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Sample records for culture independent methods

  1. Culture-independent methods for identifying microbial communities in cheese

    OpenAIRE

    Jany, Jean-Luc; Barbier, Georges

    2008-01-01

    International audience; This review focuses on the culture-independent methods available for the description of both bacterial and fungal communities in cheese. Important steps of the culture-independent strategy, which relies on bulk DNA extraction from cheese and polymerase chain reaction (PCR) amplification of selected sequences, are discussed. We critically evaluate the identification techniques already used for monitoring microbial communities in cheese, including PCR-denaturing gradient...

  2. Culture-dependent and culture-independent methods reveal diverse methylotrophic communities in terrestrial environments

    OpenAIRE

    Eyice, Özge; Schäfer, Hendrik

    2016-01-01

    One-carbon compounds such as methanol, dimethylsulfide (DMS) and dimethylsulfoxide (DMSO) are significant intermediates in biogeochemical cycles. They are suggested to affect atmospheric chemistry and global climate. Methylotrophic microorganisms are considered as a significant sink for these compounds; therefore, we analyzed the diversity of terrestrial bacteria that utilize methanol, DMS and DMSO as carbon and energy source using culture-dependent and culture-independent methods. The effect...

  3. Microbial composition during Chinese soy sauce koji-making based on culture dependent and independent methods.

    Science.gov (United States)

    Yan, Yin-zhuo; Qian, Yu-lin; Ji, Feng-di; Chen, Jing-yu; Han, Bei-zhong

    2013-05-01

    Koji-making is a key process for production of high quality soy sauce. The microbial composition during koji-making was investigated by culture-dependent and culture-independent methods to determine predominant bacterial and fungal populations. The culture-dependent methods used were direct culture and colony morphology observation, and PCR amplification of 16S/26S rDNA fragments followed by sequencing analysis. The culture-independent method was based on the analysis of 16S/26S rDNA clone libraries. There were differences between the results obtained by different methods. However, sufficient overlap existed between the different methods to identify potentially significant microbial groups. 16 and 20 different bacterial species were identified using culture-dependent and culture-independent methods, respectively. 7 species could be identified by both methods. The most predominant bacterial genera were Weissella and Staphylococcus. Both 6 different fungal species were identified using culture-dependent and culture-independent methods, respectively. Only 3 species could be identified by both sets of methods. The most predominant fungi were Aspergillus and Candida species. This work illustrated the importance of a comprehensive polyphasic approach in the analysis of microbial composition during soy sauce koji-making, the knowledge of which will enable further optimization of microbial composition and quality control of koji to upgrade Chinese traditional soy sauce product.

  4. Diversity of endophytic bacteria of Dendrobium officinale based on culture-dependent and culture-independent methods

    Directory of Open Access Journals (Sweden)

    Cong Pei

    2017-01-01

    Full Text Available Culture-dependent and culture-independent methods were compared and evaluated in the study of the endophytic diversity of Dendrobium officinale. Culture-independent methods consisted of polymerase chain reaction–denaturing gradient gel electrophoresis (PCR-DGGE and metagenome methods. According to the results, differences were found between the three methods. Three phyla, namely Firmicutes, Proteobacteria, and Actinobacteria, were detected using the culture-dependent method, and two phyla, Firmicutes and Proteobacteria, were detected by the DGGE method. Using the metagenome method, four major phyla were determined, including Proteobacteria (76.54%, Actinobacteria (18.56%, Firmicutes (2.27%, and Bacteroidetes (1.56%. A distinct trend was obtained at the genus level in terms of the method and the corresponding number of genera determined. There were 449 genera and 16 genera obtained from the metagenome and DGGE methods, respectively, and only 7 genera were obtained through the culture-dependent method. By comparison, all the genera from the culture-dependent and DGGE methods were contained in the members determined using the metagenome method. Overall, culture-dependent methods are limited to ‘finding’ endophytic bacteria in plants. DGGE is an alternative to investigating primary diversity patterns; however, the metagenome method is still the best choice for determining the endophytic profile in plants. It is essential to use multiphasic approaches to study cultured and uncultured microbes.

  5. Characterization of microbes in prosthetic joint specimens by culture-independent molecular methods

    DEFF Research Database (Denmark)

    Xu, Yijuan; Rudkjøbing, Vibeke Børsholt; Simonsen, Ole

    was to investigate the microbial diversity in surgical samples (eg. synovial fluid, periprosthetic tissue, removed prosthesis) from 22 prosthetic patients using a range of culture-independent molecular methods including broad range 16S rRNA gene PCR, cloning, phylogeny, quantitative PCR (qPCR), and fluorescence...... in situ hybridization (FISH). Concomitant samples were cultured by standard methods. Overall, the results of culture-based and molecular methods showed concordant results for 13 patients and discrepant results for 6 patients. In the remaining cases, culture methods identified one species or a group...... approach, and FISH and confocal scanning laser microscopy visualized the presence of both single cells and microcolonies. In conclusion, cultureindependent methods identified more species and more polymicrobial infections than standard culturebased methods. Little is presently known about the pathogenesis...

  6. Comparison of culture-dependent and -independent methods for bacterial community monitoring during Montasio cheese manufacturing.

    Science.gov (United States)

    Carraro, Lisa; Maifreni, Michela; Bartolomeoli, Ingrid; Martino, Maria Elena; Novelli, Enrico; Frigo, Francesca; Marino, Marilena; Cardazzo, Barbara

    2011-04-01

    The microbial community in milk is of great importance in the manufacture of traditional cheeses produced using raw milk and natural cultures. During milk curdling and cheese ripening, complex interactions occur in the microbial community, and accurate identification of the microorganisms involved provides essential information for understanding their role in these processes and in flavor production. Recent improvements in molecular biological methods have led to their application to food matrices, and thereby opened new perspectives for the study of microbial communities in fermented foods. In this study, a description of microbial community composition during the manufacture and ripening of Montasio cheese was provided. A combined approach using culture-dependent and -independent methods was applied. Culture-dependent identification was compared with 16S clone libraries sequencing data obtained from both DNA and reverse-transcribed RNA (cDNA) amplification and real-time quantitative PCR (qPCR) assays developed to detect and quantify specific bacterial species/genera (Streptococcus thermophilus, Lactobacillus casei, Pediococcus pentosaceus, Enterococcus spp., Pseudomonas spp.). S. thermophilus was the predominant LAB species throughout the entire ripening period of Montasio cheese. The culture-independent method demonstrates the relevant presence of Pseudomonas spp. and Lactococcus piscium at the beginning of ripening. The culture-dependent approach and the two culture-independent approaches produced complementary information, together generating a general view of cheese microbial ecology.

  7. Towards a culturally independent participatory design method: Fusing game elements into the design process

    DEFF Research Database (Denmark)

    Jensen, Mika Yasuoka; Nakatani, Momoko; Ohno, Takehiko

    2013-01-01

    Historically, Participatory Design (PD) was introduced and applied in the Scandinavian and American context as a practical design method for collective creativity and stakeholder involvement. In this paper, by fusing game elements into PD, we suggest a first step towards a culturally independent PD...... method called the ICT Service Design Game to ease the prevailing concern that PD has limited applicability in other cultural settings. We conduct four experiments on ICT Service Design Game in Scandinavia and Asia to evaluate its feasibility. The experiments identify some differences in the PD process...... and the created Persona characteristics, probably due to cultural differences. Of more interestingly, despite these process differences, the game elements of our PD variant were found to promote the key values of PD in culturally diverse settings by stimulating idea creation and participation. Our experiments...

  8. Towards a culturally independent participatory design method: Fusing game elements into the design process

    DEFF Research Database (Denmark)

    Jensen, Mika Yasuoka; Nakatani, Momoko; Ohno, Takehiko

    2013-01-01

    method called the ICT Service Design Game to ease the prevailing concern that PD has limited applicability in other cultural settings. We conduct four experiments on ICT Service Design Game in Scandinavia and Asia to evaluate its feasibility. The experiments identify some differences in the PD process......Historically, Participatory Design (PD) was introduced and applied in the Scandinavian and American context as a practical design method for collective creativity and stakeholder involvement. In this paper, by fusing game elements into PD, we suggest a first step towards a culturally independent PD...... and the created Persona characteristics, probably due to cultural differences. Of more interestingly, despite these process differences, the game elements of our PD variant were found to promote the key values of PD in culturally diverse settings by stimulating idea creation and participation. Our experiments...

  9. Culture-Dependent and -Independent Methods Capture Different Microbial Community Fractions in Hydrocarbon-Contaminated Soils

    Science.gov (United States)

    Stefani, Franck O. P.; Bell, Terrence H.; Marchand, Charlotte; de la Providencia, Ivan E.; El Yassimi, Abdel; St-Arnaud, Marc; Hijri, Mohamed

    2015-01-01

    Bioremediation is a cost-effective and sustainable approach for treating polluted soils, but our ability to improve on current bioremediation strategies depends on our ability to isolate microorganisms from these soils. Although culturing is widely used in bioremediation research and applications, it is unknown whether the composition of cultured isolates closely mirrors the indigenous microbial community from contaminated soils. To assess this, we paired culture-independent (454-pyrosequencing of total soil DNA) with culture-dependent (isolation using seven different growth media) techniques to analyse the bacterial and fungal communities from hydrocarbon-contaminated soils. Although bacterial and fungal rarefaction curves were saturated for both methods, only 2.4% and 8.2% of the bacterial and fungal OTUs, respectively, were shared between datasets. Isolated taxa increased the total recovered species richness by only 2% for bacteria and 5% for fungi. Interestingly, none of the bacteria that we isolated were representative of the major bacterial OTUs recovered by 454-pyrosequencing. Isolation of fungi was moderately more effective at capturing the dominant OTUs observed by culture-independent analysis, as 3 of 31 cultured fungal strains ranked among the 20 most abundant fungal OTUs in the 454-pyrosequencing dataset. This study is one of the most comprehensive comparisons of microbial communities from hydrocarbon-contaminated soils using both isolation and high-throughput sequencing methods. PMID:26053848

  10. Culture-Dependent and -Independent Methods Capture Different Microbial Community Fractions in Hydrocarbon-Contaminated Soils.

    Directory of Open Access Journals (Sweden)

    Franck O P Stefani

    Full Text Available Bioremediation is a cost-effective and sustainable approach for treating polluted soils, but our ability to improve on current bioremediation strategies depends on our ability to isolate microorganisms from these soils. Although culturing is widely used in bioremediation research and applications, it is unknown whether the composition of cultured isolates closely mirrors the indigenous microbial community from contaminated soils. To assess this, we paired culture-independent (454-pyrosequencing of total soil DNA with culture-dependent (isolation using seven different growth media techniques to analyse the bacterial and fungal communities from hydrocarbon-contaminated soils. Although bacterial and fungal rarefaction curves were saturated for both methods, only 2.4% and 8.2% of the bacterial and fungal OTUs, respectively, were shared between datasets. Isolated taxa increased the total recovered species richness by only 2% for bacteria and 5% for fungi. Interestingly, none of the bacteria that we isolated were representative of the major bacterial OTUs recovered by 454-pyrosequencing. Isolation of fungi was moderately more effective at capturing the dominant OTUs observed by culture-independent analysis, as 3 of 31 cultured fungal strains ranked among the 20 most abundant fungal OTUs in the 454-pyrosequencing dataset. This study is one of the most comprehensive comparisons of microbial communities from hydrocarbon-contaminated soils using both isolation and high-throughput sequencing methods.

  11. Combination of culture-dependent and culture-independent molecular methods for the determination of lactic microbiota in sucuk.

    Science.gov (United States)

    Kesmen, Z; Yetiman, A E; Gulluce, A; Kacmaz, N; Sagdic, O; Cetin, B; Adiguzel, A; Sahin, F; Yetim, H

    2012-02-15

    In this study, the culture-dependent and culture-independent molecular methods were used for the identification of lactic acid bacteria (LAB) in sucuk a Turkish fermented dry sausage. On the one hand, the PCR-DGGE method targetting the V1 and V3 regions of 16S DNA was applied to DNA that was directly extracted from sucuk samples. On the other hand, rep-PCR fingerprinting was performed for the primary differentiation and grouping of the isolates, and the results were confirmed by sequencing of the 16S rDNA and 16S-23S rDNA intergenic spacer region. As a result of the PCR-DGGE analysis of all the samples, total 8 different lactic acid bacteria were identified, and Lactobacillus sakei, Lactobacillus curvatus and Weissella viridescens were the dominant microbiota among these bacteria. The culture-dependent approach indicated that the majority of the strains belonged to the Lactobacillus genera including Lb. sakei, Lactobacillus plantarum, Lb. curvatus, Lactobacillus brevis, Lactobacillus farciminis and Lactobacillus alimentarius. However, Leuconostoc and Weisella were also detected as minor genera. Again, Lactococcus piscium, Weissella halotolerans, Staphylococcus succinus and the comigrated Staphylococcus piscifermentans/Staphylococcus condimenti/Staphylococcus carnosus group were detected only with the culture-independent method while Lb. plantarum, Leuconostoc mesenteroides and Leuconostoc citreum were identified only by using the culture-dependent method. In the results, it was concluded that the combination of culture-dependent and culture-independent methods was necessary for reliable and detailed investigation of LAB communities in fermented food products.

  12. Determination of lactic microflora of kefir grains and kefir beverage by using culture-dependent and culture-independent methods.

    Science.gov (United States)

    Kesmen, Zülal; Kacmaz, Nazife

    2011-01-01

    In this study, we investigated the bacterial compositions of kefir grains and kefir beverages collected from different regions of Turkey by using culture-independent and culture-dependent methods. In the culture-independent detection, 10 different species of bacteria were detected in total by using the polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis of the 16S rRNA gene V3 region. Among these species, Lactobacillus kefiranofaciens was the most dominant one in the kefir grains, while Lactococcus lactis was found to be significantly prevalent in the kefir beverages. In the culture-dependent detection, the primary differentiation and grouping of the isolates from kefir beverages and kefir grains were performed using repetitive sequence-based PCR (rep-PCR) fingerprinting, and the results were validated by 16S rDNA full-length sequencing. According to the results of culture-dependent methods, the most frequently isolated species were L. lactis, Leuconostoc mesenteroides, and Lactobacillus kefiri, respectively. Only 3 species, which are L. lactis, Lactobacillus acidophilus, and Streptococcus thermophilus, were detected with both culture-dependent and culture-independent methods. This study showed that the combination of both methods is necessary for a detailed and reliable investigation of microbial communities in kefir grains and kefir beverages. Due to their artisan- and region-dependent microflora, kefir products can be a source of interesting lactic acid bacteria, either new taxa or strains with specific functional properties, which might be used for the development of new starter cultures and innovative food products. Therefore, an increasing demand exists for new strains that show desirable effects on the product characteristics Artisan dairy products are a candidate source of such microorganisms. For this reason, in this study, the bacterial compositions of kefir grains and kefir beverages obtained from

  13. A culture independent method for the detection of Aeromonas sp. from water samples

    Directory of Open Access Journals (Sweden)

    Fadua Latif-Eugenín

    2016-01-01

    Full Text Available The genus Aeromonas is present in a wide variety of water environments and is recognised as potentially pathogenic to humans and animals. Members of this genus are often confused with Vibrio when using automated, commercial identification systems that are culture-dependent. This study describes a polymerase chain reaction (PCR detection method for Aeromonas that is culture- independent and that targets the glycerophospholopid-cholesterol acyltransferase (gcat gene, which is specific for this genus. The GCAT-PCR was 100% specific in artificially inoculated water samples, with a detection limit that ranged from 2.5 to 25 cfu/mL. The success at detecting this pathogen in 86 water samples using the GCAT-PCR method was identical to the conventional culturing method when a pre-enrichment step was carried out, yielding 83.7% positive samples. On the other hand, without a pre-enrichment step, only 77.9% of the samples were positive by culturing and only 15.1% with the GCATPCR. However, 83.7% positive samples were obtained for the GCAT-PCR when the water volume for the DNA extraction was increased from 400 μL to 4 mL. The proposed molecular method is much faster (5 or 29 h than the culturing method (24 or 48 h whether performed directly or after a pre-enrichment step and it will enable the fast detection of Aeromonas in water samples helping to prevent a possible transmission to humans.

