extremely important for monitoring and preserves them from natural disasters and human made activities. Due to its very rich historical background from the first human settlements in Catalhoyuk and Alacahoyuk and civilizations such as Byzantine, Seljuk and Ottoman, there are lots of cultural heritage sites in Turkey. 3D modeling and recording of historical buildings using modern tools and techniques in several locations of Turkey have been conducted and still continuing. The nine cultural sites in Turkey are included in the protection list of UNESCO as cultural heritage and one of them is the township of Safranbolu, which is the one of the most outstanding example of the traditional Turkish Architecture and also unique itself in terms of conservation of the human settlement in their authentic environmental motif up till now. In this study outcomes and further studies of a research project related to study area which is supported by the Turkish National Research Center (TUBITAK) with the project number 106Y157, will be presented in details. The basic aim of the study is development a GIS based information and management system for the city of Safranbolu. All historical buildings which are registered are assigned with the database. 3D modeling some of the selected building among the buildings which are registered as historical monuments using different data comes from different sources similar to their original constructions were realized and then it will be distributed via internet by a web-based information system designed during the project. Also some of the buildings were evaluated using close range photogrammetric technique to obtain their façade reliefs, were also assigned with the database. Designed database consists of 3D models, locations, historical information, cadastral and land register data of the selected buildings together with the other data collected during the project related to buildings. Using this

  5. Automated Static Culture System Cell Module Mixing Protocol and Computational Fluid Dynamics Analysis

    NASA Technical Reports Server (NTRS)

    Kleis, Stanley J.; Truong, Tuan; Goodwin, Thomas J,

    2004-01-01

    This report is a documentation of a fluid dynamic analysis of the proposed Automated Static Culture System (ASCS) cell module mixing protocol. The report consists of a review of some basic fluid dynamics principles appropriate for the mixing of a patch of high oxygen content media into the surrounding media which is initially depleted of oxygen, followed by a computational fluid dynamics (CFD) study of this process for the proposed protocol over a range of the governing parameters. The time histories of oxygen concentration distributions and mechanical shear levels generated are used to characterize the mixing process for different parameter values.

  6. A 3D sphere culture system containing functional polymers for large-scale human pluripotent stem cell production.

    PubMed

    Otsuji, Tomomi G; Bin, Jiang; Yoshimura, Azumi; Tomura, Misayo; Tateyama, Daiki; Minami, Itsunari; Yoshikawa, Yoshihiro; Aiba, Kazuhiro; Heuser, John E; Nishino, Taito; Hasegawa, Kouichi; Nakatsuji, Norio

    2014-05-01

    Utilizing human pluripotent stem cells (hPSCs) in cell-based therapy and drug discovery requires large-scale cell production. However, scaling up conventional adherent cultures presents challenges of maintaining a uniform high quality at low cost. In this regard, suspension cultures are a viable alternative, because they are scalable and do not require adhesion surfaces. 3D culture systems such as bioreactors can be exploited for large-scale production. However, the limitations of current suspension culture methods include spontaneous fusion between cell aggregates and suboptimal passaging methods by dissociation and reaggregation. 3D culture systems that dynamically stir carrier beads or cell aggregates should be refined to reduce shearing forces that damage hPSCs. Here, we report a simple 3D sphere culture system that incorporates mechanical passaging and functional polymers. This setup resolves major problems associated with suspension culture methods and dynamic stirring systems and may be optimal for applications involving large-scale hPSC production. PMID:24936458

  7. An innovative three-dimensional gelatin foam culture system for improved study of glioblastoma stem cell behavior.

    PubMed

    Yang, Meng-Yin; Chiao, Ming-Tsang; Lee, Hsu-Tung; Chen, Chien-Min; Yang, Yi-Chin; Shen, Chiung-Chyi; Ma, Hsin-I

    2015-04-01

    Three-dimensional (3-D) tissue engineered constructs provide a platform for examining how the local extracellular matrix contributes to the malignancy of various cancers, including human glioblastoma multiforme. Here, we describe a simple and innovative 3-D culture environment and assess its potential for use with glioblastoma stem cells (GSCs) to examine the diversification inside the cell mass in the 3-D culture system. The dissociated human GSCs were cultured using gelatin foam. These cells were subsequently identified by immunohistochemical staining, reverse transcriptase-polymerase chain reaction, and Western blot assay. We demonstrate that the gelatin foam provides a suitable microenvironment, as a 3-D culture system, for GSCs to maintain their stemness. The gelatin foam culture system contributes a simplified assessment of cell blocks for immunohistochemistry assay. We show that the significant transcription activity of hypoxia and the protein expression of inflammatory responses are detected at the inside of the cell mass in vitro, while robust expression of PROM1/CD133 and hypoxia-induced factor-1 alpha are detected at the xenografted tumor in vivo. We also examine the common clinical trials under this culture platform and characterized a significant difference of drug resistance. The 3-D gelatin foam culture system can provide a more realistic microenvironment through which to study the in vivo behavior of GSCs to evaluate the role that biophysical factors play in the hypoxia, inflammatory responses and subsequent drug resistance. PMID:24966152

  8. Optimal concentration of hyaluronan and plant protein in different culture systems for in vitro maturation of bovine oocytes.

    PubMed

    Opiela, Jolanta; Latasiewicz, Ewa; Smorag, Zdzisław

    2012-12-01

    With a view to search for optimal concentration of hyaluronan (HA) and plant protein (PP) in different culture systems for in vitro maturation of bovine oocytes, cumulus-oocyte complexes (COCs) were matured in vitro in 2 culture systems (first co-cultured with granulose cells and estrus calf serum (ECS) in 2 mL volume, second without co-culture where ECS was replaced by exogenous hormones and BSA or PP in 100 microL dose under mineral oil). Seven types of media were used; 3 in first system and 4 in second system. To evaluate HA and PP effect on oocytes after in vitro culture an estimation of meiosis stage and a level of DNA fragmentation was performed by TUNEL staining. The highest meiotic maturation (84%) was observed in oocytes cultured in medium enriched with ECS in co-culture with granulose cells (1st system). The lowest meiotic maturation was noted in medium with addition of BSA (43%). The addition of HA in the medium enriched with BSA significantly increased the rate of matured oocytes (67%) and also didn't affect the chromatin quality of individual oocytes. The addition of HA to the culture medium supplemented with a PP decreased the rate of matured oocytes to 54% but no statistical differences were noted. The results of the present study showed that HA supplementation didn't have a detrimental impact on oocyte chromatin integrity and improved bovine oocytes' meiotic maturation in medium supplemented only with BSA without co-culture of granulose cells. PMID:23986966

  9. Development of a Xeno-Free Autologous Culture System for Endothelial Progenitor Cells Derived from Human Umbilical Cord Blood

    PubMed Central

    Park, Soon-Jung; Kim, Hojin; Bae, Daekyeong

    2013-01-01

    Despite promising preclinical outcomes in animal models, a number of challenges remain for human clinical use. In particular, expanding a large number of endothelial progenitor cells (EPCs) in vitro in the absence of animal-derived products is the most critical hurdle remaining to be overcome to ensure the safety and efficiency of human therapy. To develop in vitro culture conditions for EPCs derived from human cord blood (hCB-EPCs), we isolated extracts (UCE) and collagen (UC-collagen) from umbilical cord tissue to replace their animal-derived counterparts. UC-collagen and UCE efficiently supported the attachment and proliferation of hCB-EPCs in a manner comparable to that of animal-derived collagen in the conventional culture system. Our developed autologous culture system maintained the typical characteristics of hCB-EPCs, as represented by the expression of EPC-associated surface markers. In addition, the therapeutic potential of hCB-EPCs was confirmed when the transplantation of hCB-EPCs cultured in this autologous culture system promoted limb salvage in a mouse model of hindlimb ischemia and was shown to contribute to attenuating muscle degeneration and fibrosis. We suggest that the umbilical cord represents a source for autologous biomaterials for the in vitro culture of hCB-EPCs. The main characteristics and therapeutic potential of hCB-EPCs were not compromised in developed autologous culture system. The absence of animal-derived products in our newly developed in vitro culture removes concerns associated with secondary contamination. Thus, we hope that this culture system accelerates the realization of therapeutic applications of autologous hCB-EPCs for human vascular diseases. PMID:24086472

  10. Development of a vessel organ culture system: characterisation of the method and implications for the reduction of animal experiments.

    PubMed

    Zaniboni, Andrea; Zannoni, Augusta; Bernardini, Chiara; De Cecco, Marco; Bombardi, Cristiano; Seren, Eraldo; Forni, Monica; Bacci, Maria Laura

    2013-09-01

    In the field of cardiovascular research, the pig is considered to be an excellent animal model of human diseases. It is well-known that primary cultures of endothelial cells (ECs) are a powerful tool for the study of vascular physiology and pathology, and, according to the principles of the Three Rs, their use results in a substantial reduction in the numbers of experimental animals required. However, a limitation of EC culture is that the cells are not in their physiological context. Here, we describe and characterise a method for the culture of porcine vessels that overcomes the limitation of EC cultures, with the advantage of reducing the number of animals used for research purposes. The organ cultures were set-up by using an aortic cylinder obtained from the arteries of control pigs sacrificed for other experimental purposes. In order to characterise the method, vascular endothelial growth factor (VEGF) secretion, matrix metalloproteinase (MMP) activation and the vessel's structural features were evaluated during organ culture. These analyses confirm that the culture of aortic cylinder lumen, in a medium specific for ECs, results in a stable system in terms of VEGF and MMP secretion. The ECs do not undergo cell division during the organ culture, which is also the case in vivo, if no stimulation occurs. Overall, we show that this novel system closely resembles the in vivo context. Importantly, porcine aortas can be collected from either veterinary surgeries or slaughterhouses, without having to sacrifice animals specifically for the purposes of this type of research. PMID:24168133

  11. Quorum-Sensing Regulation of Constitutive Plantaricin by Lactobacillus plantarum Strains under a Model System for Vegetables and Fruits

    PubMed Central

    Rizzello, Carlo G.; Filannino, Pasquale; Calasso, Maria; Gobbetti, Marco

    2014-01-01

    This study aimed at investigating the regulatory system of bacteriocin synthesis by Lactobacillus plantarum strains in vegetables and fruits in a model system. Sterile and neutralized cell-free supernatant (CFS) from L. plantarum strains grown in MRS broth showed in vitro antimicrobial activities toward various indicator strains. The highest activity was that of L. plantarum C2. The antimicrobial activity was further assayed on vegetable and fruit agar plates (solid conditions) and in juices (liquid conditions). A regulatory mechanism of bacteriocin synthesis via quorum sensing was hypothesized. The synthesis of antimicrobial compounds seemed to be constitutive under solid conditions of growth on vegetable and fruit agar plates. In contrast, it depended on the size of the inoculum when L. plantarum C2 was grown in carrot juice. Only the inoculum of ca. 9.0 log CFU ml−1 produced detectable activity. The genes plnA, plnEF, plnG, and plnH were found in all L. plantarum strains. The genes plnJK and plnN were detected in only three or four strains. Reverse-phase high-performance liquid chromatography purification and mass spectrometry analysis revealed the presence of a mixture of eight peptides in the most active fraction of the CFS from L. plantarum C2. Active peptides were encrypted into bacteriocin precursors, such as plantaricins PlnJ/K and PlnH and PlnG, which are involved in the ABC transport system. A real-time PCR assay showed an increase in the expression of plnJK and plnG during growth of L. plantarum C2 in carrot juice. PMID:24242246

  12. Cell-free protein synthesis systems derived from cultured mammalian cells.

    PubMed

    Brödel, Andreas K; Wüstenhagen, Doreen A; Kubick, Stefan

    2015-01-01

    We present a technology for the production of target proteins using novel cell-free systems derived from cultured human K562 cells and Chinese hamster ovary (CHO) cells. The protocol includes the cultivation of cells, the preparation of translationally active lysates, and the cell-free synthesis of desired proteins. An efficient expression vector based on the internal ribosome entry site (IRES) from the intergenic region (IGR) of the cricket paralysis virus (CrPV) was constructed for both systems. The coupled batch-based platforms enable the synthesis of a broad range of target proteins such as cytosolic proteins, secreted proteins, membrane proteins embedded into endogenous microsomes, and glycoproteins. The glycosylation of erythropoietin demonstrates the successful performance of posttranslational modifications in the novel cell-free systems. Protein yields of approximately 20 μg/ml (K562-based cell-free system) and 50 μg/ml (CHO-based cell-free system) of active firefly luciferase are obtained in the coupled transcription-translation systems within 3 h. As a result, both cell-free protein synthesis systems serve as powerful tools for high-throughput proteomics. PMID:25502197

  13. Diversity of culturable root-associated/endophytic bacteria and their chitinolytic and aflatoxin inhibition activity of peanut plant in China.

    PubMed

    Wang, Kai; Yan, Pei-Sheng; Ding, Qing-Long; Wu, Qin-Xi; Wang, Zhong-Bo; Peng, Jie

    2013-01-01

    A total of 72 isolates of root-associated/endophytic (RAE) bacteria were isolated from peanut plants grown in the main producing areas of 6 provinces in China. The 16S rRNA gene sequences of these isolates were determined and phylogenetic analyses revealed that 72 isolates belonged to the classes Bacilli (49 isolates) and Gammaproteobacteria (23 isolates). The majority of RAE bacteria in Bacilli belonged to 2 genera, Bacillus and Lysinibacillus (48 and 1) while those in Gammaproteobacteria belonged to the genera Enterobacter, Serratia, Stenotrophomonas, and Pseudomonas (7, 11, 3 and 2 isolates, respectively). This is the first report of Lysinibacillus xylanilyticus isolate as biocontrol agent against AFs. All of the selected RAE bacteria showed inhibitory activities against Aspergillus parasiticus (A. parasiticus) growth and/or aflatoxins (AFs) production by visual agar plate assay and tip culture method. Most of the RAE bacteria strains (96 % strains) were determined to have decreased mycelia growth or AFs production levels by >50 % (p < 0.05). Bacterial isolates were further characterized for chitinolytic activity and 22 strains (30 % strains) of identified RAE bacteria degraded colloidal chitin on the chitin medium plate. Ten selected chitinolytic RAE bacteria were tested for antifungal activity on peanuts and most of them significantly decreased mycelial growth and AFs production levels by >90 %. These results showed a wide distribution of biological control bacteria against AFs in Chinese peanut main producing areas and the selected RAE bacteria could potentially be utilized for the biocontrol of toxicogenic fungi. PMID:23108663

  14. Isolation of culturable aerobic bacteria and evidence of Kerstersia gyiorum from the blowhole of captive Yangtze finless porpoises.

    PubMed

    Wan, Xiaoling; McLaughlin, Richard William; Zhou, Junying; Hao, Yujiang; Zheng, Jinsong; Wang, Ding

    2016-08-01

    Bacterial respiratory illnesses are problematic in aquatic mammals such as the Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis; YFP), which is now at a critically endangered status. Yet little is known about the bacteria inhabiting the respiratory tract of YFPs. In this study, we preliminarily characterized the culturable aerobic bacteria from blow samples of captive YFPs. The bacterial diversity was assessed through cultivation by direct exhalation onto Columbia blood agar plates and identification of representative isolates through 16S rRNA gene sequence analysis. In total, eleven bacterial species belonging to four phyla Proteobacteria (71 %), Firmicutes (25 %), Bacteroidetes (3 %) and Actinobacteria (1 %) were identified. Most of these isolates were opportunistic pathogens found in respiratory illnesses in humans and animals. We also reported the first case of Kerstersia gyiorum isolated from an animal. This work provides a preliminary assessment of the bacteria present in the respiratory tract of captive YFPs, which will be an important first step in elucidating the roles of normal microbiota in maintaining respiratory health of YFPs. This study also points out the necessity of future long-term monitoring of blowhole microorganisms in the YFPs and making emergency preparedness plans for respiratory tract infections. These measures can aid in assessing the pathogenic risk of the critically endangered YFP populations. PMID:27251558

  15. Interactions between Skeletal Muscle Myoblasts and their Extracellular Matrix Revealed by a Serum Free Culture System

    PubMed Central

    Chaturvedi, Vishal; Dye, Danielle E.; Kinnear, Beverley F.; van Kuppevelt, Toin H.; Grounds, Miranda D.; Coombe, Deirdre R.

    2015-01-01

    Decellularisation of skeletal muscle provides a system to study the interactions of myoblasts with muscle extracellular matrix (ECM). This study describes the efficient decellularisation of quadriceps muscle with the retention of matrix components and the use of this matrix for myoblast proliferation and differentiation under serum free culture conditions. Three decellularisation approaches were examined; the most effective was phospholipase A2 treatment, which removed cellular material while maximizing the retention of ECM components. Decellularised muscle matrices were then solubilized and used as substrates for C2C12 mouse myoblast serum free cultures. The muscle matrix supported myoblast proliferation and differentiation equally as well as collagen and fibronectin. Immunofluorescence analyses revealed that myoblasts seeded on muscle matrix and fibronectin differentiated to form long, well-aligned myotubes, while myoblasts seeded on collagen were less organized. qPCR analyses showed a time dependent increase in genes involved in skeletal muscle differentiation and suggested that muscle-derived matrix may stimulate an increased rate of differentiation compared to collagen and fibronectin. Decellularized whole muscle three-dimensional scaffolds also supported cell adhesion and spreading, with myoblasts aligning along specific tracts of matrix proteins within the scaffolds. Thus, under serum free conditions, intact acellular muscle matrices provided cues to direct myoblast adhesion and migration. In addition, myoblasts were shown to rapidly secrete and organise their own matrix glycoproteins to create a localized ECM microenvironment. This serum free culture system has revealed that the correct muscle ECM facilitates more rapid cell organisation and differentiation than single matrix glycoprotein substrates. PMID:26030912

  16. Expression System Based on an MTIIa Promoter to Produce hPSA in Mammalian Cell Cultures.

    PubMed

    Santos, Anderson K; Parreira, Ricardo C; Resende, Rodrigo R

    2016-01-01

    Because of the limitations of standard culture techniques, the development of new recombinant protein expression systems with biotechnological potential is a key challenge. Ideally, such systems should be able to effectively and accurately synthesize a protein of interest with intrinsic metabolic capacity. Here, we describe such a system that was designed based on a plasmid vector containing promoter elements derived from the metallothionein MTIIa promoter, as well as processing and purification elements. This promoter can be induced by heavy metals in a culture medium to induce the synthesis of human prostate-specific antigen (hPSA), which has been modified to insert elements for purification, proteolysis, and secretion. We optimized hPSA production in this system by comparing the effects and contributions of ZnCl2, CdCl2, and CuSO4 in HEK293FT, HeLa, BHK-21, and CHO-K1 cells. We also compared the effectiveness of three different transfection agents: multi-walled carbon nanotubes, Lipofectamine 2000, and X-tremeGENE HP Reagent. hPSA production was confirmed via the detection of enhanced green fluorescent protein fluorescence, and cell viability was determined. The expression of hPSA was compared with that of the native protein produced by LNCaP cells, using enzyme-linked immunosorbent assay and sodium dodecyl sulfate polyacrylamide gel electrophoresis. X-tremeGENE reagent, the BHK-21 cell line, and CuSO4 showed the highest hPSA production rates. Furthermore, BHK-21 cells were more resistant to the oxidative stress caused by 100 μM CuSO4. These results suggest that the proposed optimized inducible expression system can effectively produce recombinant proteins with desired characteristics for a wide range of applications in molecular biology. PMID:27582737

  17. Expression System Based on an MTIIa Promoter to Produce hPSA in Mammalian Cell Cultures

    PubMed Central

    Santos, Anderson K.; Parreira, Ricardo C.; Resende, Rodrigo R.

