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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. Bacterial culture detection and identification in blood agar plates with an optoelectronic nose.

    PubMed

    Lim, Sung H; Mix, Samantha; Anikst, Victoria; Budvytiene, Indre; Eiden, Michael; Churi, Yair; Queralto, Nuria; Berliner, Anders; Martino, Raymond A; Rhodes, Paul A; Banaei, Niaz

    2016-02-01

    Clinical microbiology automation is currently limited by the lack of an in-plate culture identification system. Using an inexpensive, printed, disposable colorimetric sensor array (CSA) responsive to the volatiles emitted into plate headspace by microorganisms during growth, we report here that not only the presence but the species of bacteria growing in plate was identified before colonies are visible. In 1894 trials, 15 pathogenic bacterial species cultured on blood agar were identified with 91.0% sensitivity and 99.4% specificity within 3 hours of detection. The results indicate CSAs integrated into Petri dish lids present a novel paradigm to speciate microorganisms, well-suited to integration into automated plate handling systems.

  5. 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

  6. Rapid Identification of Positive Blood Cultures by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Using Prewarmed Agar Plates

    PubMed Central

    Bhatti, M. M.; Boonlayangoor, S.; Beavis, K. G.

    2014-01-01

    This study describes an inexpensive and straightforward method for identifying bacteria by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) directly from positive blood cultures using prewarmed agar plates. Different inoculation methods and incubation times were evaluated to determine the optimal conditions. The two methods using pelleted material from positive culture bottles performed best. In particular, the pellet streak method correctly identified 94% of the Gram negatives following 4 h of incubation and 98% of the Gram positives following 6 h of incubation. PMID:25232166

  7. Rapid identification of positive blood cultures by matrix-assisted laser desorption ionization-time of flight mass spectrometry using prewarmed agar plates.

    PubMed

    Bhatti, M M; Boonlayangoor, S; Beavis, K G; Tesic, V

    2014-12-01

    This study describes an inexpensive and straightforward method for identifying bacteria by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) directly from positive blood cultures using prewarmed agar plates. Different inoculation methods and incubation times were evaluated to determine the optimal conditions. The two methods using pelleted material from positive culture bottles performed best. In particular, the pellet streak method correctly identified 94% of the Gram negatives following 4 h of incubation and 98% of the Gram positives following 6 h of incubation.

  8. 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

  9. Rectal screening for Klebsiella pneumoniae carbapenemases: comparison of real-time PCR and culture using two selective screening agar plates.

    PubMed

    Singh, Kamaljit; Mangold, Kathy A; Wyant, Kody; Schora, Donna M; Voss, Barbara; Kaul, Karen L; Hayden, Mary K; Chundi, Vishnu; Peterson, Lance R

    2012-08-01

    Klebsiella pneumoniae carbapenemases (KPCs) have recently been described in Chicago, IL, especially among residents of long-term acute care hospitals (LTACHs). These patients are frequently transferred to local Chicago hospitals for higher acuity of medical care, and rapid detection and isolation of KPC-colonized LTACH residents may interrupt the introduction of KPCs into acute care hospitals. We evaluated the performance of a real-time PCR for bla(KPC) from enrichment broth versus direct plating of rectal surveillance swabs on two selective culture media, CHROMagar extended-spectrum-β-lactamase (ESBL) and vancomycin, amphotericin B, ceftazidime, and clindamycin (VACC) plates. Rectal surveillance swabs were collected as part of a point prevalence study of KPC carriage rates among 95 residents of two Chicago area LTACHs. Discrepant results between PCR and culture were resolved by subculturing the enrichment broth. Overall, 66 of 95 patients (69.5%) were colonized with KPCs, using the cumulative results of culture as a reference standard. Real-time PCR from enrichment broth was positive in 64 of 66 (97%) colonized patients, including nine surveillance swabs that were missed by both selective culture media. PCR demonstrated higher sensitivity, 97.0%, than culture using either CHROMagar or VACC plates (both with sensitivity of 77.3%). In addition, turnaround time was significantly shorter for the PCR-based method than for culture, with a mean of 24 h versus 64 to 72 h for CHROMagar and VACC plates (P < 0.0001). Overall, PCR for bla(KPC) represents the best screening test for KPCs with significantly higher sensitivity and with less hands-on time, resulting in a shorter time to results.

  10. A combined approach for the enhanced detection and isolation of Bartonella species in dog blood samples: pre-enrichment liquid culture followed by PCR and subculture onto agar plates.

    PubMed

    Duncan, Ashlee W; Maggi, Ricardo G; Breitschwerdt, Edward B

    2007-05-01

    Historically, direct plating, lysis centrifugation, or freeze-thaw approaches have proven to be highly insensitive methods for confirming Bartonella species infection in dogs. A prospective study was designed to compare diagnostic methods for the detection of Bartonella using samples submitted to the Vector-Borne Disease Diagnostic Laboratory at North Carolina State University. Methods included indirect immunofluorescence assay, PCR, direct inoculation of a blood agar plate (trypticase soy agar with 5% rabbit blood), and inoculation into a novel pre-enrichment liquid medium, Bartonella/alpha-Proteobacteria growth medium (BAPGM). Sequential research efforts resulted in the development of a combinational approach consisting of pre-enrichment culture of Bartonella species in BAPGM, sub-inoculation of the liquid culture onto agar plates, followed by DNA amplification using PCR. The multi-faceted approach resulted in substantial improvement in the microbiological detection and isolation of Bartonella when compared to direct inoculation of a blood agar plate. Importantly, this approach facilitated the detection and subsequent isolation of both single and co-infections with two Bartonella species in the blood of naturally infected dogs. The use of a combinational approach of pre-enrichment culture and PCR may assist in the diagnostic confirmation of bartonellosis in dogs and other animals.

  11. Automated counting of bacterial colony forming units on agar plates.

    PubMed

    Brugger, Silvio D; Baumberger, Christian; Jost, Marcel; Jenni, Werner; Brugger, Urs; Mühlemann, Kathrin

    2012-01-01

    Manual counting of bacterial colony forming units (CFUs) on agar plates is laborious and error-prone. We therefore implemented a colony counting system with a novel segmentation algorithm to discriminate bacterial colonies from blood and other agar plates.A colony counter hardware was designed and a novel segmentation algorithm was written in MATLAB. In brief, pre-processing with Top-Hat-filtering to obtain a uniform background was followed by the segmentation step, during which the colony images were extracted from the blood agar and individual colonies were separated. A Bayes classifier was then applied to count the final number of bacterial colonies as some of the colonies could still be concatenated to form larger groups. To assess accuracy and performance of the colony counter, we tested automated colony counting of different agar plates with known CFU numbers of S. pneumoniae, P. aeruginosa and M. catarrhalis and showed excellent performance.

  12. Evaluation of an automated agar plate streaker.

    PubMed Central

    Tilton, R C; Ryan, R W

    1978-01-01

    An automated agar plate streaker was evaluated. The Autostreaker mechanizes the agar plate streaking process by providing storage for plates, labeling and streaking one or more plates for either isolation or quantitation, and stacking in one of several racks for subsequent incubation. Results showed the Autostreaker to produce agar plates with well-separated colonies and accurate colony counts. A total of 1,930 clinical specimens were processed either in parallel with manual methods or solely by the Autostreaker. Technologist acceptance of machine-streaked plates was outstanding. Images PMID:348722

  13. Syneresis and delayed detachment in agar plates

    NASA Astrophysics Data System (ADS)

    Divoux, Thibaut; Mao, Bosi; Snabre, Patrick

    Biogels made of crosslinked polymers such as proteins or polysaccharides behave as porous soft solids and store large amount of solvent. These gels undergo spontaneous aging, called syneresis that consists in the shrinkage of the gel matrix and the progressive expulsion of the solvent. As a result, a biogel originally casted in a container often lose contact with the container sidewalls, and the detachment time is a priori difficult to anticipate since it may occur over variable time spans (from hours to days). Here we report on the syneresis phenomena in agar plates that consist in 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 $e_{min}$ measured along the sidewall of the plate, and increases as a robust function of $e_{min}$ 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 the 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.

  14. 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.

  15. 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

  16. 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 §...

  17. 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 §...

  18. 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 §...

  19. 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 §...

  20. 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 §...

  1. A modified method for the detection of microbial proteases on agar plates using tannic acid.

    PubMed

    Saran, Saurabh; Isar, Jasmine; Saxena, Rajendra Kumar

    2007-06-10

    In routine assay for the screening of microbes producing proteases, 10% trichloroaceticacid (TCA) is flooded on the milk agar plates after inoculation and required incubation to precipitate the protein. However, the clarity of the hydrolyzed zone is not very sharp and distinct. We herein present an improved assay for detecting the presence of extracellular protease from microorganisms on agar plates. In this method 10% tannic acid is flooded on the milk agar plate (in place of, TCA) to observe the zone of hydrolysis. Tannic acid sharply increases the colour intensity of the plate, as it favours the precipitation of the unhydrolyzed protein in the plate, thereby improving the contrast between the intact zones and the enzymatic lyses zones of the substrate. Our results indicate that this method is useful to detect extracellular proteases produced by both fungi as well as bacteria. The method used in the present study is sensitive, and can be easily performed for screening of large number of microbial cultures. This is the first report on the use of tannic acid for the detection of microbial proteases.

  2. 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

  3. Solvent-tolerance of fungi located on an interface between an agar plate and an organic solvent.

    PubMed

    Oda, Shinobu; Sugitani, Ayaka; Ohashi, Shinichi

    2014-01-01

    While 6 by 20 of type culture fungi could grow on an interface between organic solvent (log P, 4.12) and agar plate, 13 by 20 of strains could form a large colony after the removal of more toxic solvent, such as styrene (log P, 2.95) and tert-butyl acetate (log P, 1.76) because of viability of spores on the interface.

  4. 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 Central

    Ingram, David T.; Lamichhane, Chinta M.; Rollins, David M.; Carr, Lewis E.; Mallinson, Edward T.; Joseph, Sam W.

    1998-01-01

    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°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 traditional

  5. Ceftibuten-containing agar plate for detecting group B streptococci with reduced penicillin susceptibility (PRGBS).

    PubMed

    Kamiya, Chitose; Kimura, Kouji; Doyama, Yo; Miyazaki, Akira; Morimoto, Makiko; Banno, Hirotsugu; Nagano, Noriyuki; Jin, Wanchun; Wachino, Jun-ichi; Yamada, Keiko; Arakawa, Yoshichika

    2015-08-01

    Penicillins remain first-line agents for treatment of group B Streptococcus (Streptococcus agalactiae; GBS) infections; however, several reports have confirmed the existence of GBS with reduced penicillin susceptibility (PRGBS). Because no selective agar plates for detection of PRGBS are available to date, in this investigation, we developed the selective agar plate for detection of PRGBS. We used 19 genetically well-confirmed PRGBS isolates and 38 penicillin-susceptible GBS isolates identified in Japan. For preparation of trial PRGBS-selective agar plates, we added 1 of antimicrobial agents (among oxacillin, ceftizoxime, and ceftibuten) to a well-established GBS-selective agar plate. Among 12 trial PRGBS-selective agar plates, Muller-Hinton agar containing 128 μg/mL ceftibuten with 5% sheep blood, 8 μg/mL gentamicin, and 12 μg/mL nalidixic acid was the most appropriate selective agar for PRGBS, showing 100% sensitivity and 81.6% specificity. In cases of potential nosocomial spread of PRGBS, the selective agar plate could be useful and reliable.

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

    PubMed

    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

    2012-09-01

    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.

  7. 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.

  8. 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.

  9. 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

  10. 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.

  11. Thallium toxicosis in a dog consequent to ingestion of Mycoplasma agar plates.

    PubMed

    Puschner, Birgit; Basso, Marguerite M; Graham, Thomas W

    2012-01-01

    A 1-year-old dog ingested a mixture of blood agar and Mycoplasma agar plates. The Mycoplasma agar plates contained thallium acetate, which resulted in an estimated minimum dose of 5 mg thallium acetate/kg bodyweight. Clinical signs over the course of 2-3 weeks included vomiting, diarrhea, weight loss, alopecia, dysphonia, ataxia, paresthesia, intension tremors, megaesophagus with subsequent aspiration pneumonia, and several seizure episodes. The dog was treated with intravenous fluids and placement of a gastric feeding tube. Thallium concentrations in hair were 8.2 µg/g in samples taken on day 19, 16.4 µg/g in samples taken 3 months after exposure, 13.4 µg/g in samples taken 5 months after exposure, and nondetectable in samples taken 7 months after exposure. The blood thallium concentration was 190 µg/l on day 19 and nondetec table 3 months after exposure. Megaesophagus and dysphonia continued for 10 months after exposure. This case of thallium poisoning following ingestion of mycoplasma agar plates demonstrates that unusual sources of thallium still exist and suggests that thallium toxicosis should be included in the list of differential diagnoses in dogs presented with megaesophagus, especially if alopecia and other unexplained peripheral neuropathies are present. Hair and blood samples are useful specimens to reach an accurate diagnosis even if taken several weeks post exposure. The postexposure blood and hair thallium concentrations reported in this case are useful data for diagnosticians investigating dogs with potential thallium poisoning.

  12. Evaluation of Granada agar plate for detection of Streptococcus agalactiae in urine specimens from pregnant women.

    PubMed

    Tamayo, Javier; Gómez-Garcés, José-Luis; Alós, Juan-Ignacio

    2004-08-01

    The Granada agar plate (GAP; Biomedics SL, Madrid, Spain) was evaluated for the detection of group B streptococci (GBS) in urine specimens from pregnant women submitted for testing for asymptomatic bacteriuria and was compared with blood agar (BA [Columbia agar with 5% sheep blood]; bioMérieux, Marcy l'Etoile, France). The GAP detected 103 out of 105 GBS, whereas BA detected only 50. Use of the GAP could be a good method for the detection of GBS in urine specimens from pregnant women. PMID:15297542

  13. 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

  14. 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

  15. Characterization of multidrug-resistant group B streptococci with reduced penicillin susceptibility forming small non-Beta-hemolytic colonies on sheep blood agar plates.

    PubMed

    Banno, Hirotsugu; Kimura, Kouji; Tanaka, Yosuke; Kitanaka, Hiromitsu; Jin, Wanchun; Wachino, Jun-ichi; Yamada, Keiko; Shibayama, Keigo; Arakawa, Yoshichika

    2014-06-01

    We isolated and characterized three multidrug-resistant clinical isolates of group B streptococci with reduced penicillin susceptibility (PRGBS) that formed small non-beta-hemolytic colonies on sheep blood agar plates but grew well on chocolate agar plates. They can be overlooked in the bacterial identification step, leading to clinical misdiagnosis and treatment failure.

  16. 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

  17. Evaluation of an immunochromatographic assay for direct identification of thermostable direct hemolysin-producing Vibrio parahaemolyticus colonies on selective agar plates.

    PubMed

    Kawatsu, Kentaro; Sakata, Junko; Yonekita, Taro; Kumeda, Yuko

    2015-12-01

    We evaluated the utility of an immunochromatographic assay (NH IC TDH) in identifying thermostable direct hemolysin (TDH)-producing Vibrio parahaemolyticus colonies on selective agar plates. The sensitivity of the NH IC TDH assay was 100% (189 samples) and its specificity was 100% (41 samples) compared with the presence of tdh.

  18. 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

  19. Evaluation of Petrifilm™ aerobic count plates as an equivalent alternative to drop plating on R2A agar plates in a biofilm disinfectant efficacy test.

    PubMed

    Fritz, B G; Walker, D K; Goveia, D E; Parker, A E; Goeres, D M

    2015-03-01

    This paper compares Petrifilm™ aerobic count (AC) plates to drop plating on R2A agar plates as an alternative method for biofilm bacteria enumeration after application of a disinfectant. A Pseudomonas aeruginosa biofilm was grown in a Centers for Disease Control and Prevention biofilm reactor (ASTM E2562) and treated with 123 ppm sodium hypochlorite (as free chlorine) according to the Single Tube Method (ASTM E2871). Aliquots from the same dilution tubes were plated on Petrifilm™ AC plates and drop plated on R2A agar plates. The Petrifilm™ AC and R2A plates were incubated for 48 and 24 h, respectively, at 36 ± 1 °C. After nine experimental runs performed by two technicians, the mean difference in biofilm log densities [log biofilm density (LD) = log10(CFU/cm(2))] between the two methods for control coupons, treated coupons, and log reduction (LR) was 0.052 (p = 0.451), -0.102 (p = 0.303), and 0.152 (p = 0.313). Equivalence testing was used to assess equivalence of the two plating methods. The 90 % confidence intervals for the difference in control and treated mean LDs between methods were (-0.065, 0.170) and (-0.270, 0.064), both of which fall within a (-0.5, +0.5) equivalence criterion. The 90 % confidence interval for the mean LR difference (-0.113, 0.420) also falls within this equivalence criterion. Thus, Petrifilm™ AC plates were shown to be statistically equivalent to drop plating on R2A agar for the determination of control LDs, treated LDs, and LR values in an anti-biofilm efficacy test. These are the first published results that establish equivalency to a traditional plate counting technique for biofilms and for a disinfectant assay.

  20. Gene escape model: Transfer of heavy metal resistance genes from Escherichia coli to Alcaligenes eutrophus on agar plates and in soil samples

    SciTech Connect

    Top, E. Studiecentrum voor Kernenergie, Mol ); Mergeay, M.; Springael, D. ); Verstraete, W. )

    1990-08-01

    Conjugal transfer from Escherichia coli to Alcaligenes eutrophus of the A. eutrophus genes coding for plasmid-borne resistance to cadmium, cobalt, and zinc (czc genes) was investigated on agar plates and in soil samples. This czc fragment is not expressed in the donor strain, E. coli, but it is expressed in the recipient strain, A. eutrophus. Hence, expression of heavy metal resistance by cells plated on a medium containing heavy metals represents escape of the czc genes. The two plasmids into which this DNA fragment has been cloned previously and which were used in these experiments are the nonconjugative, mobilizable plasmid pDN705 and the nonconjugative, nonmobilizable plasmid pMOL149. The results demonstrate that even genes incorporated into nonmobilizable plasmids can be exchanged between two different genera and that the presence of broad-host-range plasmids in putative recipients among soil bacteria could increase the risk of gene dissemination in case of release of genetically engineered microorganisms. The results also reveal that in certain soils, environmental conditions and particularly nutrient levels are conducive to gene transfer.

  1. Wood and humus decay strategies by white-rot basidiomycetes correlate with two different dye decolorization and enzyme secretion patterns on agar plates.

    PubMed

    Barrasa, José M; Blanco, María N; Esteve-Raventós, Fernando; Altés, Alberto; Checa, Julia; Martínez, Angel T; Ruiz-Dueñas, Francisco J

    2014-11-01

    During several forays for ligninolytic fungi in different Spanish native forests, 35 white-rot basidiomycetes growing on dead wood (16 species from 12 genera) and leaf litter (19 species from 10 genera) were selected for their ability to decolorize two recalcitrant aromatic dyes (Reactive Blue 38 and Reactive Black 5) added to malt extract agar medium. In this study, two dye decolorization patterns were observed and correlated with two ecophysiological groups (wood and humus white-rot basidiomycetes) and three taxonomical groups (orders Polyporales, Hymenochaetales and Agaricales). Depending on the above groups, different decolorization zones were observed on the dye-containing plates, being restricted to the colony area or extending to the surrounding medium, which suggested two different decay strategies. These two strategies were related to the ability to secrete peroxidases and laccases inside (white-rot wood Polyporales, Hymenochaetales and Agaricales) and outside (white-rot humus Agaricales) of the fungal colony, as revealed by enzymatic tests performed directly on the agar plates. Similar oxidoreductases production patterns were observed when fungi were grown in the absence of dyes, although the set of enzyme released was different. All these results suggest that the decolorization patterns observed could be related with the existence of two decay strategies developed by white-rot basidiomycetes adapted to wood and leaf litter decay in the field.

  2. 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.

  3. Culture systems: air quality.

    PubMed

    Thomas, Theodore

    2012-01-01

    Poor laboratory air quality is a known hazard to the culture of human gametes and embryos. Embryologists and chemists have employed analytical methods for identifying and measuring bulk and select air pollutants to assess the risk they pose to the embryo culture system. However, contaminant concentrations that result in gamete or embryotoxicity are poorly defined. Combating the ill effects of poor air quality requires an understanding of how toxicants can infiltrate the laboratory, the incubator, and ultimately the culture media. A further understanding of site-specific air quality can then lead to the consideration of laboratory design and management strategies that can minimize the deleterious effects that air contamination may have on early embryonic development in vitro.

  4. 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.

  5. Culture and Social Systems Revisited

    ERIC Educational Resources Information Center

    Parsons, Talcott

    1972-01-01

    A brief statement of the history of the relation between culture concepts is given, then, proceeding on the assumption that all human societies are interpenetrated with culture, the author attempts a relatively systematic outline of the structure of cultural systems and of their modes of articulation in social systems. (JB)

  6. 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.

  7. 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.

  8. 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.

  9. Culture systems: embryo density.

    PubMed

    Reed, Michael L

    2012-01-01

    Embryo density is defined as the embryo-to-volume ratio achieved during in vitro culture; in other words, it is the number of embryos in a defined volume of culture medium. The same density can be achieved by manipulating either the number of embryos in a given volume of medium, or manipulating the volume of the medium for a given number of embryos: for example, a microdrop with five embryos in a 50 μl volume under oil has the same embryo-to-volume ratio (1:10 μl) as a microdrop with one embryo in a 10 μl volume under oil (1:10 μl). Increased embryo density can improve mammalian embryo development in vitro; however, the mechanism(s) responsible for this effect may be different with respect to which method is used to increase embryo density.Standard, flat sterile plastic petri dishes are the most common, traditional platform for embryo culture. Microdrops under a mineral oil overlay can be prepared to control embryo density, but it is critical that dish preparation is consistent, where appropriate techniques are applied to prevent microdrop dehydration during preparation, and results of any data collection are reliable, and repeatable. There are newer dishes available from several manufacturers that are specifically designed for embryo culture; most are readily available for use with human embryos. The concept behind these newer dishes relies on fabrication of conical and smaller volume wells into the dish design, so that embryos rest at the lowest point in the wells, and where putative embryotrophic factors may concentrate.Embryo density is not usually considered by the embryologist as a technique in and of itself; rather, the decision to culture embryos in groups or individually is protocol-driven, and is based more on convenience or the need to collect data on individual embryos. Embryo density can be controlled, and as such, it can be utilized as a simple, yet effective tool to improve in vitro development of human embryos. PMID:22829380

  10. Culture systems: low-oxygen culture.

    PubMed

    Kovačič, Borut

    2012-01-01

    The tension of oxygen measured in the oviducts of several mammals was 5-8.7 %, but this drops in the uterine milieu to <2 % in cows and primates. For embryo culture in human in vitro fertilization (IVF), a non-physiologic level of 20 % oxygen has been used for the past 30 years. However, several animal studies have shown that low levels of oxygen plays an important physiological role in reducing the high levels of detrimental reactive oxygen species within cells, influences the embryonic gene expression, helps with embryo metabolism of glucose, and enhances embryo development to blastocysts. However, clinical studies have given contradictory results. Nevertheless, in nearly all reports, some kind of improvement has been observed, either in embryo development or in implantation and no detriments have been reported. For these reasons, more and more IVF laboratories utilize low oxygen during embryo culture.

  11. Culture systems: embryo culture and monozygotic twinning.

    PubMed

    Sparks, Amy E

    2012-01-01

    The incidence of monozygotic twinning in pregnancies achieved with assisted reproductive technologies (ART) is significantly higher than spontaneously conceived pregnancies. The factors associated with ART that predispose the embryos to splitting are not well-characterized. Assisted hatching and extended embryo culture are two ART laboratory methods that have been risk factors for monozygotic twinning. The methods and strategies that may be employed to avoid monozygotic twinning are discussed in this chapter.

  12. Stability of repetitive-sequence PCR patterns with respect to culture age and subculture frequency.

    PubMed

    Kang, Hyunseok Peter; Dunne, W Michael

    2003-06-01

    To examine the stability of repetitive-sequence (rep) PCR profiles, six species of bacteria were subcultured to blood agar plates and DNA was extracted from the cultures after 24, 48, and 72 h of incubation at 35 degrees C. In addition, the same species were subcultured to fresh blood plates daily and DNA was extracted from the cultures after growth of 5, 10, and 15 subcultures, respectively. rep PCR analysis demonstrated that all rep PCR fingerprints from a single species were identical.

  13. 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

  14. 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

  15. The rich detail of cultural symbol systems.

    PubMed

    Read, Dwight W

    2014-08-01

    The goal of forming a science of intentional behavior requires a more richly detailed account of symbolic systems than is assumed by the authors. Cultural systems are not simply the equivalent in the ideational domain of culture of the purported Baldwin Effect in the genetic domain. PMID:25162879

  16. Evaluation of Bio-Rad MRSASelect agar for detection of methicillin-resistant Staphylococcus aureus directly from blood cultures.

    PubMed

    Riedel, Stefan; Dam, Lisa; Stamper, Paul D; Shah, Syed A R; Carroll, Karen C

    2010-06-01

    MRSASelect agar (Bio-Rad, Redmond, WA) was evaluated for its performance in detecting MRSA directly from positive blood cultures containing Gram-positive cocci in clusters. Agar plates were evaluated for the presence of pink colonies at 18 to 24 h. Results were compared to organism identification by using standard laboratory methods. Confirming coagulase on pink isolates, the sensitivity and specificity were both 99%.