  14. Bacterial diversity determination using culture-dependent and culture-independent methods

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    M. Ghiasian

    2017-04-01

    Full Text Available Mud volcanoes are taken into consideration by geologists and oil industry experts have given their association with oil and gas reserves and methane greenhouse gas production in hydrosphere and atmosphere. Gomishan mud volcano phenomenon in the southeastern edge of the Caspian Sea, given its oil and gas resources, has been studied by some geologists in terms of geology and tectonics but not in terms of microbiology. Accordingly, it seems necessary to study this phenomenon from the perspective of microbiology in order to identify prokaryotes living in this area. Prokaryotes diversity in Mud volcano has been studied by cultivation techniques, fluorescence in situ hybridization, and denaturing gradient gel electrophoresis of PCR-amplified fragments of 16S rRNA genes. Total cell abundance in the mud volcano from 1×101-6×101per milliliter was determined by 4', 6-diamidino-2-phenylindole direct count. The detectable proportion of Archaea to Bacteria in the community by FISH was one to five. High viable counts (1 – 3 × 106 were obtained in culture media. A total of 122 isolates were obtained, 46 colonies were selected based on primarily morphological and physiological traits, and their 16S rRNA sequences were determined. The isolated genera included Halomonas (20%, Arthrobacter (5%, Kocuria (5%, Thalassobacillus (5%, Marinobacter (20%, Paracoccus (5%, Roseovarius (5%, Jeotgalicoccus (5%, Bacillus (15%, and Staphylococcus (15%. Regarding DGGE analysis, selected bands were obtained from the gels, reamplified and sequenced. Overall, 75% of the bacterial sequences were related to Rahnella and 25% related to Serratia.

  15. Diversity of Termitomyces associated with fungus-farming termites assessed by cultural and culture-independent methods.

    Science.gov (United States)

    Makonde, Huxley M; Boga, Hamadi I; Osiemo, Zipporah; Mwirichia, Romano; Stielow, J Benjamin; Göker, Markus; Klenk, Hans-Peter

    2013-01-01

    Fungus-cultivating termites make use of an obligate mutualism with fungi from the genus Termitomyces, which are acquired through either vertical transmission via reproductive alates or horizontally transmitted during the formation of new mounds. Termitomyces taxonomy, and thus estimating diversity and host specificity of these fungi, is challenging because fruiting bodies are rarely found. Molecular techniques can be applied but need not necessarily yield the same outcome than morphological identification. Culture-dependent and culture-independent methods were used to comprehensively assess host specificity and gut fungal diversity. Termites were identified using mitochondrial cytochrome oxidase II (COII) genes. Twenty-three Termitomyces cultures were isolated from fungal combs. Internal transcribed spacer (ITS) clone libraries were constructed from termite guts. Presence of Termitomyces was confirmed using specific and universal primers. Termitomyces species boundaries were estimated by cross-comparison of macromorphological and sequence features, and ITS clustering parameters accordingly optimized. The overall trends in coverage of Termitomyces diversity and host associations were estimated using Genbank data. Results indicate a monoculture of Termitomyces in the guts as well as the isolation sources (fungal combs). However, cases of more than one Termitomyces strains per mound were observed since mounds can contain different termite colonies. The newly found cultures, as well as the clustering analysis of GenBank data indicate that there are on average between one and two host genera per Termitomyces species. Saturation does not appear to have been reached, neither for the total number of known Termitomyces species nor for the number of Termitomyces species per host taxon, nor for the number of known hosts per Termitomyces species. Considering the rarity of Termitomyces fruiting bodies, it is suggested to base the future taxonomy of the group mainly on well

  16. Diversity of Termitomyces associated with fungus-farming termites assessed by cultural and culture-independent methods.

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    Huxley M Makonde

    Full Text Available BACKGROUND: Fungus-cultivating termites make use of an obligate mutualism with fungi from the genus Termitomyces, which are acquired through either vertical transmission via reproductive alates or horizontally transmitted during the formation of new mounds. Termitomyces taxonomy, and thus estimating diversity and host specificity of these fungi, is challenging because fruiting bodies are rarely found. Molecular techniques can be applied but need not necessarily yield the same outcome than morphological identification. METHODOLOGY: Culture-dependent and culture-independent methods were used to comprehensively assess host specificity and gut fungal diversity. Termites were identified using mitochondrial cytochrome oxidase II (COII genes. Twenty-three Termitomyces cultures were isolated from fungal combs. Internal transcribed spacer (ITS clone libraries were constructed from termite guts. Presence of Termitomyces was confirmed using specific and universal primers. Termitomyces species boundaries were estimated by cross-comparison of macromorphological and sequence features, and ITS clustering parameters accordingly optimized. The overall trends in coverage of Termitomyces diversity and host associations were estimated using Genbank data. RESULTS AND CONCLUSION: Results indicate a monoculture of Termitomyces in the guts as well as the isolation sources (fungal combs. However, cases of more than one Termitomyces strains per mound were observed since mounds can contain different termite colonies. The newly found cultures, as well as the clustering analysis of GenBank data indicate that there are on average between one and two host genera per Termitomyces species. Saturation does not appear to have been reached, neither for the total number of known Termitomyces species nor for the number of Termitomyces species per host taxon, nor for the number of known hosts per Termitomyces species. Considering the rarity of Termitomyces fruiting bodies, it is

  17. Comparison of Standard Culture-Based Method to Culture-Independent Method for Evaluation of Hygiene Effects on the Hand Microbiome.

    Science.gov (United States)

    Zapka, C; Leff, J; Henley, J; Tittl, J; De Nardo, E; Butler, M; Griggs, R; Fierer, N; Edmonds-Wilson, S

    2017-03-28

    Hands play a critical role in the transmission of microbiota on one's own body, between individuals, and on environmental surfaces. Effectively measuring the composition of the hand microbiome is important to hand hygiene science, which has implications for human health. Hand hygiene products are evaluated using standard culture-based methods, but standard test methods for culture-independent microbiome characterization are lacking. We sampled the hands of 50 participants using swab-based and glove-based methods prior to and following four hand hygiene treatments (using a nonantimicrobial hand wash, alcohol-based hand sanitizer [ABHS], a 70% ethanol solution, or tap water). We compared results among culture plate counts, 16S rRNA gene sequencing of DNA extracted directly from hands, and sequencing of DNA extracted from culture plates. Glove-based sampling yielded higher numbers of unique operational taxonomic units (OTUs) but had less diversity in bacterial community composition than swab-based sampling. We detected treatment-induced changes in diversity only by using swab-based samples (P hand hygiene industry methods and for future hand microbiome studies. On the basis of our results and previously published studies, we propose recommendations for best practices in hand microbiome research.IMPORTANCE The hand microbiome is a critical area of research for diverse fields, such as public health and forensics. The suitability of culture-independent methods for assessing effects of hygiene products on microbiota has not been demonstrated. This is the first controlled laboratory clinical hand study to have compared traditional hand hygiene test methods with newer culture-independent characterization methods typically used by skin microbiologists. This study resulted in recommendations for hand hygiene product testing, development of methods, and future hand skin microbiome research. It also demonstrated the importance of inclusion of skin physiological metadata in skin

  18. Identification of bacteria associated with feline chronic gingivostomatitis using culture-dependent and culture-independent methods.

    Science.gov (United States)

    Dolieslager, Sanne M J; Riggio, Marcello P; Lennon, Alan; Lappin, David F; Johnston, Norman; Taylor, David; Bennett, David

    2011-02-24

    Feline chronic gingivostomatitis (FCGS) is a chronic inflammatory disease of the oral cavity that causes severe pain and distress. There are currently no specific treatment methods available and little is known regarding its aetiology, although bacteria are thought to play a major role. The purpose of this study was to identify the oral bacterial flora in normal and diseased cats. Oral swabs were obtained from the palatoglossal folds of eight cats (three normal and five FCGS) and were subjected to microbiological culture. Pasteurella pneumotropica and Pasteurella multocida subsp. multocida were the most prevalent species identified by culture methods in the normal and FCGS samples, respectively. Bacteria were also identified using culture-independent methods (bacterial 16S rRNA gene sequencing). For the normal samples, 158 clones were analysed and 85 clones were sequenced. Capnocytophaga canimorsus (10.8% of clones analysed) was the predominant species. Uncultured species accounted for 8.2% of clones analysed, and 43.7% of clones analysed represented potentially novel species. For the FCGS samples, 253 clones were analysed and 91 clones were sequenced. The predominant species was P. multocida subsp. multocida (51.8% of clones analysed). Uncultured species accounted for 8.7% of clones analysed, and 4.7% of clones analysed represented potentially novel species. It is concluded that the oral flora in cats with FCGS appears to be less diverse than that found in normal cats. However, P. multocida subsp. multocida is found to be significantly more prevalent in FCGS than in normal cats and consequently may be of aetiological significance in this disease. Copyright © 2010 Elsevier B.V. All rights reserved.

  19. The bacteriology of chronic venous leg ulcer examined by culture-independent molecular methods

    DEFF Research Database (Denmark)

    Thomsen, Trine R; Aasholm, Martin S; Rudkjøbing, Vibeke B;

    2010-01-01

    , such as 16S rRNA gene sequencing, fingerprinting, quantitative polymerase chain reaction, and fluorescence in situ hybridization. Each wound contained an average of 5.4 species but the actual species varied between wounds. The diversity determined by culture-based methods and the molecular biological methods...... was different. All the wounds contained Staphylococcus aureus, whereas Pseudomonas aeruginosa was in six out of 14 wounds. Molecular methods detected anaerobic pathogens in four ulcers that were not detected with anaerobic culture methods. Quantitative polymerase chain reaction was used to compare the abundance......The bacterial microbiota plays an important role in the prolonged healing of chronic venous leg ulcers. The present study compared the bacterial diversity within ulcer material from 14 skin graft operations of chronic venous leg ulcers using culture-based methods and molecular biological methods...

  20. Application of culture culture-independent molecular biology based methods to evaluate acetic acid bacteria diversity during vinegar processing.

    Science.gov (United States)

    Ilabaca, Carolina; Navarrete, Paola; Mardones, Pamela; Romero, Jaime; Mas, Albert

    2008-08-15

    Acetic acid bacteria (AAB) are considered fastidious microorganisms because they are difficult to isolate and cultivate. Different molecular approaches were taken to detect AAB diversity, independently of their capacity to grow in culture media. Those methods were tested in samples that originated during traditional vinegar production. Bacterial diversity was assessed by analysis of 16S rRNA gene, obtained by PCR amplifications of DNA extracted directly from the acetification container. Bacterial composition was analyzed by RFLP-PCR of 16S rRNA gene, Temporal Temperature Gradient Gel Electrophoresis (TTGE) separation of amplicons containing region V3-V5 of 16S rRNA gene and cloning of those amplicons. TTGE bands and clones were grouped based on their electrophoretic pattern similarity and sequenced to be compared with reference strains. The main microorganism identified in vinegar was Acetobacter pasteurianus, which at the end of the acetification process was considered to be the only microorganism present. The diversity was the highest at 2% acetic acid, where indefinite species of Gluconacetobacter xylinus/europaeus/intermedius were also present.

  1. Microbial diversity of a Camembert-type cheese using freeze-dried Tibetan kefir coculture as starter culture by culture-dependent and culture-independent methods.

    Science.gov (United States)

    Mei, Jun; Guo, Qizhen; Wu, Yan; Li, Yunfei

    2014-01-01

    The biochemical changes occurring during cheese ripening are directly and indirectly dependent on the microbial associations of starter cultures. Freeze-dried Tibetan kefir coculture was used as a starter culture in the Camembert-type cheese production for the first time. Therefore, it's necessary to elucidate the stability, organization and identification of the dominant microbiota presented in the cheese. Bacteria and yeasts were subjected to culture-dependent on selective media and culture-independent polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) analysis and sequencing of dominant bands to assess the microbial structure and dynamics through ripening. In further studies, kefir grains were observed using scanning electron microscopy (SEM) methods. A total of 147 bacteria and 129 yeasts were obtained from the cheese during ripening. Lactobacillus paracasei represents the most commonly identified lactic acid bacteria isolates, with 59 of a total of 147 isolates, followed by Lactococcus lactis (29 isolates). Meanwhile, Kazachstania servazzii (51 isolates) represented the mainly identified yeast isolate, followed by Saccharomyces cerevisiae (40 isolates). However, some lactic acid bacteria detected by sequence analysis of DGGE bands were not recovered by plating. The yeast S. cerevisiae and K. servazzii are described for the first time with kefir starter culture. SEM showed that the microbiota were dominated by a variety of lactobacilli (long and curved) cells growing in close association with a few yeasts in the inner portion of the grain and the short lactobacilli were observed along with yeast cells on the exterior portion. Results indicated that conventional culture method and PCR-DGGE should be combined to describe in maximal detail the microbiological composition in the cheese during ripening. The data could help in the selection of appropriate commercial starters for Camembert-type cheese.

  2. Microbial diversity of a Camembert-type cheese using freeze-dried Tibetan kefir coculture as starter culture by culture-dependent and culture-independent methods.

    Directory of Open Access Journals (Sweden)

    Jun Mei

    Full Text Available The biochemical changes occurring during cheese ripening are directly and indirectly dependent on the microbial associations of starter cultures. Freeze-dried Tibetan kefir coculture was used as a starter culture in the Camembert-type cheese production for the first time. Therefore, it's necessary to elucidate the stability, organization and identification of the dominant microbiota presented in the cheese. Bacteria and yeasts were subjected to culture-dependent on selective media and culture-independent polymerase chain reaction (PCR-denaturing gradient gel electrophoresis (DGGE analysis and sequencing of dominant bands to assess the microbial structure and dynamics through ripening. In further studies, kefir grains were observed using scanning electron microscopy (SEM methods. A total of 147 bacteria and 129 yeasts were obtained from the cheese during ripening. Lactobacillus paracasei represents the most commonly identified lactic acid bacteria isolates, with 59 of a total of 147 isolates, followed by Lactococcus lactis (29 isolates. Meanwhile, Kazachstania servazzii (51 isolates represented the mainly identified yeast isolate, followed by Saccharomyces cerevisiae (40 isolates. However, some lactic acid bacteria detected by sequence analysis of DGGE bands were not recovered by plating. The yeast S. cerevisiae and K. servazzii are described for the first time with kefir starter culture. SEM showed that the microbiota were dominated by a variety of lactobacilli (long and curved cells growing in close association with a few yeasts in the inner portion of the grain and the short lactobacilli were observed along with yeast cells on the exterior portion. Results indicated that conventional culture method and PCR-DGGE should be combined to describe in maximal detail the microbiological composition in the cheese during ripening. The data could help in the selection of appropriate commercial starters for Camembert-type cheese.