    2016-01-01

    Because of the limitations of standard culture techniques, the development of new recombinant protein expression systems with biotechnological potential is a key challenge. Ideally, such systems should be able to effectively and accurately synthesize a protein of interest with intrinsic metabolic capacity. Here, we describe such a system that was designed based on a plasmid vector containing promoter elements derived from the metallothionein MTIIa promoter, as well as processing and purification elements. This promoter can be induced by heavy metals in a culture medium to induce the synthesis of human prostate-specific antigen (hPSA), which has been modified to insert elements for purification, proteolysis, and secretion. We optimized hPSA production in this system by comparing the effects and contributions of ZnCl2, CdCl2, and CuSO4 in HEK293FT, HeLa, BHK-21, and CHO-K1 cells. We also compared the effectiveness of three different transfection agents: multi-walled carbon nanotubes, Lipofectamine 2000, and X-tremeGENE HP Reagent. hPSA production was confirmed via the detection of enhanced green fluorescent protein fluorescence, and cell viability was determined. The expression of hPSA was compared with that of the native protein produced by LNCaP cells, using enzyme-linked immunosorbent assay and sodium dodecyl sulfate polyacrylamide gel electrophoresis. X-tremeGENE reagent, the BHK-21 cell line, and CuSO4 showed the highest hPSA production rates. Furthermore, BHK-21 cells were more resistant to the oxidative stress caused by 100 μM CuSO4. These results suggest that the proposed optimized inducible expression system can effectively produce recombinant proteins with desired characteristics for a wide range of applications in molecular biology. PMID:27582737

  18. Cultures in hospitals and their influence on attitudes to, and satisfaction with, the use of clinical information systems.

    PubMed

    Callen, Joanne L; Braithwaite, Jeffrey; Westbrook, Johanna I

    2007-08-01

    Studies have identified the importance of hospitals' organisational culture in clinical information system diffusion, but few have quantified its role. This study measured organisational culture in two Australian hospitals to explore whether clinicians' perceptions of culture related to their attitudes to, and satisfaction with, a computerised provider order entry system (CPOE). Data were collected using the Organisational Culture Inventory and a user-satisfaction survey administered to a population of 249 clinicians from the two hospitals. One hospital used CPOE to order clinical laboratory and radiology tests and view results, and the other used the test viewing function only, the ordering facility being planned for later implementation. We found a relationship between culture and clinicians' attitudes, with those in the constructive culture hospital more likely to express positive attitudes towards CPOE, whereas those in the aggressive/defensive hospital were more likely to be negative. The relationship between culture and attitudes towards clinical information systems should be taken into account when planning for their adoption. PMID:17475380

  19. Development of an Integrated Microfluidic Perfusion Cell Culture System for Real-Time Microscopic Observation of Biological Cells

    PubMed Central

    Lin, Lung; Wang, Shih-Siou; Wu, Min-Hsien; Oh-Yang, Chih-Chin

    2011-01-01

    This study reports an integrated microfluidic perfusion cell culture system consisting of a microfluidic cell culture chip, and an indium tin oxide (ITO) glass-based microheater chip for micro-scale perfusion cell culture, and its real-time microscopic observation. The system features in maintaining both uniform, and stable chemical or thermal environments, and providing a backflow-free medium pumping, and a precise thermal control functions. In this work, the performance of the medium pumping scheme, and the ITO glass microheater were experimentally evaluated. Results show that the medium delivery mechanism was able to provide pumping rates ranging from 15.4 to 120.0 μL·min−1. In addition, numerical simulation and experimental evaluation were conducted to verify that the ITO glass microheater was capable of providing a spatially uniform thermal environment, and precise temperature control with a mild variation of ±0.3 °C. Furthermore, a perfusion cell culture was successfully demonstrated, showing the cultured cells were kept at high cell viability of 95 ± 2%. In the process, the cultured chondrocytes can be clearly visualized microscopically. As a whole, the proposed cell culture system has paved an alternative route to carry out real-time microscopic observation of biological cells in a simple, user-friendly, and low cost manner. PMID:22164082

  20. The use of astrocytes in culture as model systems for evaluating neurotoxic-induced-injury.

    PubMed

    Aschner, M; Kimelberg, H K

    1991-01-01

    The prevailing thought that astrocytes function predominantly as passive metabolic or even physical support for neurons has faded over the last 20 years. Today these stellar shaped cells are credited with an expanded role, playing key functions in CNS development, homeostasis, and pathology. In probing their expanded roles, primary astrocyte culture systems have proven to be an indispensable tool. Astrocytes have been implicated in both a defensive and facilitatory capacity for many toxic injuries. Evidence for a protective role of astrocytes in modulating CNS toxicity is afforded by observations that the toxicity of glutamate to cortical neurons is diminished upon astrocytic enrichment of the cell culture (Rosenberg and Aizenman, 1989). In cultures of rat cerebral cortex in which astrocyte proliferation is stringently suppressed, glutamate neurotoxicity occurs at low glutamate concentrations similar to those which are normally found in the extracellular space in the hippocampus. In the presence of excess astrocytes, concentrations of glutamate one-hundred fold higher are required to produce equivalent neurotoxicity (Rosenberg and Aizenman, 1989). Astrocytes can facilitate the action of neurotoxins via a modulating process which takes place within the astrocyte or by a direct cytotoxic effect. Whereas primary astrocyte cultures remain unaffected by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP; Marini et al., 1989), they function prominently in the selective destruction of dopaminergic neurons of the nigrostriatal pathway in humans, other primates and rodents (Davis et al. 1979; Langston et al., 1983; Burns et al., 1983; Langston et al., 1984; Heikkila et al., 1984; Jarvis and Wagner, 1985). Thus, while MPTP by itself is not toxic to cerebellar cells in co-culture with cerebellar astrocytes, MPTP is toxic to the granule cells (Marini et al, 1989). This is thought to be due to an astrocyte-mediated conversion of MPTP to its highly polar and toxic metabolite, 1

  1. Effect of Motility on Surface Colonization and Reproductive Success of Pseudomonas fluorescens in Dual-Dilution Continuous Culture and Batch Culture Systems

    PubMed Central

    Korber, Darren R.; Lawrence, John R.; Caldwell, Douglas E.

    1994-01-01

    The colonization of glass surfaces by motile and nonmotile strains of Pseudomonas fluorescens was evaluated by using dual-dilution continuous culture (DDCC), competitive and noncompetitive attachment assays, and continuous-flow slide culture. Both strains possessed identical growth rates whether in the attached or planktonic state. Results of attachment assays using radiolabeled bacteria indicated that both strains obeyed first-order (monolayer) adsorption kinetics in pure culture. However, the motile strain attached about four times more rapidly and achieved higher final cell densities on surfaces than did the nonmotile strain (2.03 × 108 versus 5.57 × 107 cells vial-1) whether evaluated alone or in cocultures containing motile and nonmotile P. fluorescens. These kinetics were attributed to the increased transport of motile cells from the bulk aqueous phase to the hydrodynamic boundary layer where bacterial attachment, growth, and recolonization could occur. First-order attachment kinetics were also observed for both strains by using continuous-flow slide culture assays analyzed by image analysis. The DDCC system contained both aqueous and particulate phases which could be diluted independently. DDCC results indicated that when cocultures containing motile and nonmotile P. fluorescens colonized solid particles, the motile strain replaced the nonmotile strain in the system over time. Increasing the aqueous-phase rates of dilution decreased the time required for extinction of the nonmotile strain while concurrently decreasing the overall carrying capacity of the DDCC system for both strains. These results confirmed that bacterial motility conveyed a selective advantage during surface colonization even in aqueous-phase systems not dominated by laminar flow. PMID:16349247

  2. Synchronization of stochastic systems: from paddlefish electroreceptors to human epileptic glial cell cultures

    NASA Astrophysics Data System (ADS)

    Neiman, Alexander

    2000-03-01

    Synchronization is one of the fundamental nonlinear phenomena observed in nature. We have studied stochastic synchronization in the electrosensitive system of the paddlefish, Polyodon spathula and have also applied synchronization analysis to networks of glial cells cultured from brain tissue of patients with severe epilepsy. We also present theoretical and numerical models for stochastic synchronization. The electrosensitive system of the paddlefish consists of tens of thousands of electroreceptors located mainly on the "rostrum", which serves as an antenna to locate plankton. Each electroreceptor is a noisy oscillator with natural frequencies in the range of 30-90 Hz. We study synchronization in vivo due to 3-20 Hz external periodic electric fields, which correspond to natural signals produced by Daphnia, the usual prey of paddlefish. We find that for signals whose strengths are in the range that paddlefish customarily encounter in the wild, synchronization coding offers a plausible alternative to the more usual rate coding. We also have studied mutual synchronization between different electroreceptors. Although the spontaneous firing of distant electroreceptors is not synchronized, synchronization is observed when external periodic or even noisy electric fields are applied. We have applied the same analysis techniques to examine synchronization between groups of glial cells. In contrast to cultures of healthy astrocytes, which demonstrate calcium waves, the networks from epileptic tissue are characterized by spatially disordered hyper activity. Nevertheless, we have found that, in many cases, synchronized activity is a rather typical for tissue taken from the uncus region of the brain.

  3. Caged Neuron MEA: A system for long-term investigation of cultured neural network connectivity

    PubMed Central

    Erickson, Jonathan; Tooker, Angela; Tai, Y-C.; Pine, Jerome

    2008-01-01

    Traditional techniques for investigating cultured neural networks, such as the patch clamp and multi-electrode array, are limited by: 1) the number of identified cells which can be simultaneously electrically contacted, 2) the length of time for which cells can be studied, and 3) the lack of one-to-one neuron-to-electrode specificity. Here, we present a new device—the caged neuron multi-electrode array—which overcomes these limitations. This micro-machined device consists of an array of neurocages which mechanically trap a neuron near an extracellular electrode. While the cell body is trapped, the axon and dendrites can freely grow into the surrounding area to form a network. The electrode is bi-directional, capable of both stimulating and recording action potentials. This system is non-invasive, so that all constituent neurons of a network can be studied over its lifetime with stable one-to-one neuron-to-electrode correspondence. Proof-of-concept experiments are described to illustrate that functional networks form in a neurochip system of 16 cages in a 4×4 array, and that suprathreshold connectivity can be fully mapped over several weeks. The neurochip opens a new domain in neurobiology for studying small cultured neural networks. PMID:18775453

  4. Utility of poly(ethylene glycol) based hydrogels for improved neural cell culture and cell delivery systems

    NASA Astrophysics Data System (ADS)

    Namba, Rachael

    Cell transplant therapy is a very promising treatment for patients afflicted with neurodegenerative diseases and involves replacing dying neurons with fresh fetal tissue that is composed of both new neurons and multipotent neural precursor cells (NPCs). Unfortunately, this therapy is not clinically feasible given the scarcity of fresh donor fetal tissue. The NPCs present in fetal tissue may be a promising renewable cell source because they can be instructed to divide indefinitely in culture, expanding the number of NPCs that are available. Importantly, NPCs can also be instructed to differentiate into the therapeutic neurons needed to treat patients. However, when NPCs are expanded in culture, they increasingly differentiate into glial cells. Glial cell contamination not only introduces many uncharacterized signals into NSC cultures, but when transplanted into the brain, glia can contribute to inflammation and glial scarring. The goal of this project is to identify in vitro culture conditions that can be used to expand fresh NPCs while completely avoiding glial differentiation, only allowing the generation of neuronal progeny with therapeutic potential. The culture system used in this work was chosen carefully, and after exploring the utility of standard neural cell culture systems such as monolayer, neurosphere, and three-dimensional fibrin matrices, an advantageous culture system was identified. The chosen culture system is a synthetic, degradable, photo-polymerizable, poly (ethylene) glycocl (PEG) hydrogel. This thesis experimentally demonstrates advantages of PEG hydrogel culture over the other culture systems including enrichment of the initial cell population for NPCs, clonal expansion capabilities, increased potency of proliferation-stimulating mitogens, decreased reactivity and growth of glial cells, and control over the initiation and orientation of neural process growth. PEG hydrogels were used to directly assess the influence of various ECM proteins and

  5. Evaluation of a carp primary hepatocyte culture system for screening chemicals for oestrogenic activity.

    PubMed

    Bickley, L K; Lange, A; Winter, M J; Tyler, C R

    2009-09-14

    variation observed in the vitellogenic response between different cultures, data from the different donor fish could be compared by standardising responses relative to the maximum VTG induction in each culture following exposure to E2. Adopting this approach in the future will allow for data from different hepatocyte cultures and from donor fish of different sexes, age and stage of maturity to be compared with greater consistency. Measurement of vtg mRNA expression was relatively more sensitive to the oestrogenic effects of E2 exposure than measurement of VTG protein (the LOEC at the transcriptome level was 10-fold lower [0.01 microM E2] than at the protein level [0.1 microM E2]) and changes in vtg mRNA expression showed less variation between individual hepatocyte isolations. Measurement of vtg mRNA in the hepatocyte culture system therefore may offer the most sensitive and consistent option for the screening of chemicals with oestrogenic activity in fish primary hepatocyte cultures. PMID:19695720

  6. Establishment of a Novel Lingual Organoid Culture System: Generation of Organoids Having Mature Keratinized Epithelium from Adult Epithelial Stem Cells

    NASA Astrophysics Data System (ADS)

    Hisha, Hiroko; Tanaka, Toshihiro; Kanno, Shohei; Tokuyama, Yoko; Komai, Yoshihiro; Ohe, Shuichi; Yanai, Hirotsugu; Omachi, Taichi; Ueno, Hiroo

    2013-11-01

    Despite the strong need for the establishment of a lingual epithelial cell culture system, a simple and convenient culture method has not yet been established. Here, we report the establishment of a novel lingual epithelium organoid culture system using a three-dimensional matrix and growth factors. Histological analyses showed that the generated organoids had both a stratified squamous epithelial cell layer and a stratum corneum. Very recently, we showed via a multicolor lineage tracing method that Bmi1-positive stem cells exist at the base of the epithelial basal layer in the interpapillary pit. Using our new culture system, we found that organoids could be generated by single Bmi1-positive stem cells and that in the established organoids, multiple Bmi1-positive stem cells were generated at the outermost layer. Moreover, we observed that organoids harvested at an early point in culture could be engrafted and maturate in the tongue of recipient mice and that the organoids generated from carcinogen-treated mice had an abnormal morphology. Thus, this culture system presents valuable settings for studying not only the regulatory mechanisms of lingual epithelium but also lingual regeneration and carcinogenesis.

  7. Gelatine methacrylamide-based hydrogels: an alternative three-dimensional cancer cell culture system.

    PubMed

    Kaemmerer, Elke; Melchels, Ferry P W; Holzapfel, Boris M; Meckel, Tobias; Hutmacher, Dietmar W; Loessner, Daniela

    2014-06-01

    Modern cancer research requires physiological, three-dimensional (3-D) cell culture platforms, wherein the physical and chemical characteristics of the extracellular matrix (ECM) can be modified. In this study, gelatine methacrylamide (GelMA)-based hydrogels were characterized and established as in vitro and in vivo spheroid-based models for ovarian cancer, reflecting the advanced disease stage of patients, with accumulation of multicellular spheroids in the tumour fluid (ascites). Polymer concentration (2.5-7% w/v) strongly influenced hydrogel stiffness (0.5±0.2kPa to 9.0±1.8kPa) but had little effect on solute diffusion. The diffusion coefficient of 70kDa fluorescein isothiocyanate (FITC)-labelled dextran in 7% GelMA-based hydrogels was only 2.3 times slower compared to water. Hydrogels of medium concentration (5% w/v GelMA) and stiffness (3.4kPa) allowed spheroid formation and high proliferation and metabolic rates. The inhibition of matrix metalloproteinases and consequently ECM degradability reduced spheroid formation and proliferation rates. The incorporation of the ECM components laminin-411 and hyaluronic acid further stimulated spheroid growth within GelMA-based hydrogels. The feasibility of pre-cultured GelMA-based hydrogels as spheroid carriers within an ovarian cancer animal model was proven and led to tumour development and metastasis. These tumours were sensitive to treatment with the anti-cancer drug paclitaxel, but not the integrin antagonist ATN-161. While paclitaxel and its combination with ATN-161 resulted in a treatment response of 33-37.8%, ATN-161 alone had no effect on tumour growth and peritoneal spread. The semi-synthetic biomaterial GelMA combines relevant natural cues with tunable properties, providing an alternative, bioengineered 3-D cancer cell culture in in vitro and in vivo model systems. PMID:24590158

  8. Enhanced removal of petroleum hydrocarbons using a bioelectrochemical remediation system with pre-cultured anodes.

    PubMed

    Venkidusamy, Krishnaveni; Megharaj, Mallavarapu; Marzorati, Massimo; Lockington, Robin; Naidu, Ravi

    2016-01-01

    Bioelectrochemical remediation (BER) systems such as microbial fuel cells (MFCs) have recently emerged as a green technology for the effective remediation of petroleum hydrocarbon contaminants (PH) coupled with simultaneous energy recovery. Recent research has shown that biofilms previously enriched for substrate degrading bacteria resulted in excellent performance in terms of substrate removal and electricity generation but the effects on hydrocarbon contaminant degradation were not examined. Here we investigate the differences between enriched biofilm anodes and freshly inoculated new anodes in diesel fed single chamber mediatorless microbial fuel cells (DMFC) using various techniques for the enhancement of PH contaminant remediation with concomitant electricity generation. An anodophilic microbial consortium previously selected for over a year through continuous culturing with a diesel concentration of about 800mgl(-1) and which now showed complete removal of this concentration of diesel within 30days was compared to that of a freshly inoculated new anode MFC (showing 83.4% removal of diesel) with a simultaneous power generation of 90.81mW/m(2) and 15.04mW/m(2) respectively. The behaviour of pre-cultured anodes at a higher concentration of PH (8000mgl(-1)) was also investigated. Scanning electron microscopy observation revealed a thick biofilm covering the pre-cultured anodic electrode but not the anode from the freshly inoculated MFC. High resolution imaging showed the presence of thin 60nm diametre pilus-like projections emanating from the cells. Anodic microbial community profiling confirmed that the selection for diesel degrading exoelectrogenic bacteria had occurred. Identification of a biodegradative gene (alkB) provided strong evidence of the catabolic pathway used for diesel degradation in the DMFCs. PMID:26360455

  9. High prevalence of Kingella kingae in joint fluid from children with septic arthritis revealed by the BACTEC blood culture system.