  17. Nested culture method improves detection of Fusobacterium from stool in patients with ulcerative colitis.

    PubMed

    Yukawa, Toyokazu; Ohkusa, Toshifumi; Shibuya, Tomoyoshi; Tsukinaga, Shintaro; Mitobe, Jimi; Takakura, Kazuki; Takahara, Akitaka; Odahara, Shunichi; Matsudaira, Hiroshi; Nagatsuma, Keisuke; Kitahara, Takuya; Kajihara, Mikio; Uchiyama, Kan; Arakawa, Hiroshi; Koido, Shigeo; Tajiri, Hisao

    2013-01-01

    Fusobacterium varium is an elusive pathogenic factor in ulcerative colitis (UC); conventional methods of fecal culture rarely recover F. varium. We have developed a nested culture method to recover Fusobacterium and we used it to investigate whether F. varium could be isolated from UC patients. We enrolled 50 consecutive patients in this study; 26 received combination antibiotic therapy that included amoxicillin, tetracycline, and metronidazole (ATM) for 2 weeks and were thus assigned to the ATM group, and the remaining 24 were assigned to the non-ATM group and did not receive any antibiotics. Stool samples were added to 10 ml of GAM broth that contained neomycin and crystal violet. The samples were vortexed and incubated under anaerobic conditions. The preincubated broth was streaked onto a Fusobacterium-selective agar plate and then incubated under anaerobic conditions. The species of the colonies isolated were identified using the Vitek Automated system and PCR analysis. We recoverd F. varium from 7 of the 24 non-ATM patients (29.2%) and none from the ATM patients (0%) (P = 0.0035). All of the F. varium isolates were susceptible to ATM. This study suggests that the recovery of F. varium is related to UC, which aligns with results from previous studies that used mucosal culture, immunostaining, real-time PCR, and serological studies.

  18. 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…

  19. Detection of bacterioplankton in immersed cadavers using selective agar plates.

    PubMed

    Kakizaki, Eiji; Kozawa, Shuji; Tashiro, Noriko; Sakai, Masahiro; Yukawa, Nobuhiro

    2009-04-01

    We measured bacterioplankton in blood from cadavers retrieved from the sea (n=12), near estuaries (n=4), rivers (fresh water, n=8) and from bathtubs (n=4) as well as from non-drowned victims (n=10) discovered near aquatic environments. Blood from 11 victims drowned in seawater developed bioluminescent and/or blue colonies (oxidase test positive) on selective media containing 2-4% NaCl. Homology analyses of the 16S rRNA gene showed that all of them were marine bacteria (genera: Photobacterium, Vibrio, Shewanella, Psychrobacter). Blood from all victims drowned in rivers generated blue colonies on plates containing 3%, but not 4% NaCl. Homology analyses showed that the blue colonies were generated from bacteria that inhabit fresh water (Aeromonas). None of the blood samples from victims that drowned in bathtubs generated bioluminescent and blue colonies. However, all cadavers contained bacteria that produced unstained colonies (Staphylococcus, Bacillus, Enterobacter, Escherichia, etc.). Among non-drowned victims, blood from two gave rise to blue colonies on plates containing < or =3% NaCl (Pseudomonas). Of the cadavers found near estuaries, bioluminescent and blue colonies developed from two of them on media containing 2-4% NaCl (Photobacterium, Vibrio, Listonella), but not from two others on plates containing 4% NaCl (at < or =3%; blue colonies, Aeromonas; unstained colonies, Citrobacter, Vagococcus, Proteus, Enterobacter). These results suggested that the presence of numerous bacterioplankton in immersed cadavers could support a conclusion of death by drowning.

  20. 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

  1. 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.

  2. 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

  3. 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.

  4. 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

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

    NASA Astrophysics Data System (ADS)

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

    2016-09-01

    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.

  6. 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

  7. Preparation of a blood culture pellet for rapid bacterial identification and antibiotic susceptibility testing.

    PubMed

    Croxatto, Antony; Prod'hom, Guy; Durussel, Christian; Greub, Gilbert

    2014-10-15

    Bloodstream infections and sepsis are a major cause of morbidity and mortality. The successful outcome of patients suffering from bacteremia depends on a rapid identification of the infectious agent to guide optimal antibiotic treatment. The analysis of Gram stains from positive blood culture can be rapidly conducted and already significantly impact the antibiotic regimen. However, the accurate identification of the infectious agent is still required to establish the optimal targeted treatment. We present here a simple and fast bacterial pellet preparation from a positive blood culture that can be used as a sample for several essential downstream applications such as identification by MALDI-TOF MS, antibiotic susceptibility testing (AST) by disc diffusion assay or automated AST systems and by automated PCR-based diagnostic testing. The performance of these different identification and AST systems applied directly on the blood culture bacterial pellets is very similar to the performance normally obtained from isolated colonies grown on agar plates. Compared to conventional approaches, the rapid acquisition of a bacterial pellet significantly reduces the time to report both identification and AST. Thus, following blood culture positivity, identification by MALDI-TOF can be reported within less than 1 hr whereas results of AST by automated AST systems or disc diffusion assays within 8 to 18 hr, respectively. Similarly, the results of a rapid PCR-based assay can be communicated to the clinicians less than 2 hr following the report of a bacteremia. Together, these results demonstrate that the rapid preparation of a blood culture bacterial pellet has a significant impact on the identification and AST turnaround time and thus on the successful outcome of patients suffering from bloodstream infections.

  8. Background culturable bacteria aerosol in two large public buildings using HVAC filters as long term, passive, high-volume air samplers.

    PubMed

    Stanley, Nicholas J; Kuehn, Thomas H; Kim, Seung Won; Raynor, Peter C; Anantharaman, Senthilvelan; Ramakrishnan, M A; Goyal, Sagar M

    2008-04-01

    Background culturable bacteria aerosols were collected and identified in two large public buildings located in Minneapolis, Minnesota and Seattle, Washington over a period of 5 months and 3 months, respectively. The installed particulate air filters in the ventilation systems were used as the aerosol sampling devices at each location. Both pre and final filters were collected from four air handing units at each site to determine the influence of location within the building, time of year, geographical location and difference between indoor and outdoor air. Sections of each loaded filter were eluted with 10 ml of phosphate buffered saline (PBS). The resulting solutions were cultured on blood agar plates and incubated for 24 h at 36 degrees C. Various types of growth media were then used for subculturing, followed by categorization using a BioLog MicroStation (Biolog, Hayward, CA, USA) and manual observation. Environmental parameters were gathered near each filter by the embedded on-site environmental monitoring systems to determine the effect of temperature, humidity and air flow. Thirty nine different species of bacteria were identified, 17 found only in Minneapolis and 5 only in Seattle. The hardy spore-forming genus Bacillus was the most commonly identified and showed the highest concentrations. A significant decrease in the number of species and their concentration occurred in the Minneapolis air handling unit supplying 100% outdoor air in winter, however no significant correlations between bacteria concentration and environmental parameters were found.

  9. Automatic Digital Plate Reading for Surveillance Cultures.

    PubMed

    Kirn, Thomas J

    2016-10-01

    The automation of specimen processing and culture workup has rapidly emerged in clinical microbiology laboratories throughout the world and more recently in the United States. While many U.S. laboratories have implemented some form of automated specimen processing and some have begun performing digital plate reading, automated colony analysis is just beginning to be utilized clinically. In this issue of the Journal of Clinical Microbiology, M. L. Faron et al. (J Clin Microbiol 54:2470-2475, 2016, http://dx.doi.org/10.1128/JCM.01040-16) report the results of their evaluation of the performance of the WASPLab Chromogenic Detection Module (CDM) for categorizing chromogenic agar plates as negative or "nonnegative" for vancomycin-resistant enterococci (VRE). Their major finding was 100% sensitivity for detection of "nonnegative" specimens using CDM compared to manual methods for specimens plated on two different types of VRE chromogenic agar plates. Additionally, utilization of digital plate reading in conjunction with automated colony analysis was predicted to result in significant savings based on greatly reduced labor costs.

  10. 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

  11. 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.

  12. 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

  13. Family systems theory, attachment theory, and culture.

    PubMed

    Rothbaum, Fred; Rosen, Karen; Ujiie, Tatsuo; Uchida, Nobuko

    2002-01-01

    Family systems theory and attachment theory have important similarities and complementarities. Here we consider two areas in which the theories converge: (a) in family system theorists' description of an overly close, or "enmeshed," mother-child dyad, which attachment theorists conceptualize as the interaction of children's ambivalent attachment and mothers' preoccupied attachment; (b) in family system theorists' description of the "pursuer-distance cycle" of marital conflict, which attachment theorists conceptualize as the interaction of preoccupied and dismissive partners. We briefly review family systems theory evidence, and more extensively review attachment theory evidence, pertaining to these points of convergence. We also review cross-cultural research, which leads us to conclude that the dynamics described in both theories reflect, in part, Western ways of thinking and Western patterns of relatedness. Evidence from Japan suggests that extremely close ties between mother and child are perceived as adaptive, and are more common, and that children experience less adverse effects from such relationships than do children in the West. Moreover, in Japan there is less emphasis on the importance of the exclusive spousal relationship, and less need for the mother and father to find time alone to rekindle romantic, intimate feelings and to resolve conflicts by openly communicating their differences. Thus, the "maladaptive" pattern frequently cited by Western theorists of an extremely close mother-child relationship, an unromantic, conflictual marriage characterized by little verbal communication and a peripheral, distant father, may function very differently in other cultures. While we believe that both theories will be greatly enriched by their integration, we caution against the application of either theory outside the cultures in which they were developed.

  14. The substance of cultural evolution: culturally framed systems of social organization.

    PubMed

    Read, Dwight W

    2014-06-01

    Models of cultural evolution need to address not only the organizational aspects of human societies, but also the complexity and structure of cultural idea systems that frame their systems of organization. These cultural idea systems determine a framework within which behaviors take place and provide mutually understood meanings for behavior from the perspective of both agent and recipient that are critical for the coherence of human systems of social organization.

  15. Antifungal activity of Lactobacillus paracasei ssp. tolerans isolated from a sourdough bread culture.

    PubMed

    Hassan, Yousef I; Bullerman, Lloyd B

    2008-01-15

    Lactic acid bacteria were isolated from four different sourdough bread cultures previously investigated for antifungal activity. A total of 116 isolates were obtained and screened for antifungal activity against a battery of molds. The most inhibitory isolate obtained was identified by API 50 CHL and 16s ribosomal RNA genotyping and found to be Lactobacillus paracasei ssp. tolerans. This isolate completely inhibited the growth of Fusarium proliferatum M 5689, M 5991 and Fusarium graminearum R 4053 compared to controls in a dual agar plate assay.

  16. 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

  17. 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…

  18. Evaluation of the OSIRIS video reader as an automated measurement system for the agar disk diffusion technique.

    PubMed

    Kolbert, M; Chegrani, F; Shah, P M

    2004-05-01

    Measurement of inhibition zones by the automated OSIRIS system was compared with manual measurement. In total, 14 176 measurements were made with 352 staphylococcal and 80 Enterobacteriaceae isolates, involving four panels of antibiotics on round and square Mueller-Hinton agar plates, according to the German DIN 58940 recommendations. Variations of +/- 3 mm in zone size measurements were defined as tolerable. Very major errors (i.e., classification of a resistant isolate as susceptible by the OSIRIS system) occurred in < 1% of tests. With staphylococci, the best concordance was recorded for rifampicin (91.3%), moxifloxacin (88.1%), and gentamicin (86.3%), while the concordance on square plates for vancomycin, pristinamycin and kanamycin was 97.2%, 96.1% and 96.0%, respectively. The poorest concordance was for cefuroxime (43.7%) and novobiocin (47.0%) on round plates, and fosfomycin (36.5%) and chloramphenicol (84.0%) on square plates. With Enterobacteriaceae, 100% concordance was recorded for ampicillin, gentamicin and ciprofloxacin on round agar plates, and for gentamicin, cefoxitin and nalidixic acid on square plates. The poorest results were recorded for nalidixic acid (32.5%) and piperacillin (82.5%) on round plates, and for nitrofurantoin (72.5%) and amoxycillin (82.5%) on square plates. It was concluded that the OSIRIS system was a rapid and reliable system for measuring disk susceptibility test results on round and square agar plates.

  19. 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…

  20. 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

  1. 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

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

    PubMed

    Kim, Jae-Seok; Kang, Go-Eun; Kim, Han-Sung; Kim, Hyun Soo; 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.

  3. Performance of Gram staining on blood cultures flagged negative by an automated blood culture system.

    PubMed

    Peretz, A; Isakovich, N; Pastukh, N; Koifman, A; Glyatman, T; Brodsky, D

    2015-08-01

    Blood is one of the most important specimens sent to a microbiology laboratory for culture. Most blood cultures are incubated for 5-7 days, except in cases where there is a suspicion of infection caused by microorganisms that proliferate slowly, or infections expressed by a small number of bacteria in the bloodstream. Therefore, at the end of incubation, misidentification of positive cultures and false-negative results are a real possibility. The aim of this work was to perform a confirmation by Gram staining of the lack of any microorganisms in blood cultures that were identified as negative by the BACTEC™ FX system at the end of incubation. All bottles defined as negative by the BACTEC FX system were Gram-stained using an automatic device and inoculated on solid growth media. In our work, 15 cultures that were defined as negative by the BACTEC FX system at the end of the incubation were found to contain microorganisms when Gram-stained. The main characteristic of most bacteria and fungi growing in the culture bottles that were defined as negative was slow growth. This finding raises a problematic issue concerning the need to perform Gram staining of all blood cultures, which could overload the routine laboratory work, especially laboratories serving large medical centers and receiving a large number of blood cultures.

  4. 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

  5. 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…

  6. 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

  7. 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

  8. 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.

  9. 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

  10. 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.

  11. 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.

  12. Antibacterial activity of four dentin bonding systems.

    PubMed

    Herrera, M; Carrión, P; Bravo, M; Castillo, A

    2000-08-01

    The antibacterial action of bonding systems Gluma 2000, Syntac, Prisma Universal Bond 3, Scotchbond Multipurpose and Prime-Bond was tested against 32 strains of the caries-producing bacteria Streptococcus spp., Lactobacillus spp., Actinomyces spp., Porphyromonas spp. and Clostridium spp. An agar plate diffusion method was used with chlorhexidine as the positive control. Assays were performed in triplicate for each component (primer and adhesive) of the bonding systems. All the adhesives were found to inhibit bacterial growth but with differences in their spectra of action. The sum action of the Scotchbond Multipurpose components were most inhibitory and Prime-Bond was found to be the least effective system.

  13. 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

  14. 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.

  15. Development of Scalable Culture Systems for Human Embryonic Stem Cells

    PubMed Central

    Azarin, Samira M.; Palecek, Sean P.

    2009-01-01

    The use of human pluripotent stem cells, including embryonic and induced pluripotent stem cells, in therapeutic applications will require the development of robust, scalable culture technologies for undifferentiated cells. Advances made in large-scale cultures of other mammalian cells will facilitate expansion of undifferentiated human embryonic stem cells (hESCs), but challenges specific to hESCs will also have to be addressed, including development of defined, humanized culture media and substrates, monitoring spontaneous differentiation and heterogeneity in the cultures, and maintaining karyotypic integrity in the cells. This review will describe our current understanding of environmental factors that regulate hESC self-renewal and efforts to provide these cues in various scalable bioreactor culture systems. PMID:20161686

  16. 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.

  17. Cultural selection drives the evolution of human communication systems.

    PubMed

    Tamariz, Monica; Ellison, T Mark; Barr, Dale J; Fay, Nicolas

    2014-08-01

    Human communication systems evolve culturally, but the evolutionary mechanisms that drive this evolution are not well understood. Against a baseline that communication variants spread in a population following neutral evolutionary dynamics (also known as drift models), we tested the role of two cultural selection models: coordination- and content-biased. We constructed a parametrized mixed probabilistic model of the spread of communicative variants in four 8-person laboratory micro-societies engaged in a simple communication game. We found that selectionist models, working in combination, explain the majority of the empirical data. The best-fitting parameter setting includes an egocentric bias and a content bias, suggesting that participants retained their own previously used communicative variants unless they encountered a superior (content-biased) variant, in which case it was adopted. This novel pattern of results suggests that (i) a theory of the cultural evolution of human communication systems must integrate selectionist models and (ii) human communication systems are functionally adaptive complex systems.

  18. 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

  19. Studies on conjugation of Spirogyra using monoclonal culture.

    PubMed

    Ikegaya, Hisato; Nakase, Takuto; Iwata, Kazuyoshi; Tsuchida, Hideaki; Sonobe, Seiji; Shimmen, Teruo

    2012-05-01

    We succeeded in inducing conjugation of Spirogyra castanacea by incubating algal filaments on agar plate. Conjugation could be induced using clone culture. The scalariform conjugation was generally observed, while lateral conjugation was rarely. When two filaments formed scalariform conjugation, all cells of one filament behaved as male and those of other filament did as female. Very rarely, however, zygospores were formed in both of pair filaments. The surface of conjugation tube was stained with fluorescently labeled-lectins, such as Bandeiraea (Griffonia) simplicifolia lectin (BSL-I) and jacalin. BSL-I strongly stained the conjugation tubes, while weakly did the cell surface of female gamete first and then that of male gamete. Jacalin stained mainly the conjugation tubes. Addition of jacalin inhibited the formation of papilla, suggesting some important role of jacalin-binding material at the initial step of formation of the conjugation tubes.

  20. Method and system of culturing an algal mat

    DOEpatents

    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.

  1. 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.

  2. 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.

  3. 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

  4. Culturally competent systems of care with Latino children and families.

    PubMed

    Dettlaff, Alan J; Rycraft, Joan R

    2009-01-01

    The Latino population represents the fastest growing ethnic minority in the United States. As a result, child welfare agencies need to be prepared to provide culturally appropriate services to Latino families. This paper describes an evaluation of a federally funded initiative designed to train child welfare practitioners in using an existing evidence-based framework--systems of care--with Latino children and families to address the need for culturally competent, community-based services with this population. Results indicate that trained participants responded positively to the systems of care framework, increased their knowledge of systems of care, and reported positive benefits to their clients through using this framework. However, challenges to implementing systems of care were identified. Implications of these findings and the use of systems of care in child welfare are presented.

  5. Developing defined culture systems for human pluripotent stem cells

    PubMed Central

    Valamehr, Bahram; Tsutsui, Hideaki; Ho, Chih-Ming; Wu, Hong

    2013-01-01

    Human pluripotent stem cells hold promising potential in many therapeutics applications including regenerative medicine and drug discovery. Over the past three decades, embryonic stem cell research has illustrated that embryonic stem cells possess two important and distinct properties: the ability to continuously self-renew and the ability to differentiate into all specialized cell types. In this article, we will discuss the continuing evolution of human pluripotent stem cell culture by examining requirements needed for the maintenance of self-renewal in vitro. We will also elaborate on the future direction of the field toward generating a robust and completely defined culture system, which has brought forth collaborations amongst biologists and engineers. As human pluripotent stem cell research progresses towards identifying solutions for debilitating diseases, it will be critical to establish a defined, reproducible and scalable culture system to meet the requirements of these clinical applications. PMID:21916597

  6. 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.

  7. 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

  8. 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

  9. 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'…

  10. 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…

  11. 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…

  12. Pedagogical System of Future Teachers' Professional Thinking Culture Formation

    ERIC Educational Resources Information Center

    Abildina, Saltanat K.; Sarsekeyeva, Zhanar Y.; Aidarbekova, Kulzhan A.; Asetova, Zhannur B.; Adanov, Kuanysbek B.

    2016-01-01

    Research objective is to theoretically justify and to develop a pedagogical system of development of future teachers' professional thinking culture. In the research there are used a set of theoretical methods: systematic analysis of the philosophical, psychological and pedagogical literature on the researched topic; compilation and classification…

  13. 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…

  14. 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.

  15. Slice Culture Modeling of Central Nervous System (CNS) Viral Infection

    PubMed Central

    Dionne, Kalen R.; Tyler, Kenneth L.

    2016-01-01

    The complexity of the central nervous system (CNS) is not recapitulated in cell culture models. Thin slicing and subsequent culture of CNS tissue has become a valued means to study neuronal and glial biology within the context of the physiologically relevant tissue milieu. Modern membrane-interface slice culturing methodology allows straightforward access to both CNS tissue and feeding medium, enabling experimental manipulations and analyses that would otherwise be impossible in vivo. CNS slices can be successfully maintained in culture for up to several weeks for investigation of evolving pathology and long-term intervention in models of chronic neurologic disease. Herein, membrane-interface slice culture models for studying viral encephalitis and myelitis are detailed, with emphasis on the use of these models for investigation of pathogenesis and evaluation of novel treatment strategies. We describe techniques to (1) generate brain and spinal cord slices from rodent donors, (2) virally infect slices, (3) monitor viral replication, (4) assess virally induced injury/apoptosis, (5) characterize “CNS-specific” cytokine production, and (6) treat slices with cytokines/pharmaceuticals. Although our focus is on CNS viral infection, we anticipate that the described methods can be adapted to address a wide range of investigations within the fields of neuropathology, neuroimmunology, and neuropharmacology. PMID:23975824

  16. 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.

  17. 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.

  18. 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

  19. Cellulase production by a solid state culture system

    SciTech Connect

    Kim, J.H.; Hosobuchi, M.; Kishimoto, M.; Seki, T.; Yoshida, T.; Taguchi, H.; Ryu, D.D.Y.

    1985-10-01

    Production of cellulase using solid culture systems of Trichoderma reesei QM9414 and Sporotrichum cellulophilum on wheat bran was studied. By using moisture-controlled solid culture equipment, the effect of water content of wheat bran on cell growth and cellulase production was investigated. Cellular biomass grown on solid substrate was estimated by measuring oxygen consumption rate and glucosamine content the cells. These parameters were shown to have a good linear correlation with the specific growth rate. This reliable method of estimating the cell growth rate enabled us to simulate the enzyme production in a solid culture system by means of multiple linear regression analysis which takes into account of the water content, cell mass, and the oxygen consumption rate as variables. The cell growth and cellulase production were maximized at different water content of the medium. A high water content, 57% for T. reesei and 70% for S. cellulophilum, favored mycelial growth, while the maximum cellulase activity was obtained at a lower water content such as 50% for both fungi. It was observed that cellulase production by T. reesei depended on the culture conditions that support the optimal growth rate for the maximum enzyme production. 14 references.

  20. 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.

  1. Microfluidic cell culture system with on-chip hypoxic conditioning.

    PubMed

    Takano, Atsushi; Tanaka, Masato; Futai, Nobuyuki

    2013-01-01

    We have demonstrated a portable microfluidic cell culture system with multi-gas (CO2 and O2) incubation which we can cultivate under hypoxia without bulky peripheral apparatus such as gas tanks, regulators, and flow controllers. The system contains a chip of 26 mm × 48 mm which is capable to diffuse CO2 and absorb O2 through a gas-permeable wall of nested media reservoir. The media was water-jacketed with aqueous solution containing 0.8 M sodium bicarbonate as CO2 supply and 1 M sodium ascorbate as oxygen scavenger. The partial CO2 pressure (pCO2) in media reservoir stabilized at least 10.2% ± 0.11% for at least 72 hours. The partial O2 pressure (pO2) in the media reservoir decreased to 4.2%. Portable on-chip hypoxic culture of SV40-T2 cells for 72 h was also demonstrated. PMID:24110727

  2. 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

  3. Multiplying steady-state culture in multi-reactor system.

    PubMed

    Erm, Sten; Adamberg, Kaarel; Vilu, Raivo

    2014-11-01

    Cultivation of microorganisms in batch experiments is fast and economical but the conditions therein change constantly, rendering quantitative data interpretation difficult. By using chemostat with controlled environmental conditions the physiological state of microorganisms is fixed; however, the unavoidable stabilization phase makes continuous methods resource consuming. Material can be spared by using micro scale devices, which however have limited analysis and process control capabilities. Described herein are a method and a system combining the high throughput of batch with the controlled environment of continuous cultivations. Microorganisms were prepared in one bioreactor followed by culture distribution into a network of bioreactors and continuation of independent steady state experiments therein. Accelerostat cultivation with statistical analysis of growth parameters demonstrated non-compromised physiological state following distribution, thus the method effectively multiplied steady state culture of microorganisms. The theoretical efficiency of the system was evaluated in inhibitory compound analysis using repeated chemostat to chemostat transfers.

  4. 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

  5. AUTORADIOGRAPHIC ANALYSIS ON AGAR PLATES OF ANTIGENS FROM SUB CELLULAR FRACTIONS OF RAT LIVER SLICES

    PubMed Central

    Morgan, W. S.; Perlmann, P.; Hultin, T.

    1961-01-01

    Slices of rat livers were incubated with 14C amino acids, homogenized, and subjected to differential centrifugation. The microsomes were further extracted with the non-ionic detergent Lubrol W and with EDTA. These extracts and the microsome free "cell sap," freed from the pH 5 precipitable fraction, were subsequently reacted with antisera using agar diffusion techniques. The antisera employed were obtained from rabbits injected with different subcellular fractions of rat liver or with rat serum proteins. When the agar diffusion plates were autoradiographed it was found that some of the precipitates were radioactive while others were not. Control experiments indicated that this labeling was due to the specific incorporation of 14C amino acids into various rat liver antigens during incubation of the slices rather than to a non-specific adsorption of radioactive material to the immunological precipitates. When the slices were incubated with the isotope for up to 30 minutes, the serum proteins which could be extracted from the microsomes with the detergent were strongly labeled, as were a number of additional microsomal antigens of unknown significance. In contrast, the serum proteins present in the cell sap were only weakly labeled. Most of the typical cell sap proteins, both those precipitable and those soluble at pH 5, seemed to remain unlabeled. No consistently reproducible results were obtained with the EDTA extracts of the ribosomal residues remaining after extraction of the microsomes with the detergent. Incubation of the liver slices for longer periods (up to 120 minutes) led to a strong labeling of the serum proteins in the cell sap as well as to the appearance of labeling in additional cell sap proteins. The results are discussed with regard to the subcellular site of synthesis and the metabolism of the different antigens. PMID:13772607

  6. 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.

  7. Use of modified blood agar plate for identification of pathogenic campylobacter species at Mymensingh Medical College .

    PubMed

    Sarkar, S R; Hossain, M A; Pual, S K; Mahmud, M C; Ray, N C; Haque, N

    2014-10-01

    This cross sectional study was carried out from July 2011 to June 2012 in the Department of Microbiology, Mymensingh Medical College to diagnose etiology of diarrhea caused by Campylobacter species. A total of 200 clinically diagnosed diarrheal pediatric patients were included in this study. Among the 200 stool specimens evaluated, 23(11.5%) samples were positive for Campylobacter species, isolation rate was 15(65.2%) in upto 1 year age group and 08(34.7%) in more than 1 year age group. Among 23 positive cases, 20(86.95%) were C. jejuni and 03(13.05%) were C. coli. The prevalence of Campylobacter infection found in the present study was higher below 1 year age group and was very much close to other countries of this Sub continent.