  3. Microbial Diversity of a Camembert-Type Cheese Using Freeze-Dried Tibetan Kefir Coculture as Starter Culture by Culture-Dependent and Culture-Independent Methods

    Science.gov (United States)

    Mei, Jun; Guo, Qizhen; Wu, Yan; Li, Yunfei

    2014-01-01

    The biochemical changes occurring during cheese ripening are directly and indirectly dependent on the microbial associations of starter cultures. Freeze-dried Tibetan kefir coculture was used as a starter culture in the Camembert-type cheese production for the first time. Therefore, it's necessary to elucidate the stability, organization and identification of the dominant microbiota presented in the cheese. Bacteria and yeasts were subjected to culture-dependent on selective media and culture-independent polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) analysis and sequencing of dominant bands to assess the microbial structure and dynamics through ripening. In further studies, kefir grains were observed using scanning electron microscopy (SEM) methods. A total of 147 bacteria and 129 yeasts were obtained from the cheese during ripening. Lactobacillus paracasei represents the most commonly identified lactic acid bacteria isolates, with 59 of a total of 147 isolates, followed by Lactococcus lactis (29 isolates). Meanwhile, Kazachstania servazzii (51 isolates) represented the mainly identified yeast isolate, followed by Saccharomyces cerevisiae (40 isolates). However, some lactic acid bacteria detected by sequence analysis of DGGE bands were not recovered by plating. The yeast S. cerevisiae and K. servazzii are described for the first time with kefir starter culture. SEM showed that the microbiota were dominated by a variety of lactobacilli (long and curved) cells growing in close association with a few yeasts in the inner portion of the grain and the short lactobacilli were observed along with yeast cells on the exterior portion. Results indicated that conventional culture method and PCR-DGGE should be combined to describe in maximal detail the microbiological composition in the cheese during ripening. The data could help in the selection of appropriate commercial starters for Camembert-type cheese. PMID:25360757

  4. Microbial community analysis of food-spoilage bacteria in commercial custard creams using culture-dependent and independent methods.

    Science.gov (United States)

    Arakawa, K; Kawai, Y; Iioka, H; Tanioka, M; Nishimura, J; Kitazawa, H; Tsurumi, K; Saito, T

    2008-08-01

    Custard cream is made from highly nutritive raw materials such as milk and sugar and is easily spoiled by the multiplication of specific microbial contaminants or residents. However, this spoilage microbial community has not been studied. We determined the spoilage microbiota in commercial custard creams using culture-dependent and independent methods. Using the culture-dependent analysis with various agar media, 185 bacterial colonies and 43 eukaryal colonies were isolated from 7 commercial custard cream products. All bacterial isolates were morphologically, physiologically, and genetically identified as bacilli, staphylococci, lactic acid bacteria, and psychrotrophic gram-negative rods. Using culture-independent molecular analysis, the PCR-denaturing gradient gel electrophoresis technique, spoilage of the commercial custard creams was found to be caused by bacilli, staphylococci, lactic acid bacteria, psychrotrophic gram-negative rods, Anoxybacillus sp., Caurobacter sp., and Streptococcus sp. bacteria. The detected spoilage bacteria were the same species as previously detected in spoiled milk products and shown in other reports, suggesting that spoilage bacteria in a raw material easily grow in processed foods made from milk. We determined the spoilage microbial communities in commercial custard creams, and these are the first data concerning spoilage microbiota in nonfermented processed foods using a culture-independent analysis. Our study will be useful for the manufacture and safe preservation of dairy products because the first step toward safe food preservation by food manufacturers is to understand the spoilage microbiota in a target food to select optimal preservatives and to reduce the use of food additives.

  5. ADSA Foundation Scholar Award: Trends in culture-independent methods for assessing dairy food quality and safety: emerging metagenomic tools.

    Science.gov (United States)

    Yeung, Marie

    2012-12-01

    Enhancing the quality and safety of dairy food is critical to maintaining the competitiveness of dairy products in the food and beverage market and in reinforcing consumer confidence in the dairy industry. Raw milk quality has a significant effect on finished product quality. Several microbial groups found in raw milk have been shown to adversely affect the shelf life of pasteurized milk. Current microbiological criteria used to define milk quality are based primarily on culture-dependent methods, some of which are perceived to lack the desired sensitivity and specificity. To supplement traditional methods, culture-independent methods are increasingly being used to identify specific species or microbial groups, and to detect indicator genes or proteins in raw milk or dairy products. Some molecular subtyping techniques have been developed to track the transmission of microbes in dairy environments. The burgeoning "-omics" technologies offer new and exciting opportunities to enhance our understanding of food quality and safety in relation to microbes. Metagenomics has the potential to characterize microbial diversity, detect nonculturable microbes, and identify unique sequences or other factors associated with dairy product quality and safety. In this review, fluid milk will be used as the primary example to examine the adequacy and validity of conventional methods, the current trend of culture-independent methods, and the potential applications of metagenomics in dairy food research.

  6. Identification, isolation and quantification of representative bacteria from fermented cassava dough using an integrated approach of culture-dependent and culture-independent methods.

    Science.gov (United States)

    Miambi, Edouard; Guyot, Jean-Pierre; Ampe, Frédéric

    2003-04-25

    The use of denaturing gradient gel electrophoresis (DGGE) and traditional culture-depending methods for examining the bacterial community of traditional cassava starch fermentation were investigated. It appeared that DGGE profiles of total DNA of cassava dough exhibited 10 distinguishable bands. In contrast, DGGE fingerprints of bacteria recovered from enrichment cultures of fermented dough gave variable profiles containing fewer bands. Bands corresponding to five bacterial species detected by direct PCR-DGGE of total DNA from of cassava dough were also observed in DGGE patterns of enrichment cultures. Eighteen strains were isolated from cultures selected on the basis of their DGGE banding patterns. Assessment of bacterial identification by 16S rDNA sequence similarity revealed that band comigration implied sequence identity. Comparison of 16S rDNA sequences of excised DGGE bands and recovered pure culture isolates with those in GENBANK and the RDP databases revealed that representative bacteria of fermented cassava dough were Lactobacillus and Pediococcus species as well as species of Clostridium, Propionibacterium and Bacillus. Some Lactobacillus species detected in dough samples by sequence analysis of DGGE bands were not recovered in any of the five culture media and conditions used. On the other hand, some species recovered as pure cultures from enrichments were not detected by direct DGGE analysis of total bacterial DNA from cassava dough. Our results provide evidence of the necessity to combine both culture-dependent and culture-independent methods for better description of microbial communities in indigenous cassava starch fermentations.

  7. Culture-Dependent and -Independent Methods to Investigate the Predominant Microorganisms Associated with Wet Processed Coffee.

    Science.gov (United States)

    Feng, Xiaomin; Dong, Honghong; Yang, Pan; Yang, Ruijuan; Lu, Jun; Lv, Jie; Sheng, Jun

    2016-08-01

    The fermentation process of Yunnan arabica coffee is a typical wet fermentation. Its excellent quality is closely related to microbes in the process of fermentation. The purpose of this study was to isolate and identify the microorganisms in the wet method of coffee processing in Yunnan Province, China. Microbial community structure and dominant bacterial species were evaluated by traditional cultivated separation method and PCR-DGGE technology, and were further analyzed in combination with the changes of organic acid content, activity of pectinase, and physical parameters (pH and temperature). A large number of microorganisms which can produce pectinase were found. Among them, Enterobacter cowanii, Pantoea agglomerans, Enterobacteriaceae bacterium, and Rahnella aquatilis were the predominant gram-negative bacteria, Bacillus cereus was the predominant gram-positive bacterium, Pichia kluyveri, Hanseniaspora uvarum, and Pichia fermentans were the predominant yeasts, and all those are pectinase-producing microorganisms. As for the contents of organic acids, oxalic was the highest, followed by acetic and lactic acids. Butyrate and propionate, which were unfavorable during the fermentation period, were barely discovered.

  8. Culture-independent methods to study subaerial biofilm growing on biodeteriorated surfaces of stone cultural heritage and frescoes.

    Science.gov (United States)

    Cappitelli, Francesca; Villa, Federica; Polo, Andrea

    2014-01-01

    Actinobacteria, cyanobacteria, algae, and fungi form subaerial biofilm (SAB) that can lead to material deterioration on artistic stone and frescoes. In studying SAB on cultural heritage surfaces, a general approach is to combine microscopy observations and molecular analyses. Sampling of biofilm is performed using specific adhesive tape and sampling of SAB and the substrate with sterile scalpels and chisels. Biofilm observations are carried out using optical and scanning electron microscopy. Specific taxa and EPS in biofilm can be readily visualized by fluorochrome staining and subsequent observation using fluorescence or confocal laser scanning microscopy. The observation of cross sections containing both SAB and the substrate shows if biofilm has developed not only on the surface but also underneath. Following nucleic acid extraction, 16S rRNA gene sequencing is used to identify bacterial taxa, while 18S rRNA gene and internal transcribed spacer (ITS) sequence analysis is used to study eukaryotic groups. In this chapter, we illustrate the protocols related to fluorescence in situ hybridization (FISH), scanning electron microscopy (SEM), and denaturing gradient gel electrophoresis (DGGE).

  9. Combination of culture-independent and culture-dependent molecular methods for the determination of bacterial community of iru, a fermented Parkia biglobosa seeds.

    Science.gov (United States)

    Adewumi, Gbenga A; Oguntoyinbo, Folarin A; Keisam, Santosh; Romi, Wahengbam; Jeyaram, Kumaraswamy

    2012-01-01

    In this study, bacterial composition of iru produced by natural, uncontrolled fermentation of Parkia biglobosa seeds was assessed using culture-independent method in combination with culture-based genotypic typing techniques. PCR-denaturing gradient gel electrophoresis (DGGE) revealed similarity in DNA fragments with the two DNA extraction methods used and confirmed bacterial diversity in the 16 iru samples from different production regions. DNA sequencing of the highly variable V3 region of the 16S rRNA genes obtained from PCR-DGGE identified species related to Bacillus subtilis as consistent bacterial species in the fermented samples, while other major bands were identified as close relatives of Staphylococcus vitulinus, Morganella morganii, B. thuringiensis, S. saprophyticus, Tetragenococcus halophilus, Ureibacillus thermosphaericus, Brevibacillus parabrevis, Salinicoccus jeotgali, Brevibacterium sp. and uncultured bacteria clones. Bacillus species were cultured as potential starter cultures and clonal relationship of different isolates determined using amplified ribosomal DNA restriction analysis (ARDRA) combined with 16S-23S rRNA gene internal transcribed spacer (ITS) PCR amplification, restriction analysis (ITS-PCR-RFLP), and randomly amplified polymorphic DNA (RAPD-PCR). This further discriminated B. subtilis and its variants from food-borne pathogens such as B. cereus and suggested the need for development of controlled fermentation processes and good manufacturing practices (GMP) for iru production to achieve product consistency, safety quality, and improved shelf life.

  10. Combination of culture-independent and culture-dependent molecular methods for the determination of bacterial community of iru, a fermented Parkia biglobosa seeds.

    Directory of Open Access Journals (Sweden)

    Gbenga Adedeji Adewumi

    2013-01-01

    Full Text Available In this study, bacterial composition of iru produced by natural, uncontrolled fermentation of Parkia biglobosa seeds was assessed using culture-independent method in combination with culture-based genotypic typing techniques. PCR-denaturing gradient gel electrophoresis (DGGE revealed similarity in DNA fragments with the two DNA extraction methods used and confirmed bacterial diversity in the sixteen iru samples from different production regions. DNA sequencing of the highly variable V3 region of the 16S rRNA genes obtained from PCR-DGGE identified species related to Bacillus subtilis as consistent bacterial species in the fermented samples, while other major bands were identified as close relatives of Staphylococcus vitulinus, Morganella morganii, B. thuringiensis, Staphylococcus saprophyticus, Tetragenococcus halophilus, Ureibacillus thermosphaericus, Brevibacillus parabrevis, Salinicoccus jeotgali, Brevibacterium sp. and Uncultured bacteria clones. Bacillus species were cultured as potential starter cultures and clonal relationship of different isolates determined using amplified ribosomal DNA restriction analysis (ARDRA combined with 16S-23S rRNA gene internal transcribed spacer (ITS PCR amplification, restriction analysis (ITS-PCR-RFLP and randomly amplified polymorphic DNA (RAPD-PCR. This further discriminated Bacillus subtilis and its variants from food-borne pathogens such as Bacillus cereus and suggested the need for development of controlled fermentation processes and good manufacturing practices (GMP for iru production to achieve product consistency, safety quality and improved shelf life.

  11. Identification of bacteria associated with feline chronic gingivostomatitis using culture-dependent and culture-independent methods

    OpenAIRE

    Dolieslager, Sanne M.J.; Riggio, Marcello P; Lennon, Alan; Lappin, David F; Johnston, Norman; Taylor, David; Bennett, David

    2011-01-01

    Abstract Feline chronic gingivostomatitis (FCGS) is a chronic inflammatory disease of the oral cavity that causes severe pain and distress. There are currently no specific treatment methods available and little is known regarding its aetiology, although bacteria are thought to play a major role. The purpose of this study was to identify the oral bacterial flora in normal and diseased cats. Oral swabs were obtained from the palatoglossal folds of eight cats (three normal, five FCGS)...

  12. Characterization of the Bacterial Community Associated with Larvae and Adults of Anoplophora chinensis Collected in Italy by Culture and Culture-Independent Methods

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    Aurora Rizzi

    2013-01-01

    Full Text Available The wood-boring beetle Anoplophora chinensis Forster, native to China, has recently spread to North America and Europe causing serious damage to ornamental and forest trees. The gut microbial community associated with these xylophagous beetles is of interest for potential biotechnological applications in lignocellulose degradation and development of pest-control measures. In this study the gut bacterial community of larvae and adults of A. chinensis, collected from different host trees in North Italy, was investigated by both culture and culture-independent methods. Larvae and adults harboured a moderately diverse bacterial community, dominated by Proteobacteria, Actinobacteria, and Firmicutes. The gammaproteobacterial family Enterobacteriaceae (genera Gibbsiella, Enterobacter, Raoultella, and Klebsiella was the best represented. The abundance of such bacteria in the insect gut is likely due to the various metabolic abilities of Enterobacteriaceae, including fermentation of carbohydrates derived from lignocellulose degradation and contribution to nitrogen intake by nitrogen-fixing activity. In addition, bacteria previously shown to have some lignocellulose-degrading activity were detected at a relatively low level in the gut. These bacteria possibly act synergistically with endogenous and fungal enzymes in lignocellulose breakdown. The detection of actinobacterial symbionts could be explained by a possible role in the detoxification of secondary plant metabolites and/or protection against pathogens.

  13. Culture-dependent and culture-independent methods for molecular analysis of the diversity of lactobacilli in "Camembert de Normandie" cheese

    OpenAIRE

    Henri-Dubernet, Ségolène; Desmasures, Nathalie; Guéguen, Micheline

    2004-01-01

    International audience; We compared a culture-dependent and a culture-independent approach for the assessment of lactobacilli community biodiversity and evolution during the production of RDO Camembert in three cheese-making factories. We used temperature gradient gel electrophoresis (TGGE) to analyse total microbial DNA and DNA from single isolates. TGGE patterns of total microbial DNA from milk and cheese showed that Lactobacillus paracasei subsp. paracasei was a dominant species in the thr...

  14. Molecular profiling of fungal communities in moisture damaged buildings before and after remediation - a comparison of culture-dependent and culture-independent methods

    Directory of Open Access Journals (Sweden)

    Auvinen Petri

    2011-10-01

    Full Text Available Abstract Background Indoor microbial contamination due to excess moisture is an important contributor to human illness in both residential and occupational settings. However, the census of microorganisms in the indoor environment is limited by the use of selective, culture-based detection techniques. By using clone library sequencing of full-length internal transcribed spacer region combined with quantitative polymerase chain reaction (qPCR for 69 fungal species or assay groups and cultivation, we have been able to generate a more comprehensive description of the total indoor mycoflora. Using this suite of methods, we assessed the impact of moisture damage on the fungal community composition of settled dust and building material samples (n = 8 and 16, correspondingly. Water-damaged buildings (n = 2 were examined pre- and post- remediation, and compared with undamaged reference buildings (n = 2. Results Culture-dependent and independent methods were consistent in the dominant fungal taxa in dust, but sequencing revealed a five to ten times higher diversity at the genus level than culture or qPCR. Previously unknown, verified fungal phylotypes were detected in dust, accounting for 12% of all diversity. Fungal diversity, especially within classes Dothideomycetes and Agaricomycetes tended to be higher in the water damaged buildings. Fungal phylotypes detected in building materials were present in dust samples, but their proportion of total fungi was similar for damaged and reference buildings. The quantitative correlation between clone library phylotype frequencies and qPCR counts was moderate (r = 0.59, p Conclusions We examined a small number of target buildings and found indications of elevated fungal diversity associated with water damage. Some of the fungi in dust were attributable to building growth, but more information on the material-associated communities is needed in order to understand the dynamics of microbial communities between

  15. Identification and characterization of lactic acid bacteria and yeasts of PDO Tuscan bread sourdough by culture dependent and independent methods.