    PubMed Central

    Yagupsky, P; Dagan, R; Howard, C W; Einhorn, M; Kassis, I; Simu, A

    1992-01-01

    In an effort to improve detection of fastidious organisms, joint fluid aspirates of pediatric patients were inoculated into BACTEC 460 aerobic blood culture bottles, in addition to cultures on solid media. Culture records for the 1988 to 1991 period were reviewed to compare the performance of both methods for the recovery of pathogens. Overall, 216 children underwent a diagnostic joint tap, and 63 specimens grew significant organisms, including Kingella kingae in 14. While both methods were comparable for recovery of usual pathogens, with a single exception, K. kingae isolates were detected by the BACTEC system only. K. kingae appears to be a more common cause of septic arthritis in children than has been previously recognized. The BACTEC blood culture system enhances the recovery of K. kingae from joint fluid and improves bacteriologic diagnosis of pediatric septic arthritis. PMID:1583131

  10. Evaluation of Interferon Resistance in Newly Established Genotype 1b Hepatitis C Virus Cell Culture System

    PubMed Central

    Taniguchi, Miki; Tasaka-Fujita, Megumi; Nakagawa, Mina; Watanabe, Takako; Kawai-Kitahata, Fukiko; Otani, Satoshi; Goto, Fumio; Nagata, Hiroko; Kaneko, Shun; Nitta, Sayuri; Murakawa, Miyako; Nishimura-Sakurai, Yuki; Azuma, Seishin; Itsui, Yasuhiro; Mori, Kenichi; Yagi, Shintaro; Kakinuma, Sei; Asahina, Yasuhiro; Watanabe, Mamoru

    2016-01-01

    Background and Aims: The hepatitis C virus (HCV) genotype 1b is known to exhibit treatment resistance with respect to interferon (IFN) therapy. Substitution of amino acids 70 and 91 in the core region of the 1b genotype is a significant predictor of liver carcinogenesis and poor response to pegylated-IFN-α and ribavirin therapy. However, the molecular mechanism has not yet been clearly elucidated because of limitations of the HCV genotype 1b infectious model. Recently, the TPF1-M170T HCV genotype 1b cell culture system was established, in which the clone successfully replicates and infects Huh-7-derived Huh7-ALS32.50 cells. Therefore, the purpose of this study was to compare IFN resistance in various HCV clones using this system. Methods: HCV core amino acid substitutions R70Q and L91M were introduced to the TPF1-M170T clone and then transfected into Huh7-ALS32.50 cells. To evaluate the production of each virus, intracellular HCV core antigens were measured. Results were confirmed with Western blot analysis using anti-NS5A antibodies, and IFN sensitivity was subsequently measured. Results: Each clone was transfected successfully compared with JFH-1, with a significant difference in intracellular HCV core antigen (p < 0.05), an indicator of continuous HCV replication. Among all clones, L91M showed the highest increase in the HCV core antigen and HCV protein. There was no significant resistance against IFN treatment in core substitutions; however, IFN sensitivity was significantly different between the wildtype core and JFH-1 (p < 0.05). Conclusions: A novel genotype 1b HCV cell culture was constructed with core amino acid substitutions, which demonstrated IFN resistance of genotype 1b. This system will be useful for future analyses into the mechanisms of HCV genotype 1b treatment. PMID:27047766

  11. Real-time quantitative fluorescence measurement of microscale cell culture analog systems

    NASA Astrophysics Data System (ADS)

    Oh, Taek-il; Kim, Donghyun; Tatosian, Daniel; Sung, Jong Hwan; Shuler, Michael

    2007-02-01

    A microscale cell culture analog (μCCA) is a cell-based lab-on-a-chip assay that, as an animal surrogate, is applied to pharmacological studies for toxicology tests. A μCCA typically comprises multiple chambers and microfluidics that connect the chambers, which represent animal organs and blood flow to mimic animal metabolism more realistically. A μCCA is expected to provide a tool for high-throughput drug discovery. Previously, a portable fluorescence detection system was investigated for a single μCCA device in real-time. In this study, we present a fluorescence-based imaging system that provides quantitative real-time data of the metabolic interactions in μCCAs with an emphasis on measuring multiple μCCA samples simultaneously for high-throughput screening. The detection system is based on discrete optics components, with a high-power LED and a charge-coupled device (CCD) camera as a light source and a detector, for monitoring cellular status on the chambers of each μCCA sample. Multiple samples are characterized mechanically on a motorized linear stage, which is fully-automated. Each μCCA sample has four chambers, where cell lines MES-SA/DX- 5, and MES-SA (tumor cells of human uterus) have been cultured. All cell-lines have been transfected to express the fusion protein H2B-GFP, which is a human histone protein fused at the amino terminus to EGFP. As a model cytotoxic drug, 10 μM doxorubicin (DOX) was used. Real-time quantitative data of the intensity loss of enhanced green fluorescent protein (EGFP) during cell death of target cells have been collected over several minutes to 40 hours. Design issues and improvements are also discussed.

  12. An Axenic Plant Culture System for Optimal Growth in Long-Term Studies: Design and Maintenance

    NASA Technical Reports Server (NTRS)

    Henry, Amelia; Doucette, William; Norton, Jeanette; Jones, Scott; Chard, Julie; Bugbee, Bruce

    2006-01-01

    The symbiotic co-evolution of plants and microbes leads to difficulties in understanding which of the two components is responsible for a given environmental response. Plant-microbe studies greatly benefit from the ability to grow plants in axenic (sterile) culture. Several studies have used axenic plant culture systems, but experimental procedures are often poorly documented, the plant growth environment is not optimal, and axenic conditions are not rigorously verified. We developed a unique axenic system using inert components that promotes plant health and can be kept sterile for at least 70 d. Crested wheatgrass (Agropyron cristatum cv. DII) plants were grown in sand within flow-through glass columns that were positively pressured with filtered air. Plant health was optimized by regulating temperature, light level, CO2 concentration, humidity, and nutrients. The design incorporates several novel aspects, such as pretreatment of the sand with Fe, graduated sand layers to optimize the air-water balance of the root zone, and modification of a laminar flow hood to serve as a plant growth chamber. Adaptations of several sterile techniques were necessary for maintenance of axenic conditions. Axenic conditions were verified by plating and staining leachates as well as rhizoplane stain. This system was designed to study nutrient and water stress effects on root exudates, but is useful for assessing a broad range of plant-microbe-environment interactions. Based on total organic C analysis, 74% of exudates was recovered in the leachate, 6% was recovered in the bulk sand, and 17% was recovered in the rhizosphere sand. Carbon in the leachate after 70 d reached 255 micro-g/d. Fumaric, malic, malonic, oxalic, and succinic acids were measured as components of the root exudates.

  13. A Winogradsky-based culture system shows an association between microbial fermentation and cystic fibrosis exacerbation

    PubMed Central

    Quinn, Robert A; Whiteson, Katrine; Lim, Yan-Wei; Salamon, Peter; Bailey, Barbara; Mienardi, Simone; Sanchez, Savannah E; Blake, Don; Conrad, Doug; Rohwer, Forest

    2015-01-01

    There is a poor understanding of how the physiology of polymicrobial communities in cystic fibrosis (CF) lungs contributes to pulmonary exacerbations and lung function decline. In this study, a microbial culture system based on the principles of the Winogradsky column (WinCF system) was developed to study the physiology of CF microbes. The system used glass capillary tubes filled with artificial sputum medium to mimic a clogged airway bronchiole. Chemical indicators were added to observe microbial physiology within the tubes. Characterization of sputum samples from seven patients showed variation in pH, respiration, biofilm formation and gas production, indicating that the physiology of CF microbial communities varied among patients. Incubation of homogenized tissues from an explant CF lung mirrored responses of a Pseudomonas aeruginosa pure culture, supporting evidence that end-stage lungs are dominated by this pathogen. Longitudinal sputum samples taken through two exacerbation events in a single patient showed that a two-unit drop in pH and a 30% increase in gas production occurred in the tubes prior to exacerbation, which was reversed with antibiotic treatment. Microbial community profiles obtained through amplification and sequencing of the 16S rRNA gene showed that fermentative anaerobes became more abundant during exacerbation and were then reduced during treatment where P. aeruginosa became the dominant bacterium. Results from the WinCF experiments support the model where two functionally different CF microbial communities exist, the persistent Climax Community and the acute Attack Community. Fermentative anaerobes are hypothesized to be the core members of the Attack Community and production of acidic and gaseous products from fermentation may drive developing exacerbations. Treatment targeting the Attack Community may better resolve exacerbations and resulting lung damage. PMID:25514533

  14. Controlled evaluation of the agar-slide and radiometric blood culture systems for the detection of bacteremia and fungemia.

    PubMed Central

    Weinstein, M P; Reller, L B; Mirrett, S; Stratton, C W; Reimer, L G; Wang, W L

    1986-01-01

    A commercially available agar-slide blood culture bottle (Septi-Chek; Roche Diagnostics, Div. Hoffman-La Roche, Inc., Nutley, N.J.) was compared with the radiometric blood culture system (BACTEC; Johnston Laboratories, Inc., Towson, Md.) in 8,544 paired blood cultures from adult patients. The systems were inoculated with equal volumes (10 ml) of blood. Overall, there was no statistically significant difference between the two systems in the recovery of clinically important microorganisms, but significantly more members of the family Enterobacteriaceae other than Escherichia coli were detected by the agar-slide system (P less than 0.005). The agar-slide system detected more fungi, and the BACTEC detected more anaerobic bacteria; however, small numbers of recovered organisms precluded statistical significance. When microorganisms grew in both systems, their presence was detected one or more days earlier in the BACTEC (P less than 0.001). More contaminants grew in the agar-slide system (P less than 0.001). Both systems performed well, and either system should provide high yield and prompt detection of positive blood cultures in patients with bacteremia and fungemia if used in an optimal way as recommended by the respective manufacturers. PMID:3517047

  15. The Pharmaceutical Aerosol Deposition Device on Cell Cultures (PADDOCC) in vitro system: design and experimental protocol.

    PubMed

    Hein, Stephanie; Bur, Michael; Kolb, Tobias; Muellinger, Bernhard; Schaefer, Ulrich F; Lehr, Claus-Michael

    2010-08-01

    The development of aerosol medicines typically involves numerous tests on animals, due to the lack of adequate in vitro models. A new in vitro method for testing pharmaceutical aerosol formulations on cell cultures was developed, consisting of an aerosolisation unit fitting a commercial dry powder inhaler (HandiHaler(c), Boehringer Ingelheim, Germany), an air-flow control unit (Akita(c), Activaero, Germany) and a custom-made sedimentation chamber. This chamber holds three Snapwell(c) inserts with monolayers of pulmonary epithelial cells. The whole set-up, referred to as the Pharmaceutical Aerosol Deposition Device On Cell Cultures (PADDOCC) system, aims to mimic the complete process of aerosol drug delivery, encompassing aerosol generation, aerosol deposition onto pulmonary epithelial cells and subsequent drug transport across this biological barrier, to facilitate the investigation of new aerosol formulations in the early stages of development. We describe here, the development of the design and the protocol for this device. By testing aerosol formulations of budesonide and salbutamol sulphate, respectively, reproducible deposition of aerosol particles on, and the integrity of, the pulmonary cell monolayer could be demonstrated. PMID:20822321

  16. Bilingual effects on deployment of the attention system in linguistically and culturally homogeneous children and adults.

    PubMed

    Yang, Sujin; Yang, Hwajin

    2016-06-01

    We investigated the impact of early childhood and adulthood bilingualism on the attention system in a group of linguistically and culturally homogeneous children (5- and 6-year olds) and young adults. We administered the child Attention Network Test (ANT) to 63 English monolingual and Korean-English bilingual children and administered the adult ANT to 39 language- and culture-matched college students. Advantageous bilingual effects on attention were observed for both children and adults in global processing levels of inverse efficiency, response time, and accuracy at a magnitude more pronounced for children than for adults. Differential bilingualism effects were evident at the local network level of executive control and orienting in favor of the adult bilinguals only. Notably, however, bilingual children achieved an adult level of accuracy in the incongruent flanker condition, implying enhanced attentional skills to cope with interferences. Our findings suggest that although both child and adult bilinguals share cognitive advantages in attentional functioning, age-related cognitive and linguistic maturation differentially shapes the outcomes of attentional processing at a local network level. PMID:26930166

  17. Cell Co-culture Patterning Using Aqueous Two-phase Systems

    PubMed Central

    Frampton, John P.; White, Joshua B.; Abraham, Abin T.; Takayama, Shuichi

    2013-01-01

    Cell patterning technologies that are fast, easy to use and affordable will be required for the future development of high throughput cell assays, platforms for studying cell-cell interactions and tissue engineered systems. This detailed protocol describes a method for generating co-cultures of cells using biocompatible solutions of dextran (DEX) and polyethylene glycol (PEG) that phase-separate when combined above threshold concentrations. Cells can be patterned in a variety of configurations using this method. Cell exclusion patterning can be performed by printing droplets of DEX on a substrate and covering them with a solution of PEG containing cells. The interfacial tension formed between the two polymer solutions causes cells to fall around the outside of the DEX droplet and form a circular clearing that can be used for migration assays. Cell islands can be patterned by dispensing a cell-rich DEX phase into a PEG solution or by covering the DEX droplet with a solution of PEG. Co-cultures can be formed directly by combining cell exclusion with DEX island patterning. These methods are compatible with a variety of liquid handling approaches, including manual micropipetting, and can be used with virtually any adherent cell type. PMID:23567187

  18. Real-time optical pH measurement in a standard microfluidic cell culture system.

    PubMed

    Magnusson, Einar B; Halldorsson, Skarphedinn; Fleming, Ronan M T; Leosson, Kristjan

    2013-01-01

    The rapid growth of microfluidic cell culturing in biological and biomedical research and industry calls for fast, non-invasive and reliable methods of evaluating conditions such as pH inside a microfluidic system. We show that by careful calibration it is possible to measure pH within microfluidic chambers with high accuracy and precision, using a direct single-pass measurement of light absorption in a commercially available phenol-red-containing cell culture medium. The measurement is carried out using a standard laboratory microscope and, contrary to previously reported methods, requires no modification of the microfluidic device design. We demonstrate the validity of this method by measuring absorption of light transmitted through 30-micrometer thick microfluidic chambers, using an inverted microscope fitted with a scientific-grade digital camera and two bandpass filters. In the pH range of 7-8, our measurements have a standard deviation and absolute error below 0.05 for a measurement volume smaller than 4 nL. PMID:24049695

  19. The Three-Dimensional Culture System with Matrigel and Neurotrophic Factors Preserves the Structure and Function of Spiral Ganglion Neuron In Vitro

    PubMed Central

    Sun, Gaoying; Liu, Wenwen; Fan, Zhaomin; Zhang, Daogong; Han, Yuechen; Xu, Lei; Qi, Jieyu; Zhang, Shasha; Gao, Bradley T.; Bai, Xiaohui; Li, Jianfeng; Chai, Renjie; Wang, Haibo

    2016-01-01

    Whole organ culture of the spiral ganglion region is a resourceful model system facilitating manipulation and analysis of live sprial ganglion neurons (SGNs). Three-dimensional (3D) cultures have been demonstrated to have many biomedical applications, but the effect of 3D culture in maintaining the SGNs structure and function in explant culture remains uninvestigated. In this study, we used the matrigel to encapsulate the spiral ganglion region isolated from neonatal mice. First, we optimized the matrigel concentration for the 3D culture system and found the 3D culture system protected the SGNs against apoptosis, preserved the structure of spiral ganglion region, and promoted the sprouting and outgrowth of SGNs neurites. Next, we found the 3D culture system promoted growth cone growth as evidenced by a higher average number and a longer average length of filopodia and a larger growth cone area. 3D culture system also significantly elevated the synapse density of SGNs. Last, we found that the 3D culture system combined with neurotrophic factors had accumulated effects in promoting the neurites outgrowth compared with 3D culture or NFs treatment only groups. Together, we conclude that the 3D culture system preserves the structure and function of SGN in explant culture. PMID:27057364

  20. A primary neuron culture system for the study of herpes simplex virus latency and reactivation.

    PubMed

    Kobayashi, Mariko; Kim, Ju-Youn; Camarena, Vladimir; Roehm, Pamela C; Chao, Moses V; Wilson, Angus C; Mohr, Ian

    2012-01-01

    Herpes simplex virus type-1 (HSV-1) establishes a life-long latent infection in peripheral neurons. This latent reservoir is the source of recurrent reactivation events that ensure transmission and contribute to clinical disease. Current antivirals do not impact the latent reservoir and there are no vaccines. While the molecular details of lytic replication are well-characterized, mechanisms controlling latency in neurons remain elusive. Our present understanding of latency is derived from in vivo studies using small animal models, which have been indispensable for defining viral gene requirements and the role of immune responses. However, it is impossible to distinguish specific effects on the virus-neuron relationship from more general consequences of infection mediated by immune or non-neuronal support cells in live animals. In addition, animal experimentation is costly, time-consuming, and limited in terms of available options for manipulating host processes. To overcome these limitations, a neuron-only system is desperately needed that reproduces the in vivo characteristics of latency and reactivation but offers the benefits of tissue culture in terms of homogeneity and accessibility. Here we present an in vitro model utilizing cultured primary sympathetic neurons from rat superior cervical ganglia (SCG) (Figure 1) to study HSV-1 latency and reactivation that fits most if not all of the desired criteria. After eliminating non-neuronal cells, near-homogeneous TrkA(+) neuron cultures are infected with HSV-1 in the presence of acyclovir (ACV) to suppress lytic replication. Following ACV removal, non-productive HSV-1 infections that faithfully exhibit accepted hallmarks of latency are efficiently established. Notably, lytic mRNAs, proteins, and infectious virus become undetectable, even in the absence of selection, but latency-associated transcript (LAT) expression persists in neuronal nuclei. Viral genomes are maintained at an average copy number of 25 per neuron

  1. A Primary Neuron Culture System for the Study of Herpes Simplex Virus Latency and Reactivation

    PubMed Central

    Kobayashi, Mariko; Kim, Ju-Youn; Camarena, Vladimir; Roehm, Pamela C.; Chao, Moses V.; Wilson, Angus C.; Mohr, Ian

    2012-01-01

    Herpes simplex virus type-1 (HSV-1) establishes a life-long latent infection in peripheral neurons. This latent reservoir is the source of recurrent reactivation events that ensure transmission and contribute to clinical disease. Current antivirals do not impact the latent reservoir and there are no vaccines. While the molecular details of lytic replication are well-characterized, mechanisms controlling latency in neurons remain elusive. Our present understanding of latency is derived from in vivo studies using small animal models, which have been indispensable for defining viral gene requirements and the role of immune responses. However, it is impossible to distinguish specific effects on the virus-neuron relationship from more general consequences of infection mediated by immune or non-neuronal support cells in live animals. In addition, animal experimentation is costly, time-consuming, and limited in terms of available options for manipulating host processes. To overcome these limitations, a neuron-only system is desperately needed that reproduces the in vivo characteristics of latency and reactivation but offers the benefits of tissue culture in terms of homogeneity and accessibility. Here we present an in vitro model utilizing cultured primary sympathetic neurons from rat superior cervical ganglia (SCG) (Figure 1) to study HSV-1 latency and reactivation that fits most if not all of the desired criteria. After eliminating non-neuronal cells, near-homogeneous TrkA+ neuron cultures are infected with HSV-1 in the presence of acyclovir (ACV) to suppress lytic replication. Following ACV removal, non-productive HSV-1 infections that faithfully exhibit accepted hallmarks of latency are efficiently established. Notably, lytic mRNAs, proteins, and infectious virus become undetectable, even in the absence of selection, but latency-associated transcript (LAT) expression persists in neuronal nuclei. Viral genomes are maintained at an average copy number of 25 per neuron

  2. Comparisons of multiple isotope systems in the aragonitic shells of cultured Arctica islandica clams

    NASA Astrophysics Data System (ADS)

    Liu, Y. W.; Aciego, S.; Wanamaker, A. D.