  8. 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.

  9. 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.

  10. 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.

  11. 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

  12. 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

  13. 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);…

  14. 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

  15. 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

  16. 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…

  17. Fine bubble mixing (FBM) culture of E. coli: a highly cost-effective middle scale-size culture system.

    PubMed

    Yasumitsu, Hidetaro; Tajima, Hitoshi; Isobe, Masaharu; Kutsuna, Sinsuke; Kawsar, Sarkar M A; Fujii, Yuki; Kanaly, Robert A; Ozeki, Yasuhiro; Yokota, Eriko

    2013-02-01

    A highly cost-effective and easy-to-assemble cultivation system suitable for middle scale-size culturing of bacterial cells is described. In the culture, from a flat-shaped air-stone with large surface area, fine bubbles are generated with a low-cost air pump available in an aquarium fish shop, and cell-agitation and oxygen supply are efficiently conducted by fine bubbles simultaneously. Growth properties of the cells and their saturation density are comparable to those in a conventional culture system. The expression of recombinant protein was revealed to be similar to conventional methods. The system does not require any expensive machines or equipments. In addition, all equipments except plastic flat-shaped airstone are reusable after sterilization. Due to the low cost, the ease to use and multiple cultivations at once, our system may enable to find better culture conditions, to scale-up with ease and to perform timesaving efficient protein production. PMID:22894161

  18. 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…

  19. 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.

  20. 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

  1. 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

  2. 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.

  3. 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.

  4. 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

  5. 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…

  6. 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

  7. 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.

  8. 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.

  9. 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

  10. 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

  11. Religious Dimensions of World Cultures as Related to the Holt Databank System.

    ERIC Educational Resources Information Center

    Calderwood, Beatrice

    This resource unit, part of a series of guides by the Public Religion Studies Center at Wright State University, correlates with the Holt Databank System "Inquiring About Cultures." The unit covers many facets of many cultures ranging from primitive tribes to complex, urban societies and makes evident the religious dimensions of culture in both…

  12. 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

  13. Fully-automated roller bottle handling system for large scale culture of mammalian cells.

    PubMed

    Kunitake, R; Suzuki, A; Ichihashi, H; Matsuda, S; Hirai, O; Morimoto, K

    1997-01-20

    A fully automatic and continuous cell culture system based on roller bottles is described in this paper. The system includes a culture rack storage station for storing a large number of roller bottles filled with culture medium and inoculated with mammalian cells, mass-handling facility for extracting completed cultures from the roller bottles, and replacing the culture medium. The various component units of the system were controlled either by a general-purpose programmable logic controller or a dedicated controller. The system provided four subsequent operation modes: cell inoculation, medium change, harvesting, and medium change. The operator could easily select and change the appropriate mode from outside of the aseptic area. The development of the system made large-scale production of mammalian cells, and manufacturing and stabilization of high quality products such as erythropoietin possible under total aseptic control, and opened up the door for industrial production of physiologically active substances as pharmaceutical drugs by mammalian cell culture.

  14. Technique for Handling Cover Slips Used in Cell Culture Systems *

    PubMed Central

    Bowne, J. G.; Jochim, Michael M.

    1964-01-01

    A stainless steel, grooved rack was designed to handle large numbers of coverslips used in cell cultures. The rack separated and held the coverslips through the washing process and subsequent manipulations. Slips washed by this method allowed consistently good cell sheets to grow with no signs of cytotoxicity to the cultures. ImagesFIGURE 1.FIGURE 2. PMID:17649500

  15. Forgotten Millions: A Case of Cultural and Systemic Bias

    ERIC Educational Resources Information Center

    Alur, Mithu

    2007-01-01

    In this article the author discusses concepts of exclusion and inclusion and how important it is to understand exclusionary practices in the context and culture specificity of each region to attempt to remove barriers to inclusive education. She specifically discusses the low status given to girls and women in the Indian culture and how this…

  16. 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

  17. 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

  18. 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

  19. Does the concept of safety culture help or hinder systems thinking in safety?

    PubMed

    Reiman, Teemu; Rollenhagen, Carl

    2014-07-01

    The concept of safety culture has become established in safety management applications in all major safety-critical domains. The idea that safety culture somehow represents a "systemic view" on safety is seldom explicitly spoken out, but nevertheless seem to linger behind many safety culture discourses. However, in this paper we argue that the "new" contribution to safety management from safety culture never really became integrated with classical engineering principles and concepts. This integration would have been necessary for the development of a more genuine systems-oriented view on safety; e.g. a conception of safety in which human, technological, organisational and cultural factors are understood as mutually interacting elements. Without of this integration, researchers and the users of the various tools and methods associated with safety culture have sometimes fostered a belief that "safety culture" in fact represents such a systemic view about safety. This belief is, however, not backed up by theoretical or empirical evidence. It is true that safety culture, at least in some sense, represents a holistic term-a totality of factors that include human, organisational and technological aspects. However, the departure for such safety culture models is still human and organisational factors rather than technology (or safety) itself. The aim of this paper is to critically review the various uses of the concept of safety culture as representing a systemic view on safety. The article will take a look at the concepts of culture and safety culture based on previous studies, and outlines in more detail the theoretical challenges in safety culture as a systems concept. The paper also presents recommendations on how to make safety culture more systemic. PMID:24275532

  20. Does the concept of safety culture help or hinder systems thinking in safety?

    PubMed

    Reiman, Teemu; Rollenhagen, Carl

    2014-07-01

    The concept of safety culture has become established in safety management applications in all major safety-critical domains. The idea that safety culture somehow represents a "systemic view" on safety is seldom explicitly spoken out, but nevertheless seem to linger behind many safety culture discourses. However, in this paper we argue that the "new" contribution to safety management from safety culture never really became integrated with classical engineering principles and concepts. This integration would have been necessary for the development of a more genuine systems-oriented view on safety; e.g. a conception of safety in which human, technological, organisational and cultural factors are understood as mutually interacting elements. Without of this integration, researchers and the users of the various tools and methods associated with safety culture have sometimes fostered a belief that "safety culture" in fact represents such a systemic view about safety. This belief is, however, not backed up by theoretical or empirical evidence. It is true that safety culture, at least in some sense, represents a holistic term-a totality of factors that include human, organisational and technological aspects. However, the departure for such safety culture models is still human and organisational factors rather than technology (or safety) itself. The aim of this paper is to critically review the various uses of the concept of safety culture as representing a systemic view on safety. The article will take a look at the concepts of culture and safety culture based on previous studies, and outlines in more detail the theoretical challenges in safety culture as a systems concept. The paper also presents recommendations on how to make safety culture more systemic.

  1. 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)).

  2. 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

  3. A feasible enzyme-linked immunosorbent assay system using monoclonal and polyclonal antibodies against glucosyltransferase-B from Streptococcus mutans.

    PubMed

    Shinozaki-Kuwahara, Noriko; Hashizume-Takizawa, Tomomi; Hirasawa, Masatomo; Takada, Kazuko

    2012-06-01

    Streptococcus mutans has been considered the principal etiological agent of dental caries in humans. S. mutans can secrete three kinds of glucosyltransferases (GTFs). One of these, GTF-B, which synthesizes water-insoluble glucans from sucrose, has been considered to be one of the most important factors of cariogenic dental plaque formation. Therefore, determination of whether GTF-B is present in plaque and saliva samples may contribute to the evaluation of individual virulence potential (caries risk). The aim of this study was to develop a feasible enzyme-linked immunosorbent assay (ELISA) for the routine quantification of GTF-B in plaque-derived cultures and clinical samples, and to apply this assay to an epidemiological study. To determine the presence of GTF-B in plaque samples, a sandwich-ELISA was devised, consisting of mouse monoclonal and rabbit polyclonal antibodies against GTF-B and a horseradish peroxidase-conjugated anti-rabbit antibody. The developed ELISA allowed for quantification of the amounts of purified GTF-B with satisfactory sensitivity and specificity; this method was not affected by other components such as plaque and saliva. Plaque samples from healthy volunteers were examined using this ELISA method and microbial analysis to apply the assay to an epidemiological study. A correlation was observed between the amount of extracted GTF-B and S. mutans levels as determined by ELISA and cultivated with Mitis Salivarius Bacitracin agar plates derived from plaque samples, although there were some exceptions. In this regard, this ELISA system has the advantage of estimating both the individual numbers of S. mutans and the productivity of GTF-B, namely, the cariogenic potential of S. mutans simultaneously. These results indicate that this ELISA method is a useful tool for the diagnosis of caries risk.

  4. Ethanol production from starch by a coimmobilized mixed culture system of Aspergillus awamori and Zymomonas mobilis

    SciTech Connect

    Tanaka, H.; Kurosawa, H.; Murakami, H.

    1986-01-01

    The production of ethanol from starch by a coimmobilized mixed culture system of aerobic and anaerobic microorganisms in Ca-alginate gel beads was investigated. The mold Aspergillus awamori was used as an aerobic amylolytic microorganism and an anaerobic bacterium, Zymomonas mobilis, as an ethanol producer. By controlling the mixing ratio of the microorganisms in the inoculum size, a desirable coimmobilized mixed culture system, in which the aerobic mycelia grew on and near the oxygen-rich surface of the gel beads while the anaerobic bacterial cells mainly grew in the oxygen-deficient central part of the gel beads, was naturally established under the aerobic culture conditions, and ethanol could be directly produced from starch by the system. The ethanol productivity by the system in flask culture was particularly affected by the shear stress (dependent on the shaking speed) which controlled the mycelial growth on the surface of the gel beads. Under optimum culture conditions in the flask culture, the glucose produced was instantly consumed, and was not observed in the culture broth; the final concentration of ethanol produced from 100 g/L starch was 25 g/L and the yield coefficient for ethanol, Y/sub p/s/, was 0.38. The ethanol productivity by the coimmobilized mixed culture system was compared with those by other various culture systems and the advantages of the system were clarified.

  5. A microdroplet cell culture based high frequency somatic embryogenesis system for pigeonpea, Cajanus cajan (L.) Millsp.

    PubMed

    Kumar, Nagan Udhaya; Gnanaraj, Muniraj; Sindhujaa, Vajravel; Viji, Maluventhen; Manoharan, Kumariah

    2015-09-01

    A protocol for high frequency production of somatic embryos was worked out in pigeonpea, Cajanus cajan (L.) Millsp. The protocol involved sequential employment of embryogenic callus cultures, low density cell suspension cultures and a novel microdroplet cell culture system. The microdroplet cell cultures involved culture of a single cell in 10 μI of Murashige and Skoog's medium supplemented with phytohormones, growth factors and phospholipid precursors. By employing the microdroplet cell cultures, single cells in isolation were grown into cell clones which developed somatic embryos. Further, 2,4-dichlorophenoxyacetic acid, kinetin, polyethylene glycol, putrescine, spermine, spermidine, choline chloride, ethanolamine and LiCl were supplemented to the low density cell suspension cultures and microdroplet cell cultures to screen for their cell division and somatic embryogenesis activity. Incubation of callus or the inoculum employed for low density cell suspension cultures and microdroplet cell cultures with polyethylene glycol was found critical for induction of somatic embryogenesis. Somatic embryogenesis at a frequency of 1.19, 3.16 and 6.51 per 10(6) cells was achieved in the callus, low density cell suspension cultures and microdroplet cell cultures, respectively. Advantages of employing microdroplet cell cultures for high frequency production of somatic embryos and its application in genetic transformation protocols are discussed. PMID:26548080

  6. A microdroplet cell culture based high frequency somatic embryogenesis system for pigeonpea, Cajanus cajan (L.) Millsp.

    PubMed

    Kumar, Nagan Udhaya; Gnanaraj, Muniraj; Sindhujaa, Vajravel; Viji, Maluventhen; Manoharan, Kumariah

    2015-09-01

    A protocol for high frequency production of somatic embryos was worked out in pigeonpea, Cajanus cajan (L.) Millsp. The protocol involved sequential employment of embryogenic callus cultures, low density cell suspension cultures and a novel microdroplet cell culture system. The microdroplet cell cultures involved culture of a single cell in 10 μI of Murashige and Skoog's medium supplemented with phytohormones, growth factors and phospholipid precursors. By employing the microdroplet cell cultures, single cells in isolation were grown into cell clones which developed somatic embryos. Further, 2,4-dichlorophenoxyacetic acid, kinetin, polyethylene glycol, putrescine, spermine, spermidine, choline chloride, ethanolamine and LiCl were supplemented to the low density cell suspension cultures and microdroplet cell cultures to screen for their cell division and somatic embryogenesis activity. Incubation of callus or the inoculum employed for low density cell suspension cultures and microdroplet cell cultures with polyethylene glycol was found critical for induction of somatic embryogenesis. Somatic embryogenesis at a frequency of 1.19, 3.16 and 6.51 per 10(6) cells was achieved in the callus, low density cell suspension cultures and microdroplet cell cultures, respectively. Advantages of employing microdroplet cell cultures for high frequency production of somatic embryos and its application in genetic transformation protocols are discussed.

  7. 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.

  8. Direct identification of bacterial isolates in blood cultures by using a DNA probe.

    PubMed

    Davis, T E; Fuller, D D

    1991-10-01

    This study involved the rapid, direct identification of Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, Haemophilus influenzae, Enterococcus sp., and Streptococcus agalactiae from positive blood culture bottles (BACTEC, Johnston Laboratories, Inc.) by using the AccuProbe (Gen-Probe, San Diego, Calif.) culture confirmation test. This method uses a chemiluminescent DNA probe that detects the rRNA of the target organisms. The manufacturer's instructions were modified to use a pellet of bacteria made directly from positive blood culture broth rather than a colony from an agar plate. Two separate procedures of selective centrifugation were employed in order to obtain the pellet. The first utilized a routine clinical centrifuge and a large volume of broth (10 to 12 ml) from the blood culture bottle. The second method used a microcentrifuge and less volume (1 to 1.5 ml). A total of 196 clinical specimens taken directly from positive blood culture broths were correctly identified by AccuProbe from pellets made by using the clinical centrifuge technique, while 166 clinical specimens used as negative controls failed to show hybridization. The microcentrifuge technique for obtaining pellets was performed on 105 patient specimens, and all were correctly identified. When combined with the microcentrifuge technique for pellet preparation, the AccuProbe test has several advantages: (i) direct identification of bacteria from blood culture broths, (ii) rapid turn-around time (30 min), (iii) simplicity of the procedure, and (iv) relative low cost.

  9. 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…

  10. 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.

  11. Growth detection failures by the nonradiometric Bactec MGIT 960 mycobacterial culture system.

    PubMed

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

    2012-06-01

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

  12. 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.

  13. A dual-color luciferase assay system reveals circadian resetting of cultured fibroblasts by co-cultured adrenal glands.

    PubMed

    Noguchi, Takako; Ikeda, Masaaki; Ohmiya, Yoshihiro; Nakajima, Yoshihiro

    2012-01-01

    In mammals, circadian rhythms of various organs and tissues are synchronized by pacemaker neurons in the suprachiasmatic nucleus (SCN) of the hypothalamus. Glucocorticoids released from the adrenal glands can synchronize circadian rhythms in other tissues. Many hormones show circadian rhythms in their plasma concentrations; however, whether organs outside the SCN can serve as master synchronizers to entrain circadian rhythms in target tissues is not well understood. To further delineate the function of the adrenal glands and the interactions of circadian rhythms in putative master synchronizing organs and their target tissues, here we report a simple co-culture system using a dual-color luciferase assay to monitor circadian rhythms separately in various explanted tissues and fibroblasts. In this system, circadian rhythms of organs and target cells were simultaneously tracked by the green-emitting beetle luciferase from Pyrearinus termitilluminans (ELuc) and the red-emitting beetle luciferase from Phrixothrix hirtus (SLR), respectively. We obtained tissues from the adrenal glands, thyroid glands, and lungs of transgenic mice that expressed ELuc under control of the promoter from a canonical clock gene, mBmal1. The tissues were co-cultured with Rat-1 fibroblasts as representative target cells expressing SLR under control of the mBmal1 promoter. Amplitudes of the circadian rhythms of Rat-1 fibroblasts were potentiated when the fibroblasts were co-cultured with adrenal gland tissue, but not when co-cultured with thyroid gland or lung tissue. The phases of Rat-1 fibroblasts were reset by application of adrenal gland tissue, whereas the phases of adrenal gland tissue were not influenced by Rat-1 fibroblasts. Furthermore, the effect of the adrenal gland tissue on the fibroblasts was blocked by application of a glucocorticoid receptor (GR) antagonist. These results demonstrate that glucocorticoids are strong circadian synchronizers for fibroblasts and that this co-culture

  14. [Evaluation for anaerobic culture system: Anoxomat Mart II].

    PubMed

    Kikuchi, Yuji; Sasaki, Hiromi; Furuhata, Yukie; Tazawa, Yoko; Horiuchi, Hajime; Okada, Jun

    2007-01-01

    Anoxomat Mart II (Mart Microbiology BV, Lichtenvooorde, Netherlands, Central Scientific Commerce Inc., Tokyo, Japan) is an anaerobic jar apparatus which uses a vacuum pump in combination with catalyst as gas replacement procedure to remove all traces of oxygen. As we had a chance to use Anoxomat Mart II, we compared it with other two anaerobic culture methods; namely AnaeroPack anaero (Mitsubishi Gas Chemical Co., Tokyo, Japan) which employs anaerobic jar method, and Concept400 (RUSKINN TECHNOLOGY LTD, England; Central Scientific Commerce INc., Tokyo, Japan) which uses anaerobic chamber method. We used 10 different species of anaerobic bacteria obtained from ATCC. One strain each of 10 species was cultured and examined for measurement of the sensitibity of an anaerobic indicator, th number of bacteria after 48 hour culture, the diameter of colonies, and MIC value. As a result, the time to reach the anaerobic condition was around 30 minutes by the Mart II against around 60 minutes by the AnaeroPack anaero. There was no difference concerning the number of bacteria after 48 hour culture among three methods. But anaerobic bacteria cultured by Mart II tended to make bigger colonies compared to other two methods in the 5 strains out of 9, except for one strain in which the diameter of colonies could not be measured. On the other hand, the comparison of MIC value showed good correlation in 11 antibiotics out of 12 among three methods. The MIC value of 11 antibiotics fitted within 1-fold difference, and 2-fold difference was observed in only one antibiotic. Mart II is so small that it does cheep consumables. From these reasons, we concluded that Mart II can be one of the useful anerobic culture methods.

  15. FIZICS: fluorescent imaging zone identification system, a novel macro imaging system.

    PubMed

    Skwish, Stephen; Asensio, Francisco; King, Greg; Clarke, Glenn; Kath, Gary; Salvatore, Michael J; Dufresne, Claude

    2004-12-01

    Constantly improving biological assay development continues to drive technological requirements. Recently, a specification was defined for capturing white light and fluorescent images of agar plates ranging in size from the NUNC Omni tray (96-well footprint, 128 x 85 mm) to the NUNC Bio Assay Dish (245 x 245 mm). An evaluation of commercially available products failed to identify any system capable of fluorescent macroimaging with discrete wavelength selection. To address the lack of a commercially available system, a custom imaging system was designed and constructed. This system provides the same capabilities of many commercially available systems with the added ability to fluorescently image up to a 245 x 245 mm area using wavelengths in the visible light spectrum.

  16. 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

  17. 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

  18. In vivo bone regeneration using tubular perfusion system bioreactor cultured nanofibrous scaffolds.

    PubMed

    Yeatts, Andrew B; Both, Sanne K; Yang, Wanxun; Alghamdi, Hamdan S; Yang, Fang; Fisher, John P; 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(ε-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 mm(2) at 21 days compared to 0.99 ± 0.43 mm(2) and 0.50 ± 0.29 mm(2) 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 mm(2). Defects implanted with statically cultured and acellular scaffolds had a new bone area of 1.26 ± 0.43 mm(2) and 1.19 ± 0.33 mm(2), 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.

  19. 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.

  20. In Vitro Cultivation of 'Unculturable' Oral Bacteria, Facilitated by Community Culture and Media Supplementation with Siderophores.

    PubMed

    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.

  1. 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

  2. In Vitro Cultivation of 'Unculturable' Oral Bacteria, Facilitated by Community Culture and Media Supplementation with Siderophores.

    PubMed

    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

  3. 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…

  4. 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.

  5. 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…

  6. 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…

  7. Culture & Clay: The Reshaping of an Educational System.

    ERIC Educational Resources Information Center

    Levy, Linda Jean

    The validity of Bilingual Education in a multi-cultural preschool setting is examined and supported. Further, a discussion of how mental health may be linked to the recognition of a child's native language, in his aquisition of a positive self-concept, is delineated. Additionally, theoretical information is supported by a survey completed in 1976,…

  8. 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.

  9. 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.

  10. 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

  11. 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.

  12. Effective enrichment of cholangiocarcinoma secretomes using the hollow fiber bioreactor culture system.

    PubMed

    Weeraphan, Churat; Diskul-Na-Ayudthaya, Penchatr; Chiablaem, Khajeelak; Khongmanee, Amnart; Chokchaichamnankit, Daranee; Subhasitanont, Pantipa; Svasti, Jisnuson; Srisomsap, Chantragan

    2012-09-15

    The Northeastern region of Thailand is well known to have high incidence of bile duct cancer known as cholangiocarcinoma. So there is a continued need to improve diagnosis and treatment, and discovery of biomarkers for early detection of bile duct cancer should greatly improve treatment outcome for these patients. The secretome, a collection of proteins secreted from cells, is a useful source for identifying circulating biomarkers in blood secreted from cancer cells. Here a Hollow Fiber Bioreactor culture system was used for enrichment of cholangiocarcinoma secretomes, since this culture system mimics the dense three-dimensional microenvironment of the tumor found in vivo. Two-dimensional fluorescence difference gel electrophoresis using a sensitive Fluor saturation dye staining, followed by LC/MS/MS, was used to compare protein expression in the secretomes of cells cultured in the Hollow Fiber system and cells cultured in the monolayer culture system. For the first time, the 2D-patterns of cholangiocarcinoma secretomes from the two culture systems could be compared. The Hollow Fiber system improved the quality and quantity of cholangiocarcinoma secreted proteins compared to conventional monolayer system, showing less interference by cytoplasmic proteins and yielding more secreted proteins. Overall, 75 spots were analyzed by LC/MS/MS and 106 secreted proteins were identified. Two novel secreted proteins (C19orf10 and cystatin B) were found only in the Hollow Fiber system and were absent from the traditional monolayer culture system. Among the highly expressed proteins, 22 secreted soluble proteins were enriched by 5 fold in Hollow Fiber system compared to monolayer culture system. The Hollow Fiber system is therefore useful for preparing a wide range of proteins from low-abundance cell secretomes.

  13. 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.

  14. 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…

  15. 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…

  16. 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…

  17. Experience of changing between signal and Bactec 9240 blood culture systems in a children's hospital.

    PubMed Central

    Gray, J; Brockwell, M; Das, I

    1998-01-01

    AIM: To compare experience of positive blood cultures in successive years before and after changing from Signal (Unipath) to Bactec 9240 (Becton Dickinson) blood culture systems. METHODS: Analysis of data collected prospectively on 7967 Signal and 7062 Bactec blood culture sets. RESULTS: Significant growths occurred in 5.7% of Signal and 8.9% of Bactec cultures; 33.0% more significant isolates and 24.0% more episodes of bacteraemia were detected in the second year, following introduction of the Bactec system. Inpatient hospital activity increased by 8.2% between the first and second years, although the numbers of blood cultures received actually fell by 11.4%. There were striking increases in numbers of isolates of coagulase negative staphylococci (47.7%) and Enterobacteriaceae (56.8%) from Bactec cultures. Two anaerobic bacteraemias were detected in Signal blood cultures, whereas none was detected by the Bactec system, despite 12.1% of sets including an anaerobic bottle. Of significant positive cultures, 90.2% were detected within one day with the Bactec 9240, compared with only 50.0% of Signal cultures; 20.7% of significant positive Signal blood cultures were detected only on terminal subculture. Microorganisms that were not significant were isolated from 5.1% Signal and 3.8% Bactec cultures. CONCLUSIONS: Compared with the Signal system, the Bactec 9240 offers markedly more rapid and sensitive detection of bacteraemia, together with a lower rate of non-significant isolates. However, using a single PEDS PLUS/F bottle the few episodes of anaerobic bacteraemia that occur in children are likely to be missed. PMID:9659243

  18. Organoid culture systems for prostate epithelial and cancer tissue.

    PubMed

    Drost, Jarno; Karthaus, Wouter R; Gao, Dong; Driehuis, Else; Sawyers, Charles L; Chen, Yu; Clevers, Hans

    2016-02-01

    This protocol describes a strategy for the generation of 3D prostate organoid cultures from healthy mouse and human prostate cells (either bulk or FACS-sorted single luminal and basal cells), metastatic prostate cancer lesions and circulating tumor cells. Organoids derived from healthy material contain the differentiated luminal and basal cell types, whereas organoids derived from prostate cancer tissue mimic the histology of the tumor. We explain how to establish these cultures in the fully defined serum-free conditioned medium that is required to sustain organoid growth. Starting with the plating of digested tissue material, full-grown organoids can usually be obtained in ∼2 weeks. The culture protocol we describe here is currently the only one that allows the growth of both the luminal and basal prostatic epithelial lineages, as well as the growth of advanced prostate cancers. Organoids established using this protocol can be used to study many different aspects of prostate biology, including homeostasis, tumorigenesis and drug discovery.