    Science.gov (United States)

    Palla, Michela; Cristani, Caterina; Giovannetti, Manuela; Agnolucci, Monica

    2017-06-05

    Sourdough fermentation has been increasingly used worldwide, in accordance with the demand of consumers for tasty, natural and healthy food. The high diversity of lactic acid bacteria (LAB) and yeast species, detected in sourdoughs all over the world, may affect nutritional, organoleptic and technological traits of leavened baked goods. A wide regional variety of traditional sourdough breads, over 200 types, has been recorded in Italy, including special types selected as worthy of either Protected Geographical Indication (PGI) or Protected Designation of Origin (PDO), whose sourdough microbiota has been functionally and molecularly characterized. As, due to the very recent designation, the microbiota of Tuscan bread sourdough has not been investigated so far, the aim of the present work was to isolate and characterize the species composition of LAB and yeasts of PDO Tuscan bread sourdough by culture-independent and dependent methods. A total of 130 yeasts from WLN medium and 193 LAB from both mMRS and SDB media were isolated and maintained to constitute the germplasm bank of PDO Tuscan bread. Ninety six LAB from mMRS medium and 68 yeasts from WLN medium were randomly selected and molecularly identified by ARDRA (Amplified Ribosomal DNA Restriction Analysis) and PCR-RFLP analysis of the ITS region, respectively, and sequencing. The yeast identity was confirmed by 26S D1/D2 sequencing. All bacterial isolates showed 99% identity with Lactobacillus sanfranciscensis, 65 yeast isolates were identified as Candida milleri, and 3 as Saccharomyces cerevisiae. Molecular characterization of PDO Tuscan bread sourdough by PCR-DGGE confirmed such data. The distinctive tripartite species association, detected as the microbiota characterizing the sourdough used to produce PDO Tuscan bread, encompassed a large number of L. sanfranciscensis and C. milleri strains, along with a few of S. cerevisiae. The relative composition and specific physiological characteristics of such microbiota

  16. Evaluation of microbial diversity in sulfite-added and sulfite-free wine by culture-dependent and -independent methods.

    Science.gov (United States)

    Takahashi, Masayuki; Ohta, Tami; Masaki, Kazuo; Mizuno, Akihiro; Goto-Yamamoto, Nami

    2014-05-01

    The difference in microbiota including non-lactic acid bacteria, non-acetic acid bacteria, and wild yeast during winemaking and in the end-products between sulfite-added and sulfite-free wine, was investigated using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and a culture-dependent method. There were differences between the microorganisms detected by PCR-DGGE and those detected by the culture-dependent method, probably because of the selectivity of culture medium and the characteristics of PCR-based method. In both the red wine and white wine, the microbial diversity of the sulfite-added wine was lower than that of the sulfite-free wine during fermentation. Tatumella terrea was detected from the fermenting must by PCR-DGGE and by the culture-dependent method, even though sulfite inhibited its growth to some extent. We confirmed that the addition of sulfite plays an important role in winemaking by inhibiting the growth of unexpected microorganisms, but on the other hand, it was revealed that some microorganisms can survive and grow in sulfite-added fermenting must. We also analyzed 15 samples of commercial wines by the PCR-DGGE method and detected various microorganisms. Among them, Sphingomonas sp., Pseudozyma sp., Ochromonas sp. and Methylophilus sp. were found for the first time in wine as far as we know. We did not identify a specific microorganism that was detected only from wines without sulfite addition. Thus, the microbiota of end-products seemed to be influenced by other factors, such as filtration before bottling, the production equipment and the storage environment. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  17. Study of microbial diversity in raw milk and fresh curd used for Fontina cheese production by culture-independent methods.

    Science.gov (United States)

    Giannino, Maria Laura; Marzotto, Marta; Dellaglio, Franco; Feligini, Maria

    2009-04-15

    The bacterial populations of raw milk employed for the production of Fontina cheese in alpine farms located in different valleys and altitudes (from 700 to 2246 m above sea level) were investigated by culture independent techniques. Total microbial DNA was isolated from milk and curd samples and used as template in Polymerase Chain Reaction (PCR) to study the hypervariable V3 region of the bacterial 16S rRNA gene and analyzed by Denaturing Gradient Gel Electrophoresis (DGGE). Representative bands of DGGE patterns were sequenced for identification purposes. The use of universal primer for PCR-DGGE allowed the description of the bacterial community, not only for the presence of lactic acid bacteria, but also for other adventitious species. DGGE profiles obtained from milk and fresh curd samples were generally different and typical for each farm, although some recurrent bands were observed. Cluster analysis of DGGE profiles did not show high similarity among samples and it was probably dependent on the different geographical areas of pastures. Some Lactic Acid Bacteria (LAB) recurred in many samples (Streptococcus thermophilus, Enterococcus faecium, Enterococcus faecalis, Lactococcus lactis, Leuconostoc lactis) indicating that alpine milk is a preferential niche for their colonization. The microbiota included not only mesophilic and thermoresistant LAB but also adventitious bacteria (Macrococcus caseolyticus, Rothia spp.) and psychrotrophic bacteria (Chryseobacterium spp., Pseudomonas spp.), that were found in almost all samples, but disappeared after the warming up at 47-48 degrees C of coagulated milk. Pantoea spp. was primarily found in curds and only with a low incidence in milk samples, indicating the environmental origin. Finally the sequencing data confirmed the presence of E. faecium, E. faecalis and S. thermophilus as major species present in the curd. These species were found also in raw milk, proving its importance as source of the typical fermenting

  18. Polyphasic approach to bacterial dynamics during the ripening of Spanish farmhouse cheese, using culture-dependent and -independent methods.

    Science.gov (United States)

    Martín-Platero, Antonio M; Valdivia, Eva; Maqueda, Mercedes; Martín-Sánchez, Inés; Martínez-Bueno, Manuel

    2008-09-01

    We studied the dynamics of the microbial population during ripening of Cueva de la Magahá cheese using a combination of classical and molecular techniques. Samples taken during ripening of this Spanish goat's milk cheese in which Lactococcus lactis and Streptococcus thermophilus were used as starter cultures were analyzed. All bacterial isolates were clustered by using randomly amplified polymorphic DNA (RAPD) and identified by 16S rRNA gene sequencing, species-specific PCR, and multiplex PCR. Our results indicate that the majority of the 225 strains isolated and enumerated on solid media during the ripening period were nonstarter lactic acid bacteria, and Lactobacillus paracasei was the most abundant species. Other Lactobacillus species, such as Lactobacillus plantarum and Lactobacillus parabuchneri, were also detected at the beginning and end of ripening, respectively. Non-lactic-acid bacteria, mainly Kocuria and Staphylococcus strains, were also detected at the end of the ripening period. Microbial community dynamics determined by temporal temperature gradient gel electrophoresis provided a more precise estimate of the distribution of bacteria and enabled us to detect Lactobacillus curvatus and the starter bacteria S. thermophilus and L. lactis, which were not isolated. Surprisingly, the bacterium most frequently found using culture-dependent analysis, L. paracasei, was scarcely detected by this molecular approach. Finally, we studied the composition of the lactobacilli and their evolution by using length heterogeneity PCR.

  19. Complex microbiota of a Chinese "Fen" liquor fermentation starter (Fen-Daqu), revealed by culture-dependent and culture-independent methods

    NARCIS (Netherlands)

    Zheng, X.; Zheng, Y.; Han, B.; Zwietering, M.H.; Samson, R.A.; Boekhout, T.; Nout, M.J.R.

    2012-01-01

    Daqu is a traditional fermentation starter that is used for Chinese liquor production. Although partly mechanized, its manufacturing process has remained traditional. We investigated the microbial diversity of Fen-Daqu, a starter for light-flavour liquor, using combined culture-dependent and culture

  20. Complex microbiota of a Chinese "Fen" liquor fermentation starter (Fen-Daqu), revealed by culture-dependent and culture-independent methods

    NARCIS (Netherlands)

    Zheng, X.W.; Yan, Z.; Han, B.Z.; Zwietering, M.H.; Samson, R.A.; Boekhout, T.; Nout, M.J.R.

    2012-01-01

    Daqu is a traditional fermentation starter that is used for Chinese liquor production. Although partly mechanized, its manufacturing process has remained traditional. We investigated the microbial diversity of Fen- Daqu, a starter for light-flavour liquor, using combined culture-dependent and cultur

  1. Application of culture-dependent and culture-independent methods for the identification of Lactobacillus kefiranofaciens in microbial consortia present in kefir grains.

    Science.gov (United States)

    Hamet, Maria Fernanda; Londero, Alejandra; Medrano, Micaela; Vercammen, Elisabeth; Van Hoorde, Koenraad; Garrote, Graciela L; Huys, Geert; Vandamme, Peter; Abraham, Analía G

    2013-12-01

    The biological and technological characteristics of kefiran as well as its importance in grain integrity led us to analyze the microbial kefir grain consortium with focus on Lactobacillus kefiranofaciens. The presence of L. kefiranofaciens in the nine kefir grains studied was demonstrated by denaturing gradient gel electrophoresis. By culture dependent methods applying a methodology focused on the search of this species, 22 isolates with typical morphology were obtained and identified applying a combination of SDS-PAGE of whole cell proteins, (GTG)5-PCR and sequence analysis of the housekeeping gene encoding the α-subunit of bacterial phenylalanyl-tRNA synthase (pheS). This polyphasic approach allowed the reliable identification of 11 L. kefiranofaciens, 5 Lactobacillus paracasei, 4 Lactobacillus kefiri and 2 Lactobacillus parakefiri isolates. Isolated L. kefiranofaciens strains produced polysaccharide in strain-dependent concentrations and EPS produced by them also differed in the degree of polymerization. The isolation and accurate identification of L. kefiranofaciens is relevant taking into account the important role of this microorganism in the grain ecosystem as well as its potential application as starter in food fermentations.

  2. Evaluation of microbial diversity in the pilot-scale beer brewing process by culture-dependent and culture-independent method.

    Science.gov (United States)

    Takahashi, M; Kita, Y; Kusaka, K; Mizuno, A; Goto-Yamamoto, N

    2015-02-01

    In the brewing industry, microbial management is very important for stabilizing the quality of the product. We investigated the detailed microbial community of beer during fermentation and maturation, to manage beer microbiology in more detail. We brewed a beer (all-malt) and two beerlike beverages (half- and low-malt) in pilot-scale fermentation and investigated the microbial community of them using a next-generation sequencer (454 GS FLX titanium), quantitative PCR, flow cytometry and a culture-dependent method. From 28 to 88 genera of bacteria and from 9 to 38 genera of eukaryotic micro-organisms were detected in each sample. Almost all micro-organisms died out during the boiling process. However, bacteria belonging to the genera Acidovorax, Bacillus, Brevundimonas, Caulobacter, Chryseobacterium, Methylobacterium, Paenibacillus, Polaromonas, Pseudomonas, Ralstonia, Sphingomonas, Stenotrophomonas, Tepidimonas and Tissierella were detected at the early and middle stage of fermentation, even though their cell densities were low (below approx. 10(3) cells ml(-1) ) and they were not almost detected at the end of fermentation. We revealed that the microbial community of beer during fermentation and maturation is very diverse and several bacteria possibly survive during fermentation. In this study, we revealed the detailed microbial communities of beer using next-generation sequencing. Some of the micro-organisms detected in this study were found in beer brewing process for the first time. Additionally, the possibility of growth of several bacteria at the early and middle stage of fermentation was suggested. © 2014 The Society for Applied Microbiology.

  3. A procedure to evaluate the efficiency of surface sterilization methods in culture-independent fungal endophyte studies

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    R.J. Burgdorf

    2014-09-01

    Full Text Available Extraneous DNA interferes with PCR studies of endophytic fungi. A procedure was developed with which to evaluate the removal of extraneous DNA. Wheat (Triticum aestivum leaves were sprayed with Saccharomyces cerevisiae and then subjected to physical and chemical surface treatments. The fungal ITS1 products were amplified from whole tissue DNA extractions. ANOVA was performed on the DNA bands representing S. cerevisiae on the agarose gel. Band profile comparisons using permutational multivariate ANOVA (PERMANOVA and non-metric multidimensional scaling (NMDS were performed on DGGE gel data, and band numbers were compared between treatments. Leaf surfaces were viewed under variable pressure scanning electron microscopy (VPSEM. Yeast band analysis of the agarose gel showed that there was no significant difference in the mean band DNA quantity after physical and chemical treatments, but they both differed significantly (p < 0.05 from the untreated control. PERMANOVA revealed a significant difference between all treatments (p < 0.05. The mean similarity matrix showed that the physical treatment results were more reproducible than those from the chemical treatment results. The NMDS showed that the physical treatment was the most consistent. VPSEM indicated that the physical treatment was the most effective treatment to remove surface microbes and debris. The use of molecular and microscopy methods for the post-treatment detection of yeast inoculated onto wheat leaf surfaces demonstrated the effectiveness of the surface treatment employed, and this can assist researchers in optimizing their surface sterilization techniques in DNA-based fungal endophyte studies.

  4. Complex microbiota of a Chinese "Fen" liquor fermentation starter (Fen-Daqu), revealed by culture-dependent and culture-independent methods

    NARCIS (Netherlands)

    Zheng, X.W.; Yan, Z.; Han, B.Z.; Zwietering, M.H.; Samson, R.A.; Boekhout, T.; Nout, M.J.R.

    2012-01-01

    Daqu is a traditional fermentation starter that is used for Chinese liquor production. Although partly mechanized, its manufacturing process has remained traditional. We investigated the microbial diversity of Fen- Daqu, a starter for light-flavour liquor, using combined culture-dependent and

  5. Microbiology of shallow subsurface aquifer and carbonate rocks studied by culture-dependent and culture-independent methods. Preliminary results on an underground laboratory, the LSBB, Rustrel, France

    Directory of Open Access Journals (Sweden)

    Galès Grégoire

    2016-01-01

    Full Text Available We investigated the microbiology of a shallow subsurface site, the LSBB, located near Avignon. This site lies in carbonate rocks, belonging to the Urgonian facies. Rock, concrete and water samples were collected and directly transferred to the laboratory. Studies of microorganisms as pure cultures are the only way to get their real physiological properties. Nevertheless, microbiologists cannot cultivate and isolate the majority of microorganisms for several reasons, one being our lack of understanding of their minimal needs. Molecular studies, e.g. extraction and sequencing of the total nucleic acids present in an environment provide phylogenetic and metabolic information on uncultivated microorganisms. We performed aerobic and anaerobic culture with various electron acceptors and donors, searching for heterotrophic, methanogenic, sulphate- nitrate- and FeIII- reducing Prokaryotes. We also performed DNA extractions and PCR amplification of ribosomal RNA genes, to test if our protocols were adapted to this environment. Our results show that the LSBB galleries are colonized by a low diversity microbiote, with a strong influence of anthropogenic activities. Further studies will link the microorganisms biodiversity and the petrophysic properties of rocks.