    2014-12-01

    Previous work using oxygen and stable carbon isotopes from Arctica islandica shells has shown that this archive can provide information on past seawater temperatures, carbon cycling and ocean circulation. However, relatively less attention has been devoted to other "non-traditional" isotope systems within this proxy archive. In this study, we report the boron (δ11B) and strontium isotopic values (87Sr/86Sr and δ88/86Sr) from A. islandicashells collected and cultured from the Gulf of Maine. The long-lived ocean quahog, A. islandica was collected and cultured in the Gulf of Maine for 8 months. Our high-resolution δ11B records from the experiment show 5-7‰ of increase through the culture, with low values from January to May and higher values after May. The 87Sr/86Sr ratios from both tank water and shell samples suggest that the shell material reflects ambient ocean chemistry without interferences from terrestrial sources. Although It has been suggested that stable Sr isotopic ratios (δ88/86Sr) in biogenic carbonates are influenced by the temperature of the precipitating fluid, our nearly identical δ88/86Sr data do not support this hypothesis despite a 15 °C temperature change during the experiment. Based on the in-situ measurements of culture seawater temperature, salinity and pH, and two commonly used fractionation factors (α3-4) for corals and forams, we predicted the range in shell δ11B values for the experiment. Our boron results are at the extreme ends of the two prediction lines suggesting the potential usage of the bivalve shells as seawater pH indicator. However, the wider range in δ11B in this experiment than the predictions based on other carbonate organisms (only 2 to 3‰) suggests that a species-specific fractionation factor may be required. Recent work from an additional constant temperature experiment (10 and 15 °C) in the Gulf of Maine will allow us to further evaluate temperature influences and potential vital effects on the shell boron

  3. Human retinal development in an in situ whole eye culture system.

    PubMed

    Engelsberg, Karl; Ghosh, Fredrik

    2011-01-01

    Phenotypic characterization of human retinogenesis may be facilitated by use of the tissue culture system paradigm. Traditionally, most culture protocols involve isolation of retinal tissue and/or cells, imposing various degrees of trauma, which in many cases leads to abnormal development. In this paper, we present a novel culture technique using whole embryonic eyes to investigate whether the retina in situ can develop normally in vitro. All procedures were carried out in accordance with the Declaration of Helsinki. Human embryos were obtained from elective abortions with the informed consent of the women seeking abortion. A total of 19 eyes were enucleated. The ages of the embryonic retinas were 6-7.5 weeks. Eyecups from 2 eyes were fixed immediately, to be used as controls. The remaining 17 eyes were placed on culture plates and divided into 3 groups kept for 7 (n = 4), 14 (n = 7) and 28 (n = 6) days in vitro (DIV). After fixation, the specimens were processed for hematoxylin and eosin staining, immunohistochemistry and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). Antibodies against recoverin (rods and cones), protein kinase C (PKC; rod bipolar cells) and vimentin (Müller cells) were used. TUNEL was used to detect apoptotic cells. In hematoxylin- and eosin-stained sections, the control retinas displayed a neuroblast cell layer (NBL) and an inner marginal zone. Specimens kept 7-14 DIV had a similar appearance, while 28-day specimens consisted of an NBL with almost no marginal zone. Thirteen of the 17 cultured retinas displayed completely normal lamination without rosettes or double folds. Pyknotic cells were found at the inner margin of the retinas, and the proportion of these cells increased with time in vitro. TUNEL staining revealed a few scattered cells in 7-DIV specimens, and the amount of stained cells in the inner part of the retinas progressively increased in 14- and 28-DIV specimens. Vimentin labeling showed cells

  4. Yield and gas exchange ability of sweetpotato plants cultured in a hydroponic system

    NASA Astrophysics Data System (ADS)

    Kitaya, Y.; Hirai, H.; Saiful Islam, A. F. M.; Yamamoto, M.

    Life support of crews in space is greatly dependent on the amounts of food atmospheric O 2 and clean water produced by plants Therefore the space farming systems with scheduling of crop production obtaining high yields with a rapid turnover rate converting atmospheric CO 2 to O 2 and purifying water should be established with employing suitable plant species and varieties and precisely controlling environmental variables around plants grown at a high density in a limited space In this study three sweetpotato varieties were cultured in a newly developed hydroponic system and the yield the photosynthetic rate and the transpiration rate were compared on the earth as a fundamental study for establishing the space farming systems The varieties were Elegant summer Koukei 14 and Beniazuma The hydroponic system mainly consisted of water channels and rockwool boards A growing space for roots was made between the rockwool board and nutrient solution in the water channel Storage roots were developed on the lower surface of the rockwool plates Fresh weights of the storage roots were 1 6 1 2 and 0 6 kg plant for Koukei 14 Elegant summer and Beniazuma respectively grown for five months from June to October under the sun light in Osaka Japan Koukei 14 and Elegant summer produced greater total phytomass than Beniazuma There were positive correlations among the total phytomass the net photosynthetic rate and the transpiration rate Young stems and leaves as well as storage roots of Elegant summer are edible Therefore Elegant-summer

  5. A Systems-Level Approach to Building Sustainable Assessment Cultures: Moderation, Quality Task Design and Dependability of Judgement

    ERIC Educational Resources Information Center

    Colbert, Peta; Wyatt-Smith, Claire; Klenowski, Val

    2012-01-01

    This article considers the conditions that are necessary at system and local levels for teacher assessment to be valid, reliable and rigorous. With sustainable assessment cultures as a goal, the article examines how education systems can support local-level efforts for quality learning and dependable teacher assessment. This is achieved through…

  6. A COUPLED MICROSOMAL-ACTIVATING/EMBRYO CULTURE SYSTEM: TOXICITY OF REDUCED BETA-NICOTINAMIDE ADENINE DINUCLEOTIDE PHOSPHATE (NADPH)

    EPA Science Inventory

    An NADPH-dependent microsomal-activating system has been coupled to a rat embryo culture in vitro. No embryonic morphological abnormalities or decrease in final yolk sac or embryo DNA and protein contents occurred when 0.2 mM NADPH was used in this coupled system. In contrast, 1....

  7. Localization of beta-adrenergic receptors on differentiated cells of the central nervous system in culture.

    PubMed Central

    Ventimiglia, R; Greene, M I; Geller, H M

    1987-01-01

    Hypothalamic neurons and cortical protoplasmic (type 1) astrocytes in dissociated cultures of rat brain express binding sites for antibodies specific for epitopes related to the beta-adrenergic receptor. Undifferentiated glial progenitor cells of the rat optic nerve do not detectably bind these antibodies, but both of the progeny [oligodendroglia and process-bearing (type 2) astrocytes] express immunologically identified beta-adrenergic receptors. Levels of receptor expression are variable: not all cells of each type express receptors nor is expression uniform on a given cell. The data suggest that cells of the central nervous system express beta-adrenergic receptors but that levels of expression may be determined by the differentiated state of the cells. Images PMID:3037533

  8. Algae for controlled ecological life support system diet characterization of cyanobacteria 'spirulina' in batch cultures

    NASA Technical Reports Server (NTRS)

    Tadros, M. G.

    1990-01-01

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

  9. Production of acetate from carbon dioxide in bioelectrochemical systems based on autotrophic mixed culture.

    PubMed

    Su, Min; Jiang, Yong; Li, Daping

    2013-08-01

    Bioelectrochemical systems (BESs) have been suggested as a new technology for wastewater treatment while accomplishing energy and chemical generation. This study describes the performance of BESs based on mixed culture that are capable of reducing carbon dioxide to acetate. The cathode potential was a critical factor that affected the performance of the BESs. The rate of acetate production increased as the electrode potential became more negative, from 0.38 mM d(-1) (-900 mV vs. Ag/AgCl) to 2.35 mM d(-1) (-1,100 mV), while the electron recovery efficiency of carbon dioxide reduction to acetate increased from 53.6% to 89.5%. The microbial population was dominated by relatives of Acetobacterium woodii when a methanogenic inhibitor was added to the BESs initially. PMID:23727797

  10. Toxicological effects of cypermethrin to marine phytoplankton in a co-culture system under laboratory conditions.

    PubMed

    Wang, Zhao-Hui; Nie, Xiang-Ping; Yue, Wen-Jie

    2011-08-01

    The growth of three marine phytoplankton species Skeletonema costatum, Scrippsiella trochoidea and Chattonella marina and the response of the antioxidant defense system have been investigated on exposure to commercial cypermethrin for 96 h and 32 days in a co-culture system. Growth of the three species was generally comparable over 96 h with an inoculation of 1:3:6.5 (C. marina:S. trochoidea:S. costatum), with stimulation at 5 μg l(-1) and inhibition under higher concentrations (50, 100 μg l(-1)). However, when inoculating at ratios of 1:1:1 during a 32 day test, S. costatum became the most sensitive species and was significantly inhibited in all test groups under the dual stresses of cypermethrin and interspecies competition. The growth of C. marina was significantly inhibited at the concentrations higher than 5 μg l(-1), while the growth of S. trochoidea was significantly promoted at low concentrations. Superoxide dismutase (SOD) activities significantly increased during 6-12 h exposure periods in test treatments at low concentrations, and enhanced in the control as well due to interspecies competition. The lipid peroxidation product malondialdehyde was enhanced at high concentrations, but did not increase in control and low concentration cultures with high SOD activities, indicating that algal cells activated the antioxidant enzymes promptly to protect the cells from lipid membrane damage. Results from this study suggested that cypermethrin pollution in maricultural sea waters might lead to a shift in phytoplankton community structure from diatom to harmful dinoflagellate species, and thus potentially stimulatory for harmful algal blooms. PMID:21499869

  11. Culture-Independent Analysis of Aerosol Microbiology in a Metropolitan Subway System

    PubMed Central

    Robertson, Charles E.; Baumgartner, Laura K.; Harris, J. Kirk; Peterson, Kristen L.; Stevens, Mark J.; Frank, Daniel N.

    2013-01-01

    The goal of this study was to determine the composition and diversity of microorganisms associated with bioaerosols in a heavily trafficked metropolitan subway environment. We collected bioaerosols by fluid impingement on several New York City subway platforms and associated sites in three sampling sessions over a 1.5-year period. The types and quantities of aerosolized microorganisms were determined by culture-independent phylogenetic analysis of small-subunit rRNA gene sequences by using both Sanger (universal) and pyrosequencing (bacterial) technologies. Overall, the subway bacterial composition was relatively simple; only 26 taxonomic families made up ∼75% of the sequences determined. The microbiology was more or less similar throughout the system and with time and was most similar to outdoor air, consistent with highly efficient air mixing in the system. Identifiable bacterial sequences indicated that the subway aerosol assemblage was composed of a mixture of genera and species characteristic of soil, environmental water, and human skin commensal bacteria. Eukaryotic diversity was mainly fungal, dominated by organisms of types associated with wood rot. Human skin bacterial species (at 99% rRNA sequence identity) included the Staphylococcus spp. Staphylococcus epidermidis (the most abundant and prevalent commensal of the human integument), S. hominis, S. cohnii, S. caprae, and S. haemolyticus, all well-documented human commensal bacteria. We encountered no organisms of public health concern. This study is the most extensive culture-independent survey of subway microbiota so far and puts in place pre-event information required for any bioterrorism surveillance activities or monitoring of the microbiological impact of recent subway flooding events. PMID:23542619

  12. X-ray fluorescence imaging system for fast mapping of pigment distributions in cultural heritage paintings

    NASA Astrophysics Data System (ADS)

    Zielińska, A.; Dąbrowski, W.; Fiutowski, T.; Mindur, B.; Wiącek, P.; Wróbel, P.

    2013-10-01

    Conventional X-ray fluorescence imaging technique uses a focused X-ray beam to scan through the sample and an X-ray detector with high energy resolution but no spatial resolution. The spatial resolution of the image is then determined by the size of the exciting beam, which can be obtained either from a synchrotron source or from an X-ray tube with a micro-capillary lens. Such a technique based on a pixel-by-pixel measurement is very slow and not suitable for imaging large area samples. The goal of this work is to develop a system capable of simultaneous imaging of large area samples by using a wide field uniform excitation X-ray beam and a position sensitive and energy dispersive detector. The development is driven by possible application of such a system to imaging of distributions of hidden pigments containing specific elements in cultural heritage paintings, which is of great interest for the cultural heritage research. The fluorescence radiation from the area of 10 × 10 cm2 is projected through a pinhole camera on the Gas Electron Multiplier detector of the same area. The detector is equipped with two sets of orthogonal readout strips. The strips are read out by the GEMROC Application Specific Integrated Circuits (ASIC)s, which deliver time and amplitude information for each hit. This ASIC architecture combined with a Field Programmable Gate Array (FPGA) based readout system allows us to reconstruct the position and the total energy of each detected photon for high count rates up to 5 × 106 cps. Energy resolution better than 20% FWHM for the 5.9 keV line and spatial resolution of 1 mm FWHM have been achieved for the prototype system. Although the energy resolution of the Gas Electron Multiplier (GEM) detector is, by principle, not competitive with that of specialised high energy resolution semiconductor detectors, it is sufficient for a number of applications. Compared to conventional micro-XRF techniques the developed system allows shortening of the

  13. Gastric culture

    MedlinePlus

    ... test or procedure preparation (3 to 6 years) School age test or procedure preparation (6 to 12 ... immune system. The final results of the gastric culture test may take several weeks. Your provider will ...

  14. Closed System Cell Culture Protocol Using HYPERStack Vessels with Gas Permeable Material Technology

    PubMed Central

    Rothenberg, Mark; Martin, Greg

    2010-01-01

    Large volume adherent cell culture is currently standardized on stacked plate cell growth products when microcarrier beads are not an optimal choice. HYPERStack vessels allow closed system scale up from the current stacked plate products and delivers >2.5X more cells in the same volumetric footprint. The HYPERStack vessels function via gas permeable material which allows gas exchange to occur, therefore eliminating the need for internal headspace within a vessel. The elimination of headspace allows the compartment where cell growth occurs to be minimized to reduce space, allowing more layers of cell growth surface area within the same volumetric footprint. For many applications such as cell therapy or vaccine production, a closed system is required for cell growth and harvesting. The HYPERStack vessel allows cell and reagent addition and removal via tubing from media bags or other methods. This protocol will explain the technology behind the gas permeable material used in the HYPERStack vessels, gas diffusion results to meet the metabolic needs of cells, closed system cell growth protocols, and various harvesting methods. PMID:21189467

  15. A plant culture system for producing food and recycling materials with sweetpotato in space

    NASA Astrophysics Data System (ADS)

    Kitaya, Yoshiaki; Yano, Sachiko; Hirai, Hiroaki

    2016-07-01

    The long term human life support in space is greatly dependent on the amounts of food, atmospheric O2 and clean water produced by plants. Therefore, the bio-regenerative life support system such as space farming with scheduling of crop production, obtaining high yields with a rapid turnover rate, converting atmospheric CO2 to O2 and purifying water should be established with employing suitable plant species and varieties and precisely controlling environmental variables around plants grown at a high density in a limited space. We are developing a sweetpotato culture system for producing tuberous roots as a high-calorie food and fresh edible leaves and stems as a nutritive functional vegetable food in space. In this study, we investigated the ability of food production, CO2 to O2 conversion through photosynthesis, and clean water production through transpiration in the sweetpotato production system. The biomass of edible parts in the whole plant was almost 100%. The proportion of the top (leaves and stems) and tuberous roots was strongly affected by environmental variables even when the total biomass production was mostly the same. The production of biomass and clean water was controllable especially by light, atmospheric CO2 and moisture and gas regimes in the root zone. It was confirmed that sweetpotato can be utilized for the vegetable crop as well as the root crop allowing a little waste and is a promising functional crop for supporting long-duration human activity in space.

  16. Contamination and growth of the shrimp, Penaeus stylirostris Stimpson, cultured in a seawater/wastewater aquaculture system

    SciTech Connect

    Landau, M.; Pierce, R.H.; Williams, L.D.; Norris, D.R.

    1985-10-01

    Many commercial mariculture operations have failed because of the costs associated with rearing animals in high density cultures. One of the most significant costs is feed. The costs of culturing marine animals could be lowered by reducing the stocking densities used in the ponds and substituting nutrient-rich, treated wastewater for commercial feeds. However, an aquaculture system which is based on wastewater may not only have a different production profile than a system based on prepared feeds, but the animals may also be subject to contamination. The authors have monitored the growth of shrimp cultured in a seawater/wastewater system and have analyzed water and tissue samples for several likely organic and inorganic contaminants.