  19. 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

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

    PubMed

    Jin, Peng; Wang, Shiow Y; Gao, Haiyan; Chen, Hangjun; Zheng, Yonghua; Wang, Chien Y

    2012-05-01

    The effects of cultural system and essential oil treatment on antioxidant capacities in raspberries were evaluated. Raspberries were hand-harvested from organic and conventional farms in Maryland, USA, and were treated with essential oil including carvacrol, anethole, cinnamic acid, perillaldehyde, cinnamaldehyde, and linalool. Results from this study showed that raspberries grown from organic culture exhibited higher value of antioxidant capacities and individual flavonoids contents. Moreover, the organic culture also enhanced the activities of antioxidant enzymes. In addition, essential oil treatments promoted the antioxidant enzymes activities and antioxidant capacities of raspberries, and the most effective compound was perillaldehyde. In conclusion, raspberries produced from organic culture contained significantly higher antioxidant capacities than those produce from conventional culture. Postharvest essential oil treatments have positive effect on enhancing antioxidant capacities in raspberries from both organic and conventional cultures.

  1. 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

  2. Proliferation and differentiation of mesenchymal stem cells in a three- dimensional culture system

    NASA Astrophysics Data System (ADS)

    Sun, S.; Hu, J.; Long, M.; Tao, Z.

    Mesenchymal stem cells (MSCs) have multi-differentiation potential and retain the capacity to proliferate and differentiate in vitro, which in turn holds the promise of being able to repair or replace damaged cells or tissues. Since MSCs are rare in amount in vivo, abundant cells usually need be obtained in time in clinic. T herefore, proliferation and differentiation of MSCs in vitro are necessary and important for future applications. Most current studies o MSCs are focused on the cellular andn molecular biology using a two-dimension (2-D) static culture system at unit gravity. The gravity-induced 2D culture of MSCs could potentially not reflect cell-cell- contacts important for proliferation and differentiation of MSCs in vivo. Here we developed a method to proliferate MSCs by using the rotating three-dimensional (3- D) culture system, which can provide low shear, 3-D environment with simulated microgravity. MSCs from human bone marrow were prepared on microcarrier beads and then were seeded in the 3-D culture system. Various rotation conditions were tested to screen out the most suitable one for proliferation of MSCs. 2-D cultures were prepared in routine cell culture dishes as a control. All cultures were uniformly inoculated and medium exchanged at standard intervals. Results were compared with microscopic and immunochemistrical techniques. The differentiation capacity of proliferated MSCs were also tested through induced differentiation experiments. It is found that simulated microgravity and three-dimensional culture condition is an active factor for proliferation of MSCs.

  3. Detection and phylogenetic analysis of coastal bioaerosols using culture dependent and independent techniques

    NASA Astrophysics Data System (ADS)

    Urbano, R.; Palenik, B.; Gaston, C. J.; Prather, K. A.

    2010-08-01

    Bioaerosols are emerging as important yet poorly understood players in atmospheric processes. Microorganisms in the atmosphere have great potential to impact chemical and physical processes that influence global climateby participating in both ice nucleation and cloud droplet formation. The role of microorganisms in atmospheric processes is thought to be species-specific and, potentially, dependent on the viability of the cell; however, few simultaneous measurements of both parameters exist. Using a coastal pier monitoring site as a sampling platform to investigate the exchange of airborne microorganisms at the air-sea interface, culture independent (i.e. DNA clone libraries from filters) and culture dependent approaches (i.e. agar plates) were combined with 18S rRNA and 16S rRNA gene targeting to determine the microbial diversity. The results indicate that in these coastal air samples two fungal phyla, Basidiomycota and Ascomycota, predominate among eukaryotes while Firmicutes and Proteobacteria predominate among bacteria. Furthermore, our culture dependent study verifies the viability of microbes from all four phyla detected through our culture independent study. Contrary to our expectations and despite oceanic air mass sources, common marine planktonic bacteria and phytoplankton were not abundantly found in our air samples indicating the potential importance of bioaerosols derived from beaches and/or coastal erosion processes.

  4. Detection and phylogenetic analysis of coastal bioaerosols using culture dependent and independent techniques

    NASA Astrophysics Data System (ADS)

    Urbano, R.; Palenik, B.; Gaston, C. J.; Prather, K. A.

    2011-02-01

    Bioaerosols are emerging as important yet poorly understood players in atmospheric processes. Microorganisms can impact atmospheric chemistry through metabolic reactions and can potentially influence physical processes by participating in ice nucleation and cloud droplet formation. Microbial roles in atmospheric processes are thought to be species-specific and potentially dependent on cell viability. Using a coastal pier monitoring site as a sampling platform, culture-dependent (i.e. agar plates) and culture-independent (i.e. DNA clone libraries from filters) approaches were combined with 18S rRNA and 16S rRNA gene targeting to obtain insight into the local atmospheric microbial composition. From 13 microbial isolates and 42 DNA library clones, a total of 55 sequences were obtained representing four independent sampling events. Sequence analysis revealed that in these coastal samples two fungal phyla, Ascomycota and Basidiomycota, predominate among eukaryotes while Firmicutes and Proteobacteria predominate among bacteria. Furthermore, our culture-dependent study verifies the viability of microbes from all four phyla detected through our culture-independent study. Contrary to our expectations and despite oceanic air mass sources, common marine planktonic bacteria and phytoplankton were not typically found. The abundance of terrestrial and marine sediment-associated microorganisms suggests a potential importance for bioaerosols derived from beaches and/or coastal erosion processes.

  5. 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

  6. 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

  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. In vitro culture of previously uncultured oral bacterial phylotypes.

    PubMed

    Thompson, Hayley; Rybalka, Alexandra; Moazzez, Rebecca; Dewhirst, Floyd E; Wade, William G

    2015-12-01

    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.

  9. In vitro culture of previously uncultured oral bacterial phylotypes.

    PubMed

    Thompson, Hayley; Rybalka, Alexandra; Moazzez, Rebecca; Dewhirst, Floyd E; Wade, William G

    2015-12-01

    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

  10. 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

  11. [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

  12. [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.

  13. Manufacture of biopharmaceutical proteins by mammalian cell culture systems.

    PubMed

    Tolbert, W R

    1990-01-01

    In the last several years, dramatic advances have been in the development of new biopharmaceuticals including monoclonal antibodies for diagnosis and treatment and such genetically engineered proteins as tPA, Factor VIIIc, erythropoietin and soluble CD4, an anti-AIDS protein. Currently, there are several hundred such candidate drugs in human clinical trials. In most cases, these protein-based drugs will require manufacture by mammalian cell culture due to the inability of lower organisms to properly glycosylate, fold, make correct disulfide bonds and secrete active biomolecular forms. The need for large scale production from cell culture will greatly increase as more of the products in clinical trials are approved for commercial production. This will require significant reduction in manufacturing costs per gram, concomitant with increased capacity to hundreds or perhaps even thousands of kilograms annually. As an example, Invitron's multi-reactor manufacturing facility has operated at greater than one-half million liters per year and has experience with more than 250 mammalian cell lines for producing protein drug products.

  14. 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.

  15. Control of galactosylated glycoforms distribution in cell culture system.

    PubMed

    McCracken, Neil A; Kowle, Ronald; Ouyang, Anli

    2014-01-01

    Cell culture process conditions including media components and bioreactor operation conditions have a profound impact on recombinant protein quality attributes. Considerable changes in the distribution of galactosylated glycoforms (G0F, G1F, and G2F) were observed across multiple CHO derived recombinant proteins in development at Eli Lilly and Company when switching to a new chemically defined (CD) media platform condition. In the new CD platform, significantly lower G0F percentages and higher G1F and G2F were observed. These changes were of interest as glycosylation heterogeneity can impact the effectiveness of a protein. A systematic investigation was done to understand the root cause of the change and control strategy for galactosylated glycoforms distribution. It was found that changes in asparagine concentration could result in a corresponding change in G0F, G1F, and G2F distribution. A follow-up study examined a wider range of asparagine concentration and it was found that G0F, G1F, and G2F percentage could be titrated by adjusting asparagine concentration. The observed changes in heterogeneity from changing asparagine concentration are due to resulting changes in ammonium metabolism. Further study ascertained that different integrated ammonium level during the cell culture process could control G0F, G1F, and G2F percentage distribution. A mechanism hypothesis is proposed that integrated ammonium level impacts intracellular pH, which further regulates β-1, 4 galactosyltransferase activity. PMID:24692242

  16. 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

  17. 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.

  18. 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

  19. Development of practical investigation system for cultural properties based on a projector-camera system

    NASA Astrophysics Data System (ADS)

    Miyata, Kimiyoshi

    2009-01-01

    Huge number of historical materials and cultural properties in museums are investigated based on scientific and chemical analysis techniques, however these techniques require specific equipments with difficult operations. In this research, a practical investigation system is developed to provide convenient way for investigating color information in variety of materials as the first step of the investigation process. The system consists of a data projector and a digital camera, and the system is examined to detect metameric color area as an example of investigation purposes. In this research, the data projector has a role of illuminator to yield illuminant metamerism, and the camera takes sequential images under different illumination colors created by the projector. In the experiment, seven colors of illumination are created by the projector, then images are taken under each colored illumination. The Euclid distance in the RGB space between a predetermined reference pixel and test pixels is calculated in each of taken images. These distances are compared with thresholds determined by using a metamerism test chart. The proposed system was examined for the test charts and Japanese authentic wooden prints, and the experimental results showed that the system could offer convenience as the first investigation of the materials.

  20. 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

  1. 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

  2. Short report: Failure of Burkholderia pseudomallei to grow in an automated blood culture system.

    PubMed

    Teerawattanasook, Nittaya; Limmathurotsakul, Direk; Day, Nicholas P J; Wuthiekanun, Vanaporn

    2014-12-01

    We compared the organisms isolated from 30,210 pairs of blood culture bottles by using BacT/Alert system and the conventional system. Overall, 2,575 (8.5%) specimens were culture positive for pathogenic organisms. The sensitivity for detection of pathogenic organisms with the BACT/Alert system (85.6%, 2,203 of 2,575) was significantly higher than that with the conventional method (74.1%, 1,908 of 2,575; P < 0.0001). However, Burkholderia pseudomallei was isolated less often with the BacT/ALERT system (73.5%, 328 of 446) than with the conventional system (90.3%, 403 of 446; P < 0.0001). This finding suggests that use of the conventional culture method in conjunction with the BacT/Alert system may improve the isolation rate for B. pseudomallei in melioidosis-endemic areas.

  3. 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.

  4. Two-dimensional gel electrophoresis for controlling and comparing culture supernatants of mammalian cell culture productions systems.

    PubMed

    Wimmer, K; Harant, H; Reiter, M; Blüml, G; Gaida, T; Katinger, H

    1994-01-01

    A recombinant Chinese hamster ovary cell line, producing human erythropoietin, was cultivated in a continuous mode in a stirred tank reactor applying different dilution rates. In order to monitor the stability of this expression system, product and non-product proteins of the cell culture supernatant were analyzed by two-dimensional electrophoresis. The consistency of the isoforms of the recombinant product was determined by western blot combined with specific staining. The same cell line was propagated in a high cell density cultivation system based on macro-cell-aggregates. The patterns of secreted proteins of the cell line cultivated in the different systems were compared in order to detect modifications in protein expression of the product and of non product proteins relevant for cell culture supernatant. Hardly any alterations in two-dimensional pattern were detectable. The isoforms of erythropoietin, as well as the overall pattern of secreted proteins, detectable with the two-dimensional electrophoresis method were remarkably stable under different cultivation conditions.

  5. Blood culture examinations at a community hospital without a microbiology laboratory: using an automated blood culture system and performing a Gram stain on positive culture bottles in the institution.

    PubMed

    Saito, Takashi; Aoki, Yoji; Mori, Yoshihiro; Kohi, Fumikazu

    2004-08-01

    To elucidate the existence of microorganisms from blood culture bottles in hospitals without a microbiology laboratory, we changed the system of blood culture examinations. The Oxoid signal blood culture system and submission of all blood cultures to the clinical testing industry was used from July 2002 to December 2002 (first period). Use of the BacT/Alert system and performing of Gram stain for positive culture bottles in our institutions was conducted from January 2003 to June 2003 (latter period). A total of 210 and 193 blood cultures were processed during the first and latter periods, respectively. There were 40 (19.0%) positive cultures in the first period and 32 (16.6%) positive cultures in the latter period. The times from the specimen collection to the Gram stain result that were required were 3.8 and 1.0 days in the first period and the latter period, respectively. The times required for the final report of the blood cultures in the first period and in the latter period were 5.8 and 4.9 days, respectively. We conclude that using a continuous monitoring, automated blood culture system and performing Gram stain for positive culture bottles in institutions without microbiology laboratories may be useful for medical doctors to rapidly determine the existence of microorganisms and to begin adequate antiinfective therapy.

  6. Spatially monitoring oxygen level in 3D microfabricated cell culture systems using optical oxygen sensing beads.

    PubMed

    Wang, Lin; Acosta, Miguel A; Leach, Jennie B; Carrier, Rebecca L

    2013-04-21

    Capability of measuring and monitoring local oxygen concentration at the single cell level (tens of microns scale) is often desirable but difficult to achieve in cell culture. In this study, biocompatible oxygen sensing beads were prepared and tested for their potential for real-time monitoring and mapping of local oxygen concentration in 3D micro-patterned cell culture systems. Each oxygen sensing bead is composed of a silica core loaded with both an oxygen sensitive Ru(Ph2phen3)Cl2 dye and oxygen insensitive Nile blue reference dye, and a poly-dimethylsiloxane (PDMS) shell rendering biocompatibility. Human intestinal epithelial Caco-2 cells were cultivated on a series of PDMS and type I collagen based substrates patterned with micro-well arrays for 3 or 7 days, and then brought into contact with oxygen sensing beads. Using an image analysis algorithm to convert florescence intensity of beads to partial oxygen pressure in the culture system, tens of microns-size oxygen sensing beads enabled the spatial measurement of local oxygen concentration in the microfabricated system. Results generally indicated lower oxygen level inside wells than on top of wells, and local oxygen level dependence on structural features of cell culture surfaces. Interestingly, chemical composition of cell culture substrates also appeared to affect oxygen level, with type-I collagen based cell culture systems having lower oxygen concentration compared to PDMS based cell culture systems. In general, results suggest that oxygen sensing beads can be utilized to achieve real-time and local monitoring of micro-environment oxygen level in 3D microfabricated cell culture systems.

  7. 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

  8. 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…

  9. 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.

  10. 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

  11. Population cell differentiation of Serratia marcescens on agar surface and in broth culture.

    PubMed

    Lai, H C; Lai, M J; Lin-Chao, S; Lu, K T; Ho, S W

    1997-11-01

    The bacterium Serratia marcescens shows population surface migration (swarming) phenomenum on an LB swarming plate, and differentiated cells can be observed at the swarming front. How the cell population differentiates during swarming on the agar surface is not known, neither is it clear whether cells with differentiated characteristics can be observed in broth culture. To monitor the population cell differentiation in a highly sensitive way without cell destruction, experiments were designed using bacterial luciferase genes luxAB as the reporter genes to allow direct monitoring of the differentiating cells through bioluminescence. An isogenic S. marcescens strain was constructed with luxAB under the control of the promoter of flagellin gene hag (phag::luxAB). Patterns of cell differentiation were monitored either by direct X-ray film exposure and/or by Autolumat luminometer detection. Results show that population cell differentiation on the agar surface occurs first in a temporal and then spatial way during colonial growth. It was also found that cells harvested from both the spreading agar plate and broth culture showed differentiation patterns similar to those from swarming cells, suggesting that the agar surface culture may not be essential for the formation of differentiated cells.

  12. 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.

  13. Mixed culture polyhydroxyalkanoates production from sugar molasses: the use of a 2-stage CSTR system for culture selection.

    PubMed

    Albuquerque, M G E; Concas, S; Bengtsson, S; Reis, M A M

    2010-09-01

    Polyhydroxyalkanoates (PHAs) are promising biodegradable polymers. The use of mixed microbial cultures (MMC) and low cost feedstocks have a positive impact on the cost-effectiveness of the process. It has typically been carried out in Sequencing Batch Reactors (SBR). In this study, a 2-stage CSTR system (under Feast and Famine conditions) was used to effectively select for PHA-storing organisms using fermented molasses as feedstock. The effect of influent substrate concentration (60-120 Cmmol VFA/L) and HRT ratio between the reactors (0.2-0.5h/h) on the system's selection efficiency was assessed. It was shown that Feast reactor residual substrate concentration impacted on the selective pressure for PHA storage (due to substrate-dependent kinetic limitation). Moreover, a residual substrate concentration coming from the Feast to the Famine reactor did not jeopardize the physiological adaptation required for enhanced PHA storage. The culture reached a maximum PHA content of 61%. This success opens new perspectives to the use of wastewater treatment infrastructure for PHA production, thus valorizing either excess sludge or wastewaters.

  14. 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

  15. 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

  16. 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

  17. 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

  18. 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.

  19. Gibberellic acid production by free and immobilized cells in different culture systems.

    PubMed

    Durán-Páramo, Enrique; Molina-Jiménez, Héctor; Brito-Arias, Marco A; Robles-Martínez, Fabián

    2004-01-01

    Gibberellic acid production was studied in different fermentation systems. Free and immobilized cells of Gibberella fujikuroi cultures in shake-flask, stirred and fixed-bed reactors were evaluated for the production of gibberellic acid (GA3). Gibberellic acid production with free cells cultured in a stirred reactor reached 0.206 g/L and a yield of 0.078 g of GA3/g biomass.

  20. The fitness and functionality of culturally evolved communication systems.

    PubMed

    Fay, Nicolas; Garrod, Simon; Roberts, Leo

    2008-11-12

    This paper assesses whether human communication systems undergo the same progressive adaptation seen in animal communication systems and concrete artefacts. Four experiments compared the fitness of ad hoc sign systems created under different conditions when participants play a graphical communication task. Experiment 1 demonstrated that when participants are organized into interacting communities, a series of signs evolve that enhance individual learning and promote efficient decoding. No such benefits are found for signs that result from the local interactions of isolated pairs of interlocutors. Experiments 2 and 3 showed that the decoding benefits associated with community evolved signs cannot be attributed to superior sign encoding or detection. Experiment 4 revealed that naive overseers were better able to identify the meaning of community evolved signs when compared with isolated pair developed signs. Hence, the decoding benefits for community evolved signs arise from their greater residual iconicity. We argue that community evolved sign systems undergo a process of communicative selection and adaptation that promotes optimized sign systems. This results from the interplay between sign diversity and a global alignment constraint; pairwise interaction introduces a range of competing signs and the need to globally align on a single sign-meaning mapping for each referent applies selection pressure.

  1. The fitness and functionality of culturally evolved communication systems.

    PubMed

    Fay, Nicolas; Garrod, Simon; Roberts, Leo

    2008-11-12

    This paper assesses whether human communication systems undergo the same progressive adaptation seen in animal communication systems and concrete artefacts. Four experiments compared the fitness of ad hoc sign systems created under different conditions when participants play a graphical communication task. Experiment 1 demonstrated that when participants are organized into interacting communities, a series of signs evolve that enhance individual learning and promote efficient decoding. No such benefits are found for signs that result from the local interactions of isolated pairs of interlocutors. Experiments 2 and 3 showed that the decoding benefits associated with community evolved signs cannot be attributed to superior sign encoding or detection. Experiment 4 revealed that naive overseers were better able to identify the meaning of community evolved signs when compared with isolated pair developed signs. Hence, the decoding benefits for community evolved signs arise from their greater residual iconicity. We argue that community evolved sign systems undergo a process of communicative selection and adaptation that promotes optimized sign systems. This results from the interplay between sign diversity and a global alignment constraint; pairwise interaction introduces a range of competing signs and the need to globally align on a single sign-meaning mapping for each referent applies selection pressure. PMID:18799421

  2. 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.

  3. 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...

  4. 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…

  5. 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…

  6. 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…

  7. 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…

  8. 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

  9. 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

  10. Development of a definition, classification system, and model for cultural geology

    NASA Astrophysics Data System (ADS)

    Mitchell, Lloyd W., III

    The concept for this study is based upon a personal interest by the author, an American Indian, in promoting cultural perspectives in undergraduate college teaching and learning environments. Most academicians recognize that merged fields can enhance undergraduate curricula. However, conflict may occur when instructors attempt to merge social science fields such as history or philosophy with geoscience fields such as mining and geomorphology. For example, ideologies of Earth structures derived from scientific methodologies may conflict with historical and spiritual understandings of Earth structures held by American Indians. Specifically, this study addresses the problem of how to combine cultural studies with the geosciences into a new merged academic discipline called cultural geology. This study further attempts to develop the merged field of cultural geology using an approach consisting of three research foci: a definition, a classification system, and a model. Literature reviews were conducted for all three foci. Additionally, to better understand merged fields, a literature review was conducted specifically for academic fields that merged social and physical sciences. Methodologies concentrated on the three research foci: definition, classification system, and model. The definition was derived via a two-step process. The first step, developing keyword hierarchical ranking structures, was followed by creating and analyzing semantic word meaning lists. The classification system was developed by reviewing 102 classification systems and incorporating selected components into a system framework. The cultural geology model was created also utilizing a two-step process. A literature review of scientific models was conducted. Then, the definition and classification system were incorporated into a model felt to reflect the realm of cultural geology. A course syllabus was then developed that incorporated the resulting definition, classification system, and model. This

  11. Implementation of culturally targeted patient navigation system for screening colonoscopy in a direct referral system.

    PubMed

    Jandorf, Lina; Cooperman, Julia L; Stossel, Lauren M; Itzkowitz, Steven; Thompson, Hayley S; Villagra, Cristina; Thélémaque, Linda D; McGinn, Thomas; Winkel, Gary; Valdimarsdottir, Heiddis; Shelton, Rachel C; Redd, William

    2013-10-01

    Low-income minorities often face system-based and personal barriers to screening colonoscopy (SC). Culturally targeted patient navigation (CTPN) programs employing professional navigators (Pro-PNs) or community-based peer navigators (Peer-PNs) can help overcome barriers but are not widely implemented. In East Harlem, NY, USA, where approximately half the residents participate in SC, 315 African American patients referred for SC at a primary care clinic with a Direct Endoscopic Referral System were recruited between May 2008 and May 2010. After medical clearance, 240 were randomized to receive CTPN delivered by a Pro-PN (n = 106) or Peer-PN (n = 134). Successful navigation was measured by SC adherence rate, patient satisfaction and navigator trust. Study enrollment was 91.4% with no significant differences in SC adherence rates between Pro-PN (80.0%) and Peer-PN (71.3%) (P = 0.178). Participants in both groups reported high levels of satisfaction and trust. These findings suggest that CTPN Pro-PN and Peer-PN programs are effective in this urban primary care setting. We detail how we recruited and trained navigators, how CTPN was implemented and provide a preliminary answer to our questions of the study aims: can peer navigators be as effective as professionals and what is the potential impact of patient navigation on screening adherence?

  12. 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.

  13. 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...

  14. The rat whole embryo culture assay using the Dysmorphology Score system.

    PubMed

    Zhang, Cindy; Panzica-Kelly, Julie; Augustine-Rauch, Karen

    2013-01-01

    The rat whole embryo culture (WEC) system has been used extensively for characterizing teratogenic properties of test chemicals. In this chapter, we describe the methodology for culturing rat embryos as well as a new morphological score system, the Dysmorphology Score (DMS) system for assessing morphology of mid gestation (gestational day 11) rat embryos. In contrast to the developmental stage focused scoring associated with the Brown and Fabro score system, this new score system assesses the respective degree of severity of dysmorphology, which delineates normal from abnormal morphology of specific embryonic structures and organ systems. This score system generates an approach that allows rapid identification and quantification of adverse developmental findings, making it conducive for characterization of compounds for teratogenic properties and screening activities.

  15. An alternative long-term culture system for highly-pure mouse spermatogonial stem cells.

    PubMed

    He, Bao-Rong; Lu, Fan; Zhang, Lingling; Hao, Ding-Jun; Yang, Hao

    2015-06-01

    Increasing evidence suggests that spermatogonial stem cells (SSCs) have great clinical potential to give rise to a variety of cell types besides all spermatogenic lineage cells. The development of an efficient method for long-term culture of highly-pure SSCs is essential for further studies related to SSC biological events. Here, we describe an in vitro culture system obtaining mouse SSC cultures of high purity, viability, and proliferation. For establishing long-term cultures of SSCs, we mainly focused on isolation procedures and culture conditions. These included co-coating of extracellular substrates, that is, poly-L-lysine (PLL) and laminin, as well as combinatiorial use of three milder enzymes and simultaneously less trypsin to minimize enzyme-mediated degradation of SSCs. Furthermore, a unique purification procedure was performed to effectively eliminate contaminating non-SSCs. Finally, a critical step is to ensure SSC maintenance and expansion by utilizing optimal culture medium. Obtained data suggest that applying our optimally modified method, SSCs can be cultured for over 90 days with high purity (around 93.5%). Moreover, SSCs isolated and expanded using our protocol fulfills all criteria of SSCs without losing their stemness-characterized by SSC-phenotypic gene expression and long-term self-renewal. This study describes for the first time a protocol allowing isolation and expansion of SSCs suitable for numerous studies related to SSC-based clinical therapies of various diseases.

  16. Large-scale clinical comparison of the lysis-centrifugation and radiometric systems for blood culture

    SciTech Connect

    Brannon, P.; Kiehn, T.E.

    1985-12-01

    The Isolator 10 lysis-centrifugation blood culture system (E. I. du Pont de Nemours and Co., Inc., Wilmington, Del.) was compared with the BACTEC radiometric method (Johnston Laboratories, Inc., Towson, Md.) with 6B and 7D broth media for the recovery of bacteria and yeasts. From 11,000 blood cultures, 1,174 clinically significant organisms were isolated. The Isolator system recovered significantly more total organisms, members of the family Enterobacteriaceae, Staphylococcus spp., and yeasts. The BACTEC system recovered significantly more Pseudomonas spp., Streptococcus spp., and anaerobes. Of the Isolator colony counts, 87% measured less than 11 CFU/ml of blood. Organisms, on an average, were detected the same day from each of the two culture systems. Only 13 of the 975 BACTEC isolates (0.01%) were recovered by subculture of growth-index-negative bottles, and 12 of the 13 were detected in another broth blood culture taken within 24 h. Contaminants were recovered from 4.8% of the Isolator 10 and 2.3% of the BACTEC cultures.

  17. 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

  18. 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

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

    PubMed

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

    2016-05-15

    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.