  6. Corporate culture and participation: Independent constructs?

    Directory of Open Access Journals (Sweden)

    A. Odendaal

    1998-06-01

    Full Text Available The objective of this study was to determine whether participative management and corporate culture are mutually exclusive (independent concepts. The Human Sciences Research Council (HSRC used an organisational climate and culture questionnaire and applied it to the test sample of 2 357 personnel from a population of 23 564 people working for a paramilitary organisation. The data set of the HSRC was used for the purpose of this study. A panel of experts identified the items in the questionnaire which related to participation according to predetermined criteria and the remaining items were applicable to corporate culture. The results of the factor and item analyses indicate that participative management and corporate culture, as perceived by workers and management, were interrelated and were not independent concepts as conceptualised by the experts. Opsomming Die doel van hierdie studie is om te bepaal of deelnemende bestuur en korporatiewe kultuur onderling uitsluitende (onafhanklike konsepte is. Die Raad vir Geesteswetenskaplike Navorsing (RGN het 'n kultuur- en klimaatvraelys op 'n steekproef van 2 357 respondente, vanuit 'n populasie van 23 564 persone wat werksaam is in 'n para-militere instansie, toegepas. Die datastel van die RGN is vir die doel van hierdie studie gebruik. 'n Paneel deskundiges het dimensies wat op deelname betrekking het geidentifiseer aan die hand van voorafbepaalde kriteria en die oorblywende items was op kultuur van toepassing. Die resultate van die faktor- en itemontledings het bevind dat deelnemende bestuur en korporatiewe kultuur, soos bestudeer vanuit die persepsies van werknemers en bestuur, oorwegend 'n gemeenskaplike eienskap meet en nie onafhanklike konstrukte is soos wel gekonseptualiseer deur deskundiges nie.

  7. Diversity of Vaginal Lactic Acid Bacterial Microbiota in 15 Algerian Pregnant Women with and without Bacterial Vaginosis by using Culture Independent Method.

    Science.gov (United States)

    Alioua, Souad; Abdi, Akila; Fhoula, Imène; Bringel, Françoise; Boudabous, Abdelatif; Ouzari, Imene Hadda

    2016-09-01

    Bacterial Vaginosis (BV) is the most common lower genital tract disorder among women of reproductive age (pregnant and non-pregnant) and a better knowledge of Lactobacillus species richness in healthy and infected vaginal microbiota is needed to efficiently design better probiotic products to promote the maintenance of normal flora which will help prevent bacterial vaginosis. To evaluate and compare the diversity of lactic acid bacterial species in pregnant women with and without BV. A pilot study was carried out during November-2014 to March-2015 in University Badji Mokhtar, Annaba, Algeria. Vaginal swabs were collected from 15 pregnant women aged between 19 and 35 years (mean 27.6 years; n=15) living in the East of Algeria visiting Gynecology service, hospital Abdallah Nouaouria- El bouni, Annaba. Vaginal samples were gram-stained, and scored by the Nugent method. The cohort included cases of women with healthy "normal" vaginal flora, infected flora with bacterial vaginosis and women with "intermediate" flora. The vaginal LAB community from pregnant women was identified by culture independent method based on Denaturing Gradient Gel Electrophoresis (DGGE), with the 16S rRNA gene sequencing. A majority of LAB affiliated to the genus Lactobacillus was found in "normal" and "intermediate" flora (87.5% and 43.75% respectively), while a majority of LAB affiliated to the genus Enterococcus was identified in women with bacterial vaginosis and intermediate flora (60% and 46.75% respectively). Our results showed that the presence of Lactobacillus iners and Lactobacillus delbruekii promotes stability of the vaginal microbiota. This result confirms the findings of previous studies suggesting that the occurrence of predominant Lactobacillus negatively correlates with bacterial vaginosis incidence and their current use as probiotics. Lactobacillus iners and Lactobacillus delbruekii can be defined as critical for defense of the vagina. In addition, Enterococcus feacalis can be

  8. Use of a culture independent method to analyze the diversity of soil fungi surrounding Chroogomphus rutilus in the Beijing region of China

    DEFF Research Database (Denmark)

    Liu, Yu; Wang, Shouxian; Yin, Yonggang

    2012-01-01

    Large-scale cultivation of Chroogomphus rutilus is too inefficient to be commercially feasible. In addition, isolating C. rutilus mycelia in the wild is difficult. Thus, determining the natural habitat of its fruiting body is important. The present study focused on the ecology of the C. rutilus...... habitat to facilitate its large-scale cultivation. A culture-independent molecular approach—a powerful technology for microbiological ecology studies—was used to investigate the diversity of soil fungal communities in samples surrounding C. rutilus from the Beijing region of China. Metagenomic DNA...... was isolated from soil samples collected around C. rutilus, and an internal transcribed spacer (ITS) gene library was constructed. Subsequently, polymerase chain reaction products were digested with HinfI, HaeIII, MspI, TaqI, or MboI. Clones were selected and sequenced based on their restriction fragment...

  9. Culture-independent nested PCR method reveals high diversity of actinobacteria associated with the marine sponges Hymeniacidon perleve and Sponge sp.

    Science.gov (United States)

    Xin, Yanjuan; Huang, Jianyu; Deng, Maicun; Zhang, Wei

    2008-11-01

    A culture-independent nested polymerase chain reaction (PCR) technique was used to investigate the diversity of actinobacteria communities associated with the sponges Hymeniacidon perleve and Sponge sp. The phylogenetic affiliation of sponge-derived actinobacteria was then assessed by 16S rRNA sequencing of cloned DNA fragments. A total of 196 positive clones were screened by restriction fragment length polymorphism (RFLP) analysis; 48 unique operational taxonomic units (OTUs) were selected for sequencing. Rarefaction analysis indicated that the clone libraries represented 93% and 94% of the total estimated diversity for the two species, respectively. Phylogenetic analysis of sequence data revealed representatives of various phylogenetic divisions, which were related to the following ten actinobacterial genera: Acidimicrobium, Corynebacterium, Propionibacterium, Actinomyces, Micrococcus, Microbacterium, Streptomyces, Mycobacterium, Cellulosimicrobium, Sporichthya, and unidentified actinobacterial clones. A sponge-specific, previously uncultured actinobacteria community grouped within the subclass Acidimicrobidae was discovered from both H. perleve and Sponge sp. Sequences belonging to Acidimicrobium in the H. perleve and the Sponge sp. clone libraries represented 33% and 24% of the clones, respectively. In the Sponge sp. clone library Mycobacterium dominated, accounting for 70% of all clones. The presence of Acidimicrobium and mycobacteria within two sponges can lay the groundwork for attempts to culture these interesting bacteria for industrial applications.

  10. Culture-independent discovery of natural products from soil metagenomes.

    Science.gov (United States)

    Katz, Micah; Hover, Bradley M; Brady, Sean F

    2016-03-01

    Bacterial natural products have proven to be invaluable starting points in the development of many currently used therapeutic agents. Unfortunately, traditional culture-based methods for natural product discovery have been deemphasized by pharmaceutical companies due in large part to high rediscovery rates. Culture-independent, or "metagenomic," methods, which rely on the heterologous expression of DNA extracted directly from environmental samples (eDNA), have the potential to provide access to metabolites encoded by a large fraction of the earth's microbial biosynthetic diversity. As soil is both ubiquitous and rich in bacterial diversity, it is an appealing starting point for culture-independent natural product discovery efforts. This review provides an overview of the history of soil metagenome-driven natural product discovery studies and elaborates on the recent development of new tools for sequence-based, high-throughput profiling of environmental samples used in discovering novel natural product biosynthetic gene clusters. We conclude with several examples of these new tools being employed to facilitate the recovery of novel secondary metabolite encoding gene clusters from soil metagenomes and the subsequent heterologous expression of these clusters to produce bioactive small molecules.

  11. Culture Independent Methods for Investigating Microbial Ecology of Fermented Meat%发酵肉制品菌群的非培养鉴定技术研究进展

    Institute of Scientific and Technical Information of China (English)

    王稳航; 徐倩倩; 滕安国; 刘安军

    2014-01-01

    随着分子生物学技术的快速发展,以聚合酶链反应(polymerase chain reaction,PCR)为基础的非培养鉴定技术在发酵肉制品菌群鉴定中得到越来越广泛的应用。其方法主要有种特异性PCR、聚合酶链反应-变性梯度凝胶电泳等。与传统的分离培养鉴定方法相比,非培养方法具有直接、快速、准确、实时等优点,但同时也存在一些缺点,如样品复杂程度、DNA(RNA)提取条件、PCR反应过程的差异等均可影响鉴定结果。非培养鉴定技术与传统微生物鉴定技术相结合,能够对发酵肉制品菌群多态性和动态变化做出准确的鉴定。%With the rapid development of molecular biological technology, culture independent methods based on polymerase chain reaction (PCR) technology have an increasing application in the identification of microorganisms in fermented meat. The existing culture independent methods mainly include species specific PCR and PCR-denatured gradient gel electrophoresis/temperature gradient gel electrophoresis (PCR-DGGE/TGGE). Compared with traditional culture dependent methods, culture independent methods have some advantages such as direct, rapid, accurate and real-time, but also have disadvantages because the results may be influenced by sample complexity, DNA extraction and PCR reaction. The combined application of culture dependent and culture independent methods may provide the best estimation of polymorphism and dynamic changes of microorganisms in fermented meat.

  12. Culture-Independent Molecular Tools for Soil and Rhizosphere Microbiology

    Directory of Open Access Journals (Sweden)

    Peer M. Schenk

    2013-08-01

    Full Text Available Soil microbial communities play an important role in plant health and soil quality. Researchers have developed a wide range of methods for studying the structure, diversity, and activity of microbes to better understand soil biology and plant-microbe interactions. Functional microbiological analyses of the rhizosphere have given new insights into the role of microbial communities in plant nutrition and plant protection against diseases. In this review, we present the most commonly used traditional as well as new culture-independent molecular methods to assess the diversity and function of soil microbial communities. Furthermore, we discuss advantages and disadvantages of these techniques and provide a perspective on emerging technologies for soil microbial community profiling.

  13. Boolean methods of optimization over independence systems

    Energy Technology Data Exchange (ETDEWEB)

    Hulme, B.L.

    1983-01-01

    This paper presents both a direct and an iterative method of solving the combinatorial optimization problem associated with any independence system. The methods use Boolean algebraic computations to produce solutions. In addition, the iterative method employs a version of the greedy algorithm both to compute upper bounds on the optimum value and to produce the additional circuits needed at every stage. The methods are extensions of those used to solve a problem of fire protection at nuclear reactor power plants.

  14. Application of Culture-independent Methods for Analyzing Microbial Communities in Cheese: A Review%非培养技术在干酪微生物群落分析中的应用

    Institute of Scientific and Technical Information of China (English)

    莫蓓红; 孙立国

    2012-01-01

    The application of culture-independent methods in food industry is still in the preliminary stage. In this paper, the application of the most common culture-independent approaches for describing both bacterial and fungal communities in cheese and for monitoring microbial community in cheese manufacturing and ripening process has been reviewed. The most commonly used DNA fingerprinting techniques for investigating microbial communities in cheese are introduced, which include PCR- DGGE, PCR-TTGE, SSCP-PCR, DHPLC and other techniques. Furthermore, the benefits, pitfalls and perspectives of cultureindependent methods in cheese are discussed. Based on the discussion, a polyphasic approach with the combination of culturedependent and culture-independent methods may be the best strategy to achieve more accurate structures of microbial communities.%非培养方法在食品行业中的应用还处于起步阶段。本文综述非培养方法在分析干酪中微生物群落结构以及监测微生物群落结构在干酪生产和成熟过程中演替的应用现状。主要介绍已被广泛用于分析干酪中微生物群落结构的DNA指纹图谱技术,包括PCR.变性梯度凝胶电泳(PCR-DGGE)技术、PCR.时间温度梯度凝胶电泳(PCR-TTGE)技术、单链构象多态性PCR(SSCP-PCR)技术以及变性高效液相色谱(DHPLC)技术等。此外,本文还对非培养技术的优点和局限性进行分析,展望非培养方法在干酪行业的应用前景,认为将非培养方法和传统培养方法结合使用才是研究微生物生态的最好途径。

  15. Comprehensive analysis of secondary dental root canal infections: a combination of culture and culture-independent approaches reveals new insights.

    Directory of Open Access Journals (Sweden)

    Annette Carola Anderson

    Full Text Available Persistence of microorganisms or reinfections are the main reasons for failure of root canal therapy. Very few studies to date have included culture-independent methods to assess the microbiota, including non-cultivable microorganisms. The aim of this study was to combine culture methods with culture-independent cloning methods to analyze the microbial flora of root-filled teeth with periradicular lesions. Twenty-one samples from previously root-filled teeth were collected from patients with periradicular lesions. Microorganisms were cultivated, isolated and biochemically identified. In addition, ribosomal DNA of bacteria, fungi and archaea derived from the same samples was amplified and the PCR products were used to construct clone libraries. DNA of selected clones was sequenced and microbial species were identified, comparing the sequences with public databases. Microorganisms were found in 12 samples with culture-dependent and -independent methods combined. The number of bacterial species ranged from 1 to 12 in one sample. The majority of the 26 taxa belonged to the phylum Firmicutes (14 taxa, followed by Actinobacteria, Proteobacteria and Bacteroidetes. One sample was positive for fungi, and archaea could not be detected. The results obtained with both methods differed. The cloning technique detected several as-yet-uncultivated taxa. Using a combination of both methods 13 taxa were detected that had not been found in root-filled teeth so far. Enterococcus faecalis was only detected in two samples using culture methods. Combining the culture-dependent and -independent approaches revealed new candidate endodontic pathogens and a high diversity of the microbial flora in root-filled teeth with periradicular lesions. Both methods yielded differing results, emphasizing the benefit of combined methods for the detection of the actual microbial diversity in apical periodontitis.

  16. Healing in Herero culture and Namibian African independent churches

    Directory of Open Access Journals (Sweden)

    Selaelo T. Kgatla

    2015-03-01

    Full Text Available The current phenomenon of Namibian African Independent Churches (NAICs draws attention from various people in civil society in Namibia. Although the ministries of NAICs are engaged with activities which are unusual for Christian churches, such as healing the people, fighting against evil spirits and power, performing certain rituals, prophesying and leading the worship services with African Traditional Religion (ATR as a frame of reference in 21st century, they do have a very big influence on various aspects of society in Namibia, which cannot be ignored. This is because those activities are familiar to the everyday lives of Africans and in touch with their culture. With regards to this, this article focuses on the causes of integration or harmony between the Herero culture and the NAICs.

  17. Culture-independent analysis of probiotic products by denaturing gradient gel electrophoresis.

    Science.gov (United States)

    Temmerman, R; Scheirlinck, I; Huys, G; Swings, J

    2003-01-01

    In order to obtain functional and safe probiotic products for human consumption, fast and reliable quality control of these products is crucial. Currently, analysis of most probiotics is still based on culture-dependent methods involving the use of specific isolation media and identification of a limited number of isolates, which makes this approach relatively insensitive, laborious, and time-consuming. In this study, a collection of 10 probiotic products, including four dairy products, one fruit drink, and five freeze-dried products, were subjected to microbial analysis by using a culture-independent approach, and the results were compared with the results of a conventional culture-dependent analysis. The culture-independent approach involved extraction of total bacterial DNA directly from the product, PCR amplification of the V3 region of the 16S ribosomal DNA, and separation of the amplicons on a denaturing gradient gel. Digital capturing and processing of denaturing gradient gel electrophoresis (DGGE) band patterns allowed direct identification of the amplicons at the species level. This whole culture-independent approach can be performed in less than 30 h. Compared with culture-dependent analysis, the DGGE approach was found to have a much higher sensitivity for detection of microbial strains in probiotic products in a fast, reliable, and reproducible manner. Unfortunately, as reported in previous studies in which the culture-dependent approach was used, a rather high percentage of probiotic products suffered from incorrect labeling and yielded low bacterial counts, which may decrease their probiotic potential.