  17. Comparison of In Vitro- and Chorioallantoic Membrane (CAM)-Culture Systems for Cryopreserved Medulla-Contained Human Ovarian Tissue

    PubMed Central

    Isachenko, Vladimir; Mallmann, Peter; Petrunkina, Anna M.; Rahimi, Gohar; Nawroth, Frank; Hancke, Katharina; Felberbaum, Ricardo; Genze, Felicitas; Damjanoski, Ilija; Isachenko, Evgenia

    2012-01-01

    At present, there are three ways to determine effectively the quality of the cryopreservation procedure using ovarian tissue before the re-implantation treatment: evaluation of follicles after post-thawing xenotransplantation to SCID mouse, in-vitro culture in a large volume of culture medium under constant agitation and culture on embryonic chorio-allantoic membrane within a hen's eggs. The aim of this study was to compare the two methods, culture in vitro and culture on embryonic chorioallantoic membrane (CAM) of cryopreserved human ovarian medulla-contained and medulla-free cortex. Ovarian fragments were divided into small pieces (1.5–2.0×1.0–1.2×0.8–1.5) of two types, cortex with medulla and medulla-free cortex, frozen, thawed and randomly divided into the following four groups. Group 1: medulla-free cortex cultured in vitro for 8 days in large volume of medium with mechanical agitation, Group 2: medulla-containing cortex cultured in vitro, Group 3: medulla-free cortex cultured in CAM-system for 5 days, Group 4: medulla-containing cortex cultured in CAM-system. The efficacy of the tissue culture was evaluated by the development of follicles and by intensiveness of angiogenesis in the tissue (von Willebrand factor and Desmin). For Group 1, 2, 3 and 4, respectively 85%, 85%, 87% and 84% of the follicles were morphologically normal (P>0.1). The immunohistochemical analysis showed that angiogenesis detected by von Willebrand factor was lower in groups 1 and 3 (medulla-free cortex). Neo-vascularisation (by Desmin) was observed only in ovarian tissue of Group 4 (medulla-contained cortex after CAM-culture). It appears that the presence of medulla in ovarian pieces is beneficial for post-thaw development of cryopreserved human ovarian tissue. For medical practice it is recommended for evaluation of post-warming ovarian tissue to use the CAM-system as a valuable alternative to xenotransplantation and for cryopreservation of these tissues to prepare ovarian

  18. A personalized and control systems engineering conceptual approach to target childhood anxiety in the contexts of cultural diversity.

    PubMed

    Pina, Armando A; Holly, Lindsay E; Zerr, Argero A; Rivera, Daniel E

    2014-01-01

    In the child and adolescent anxiety area, some progress has been made to develop evidence-based prevention protocols, but less is known about how to best target these problems in children and families of color. In general, data show differential program effects with some minority children benefiting significantly less. Our preliminary data, however, show promise and suggest cultural parameters to consider in the tailoring process beyond language and cultural symbols. It appears that a more focused approach to culture might help activate intervention components and its intended effects by focusing, for example, on the various facets of familismo when working with some Mexican parents. However, testing the effects and nuances of cultural adaption vis-à-vis a focused personalized approach is methodologically challenging. For this reason, we identify control systems engineering design methods and provide example scenarios relevant to our data and recent intervention work. PMID:24702279

  19. A Personalized and Control Systems Engineering Conceptual Approach to Target Childhood Anxiety in the Contexts of Cultural Diversity

    PubMed Central

    Pina, Armando A.; Holly, Lindsay E.; Zerr, Argero A.; Rivera, Daniel E.

    2014-01-01

    In the child and adolescent anxiety area, some progress has been made to develop evidence-based prevention protocols but less is known about how to best target these problems in children and families of color. In general, data show differential program effects with some minority children benefiting significantly less. Our preliminary data, however, show promise and suggest cultural parameters to consider in the tailoring process beyond language and cultural symbols. It appears that a more focused approach to culture might help activate intervention components and its intended effects by focusing, for example, on the various facets of familismo when working with some Mexican parents. However, testing the effects and nuances of cultural adaption vis-à-vis a focused personalized approach is methodologically challenging. For this reason, we identify control systems engineering design methods and provide example scenarios relevant to our data and recent intervention work. PMID:24702279

  20. Expression of amino acid transport systems in cultured human umbilical vein endothelial cells.

    PubMed

    Mann, G E; Pearson, J D; Sheriff, C J; Toothill, V J

    1989-03-01

    1. Nutrient transport in cultured human umbilical vein endothelial cells was characterized using a rapid dual-isotope dilution technique. Microcarrier beads with confluent endothelial cells were perfused in small columns, and uptake and efflux were assessed relative to D-mannitol (extracellular tracer) during a single transit through the column. 2. At tracer concentrations significant unidirectional uptakes were measured for L-leucine (53 +/- 2%), L-phenylalanine (73 +/- 2%), L-serine (40 +/- 4%), L-arginine (42 +/- 3%) and L-ornithine (26 +/- 3%). Uptake for L-proline, D-glucose, dopamine and serotonin was lower (6-10%), whereas uptake for the system A analogue 2-methylaminoisobutyric acid (2-MeAIB) was negligible. Uptakes rapidly decreased with time due to tracer efflux. 3. Endothelial cell transport of L-leucine was markedly inhibited during perfusion with 1 mM-BCH (beta-2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid, system L analogue), L-leucine, D-leucine, L-phenylalanine, L-methionine and L-DOPA. 2-MeAIB, L-cysteine, glycine, L-proline, hydroxy-L-proline, L-aspartate and beta-alanine were poor inhibitors, while L-serine and the cationic substrates L-lysine and L-arginine inhibited uptake by 10-35%. 4. When the kinetics of L-leucine transport were examined over a wide range of substrate concentrations (0.025-1 mM) transport was saturable. A single entry site analysis gave a half-maximal saturation constant Kt = 0.24 +/- 0.08 mM (mean +/- S.E.M., n = 5) and a Vmax = 27.8 +/- 4.6 nmol/min per column (approximately 3 x 10(6) cells). 5. Removal of sodium from the perfusate inhibited tracer uptake of L-leucine, L-serine and L-arginine by respectively 20 +/- 5% (n = 3), 77 +/- 5% (n = 3) and 35 +/- 4% (n = 3). 6. Our results provide the first evidence that cultured human endothelial cells of venous origin express a saturable transport system for large neutral amino acids resembling system L described in brain microvascular endothelium. Detection of Na

  1. Chlorella vulgaris culture as a regulator of CO2 in a bioregenerative life support system

    NASA Astrophysics Data System (ADS)

    Li, Ming; Hu, Dawei; Liu, Hong; Hu, Enzhu; Xie, Beizhen; Tong, Ling

    2013-08-01

    It is the primary task for a bioregenerative life support system (BLSS) to maintain the stable concentrations of CO2 and O2. However, these concentrations could fluctuate based on various factors, such as the imbalance between respiration/assimilation quotients of the heterotrophic and autotrophic components. They can even be out of balance through catastrophic failure of higher plants in the emergency conditions. In this study, the feasibility of using unicellular Chlorella vulgaris of typically rapid growth as both “compensatory system” and “regulator” to control the balance of CO2 and O2 was analyzed in a closed ecosystem. For this purpose, a small closed ecosystem called integrative experimental system (IES) was established in our laboratory where we have been conducting multi-biological life support system experiments (MLSSE). The IES consists of a closed integrative cultivating system (CICS) and a plate photo-bioreactor. Four volunteers participated in the study for gas exchange by periodical breathing through a tube connected with the CICS. The plate photo-bioreactor was used to cultivate C. vulgaris. Results showed that the culture of C. vulgaris could be used in a situation of catastrophic failure of higher plant under the emergencies. And the productivity could recover itself to the original state in 3 to 5 days to protect the system till the higher plant was renewed. Besides, C. vulgaris could grow well and the productivity could be affected by the light intensity which could help to keep the balance of CO2 and O2 in the IES efficiently. Thus, C. vulgaris could be included in the design of a BLSS as a “compensatory system” in the emergency contingency and a “regulator” during the normal maintenance.

  2. Biological characteristics of adult degenerative nucleus pulposus cells in a three-dimensional microcarrier stirring culture system.

    PubMed

    Ning, Bin; Liu, Haifei; Gong, Weiming; Jiang, Jianhao; Hu, Yougu; Yang, Shang-You

    2013-06-01

    A major problem in reconstructing degenerative intervertebral discs is to obtain sufficient nucleus pulposus (NP) seeding cells with normal physiologic functions. The current study adopted a three-dimensional microcarrier culture system for massive cell expansion and evaluated the biological characteristics and physiological functions of the propagated adult degenerative NP cells. Isolated adult NP cells were cultured in either microcarrier stirring culturing system or traditional monolayer cultivation. The growth characteristics, proliferation, extracellular matrix secretion, and apoptosis potential were examined to evaluate the different features of the two cultivation methods. Compared to the monolayer cultivation system, the adhesion time of NP cells in the three-dimensional microcarrier culture system appeared longer with relatively transient stable growth period. MTT and (3)H-TdR assays suggested significantly elevated proliferation and higher thymidine incorporation rates in cells from microcarrier system compare to cells in the monolayer system at the exponential growth phase (p < 0.05). Western blot data complimented the immunostaining results that the NP cells in the microcarrier system expressed significantly more protein levels of both type collagens at the exponential growth phase than that in the monolayer system (p < 0.05). Further, significantly more (35)S labeled proteoglycan incorporation was noticed in the cells on the microcarriers at both the stable growth and the exponential growth phases (p < 0.05 and p < 0.01). In conclusion, the three-dimensional microcarrier stirring culture system provides a means of fast and massive propagation of NP seeding cells which maintain their normal physiological characteristics and functions. PMID:23335154

  3. Portable self-contained cultures for phage and bacteria made of paper and tape.

    PubMed

    Funes-Huacca, Maribel; Wu, Alyson; Szepesvari, Eszter; Rajendran, Pavithra; Kwan-Wong, Nicholas; Razgulin, Andrew; Shen, Yi; Kagira, John; Campbell, Robert; Derda, Ratmir

    2012-11-01

    In this paper, we demonstrate that a functional, portable device for the growth of bacteria or amplification of bacteriophage can be created using simple materials. These devices are comprised of packing tape, sheets of paper patterned by hydrophobic printer ink, and a polydimethyl siloxane (PDMS) membrane, which is selectively permeable to oxygen but non-permeable to water. These devices supply bacteria with oxygen and prevent the evaporation of media for a period over 48 h. The division time of E. coli and the amplification of the phage M13 in this device are similar to the rates measured on agar plates and in shaking cultures. The growth of bacteria with a fluorescent mCherry reporter can be quantified using a flatbed scanner or a cell phone camera. Permeating devices with commercial viability dye (PrestoBlue) can be used to detect low copy number of E. coli (1-10 CFU in 100 μL) and visualize microorganisms in environmental samples. The platform, equipped with bacteria that carry inducible mCherry reporter could also be used to quantify the concentration of the inducer (here, arabinose). Identical culture platforms can, potentially, be used to quantify the induction of gene expression by an engineered phage or by synthetic transcriptional regulators that respond to clinically relevant molecules. The majority of measurement and fabrication procedures presented in this report have been replicated by low-skilled personnel (high-school students) in a low-resource environment (high-school classroom). The fabrication and performance of the device have also been tested in a low-resource laboratory setting by researchers in Nairobi, Kenya. Accordingly, this platform can be used as both an educational tool and as a diagnostic tool in low-resource environments worldwide. PMID:22895550

  4. Characterization of an in vitro cell culture bioreactor system to evaluate anti-neoplastic drug regimens.

    PubMed

    Kirstein, Mark N; Brundage, Richard C; Elmquist, William F; Remmel, Rory P; Marker, Paul H; Guire, Dan E; Yee, Douglas

    2006-04-01

    A dynamic 3-dimensional tissue culture system has been developed that will allow for control of gemcitabine exposure to mimic concentration-time profiles measured from biologic samples. Gemcitabine was infused into a central reservoir. Media is mixed and delivered through hollow fiber capillaries, where it diffuses into the extracapillary space containing anchorage-dependent MDA-231 cells. To test for control of gemcitabine concentration-time profiles, drug was first infused through bioreactors without cells, and gemcitabine concentrations were measured with HPLC. Concentrations could be controlled to simulate 30-min and 2.5 h infusions, and were similar in both the lumen and extracapillary space. MDA-231 cells were then seeded into control (n = 4) and gemcitabine treatment (n = 4) groups, and maintained in culture for 2 weeks. Gemcitabine (5.3 mg) was infused over 30 min to the treatment group, and blank media to the control group. Accuracy of measured gemcitabine maximum concentration (Cmax) was 83.4%, and area under the curve (AUC), 106.2%, relative to pre-experimental theoretical values. With cells present, gemcitabine AUC in the extracapillary space was 32% of the value in the lumen. For the control group, 21.2 million cells (94.3% viable) were recovered, and for the gemcitabine-treated group, 16.8 million cells (87.1 % viable). Flow cytometry showed that 13.3 % of cells in the control group were in S-phase and 34.3 % in the gemcitabine-treated group were in S-phase (p = 0.003). In conclusion, gemcitabine concentration-time profiles could be accurately controlled through dosage, infusion rate, and pump flow rate, and cells could be recovered afterward to evaluate drug treatment. PMID:16502018

  5. Ho:YAG laser: intervertebral disk cell interaction using three-dimensional cell culture system

    NASA Astrophysics Data System (ADS)

    Sato, Masato; Ishihara, Miya; Arai, Tsunenori; Asazuma, Takashi; Kikuchi, Toshiyuki; Kikuchi, Makoto; Fujikawa, Kyosuke

    2000-06-01

    The purpose of this study is to evaluate the influence on the intervertebral disc cells after laser irradiation using three- dimensional culture system and to clarify the optimum Ho:YAG laser irradiation condition on percutaneous laser disc decompression (PLDD) for lumbar disc herniation. Since the Ho:YAG laser ablation is characterized by water-vapor bubble dynamics, not only thermal effect but also acoustic effect on cell metabolism might occur in the intervertebral disc. We studied the disc cell reaction from the metabolic point of view to investigate photothermal and photoacoustic effects on three-dimensional cultured disc cell. Intervertebral discs were obtained from female 30 Japanese white rabbits weighing about 1 kg. A pulsed Ho:YAG laser (wavelength: 2.1 micrometer, pulse width: about 200 microseconds) was delivered through a 200 micrometer-core diameter single silica glass fiber. We used the Ho:YAG laser irradiation fluence ranging from 60 to approximately 800 J/cm2 at the fiber end. To investigate acoustic effect, the acoustic transducer constructed with polyvinylidene fluoride (PVdF) film and acoustic absorber was used to detect the stress wave. Thermocouple and thermography were used to investigate thermal effect. Concerning damage of plasma membrane and ability of matrix synthesis, thermal effect might mainly affect cell reaction in total energy of 54 J (closed to practically used condition), but in 27 J, acoustic effect might contribute to it. We found that total energy was key parameter among the optimum condition, so that temperature and/or stress wave may influence Ho:YAG laser-disc cell interactions.

  6. The impact of caffeine on connexin expression in the embryonic chick cardiomyocyte micromass culture system.

    PubMed

    Ahir, Bhavesh K; Pratten, Margaret K

    2016-07-01

    Cardiomyocytes are electrically coupled by gap junctions, defined as clusters of low-resistance multisubunit transmembrane channels composed of connexins (Cxs). The expression of Cx40, Cx43 and Cx45, which are present in cardiomyocytes, is known to be developmentally regulated. This study investigates the premise that alterations in gap junction proteins are one of the mechanisms by which teratogens may act. Specifically, those molecules known to be teratogenic in humans could cause their effects via disruption of cell-to-cell communication pathways, resulting in an inability to co-ordinate tissue development. Caffeine significantly inhibited contractile activity at concentrations above and including 1500 μm (P < 0.05), while not affecting cell viability and total protein, in the embryonic chick cardiomyocyte micromass culture system. The effects of caffeine on key cardiac gap junction protein (Cx40, Cx43 and Cx45) expression were analysed using immunocytochemistry and in-cell Western blotting. The results indicated that caffeine altered the expression pattern of Cx40, Cx43 and Cx45 at non-cytotoxic concentrations (≥2000 μm), i.e., at concentrations that did not affect total cell protein and cell viability. In addition the effects of caffeine on cardiomyocyte formation and function (contractile activity score) were correlated with modulation of Cxs (Cx40, Cx43 and Cx45) expression, at above and including 2000 μm caffeine concentrations (P < 0.05). These experiments provide evidence that embryonic chick cardiomyocyte micromass culture may be a useful in vitro method for mechanistic studies of perturbation of embryonic heart development. Copyright © 2015 John Wiley & Sons, Ltd. PMID:26304238

  7. Cytotoxic effects of gold nanoparticles exposure employing in vitro animal cell culture system as part of nanobiosafety

    NASA Astrophysics Data System (ADS)

    Ambwani, Sonu; Kakade Datta, P.; Kandpal, Deepika; Arora, Sandeep; Ambwani, Tanuj Kumar

    2016-04-01

    Metal Nanoparticles are exploited in different fields that include biomedical sector where they are utilized in drug and gene delivery, biosensors, cancer treatment and diagnostic tools. Despite of their benefits, there has been serious concerns about possible side effects of several nanoparticles. Gold nanoparticles (AuNPs) are exploited for bio-imaging, biosensing, drug delivery, transfection and diagnosis. These nanoparticles may get released into the environment in high amounts at all stages of production, recycling and disposal. Since the manufacture and use of nanoparticles are increasing, humans/ animals are more likely to be exposed occupationally or via consumer products and the environment. The emergence of the new field of nanotoxicity has spurred great interest in a wide variety of materials and their possible effects on living systems. Animal cell culture system is considered as a sensitive indicator against exposure of such materials. Keeping in view the above scenario, present study was carried out to evaluate effect of AuNPs exposure in primary and cell line culture system employing chicken embryo fibroblast (CEF) culture and HeLa cell line culture through MTT assay. Minimum cytotoxic dose was found to be 60 µg/ml and 50 µg/ml in CEF and HeLa cells, respectively. Thus, it could be inferred that even a very low concentration of AuNPs could lead to cytotoxic effects in cell culture based studies.