  20. Interferon Response in Hepatitis C Virus (HCV) Infection: Lessons from Cell Culture Systems of HCV Infection.

    PubMed

    Sung, Pil Soo; Shin, Eui-Cheol; Yoon, Seung Kew

    2015-01-01

    Hepatitis C virus (HCV) is a positive-stranded RNA virus that infects approximately 130-170 million people worldwide. In 2005, the first HCV infection system in cell culture was established using clone JFH-1, which was isolated from a Japanese patient with fulminant HCV infection. JFH-1 replicates efficiently in hepatoma cells and infectious virion particles are released into the culture supernatant. The development of cell culture-derived HCV (HCVcc) systems has allowed us to understand how hosts respond to HCV infection and how HCV evades host responses. Although the mechanisms underlying the different outcomes of HCV infection are not fully understood, innate immune responses seem to have a critical impact on the outcome of HCV infection, as demonstrated by the prognostic value of IFN-λ gene polymorphisms among patients with chronic HCV infection. Herein, we review recent research on interferon response in HCV infection, particularly studies using HCVcc infection systems.

  1. 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

  2. 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.

  3. Consideration of the cellular microenvironment: physiologically relevant co-culture systems in drug discovery.

    PubMed

    L Berg, Ellen; Hsu, Yu-Chih; Lee, Jonathan A

    2014-04-01

    There is renewed interest in phenotypic approaches to drug discovery, using cell-based assays to select new drugs, with the goal of improving pharmaceutical success. Assays that are more predictive of human biology can help researchers achieve this goal. Primary cells are more physiologically relevant to human biology and advances are being made in methods to expand the available cell types and improve the potential clinical translation of these assays through the use of co-cultures or three-dimensional (3D) technologies. Of particular interest are assays that may be suitable for industrial scale drug discovery. Here we review the use of primary human cells and co-cultures in drug discovery and describe the characteristics of co-culture models for inflammation biology (BioMAP systems), neo-vascularization and tumor microenvironments. Finally we briefly describe technical trends that may enable and impact the development of physiologically relevant co-culture assays in the near future. PMID:24524933

  4. The response of mineralizing culture systems to microtextured and polished titanium surfaces.

    PubMed

    Hacking, S Adam; Harvey, Edward; Roughley, Peter; Tanzer, Michael; Bobyn, John

    2008-10-01

    The surface texture of titanium has a predictable effect on peri-implant tissue formation in vivo. When implanted in an osseous environment, smooth surfaces (R(a) < 0.5 mm) are generally apposed by fibrous tissue and textured surfaces (R(a) > 1.0 mm) are generally apposed by osseous tissue. Thus in vitro study assessed the mineralization and proliferation response of TF274, MC3T3-E1, murine femoral stromal cells and canine stromal cells to tissue culture plastic (R(a) = 0.001 mm), polished (R(a) = 0.01 mm) and irregularly textured (R(a) = 3.26 mm) titanium surfaces. Amongst all culture systems, proliferation was significantly decreased on textured vs. smooth surfaces. Midway through the culture of the canine marrow cells, the cell layer detached from the tissue culture plastic and polished titanium surfaces. The TF274, MC3T3-E1, murine femoral stromal cell systems formed a mineralized matrix on the tissue culture plastic and polished titanium surfaces which was not observed with the canine stromal cell system. Compared to the tissue culture plastic and polished titanium surfaces, matrix mineralization was significantly reduced on the textured titanium surfaces for the TF274, MC3T3-E1, murine femoral stromal systems, a result which was differed significantly in comparison to the canine stromal system. These results were surprising given the large number of reports concerning the in vivo response to titanium in clinical and pre-clinical studies. Further work is required to determine if the TF274, MC3T3-E1 and murine femoral stromal systems are suitable for the in vitro investigation of the effects of titanium surface texture on osteoblast activity.

  5. 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

  6. 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

  7. 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.

  8. 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.

  9. 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

  10. A 3D modeling and measurement system for cultural heritage preservation

    NASA Astrophysics Data System (ADS)

    Du, Guoguang; Zhou, Mingquan; Ren, Pu; Shui, Wuyang; Zhou, Pengbo; Wu, Zhongke

    2015-07-01

    Cultural Heritage reflects the human production, life style and environmental conditions of various historical periods. It exists as one of the major national carriers of national history and culture. In order to do better protection and utilization for these cultural heritages, a system of three-dimensional (3D) reconstruction and statistical measurement is proposed in this paper. The system solves the problems of cultural heritage's data storage, measurement and analysis. Firstly, for the high precision modeling and measurement problems, range data registration and integration algorithm used to achieve high precision 3D reconstruction. Secondly, multi-view stereo reconstruction method is used to solve the problem of rapid reconstruction by procedures such as the original image data pre-processing, camera calibration, point cloud modeling. At last, the artifacts' measure underlying database is established by calculating the measurements of the 3D model's surface. These measurements contain Euclidean distance between the points on the surface, geodesic distance between the points, normal and curvature in each point, superficial area of a region, volume of model's part and some other measurements. These measurements provide a basis for carrying out information mining of cultural heritage. The system has been applied to the applications of 3D modeling, data measurement of the Terracotta Warriors relics, Tibetan architecture and some other relics.

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

    PubMed

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

    2016-01-01

    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.

  12. Long-term culture system for selective growth of human B-cell progenitors.

    PubMed Central

    Rawlings, D J; Quan, S G; Kato, R M; Witte, O N

    1995-01-01

    We describe a simple reproducible system for enrichment and long-term culture of human B-cell progenitors. Enriched CD34+ cord blood mononuclear cells are seeded onto a murine stromal cell line to establish a biphasic culture system. These cultures are characterized by transient growth of myeloid cells followed by outgrowth of cells highly enriched for early B-cell progenitors. Cultures consisting of > 90% early B-lineage cells [expressing CD10, CD19, CD38, and CD45 but lacking CD20, CD22, CD23, and surface IgM] are maintained for > 12 weeks without growth factor addition. Cells remain predominantly germ line at the immunoglobulin locus and express only low levels of cytoplasmic mu chain, terminal deoxynucleotidyltransferase, and recombination-activating gene 1 product. They are unresponsive to the pre-B-cell growth factors interleukin 7 or stem cell factor, or both, suggesting that growth support is provided by a cross-reactive murine stromal cell factor. Cultured B-cell progenitors are generated in large numbers ( > 10(8) cells from a typical cord blood specimen) suitable for use in biochemical analysis and gene-transfer studies. This system should be useful for study of normal and abnormal early human B-lymphopoiesis. Images Fig. 2 Fig. 3 Fig. 4 PMID:7533295

  13. 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

  14. 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.

  15. 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

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

    PubMed Central

    Price, Travis K.; Dune, Tanaka; Hilt, Evann E.; Thomas-White, Krystal J.; Kliethermes, Stephanie; Brincat, Cynthia; Brubaker, Linda; Wolfe, Alan J.

    2016-01-01

    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

  17. Assessment of carotenoid production by Dunaliella salina in different culture systems and operation regimes.

    PubMed

    Prieto, Ana; Pedro Cañavate, J; García-González, Mercedes

    2011-01-20

    The effect of operation regime and culture system on carotenoid productivity by the halotolerant alga Dunaliella salina has been analyzed. Operation strategies tested included batch and semi continuous regime, as well as a two-stage approach run simultaneously in both, open tanks and closed reactor. The best results were obtained with the closed tubular photobioreactor. The highest carotenoid production (328.8 mg carotenoid l⁻¹ culture per month) was achieved with this culture system operated following the two-stage strategy. Also, closed tubular photobioreactor provided the highest carotenoid contents (10% of dry weight) in Dunaliella biomass and β-carotene abundance (90% of total carotenoids) as well as the highest 9-cis to all-trans β-carotene isomer ratio (1.5 at sunrise).

  18. 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

  19. Off-flavor characterization and depuration in Atlantic salmon cultured to food-size within closed-containment systems

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Atlantic salmon are typically cultured in marine net pens. However, technological advancements in recirculating aquaculture systems have increased the feasibility of culturing Atlantic salmon in land-based systems. One problem encountered when fish are harvested from recirculating systems is the pre...

  20. 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

  1. Comparison of defined culture systems for feeder cell free propagation of human embryonic stem cells

    PubMed Central

    Akopian, Veronika; Beil, Stephen; Benvenisty, Nissim; Brehm, Jennifer; Christie, Megan; Ford, Angela; Fox, Victoria; Gokhale, Paul J.; Healy, Lyn; Holm, Frida; Hovatta, Outi; Knowles, Barbara B.; Ludwig, Tenneille E.; McKay, Ronald D. G.; Miyazaki, Takamichi; Nakatsuji, Norio; Oh, Steve K. W.; Pera, Martin F.; Rossant, Janet; Stacey, Glyn N.; Suemori, Hirofumi

    2010-01-01

    There are many reports of defined culture systems for the propagation of human embryonic stem cells in the absence of feeder cell support, but no previous study has undertaken a multi-laboratory comparison of these diverse methodologies. In this study, five separate laboratories, each with experience in human embryonic stem cell culture, used a panel of ten embryonic stem cell lines (including WA09 as an index cell line common to all laboratories) to assess eight cell culture methods, with propagation in the presence of Knockout Serum Replacer, FGF-2, and mouse embryonic fibroblast feeder cell layers serving as a positive control. The cultures were assessed for up to ten passages for attachment, death, and differentiated morphology by phase contrast microscopy, for growth by serial cell counts, and for maintenance of stem cell surface marker expression by flow cytometry. Of the eight culture systems, only the control and those based on two commercial media, mTeSR1 and STEMPRO, supported maintenance of most cell lines for ten passages. Cultures grown in the remaining media failed before this point due to lack of attachment, cell death, or overt cell differentiation. Possible explanations for relative success of the commercial formulations in this study, and the lack of success with other formulations from academic groups compared to previously published results, include: the complex combination of growth factors present in the commercial preparations; improved development, manufacture, and quality control in the commercial products; differences in epigenetic adaptation to culture in vitro between different ES cell lines grown in different laboratories. PMID:20186512

  2. 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

  3. 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

  4. Mini-scale bioprocessing systems for highly parallel animal cell cultures.

    PubMed

    Kim, Beum Jun; Diao, Jinpian; Shuler, Michael L

    2012-01-01

    Animal cells have been used extensively in therapeutic protein production. The growth of animal cells and the expression of therapeutic proteins are highly dependent on the culturing environments. A large number of experimental permutations need to be explored to identify the optimal culturing conditions. Miniaturized bioreactors are well suited for such tasks as they offer high-throughput parallel operation and reduce cost of reagents. They can also be automated and be coupled to downstream analytical units for online measurements of culture products. This review summarizes the current status of miniaturized bioreactors for animal cell cultivation based on the design categories: microtiter plates, flasks, stirred tank reactors, novel designs with active mixing, and microfluidic cell culture devices. We compare cell density and product titer, for batch or fed-batch modes for each system. Monitoring/controlling devices for engineering parameters such as pH, dissolved oxygen, and dissolved carbon dioxide, which could be applied to such systems, are summarized. Finally, mini-scale tools for process performance evaluation for animal cell cultures are discussed: total cell density, cell viability, product titer and quality, substrates, and metabolites profiles. PMID:22522970

  5. 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.

  6. The Problem of Cultural Fit--What Can We Learn from Borrowing the German Dual System?

    ERIC Educational Resources Information Center

    Lewis, Theodore

    2007-01-01

    Culture impedes educational transactions between developed countries, and is often an area of tension when developing countries try to borrow models, or have models imposed upon them. This article uses the German Dual System as a case for examining issues surrounding the adaptation of borrowed models. Since attempts at borrowing models can fail…

  7. 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,…

  8. 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…

  9. 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…

  10. 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…

  11. 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…

  12. 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…

  13. 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,…

  14. 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...

  15. 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…

  16. 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…

  17. 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…

  18. Cultural Competence, Systems of Care, and Students with Emotional and Behavioral Challenges

    ERIC Educational Resources Information Center

    Meyer, Lakeisha D.; Anderson, Jeffrey A.; McQueen, Kand

    2013-01-01

    For youth with emotional and behavioral challenges, there is a trend of minority overrepresentation in restrictive settings. "System of care" refers to an approach that emphasizes cultural competence and providing services in the least restrictive setting. This study investigated the relationship between placement restrictiveness,…

  19. 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…

  20. 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

  1. 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

  2. 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.

  3. 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.

  4. An aeroponic culture system for the study of root herbivory on Arabidopsis thaliana

    PubMed Central

    2011-01-01

    Background Plant defense against herbivory has been studied primarily in aerial tissues. However, complex defense mechanisms have evolved in all parts of the plant to combat herbivore attack and these mechanisms are likely to differ in the aerial and subterranean environment. Research investigating defense responses belowground has been hindered by experimental difficulties associated with the accessibility and quality of root tissue and the lack of bioassays using model plants with altered defense profiles. Results We have developed an aeroponic culture system based on a calcined clay substrate that allows insect herbivores to feed on plant roots while providing easy recovery of the root tissue. The culture method was validated by a root-herbivore system developed for Arabidopsis thaliana and the herbivore Bradysia spp. (fungus gnat). Arabidopsis root mass obtained from aeroponically grown plants was comparable to that from other culture systems, and the plants were morphologically normal. Bradysia larvae caused considerable root damage resulting in reduced root biomass and water absorption. After feeding on the aeroponically grown root tissue, the larvae pupated and emerged as adults. Root damage of mature plants cultivated in aeroponic substrate was compared to that of Arabidopsis seedlings grown in potting mix. Seedlings were notably more susceptible to Bradysia feeding than mature plants and showed decreased overall growth and survival rates. Conclusions A root-herbivore system consisting of Arabidopsis thaliana and larvae of the opportunistic herbivore Bradysia spp. has been established that mimics herbivory in the rhizosphere. Bradysia infestation of Arabidopsis grown in this culture system significantly affects plant performance. The culture method will allow simple profiling and in vivo functional analysis of root defenses such as chemical defense metabolites that are released in response to belowground insect attack. PMID:21392399

  5. 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%.

  6. Reproducibility of CSF quantitative culture methods for estimating rate of clearance in cryptococcal meningitis.

    PubMed

    Dyal, Jonathan; Akampurira, Andrew; Rhein, Joshua; Morawski, Bozena M; Kiggundu, Reuben; Nabeta, Henry W; Musubire, Abdu K; Bahr, Nathan C; Williams, Darlisha A; Bicanic, Tihana; Larsen, Robert A; Meya, David B; Boulware, David R

    2016-05-01

    Quantitative cerebrospinal fluid (CSF) cultures provide a measure of disease severity in cryptococcal meningitis. The fungal clearance rate by quantitative cultures has become a primary endpoint for phase II clinical trials. This study determined the inter-assay accuracy of three different quantitative culture methodologies. Among 91 participants with meningitis symptoms in Kampala, Uganda, during August-November 2013, 305 CSF samples were prospectively collected from patients at multiple time points during treatment. Samples were simultaneously cultured by three methods: (1) St. George's 100 mcl input volume of CSF with five 1:10 serial dilutions, (2) AIDS Clinical Trials Group (ACTG) method using 1000, 100, 10 mcl input volumes, and two 1:100 dilutions with 100 and 10 mcl input volume per dilution on seven agar plates; and (3) 10 mcl calibrated loop of undiluted and 1:100 diluted CSF (loop). Quantitative culture values did not statistically differ between St. George-ACTG methods (P= .09) but did for St. George-10 mcl loop (P< .001). Repeated measures pairwise correlation between any of the methods was high (r≥0.88). For detecting sterility, the ACTG-method had the highest negative predictive value of 97% (91% St. George, 60% loop), but the ACTG-method had occasional (∼10%) difficulties in quantification due to colony clumping. For CSF clearance rate, St. George-ACTG methods did not differ overall (mean -0.05 ± 0.07 log10CFU/ml/day;P= .14) on a group level; however, individual-level clearance varied. The St. George and ACTG quantitative CSF culture methods produced comparable but not identical results. Quantitative cultures can inform treatment management strategies.

  7. Health system factors affecting communication with pediatricians: gendered work culture in primary care.

    PubMed

    Lynch, Sean

    2011-01-01

    This qualitative study examined the roles that practice setting, education level, and gender may play in social workers' communication satisfaction with pediatricians. Taking an ethnographic approach, the researcher interviewed social workers and pediatricians who worked together to provide mental health services in primary care. The results suggested that gender at the health system level may be an issue and that gendered work culture in primary care was a factor in communication. In particular, reimbursement, an aspect of the gendered work culture, was a substantial communication barrier, and the implications for Medicaid billing are discussed. PMID:22085327

  8. 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.

  9. 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

  10. 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

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

    PubMed Central

    Zhao, Yang; Liu, Xiao-guang; Shi, Hao-qing; Hu, Ke-yao; Zhu, Guan-qun; Ma, Bo; Niu, Hai-tao

    2015-01-01

    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

  12. 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

  13. 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.

  14. B35 neuroblastoma cells: an easily transfected, cultured cell model of central nervous system neurons.

    PubMed

    Otey, Carol A; Boukhelifa, Malika; Maness, Patricia

    2003-01-01

    A panel of neuronal cell lines was derived from tumors of the neonatal rat central nervous system (CNS) in 1974, and two of these lines are in wide use today. Both the B35 and B50 lines offer a number of advantages to researchers who study CNS neurons in culture: they are simple to grow, to differentiate, and to transfect. B50 cells have been used extensively in the study of neuronal cell death, toxicology, and differentiation, whereas B35 cells have proven useful in the molecular analysis of endocytosis and of signaling pathways, in particular those that guide axonal outgrowth and cell motility. This chapter provides protocols for growing and transfecting B35 cells, selecting stable transfectants, exploring protein function using an antisense approach, and assaying cell motility in a Transwell chamber. All of these protocols have been written for researchers who have some skill in basic cell culture techniques, but previous experience with cultured neurons is not required.

  15. Biodegradation of haloacetic acids by bacterial isolates and enrichment cultures from drinking water systems.

    PubMed

    Zhang, Ping; Lapara, Timothy M; Goslan, Emma H; Xie, Yuefeng; Parsons, Simon A; Hozalski, Raymond M

    2009-05-01

    Biodegradation is a potentially important loss process for haloacetic acids (HAAs), a class of chlorination byproducts, in water treatment and distribution systems, but little is known about the organisms involved (i.e., identity, substrate range, biodegradation kinetics). In this research, 10 biomass samples (i.e., tap water, distribution system biofilms, and prechlorinated granular activated carbon filters) from nine drinking water systems were used to inoculate a total of thirty enrichment cultures fed monochloroacetic acid (MCAA), dichloroacetic acid (DCAA), or trichloroacetic (TCAA) as sole carbon and energy source. HAA degraders were successfully enriched from the biofilm samples (GAC and distribution system) but rarely from tap water. Half of the MCAA and DCAA enrichment cultures were positive, whereas only one TCAA culture was positive (two were inconclusive). Eight unique HAA-degrading isolates were obtained including several Afipia spp. and a Methylobacterium sp.; all isolates were members of the phylum Proteobacteria. MCAA, monobromoacetic acid (MBAA), and monoiodoacetic acid (MIAA) were rapidly degraded by all isolates, and DCAA and tribromoacetic (TBAA) were also relatively labile. TCAA and dibromoacetic acid (DBAA)were degraded by only three isolates and degradation lagged behind the other HAAs. Detailed DCAA biodegradation kinetics were obtained for two selected isolates and two enrichment cultures. The maximum biomass-normalized degradation rates (Vm) were 0.27 and 0.97 microg DCAA/ microg protein/h for Methylobacterium fujisawaense strain PAWDI and Afipia felis strain EMD2, respectively, which were comparable to the values obtained for the enrichment cultures from which those organisms were isolated (0.39 and 1.37 microg DCAN/microg protein/h, respectively). The half-saturation constant (Km) values ranged from 4.38 to 77.91 microg DCAA/L and the cell yields ranged from 14.4 to 36.1 mg protein/g DCAA.

  16. [Identification of staphylococci directly from positive blood culture bottles by MALDI-TOF MS system].

    PubMed

    Kilic, Abdullah; Baysallar, Mehmet

    2014-07-01

    Bloodstream infections are substantial causes of morbidity and mortality worldwide. Staphylococcus species are the most commonly isolated microorganisms from blood cultures in clinical microbiology laboratories. MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization Time- of- Flight Mass Spectrometry) system allows the identification of microorganisms directly from positive blood culture bottles. The aim of this study was to evaluate the MALDI-TOF MS system for the identification of staphylococci directly from the positive blood culture bottles which revealed the presence of gram-positive cocci by staining. A total of 96 positive blood culture bottles that yielded gram-positive cocci by Gram stain were evaluated. These blood cultures were obtained from 69 patients between December 2013-February 2014. Conventional methods and BD Phoenix™ automated bacterial identification system (Becton Dickinson, USA) were used for routine identification. The strains were also identified by real-time Taqman PCR (qPCR) which was considered as the reference method. In MALDI-TOF MS method, MALDI Sepsityper™ Kit was used for the bacterial identification and all measurements were carried out by using Microflex LT instrument and FlexControl 3.0 software (Bruker Daltonics, USA). Of 96 culture bottles positive for gram-positive cocci, 90 were correctly identified as staphylococci at genus level with all the three study methods (qPCR, BD Phoenix, Bruker MALDI-TOF MS). The other six samples were identified as Enterococcus faecium (n= 4) and Streptococcus pyogenes (n= 2) by both Phoenix and the MALDI-TOF systems. Of the 90 samples, 87 were identified at the species level (15 S.aureus, 33 S.epidermidis, 29 S.hominis, 10 S.haemolyticus) and three at the genus level by the reference qPCR method. When comparing the results obtained by qPCR and Bruker MALDI-TOF MS, incompatibility was detected for three isolates. Those isolates were identified as S.hominis by qPCR, however two of them were

  17. [Identification of staphylococci directly from positive blood culture bottles by MALDI-TOF MS system].

    PubMed

    Kilic, Abdullah; Baysallar, Mehmet

    2014-07-01

    Bloodstream infections are substantial causes of morbidity and mortality worldwide. Staphylococcus species are the most commonly isolated microorganisms from blood cultures in clinical microbiology laboratories. MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization Time- of- Flight Mass Spectrometry) system allows the identification of microorganisms directly from positive blood culture bottles. The aim of this study was to evaluate the MALDI-TOF MS system for the identification of staphylococci directly from the positive blood culture bottles which revealed the presence of gram-positive cocci by staining. A total of 96 positive blood culture bottles that yielded gram-positive cocci by Gram stain were evaluated. These blood cultures were obtained from 69 patients between December 2013-February 2014. Conventional methods and BD Phoenix™ automated bacterial identification system (Becton Dickinson, USA) were used for routine identification. The strains were also identified by real-time Taqman PCR (qPCR) which was considered as the reference method. In MALDI-TOF MS method, MALDI Sepsityper™ Kit was used for the bacterial identification and all measurements were carried out by using Microflex LT instrument and FlexControl 3.0 software (Bruker Daltonics, USA). Of 96 culture bottles positive for gram-positive cocci, 90 were correctly identified as staphylococci at genus level with all the three study methods (qPCR, BD Phoenix, Bruker MALDI-TOF MS). The other six samples were identified as Enterococcus faecium (n= 4) and Streptococcus pyogenes (n= 2) by both Phoenix and the MALDI-TOF systems. Of the 90 samples, 87 were identified at the species level (15 S.aureus, 33 S.epidermidis, 29 S.hominis, 10 S.haemolyticus) and three at the genus level by the reference qPCR method. When comparing the results obtained by qPCR and Bruker MALDI-TOF MS, incompatibility was detected for three isolates. Those isolates were identified as S.hominis by qPCR, however two of them were

  18. Plasma-mediated inactivation of Pseudomonas aeruginosa biofilms grown on borosilicate surfaces under continuous culture system.

    PubMed

    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 culturability 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

  19. 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

  20. 3d Modeling of cultural heritage objects with a structured light system.

    NASA Astrophysics Data System (ADS)

    Akca, Devrim

    3D modeling of cultural heritage objects is an expanding application area. The selection of the right technology is very important and strictly related to the project requirements, budget and user's experience. The triangulation based active sensors, e.g. structured light systems are used for many kinds of 3D object reconstruction tasks and in particular for 3D recording of cultural heritage objects. This study presents the experiences in the results of two such projects in which a close-range structured light system is used for the 3D digitization. The paper includes the essential steps of the 3D object modeling pipeline, i.e. digitization, registration, surface triangulation, editing, texture mapping and visualization. The capabilities of the used hardware and software are addressed. Particular emphasis is given to a coded structured light system as an option for data acquisition.

  1. 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.; Gorsuch, Joseph W.; Lower, William R.; Wang, Wun-cheng; Lewis, M.A.

    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.

  2. Effect of Culture System on Developmental Competence, Cryosurvival and DNA-Fragmentation of In Vitro Bovine Blastocysts

    PubMed Central

    Hajian, Mahdi; Hosseini, Seyed Morteza; Asgari, Vajiheh; Ostadhoosseini, Somayyeh; Forouzanfar, Mohsen; Nasr Esfahani, Mohammad Hossein

    2011-01-01

    Background This study investigated the effect of two in vitro embryo culture systems (co-culture system versus cell-free sequential-media) on developmental competence, cryosurvival and DNA- fragmentation of in vitro developed bovine blastocysts. Materials and Methods Bovine presumptive zygotes were cultured in Ménézo's B2 (B2) plus vero-cells or sequential synthetic oviductal fluid (SOF) for eight days. Subsequently, half of the expanded blastocysts developed in both groups were vitrified, warmed within 30 minutes and post- warming embryos along with their corresponding non-vitrified embryos were cultured for two additional days in the same medium used before vitrification. Embryo development, cryosurvival and apoptosis were compared between the groups. Results For non-vitrified embryos, culture in SOF significantly promoted the potency of embryos to develop into blastocysts compared with the co-culture system. The difference in post vitrification survival rate of SOF blastocysts (83.3%) was insignificant compared with co-culture (84.3%). However, while total cell number of warmed blastocysts in the co-culture system was significantly higher in the co-culture versus the sequential system (215.4 vs. 170.4), the quality of survived embryos in terms of hatching ability and apoptosis was adversely affected by co-culture compared with SOF (65.0% vs. 74.3%, and 13.5% vs. 10.0%, respectively; p<0.05). Conclusion Although co-culture system may increase the viability of embryos following cryopreservation, the potency and dynamics of blastocyst formation significantly increased with sequential media compared to the co-culture system which can compensate for the lower efficiency of sequential media for vitrification/warming purposes. PMID:24917920

  3. 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…

  4. Testing a theory of organizational culture, climate and youth outcomes in child welfare systems: a United States national study.