  18. Culture-dependent and -independent investigations of microbial diversity on urinary catheters

    DEFF Research Database (Denmark)

    Xu, Yijuan; Moser, Claus Ernst; Abu Al-Soud, Waleed;

    2012-01-01

    Catheter-associated urinary tract infection is caused by bacteria, which ascend the catheter along its external or internal surface to the bladder and subsequently develop into biofilms on the catheter and uroepithelium. Antibiotic-treated bacteria and bacteria residing in biofilm can be difficul...... to culture. In this study we used culture-based and 16S rRNA gene-based culture-independent methods (fingerprinting, cloning, and pyrosequencing) to determine the microbial diversity of biofilms on 24 urinary catheters. Most of the patients were catheterized for...

  19. Culture-independent microbiological analysis of foley urinary catheter biofilms.

    Directory of Open Access Journals (Sweden)

    Daniel N Frank

    Full Text Available BACKGROUND: Prevention of catheter-associated urinary tract infection (CAUTI, a leading cause of nosocomial disease, is complicated by the propensity of bacteria to form biofilms on indwelling medical devices [1,2,3,4,5]. METHODOLOGY/PRINCIPAL FINDINGS: To better understand the microbial diversity of these communities, we report the results of a culture-independent bacterial survey of Foley urinary catheters obtained from patients following total prostatectomy. Two patient subsets were analyzed, based on treatment or no treatment with systemic fluoroquinolone antibiotics during convalescence. Results indicate the presence of diverse polymicrobial assemblages that were most commonly observed in patients who did not receive systemic antibiotics. The communities typically contained both Gram-positive and Gram-negative microorganisms that included multiple potential pathogens. CONCLUSION/SIGNIFICANCE: Prevention and treatment of CAUTI must take into consideration the possible polymicrobial nature of any particular infection.

  20. Culture-dependent and culture-independent qualitative analysis of probiotic products claimed to contain bifidobacteria.

    Science.gov (United States)

    Masco, L; Huys, G; De Brandt, E; Temmerman, R; Swings, J

    2005-07-15

    A total of 58 probiotic products obtained worldwide, which were claimed to contain Bifidobacterium strains (including 22 yoghurts, 5 dairy fruit drinks, 28 food supplements and 3 pharmaceutical preparations) were investigated in parallel using a culture-dependent and a culture-independent approach. Three isolation media previously reported as selective for Bifidobacterium were evaluated for their suitability in the quality analysis of these products. Subsequently, possible bifidobacterial colonies were picked from the best medium and identified by means of rep-PCR fingerprinting using the BOX primer (BOX-PCR). Bifidobacterium animalis subsp. lactis, formerly classified as Bifidobacterium lactis, was most frequently found, but strains belonging to Bifidobacterium longum biotypes longum and infantis, Bifidobacterium bifidum and Bifidobacterium breve were recovered also. In parallel, all products were also subjected to culture-independent analysis which involved a nested-PCR step on total bacterial DNA extracted directly from the product, followed by separation of the amplicons by Denaturing Gradient Gel Electrophoresis (DGGE) and subsequent identification of species from the band patterns. By conventional cultivation, 70.7% of the products analysed were found to contain culturable bifidobacteria whereas by culture-independent DGGE analysis members of the genus Bifidobacterium could be detected in 96.5% of the analysed products. Genotypic characterization of a number of bifidobacterial isolates at the strain level by means of Pulsed-Field Gel Electrophoresis (PFGE) revealed a relatively high degree of genomic homogeneity among the Bifidobacterium strains currently used in the probiotic industry.

  1. Sampling Natural Viral Communities from Soil for Culture-Independent Analyses

    OpenAIRE

    Williamson, Kurt E.; Wommack, K. Eric; Radosevich, Mark

    2003-01-01

    An essential first step in investigations of viruses in soil is the evaluation of viral recovery methods suitable for subsequent culture-independent analyses. In this study, four elution buffers (10% beef extract, 250 mM glycine buffer, 10 mM sodium pyrophosphate, and 1% potassium citrate) and three enumeration techniques (plaque assay, epifluorescence microscopy [EFM], and transmission electron microscopy [TEM]) were compared to determine the best method of extracting autochthonous bacteriop...

  2. Metagenomics: The Next Culture-Independent Game Changer

    Directory of Open Access Journals (Sweden)

    Jessica D. Forbes

    2017-07-01

    Full Text Available A trend towards the abandonment of obtaining pure culture isolates in frontline laboratories is at a crossroads with the ability of public health agencies to perform their basic mandate of foodborne disease surveillance and response. The implementation of culture-independent diagnostic tests (CIDTs including nucleic acid and antigen-based assays for acute gastroenteritis is leaving public health agencies without laboratory evidence to link clinical cases to each other and to food or environmental substances. This limits the efficacy of public health epidemiology and surveillance as well as outbreak detection and investigation. Foodborne outbreaks have the potential to remain undetected or have insufficient evidence to support source attribution and may inadvertently increase the incidence of foodborne diseases. Next-generation sequencing of pure culture isolates in clinical microbiology laboratories has the potential to revolutionize the fields of food safety and public health. Metagenomics and other ‘omics’ disciplines could provide the solution to a cultureless future in clinical microbiology, food safety and public health. Data mining of information obtained from metagenomics assays can be particularly useful for the identification of clinical causative agents or foodborne contamination, detection of AMR and/or virulence factors, in addition to providing high-resolution subtyping data. Thus, metagenomics assays may provide a universal test for clinical diagnostics, foodborne pathogen detection, subtyping and investigation. This information has the potential to reform the field of enteric disease diagnostics and surveillance and also infectious diseases as a whole. The aim of this review will be to present the current state of CIDTs in diagnostic and public health laboratories as they relate to foodborne illness and food safety. Moreover, we will also discuss the diagnostic and subtyping utility and concomitant bias limitations of

  3. Metagenomics: The Next Culture-Independent Game Changer

    Science.gov (United States)

    Forbes, Jessica D.; Knox, Natalie C.; Ronholm, Jennifer; Pagotto, Franco; Reimer, Aleisha

    2017-01-01

    A trend towards the abandonment of obtaining pure culture isolates in frontline laboratories is at a crossroads with the ability of public health agencies to perform their basic mandate of foodborne disease surveillance and response. The implementation of culture-independent diagnostic tests (CIDTs) including nucleic acid and antigen-based assays for acute gastroenteritis is leaving public health agencies without laboratory evidence to link clinical cases to each other and to food or environmental substances. This limits the efficacy of public health epidemiology and surveillance as well as outbreak detection and investigation. Foodborne outbreaks have the potential to remain undetected or have insufficient evidence to support source attribution and may inadvertently increase the incidence of foodborne diseases. Next-generation sequencing of pure culture isolates in clinical microbiology laboratories has the potential to revolutionize the fields of food safety and public health. Metagenomics and other ‘omics’ disciplines could provide the solution to a cultureless future in clinical microbiology, food safety and public health. Data mining of information obtained from metagenomics assays can be particularly useful for the identification of clinical causative agents or foodborne contamination, detection of AMR and/or virulence factors, in addition to providing high-resolution subtyping data. Thus, metagenomics assays may provide a universal test for clinical diagnostics, foodborne pathogen detection, subtyping and investigation. This information has the potential to reform the field of enteric disease diagnostics and surveillance and also infectious diseases as a whole. The aim of this review will be to present the current state of CIDTs in diagnostic and public health laboratories as they relate to foodborne illness and food safety. Moreover, we will also discuss the diagnostic and subtyping utility and concomitant bias limitations of metagenomics and comparable

  4. Biodiversity in Oscypek, a traditional Polish cheese, determined by culture-dependent and -independent approaches.

    Science.gov (United States)

    Alegría, Angel; Szczesny, Pawel; Mayo, Baltasar; Bardowski, Jacek; Kowalczyk, Magdalena

    2012-03-01

    Oscypek is a traditional Polish scalded-smoked cheese, with a protected-designation-of-origin (PDO) status, manufactured from raw sheep's milk without starter cultures in the Tatra Mountains region of Poland. This study was undertaken in order to gain insight into the microbiota that develops and evolves during the manufacture and ripening stages of Oscypek. To this end, we made use of both culturing and the culture-independent methods of PCR followed by denaturing gradient gel electrophoresis (PCR-DGGE) and pyrosequencing of 16S rRNA gene amplicons. The culture-dependent technique and PCR-DGGE fingerprinting detected the predominant microorganisms in traditional Oscypek, whereas the next-generation sequencing technique (454 pyrosequencing) revealed greater bacterial diversity. Besides members of the most abundant bacterial genera in dairy products, e.g., Lactococcus, Lactobacillus, Leuconostoc, Streptococcus, and Enterococcus, identified by all three methods, other, subdominant bacteria belonging to the families Bifidobacteriaceae and Moraxellaceae (mostly Enhydrobacter), as well as various minor bacteria, were identified by pyrosequencing. The presence of bifidobacterial sequences in a cheese system is reported for the first time. In addition to bacteria, a great diversity of yeast species was demonstrated in Oscypek by the PCR-DGGE method. Culturing methods enabled the determination of a number of viable microorganisms from different microbial groups and their isolation for potential future applications in specific cheese starter cultures.

  5. Culture-independent analysis of bacterial diversity in a child-care facility

    Directory of Open Access Journals (Sweden)

    Tin Sara

    2007-04-01

    Full Text Available Abstract Background Child-care facilities appear to provide daily opportunities for exposure and transmission of bacteria and viruses. However, almost nothing is known about the diversity of microbial contamination in daycare facilities or its public health implications. Recent culture-independent molecular studies of bacterial diversity in indoor environments have revealed an astonishing diversity of microorganisms, including opportunistic pathogens and many uncultured bacteria. In this study, we used culture and culture-independent methods to determine the viability and diversity of bacteria in a child-care center over a six-month period. Results We sampled surface contamination on toys and furniture using sterile cotton swabs in four daycare classrooms. Bacteria were isolated on nutrient and blood agar plates, and 16S rRNA gene sequences were obtained from unique (one of a kind colony morphologies for species identification. We also extracted DNA directly from nine representative swab samples taken over the course of the study from both toy and furniture surfaces, and used "universal" 16S rRNA gene bacterial primers to create PCR-based clone libraries. The rRNA gene clones were sequenced, and the sequences were compared with related sequences in GenBank and subjected to phylogenetic analyses to determine their evolutionary relationships. Culturing methods identified viable bacteria on all toys and furniture surfaces sampled in the study. Bacillus spp. were the most commonly cultured bacteria, followed by Staphylococcus spp., and Microbacterium spp. Culture-independent methods based on 16S rRNA gene sequencing, on the other hand, revealed an entirely new dimension of microbial diversity, including an estimated 190 bacterial species from 15 bacterial divisions. Sequence comparisons and phylogenetic analyses determined that the clone libraries were dominated by a diverse set of sequences related to Pseudomonas spp., as well as uncultured

  6. Analysis of the diversity of water kefir microbiota by culture-dependent and -independent approaches

    OpenAIRE

    Gulitz, Anna Jana

    2014-01-01

    Water kefir is a traditional fermented beverage based on a sucrose solution with different dried fruits. In this work the microbiome of water kefir and the dynamic of the water kefir consortium depending on the cultivation conditions was analyzed by culture-dependent and –independent methods. Microorganisms important for the water kefir grain formation and for the typical beverage characteristics were identified. The most abundant species were Lactobacillus nagelii, Lb. hordei, Leuconostoc me...

  7. Mesh-Independent Methods for Agent Movement

    Science.gov (United States)

    2006-02-01

    effectively in ecology , where it is often called individual based modeling (IBM)[1]. Some individual based models are spatially explicit meaning that...3-D movement rules of fish for forecasting using a coupled Eulerian- Lagrangian-Agent framework. Journal of Ecological Modeling, 2005. in press. [2...refinement using parallel in-element particle track- ing methods. In José M.L.M. Palma , Jack Dongarra, Vicente Hernández, REFERENCES 21 and A

  8. Comparison of culture-dependent and independent approaches to characterize fecal bifidobacteria and lactobacilli.

    Science.gov (United States)

    Quartieri, Andrea; Simone, Marta; Gozzoli, Caterina; Popovic, Mina; D'Auria, Giuseppe; Amaretti, Alberto; Raimondi, Stefano; Rossi, Maddalena

    2016-04-01

    Different culture-dependent and independent methods were applied to investigate the population of bifidobacteria and lactobacilli in the feces of five healthy subjects. Bacteria were isolated on MRS, a complex medium supporting growth of lactobacilli and bifidobacteria, and on three selective media for bifidobacteria and two for lactobacilli. Taxonomic characterization of the isolates was carried out by RAPD-PCR and partial 16S sequencing. The selectivity of genus-specific media was also investigated by challenging colonies from MRS plates to grow onto each medium. In parallel, a quantitative and qualitative description of bifidobacteria and lactic acid bacteria was obtained by FISH, qPCR, TRFLP, and 16S rRNA gene sequencing. Bifidobacteria did not fail to grow on their specific media and were easily isolated and enumerated, showing comparable quantitative data among culture-dependent and -independent techniques. The Bifidobacterium species identified on plates and those extracted from TRFLP and 16S rRNA gene sequencing were mostly overlapping. Selective media for lactobacilli gave unsuitable results, being too stringent or too permissive. The quantification of lactobacilli through selective plates, qPCR, FISH, and 16S rRNA gene sequencing gave unreliable results. Therefore, unlike bifidobacteria, intestinal lactobacilli are still problematic in terms of quantification and accurate profiling at level of species and possibly of strains by both culture-dependent and culture-independent techniques.

  9. Culture-Independent Identification of Nontuberculous Mycobacteria in Cystic Fibrosis Respiratory Samples.

    Directory of Open Access Journals (Sweden)

    Lindsay J Caverly

    Full Text Available Respiratory tract infections with nontuberculous mycobacteria (NTM are increasing in prevalence and are a significant cause of lung function decline in individuals with cystic fibrosis (CF. NTM have been detected in culture-independent analyses of CF airway microbiota at lower rates than would be expected based on published prevalence data, likely due to poor lysing of the NTM cell wall during DNA extraction. We compared a standard bacterial lysis protocol with a modified method by measuring NTM DNA extraction by qPCR and NTM detection with bacterial 16S rRNA gene sequencing. The modified method improved NTM DNA recovery from spiked CF sputum samples by a mean of 0.53 log10 copies/mL for M. abscessus complex and by a mean of 0.43 log10 copies/mL for M. avium complex as measured by qPCR targeting the atpE gene. The modified method also improved DNA sequence based NTM detection in NTM culture-positive CF sputum and bronchoalveolar lavage samples; however, both qPCR and 16S rRNA gene sequencing remained less sensitive than culture for NTM detection. We highlight the limitations of culture-independent identification of NTM from CF respiratory samples, and illustrate how alterations in the bacterial lysis and DNA extraction process can be employed to improve NTM detection with both qPCR and 16S rRNA gene sequencing.

  10. Experimental implementation of the modified independent modal space control method

    Science.gov (United States)

    Baz, A.; Poh, S.

    1990-01-01

    An experimental realization of a modified independent modal space control (MIMSC) method to control the vibration of a flexible cantilevered beam is presented. In its operation the method relies on the use of one piezoelectric actuator to control several vibration modes through a time-sharing strategy. The effectiveness of the MIMSC method in damping out the beam vibration is demonstrated by comparing the results with those obtained by other modal control methods. Two methods are considered, the independent modal space control method and the pseudo-inverse method. The feasibility of the MIMSC method as a viable alternative for controlling large flexible structures with a very small number of actuators is emphasized.