  8. Characterization of a rat in vitro ovarian culture system to study the ovarian toxicant 4-vinylcyclohexene diepoxide.

    PubMed

    Devine, Patrick J; Sipes, I Glenn; Skinner, Michael K; Hoyer, Patricia B

    2002-10-15

    Repeated daily dosing of rats with the occupational chemical 4-vinylcyclohexene diepoxide (VCD) causes selective depletion of the smallest preantral ovarian follicles (primordial and primary). These targeted populations are difficult to study because they comprise very little of the overall mass of ovarian tissue. Additionally, they are randomly distributed throughout the ovary. Therefore, a neonatal rat ovarian culture system containing predominantly primordial and primary follicles was developed and its susceptibility to VCD was assessed. The in vivo sensitivity of neonatal rats to VCD dosing was first confirmed by daily injection of VCD (80 mg/kg/day ip) on postnatal days (PND) 4-19. On PND 19, depletion of primordial and small primary follicles was evident. Ovarian cultures were then established utilizing a floating organ culture system to treat ovaries from PND 4 Fischer 344 rats in vitro. Initial follicle populations in cultured ovaries consisted of primordial (81%) and small primary (19%) follicles, whereas larger-sized preantral follicles had developed after 15 days in culture (67% primordial, 31% small primary, and 2% large primary). Cultured rat ovaries were sensitive to follicle depletion by incubation with VCD (> or =30 microM), and follicle loss occurred in a time-dependent manner (8-15 days). Evidence for apoptosis in VCD-exposed ovaries, as demonstrated in vivo, was obtained using immunohistochemistry. There was significantly more staining for apoptosis-associated active caspase-3 and TUNEL in ovaries incubated with VCD (30 microM, 15 days) compared with matched controls. These results demonstrate that small preantral follicles of cultured neonatal rat ovaries are sensitive to VCD exposure. The similarities between VCD's effects in vitro and in vivo demonstrate the usefulness of this system for future mechanistic studies related to ovarian follicle loss induced by VCD or other ovotoxic chemicals. PMID:12408955

  9. A screen-printed microband glucose biosensor system for real-time monitoring of toxicity in cell culture.

    PubMed

    Pemberton, R M; Xu, J; Pittson, R; Drago, G A; Griffiths, J; Jackson, S K; Hart, J P

    2011-01-15

    Microband biosensors, screen-printed from a water-based carbon ink containing cobalt phthalocyanine redox mediator and glucose oxidase (GOD) enzyme, were used to monitor glucose levels continuously in buffer and culture medium. Five biosensors were operated amperometrically (E(app) of +0.4V), in a 12-well tissue culture plate system at 37°C, using a multipotentiostat. After 24 h, a linear calibration plot was obtained from steady-state current responses for glucose concentrations up to 10 mM (dynamic range 30 mM). Within the linear region, a correlation coefficient (R(2)) of 0.981 was obtained between biosensor and spectrophotometric assays. Over 24 h, an estimated 0.15% (89 nmol) of the starting glucose concentration (24 mM) was consumed by the microbiosensor. The sensitivity of the biosensor response in full culture medium was stable between pHs 7.3 and 8.4. Amperometric responses for HepG2 monolayer cultures decreased with time in inverse proportionality to cell number (for 0 to 10(6) cell/ml), as glucose was being metabolised. HepG2 3D cultures (spheroids) were also shown to metabolise glucose, at a rate which was independent of spheroid age (between 6 and 15 days). Spheroids were used to assay the effect of a typical hepatotoxin, paracetamol. At 1 mM paracetamol, glucose uptake was inhibited by 95% after 6 h in culture; at 500 μM, around 15% inhibition was observed after 16 h. This microband biosensor culture system could form the basis for an in vitro toxicity testing system. PMID:21081270

  10. Development of an Insert Co-culture System of Two Cellular Types in the Absence of Cell-Cell Contact.

    PubMed

    Renaud, Justine; Martinoli, Maria-Grazia

    2016-01-01

    The role of secreted soluble factors in the modification of cellular responses is a recurrent theme in the study of all tissues and systems. In an attempt to make straightforward the very complex relationships between the several cellular subtypes that compose multicellular organisms, in vitro techniques have been developed to help researchers acquire a detailed understanding of single cell populations. One of these techniques uses inserts with a permeable membrane allowing secreted soluble factors to diffuse. Thus, a population of cells grown in inserts can be co-cultured in a well or dish containing a different cell type for evaluating cellular changes following paracrine signaling in the absence of cell-cell contact. Such insert co-culture systems offer various advantages over other co-culture techniques, namely bidirectional signaling, conserved cell polarity and population-specific detection of cellular changes. In addition to being utilized in the field of inflammation, cancer, angiogenesis and differentiation, these co-culture systems are of prime importance in the study of the intricate relationships that exist between the different cellular subtypes present in the central nervous system, particularly in the context of neuroinflammation. This article offers general methodological guidelines in order to set up an experiment in order to evaluating cellular changes mediated by secreted soluble factors using an insert co-culture system. Moreover, a specific protocol to measure the neuroinflammatory effects of cytokines secreted by lipopolysaccharide-activated N9 microglia on neuronal PC12 cells will be detailed, offering a concrete understanding of insert co-culture methodology. PMID:27500972

  11. Evaluation of Mouse Oocyte In Vitro Maturation Developmental Competency in Dynamic Culture Systems by Design and Construction of A Lab on A Chip Device and Its Comparison with Conventional Culture System

    PubMed Central

    Sadeghzadeh Oskouei, Behnaz; Pashaiasl, Maryam; Heidari, Mohammad Hasan; Salehi, Mohammad; Veladi, Hadi; Ghaderi Pakdel, Firuz; Shahabi, Parviz; Novin, Marefat Ghaffari

    2016-01-01

    Objective In conventional assisted reproductive technology (ART), oocytes are cultured in static microdrops within Petri dishes that contain vast amounts of media. However, the in vivo environment is dynamic. This study assesses in vitro oocyte maturation through the use of a new microfluidic device. We evaluate oocyte fertilization to the blastocyct stage and their glutathione (GSH) contents in each experimental group. Materials and Methods In this experimental study, we established a dynamic culture condition. Immature oocytes were harvested from ovaries of Naval Medical Research Institute (NMRI) mice. Oocytes were randomly placed in static (passive) and dynamic (active) in vitro maturation (IVM) culture medium for 24 hours. In vitro matured oocytes underwent fertilization, after which we placed the pronucleus (PN) stage embryos in microdrops and followed their developmental stages to blastocyst formation after 3 days. GSH content of the in vitro matured oocytes was assessed by monochlorobimane (MCB) staining. Results We observed significantly higher percentages of mature metaphase II oocytes (MII) in the passive and active dynamic culture systems (DCS) compared to the static group (P<0.01). There were significantly less mean numbers of germinal vesicle (GV) and degenerated oocytes in the passive and active dynamic groups compared to the static group (P<0.01). Fertilization and blastocyst formation rate in the dynamic systems were statistically significant compared to the static cultures (P<0.01). There was significantly higher GSH content in dynamically matured oocytes compared to statically matured oocytes (P<0.01). Conclusion Dynamic culture for in vitro oocyte maturation improves their developmental competency in comparison with static culture conditions. PMID:27540525

  12. A biofidelic 3D culture model to study the development of brain cellular systems.

    PubMed

    Ren, M; Du, C; Herrero Acero, E; Tang-Schomer, M D; Özkucur, N

    2016-01-01

    Little is known about how cells assemble as systems during corticogenesis to generate collective functions. We built a neurobiology platform that consists of fetal rat cerebral cortical cells grown within 3D silk scaffolds (SF). Ivermectin (Ivm), a glycine receptor (GLR) agonist, was used to modulate cell resting membrane potential (Vmem) according to methods described in a previous work that implicated Ivm in the arrangement and connectivity of cortical cell assemblies. The cells developed into distinct populations of neuroglial stem/progenitor cells, mature neurons or epithelial-mesenchymal cells. Importantly, the synchronized electrical activity in the newly developed cortical assemblies could be recorded as local field potential (LFP) measurements. This study therefore describes the first example of the development of a biologically relevant cortical plate assembly outside of the body. This model provides i) a preclinical basis for engineering cerebral cortex tissue autografts and ii) a biofidelic 3D culture model for investigating biologically relevant processes during the functional development of cerebral cortical cellular systems. PMID:27112667

  13. Monument Damage Information System (mondis): AN Ontological Approach to Cultural Heritage Documentation

    NASA Astrophysics Data System (ADS)

    Cacciotti, R.; Valach, J.; Kuneš, P.; Čerňanský, M.; Blaško, M.; Křemen, P.

    2013-07-01

    Deriving from the complex nature of cultural heritage conservation it is the need for enhancing a systematic but flexible organization of expert knowledge in the field. Such organization should address comprehensively the interrelations and complementariness among the different factors that come into play in the understanding of diagnostic and intervention problems. The purpose of MONDIS is to endorse this kind of organization. The approach consists in applying an ontological representation to the field of heritage conservation in order to establish an appropriate processing of data. The system allows replicating in a computer readable form the basic dependence among factors influencing the description, diagnosis and intervention of damages to immovable objects. More specifically MONDIS allows to input and search entries concerning object description, structural evolution, location characteristics and risk, component, material properties, surveys and measurements, damage typology, damage triggering events and possible interventions. The system supports searching features typical of standard databases, as it allows for the digitalization of a wide range of information including professional reports, books, articles and scientific papers. It also allows for computer aided retrieval of information tailored to user's requirements. The foreseen outputs will include a web user interface and a mobile application for visual inspection purposes.

  14. Algal culture studies related to a closed ecological life support system

    NASA Technical Reports Server (NTRS)

    Radmer, R.; Behrens, P.; Fernandez, E.; Ollinger, O.; Howell, C.

    1984-01-01

    Studies on the steady-state long-term (4 month) culture of Scenedesmus obliquus algae, maintained in an annular air-lift column operated as a turbidostat, were carried out to evaluate the life-supporting possibilities of this system. Chlorophyll production and cell number as functions of the dry weight were linear at constant illumination. Productivity (measured as the product of dry weight, mg/ml, and the growth rate, ml/hr) vs. dry weight rose linearly until the cell density reached a level at which light became limiting (89 percent absorption of the photosynthetically active radiation). In the initial, linear portion of the curve, the productivity was limited by cell growth at the given light intensity. The maximum dilution rate of the system corresponded to the doubling time of 13.4 hr, about half the maximum rate, with a productivity of 80 percent of the maximum theoretical productivity. The high light utilization efficiencies were contributed by the low (10 percent of full sunlight) incident intensities.

  15. 3D cell culture systems modeling tumor growth determinants in cancer target discovery.

    PubMed

    Thoma, Claudio R; Zimmermann, Miriam; Agarkova, Irina; Kelm, Jens M; Krek, Wilhelm

    2014-04-01

    Phenotypic heterogeneity of cancer cells, cell biological context, heterotypic crosstalk and the microenvironment are key determinants of the multistep process of tumor development. They sign responsible, to a significant extent, for the limited response and resistance of cancer cells to molecular-targeted therapies. Better functional knowledge of the complex intra- and intercellular signaling circuits underlying communication between the different cell types populating a tumor tissue and of the systemic and local factors that shape the tumor microenvironment is therefore imperative. Sophisticated 3D multicellular tumor spheroid (MCTS) systems provide an emerging tool to model the phenotypic and cellular heterogeneity as well as microenvironmental aspects of in vivo tumor growth. In this review we discuss the cellular, chemical and physical factors contributing to zonation and cellular crosstalk within tumor masses. On this basis, we further describe 3D cell culture technologies for growth of MCTS as advanced tools for exploring molecular tumor growth determinants and facilitating drug discovery efforts. We conclude with a synopsis on technological aspects for on-line analysis and post-processing of 3D MCTS models. PMID:24636868

  16. A biofidelic 3D culture model to study the development of brain cellular systems

    PubMed Central

    Ren, M.; Du, C.; Herrero Acero, E.; Tang-Schomer, M. D.; Özkucur, N.

    2016-01-01

    Little is known about how cells assemble as systems during corticogenesis to generate collective functions. We built a neurobiology platform that consists of fetal rat cerebral cortical cells grown within 3D silk scaffolds (SF). Ivermectin (Ivm), a glycine receptor (GLR) agonist, was used to modulate cell resting membrane potential (Vmem) according to methods described in a previous work that implicated Ivm in the arrangement and connectivity of cortical cell assemblies. The cells developed into distinct populations of neuroglial stem/progenitor cells, mature neurons or epithelial-mesenchymal cells. Importantly, the synchronized electrical activity in the newly developed cortical assemblies could be recorded as local field potential (LFP) measurements. This study therefore describes the first example of the development of a biologically relevant cortical plate assembly outside of the body. This model provides i) a preclinical basis for engineering cerebral cortex tissue autografts and ii) a biofidelic 3D culture model for investigating biologically relevant processes during the functional development of cerebral cortical cellular systems. PMID:27112667

  17. Fluorescence-based system for measurement of electrophysiological changes in stretched cultured cardiomyocytes.

    PubMed

    Duverger, James E; Béland, Jonathan; Maguy, Ange; Adegbindin, Mouhamed M; Comtois, Philippe

    2011-01-01

    Acute or sustained stretch of cardiac tissue is known to play a key role in arrhythmogenesis. Using a fluorescence approach, we designed a system measuring calcium transients and transmembrane potential changes in monolayers of cultured cardiomyocytes under uniaxial elongation and electrical stimulation. Cardiac myocytes are seeded on a rectangular PDMS template held and stretched by a motorized linear guide system. Electrical stimulation is performed with two parallel carbon electrodes supplied by amplified pulses from a digital-to-analog converter. The cells are stained with either voltage- or calcium-sensitive dye (di-4-ANEPPS and Fluo-4 AM respectively). The two available excitation light sources are both current-controlled LED arrays (λ = 523 ± 45 nm for di-4-ANEPPS and λ = 505 ± 15 nm for Fluo-4 AM). The filtered emitted fluorescence (λ > 610 nm for di-4-ANEPPS and λ = 535 ± 25 nm for Fluo-4 AM) is transduced to current with a photodiode, converted to amplified voltage signals and digitized. The design and preliminary validation results are presented. PMID:22254244

  18. Cultural Heritage: An example of graphical documentation with automated photogrammetric systems

    NASA Astrophysics Data System (ADS)

    Giuliano, M. G.

    2014-06-01

    In the field of Cultural Heritage, the use of automated photogrammetric systems, based on Structure from Motion techniques (SfM), is widely used, in particular for the study and for the documentation of the ancient ruins. This work has been carried out during the PhD cycle that was produced the "Carta Archeologica del territorio intorno al monte Massico". The study suggests the archeological documentation of the mausoleum "Torre del Ballerino" placed in the south-west area of Falciano del Massico, along the Via Appia. The graphic documentation has been achieved by using photogrammetric system (Image Based Modeling) and by the classical survey with total station, Nikon Nivo C. The data acquisition was carried out through digital camera Canon EOS 5D Mark II with Canon EF 17-40 mm f/4L USM @ 20 mm with images snapped in RAW and corrected in Adobe Lightroom. During the data processing, the camera calibration and orientation was carried out by the software Agisoft Photoscans and the final result has allowed to achieve a scaled 3D model of the monument, imported in software MeshLab for the different view. Three orthophotos in jpg format were extracted by the model, and then were imported in AutoCAD obtaining façade's surveys.

  19. Evaluation of Differentiated Human Bronchial Epithelial Cell Culture Systems for Asthma Research

    PubMed Central

    Stewart, Ceri E.; Torr, Elizabeth E.; Mohd Jamili, Nur H.; Bosquillon, Cynthia; Sayers, Ian

    2012-01-01

    The aim of the current study was to evaluate primary (human bronchial epithelial cells, HBEC) and non-primary (Calu-3, BEAS-2B, BEAS-2B R1) bronchial epithelial cell culture systems as air-liquid interface- (ALI-) differentiated models for asthma research. Ability to differentiate into goblet (MUC5AC+) and ciliated (β-Tubulin IV+) cells was evaluated by confocal imaging and qPCR. Expression of tight junction/adhesion proteins (ZO-1, E-Cadherin) and development of transepithelial electrical resistance (TEER) were assessed. Primary cells showed localised MUC5AC, β-Tubulin IV, ZO-1, and E-Cadherin and developed TEER with, however, a large degree of inter- and intradonor variation. Calu-3 cells developed a more reproducible TEER and a phenotype similar to primary cells although with diffuse β-Tubulin IV staining. BEAS-2B cells did not differentiate or develop tight junctions. These data highlight the challenges in working with primary cell models and the need for careful characterisation and selection of systems to answer specific research questions. PMID:22287976

  20. Twenty-four well plate miniature bioreactor system as a scale-down model for cell culture process development.

    PubMed

    Chen, Aaron; Chitta, Rajesh; Chang, David; Amanullah, Ashraf

    2009-01-01

    Increasing the throughput and efficiency of cell culture process development has become increasingly important to rapidly screen and optimize cell culture media and process parameters. This study describes the application of a miniaturized bioreactor system as a scaled-down model for cell culture process development using a CHO cell line expressing a recombinant protein. The microbioreactor system (M24) provides non-invasive online monitoring and control capability for process parameters such as pH, dissolved oxygen (DO), and temperature at the individual well level. A systematic evaluation of the M24 for cell culture process applications was successfully completed. Several challenges were initially identified. These included uneven gas distribution in the wells due to system design and lot to lot variability, foaming issues caused by sparging required for active DO control, and pH control limitation under conditions of minimal dissolved CO2. A high degree of variability was found which was addressed by changes in the system design. The foaming issue was resolved by addition of anti-foam, reduction of sparge rate, and elimination of DO control. The pH control limitation was overcome by a single manual liquid base addition. Intra-well reproducibility, as indicated by measurements of process parameters, cell growth, metabolite profiles, protein titer, protein quality, and scale-equivalency between the M24 and 2 L bioreactor cultures were very good. This evaluation has shown feasibility of utilizing the M24 as a scale-down tool for cell culture application development under industrially relevant process conditions. PMID:18683260

  1. Modeling inflammation and oxidative stress in gastrointestinal disease development using novel organotypic culture systems.

    PubMed

    Hartman, Kira G; Bortner, James D; Falk, Gary W; Yu, Jian; Martín, Martín G; Rustgi, Anil K; Lynch, John P

    2013-01-01

    Gastroesophageal reflux disease (GERD), Barrett's esophagus (BE), graft-versus-host disease (GVHD), and inflammatory bowel diseases such as ulcerative colitis and Crohn's disease are common human gastrointestinal diseases that share inflammation as a key driver for their development. A general outcome resulting from these chronic inflammatory conditions is increased oxidative stress. Oxidative stress is caused by the generation of reactive oxygen and nitrogen species that are part of the normal inflammatory response, but are also capable of damaging cellular DNA, protein, and organelles. Damage to DNA can include DNA strand breaks, point mutations due to DNA adducts, as well as alterations in methylation patterns leading to activation of oncogenes or inactivation of tumor suppressors. There are a number of significant long-term consequences associated with chronic oxidative stress, most notably cancer. Infiltrating immune cells and stromal components of tissue including fibroblasts contribute to dynamic changes occurring in tissue related to disease development. Immune cells can potentiate oxidative stress, and fibroblasts have the capacity to contribute to advanced growth and proliferation of the epithelium and any resultant cancers. Disease models for GERD, BE, GVHD, and ulcerative colitis based on three-dimensional human cell and tissue culture systems that recapitulate in vivo growth and differentiation in inflammatory-associated microphysiological environments would enhance our understanding of disease progression and improve our ability to test for disease-prevention strategies. The development of physiologically relevant, human cell-based culture systems is therefore a major focus of our research. These novel models will be of enormous value, allowing us to test hypotheses and advance our understanding of these disorders, and will have a translational impact allowing us to more rapidly develop therapeutic and chemopreventive agents. In summary, this work to

  2. Effects of Crude Oil, Dispersant, and Oil-Dispersant Mixtures on Human Fecal Microbiota in an In Vitro Culture System

    PubMed Central

    Kim, Jong Nam; Kim, Bong-Soo; Kim, Seong-Jae; Cerniglia, Carl E.