    PubMed

    Williams, Nathaniel J; Glisson, Charles

    2014-04-01

    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.

  5. 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

  6. Organotypic cultures as tool to test long-term effects of chemicals on the nervous system.

    PubMed

    Peña, F

    2010-01-01

    The study of neuroscience has vastly benefited from the use of brain slices. This preparation has been fundamental for the understanding of the cellular basis of nervous system function as well as for the study of the mechanisms involved in neuronal network dysfunction. This experimental model provides flexible access, and control of, specific neural circuits and maintains their basic properties, allowing them to reproduce most of their natural network activities. Brain slices permit the combination of sophisticated techniques such as electrophysiology, fluorescence imaging, pharmacology, molecular biology, etc. More recently, the development of organotypic brain slice cultures has expanded the use of modern technical approaches to the study neuronal networks, while increasing their possibilities of evaluating long-term effects of acute experimental conditions, as well as the effects of chronic treatments on neuronal network function in vitro. Here, I will provide an overview of the use of organotypic cultures to understand neuronal network function and dysfunction, as well as the pharmacological approaches used for these studies. As a final example, I will review the studies performed in organotypic cultures regarding the deleterious effects of long-term amyloid beta application on neuronal networks in vitro, as well as the use of drugs that may prevent or revert their deleterious effects on nervous system function. Overall, this review will provide elements to support the use of organotypic cultures as a very reliable model to explore long-term neuropharmacological studies in vitro. PMID:20156165

  7. 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

  8. 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

  9. Evaluation of isolator system and large-volume centrifugation method for culturing body fluids.

    PubMed Central

    Elston, H R; Wang, M; Philip, A

    1990-01-01

    The Isolator system was compared with the large-volume centrifugation method for processing and recovering organisms from body fluids other than blood, cerebrospinal fluid, and urine. A total of 155 body fluid samples were processed for the recovery of clinically significant organisms. Of the 55 positive cultures, Isolator detected 94% and the large-volume centrifugation method detected 64%. The time necessary to indicate positivity was not significantly different in the two methods; however, in five cases, the Isolator system yielded clinically significant organisms 24 h sooner than the conventional method. The Isolator system was found to be a more sensitive alternative than the conventional large-volume centrifugation method. PMID:2405006

  10. 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

  11. 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

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

    PubMed

    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. PMID:26729451

  13. 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.

  14. Bioreactors to Influence Stem Cell Fate: Augmentation of Mesenchymal Stem Cell Signaling Pathways via Dynamic Culture Systems

    PubMed Central

    Yeatts, Andrew B.; Choquette, Daniel T.; Fisher, John P.

    2012-01-01

    Background Mesenchymal stem cells (MSCs) are a promising cell source for bone and cartilage tissue engineering as they can be easily isolated from the body and differentiated into osteoblasts and chondrocytes. A cell based tissue engineering strategy using MSCs often involves the culture of these cells on three-dimensional scaffolds; however the size of these scaffolds and the cell population they can support can be restricted in traditional static culture. Thus dynamic culture in bioreactor systems provides a promising means to culture and differentiate MSCs in vitro. Scope of Review This review seeks to characterize key MSC differentiation signaling pathways and provides evidence as to how dynamic culture is augmenting these pathways. Following an overview of dynamic culture systems, discussion will be provided on how these systems can effectively modify and maintain important culture parameters including oxygen content and shear stress. Literature is reviewed for both a highlight of key signaling pathways and evidence for regulation of these signaling pathways via dynamic culture systems. Major Conclusions The ability to understand how these culture systems are affecting MSC signaling pathways could lead to a shear or oxygen regime to direct stem cell differentiation. In this way the efficacy of in vitro culture and differentiation of MSCs on three-dimensional scaffolds could be greatly increased. General Significance Bioreactor systems have the ability to control many key differentiation stimuli including mechanical stress and oxygen content. The further integration of cell signaling investigations within dynamic culture systems will lead to a quicker realization of the promise of tissue engineering and regenerative medicine. PMID:22705676

  15. 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

  16. Soft material-based microculture system having air permeable cover sheet for the protoplast culture of Nicotiana tabacum.

    PubMed

    Ju, Jong Il; Ko, Jung-Moon; Kim, So Hyeon; Baek, Ju Yeoul; Cha, Hyeon-Cheol; Lee, Sang Hoon

    2006-08-01

    In plant cell culture, the delivery of nutrition and gas (mainly oxygen) to the cells is the most important factor for viability. In this paper, we propose a polydimethylsiloxane (PDMS)-based microculture system that is designed to have good aeration. PDMS is known to have excellent air permeability, and through the experimental method, we investigated the relation between the degree of air delivery and the thickness of the PDMS sheet covering the culture chamber. We determined the proper thickness of the cover sheet, and cultured protoplasts of Nicotiana tabacum in a culture chamber covered with a PDMS sheet having thickness of 400 microm. The cells were successfully divided, and lived well inside the culture chamber for 10 days. In addition, protoplasts were cultured inside the culture chambers covered with the cover glass and the PDMS sheet, respectively, and the microcolonies were formed well inside the PDMS covered chamber after 10 days.

  17. Review of vascularised bone tissue-engineering strategies with a focus on co-culture systems.

    PubMed

    Liu, Yuchun; Chan, Jerry K Y; Teoh, Swee-Hin

    2015-02-01

    Poor angiogenesis within tissue-engineered grafts has been identified as a main challenge limiting the clinical introduction of bone tissue-engineering (BTE) approaches for the repair of large bone defects. Thick BTE grafts often exhibit poor cellular viability particularly at the core, leading to graft failure and lack of integration with host tissues. Various BTE approaches have been explored for improving vascularisation in tissue-engineered constructs and are briefly discussed in this review. Recent investigations relating to co-culture systems of endothelial and osteoblast-like cells have shown evidence of BTE efficacy in increasing vascularization in thick constructs. This review provides an overview of key concepts related to bone formation and then focuses on the current state of engineered vascularized co-culture systems using bone repair as a model. It will also address key questions regarding the generation of clinically relevant vascularized bone constructs as well as potential directions and considerations for research with the objective of pursuing engineered co-culture systems in other disciplines of vascularized regenerative medicine. The final objective is to generate serious and functional long-lasting vessels for sustainable angiogenesis that will enable enhanced cellular survival within thick voluminous bone grafts, thereby aiding in bone formation and remodelling in the long term. However, more evidence about the quality of blood vessels formed and its associated functional improvement in bone formation as well as a mechanistic understanding of their interactions are necessary for designing better therapeutic strategies for translation to clinical settings.

  18. 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.

  19. 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.

  20. The development of an in-vitro perfusion system for studies on cultured cells.

    PubMed Central

    Salih, V.; Greenwald, S. E.; Chong, C. F.; Coumbe, A.; Berry, C. L.

    1992-01-01

    The design and use of a perfusion system, using a modified flow chamber for studies on cultured animal cells, is described. Rat thoracic aorta smooth muscle cells were isolated by an explant method and grown on Thermanox coverslips. These were introduced into the flow chamber. A flow rate of 25ml/min and a shear stress of 14.6 dynes/cm2 (12 dyne = 10 microN) (both within physiological limits) were maintained. Cells remained attached to the coverslips after 8h of perfusion with culture medium. The effect of exposing rat smooth muscle cells to the cardiovascular toxin, allylamine, is also described. The components of the system are routinely available, simple to clean, easy to assemble and sterilize. The incorporation of an in-line sensor that monitors pH, PO2, PCO2 and temperature ensures that the perfusion conditions remain within physiological limits. Automation means that minimal supervision is required. This system provides a potential mechanism in which cultured vascular cells may be perfused under controlled haemodynamic conditions, and their response to a cytotoxin may be evaluated. Images Fig. 3 Fig. 4 Fig. 5 PMID:1419778

  1. [Establishment of culture system of Silybum marianum hairy roots and determination of silybin].

    PubMed

    Zhang, Shu-Li; Zhang, Tian-Zhu; Yang, Shi-Hai

    2014-06-01

    This research uses six Agrobacterium rhizogenes R1601, R15384, R1000, A4, R1025 and R1 to infect silymarin explants to induce hairy roots and silibin. All of the six A. rhizogenes can induce Silybum marianum to generate hairy roots and the A. rhizogene A4 shows comparatively high infection on the plant. This research determines the condition to induce silymarin hairy roots by the factors of infection time, pre-culturing, co-culturing and pH value. The fact that MS liquid medium fits the proliferation of silymarin hairy roots is determined. Through PCR molecular identification, it can be seen that the DNA plasmids in the A. rhizogenes are successfully integrated into the genome of transformed roots. Using liquid chromatography, it is determined that the silibin content in silymarin hairy roots is 2.5 times that in the plant In this research, the silymarin hairy roots culturing system is established, which lays a foundation for the study of culturing silymarin hairy roots and producing silibin.

  2. 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.

  3. Adipose progenitor cells reside among the mature adipocytes: morphological research using an organotypic culture system.

    PubMed

    Anayama, Hisashi; Fukuda, Ryo; Yamate, Jyoji

    2015-11-01

    The precise localization and biological characteristics of the adipose progenitor cells are still a focus of debate. In this study, the localization of the adipose progenitor cells was determined using an organotypic culture system of adipose tissue slices. The tissue slices of subcutaneous white adipose tissue from rats were placed on a porous membrane and cultured at the interface between air and the culture medium for up to 5 days with or without adipogenic stimulation. The structure of adipose tissue components was sufficiently preserved during the culture and, following adipogenic stimulation with insulin, dexamethasone, and 3-isobutyl-1-methylxanthine, numerous multilocular adipocytes appeared in the interstitium among the mature adipocytes. Histomorphological 3-D observation using confocal laser microscopy revealed the presence of small mesenchymal cells containing little or no fat residing in the perivascular region and on the mature adipocytes and differentiation from the pre-existing mesenchymal cells to multilocular adipocytes. Immunohistochemistry demonstrated that these cells were initially present within the fibronectin-positive extracellular matrix (ECM). The adipose differentiation of the mesenchymal cells was confirmed by the enhanced expression of C/EBP-β suggesting adipose differentiation and the concurrent advent of CD105-expressing mesenchymal cells within the interstitium of the mature adipocytes. Based on the above, the mesenchymal cells embedded in the ECM around the mature adipocytes were confirmed to be responsible for adipogenesis because the transition of the mesenchymal cells to the stem state contributed to the increase in the number of adipocytes in rat adipose tissue.

  4. 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

  5. A sandwich-cultured rat hepatocyte system with increased metabolic competence evaluated by gene expression profiling.

    PubMed

    Kienhuis, A S; Wortelboer, H M; Maas, W J; van Herwijnen, M; Kleinjans, J C S; van Delft, J H M; Stierum, R H

    2007-08-01

    A rapid decline of cytochrome P450 (CYP450) enzyme activities remains a drawback of rat hepatocyte-based in vitro cultures. Consequently, judgment of the toxic potential of compounds that need bioactivation by CYP450s may not be adequate using this model. In the present study, an improved hepatocyte-based in vitro system was developed with special focus on metabolic competence. Therefore, a mixture of CYP450 inducers, phenobarbital, dexamethasone and beta-naphthoflavone, was added to culture medium of sandwich-cultured rat hepatocytes. The resulting modified model was evaluated by comparing its genome-wide expression profiles with liver and a standard model without the inducer mixture. Metabolic capacity for CYP450 enzymes showed that the modified model resembled more closely the in vivo situation. Gene expression results revealed large differences between in vivo and both in vitro models. The slight differences between the two sandwich models were predominantly represented by gene expression changes in CYP450s. Importantly, in the modified model, expression ratios of the phase I and the majority of phase II genes more closely resembled liver in vivo. The CYP450 enzyme activities corresponded with gene expression data. In conclusion, for toxicological applications using sandwich-cultured hepatocytes, the modified model may be preferred. PMID:17336492

  6. New directions in capacity building: incorporating cultural competence into the interactive systems framework.

    PubMed

    Gregory, Henry; Van Orden, Onna; Jordan, Lisa; Portnoy, Galina A; Welsh, Elena; Betkowski, Jennifer; Charles, Jade Wolfman; DiClemente, Carlo C

    2012-12-01

    The UMBC Psychology Department's Center for Community Collaboration (CCC) provides training and support for capacity building to promote substance abuse and mental health treatment as well as adherence improvement in community agencies funded through the Ryan White Act serving persons living with HIV/AIDS. This article describes an approach to dissemination of Evidence Based Practices (EBPs) for these services that uses the Interactive Systems Framework (ISF) and incorporates a collaborative process involving trainer cultural competence, along with a comprehensive assessment of organizational needs, culture, and climate that culminates in tailored training and ongoing collaboration. This article provides: (1) an overview of the CCC's expanded ISF for the effective dissemination of two EBPs-motivational interviewing and the stages of change perspective; (2) an examination of the role of trainer cultural competence within the ISF framework, particularly attending to organizational culture and climate; and (3) case examples to demonstrate this approach for both general and innovation-specific capacity building in two community based organizations.

  7. Inhibitory Effect of Progesterone on Cervical Tissue Formation in a Three-Dimensional Culture System with Human Cervical Fibroblasts1

    PubMed Central

    House, Michael; Tadesse-Telila, Serkalem; Norwitz, Errol R.; Socrate, Simona; Kaplan, David L.

    2013-01-01

    ABSTRACT Progesterone supplementation is recommended to prevent preterm birth in women with a short cervix, but the mechanism is unclear. We hypothesize that progesterone acts by altering the composition of the cervical extracellular matrix (ECM). We tested this hypothesis using human cervical fibroblasts in both two-dimensional (2D) and three-dimensional (3D) cultures. For 2D culture, cells were seeded in 6-well plates and cultured with media supplemented with estradiol (10−8 M), progesterone (10−7 or 10−6 M), and vehicle. For 3D culture, the cells were cultured on a porous silk protein scaffold system. Progesterone and estrogen receptors were documented by immunohistochemistry and Western blot analysis. In both 2D and 3D cultures, decreased collagen synthesis was seen with increased progesterone concentration. Three-dimensional cultures could be maintained significantly longer than 2D cultures, and the morphology of 3D cultures appeared similar to native cervical tissue. Thus, further studies were performed in 3D culture. To determine the effect of progesterone concentration, the 3D scaffolds were cultured with estradiol (10−8 M) and five conditions: vehicle; 10−9, 10−8, or 10−7 M progesterone; or 10−7 M progesterone plus 10−6 M mifepristone. The highest progesterone concentration correlated with the least amount of collagen synthesis. Collagen synthesis progressively increased as progesterone concentration decreased. This effect was partially antagonized by mifepristone, suggesting the mechanism is mediated by the progesterone receptor. This hormonally responsive 3D culture system supports the hypothesis that progesterone has a direct effect on remodeling cervical ECM during pregnancy. The 3D culture system could be useful for studying the mechanism of progesterone effects on the cervix. PMID:24285720

  8. 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

  9. Validation of the BacT/ALERT®3D automated culture system for the detection of microbial contamination of epithelial cell culture medium.

    PubMed

    Plantamura, E; Huyghe, G; Panterne, B; Delesalle, N; Thépot, A; Reverdy, M E; Damour, O; Auxenfans, Céline

    2012-08-01

    Living tissue engineering for regenerative therapy cannot withstand the usual pharmacopoeia methods of purification and terminal sterilization. Consequently, these products must be manufactured under aseptic conditions at microbiologically controlled environment facilities. This study was proposed to validate BacT/ALERT(®)3D automated culture system for microbiological control of epithelial cell culture medium (ECCM). Suspensions of the nine microorganisms recommended by the European Pharmacopoeia (Chap. 2.6.27: "Microbiological control of cellular products"), plus one species from oral mucosa and two negative controls with no microorganisms were prepared in ECCM. They were inoculated in FA (anaerobic) and SN (aerobic) culture bottles (Biomérieux, Lyon, France) and incubated in a BacT/ALERT(®)3D automated culture system. For each species, five sets of bottles were inoculated for reproducibility testing: one sample was incubated at the French Health Products Agency laboratory (reference) and the four others at Cell and Tissue Bank of Lyon, France. The specificity of the positive culture bottles was verified by Gram staining and then subcultured to identify the microorganism grown. The BacT/ALERT(®)3D system detected all the inoculated microorganisms in less than 2 days except Propionibacterium acnes which was detected in 3 days. In conclusion, this study demonstrates that the BacT/ALERT(®)3D system can detect both aerobic and anaerobic bacterial and fungal contamination of an epithelial cell culture medium consistent with the European Pharmacopoeia chapter 2.6.27 recommendations. It showed the specificity, sensitivity, and precision of the BacT/ALERT(®)3D method, since all the microorganisms seeded were detected in both sites and the uncontaminated medium ECCM remained negative at 7 days. PMID:22160810

  10. Investigation of the interaction of cardiotoxic anticancer agents using the fetal mouse heart organ culture system

    SciTech Connect

    Kimler, B.F.; Rethorst, R.D.; Cox, G.G.

    1986-01-01

    The fetal mouse heart organ culture system was utilized in an effort to document and predict the potential cardiotoxic effects of ionizing radiation, Adriamycin (ADR), and Dihydroxyanthraquinone (DHAQ); alone and in combination. These antineoplastic agents have been shown to produce clinical cardiomyopathy which is often dose-limiting. Fetal mouse hearts (gestational day 17) were removed and placed in a culture system of 6-well microtiter plates. A single heart was placed in each well on a piece of aluminium mesh, above the culture medium but bathed by capillary action. The plates were then placed in a 100% oxygen environment and incubated at 37/sup 0/C. Treatments performed on day 1 after culture were Cs-137 irradiation (10, 20, or 40 Gy); ADR (10, 30, or 100 micrograms/ml); DHAQ (5, 20, or 50 micrograms/ml); or various combinations of drugs and radiation. Hearts were checked every day for functional activity as evidenced by continuous heart best. Untreated hearts beat rhythmically for up to 9 days (average = 6.8 days); treated hearts stopped beating between 2 and 7 days after treatment. Using this endpoint of functional retention time (FRT), dose response curves were obtained for all individual agents. Combinations of ADR and DHAQ (at concentrations that resulted in FRTs of 3.5 days) produced no greater effect than either agent alone. However, the combination of radiation (FRT = 5.3 days) with ADR, DHAQ or both drugs was more effective than was drug alone. This system may help to predict the cardiotoxic effects that result from the use of these drugs and radiation.

  11. 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

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

    PubMed

    Contreras, Deisy; Arumugaswami, Vaithilingaraja

    2016-08-23

    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.

  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. 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.

  15. 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.

  16. 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

  17. Gene expression in mouse ovarian follicle development in vivo versus an ex vivo alginate culture system.

    PubMed

    Parrish, Elizabeth M; Siletz, Anaar; Xu, Min; Woodruff, Teresa K; Shea, Lonnie D

    2011-08-01

    Ovarian follicle maturation results from a complex interplay of endocrine, paracrine, and direct cell-cell interactions. This study compared the dynamic expression of key developmental genes during folliculogenesis in vivo and during in vitro culture in a 3D alginate hydrogel system. Candidate gene expression profiles were measured within mouse two-layered secondary follicles, multi-layered secondary follicles, and cumulus-oocyte complexes (COCs). The expression of 20 genes involved in endocrine communication, growth signaling, and oocyte development was investigated by real-time PCR. Gene product levels were compared between i) follicles of similar stage and ii) COCs derived either in vivo or by in vitro culture. For follicles cultured for 4 days, the expression pattern and the expression level of 12 genes were the same in vivo and in vitro. Some endocrine (cytochrome P450, family 19, subfamily A, polypeptide 1 (Cyp19a1) and inhibin βA subunit (Inhba)) and growth-related genes (bone morphogenetic protein 15 (Bmp15), kit ligand (Kitl), and transforming growth factor β receptor 2 (Tgfbr2)) were downregulated relative to in vivo follicles. For COCs obtained from cultured follicles, endocrine-related genes (inhibin α-subunit (Inha) and Inhba) had increased expression relative to in vivo counterparts, whereas growth-related genes (Bmp15, growth differentiation factor 9, and kit oncogene (Kit)) and zona pellucida genes were decreased. However, most of the oocyte-specific genes (e.g. factor in the germline α (Figla), jagged 1 (Jag1), and Nlrp5 (Mater)) were expressed in vitro at the same level and with the same pattern as in vivo-derived follicles. These studies establish the similarities and differences between in vivo and in vitro cultured follicles, guiding the creation of environments that maximize follicle development and oocyte quality. PMID:21610168

  18. 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

  19. 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.

  20. 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

  1. 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.

  2. 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.

  3. 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

  4. 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.

  5. 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

  6. 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.

  7. 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.

  8. A novel anaerobic co-culture system for bio-hydrogen production from sugarcane bagasse.

    PubMed

    Cheng, Jingrong; Zhu, Mingjun

    2013-09-01

    A novel co-culture of Clostridium thermocellum and Thermoanaerobacterium aotearoense with pretreated sugarcane bagasse (SCB) under mild alkali conditions for bio-hydrogen production was established, exhibiting a cost-effective and synergetic advantage in bio-hydrogen production over monoculture of C. thermocellum or T. aotearoense with untreated SCB. The optimized pretreatment conditions were established to be 3% NaOH, and a liquid to solid ratio of 25:1 at 80°C for 3h. A final hydrogen production of 50.05±1.51 mmol/L was achieved with 40 g/L pretreated SCB at 55°C. The established co-culture system provides a novel consolidated bio-processing strategy for bioconversion of SCB to bio-hydrogen. PMID:23899575

  9. 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).

  10. 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

  11. A novel anaerobic co-culture system for bio-hydrogen production from sugarcane bagasse.

    PubMed

    Cheng, Jingrong; Zhu, Mingjun

    2013-09-01

    A novel co-culture of Clostridium thermocellum and Thermoanaerobacterium aotearoense with pretreated sugarcane bagasse (SCB) under mild alkali conditions for bio-hydrogen production was established, exhibiting a cost-effective and synergetic advantage in bio-hydrogen production over monoculture of C. thermocellum or T. aotearoense with untreated SCB. The optimized pretreatment conditions were established to be 3% NaOH, and a liquid to solid ratio of 25:1 at 80°C for 3h. A final hydrogen production of 50.05±1.51 mmol/L was achieved with 40 g/L pretreated SCB at 55°C. The established co-culture system provides a novel consolidated bio-processing strategy for bioconversion of SCB to bio-hydrogen.

  12. 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

  13. 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…

  14. Early Childhood Education in Iceland: A Response to the Welfare System, the Natural Environment, the Cultural Heritage and Foreign Influences.

    ERIC Educational Resources Information Center

    Einarsdottir, Johanna

    This paper discusses early childhood education in Iceland and its in relation to the country's cultural heritage, the family welfare system, and legislation and influences from abroad. Because Iceland has been isolated for centuries from the rest of the world, the country has developed its own culture and maintained its native language. The need…

  15. 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…

  16. The famous versus the inconvenient - or the dawn and the rise of 3D-culture systems

    PubMed Central

    Altmann, Brigitte; Welle, Alexander; Giselbrecht, Stefan; Truckenmüller, Roman; Gottwald, Eric

    2009-01-01

    One of the greatest impacts on in vitro cell biology was the introduction of three-dimensional (3D) culture systems more than six decades ago and this era may be called the dawn of 3D-tissue culture. Although the advantages were obvious, this field of research was a “sleeping beauty” until the 1970s when multicellular spheroids were discovered as ideal tumor models. With this rebirth, organotypical culture systems became valuable tools and this trend continues to increase. While in the beginning, simple approaches, such as aggregation culture techniques, were favored due to their simplicity and convenience, now more sophisticated systems are used and are still being developed. One of the boosts in the development of new culture techniques arises from elaborate manufacturing and surface modification techniques, especially micro and nano system technologies that have either improved dramatically or have evolved very recently. With the help of these tools, it will soon be possible to generate even more sophisticated and more organotypic-like culture systems. Since 3D perfused or superfused systems are much more complex to set up and maintain compared to use of petri dishes and culture flasks, the added value of 3D approaches still needs to be demonstrated. PMID:21607106

  17. 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

  18. The potential of a polyphasic PCR-dGGE approach in evaluating microbial diversity of natural whey cultures for water-buffalo Mozzarella cheese production: bias of culture-dependent and culture-independent analyses.

    PubMed

    Ercolini, D; Moschetti, G; Blaiotta, G; Coppola, S

    2001-12-01

    A polyphasic PCR-DGGE approach was used to describe the microbial population occurring in natural whey cultures (NWCs) for water-buffalo Mozzarella cheese production. Total microbial community was assessed without cultivation by analyzing DNA directly extracted from the original samples of NWC. In addition, DNA extracted from bulks of cells formed by harvesting colonies from the serial dilution agar plates of a variety of culture media was used to profile the "cultivable" community. The 16S rDNA V3 region was amplified using DNA from NWC as well as DNA from bulks as templates and the amplicons were separated by DGGE. The microbial entities occurring in NWCs were identified by partial 16S rDNA sequencing of DGGE bands: four lactic acid bacteria (LAB) closest relative of Streptococcus thermophilus, Lactococcus lactis, Lactobacillus delbrueckii and Lactobacillus crispatus were revealed by the analysis of DNA directly extracted from NWC while two other LAB, Lactobacillus fermentum and Enterococcus faecalis, were identified by analyzing DNA from the cultivable community. The developed PCR-DGGE analysis of the "cultivable" community showed good potential in evaluating microbial diversity of a dairy environment: it usefully highlighted the bias introduced by selective amplification when compared to the analysis of the total community from NWC and allowed suitability of media and growth conditions to be evaluated. Moreover, it could be used to complete the culture independent study of microbial diversity to give information on concentration ratios among species occurring in a particular environment and can be proposed for rapid identification of dominant microorganisms in alternative to traditional tools.