  11. Culture-independent qunatification of Salmonella enterica in carcass gauze swabs by flotation prior to real-time PCR

    DEFF Research Database (Denmark)

    Löfström, Charlotta; Schelin, Jenny; Norling, Börje

    2010-01-01

    To facilitate quantitative risk assessment in the meat production chain, there is a need for culture-independent quantification methods. The aim of this study was to evaluate the use of flotation, a non-destructive sample preparation method based on traditional buoyant density centrifugation, for...

  12. Making Mavericks: Preparing Musicians for Independent Artistic Culture

    Science.gov (United States)

    Canham, Nicole L.

    2016-01-01

    This essay explores the role that maverick qualities--"independent or unorthodox behaviour" ("The Oxford Dictionary," 2015)--play in developing and sustaining musician employability. Whilst career education for musicians often highlights new career models (Bridgstock, 2005), there is limited evidence of how these concepts work…

  13. Biodeterioration of epoxy resin: a microbial survey through culture-independent and culture-dependent approaches.

    Science.gov (United States)

    Pangallo, Domenico; Bučková, Maria; Kraková, Lucia; Puškárová, Andrea; Šaková, Nikoleta; Grivalský, Tomaš; Chovanová, Katarina; Zemánková, Milina

    2015-02-01

    During the 20th century, synthetic polymers were greatly used in the field of art. In particular, the epoxy resins were used for both conservation and for creating sculptures. The biodeterioration of these polymers has not been adequately studied. The aim of this investigation was to examine the microflora responsible for the deterioration of an epoxy statue exposed to outdoor conditions. Fungal and bacterial microflora were isolated from the art object, clustered by fluorescence-ITS (internal transcribed spacer), identified by ITS and 16S rRNA sequencing and tested for their lipolytic abilities by three agar assays. Different algal, bacterial, cyanobacterial and fungal clone libraries were constructed. The surrounding airborne microflora was analyzed using culture-dependent and culture-independent approaches. The results indicated the presence, on the statue surface, of an interesting and differentiate microbial community composed of rock-inhabiting members, algal photobionts (Trebouxia spp., Chloroidium ellipsoideum and Chlorella angustoellipsoidea), Cyanobacteria (Leptolyngbya sp., Phormidium sp., Cylindrospermum stagnale, Hassallia byssoidea and Geitlerinema sp.), black yeasts related to the species Friedmanniomyces endolithicus, Pseudotaeniolina globosa, Phaeococcomyces catenatus and Catenulostroma germanicum and several plant-associated fungi. This investigation provides new information on the potential microfloral inhabitants of epoxy resin discovering a new ecological niche, occupied mainly by several members of rock-colonizing microbial species.

  14. Culture-independent quantification of Salmonella enterica in carcass gauze swabs by flotation prior to real-time PCR

    DEFF Research Database (Denmark)

    Löfström, Charlotta; Schelin, Jenny; Norling, Börje

    2011-01-01

    To facilitate quantitative risk assessment in the meat production chain, there is a need for culture-independent quantification methods. The aim of this study was to evaluate the use of flotation, a non-destructive sample preparation method based on traditional buoyant density centrifugation......, for culture-independent quantification of intact Salmonella in pig carcass gauze swabs (100 cm2) prior to quantitative PCR (qPCR). A novel approach was investigated, excluding the homogenization step prior to flotation, to improve the detection limit and speed up the quantification procedure. The buoyant...... concentrations of ≥ 6.1 × 108 CFU/swab sample, but not by concentrations ≤ 6.1 × 106 CFU/swab sample. By using the gauze swabs directly in the flotation procedure, the homogenization step normally used for preparation of food-related samples could be excluded, which simplified the culture independent...

  15. Culture-independent qunatification of Salmonella enterica in carcass gauze swabs by flotation prior to real-time PCR

    DEFF Research Database (Denmark)

    Löfström, Charlotta; Schelin, Jenny; Norling, Börje

    2010-01-01

    To facilitate quantitative risk assessment in the meat production chain, there is a need for culture-independent quantification methods. The aim of this study was to evaluate the use of flotation, a non-destructive sample preparation method based on traditional buoyant density centrifugation......, for culture-independent quantification of intact Salmonella in pig carcass gauze swabs (100 cm2) prior to quantitative PCR (qPCR). A novel approach was investigated, excluding the homogenization step prior to flotation, to improve the detection limit and speed up the quantification procedure. The buoyant...... concentrations of ≥ 6.1×108 CFU/swab sample, but not by concentrations ≤ 6.1×106 CFU/swab sample. By using the gauze swabs directly in the flotation procedure, the homogenization step normally used for preparation of food-related samples could be excluded, which simplified the culture independent quantification...

  16. Culture-independent quantification of Salmonella enterica in carcass gauze swabs by flotation prior to real-time PCR

    DEFF Research Database (Denmark)

    Löfström, Charlotta; Schelin, Jenny; Norling, Börje

    2011-01-01

    To facilitate quantitative risk assessment in the meat production chain, there is a need for culture-independent quantification methods. The aim of this study was to evaluate the use of flotation, a non-destructive sample preparation method based on traditional buoyant density centrifugation......, for culture-independent quantification of intact Salmonella in pig carcass gauze swabs (100 cm2) prior to quantitative PCR (qPCR). A novel approach was investigated, excluding the homogenization step prior to flotation, to improve the detection limit and speed up the quantification procedure. The buoyant...... concentrations of ≥ 6.1 × 108 CFU/swab sample, but not by concentrations ≤ 6.1 × 106 CFU/swab sample. By using the gauze swabs directly in the flotation procedure, the homogenization step normally used for preparation of food-related samples could be excluded, which simplified the culture independent...

  17. Bibliography of Methods in Cultural Anthropology.

    Science.gov (United States)

    Dow, James, Comp.

    Over 300 resources on methods in cultural anthropology are listed under the following headings: archaeological methods; visual methods, tape recordings, and technical aids; cognitive anthropology and emic methods; community studies and complex societies; cross-cultural and hologeistic methods; ethnohistory; field work techniques and participant…

  18. The Method of Culture Contrast

    DEFF Research Database (Denmark)

    Hasse, Cathrine; Trentemøller, Stine; Motzkau, Johanna

    2009-01-01

    the researcher's cultural background (i.e., emic categorisations) in the analysis of cross-cultural comparisons we suggest that the categorisations the researcher use in her tests and fieldwork descriptions are taken to be part of the research itself, rather than simply being an underlying (taken for granted......In this article, we suggest that research is a practical activity building on local category systems belonging specifically to research (etic categories) as well as categories belonging specifically to the national culture of the researcher (emic categories) (Pike 1967). Much cross-cultural......) framework on which the research is conducted. First we present a recent study of European universities as culturally diverse working places and we present an approach in which the researcher's emic and etic categorisations can be challenged when contrasted with each other (Hasse & Trentemøller 2008). Second...

  19. Independência dos auditores portugueses : o efeito cultural

    OpenAIRE

    Pinheiro, Bruno Miguel Braia

    2013-01-01

    Mestrado em Contabilidade, Fiscalidade e Finanças Empresariais Este estudo pretendeu analisar a importância dos factores que afectam a independência dos auditores (ou seja, as necessidades do investidor, retenção de clientes, valores e obrigações profissionais e pressões de tempo/orçamentárias), em dois cenários e em fase de realização dos trabalhos de auditoria. O primeiro cenário (caso 1) está relacionado com a pressão de tempo em face do prazo de entrega do relatório de auditoria, enqua...

  20. Halo-independent methods for inelastic dark matter scattering

    CERN Document Server

    Bozorgnia, Nassim; Schwetz, Thomas; Zupan, Jure

    2013-01-01

    We present halo-independent methods to analyze the results of dark matter direct detection experiments assuming inelastic scattering. We focus on the annual modulation signal reported by DAMA/LIBRA and present three different halo-independent tests. First, we compare it to the upper limit on the unmodulated rate from XENON100 using (a) the trivial requirement that the amplitude of the annual modulation has to be smaller than the bound on the unmodulated rate, and (b) a bound on the annual modulation amplitude based on an expansion in the Earth's velocity. The third test uses the special predictions of the signal shape for inelastic scattering and allows for an internal consistency check of the data without referring to any astrophysics. We conclude that a strong conflict between DAMA/LIBRA and XENON100 in the framework of spin-independent inelastic scattering can be established independently of the local properties of the dark matter halo.

  1. Culture Independent Geochemical Tools for Adressing Microbial Activity

    Science.gov (United States)

    Lomstein, B. A.; Langerhuus, A. T.; Jørgensen, B. B.; Alperin, M. J.

    2014-12-01

    Decades of ocean drilling have demonstrated wide-spread microbial life in deep sub-seafloor sediment, and surprisingly high numbers of microbial cells and endospores. Despite the ubiquity of life in the deep biosphere, the large community sizes are not yet understood given the extremely low energy fluxes. We have developed and applied new approaches to the deep sub-seafloor to quantify distributions and turnover times of living microbial biomass, endospores and microbial necromass. The approach combines sensitive analyses of unique bacterial marker molecules (muramic acid and d-amino acids) and the bacterial endospore marker (dipicolinic acid) with a series of models that link microscopic (e.g., racemization dynamics of stereo-isomeric amino acids) and macroscopic (e.g., porewater geochemistry) properties. Model output includes production rates and turnover times of microbial biomass and necromass, concentration profiles of reactive organic carbon, and rates of organic carbon decomposition. In combination, these results allow us to assess the role of microbial activity in the sub-seafloor carbon budget. One key result is that the turnover time of biomass is far longer than turnover times found in cultures and active surface sediments.

  2. Theory and methods in cultural neuroscience

    Science.gov (United States)

    Hariri, Ahmad R.; Harada, Tokiko; Mano, Yoko; Sadato, Norihiro; Parrish, Todd B.; Iidaka, Tetsuya

    2010-01-01

    Cultural neuroscience is an emerging research discipline that investigates cultural variation in psychological, neural and genomic processes as a means of articulating the bidirectional relationship of these processes and their emergent properties. Research in cultural neuroscience integrates theory and methods from anthropology, cultural psychology, neuroscience and neurogenetics. Here, we review a set of core theoretical and methodological challenges facing researchers when planning and conducting cultural neuroscience studies, and provide suggestions for overcoming these challenges. In particular, we focus on the problems of defining culture and culturally appropriate experimental tasks, comparing neuroimaging data acquired from different populations and scanner sites and identifying functional genetic polymorphisms relevant to culture. Implications of cultural neuroscience research for addressing current issues in population health disparities are discussed. PMID:20592044

  3. Evidence for a cultural influence on field-independence in autism spectrum disorder.

    Science.gov (United States)

    Koh, Hwan Cui; Milne, Elizabeth

    2012-02-01

    Field-independence, or weak central coherence, is a recognised phenotype of autism spectrum disorder (ASD). There is also evidence of cultural variation in this perceptual style, as neurotypical individuals from Western nations are more field-independent than neurotypical individuals from East-Asian nations. The majority of research on perceptual style in those with ASD has been carried out in Western nations therefore it is unclear whether increased field-independence in ASD is a culturally universal phenotype. Here, we assessed perceptual style in children with and without ASD from England and Singapore using the Children's Embedded Figures Test and the Framed-Line Test. We found increased field-independence in the English participants with ASD only, suggesting that weak central coherence in ASD is not culturally universal.

  4. Action Methods for Teaching Cultural Diversity Awareness.

    Science.gov (United States)

    Tomasulo, Dan

    This paper is devoted to the description of action methods that can be used to provide a practical understanding and awareness of culturally diverse material. It draws from such varied disciplines as cross-cultural psychology, international business, and sociodrama, with the goal of suggesting a methodology for using role playing to teach ethnic,…

  5. Characterization of progesterone-independent avidin production of chicken tissues in culture.

    Science.gov (United States)

    Nordback, I; Kulomaa, M; Joensuu, T; Tuohimaa, P

    1982-01-01

    1. Primary cell cultures of chick oviduct, intestine, peritoneal membrane and lung, but not those of wing muscle or calcaneal tendon, produced avidin progesterone-independently through a 5-week culture. 2. Actinomycin D and cycloheximide, but not hydroxyurea, inhibited the avidin production. 3. The common feature of the cultures producing avidin was a fibroblast-like cell type. 4. Similarly, oviduct and lung, but not muscle or tendon, also produced avidin in hormone-free short-term organ cultures, where the avidin synthesis began during the first 8 h of culture. 5. Induction of the general protein synthesis does not explain the avidin production, since another egg-white protein, ovalbumin, was not induced by the culture procedure.

  6. Culture and Healthy Eating: The Role of Independence and Interdependence in the United States and Japan.

    Science.gov (United States)

    Levine, Cynthia S; Miyamoto, Yuri; Markus, Hazel Rose; Rigotti, Attilio; Boylan, Jennifer Morozink; Park, Jiyoung; Kitayama, Shinobu; Karasawa, Mayumi; Kawakami, Norito; Coe, Christopher L; Love, Gayle D; Ryff, Carol D

    2016-10-01

    Healthy eating is important for physical health. Using large probability samples of middle-aged adults in the United States and Japan, we show that fitting with the culturally normative way of being predicts healthy eating. In the United States, a culture that prioritizes and emphasizes independence, being independent predicts eating a healthy diet (an index of fish, protein, fruit, vegetables, reverse-coded sugared beverages, and reverse-coded high fat meat consumption; Study 1) and not using nonmeat food as a way to cope with stress (Study 2a). In Japan, a culture that prioritizes and emphasizes interdependence, being interdependent predicts eating a healthy diet (Studies 1 and 2b). Furthermore, reflecting the types of agency that are prevalent in each context, these relationships are mediated by autonomy in the United States and positive relations with others in Japan. These findings highlight the importance of understanding cultural differences in shaping healthy behavior and have implications for designing health-promoting interventions.

  7. Development and evaluation of a culture-independent method for source determination of fecal wastes in surface and storm waters using reverse transcriptase-PCR detection of FRNA coliphage genogroup gene sequences.

    Science.gov (United States)

    A complete method, incorporating recently improved reverse transcriptase-PCR primer/probe assays and including controls for determining interferences to phage recoveries from water sample concentrates and for detecting interferences to their analysis, was developed for the direct...

  8. Development and evaluation of a culture-independent method for source determination of fecal wastes in surface and storm waters using reverse transcriptase-PCR detection of FRNA coliphage genogroup gene sequences.

    Science.gov (United States)

    A complete method, incorporating recently improved reverse transcriptase-PCR primer/probe assays and including controls for determining interferences to phage recoveries from water sample concentrates and for detecting interferences to their analysis, was developed for the direct...

  9. Fungal diversity from various marine habitats deduced through culture-independent studies

    Digital Repository Service at National Institute of Oceanography (India)

    Manohar, C.S.; Raghukumar, C.

    Author version: FEMS Microbiol. Lett., vol.341; 2013; 69-78 Fungal diversity from various marine habitats deduced through culture-independent studies Cathrine Sumathi Manohar* and Chandralata Raghukumar$ National Institute of Oceanography, (Council...: un-cultured fungal diversity from marine habitats Abstract Studies on the molecular diversity of the micro-eukaryotic community have shown that fungi occupy a central position in a large number of marine habitats. Environmental surveys using...

  10. The frequency-independent control method for distributed generation systems

    DEFF Research Database (Denmark)

    Naderi, Siamak; Pouresmaeil, Edris; Gao, Wenzhong David

    2012-01-01

    In this paper a novel frequency-independent control method suitable for distributed generation (DG) is presented. This strategy is derived based on the . abc/. αβ transformation and . abc/. dq transformation of the ac system variables. The active and reactive currents injected by the DG...... are controlled in the synchronously rotating orthogonal . dq reference frame. The transformed variables are used in control of the voltage source inverter that connects DG to distribution network. Due to importance of distributed resources in modern power systems, development of new, practical, cost...