    2012-01-01

    ABSTRACT The Deepwater Horizon oil spill of 2010 raised concerns that dispersant and dispersed oil, as well as crude oil itself, could contaminate shellfish and seafood habitats with hazardous residues that had potential implications for human health and the ecosystem. However, little is known about the effects of crude oil and dispersant on the human fecal microbiota. The aim of this research was to evaluate the potential effects of Deepwater Horizon crude oil, Corexit 9500 dispersant, and their combination on human fecal microbial communities, using an in vitro culture test system. Fecal specimens from healthy adult volunteers were made into suspensions, which were then treated with oil, dispersant, or oil-dispersant mixtures under anaerobic conditions in an in vitro culture test system. Perturbations of the microbial community, compared to untreated control cultures, were assessed using denaturing gradient gel electrophoresis (DGGE), real-time PCR, and pyrosequencing methods. DGGE and pyrosequencing analysis showed that oil-dispersant mixtures reduced the diversity of fecal microbiota from all individuals. Real-time PCR results indicated that the copy numbers of 16S rRNA genes in cultures treated with dispersed oil or oil alone were significantly lower than those in control incubations. The abundance of the Bacteroidetes decreased in crude oil-treated and dispersed-oil-treated cultures, while the Proteobacteria increased in cultures treated with dispersed oil. In conclusion, the human fecal microbiota was affected differently by oil and dispersed oil, and the influence of dispersed oil was significantly greater than that of either oil or dispersant alone compared to control cultures. PMID:23093387

  3. In vitro culture system to determine MICs and MBCs of antimicrobial agents against Treponema pallidum subsp. pallidum (Nichols strain).

    PubMed Central

    Norris, S J; Edmondson, D G

    1988-01-01

    A new procedure for determining the susceptibility of Treponema pallidum subsp. pallidum to antimicrobial agents was developed, utilizing a tissue culture system which promotes the in vitro multiplication of this organism. In the absence of antibiotics, T. pallidum (Nichols virulent strain) multiplied an average of 10-fold when incubated for 7 days in the presence of Sf1Ep cottontail rabbit epithelial cell cultures. Varied concentrations of penicillin G, tetracycline, erythromycin, and spectinomycin were added to triplicate cultures to determine their effects on treponemal multiplication, motility, and virulence. The MIC of each antibiotic was defined as the lowest concentration which prevented treponemal multiplication, whereas the MBC was defined as the lowest concentration which abrogated the ability of the cultured treponemes to multiply and cause lesions in rabbits. The in vitro culture technique provided highly reproducible MICs and (in parentheses) MBCs of each of the antibiotics tested: aqueous penicillin G, 0.0005 (0.0025) microgram/ml; tetracycline, 0.2 (0.5) microgram/ml; erythromycin, 0.005 (0.005) microgram/ml; and spectinomycin, 0.5 (0.5) microgram/ml. The significance of these results in light of the in vivo activities and the previous in vitro evaluations of these antibiotics is discussed. The T. pallidum in vitro cultivation system shows promise as a method for studying the interaction between T. pallidum and antimicrobial agents and for screening new antibiotics for syphilis therapy. PMID:2964810

  4. An Efficacious Measurement of Learning Initiatives: E-Learning Systems, Learning-Organization Culture, Knowledge Creation, and Innovativeness

    ERIC Educational Resources Information Center

    Grundhoefer, Raymie

    2013-01-01

    The purpose of this research is twofold: (a) develop a validated measure for learning initiatives based on knowledge-creation theory and (b) conduct a quantitative study to investigate the relationships between electronic learning systems, learning-organization culture, efficacious knowledge creation (EKC), and innovativeness. Although Cheng-Chang…

  5. Design of a Basic System of Indicators for Monitoring and Evaluating Spanish Cooperation's Culture and Development Strategy

    ERIC Educational Resources Information Center

    Coll-Serrano, Vicente; Carrasco-Arroyo, Salvador; Blasco-Blasco, Olga; Vila-Lladosa, Luis

    2012-01-01

    Objective: This article describes the process implemented in order to define and build up a Basic Monitoring and Evaluation System for Spanish Cooperation's Culture and Development Strategy (CD-S). Research Design: Delphi techniques were used to assess a wide catalogue of indicators for each of the strategic areas included in the CD-S. Afterward,…

  6. A NOVEL SYSTEM FOR THE CO-CULTURE OF EPIDIDYMAL EPITHELIAL CELLS AND SPERM FROM ADULT RATS

    EPA Science Inventory

    To study interactions which occur between the epididymal epithelial cells and sperm within the epididymis during sperm maturation, a specialized co-culture system capable of supporting the differentiated function of these cell types must be utilized. A multifaceted approach has b...

  7. Forensic, Cultural, and Systems Issues in Child Sexual Abuse Cases--Part 2: Research and Practitioner Issues

    ERIC Educational Resources Information Center

    Tishelman, Amy C.; Geffner, Robert

    2010-01-01

    This article introduces the second issue of the special double issue focusing on forensic, cultural, and systems issues in child sexual abuse cases. We briefly review the articles, which include a discussion of child sexual abuse myths, an empirical analysis of extended child sexual abuse evaluations, an article on the role of the medical provider…

  8. The Effects of the ARC Organizational Intervention on Caseworker Turnover, Climate, and Culture in Children's Service Systems

    ERIC Educational Resources Information Center

    Glisson, Charles; Dukes, Denzel; Green, Philip

    2006-01-01

    Objective: This study examines the effects of the Availability, Responsiveness, and Continuity (ARC) organizational intervention strategy on caseworker turnover, climate, and culture in a child welfare and juvenile justice system. Method: Using a pre-post, randomized blocks, true experimental design, 10 urban and 16 rural case management teams…

  9. Systemic Transformation from the Ground-Up: Using Learning Lab to Design Culturally Responsive Schoolwide Positive Behavioral Supports

    ERIC Educational Resources Information Center

    Bal, Aydin; Kozleski, Elizabeth B.; Schrader, Elizabeth M.; Rodriguez, Esmeralda M.; Pelton, Scott

    2014-01-01

    The enduring existence of disproportionate representation of students from culturally and linguistically diverse backgrounds in special education programs and disciplinary practices creates a double bind for educators, educational leaders, and families. Disproportionality is an adaptive systemic issue that is not under any entity's control;…

  10. Growth performance, fillet quality, and reproductive maturity of rainbow trout (Oncorhynchus mykiss) cultured to 5 kilograms within freshwater recirculating systems

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rainbow trout are commonly cultured within aquaculture systems to one pound or less and marketed as pan-sized fillets. Production of larger rainbow trout provides a distinguishable product. Research that describes the growth performance and fillet quality of large rainbow trout is limited, particula...

  11. An Introduction to the System and Culture of the College Entrance Examination of China. Research Notes. RN-28

    ERIC Educational Resources Information Center

    Wang, Xiang Bo

    2006-01-01

    This report provides an introduction to the system and culture of the college entrance examination (CEE) of China. College entrance examinations in China (and in several other Asian countries such as Korea and Japan) are so important that they are commonly dubbed "once in a lifetime" or a "one-test-to-determine-a-life" for most…

  12. Hyperhydricity and flavonoid content of Scutellaria species in vitro on polyester-supported liquid culture systems

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Three Scutellaria species (Scutellaria lateriflora, S. costaricana and S. baicalensis) were grown in different in vitro physical environments: agar, liquid culture, and liquid culture with fiber-supported paper (with initial media volumes of 20 mL and 30 mL). During an eight-week time course, tiss...

  13. Cross Cultural Analysis of the Use and Perceptions of Web-Based Learning Systems

    ERIC Educational Resources Information Center

    Arenas-Gaitan, Jorge; Ramirez-Correa, Patricio E.; Rondan-Cataluna, F. Javier

    2011-01-01

    The main objective of this paper is to examine cultural differences and technology acceptances from students of two universities, one is from a European country: Spain, and the other is in Latin America: Chile. Both of them provide their students with e-learning platforms. The technology acceptance model (TAM) and Hofstede's cultural dimensions…

  14. Universality and Cultural Diversity in Professional Ethical Development: From Kohlberg to Dynamic Systems Theory

    ERIC Educational Resources Information Center

    Kim, Minkang

    2012-01-01

    Upholding ethical standards is part of what it means to be a professional and therefore part of professional education, but to what extent is the development of ethical reasoning universal across cultures, or is it highly dependent on culture? If universal, how can we explain the unique patterns of moral reasoning and behaviour in Asia, which…

  15. Apical polarity in three-dimensional culture systems: where to now?

    SciTech Connect

    Inman, J.L.; Bissell, Mina

    2010-01-21

    Delineation of the mechanisms that establish and maintain the polarity of epithelial tissues is essential to understanding morphogenesis, tissue specificity and cancer. Three-dimensional culture assays provide a useful platform for dissecting these processes but, as discussed in a recent study in BMC Biology on the culture of mammary gland epithelial cells, multiple parameters that influence the model must be taken into account.

  16. Treatment of swine wastewater using a saturated-soil-culture soybean and flooded rice system

    SciTech Connect

    Szoegi, A.A.; Hunt, P.G.; Humenik, F.J.

    2000-04-01

    Constructed wetlands have potential for treatment of livestock wastewater, but they generally contain wetland plants rather than agronomic crops. The authors evaluated two agronomic crops, saturated-soil-culture (SSC) soybean and flooded rice, in a constructed wetland system used for swine wastewater treatment. Both crop production and treatment efficiency were evaluated from 1993 to 1996 in two 4-m x 33.5-m constructed wetland cells that were connected in series. The first cell contained SSC soybean--four cultivars planted in a randomized complete block design with four replications. Flooded rice Maybelle was planted in the second cell. From the first to fourth year, wastewater application rates were gradually increased to obtain rates of 2.0 to 8.8 and 0.5 to 2.2 kg/ha d for total N and P, respectively. The best soybean grain and dry matter yields were 4.0 and 9.1 Mg/ha, respectively. These were obtained with soybean Young at the lowest wastewater application rate. Increasing total N loading rates and the associated higher NH{sub 4}-N concentrations depressed soybean seed yield and dry matter production. On the other hand, both rice grain and dry matter production were stable over the application range; mean values were 4.0 and 10.9 Mg ha{sup {minus}1}, respectively. Nutrient mass reductions were good; removal values increased linearly with loading rates (y = 0.69N load + 0.45, R{sup 2} = 0.99 and y = 0.45P load + 0.20, R{sup 2} = 0.95). At the highest loading rate, the system removed 751 and 156 kg/ha yr N and P, respectively. It appears that the SSC soybean and flooded rice system could be useful for liquid manure management in confined livestock production. The system produced comparable treatment to systems with natural wetland plants; moreover, the soybean and rice are marketable crops. However, the flooded rice seems to be the more robust component for high wastewater application rates.

  17. Enhanced Propionate Formation by Propionibacterium freudenreichii subsp. freudenreichii in a Three-Electrode Amperometric Culture System

    PubMed Central

    Emde, Rainer; Schink, Bernhard

    1990-01-01

    In order to influence the fermentation pattern of Propionibacterium freudenreichii towards enhanced propionate formation, growth and product formation with glucose and lactate as energy sources were studied in a three-electrode poised-potential amperometric culture system. With anthraquinone 2,6-disulfonic acid (E0′ = −184 mV; poised electron potential = −224 mV) or cobalt sepulchrate (E0′ = −350 mV; −390 mV) as mediator and an activated platinum working electrode, reduction of bacterially oxidized mediator occurred fast enough to keep more than 50% of the respective mediator (in minimum 0.4 mM) in the reduced state, up to a current of 2 mA. With glucose as substrate, 90.0 or 97.3% propionate was formed during exponential growth in the presence of 0.5 mM anthraquinone 2,6-disulfonic acid or 0.4 mM cobalt sepulchrate, respectively. Growth yields of 56.3 or 53.8 g of cell material per mol of substrate degraded were calculated, respectively, and the electrons were transferred quantitatively from the working electrode to the bacterial cells. With l-lactate, only 68.6 or 72.9% propionate was formed with the same mediators. The results are discussed with respect to energetics, electron transfer potentials, and potential application of the new technique in technical propionate production. PMID:16348285

  18. Enhanced Propionate Formation by Propionibacterium freudenreichii subsp. freudenreichii in a Three-Electrode Amperometric Culture System.

    PubMed

    Emde, R; Schink, B

    1990-09-01

    In order to influence the fermentation pattern of Propionibacterium freudenreichii towards enhanced propionate formation, growth and product formation with glucose and lactate as energy sources were studied in a three-electrode poised-potential amperometric culture system. With anthraquinone 2,6-disulfonic acid (E(0)' = -184 mV; poised electron potential = -224 mV) or cobalt sepulchrate (E(0)' = -350 mV; -390 mV) as mediator and an activated platinum working electrode, reduction of bacterially oxidized mediator occurred fast enough to keep more than 50% of the respective mediator (in minimum 0.4 mM) in the reduced state, up to a current of 2 mA. With glucose as substrate, 90.0 or 97.3% propionate was formed during exponential growth in the presence of 0.5 mM anthraquinone 2,6-disulfonic acid or 0.4 mM cobalt sepulchrate, respectively. Growth yields of 56.3 or 53.8 g of cell material per mol of substrate degraded were calculated, respectively, and the electrons were transferred quantitatively from the working electrode to the bacterial cells. With l-lactate, only 68.6 or 72.9% propionate was formed with the same mediators. The results are discussed with respect to energetics, electron transfer potentials, and potential application of the new technique in technical propionate production. PMID:16348285

  19. Studies on the porphobilinogen deaminase–uroporphyrinogen cosynthetase system of cultured soya-bean cells

    PubMed Central

    Llambías, Elena B. C.; Batlle, Alcira M. Del C.

    1971-01-01

    1. Porphobilinogenase was isolated and purified from soya-bean callus tissue; its components, porphobilinogen deaminase and uroporphyrinogen isomerase, were separated and purified. 2. The purified porphobilinogenase was resolved into two bands on starch-gel electrophoresis. The molecular weights of porphobilinogenase, deaminase and isomerase fractions were determined by the gel-filtration method. Porphobilinogenase activity was affected by the presence of air; uroporphyrinogens were only formed under anaerobic conditions, although substrate consumption was the same in the absence of oxygen as in its presence. 3. pH-dependence of both porphobilinogenase and deaminase was the same and a sharp optimum at pH 7.2 was obtained. Isomerase was heat-labile, but the presence of ammonium ions or porphobilinogen afforded some protection against inactivation. The action of several compounds added to the system was studied. Cysteine, thioglycollate, ammonium ions and hydroxylamine inhibited porphobilinogenase; certain concentrations of sodium and magnesium salts enhanced activity; some dicarboxylic acids and 2-methoxy-5-nitrotropone inhibited the deaminase. 4. δ-Aminolaevulate and ethionine in the culture media stimulated porphyrin synthesis and increased porphobilinogenase activity, whereas iron deficiency resulted in porphyrin accumulation. 5. The development of chlorophyll and porphobilinogenase on illumination of dark-grown callus was followed. 6. A hypothetical scheme is suggested for the enzymic synthesis of uroporphyrinogens from porphobilinogen. PMID:5165654

  20. Oxygen Limited Bioreactors System For Nitrogen Removal Using Immobilized Mix Culture

    NASA Astrophysics Data System (ADS)

    Pathak, B. K.; Sumino, T.; Saiki, Y.; Kazama, F.

    2005-12-01

    Recently nutrients concentrations especially nitrogen in natural water is alarming in the world wide. Most of the effort is being done on the removal of high concentration of nitrogen especially from the wastewater treatment plants. The removal efficiency is targeted in all considering the effluent discharge standard set by the national environment agency. In many cases, it does not meet the required standard and receiving water is being polluted. Eutrophication in natural water bodies has been reported even if the nitrogen concentration is low and self purification of natural systems itself is not sufficient to remove the nitrogen due to complex phenomenon. In order to recover the pristine water environment, it is very essential to explore bioreactor systems for natural water systems using immobilized mix culture. Microorganism were entrapped in Polyethylene glycol (PEG) prepolymer gel and cut into 3mm cubic immobilized pellets. Four laboratory scale micro bio-reactors having 0.1 L volumes were packed with immobilized pellets with 50% compact ratio. RUN1, RUN2, RUN3 and RUN4 were packed with immobilized pellets from reservoirs sediments, activated sludge (AS), mixed of AS, AG and biodegradable plastic and anaerobic granules (AG) respectively. Water from Shiokawa Reservoirs was feed to all reactors with supplemental ammonia and nitrite nitrogen as specified in the results and discussions. The reactors were operated dark incubated room in continuous flow mode with hydraulic retention time of 12 hours under oxygen limiting condition. Ammonium, nitrate nitrite nitrogen and total organic carbon (TOC) concentrations were measured as described in APWA and AWWA (1998). Laboratory scale four bioreactors containing different combination of immobilized cell were monitored for 218 days. Influent NH4+-N and NO2--N concentration were 2.27±0.43 and 2.05±0.41 mg/l respectively. Average dissolved oxygen concentration and pH in the reactors were 0.40-2.5 mg/l and pH 6

  1. Technology Transfer Challenges: A Case Study of User-Centered Design in NASA's Systems Engineering Culture

    NASA Technical Reports Server (NTRS)

    Quick, Jason

    2009-01-01

    The Upper Stage (US) section of the National Aeronautics and Space Administration's (NASA) Ares I rocket will require internal access platforms for maintenance tasks performed by humans inside the vehicle. Tasks will occur during expensive critical path operations at Kennedy Space Center (KSC) including vehicle stacking and launch preparation activities. Platforms must be translated through a small human access hatch, installed in an enclosed worksite environment, support the weight of ground operators and be removed before flight - and their design must minimize additional vehicle mass at attachment points. This paper describes the application of a user-centered conceptual design process and the unique challenges encountered within NASA's systems engineering culture focused on requirements and "heritage hardware". The NASA design team at Marshall Space Flight Center (MSFC) initiated the user-centered design process by studying heritage internal access kits and proposing new design concepts during brainstorming sessions. Simultaneously, they partnered with the Technology Transfer/Innovative Partnerships Program to research inflatable structures and dynamic scaffolding solutions that could enable ground operator access. While this creative, technology-oriented exploration was encouraged by upper management, some design stakeholders consistently opposed ideas utilizing novel, untested equipment. Subsequent collaboration with an engineering consulting firm improved the technical credibility of several options, however, there was continued resistance from team members focused on meeting system requirements with pre-certified hardware. After a six-month idea-generating phase, an intensive six-week effort produced viable design concepts that justified additional vehicle mass while optimizing the human factors of platform installation and use. Although these selected final concepts closely resemble heritage internal access platforms, challenges from the application of the

  2. Multimedia systems for art and culture: a case study of Brihadisvara Temple

    NASA Astrophysics Data System (ADS)

    Jain, Anil K.; Goel, Sanjay; Agarwal, Sachin; Mittal, Vipin; Sharma, Hariom; Mahindru, Ranjeev

    1997-01-01

    In India a temple is not only a structure of religious significance and celebration, but it also plays an important role in the social, administrative and cultural life of the locality. Temples have served as centers for learning Indian scriptures. Music and dance were fostered and performed in the precincts of the temples. Built at the end of the 10th century, the Brihadisvara temple signified new design methodologies. We have access to a large number of images, audio and video recordings, architectural drawings and scholarly publications of this temple. A multimedia system for this temple is being designed which is intended to be used for the following purposes: (1) to inform and enrich the general public, and (2) to assist the scholars in their research. Such a system will also preserve and archive old historical documents and images. The large database consists primarily of images which can be retrieved using keywords, but the emphasis here is largely on techniques which will allow access using image content. Besides classifying images as either long shots or close-ups, deformable template matching is used for shape-based query by image content, and digital video retrieval. Further, to exploit the non-linear accessibility of video sequences, key frames are determined to aid the domain experts in getting a quick preview of the video. Our database also has images of several old, and rare manuscripts many of which are noisy and difficult to read. We have enhanced them to make them more legible. We are also investigating the optimal trade-off between image quality and compression ratios.