  19. 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

  20. 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)

  1. 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

  2. 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

  3. Diabetic embryopathy: studies using a rat embryo culture system and an animal model.

    PubMed

    Akazawa, Shoichi

    2005-09-01

    The mechanism of diabetic embryopathy was investigated using in vitro experiments in a rat embryo culture system and in streptozotocin-induced diabetic pregnant rats. The energy metabolism in embryos during early organogenesis was characterized by a high rate of glucose utilization and lactic acid production (anaerobic glycolysis). Embryos uninterruptedly underwent glycolysis. When embryos were cultured with hypoglycemic serum, such embryos showed malformations in association with a significant reduction in glycolysis. In a diabetic environment, hyperglycemia caused an increased glucose flux into embryonic cells without a down-regulation of GLUT1 and an increased metabolic overload on mitochondria, leading to an increased formation of reactive oxygen species (ROS). Activation of the hexamine pathway, subsequently occurring with increased protein carbonylation and increased lipid peroxidation, also contributed to the increased generation of ROS. Hyperglycemia also caused a myo-inositol deficiency with a competitive inhibition of ambient glucose, which might have been associated with a diminished phosphoinositide signal transduction. In the presence of low activity of the mitochondrial oxidative glucose metabolism, the ROS scavenging system in the embryo was not sufficiently developed. Diabetes further weakened the antioxidant system, especially, the enzyme for GSH synthesis, gamma-GCS, thereby reducing the GSH concentration. GSH depletion also disturbed prostaglandin biosynthesis. An increased formation of ROS in a diminished GSH-dependent antioxidant system may, therefore, play an important role in the development of embryonic malformations in diabetes.

  4. 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.

  5. Quorum-sensing regulation of constitutive plantaricin by Lactobacillus plantarum strains under a model system for vegetables and fruits.

    PubMed

    Rizzello, Carlo G; Filannino, Pasquale; Di Cagno, Raffaella; 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.

  6. 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

  7. 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

  8. 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

  9. 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.

  10. Adipose tissue engineering in three-dimensional levitation tissue culture system based on magnetic nanoparticles.

    PubMed

    Daquinag, Alexes C; Souza, Glauco R; Kolonin, Mikhail G

    2013-05-01

    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

  11. Adipose tissue engineering in three-dimensional levitation tissue culture system based on magnetic nanoparticles.

    PubMed

    Daquinag, Alexes C; Souza, Glauco R; Kolonin, Mikhail G

    2013-05-01

    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

  12. Evaluation of a rat tracheal epithelial cell culture assay system to identify respiratory carcinogens

    SciTech Connect

    Steele, V.E.; Arnold, J.T.; Arnold, J.V.; Mass, M.J. )

    1989-01-01

    To evaluate a short-term epithelial cell assay system to detect respiratory carcinogens, primary cultures of rat tracheal epithelial cells were exposed to a series of 17 compounds and scored for morphologically transformed cell colonies 28 days later. The test compounds included known carcinogens and noncarcinogens in volatile or liquid form. Tracheal epithelial cells were isolate from F344 rats, plated onto collagen-coated dishes, and exposed to the test compounds on day 1 for 24 hours. At day 30 the cultures were fixed, stained, and scored for colonies having a density greater than 1,300 cells/mm{sup 2}. With standardized protocols, such colonies are very infrequent in media and solvent control cultures. Concentration levels for each chemical were chosen over a range from nontoxic to toxic levels. Highly positive compounds in this assay included benzo(a)pyrene, benzo(l)aceanthrylene, 3-methylcholanthrene, and formaldehyde. Compounds which were negative in this assay included pyrene, benzo(e)pyrene, and 4-nitroquinoline-N-oxide. Examining the concordance of in vitro results with whole animal carcinogenesis studies revealed an accuracy of 88% with one false-positive and one false-negative compound. The results of these studies indicate that the rat tracheal epithelial cell assay may be useful in identifying potential respiratory carcinogens in our environment.

  13. 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

  14. 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

  15. Ex vivo organ culture of human hair follicles: a model epithelial-neuroectodermal-mesenchymal interaction system.

    PubMed

    Tobin, Desmond J

    2011-01-01

    The development of hair follicle organ culture techniques is a significant milestone in cutaneous biology research. The hair follicle, or more accurately the "pilo-sebaceous unit", encapsulates all the important physiologic processes found in the human body; controlled cell growth/death, interactions between cells of different histologic type, cell differentiation and migration, and hormone responsitivity to name a few. Thus, the value of the hair follicle as a model for biological scientific research goes way beyond its scope for cutaneous biology or dermatology alone. Indeed, the recent and dramatic upturn in interest in hair follicle biology has focused principally on the pursuit of two of biology's holy grails; post-embryonic morphogenesis and control of cyclical tissue activity. The hair follicle organ culture model, pioneered by Philpott and colleagues, ushered in an exceptionally accessible way to assess how cells of epithelial (e.g., keratinocytes), mesenchymal (e.g., fibroblasts), and neuroectodermal (e.g., melanocytes) origin interact in a three-dimensional manner. Moreover, this assay system allows us to assess how various natural and pharmacologic agents affect complex tissues for growth modulation. In this article, I focus on the culture of the human hair follicle mini-organ, discussing both the practical issues involved and some possible research applications of this assay.

  16. Microfluidic co-culture system for cancer migratory analysis and anti-metastatic drugs screening

    PubMed Central

    Mi, Shengli; Du, Zhichang; Xu, Yuanyuan; Wu, Zhengjie; Qian, Xiang; Zhang, Min; Sun, Wei

    2016-01-01

    Tumour metastasis is an important reason for cancer death, and cancer cell migration is an important step in the process of tumour metastasis. Studying cancer cell migration is of great significance. Here, we present a novel microfluidic co-culture system and establish mild, moderate and severe cancer models by using HMEpiC and MDA-MB–231 cells to study cancer cell migration and anti-cancer drug screening. Using this device, we achieved high cell viability (over 90%) and a stable analysis of the migration ability of cancer cells. We observed that the density of the cancer cells determined the probability of the occurrence of metastatic cells and that the induction of normal cells affected the metastatic velocity of each cancer cell. We verified that the increase in the migration ability of MDA-MB-231 cells co-cultured with HMEpiC cells was relative to the increased secretion of IL-6 and that this was verified by an IL-6 inhibitor assay. This co-culture also led to decreased CK-14 secretion and morphological changes in HMEpiC cells. Finally, significant inhibition of paclitaxel and tamoxifen on cancer migration was observed. Taken together, our microfluidic device could be a useful tool for the quantitation of the migratory capability and anti-metastatic drug screening. PMID:27762336

  17. Hepatocyte function within a stacked double sandwich culture plate cylindrical bioreactor for bioartificial liver system.

    PubMed

    Xia, Lei; Arooz, Talha; Zhang, Shufang; Tuo, Xiaoye; Xiao, Guangfa; Susanto, Thomas Adi Kurnia; Sundararajan, Janani; Cheng, Tianming; Kang, Yuzhan; Poh, Hee Joo; Leo, Hwa Liang; Yu, Hanry

    2012-11-01

    Bioartificial liver (BAL) system is promising as an alternative treatment for liver failure. We have developed a bioreactor with stacked sandwich culture plates for the application of BAL. This bioreactor design addresses some of the persistent problems in flat-bed bioreactors through increasing cell packing capacity, eliminating dead flow, regulating shear stress, and facilitating the scalability of the bioreactor unit. The bioreactor contained a stack of twelve double-sandwich-culture plates, allowing 100 million hepatocytes to be housed in a single cylindrical bioreactor unit (7 cm of height and 5.5 cm of inner diameter). The serial flow perfusion through the bioreactor increased cell-fluid contact area for effective mass exchange. With the optimal perfusion flow rate, shear stress was minimized to achieve high and uniform cell viabilities across different plates in the bioreactor. Our results demonstrated that hepatocytes cultured in the bioreactor could re-establish cell polarity and maintain liver-specific functions (e.g. albumin and urea synthesis, phase I&II metabolism functions) for seven days. The single bioreactor unit can be readily scaled up to house adequate number of functional hepatocytes for BAL development.

  18. 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

  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 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.

  1. 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

  2. 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

  3. 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

  4. 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

  5. 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.

  6. 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

  7. Multilineage differentiation of rhesus monkey embryonic stem cells in three-dimensional culture systems.

    PubMed

    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. PMID:12743323

  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. 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

  10. 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

    2014-05-23

    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.

  11. 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

  12. 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

  13. A microfluidic cell culture system for monitoring of sequential changes in endothelial cells after heat stress.

    PubMed

    Tazawa, Hidekatsu; Sato, Kenjiro; Tsutiya, Atsuhiro; Tokeshi, Manabu; Ohtani-Kaneko, Ritsuko

    2015-08-01

    Endothelial damage induced by a highly elevated body temperature is crucial in some diseases including viral hemorrhagic fevers. Here, we report the heat-induced sequential changes of endothelial cells under shear stress, which were determined with a microfluidic culture system. Although live cell imaging showed only minor changes in the appearance of heat-treated cells, Hsp70 mRNA expression analysis demonstrated that the endothelial cells in channels of the system responded well to heat treatment. F-actin staining also revealed clear changes in the bundles of actin filaments after heat treatment. Well-organized bundles of actin filaments in control cells disappeared in heat-treated cells cultured in the channel. Furthermore, the system enabled detection of sequential changes in plasminogen activator inhibitor-1 (PAI-1) secretion from endothelial cells. PAI-1 concentration in the effluent solution was significantly elevated for the first 15min after initiation of heat treatment, and then decreased subsequently. This study provides fundamental information on heat-induced endothelial changes under shear stress and introduces a potent tool for analyzing endothelial secretions. PMID:26044666

  14. Biomass recycle as a means to improve the energy efficiency of CELSS algal culture systems

    NASA Astrophysics Data System (ADS)

    Radmer, R.; Cox, J.; Lieberman, D.; Behrens, P.; Arnett, K.

    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. We report experiments in which cultures of the algae 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 we tested hydrolyzed waste biomass from these same algae 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.

  15. Evaluation of Biocompatibility of Alloplastic Materials: Development of a Tissue Culture In Vitro Test System.

    PubMed

    Gerullis, Holger; Georgas, Evangelos; Eimer, Christoph; Goretzki, Peter E; Lammers, Bernhard J; Klosterhalfen, Bernd; Boros, Mihaly; Wishahi, Mohamed; Heusch, Gerd; Otto, Thomas

    2011-12-01

    Optimized biocompatibility is a major requirement for alloplastic materials currently applied in surgical approaches for hernia, incontinence, and prolapse situations. Tissue ingrowth/adherence and formation of connective tissue seem to have important influence in mesh incorporation at the implant site. In an in vitro approach we randomly investigated 7 different mesh types currently used in surgeries with various indications with regard to their adherence performance. Using a tissue culture approach, meshes were incubated with tissue representative of fibroblasts, muscle cells, and endothelial cells originating from 10 different patients. After 6 weeks, the meshes were assessed microscopically and a ranking of their adherence performance was established. Tissue culture was successful in 100% of the probes. We did not remark on interindividual differences concerning the growth and adherence performance after incubation with the different meshes in the investigated 10 patients. The ranking was consistent in all patients. In this test system, PVDF Dynamesh® (FEG Textiltechnik, Aachen, Germany) was the mesh with the best growth-in score. The test system was feasible and reproducible. Pore size seems to be a predictor of adherence performance. The test system may be a helpful tool for further investigations, and the predictive value should be assessed in further in vitro and in vivo experiments. PMID:22504966

  16. A microfluidic cell culture system for monitoring of sequential changes in endothelial cells after heat stress.

    PubMed

    Tazawa, Hidekatsu; Sato, Kenjiro; Tsutiya, Atsuhiro; Tokeshi, Manabu; Ohtani-Kaneko, Ritsuko

    2015-08-01

    Endothelial damage induced by a highly elevated body temperature is crucial in some diseases including viral hemorrhagic fevers. Here, we report the heat-induced sequential changes of endothelial cells under shear stress, which were determined with a microfluidic culture system. Although live cell imaging showed only minor changes in the appearance of heat-treated cells, Hsp70 mRNA expression analysis demonstrated that the endothelial cells in channels of the system responded well to heat treatment. F-actin staining also revealed clear changes in the bundles of actin filaments after heat treatment. Well-organized bundles of actin filaments in control cells disappeared in heat-treated cells cultured in the channel. Furthermore, the system enabled detection of sequential changes in plasminogen activator inhibitor-1 (PAI-1) secretion from endothelial cells. PAI-1 concentration in the effluent solution was significantly elevated for the first 15min after initiation of heat treatment, and then decreased subsequently. This study provides fundamental information on heat-induced endothelial changes under shear stress and introduces a potent tool for analyzing endothelial secretions.

  17. Biomass recycle as a means to improve the energy efficiency of CELSS algal culture systems.

    PubMed

    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. We report experiments in which cultures of the algae 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 we tested hydrolyzed waste biomass from these same algae 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.

  18. Framing a socio-indexical basis for the emergence and cultural transmission of phonological systems

    PubMed Central

    Plummer, Andrew R.; Beckman, Mary E.

    2015-01-01

    Moulin-Frier et al. (2016) proffer a conceptual framework and computational modeling architecture for the investigation of the emergence of phonological universals for spoken languages. They validate the framework and architecture by testing to see whether universals such as the prevalence of triangular vowel systems that show adequate dispersion in the F1-F2-F3 space can fall out of simulations of referential communication between social agents, without building principles such as dispersion directly into the model. In this paper, we examine the assumptions underlying the framework, beginning with the assumption that it is such substantive universals that are in need of explanation rather than the rich diversity of phonological systems observed across human cultures and the compositional (“prosodic”) structure that characterizes signed as well as spoken languages. Also, when emergence is construed at the time-scales of the biological evolution of the species and of the cultural evolution of distinct speech communities, it is the affiliative or affective rather than the referential function that has the greater significance for our understanding of how phonological systems can emerge de novo in ontogeny. PMID:26834297

  19. 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

  20. 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.

  1. 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

  2. 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.

  3. 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

  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. Usability, Communicability and Cultural Tourism in Interactive Systems: Trends, Economic Effects and Social Impact

    NASA Astrophysics Data System (ADS)

    Cipolla Ficarra, Francisco V.; Nicol, Emma; Cipolla-Ficarra, Miguel

    We have developed a set of techniques and an analysis methodology aimed at boosting the quality of interactive tourism systems. The details of it will be presented in full and with real examples which have yielded interesting results in the last few years, both from the social and economical point of view, but with a huge wealth of cultural and natural heritage. We will also present a first guidelines to foster tourism in those villages that are willing to promote themselves in the national and international market at a low cost.

  7. Testing a theory of organizational culture, climate and youth outcomes in child welfare systems: a United States national study.

    PubMed

    Williams, Nathaniel J; Glisson, Charles

    2014-04-01

    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

  8. 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

  9. 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.

  10. 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

  11. A 3D Sphere Culture System Containing Functional Polymers for Large-Scale Human Pluripotent Stem Cell Production

    PubMed Central

    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

    2014-01-01

    Summary 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

  12. 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

  13. 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

  14. An improved system for exposure of cultured mammalian cells to gaseous compounds in the chromosomal aberration assay.

    PubMed

    Asakura, Masumi; Sasaki, Toshiaki; Sugiyama, Toshie; Arito, Heihachiro; Fukushima, Shoji; Matsushima, Taijiro

    2008-04-30

    A gas exposure system using rotating vessels was improved for exposure of cultured mammalian cells to gaseous compounds in the chromosomal aberration assay. This system was composed of 12 square culture vessels, a device for preparation of air containing test gas, and positive and negative control gases at target concentrations and for supplying these gases to the culture vessels, and a roller apparatus in an incubator. Chinese hamster lung cells (CHL/IU) were grown on one side of the inner surface of the square culture vessel in the MEM medium. Immediately prior to exposure, the medium was changed to the modified MEM. Air in the culture vessel was replaced with air containing test gas, positive or negative control gas. Then, the culture vessels were rotated at 1.0 rpm. The monolayered culture cells were exposed to test gas during about 3/4 rotation at upper positions and alternatively immersed into the culture medium during about 1/4 rotation at lower positions. This system allowed the chromosomal aberration assay simultaneously at least at three different concentrations of a test gas together with positive and negative control gases with and without metabolic activations, and duplicate culture at each exposure concentration. Seven gaseous compounds, 1,3-butadiene, chlorodifluoromethane, ethyl chloride, methyl bromide, methyl chloride, propyne, and vinyl chloride, none of which has been tested to date, were tested on CHL/IU for the chromosomal aberration assay using this gas exposure system. All the compounds except chlorodifluoromethane showed positive responses of the structural chromosomal aberrations, whereas polyploidy was not induced by any of these gases. This improved gas exposure system proved to be useful for detecting chromosomal aberrations of gaseous compounds.

  15. An improved system for exposure of cultured mammalian cells to gaseous compounds in the chromosomal aberration assay.

    PubMed

    Asakura, Masumi; Sasaki, Toshiaki; Sugiyama, Toshie; Arito, Heihachiro; Fukushima, Shoji; Matsushima, Taijiro

    2008-04-30

    A gas exposure system using rotating vessels was improved for exposure of cultured mammalian cells to gaseous compounds in the chromosomal aberration assay. This system was composed of 12 square culture vessels, a device for preparation of air containing test gas, and positive and negative control gases at target concentrations and for supplying these gases to the culture vessels, and a roller apparatus in an incubator. Chinese hamster lung cells (CHL/IU) were grown on one side of the inner surface of the square culture vessel in the MEM medium. Immediately prior to exposure, the medium was changed to the modified MEM. Air in the culture vessel was replaced with air containing test gas, positive or negative control gas. Then, the culture vessels were rotated at 1.0 rpm. The monolayered culture cells were exposed to test gas during about 3/4 rotation at upper positions and alternatively immersed into the culture medium during about 1/4 rotation at lower positions. This system allowed the chromosomal aberration assay simultaneously at least at three different concentrations of a test gas together with positive and negative control gases with and without metabolic activations, and duplicate culture at each exposure concentration. Seven gaseous compounds, 1,3-butadiene, chlorodifluoromethane, ethyl chloride, methyl bromide, methyl chloride, propyne, and vinyl chloride, none of which has been tested to date, were tested on CHL/IU for the chromosomal aberration assay using this gas exposure system. All the compounds except chlorodifluoromethane showed positive responses of the structural chromosomal aberrations, whereas polyploidy was not induced by any of these gases. This improved gas exposure system proved to be useful for detecting chromosomal aberrations of gaseous compounds. PMID:18342567

  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. Prevention of the degeneration of human dopaminergic neurons in an astrocyte co-culture system allowing endogenous drug metabolism

    PubMed Central

    Efremova, Liudmila; Schildknecht, Stefan; Adam, Martina; Pape, Regina; Gutbier, Simon; Hanf, Benjamin; Bürkle, Alexander; Leist, Marcel

    2015-01-01

    Background and Purpose Few neuropharmacological model systems use human neurons. Moreover, available test systems rarely reflect functional roles of co-cultured glial cells. There is no human in vitro counterpart of the widely used 1-methyl-4-phenyl-tetrahydropyridine (MPTP) mouse model of Parkinson's disease Experimental Approach We generated such a model by growing an intricate network of human dopaminergic neurons on a dense layer of astrocytes. In these co-cultures, MPTP was metabolized to 1-methyl-4-phenyl-pyridinium (MPP+) by the glial cells, and the toxic metabolite was taken up through the dopamine transporter into neurons. Cell viability was measured biochemically and by quantitative neurite imaging, siRNA techniques were also used. Key Results We initially characterized the activation of PARP. As in mouse models, MPTP exposure induced (poly-ADP-ribose) synthesis and neurodegeneration was blocked by PARP inhibitors. Several different putative neuroprotectants were then compared in mono-cultures and co-cultures. Rho kinase inhibitors worked in both models; CEP1347, ascorbic acid or a caspase inhibitor protected mono-cultures from MPP+ toxicity, but did not protect co-cultures, when used alone or in combination. Application of GSSG prevented degeneration in co-cultures, but not in mono-cultures. The surprisingly different pharmacological profiles of the models suggest that the presence of glial cells, and the in situ generation of the toxic metabolite MPP+ within the layered cultures played an important role in neuroprotection. Conclusions and Implications Our new model system is a closer model of human brain tissue than conventional cultures. Its use for screening of candidate neuroprotectants may increase the predictiveness of a test battery. PMID:25989025

  19. Evaluation of FilmArray and Verigene Systems for Rapid Identification of Positive Blood Cultures

    PubMed Central

    Bhatti, M. M.; Boonlayangoor, S.; Beavis, K. G.

    2014-01-01

    The Verigene tests for Gram-positive and Gram-negative organisms in blood culture and the FilmArray blood culture identification panel were assessed for their ability to identify pathogens from positive blood cultures. Both platforms correctly identified bacteria in 92% of monomicrobial cultures analyzed, with times to identification that were significantly shorter than those for identification from subcultures. PMID:25031445

  20. Fabrication of micropatterned hydrogels for neural culture systems using dynamic mask projection photolithography.

    PubMed

    Curley, J Lowry; Jennings, Scott R; Moore, Michael J

    2011-01-01

    photocrosslinkable or self polymerizing hydrogel, or selectively adhered to the permeable support membrane using cell-restrictive photopatterning. Using the DMD, we created hydrogel constructs up to ~1mm thick, but thin film (<200 μm) PEG structures were limited by oxygen quenching of the free radical polymerization reaction. We subsequently developed a technique utilizing a layer of oil above the polymerization liquid which allowed thin PEG structure polymerization. In this protocol, we describe the expeditious creation of 3D hydrogel systems for production of microfabricated neural cell and tissue cultures. The dual hydrogel constructs demonstrated herein represent versatile in vitro models that may prove useful for studies in neuroscience involving cell survival, migration, and/or neurite growth and guidance. Moreover, as the protocol can work for many types of hydrogels and cells, the potential applications are both varied and vast.

  1. Critical Incident Stress Management (CISM) in complex systems: cultural adaptation and safety impacts in healthcare.

    PubMed

    Müller-Leonhardt, Alice; Mitchell, Shannon G; Vogt, Joachim; Schürmann, Tim

    2014-07-01

    In complex systems, such as hospitals or air traffic control operations, critical incidents (CIs) are unavoidable. These incidents can not only become critical for victims but also for professionals working at the "sharp end" who may have to deal with critical incident stress (CIS) reactions that may be severe and impede emotional, physical, cognitive and social functioning. These CIS reactions may occur not only under exceptional conditions but also during every-day work and become an important safety issue. In contrast to air traffic management (ATM) operations in Europe, which have readily adopted critical incident stress management (CISM), most hospitals have not yet implemented comprehensive peer support programs. This survey was conducted in 2010 at the only European general hospital setting which implemented CISM program since 2004. The aim of the article is to describe possible contribution of CISM in hospital settings framed from the perspective of organizational safety and individual health for healthcare professionals. Findings affirm that daily work related incidents also can become critical for healthcare professionals. Program efficiency appears to be influenced by the professional culture, as well as organizational structure and policies. Overall, findings demonstrate that the adaptation of the CISM program in general hospitals takes time but, once established, it may serve as a mechanism for changing professional culture, thereby permitting the framing of even small incidents or near misses as an opportunity to provide valuable feedback to the system.

  2. 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

  3. Studies of rumen function in an in vitro continuous culture system.

    PubMed

    Merry, R J; Smith, R H; McAllan, A B

    1987-06-01

    An in vitro continuous culture system to simulate processes in the rumen is described. This comprises a culture vessel (Fig. 2) fed continuously with pelleted solid feed (Fig. 1); artificial saliva also enters the vessel while effluent leaves it continuously in two streams, one is filtered and the other simply overflows (Fig. 1). In this way liquid and solid turnover times may be manipulated independently; in a trial experiment the former and latter were 6.3% and 3.4% per hour respectively. In four replicated experiments a steady-state was achieved in 5-6 days with a feed of barley, tapioca, fishmeal, urea and straw and maintained for a further 7 to 14 days. During a steady-state period, rumen characteristics in terms of protozoal count, pH, redox potential, total volatile fatty acid concentration and ammonia concentration were stable and similar to those found in the rumens of animals given similar diets. Flows of microbial N compounds were assessed by infused Na2H32PO4 with the artificial saliva and estimating the 32P label incorporated in bacterial fractions. The system demonstrably provided a satisfactory means for studying many aspects of rumen function.

  4. Application of three-dimensional culture systems to study mammalian spermatogenesis, with an emphasis on the rhesus monkey (Macaca mulatta)

    PubMed Central

    Huleihel, Mahmoud; Nourashrafeddin, Seyedmehdi; Plant, Tony M

    2015-01-01

    In vitro culture of spermatogonial stem cells (SSCs) has generally been performed using two-dimensional (2D) culture systems; however, such cultures have not led to the development of complete spermatogenesis. It seems that 2D systems do not replicate optimal conditions of the seminiferous tubules (including those generated by the SSC niche) and necessary for spermatogenesis. Recently, one of our laboratories has been able to induce proliferation and differentiation of mouse testicular germ cells to meiotic and postmeiotic stages including generation of sperm in a 3D soft agar culture system (SACS) and a 3D methylcellulose culture system (MCS). It was suggested that SACS and MCS form a special 3D microenvironment that mimics germ cell niche formation in the seminiferous tubules, and thus permits mouse spermatogenesis in vitro. In this review, we (1) provide a brief overview of the differences in spermatogenesis in rodents and primates, (2) summarize data related to attempts to generate sperm in vitro, (3) report for the first time formation of colonies/clusters of cells and differentiation of meiotic (expression of CREM-1) and postmeiotic (expression of acrosin) germ cells from undifferentiated spermatogonia isolated from the testis of prepubertal rhesus monkeys and cultured in SACS and MCS, and (4) indicate research needed to optimize 3D systems for in vitro primate spermatogenesis and for possible future application to man. PMID:26067870

  5. 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

  6. 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

  7. 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

  8. Enzyme-based flow injection analysis system for glutamine and glutamate in mammalian cell culture media.

    PubMed

    Mayer, C; Frauer, A; Schalkhammer, T; Pittner, F

    1999-03-01

    We present the setup of a flow injection analysis system designed for on-line monitoring of glutamate and glutamine. These amino acids represent a major energy source in mammalian cell culture. A cycling assay consisting of glutamate dehydrogenase and aspartate aminotransferase produces NADH proportional to the glutamate concentration in the sample. NADH is then measured spectrophotometrically. Glutamine is determined by conversion to glutamate which is fed into the cycling assay. The conversion of glutamine to glutamate is catalyzed by asparaginase. Asparaginase was used in place of glutaminase due to its relatively high reactivity with glutamine and a pH optimum similar to that of glutamate dehydrogenase. The enzymes were immobilized covalently to activated controlled pore glass beads and integrated into the flow injection analysis system. The application of the immobilized enzymes and the technical setup are presented in this paper.

  9. The effect of sevoflurane on developing A/J strain mouse embryos using a whole-embryo culture system--the incidence of cleft lip in culture embryos.

    PubMed

    Yamada, Morimasa; Yamamoto, Naoki; Ohgami, Saori; Kanazawa, Mayuko; Harada, Jun; Ohno, Norikazu; Natsume, Nagato

    2014-03-01

    A/J strain mice have a high spontaneous incidence of cleft lip (ICL) and/or palate. The primary palate-related effects of sevoflurane on developing A/J strain mouse embryos (embryos) were studied using a whole-embryo culture (WEC) system. This system could separate the direct effects of sevoflurane from those that are maternally mediated. A total of 205 10.5-d embryos were cultured for 24 h in either a control group (control gas: 95% O2 and 5% CO2) or sevoflurane-administered groups (1/4, 1/2, and 1 minimum alveolar concentration (MAC) with control gas) for 8 h. After 16 h, 11.5-d culture embryos were examined in terms of crown-rump length, number of somites, and protein content. Crown-rump length in the 1 MAC was significantly shorter than in the control group (p < 0.05). Protein content in the 1/2 MAC (p < 0.05) and 1 MAC (p < 0.001) was significantly lower than in the control group. The ICL showed no significant differences between each group. (The ICL rose with an increase in the sevoflurane concentration, but this was not significant). The positive findings in this study indicate that a WEC system is useful for studying the mechanisms of ICL (teratogenicity) associated with sevoflurane.

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

    PubMed

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

    2012-06-01

    Services to families have traditionally been delivered in a medical model. This presents challenges including workforce shortages, lack of cultural diversity, lack of training in strength-based work, and difficulty in successfully engaging and retaining families in the therapy process. The system of care (SOC) effort has worked to establish formal roles for caregivers in SOC to improve services. This paper provides an example of one community's efforts to change the SOC by expanding the roles available to caregivers in creating systems change. It describes the model developed by Communities of Care (CoC), a SOC in Central Massachusetts, and its evolution over a 10 year period. First person accounts by system partners, caregivers hired into professional roles as well as a family receiving services, demonstrate how hiring caregivers at all levels can change systems and change lives, not only for those being served but for the caregiver/professionals doing the work. It also demonstrates, however, that change at the system level is incremental, takes time, and can be fleeting unless an ongoing effort is made to support and sustain those changes. PMID:21947874

  11. 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.

  12. 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.

  13. 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.

  14. 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

  15. Endothelin-1 release from cultured endothelial cells induced by sera from patients with systemic lupus erythematosus.

    PubMed Central

    Yoshio, T; Masuyama, J; Mimori, A; Takeda, A; Minota, S; Kano, S

    1995-01-01

    OBJECTIVES--To clarify the pathophysiological role of endothelin-1 (ET-1) in the vascular injury associated with systemic lupus erythematosus (SLE) by investigating the effect of sera from patients with SLE on ET-1 release from cultured human umbilical vein endothelial cells. METHODS--Confluent monolayers of cultured human umbilical vein endothelial cells were incubated with serum samples (diluted 1:10) from 25 patients with SLE and 16 normal controls for two hours at 37 degrees C and ET-1 concentration in the culture supernatant was measured by enzyme immunoassay. RESULTS--The mean release of ET-1 from endothelial cells in the presence of serum from SLE patients was greater than in the presence of serum from normal controls (p < 0.005). ET-1 release from endothelial cells significantly correlated with the titre of IgM anti-endothelial cell antibodies (IgM-AECA) and immune complex concentration in sera from SLE patients (p < 0.05 and p < 0.01, respectively). After gel chromatography of the serum from an SLE patient, those fractions containing IgM-AECA or immune complex were shown to stimulate ET-1 release from endothelial cells. Heat aggregated IgG also stimulated ET-1 release from endothelial cells in a concentration dependent manner. CONCLUSIONS--IgM-AECA and immune complexes may stimulate ET-1 release from endothelial cells and ET-1 may play an important role in the initiation and development of vascular injury, such as pulmonary hypertension and lupus nephritis, in SLE. PMID:7794041

  16. Regulation of the transferrin-independent iron transport system in cultured cells.

    PubMed

    Kaplan, J; Jordan, I; Sturrock, A

    1991-02-15

    Mammalian cells accumulate iron via the binding of transferrin to high affinity surface receptors, or through a transferrin-independent pathway which involves the uptake of iron-organic anion chelates by a membrane-based transport system. Previously we determined that the transferrin-independent transport system was present on a wide variety of cultured cells (Sturrock, A., Alexander, J., Lamb, J., Craven, C. M., and Kaplan, J. (1990) J. Biol. Chem. 265, 3139-3145). In this communication we demonstrate that the transferrin-independent iron uptake system is regulated differently than the transferrin-mediated pathway. The activity of the transferrin-independent system was unaffected by changes in cellular growth rate, induction of DNA synthesis and cell division, or depletion of cellular iron. Exposure of cells to ferric or ferrous iron, however, resulted in a time-dependent increase in transport activity, due to a change in Vmax with no change in Km. Increased transport activity was seen in a variety of cultured cell types, occurred in the presence of cycloheximide, and persisted for hours after removal of iron. The ability of other transition metals to induce changes in transport, or to compete with iron for accumulation by the transferrin-independent uptake system, was critically dependent on the composition of the media in which the cells were incubated. Metals such as Cu2+ or Zn2+, but not Cd2+ or Mn2+, when dissolved in a balanced salt solution buffered with 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, induced changes in the transferrin-independent iron transport system. The same metals which induced changes in transport were ineffective in media containing amino acids, ascorbate, or N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]glycine. The Vmax of the transferrin-independent iron transport system was also elevated by increases in intracellular Ca2+. The effect of iron on transport activity, however, did not result from an iron-induced release of

  17. An axenic plant culture system for optimal growth in long-term studies.

    PubMed

    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. CDII) 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 a 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 microg d(-1). Fumaric, malic, malonic, oxalic, and succinic acids were measured as components of the root exudates.

  18. 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.

  19. 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.

  20. 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

  1. 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

    2012-04-02

    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. 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

  3. 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.

  4. 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

  5. A high-throughput drop microfluidic system for virus culture and analysis.

    PubMed

    Fischer, Audrey E; Wu, Susan K; Proescher, Jody B G; Rotem, Assaf; Chang, Connie B; Zhang, Huidan; Tao, Ye; Mehoke, Thomas S; Thielen, Peter M; Kolawole, Abimbola O; Smith, Thomas J; Wobus, Christiane E; Weitz, David A; Lin, Jeffrey S; Feldman, Andrew B; Wolfe, Joshua T

    2015-03-01

    High mutation rates and short replication times lead to rapid evolution in RNA viruses. New tools for high-throughput culture and analysis of viral phenotypes will enable more effective studies of viral evolutionary processes. A water-in-oil drop microfluidic system to study virus-cell interactions at the single event level on a massively parallel scale is described here. Murine norovirus (MNV-1) particles were co-encapsulated with individual RAW 264.7 cells in 65 pL aqueous drops formed by flow focusing in 50 μm microchannels. At low multiplicity of infection (MOI), viral titers increased greatly, reaching a maximum 18 h post-encapsulation. This system was employed to evaluate MNV-1 escape from a neutralizing monoclonal antibody (clone A6.2). Further, the system was validated as a means for testing escape from antibody neutralization using a series of viral point mutants. Finally, the replicative capacity of single viral particles in drops under antibody stress was tested. Under standard conditions, many RNA virus stocks harbor minority populations of genotypic and phenotypic variants, resulting in quasispecies. These data show that when single cells are encapsulated with single viral particles under antibody stress without competition from other virions, the number of resulting infectious particles is nearly equivalent to the number of viral genomes present. These findings suggest that lower fitness virions can infect cells successfully and replicate, indicating that the microfluidics system may serve as an effective tool for isolating mutants that escape evolutionary stressors.

  6. 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

  7. 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

  8. 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…

  9. A Phenotypic Culture System for the Molecular Analysis of CNS Myelination in the Spinal Cord

    PubMed Central

    Davis, Hedvika; Gonzalez, Mercedes; Stancescu, Maria; Love, Rachal; Hickman, James J.; Lambert, Stephen

    2014-01-01

    Studies of central nervous system myelination lack defined in vitro models which would effectively dissect molecular mechanisms of myelination that contain cells of the correct phenotype. Here we describe a co-culture of purified motoneurons and oligodendrocyte progenitor cells, isolated from rat embryonic spinal cord using a combination of immunopanning techniques. This model illustrates differentiation of oligodendrocyte progenitors into fully functional mature oligodendrocytes that myelinate axons. It also illustrates a contribution of axons to the rate of oligodendrocyte maturation and myelin gene expression. The defined conditions used allow molecular analysis of distinct stages of myelination and precise manipulation of inductive cues affecting axonal–oligodendrocyte interactions. This phenotypic in vitro myelination model can provide valuable insight into our understanding of demyelinating disorders, such as multiple sclerosis and traumatic diseases such as spinal cord injury where demyelination represents a contributing factor to the pathology of the disorder. PMID:25064806

  10. 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.

  11. Sensitivity analysis and parameter identification of nonlinear hybrid systems for glycerol transport mechanisms in continuous culture.

    PubMed

    Gao, Kuikui; Zhang, Xu; Feng, Enmin; Xiu, Zhilong

    2014-04-21

    In this paper, we establish a modified fourteen-dimensional nonlinear hybrid dynamic system with genetic regulation to describe the microbial continuous culture, in which we consider that there are three possible ways for glycerol to pass the cell's membrane and one way for 1,3-PD (passive diffusion and active transport). Then we discuss the existence, uniqueness, continuous dependence of solutions and the compactness of the solution set. We construct a global sensitivity analysis approach to reduce the number of kinetic parameters. In order to infer the most reasonable transport mechanism of glycerol, we propose a parameter identification model aiming at identifying the parameter with higher sensitivity and transport of glycerol, which takes the robustness index of the intracellular substance together with the relative error between the experimental data and the computational values of the extracellular substance as a performance index. Finally, a parallel algorithm is applied to find the optimal transport of glycerol and parameters. PMID:24406809

  12. A phenotypic culture system for the molecular analysis of CNS myelination in the spinal cord.

    PubMed

    Davis, Hedvika; Gonzalez, Mercedes; Stancescu, Maria; Love, Rachal; Hickman, James J; Lambert, Stephen

    2014-10-01

    Studies of central nervous system myelination lack defined in vitro models which would effectively dissect molecular mechanisms of myelination that contain cells of the correct phenotype. Here we describe a co-culture of purified motoneurons and oligodendrocyte progenitor cells, isolated from rat embryonic spinal cord using a combination of immunopanning techniques. This model illustrates differentiation of oligodendrocyte progenitors into fully functional mature oligodendrocytes that myelinate axons. It also illustrates a contribution of axons to the rate of oligodendrocyte maturation and myelin gene expression. The defined conditions used allow molecular analysis of distinct stages of myelination and precise manipulation of inductive cues affecting axonal-oligodendrocyte interactions. This phenotypic in vitro myelination model can provide valuable insight into our understanding of demyelinating disorders, such as multiple sclerosis and traumatic diseases such as spinal cord injury where demyelination represents a contributing factor to the pathology of the disorder.

  13. The tracing of mycobacteria in drinking water supply systems by culture, conventional, and real time PCRs.

    PubMed

    Klanicova, Barbora; Seda, Jaromir; Slana, Iva; Slany, Michal; Pavlik, Ivo

    2013-12-01

    Mycobacteria are widely present in diverse aquatic habitats, where they can survive for months or years while some species can even proliferate. The resistance of different mycobacterial species to disinfection methods like chlorination or ozonation could result in their presence in the final tap water of consumers. In this study, the culture method, Mycobacterium tuberculosis complex conventional duplex PCR for detection of non-tuberculous mycobacteria (NTM) and quantitative real-time PCR (qPCR) to detect three subspecies of M. avium species (M. a. avium, M. a. hominissuis, and M. a. paratuberculosis) were used to trace their possible path of transmission from the watershed through the reservoir and drinking water plant to raw drinking water and finally to households. A total of 124 samples from four drinking water supply systems in the Czech Republic, 52 dam sediments, 34 water treatment plant sludge samples, and 38 tap water household sediments, were analyzed. NTM of 11 different species were isolated by culture from 42 (33.9 %) samples; the most prevalent were M. gordonae (16.7 %), M. triplex (14.3 %), M. lentiflavum (9.5 %), M. a. avium (7.1 %), M. montefiorenase (7.1 %), and M. nonchromogenicum (7.1 %). NTM DNA was detected in 92 (76.7 %) samples. By qPCR analysis a statistically significant decrease (P < 0.01) was observed along the route from the reservoir (dam sediments), through water treatment sludge and finally to household sediments. The concentrations ranged from 10(0) to 10(4) DNA cells/g. It was confirmed that drinking water supply systems (watershed-reservoir-drinking water treatment plant-household) might be a potential transmission route for mycobacteria.

  14. A hydroponic rice seedling culture model system for investigating proteome of salt stress in rice leaf.

    PubMed

    Kim, Dea-Wook; Rakwal, Randeep; Agrawal, Ganesh Kumar; Jung, Young-Ho; Shibato, Junko; Jwa, Nam-Soo; Iwahashi, Yumiko; Iwahashi, Hitoshi; Kim, Du Hyun; Shim, Ie-Sung; Usui, Kenji

    2005-12-01

    By using an in vivo hydroponic rice seedling culture system, we investigated the physiological and biochemical responses of a model rice japonica cultivar Nipponbare to salt stress using proteomics and classical biochemical methods. Yoshida's nutrient solution (YS) was used to grow rice seedlings. YS-grown 18-day-old seedlings manifested highly stable and reproducible symptoms, prominently the wilting and browning of the 3rd leaf, reduced photosynthetic activity, inhibition in overall seedling growth, and failure to develop new (5th) leaf, when subjected to salt stress by transferring them to YS containing 130 mM NaCl for 4 days. As leaf response to salt stress is least investigated in rice by proteomics, we used the 3rd leaf as source material. A comparison of 2-DE protein profiles between the untreated control and salt-stressed 3rd leaves revealed 55 differentially expressed CBB-stained spots, where 47 spots were increased over the control. Of these changed spots, the identity of 33 protein spots (27 increased and 5 decreased) was determined by nESI-LC-MS/MS. Most of these identified proteins belonged to major metabolic processes like photosynthetic carbon dioxide assimilation and photorespiration, suggesting a good correlation between salt stress-responsive proteins and leaf morphology. Moreover, 2-DE immunoblot and enzymatic activity analyses of 3rd leaves revealed remarkable changes in the key marker enzymes associated with oxidative damage to salt stress: ascorbate peroxidase and lipid peroxidation were induced, and catalase was suppressed. These results demonstrate that hydroponic culture system is best suited for proteomics of salt stress in rice seedling.

  15. 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

  16. 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

  17. Development and use of culture systems to modulate specific cell responses

    NASA Astrophysics Data System (ADS)

    Martin, Yves

    Culture surfaces that induce specific localized cell responses are required to achieve tissue-like cell growth in three-dimensional (3D) environments, as well as to develop more efficient cell-based diagnostic techniques, noticeably when working with fragile cells such as stem cells or platelets. As such, Chapter 1 of this thesis work is devoted to the review of 3D cell-material interactions in vitro and the corresponding existing culture systems available to achieve in vivo-like cell responses. More adequate 3D culture systems will need to be developed to mimic several characteristics of in vivo environments, including lowered non-specific cell-material interactions and localized biochemical signaling. The experimental work in this thesis is based on the hypothesis that well-studied and optimized surface treatments will be able to lower non-specific cell-material interactions and allow local chemical modification in order to achieve specific localized cell-material interactions for different applications. As such, in Chapter 2 and Chapter 3 of this thesis, surface treatments were developed using plasma polymerization and covalent immobilization of a low-fouling polymer (i.e., poly(ethylene glycol)) and characterized and optimized using a large number of techniques including atomic force microscopy, quartz crystal microbalance, surface plasmon resonance, x-ray photoelectron spectroscopy and fluorescence-based techniques. The main plasma polymerization parameter important for surface chemical content, specifically nitrogen to carbon content, was identified as being glow discharge power, while reaction time and power determined plasma film thickness. Moreover, plasma films were shown to be stable in aqueous environments. Covalently-bound poly(ethylene glycol) (PEG) layers physicochemical and mechanical properties are dependent on fabrication methods. Polymer concentration in solution is an important indicator of final layer properties, and use of a theta solvent

  18. Use of the whole-embryo culture system in drug safety assessment?

    PubMed

    Bechter, R; Schön, H

    1988-01-01

    The embryotoxicity and, in particular, the teratogenic potential of three new drug combinations were investigated. The interactions of the single components (all already on the market) in the different drug mixtures were determined by testing the single components in addition to the drug combinations. In order to evaluate the usefulness of these results for safety assessment, the no-effect concentrations (NEC) of the single compounds or the mixed drug preparations, with respect to embryonic growth and differentiation and the presence of anomalies, were compared with human plasma/serum levels during therapeutic use. In addition, in vivo animal data of the single components were used to put into perspective the NEC values after direct in vitro exposure of embryos. Embryotoxicity, measured as a retardation of growth and of differentiation, as well as an increased percentage of cultured embryos with morphological abnormalities, was present at concentrations of 100 mug/ml for the preparation KT 1/300 (ketotifen/theophylline). These effects could be attributed to the component theophylline, as ketotifen was free of an embryotoxic/teratogenic potential in vitro. In the case of the compound mix VKB/BQ, a combination of pindolol, clopamide and endralazine, the in vitro effects of the mixture on embryonic development could also be attributed to one of the compounds (pindolol) in the mixture. In the case of HYCT 4520, the effects of the components hydergine (co-dergocrine mesylate), clopamide and triamterene on embryonic growth, differentiation and anomalies were additive in the drug mixture. None of the compounds was reported to be associated with human teratogenicity. The NEC of the components in the whole-embryo culture system, tested either as a mixture or as individual compounds, were higher than the human peak plasma levels measured during therapeutic use of the compounds. In addition, the single components were reported to be non-teratogenic in in vivo teratogenicity

  19. Probing functional properties of nociceptive axons using a microfluidic culture system.

    PubMed

    Tsantoulas, Christoforos; Farmer, Clare; Machado, Patricia; Baba, Katsuhiro; McMahon, Stephen B; Raouf, Ramin

    2013-01-01

    Pathological changes in axonal function are integral features of many neurological disorders, yet our knowledge of the molecular basis of axonal dysfunction remains limited. Microfluidic chambers (MFCs) can provide unique insight into the axonal compartment independent of the soma. Here we demonstrate how an MFC based cell culture system can be readily adapted for the study of axonal function in vitro. We illustrate the ease and versatility to assay electrogenesis and conduction of action potentials (APs) in naïve, damaged or sensitized DRG axons using calcium imaging at the soma for pharmacological screening or patch-clamp electrophysiology for detailed biophysical characterisation. To demonstrate the adaptability of the system, we report by way of example functional changes in nociceptor axons following sensitization by neurotrophins and axotomy in vitro. We show that NGF can locally sensitize axonal responses to capsaicin, independent of the soma. Axotomizing neurons in MFC results in a significant increase in the proportion of neurons that respond to axonal stimulation, and interestingly leads to accumulation of Nav1.8 channels in regenerating axons. Axotomy also augmented AP amplitude following axotomy and altered activation thresholds in a subpopulation of regenerating axons. We further show how the system can readily be used to study modulation of axonal function by non-neuronal cells such as keratinocytes. Hence we describe a novel in vitro platform for the study of axonal function and a surrogate model for nerve injury and sensitization. PMID:24278311

  20. 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.

  1. Closed system cell culture protocol using HYPERStack vessels with gas permeable material technology.

    PubMed

    Titus, Kim; Klimovich, Vitaly; Rothenberg, Mark; Pardo, Pilar; Tanner, Allison; Martin, Greg

    2010-11-29

    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.

  2. 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

  3. 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.

  4. 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.

  5. 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.

  6. 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

  7. 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.

  8. 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

  9. Simple suspension culture system of human iPS cells maintaining their pluripotency for cardiac cell sheet engineering.

    PubMed

    Haraguchi, Yuji; Matsuura, Katsuhisa; Shimizu, Tatsuya; Yamato, Masayuki; Okano, Teruo

    2015-12-01

    In this study, a simple three-dimensional (3D) suspension culture method for the expansion and cardiac differentiation of human induced pluripotent stem cells (hiPSCs) is reported. The culture methods were easily adapted from two-dimensional (2D) to 3D culture without any additional manipulations. When hiPSCs were directly applied to 3D culture from 2D in a single-cell suspension, only a few aggregated cells were observed. However, after 3 days, culture of the small hiPSC aggregates in a spinner flask at the optimal agitation rate created aggregates which were capable of cell passages from the single-cell suspension. Cell numbers increased to approximately 10-fold after 12 days of culture. The undifferentiated state of expanded hiPSCs was confirmed by flow cytometry, immunocytochemistry and quantitative RT-PCR, and the hiPSCs differentiated into three germ layers. When the hiPSCs were subsequently cultured in a flask using cardiac differentiation medium, expression of cardiac cell-specific genes and beating cardiomyocytes were observed. Furthermore, the culture of hiPSCs on Matrigel-coated dishes with serum-free medium containing activin A, BMP4 and FGF-2 enabled it to generate robust spontaneous beating cardiomyocytes and these cells expressed several cardiac cell-related genes, including HCN4, MLC-2a and MLC-2v. This suggests that the expanded hiPSCs might maintain the potential to differentiate into several types of cardiomyocytes, including pacemakers. Moreover, when cardiac cell sheets were fabricated using differentiated cardiomyocytes, they beat spontaneously and synchronously, indicating electrically communicative tissue. This simple culture system might enable the generation of sufficient amounts of beating cardiomyocytes for use in cardiac regenerative medicine and tissue engineering.

  10. Axenic culture of a candidate division TM7 bacterium from the human oral cavity and biofilm interactions with other oral bacteria.

    PubMed

    Soro, Valeria; Dutton, Lindsay C; Sprague, Susan V; Nobbs, Angela H; Ireland, Anthony J; Sandy, Jonathan R; Jepson, Mark A; Micaroni, Massimo; Splatt, Peter R; Dymock, David; Jenkinson, Howard F

    2014-10-01

    The diversity of bacterial species in the human oral cavity is well recognized, but a high proportion of them are presently uncultivable. Candidate division TM7 bacteria are almost always detected in metagenomic studies but have not yet been cultivated. In this paper, we identified candidate division TM7 bacterial phylotypes in mature plaque samples from around orthodontic bonds in subjects undergoing orthodontic treatment. Successive rounds of enrichment in laboratory media led to the isolation of a pure culture of one of these candidate division TM7 phylotypes. The bacteria formed filaments of 20 to 200 μm in length within agar plate colonies and in monospecies biofilms on salivary pellicle and exhibited some unusual morphological characteristics by transmission electron microscopy, including a trilaminated cell surface layer and dense cytoplasmic deposits. Proteomic analyses of cell wall protein extracts identified abundant polypeptides predicted from the TM7 partial genomic sequence. Pleiomorphic phenotypes were observed when the candidate division TM7 bacterium was grown in dual-species biofilms with representatives of six different oral bacterial genera. The TM7 bacterium formed long filaments in dual-species biofilm communities with Actinomyces oris or Fusobacterium nucleatum. However, the TM7 isolate grew as short rods or cocci in dual-species biofilms with Porphyromonas gingivalis, Prevotella intermedia, Parvimonas micra, or Streptococcus gordonii, forming notably robust biofilms with the latter two species. The ability to cultivate TM7 axenically should majorly advance understanding of the physiology, genetics, and virulence properties of this novel candidate division oral bacterium.

  11. Bioavailability and ecotoxicity of arsenic species in solution culture and soil system: implications to remediation.

    PubMed

    Bolan, Nanthi; Mahimairaja, Santiago; Kunhikrishnan, Anitha; Seshadri, Balaji; Thangarajan, Ramya

    2015-06-01

    In this work, bioavailability and ecotoxicity of arsenite (As(III)) and arsenate (As(V)) species were compared between solution culture and soil system. Firstly, the adsorption of As(III) and As(V) was compared using a number of non-allophanic and allophanic soils. Secondly, the bioavailability and ecotoxicity were examined using germination, phytoavailability, earthworm, and soil microbial activity tests. Both As-spiked soils and As-contaminated sheep dip soils were used to test bioavailability and ecotoxicity. The sheep dip soil which contained predominantly As(V) species was subject to flooding to reduce As(V) to As(III) and then used along with the control treatment soil to compare the bioavailability between As species. Adsorption of As(V) was much higher than that of As(III), and the difference in adsorption between these two species was more pronounced in the allophanic than non-allophanic soils. In the solution culture, there was no significant difference in bioavailability and ecotoxicity, as measured by germination and phytoavailability tests, between these two As species. Whereas in the As-spiked soils, the bioavailability and ecotoxicity were higher for As(III) than As(V), and the difference was more pronounced in the allophanic than non-allophanic soils. Bioavailability of As increased with the flooding of the sheep dip soils which may be attributed to the reduction of As(V) to As(III) species. The results in this study have demonstrated that while in solution, the bioavailability and ecotoxicity do not vary between As(III) and As(V), in soils, the latter species is less bioavailable than the former species because As(V) is more strongly retained than As(III). Since the bioavailability and ecotoxicity of As depend on the nature of As species present in the environment, risk-based remediation approach should aim at controlling the dynamics of As transformation.

  12. 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.

  13. 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.

  14. 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