  3. Communicating the Science of the Earth System Through Arts and Culture to Reach Broad Audiences

    NASA Astrophysics Data System (ADS)

    Gardiner, L.; Genyuk, J.; Bergman, J.; Johnson, R.; Foster, S.; Hatheway, B.; Russell, R.

    2008-12-01

    Links between the science of Earth and the visual and literary arts, cultures, and human history provides important context and connections for learners of all ages. Several new features that foster a multidisciplinary approach to learning about our planet are now available on Windows to the Universe (www.windows.ucar.edu), an educational Web site that includes over 6000 pages of content and is used by over 20 million people each year. The Clouds in Art interactive encourages users to identify cloud types depicted in well-known landscape paintings. Examples of poems by historic poets describe weather phenomena and link to information about the science of weather. A new feature allows users to post their original poetry about an image of weather phenomena. Historic image collections emphasize human connections to the Earth system. For example, a collection of images that visually describes Inuit traditions is linked to Web content about Earth's polar regions and the impact of climate change in the Arctic. To support K-12 classroom learning of Earth system concepts and engage visual learners, several new classroom activities make use of photographs, satellite images, and animations of remote sensing data. In one activity, students learn about the impact of climate change in the Arctic by working with photographs of Alaskan glaciers taken over the past century. These new interdisciplinary features on Windows to the Universe, combined with a wealth of existing content on the site about the history of science and mythology, provide other ways to appreciate science phenomena as well as alternate avenues into science for the general public, teachers and students. Windows to the Universe, a project of the University Corporation for Atmospheric Research Office of Education and Outreach, provides users with content about the Earth and space sciences at three levels of instruction in both English and Spanish.

  4. Hematopoietic Stem Cells, Their Niche, and the Concept of Co-Culture Systems: A Critical Review.

    PubMed

    Vaidya, Anuradha; Kale, Vaijayanti

    2015-01-01

    Hematopoietic stem cells (HSCs) have the ability to self-renew and give rise to all lineages of blood cells. HSCs reside in niches that are local tissue microenvironments that maintain and regulate them. Although much progress has been made in elucidating the location and cellular components of the HSC niche, however it still remains incompletely defined. Transplantation using HSCs has been applied for the treatment of several diseases but with limited success. Furthermore, although human HSC transplantation has been widely used to rescue the patients after cytoablative therapies, quantitative in vivo human assays for hematopoietic cells have been considered to be neither ethical nor practical. Since HSCs persist in small quantities in the body, understanding the mechanism that govern their fate is essential for the advancement of HSC expansion and transplantation in the future. Since bone marrow is the primary site of HSC maintenance and hematopoiesis, defining the niche components that work in concert to regulate hematopoiesis is crucial to improve regeneration following injury or following HSC transplantation and to also understand how disordered niche function could contribute to disease. In recent years, there has been a growing realization of the limitations in identifying the primitive HSCs by its phenotype alone and therefore the concept of co-culture systems (functional in vitro assays) has become increasingly important to demonstrate the presence of primitive hematopoietic cells by estimating their biological functions. This system has provided a basis for the development of powerful assay procedures for expanding, quantitating and distinguishing cells at discrete stages of early hematopoietic cell differentiation. PMID:26665935

  5. A Cell-Free Translocation System Using Extracts of Cultured Insect Cells to Yield Functional Membrane Proteins

    PubMed Central

    Ezure, Toru; Nanatani, Kei; Sato, Yoko; Suzuki, Satomi; Aizawa, Keishi; Souma, Satoshi; Ito, Masaaki; Hohsaka, Takahiro; von Heijine, Gunnar; Utsumi, Toshihiko; Abe, Keietsu; Ando, Eiji; Uozumi, Nobuyuki

    2014-01-01

    Cell-free protein synthesis is a powerful method to explore the structure and function of membrane proteins and to analyze the targeting and translocation of proteins across the ER membrane. Developing a cell-free system based on cultured cells for the synthesis of membrane proteins could provide a highly reproducible alternative to the use of tissues from living animals. We isolated Sf21 microsomes from cultured insect cells by a simplified isolation procedure and evaluated the performance of the translocation system in combination with a cell-free translation system originating from the same source. The isolated microsomes contained the basic translocation machinery for polytopic membrane proteins including SRP-dependent targeting components, translocation channel (translocon)-dependent translocation, and the apparatus for signal peptide cleavage and N-linked glycosylation. A transporter protein synthesized with the cell-free system could be functionally reconstituted into a lipid bilayer. In addition, single and double labeling with non-natural amino acids could be achieved at both the lumen side and the cytosolic side in this system. Moreover, tail-anchored proteins, which are post-translationally integrated by the guided entry of tail-anchored proteins (GET) machinery, were inserted correctly into the microsomes. These results showed that the newly developed cell-free translocation system derived from cultured insect cells is a practical tool for the biogenesis of properly folded polytopic membrane proteins as well as tail-anchored proteins. PMID:25486605

  6. The alc-GR system: a modified alc gene switch designed for use in plant tissue culture.

    PubMed

    Roberts, Gethin R; Garoosi, G Ali; Koroleva, Olga; Ito, Masaki; Laufs, Patrick; Leader, David J; Caddick, Mark X; Doonan, John H; Tomsett, A Brian

    2005-07-01

    The ALCR/alcA (alc) two-component, ethanol-inducible gene expression system provides stringent control of transgene expression in genetically modified plants. ALCR is an ethanol-activated transcription factor that can drive expression from the ALCR-responsive promoter (alcA). However, the alc system has been shown to have constitutive expression when used in plant callus or cell suspension cultures, possibly resulting from endogenous inducer produced in response to lowered oxygen availability. To widen the use of the alc system in plant cell culture conditions, the receptor domain of the rat glucocorticoid receptor (GR) was translationally fused to the C terminus of ALCR to produce ALCR-GR, which forms the basis of a glucocorticoid-inducible system (alc-GR). The alc-GR switch system was tested in tobacco (Nicotiana tabacum) Bright Yellow-2 suspension cells using a constitutively expressed ALCR-GR with four alternative alcA promoter-driven reporter genes: beta-glucuronidase, endoplasmic reticulum-targeted green fluorescent protein, haemagglutinin, and green fluorescent protein-tagged Arabidopsis (Arabidopsis thaliana) Arath;CDKA;1 cyclin-dependent kinase. Gene expression was shown to be stringently dependent on the synthetic glucocorticoid dexamethasone and, in cell suspensions, no longer required ethanol for induction. Thus, the alc-GR system allows tight control of alcA-driven genes in cell culture and complements the conventional ethanol switch used in whole plants. PMID:16010000

  7. The alc-GR System. A Modified alc Gene Switch Designed for Use in Plant Tissue Culture1[w

    PubMed Central

    Roberts, Gethin R.; Garoosi, G. Ali; Koroleva, Olga; Ito, Masaki; Laufs, Patrick; Leader, David J.; Caddick, Mark X.; Doonan, John H.; Tomsett, A. Brian

    2005-01-01

    The ALCR/alcA (alc) two-component, ethanol-inducible gene expression system provides stringent control of transgene expression in genetically modified plants. ALCR is an ethanol-activated transcription factor that can drive expression from the ALCR-responsive promoter (alcA). However, the alc system has been shown to have constitutive expression when used in plant callus or cell suspension cultures, possibly resulting from endogenous inducer produced in response to lowered oxygen availability. To widen the use of the alc system in plant cell culture conditions, the receptor domain of the rat glucocorticoid receptor (GR) was translationally fused to the C terminus of ALCR to produce ALCR-GR, which forms the basis of a glucocorticoid-inducible system (alc-GR). The alc-GR switch system was tested in tobacco (Nicotiana tabacum) Bright Yellow-2 suspension cells using a constitutively expressed ALCR-GR with four alternative alcA promoter-driven reporter genes: β-glucuronidase, endoplasmic reticulum-targeted green fluorescent protein, haemagglutinin, and green fluorescent protein-tagged Arabidopsis (Arabidopsis thaliana) Arath;CDKA;1 cyclin-dependent kinase. Gene expression was shown to be stringently dependent on the synthetic glucocorticoid dexamethasone and, in cell suspensions, no longer required ethanol for induction. Thus, the alc-GR system allows tight control of alcA-driven genes in cell culture and complements the conventional ethanol switch used in whole plants. PMID:16010000

  8. A New Experimental System for the Extended Application of Cyclic Hydrostatic Pressure to Cell Culture

    PubMed Central

    Maul, Timothy M.; Hamilton, Douglas W.; Nieponice, Alejandro; Soletti, Lorenzo

    2007-01-01

    Mechanical forces have been shown to be important stimuli for the determination and maintenance of cellular phenotype and function. Many cells are constantly exposed in vivo to cyclic pressure, shear stress, and/or strain. Therefore, the ability to study the effects of these stimuli in vitro is important for understanding how they contribute to both normal and pathologic states. While there exist commercial as well as custom-built devices for the extended application of cyclic strain and shear stress, very few cyclic pressure systems have been reported to apply stimulation longer than 48 h. However, pertinent responses of cells to mechanical stimulation may occur later than this. To address this limitation, we have designed a new cyclic hydrostatic pressure system based upon the following design variables: minimal size, stability of pressure and humidity, maximal accessibility, and versatility. Computational fluid dynamics (CFD) was utilized to predict the pressure and potential shear stress within the chamber during the first half of a 1.0 Hz duty cycle. To biologically validate our system, we tested the response of bone marrow progenitor cells (BMPCs) from Sprague Dawley rats to a cyclic pressure stimulation of 120/80 mm Hg, 1.0 Hz for 7 days. Cellular morphology was measured using Scion Image, and cellular proliferation was measured by counting nuclei in ten fields of view. CFD results showed a constant pressure across the length of the chamber and no shear stress developed at the base of the chamber where the cells are cultured. BMPCs from Sprague Dawley rats demonstrated a significant change in morphology versus controls by reducing their size and adopting a more rounded morphology. Furthermore, these cells increased their proliferation under cyclic hydrostatic pressure. We have demonstrated that our system imparts a single mechanical stimulus of cyclic hydrostatic pressure and is capable of at least 7 days of continuous operation without affecting cellular

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

    SciTech Connect

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

    1991-02-01

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

  10. Effect of Aflatoxin B1 on Growth of Bovine Mammary Epithelial Cells in 3D and Monolayer Culture System

    PubMed Central

    Forouharmehr, Ali; Harkinezhad, Taher; Qasemi-Panahi, Babak

    2013-01-01

    Purpose: Many studies have been showed transfer of aflatoxins, toxins produced by Aspergillus flvaus and Aspergillus parasiticus fungi, into milk. These toxins are transferred into the milk through digestive system by eating contaminated food. Due to the toxicity of these materials, it seems that it has side effects on the growth of mammary cells. Therefore, the present work aimed to investigate possible toxic effects of aflatoxin B1 (AFB1) on bovine mammary epithelial cells in monolayer and three-dimensional cultures. Methods: Specimens of the mammary tissue of bovine were sized out in size 2×2 cm in slaughterhouse. After disinfection and washing in sterile PBS, primary cell culture was performed by enzymatic digestion of tissue with collagenase. When proper numbers of cells were achieved in monolayer culture, cells were seeded in a 24-well culture plate for three-dimensional (3D) culture in Matrigel matrix. After 21 days of 3D culture and reaching the required number of cells, the concentrations of 15, 25 and 35 µL of AFB1 were added to the culture in quadruplicate and incubated for 8 hours. Cellular cytotoxicity was examined using standard colorimetric assay and finally, any change in the morphology of the cells was studied by microscopic technique. Results: Microscopic investigations showed necrosis of the AFB1-exposed cells compared to the control cells. Also, bovine mammary epithelial cells were significantly affected by AFB1 in dose and time dependent manner in cell viability assays. Conclusion: According to the results, it seems that AFB1 can induce cytotoxicity and necrosis in bovine mammary epithelial cells. PMID:24312827

  11. Non-Neuronal Cells Are Required to Mediate the Effects of Neuroinflammation: Results from a Neuron-Enriched Culture System

    PubMed Central

    Hui, Chin Wai; Zhang, Yang; Herrup, Karl

    2016-01-01

    Chronic inflammation is associated with activated microglia and reactive astrocytes and plays an important role in the pathogenesis of neurodegenerative diseases such as Alzheimer’s. Both in vivo and in vitro studies have demonstrated that inflammatory cytokine responses to immune challenges contribute to neuronal death during neurodegeneration. In order to investigate the role of glial cells in this phenomenon, we developed a modified method to remove the non-neuronal cells in primary cultures of E16.5 mouse cortex. We modified previously reported methods as we found that a brief treatment with the thymidine analog, 5-fluorodeoxyuridine (FdU), is sufficient to substantially deplete dividing non-neuronal cells in primary cultures. Cell cycle and glial markers confirm the loss of ~99% of all microglia, astrocytes and oligodendrocyte precursor cells (OPCs). More importantly, under this milder treatment, the neurons suffered neither cell loss nor any morphological defects up to 2.5 weeks later; both pre- and post-synaptic markers were retained. Further, neurons in FdU-treated cultures remained responsive to excitotoxicity induced by glutamate application. The immunobiology of the FdU culture, however, was significantly changed. Compared with mixed culture, the protein levels of NFκB p65 and the gene expression of several cytokine receptors were altered. Individual cytokines or conditioned medium from β-amyloid-stimulated THP-1 cells that were, potent neurotoxins in normal, mixed cultures, were virtually inactive in the absence of glial cells. The results highlight the importance of our glial-depleted culture system and identifies and offer unexpected insights into the complexity of -brain neuroinflammation. PMID:26788729

  12. Assessing the immunomodulatory role of heteroglycan in a tumor spheroid and macrophage co-culture model system.

    PubMed

    Devi, K Sanjana P; Mishra, Debasish; Roy, Bibhas; Ghosh, Sudip K; Maiti, Tapas K

    2015-01-01

    The therapeutic benefits of glycans have garnered much attention over the last few decades with most studies being reported in 2D cultures or in animal models. The present work is therefore aimed to assess the effects of an immunomodulatory heteroglycan in a 3D milieu. Briefly, HT29 tumor spheroids were incubated with THP-1 macrophages at 1:1 ratio in a culture medium supplemented with immune stimulants such as heteroglycans or LPS. Spheroidal distortion, migration of tumor cells from the periphery of the spheroids and 46% of higher macrophage invasiveness was noted in heteroglycan-treated co-cultures with respect to control cultures. Histological sections of the treated co-cultures revealed the presence of high apoptotic tumor cells in the spheroidal periphery. CD11c and CD68 staining further suggested the predominance of macrophages in the vicinity of the apoptotic tumor cells. Such an in vitro created tissue system may thereby confirm the effectiveness of heteroglycan in activating the immune cells to exhibit tumor cytotoxic properties. PMID:25965450

  13. Amphibian (Euphlyctis cyanophlyctis) in vitro ovarian culture system to assess impact of aquatic agrochemical contaminants on female reproduction.

    PubMed

    Katti, Pancharatna A; Ghodgeri, Manjunath G; Goundadkar, Basavaraj B

    2016-01-01

    The present study is an attempt to screen impacts of aquatic agrochemical contaminants (acephate, atrazine and cypermethrin) on development and growth of follicles, in in vitro-cultured ovarian fragments of frog (Euphlyctis cyanophlyctis). Ovarian lobes removed surgically from gravid females were cut into small pieces and cultured in vitro in presence of graded (0.01 or 0.1 μg/ml of culture medium) concentrations of test chemicals or estradiol-17β (positive controls) or culture medium alone (controls) in quadruplicate sets at 23 ± 1 °C temperature for 20 days in a humidified sterile chamber. On 21st day, they were fixed in Bouin's fluid and used for differential follicle counting (n = 3 sets) and histology (n = 1 set). In vitro exposure of ovarian fragments to test chemicals caused a decline in previtellogenic follicles, maintenance of large yolky follicles, incorporation of brown granules into early vitellogenic follicles and decrease in follicular atresia compared to corresponding controls. These results suggest that ovarian follicles are greatly sensitive to chemical exposure during their transition from previtellogenic to vitellogenic growth phase and in vitro ovarian culture system may be used as a tool to assess the effects of aquatic agrochemical contaminants on ovarian function. PMID:25945413

  14. Establishment of feeder-free culture system for human induced pluripotent stem cell on DAS nanocrystalline graphene

    PubMed Central

    Lee, Hyunah; Nam, Donggyu; Choi, Jae-Kyung; Araúzo-Bravo, Marcos J.; Kwon, Soon-Yong; Zaehres, Holm; Lee, Taehee; Park, Chan Young; Kang, Hyun-Wook; Schöler, Hans R.; Kim, Jeong Beom

    2016-01-01

    The maintenance of undifferentiated human pluripotent stem cells (hPSC) under xeno-free condition requires the use of human feeder cells or extracellular matrix (ECM) coating. However, human-derived sources may cause human pathogen contamination by viral or non-viral agents to the patients. Here we demonstra