  11. Why are price stability and statutory independence of central banks negatively correlated? : the role of culture

    NARCIS (Netherlands)

    Jong, Eelke de

    2001-01-01

    This paper investigates whether in OECD-countries the negative relation between central bank independence and inflation is related to culture, in the sense of common values and norms. It appears that inflation is lower in countries where people dislike uncertainty. The tolerance in a society with

  12. 基于免培养技术的食用菌病害微生物rDNA测序及初步分析%rDNA Sequencing and Analysis of Edible Fungus Disease Microorganisms Using Culture-Independent Method

    Institute of Scientific and Technical Information of China (English)

    高能; 姚强; 宫志远; 郭立忠; 万鲁长; 任鹏飞; 韩建东; 黄春燕

    2013-01-01

    The microorganism metagenome of four kinds of edible fungus diseases was extracted by CTAB method combined with DNA gel recovery kit.The genomic DNA was amplified respectively by polymerase chain reaction using the universal primers of the 16S rDNA and 18S rDNA gene,and then monoclonal sequenced after hgated and transformed.At least 20 positive clones selected by random of each pair of primers were performed sequence alignment.The results showed that there were two bacterial diseases and two fungal diseases in the samples.%采用CTAB结合DNA凝胶回收试剂盒法提取四种主要食用菌病害组织样品的宏基因组DNA,用16S rDNA和18S rDNA通用引物分别对样品的基因组DNA进行PCR扩增、连接、转化并进行单克隆测序.每对引物随机挑取20个阳性克隆的rDNA序列进行比对,并对病害微生物的亲缘关系进行了初步分析,结果表明四种病害样品中两种为细菌性病害,两种为真菌性病害.

  13. Cultural orientations, parental beliefs and practices, and latino adolescents' autonomy and independence.

    Science.gov (United States)

    Roche, Kathleen M; Caughy, Margaret O; Schuster, Mark A; Bogart, Laura M; Dittus, Patricia J; Franzini, Luisa

    2014-08-01

    Despite the salience of behavioral autonomy and independence to parent-child interactions during middle adolescence, little is known about parenting processes pertinent to youth autonomy development for Latino families. Among a diverse sample of 684 Latino-origin parent-adolescent dyads in Houston, Texas, this study examines how parents' cultural orientations are associated directly and indirectly, through parental beliefs, with parenting practices giving youth behavioral autonomy and independence. Informed by social domain theory, the study's parenting constructs pertain to youth behaviors in an "ambiguously personal" domain-activities that adolescents believe are up to youth to decide, but which parents might argue require parents' supervision, knowledge, and/or decision-making. Results for latent profile analyses of parents' cultural identity across various facets of acculturation indicate considerable cultural heterogeneity among Latino parents. Although 43% of parents have a Latino cultural orientation, others represent Spanish-speaking/bicultural (21%), bilingual/bicultural (15%), English-speaking/bicultural (15%), or US (6%) cultural orientations. Structural equation modeling results indicate that bilingual/bicultural, English-speaking/bicultural, and US-oriented parents report less emphasis on the legitimacy of parental authority and younger age expectations for youth to engage in independent behaviors than do Latino-oriented parents. Parental beliefs endorsing youth's behavioral independence and autonomy, in turn, are associated with less stringent parental rules (parental report), less parental supervision (parental and youth report), and more youth autonomy in decision-making (parental and youth report). Evidence thus supports the idea that the diverse cultural orientations of Latino parents in the US may result in considerable variations in parenting processes pertinent to Latino adolescents' development.

  14. Culture-independent detection of microorganisms in traditional Slovakian bryndza cheese.

    Science.gov (United States)

    Chebeňová-Turcovská, Viera; Zenišová, Katarína; Kuchta, Tomáš; Pangallo, Domenico; Brežná, Barbara

    2011-10-17

    In order to investigate the microflora of Slovakian bryndza cheese (a cheese containing unpasteurized or pasteurized ewes' milk component) by a culture-independent method, DNA was extracted directly from 7 bryndza samples and analysed by an innovative method. Using the universal prokaryotic and fungal primers, ribosomal DNA internal transcribed spacer (ITS) regions with variable length were amplified. The standard universal reverse primer L1 aligning to bacterial 23s rDNA was found unsuitable for some lactic acid bacteria and other species based on in silico analysis. Therefore, L1 primer was replaced by a combination of novel primers GplusR and GminusR aligning to the adjacent, more conserved DNA region. The amplification profiles were visualised by both standard electrophoresis and by fluorescent capillary gel electrophoresis. From representative samples, major amplicons were excised from the gel, cloned and sequenced. Sequencing revealed that the samples contained Lactobacillus delbrueckii, Lactobacillus brevis, Streptococcus thermophilus, Lactococcus lactis, Lactococcus raffinolactis, Streptococcus macedonicus, Leuconostoc pseudomesenteroides, Debaromyces hansenii, Mucor fragilis, Yarrowia lipolytica and Galactomyces geotrichum. These results represent an extension of the knowledge on the microflora of Slovakian bryndza cheese. The introduced automated ribosomal DNA intergenic spacer analysis of the bacterial and fungal genomes proved to be very effective in the application of studying microflora of cheese.

  15. Acetic acid bacteria from biofilm of strawberry vinegar visualized by microscopy and detected by complementing culture-dependent and culture-independent techniques.

    Science.gov (United States)

    Valera, Maria José; Torija, Maria Jesús; Mas, Albert; Mateo, Estibaliz

    2015-04-01

    Acetic acid bacteria (AAB) usually develop biofilm on the air-liquid interface of the vinegar elaborated by traditional method. This is the first study in which the AAB microbiota present in a biofilm of vinegar obtained by traditional method was detected by pyrosequencing. Direct genomic DNA extraction from biofilm was set up to obtain suitable quality of DNA to apply in culture-independent molecular techniques. The set of primers and TaqMan--MGB probe designed in this study to enumerate the total AAB population by Real Time--PCR detected between 8 × 10(5) and 1.2 × 10(6) cells/g in the biofilm. Pyrosequencing approach reached up to 10 AAB genera identification. The combination of culture-dependent and culture-independent molecular techniques provided a broader view of AAB microbiota from the strawberry biofilm, which was dominated by Ameyamaea, Gluconacetobacter, and Komagataeibacter genera. Culture-dependent techniques allowed isolating only one genotype, which was assigned into the Ameyamaea genus and which required more analysis for a correct species identification. Furthermore, biofilm visualization by laser confocal microscope and scanning electronic microscope showed different dispositions and cell morphologies in the strawberry vinegar biofilm compared with a grape vinegar biofilm.

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

    Directory of Open Access Journals (Sweden)

    R. Urbano

    2011-02-01

    Full Text Available 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.

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

    Science.gov (United States)

    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.

  18. [Culture based diagnostic methods for tuberculosis].

    Science.gov (United States)

    Baylan, Orhan

    2005-01-01

    Culture methods providing isolates for identification and drug susceptibility testing, still represent the gold standard for the definitive diagnosis of tuberculosis, although the delay in obtaining results still remains a problem. Traditional solid media are recommended for use along with liquid media in primary isolation of mycobacteria. At present, a number of elaborate culture systems are available commercially. They range from simple bottles and tubes such as MGIT (BD Diagnostic Systems, USA), Septi-Chek AFB (BD, USA) and MB Redox (Biotest Diagnostics, USA) to semiautomated system (BACTEC 460TB, BD, USA) and fully automated systems (BACTEC 9000 MB [BD, USA], BACTEC MGIT 960 [BD, USA], ESP Culture System II [Trek Diagnostics, USA], MB/BacT ALERT 3D System [BioMérieux, NC], TK Culture System [Salubris Inc, Turkey]). Culture methods available today are sufficient to permit laboratories to develop an algoritm that is optimal for patients and administrative needs. In this review article, the culture systems used for the diagnosis of tuberculosis, their mechanisms, advantages and disadvantages have been discussed under the light of recent literature.

  19. Culture-independent investigation of the microbiome associated with the nematode Acrobeloides maximus.

    Directory of Open Access Journals (Sweden)

    Jean-Paul Baquiran

    Full Text Available BACKGROUND: Symbioses between metazoans and microbes are widespread and vital to many ecosystems. Recent work with several nematode species has suggested that strong associations with microbial symbionts may also be common among members of this phylu. In this work we explore possible symbiosis between bacteria and the free living soil bacteriovorous nematode Acrobeloides maximus. METHODOLOGY: We used a soil microcosm approach to expose A. maximus populations grown monoxenically on RFP labeled Escherichia coli in a soil slurry. Worms were recovered by density gradient separation and examined using both culture-independent and isolation methods. A 16S rRNA gene survey of the worm-associated bacteria was compared to the soil and to a similar analysis using Caenorhabditis elegans N2. Recovered A. maximus populations were maintained on cholesterol agar and sampled to examine the population dynamics of the microbiome. RESULTS: A consistent core microbiome was extracted from A. maximus that differed from those in the bulk soil or the C. elegans associated set. Three genera, Ochrobactrum, Pedobacter, and Chitinophaga, were identified at high levels only in the A. maximus populations, which were less diverse than the assemblage associated with C. elegans. Putative symbiont populations were maintained for at least 4 months post inoculation, although the levels decreased as the culture aged. Fluorescence in situ hybridization (FISH using probes specific for Ochrobactrum and Pedobacter stained bacterial cells in formaldehyde fixed nematode guts. CONCLUSIONS: Three microorganisms were repeatedly observed in association with Acrobeloides maximus when recovered from soil microcosms. We isolated several Ochrobactrum sp. and Pedobacter sp., and demonstrated that they inhabit the nematode gut by FISH. Although their role in A. maximus is not resolved, we propose possible mutualistic roles for these bacteria in protection of the host against pathogens and

  20. Revisiting a model-independent dark energy reconstruction method

    Energy Technology Data Exchange (ETDEWEB)

    Lazkoz, Ruth; Salzano, Vincenzo; Sendra, Irene [Euskal Herriko Unibertsitatea, Fisika Teorikoaren eta Zientziaren Historia Saila, Zientzia eta Teknologia Fakultatea, Bilbao (Spain)

    2012-09-15

    In this work we offer new insights into the model-independent dark energy reconstruction method developed by Daly and Djorgovski (Astrophys. J. 597:9, 2003; Astrophys. J. 612:652, 2004; Astrophys. J. 677:1, 2008). Our results, using updated SNeIa and GRBs, allow to highlight some of the intrinsic weaknesses of the method. Conclusions on the main dark energy features as drawn from this method are intimately related to the features of the samples themselves, particularly for GRBs, which are poor performers in this context and cannot be used for cosmological purposes, that is, the state of the art does not allow to regard them on the same quality basis as SNeIa. We find there is a considerable sensitivity to some parameters (window width, overlap, selection criteria) affecting the results. Then, we try to establish what the current redshift range is for which one can make solid predictions on dark energy evolution. Finally, we strengthen the former view that this model is modest in the sense it provides only a picture of the global trend and has to be managed very carefully. But, on the other hand, we believe it offers an interesting complement to other approaches, given that it works on minimal assumptions. (orig.)

  1. [Genetic regulation of T-lymphocyte responsiveness to PHA is independent of culture conditions (author's transl)].

    Science.gov (United States)

    Stiffel, C; Liacopoulos-Briot, M; Decreusefond, C; Lambert, F

    1979-01-01

    A maximal interline separation has been obtained after 10 consecutive generations of selective breeding for the character "quantitative in vitro response of lymph node lymphocytes to the mitogenic effect of phytohaemagglutinin". At the selection limit the difference between high and low responder lines was about 20-fold. A similar interline separation has been demonstrated for the T-mitogen effect of concanavalin A. The identical response to PPD (purified protein derivative of tuberculin), a B mitogen, proved that the genetic selection has only modified the potentialities of T lymphocytes. During the selective breeding, responsiveness to PHA stimulation has been always measured under identical culture conditions. To demonstrate that the interline difference in responsiveness was due essentially to genetic factors independent of environmental effects, a systematic study of various culture conditions has been undertaken. The optimal stimulation was found after two days of culture for high line cells and after three days for low line cells. The difference between maximal responses was only slightly lower than that obtained after a two-day culture as used for the selection test. Increase in cell concentrations produced higher thymidine incorporation. In the two lines, a linear correlation was established between the cell concentration and the response produced. The maximal response given by the highest number of low line lymphocytes was equivalent to that given by a number, 11-fold smaller, of high line cells. Within certain limits, changes in the amount of tritiated thymidine added to the culture did not affect the interline separation. With a thymidine of high specific activity, a sub-evaluation of uptake by high line cells decreased the interline difference. Results in mixed culture of lymph node cells from high and low lines indicated that the low response was not due to the release of inhibiting factors or to the presence of suppressive cells in low responder mice

  2. Protocol independent transmission method in software defined optical network

    Science.gov (United States)

    Liu, Yuze; Li, Hui; Hou, Yanfang; Qiu, Yajun; Ji, Yuefeng

    2016-10-01

    With the development of big data and cloud computing technology, the traditional software-defined network is facing new challenges (e.i., ubiquitous accessibility, higher bandwidth, more flexible management and greater security). Using a proprietary protocol or encoding format is a way to improve information security. However, the flow, which carried by proprietary protocol or code, cannot go through the traditional IP network. In addition, ultra- high-definition video transmission service once again become a hot spot. Traditionally, in the IP network, the Serial Digital Interface (SDI) signal must be compressed. This approach offers additional advantages but also bring some disadvantages such as signal degradation and high latency. To some extent, HD-SDI can also be regard as a proprietary protocol, which need transparent transmission such as optical channel. However, traditional optical networks cannot support flexible traffics . In response to aforementioned challenges for future network, one immediate solution would be to use NFV technology to abstract the network infrastructure and provide an all-optical switching topology graph for the SDN control plane. This paper proposes a new service-based software defined optical network architecture, including an infrastructure layer, a virtualization layer, a service abstract layer and an application layer. We then dwell on the corresponding service providing method in order to implement the protocol-independent transport. Finally, we experimentally evaluate that proposed service providing method can be applied to transmit the HD-SDI signal in the software-defined optical network.

  3. Culture-independent analysis of bacterial diversity in a child-care facility

    OpenAIRE

    Tin Sara; Lee Lesley; Kelley Scott T

    2007-01-01

    Abstract Background Child-care facilities appear to provide daily opportunities for exposure and transmission of bacteria and viruses. However, almost nothing is known about the diversity of microbial contamination in daycare facilities or its public health implications. Recent culture-independent molecular studies of bacterial diversity in indoor environments have revealed an astonishing diversity of microorganisms, including opportunistic pathogens and many uncultured bacteria. In this stud...

  4. On the number of independent cultural traits carried by individuals and populations

    Science.gov (United States)

    Lehmann, Laurent; Aoki, Kenichi; Feldman, Marcus W.

    2011-01-01

    In species subject to individual and social learning, each individual is likely to express a certain number of different cultural traits acquired during its lifetime. If the process of trait innovation and transmission reaches a steady state in the population, the number of different cultural traits carried by an individual converges to some stationary distribution. We call this the trait-number distribution. In this paper, we derive the trait-number distributions for both individuals and populations when cultural traits are independent of each other. Our results suggest that as the number of cultural traits becomes large, the trait-number distributions approach Poisson distributions so that their means characterize cultural diversity in the population. We then analyse how the mean trait number varies at both the individual and population levels as a function of various demographic features, such as population size and subdivision, and social learning rules, such as conformism and anti-conformism. Diversity at the individual and population levels, as well as at the level of cultural homogeneity within groups, depends critically on the details of population demography and the individual and social learning rules. PMID:21199846

  5. On the number of independent cultural traits carried by individuals and populations.

    Science.gov (United States)

    Lehmann, Laurent; Aoki, Kenichi; Feldman, Marcus W

    2011-02-12

    In species subject to individual and social learning, each individual is likely to express a certain number of different cultural traits acquired during its lifetime. If the process of trait innovation and transmission reaches a steady state in the population, the number of different cultural traits carried by an individual converges to some stationary distribution. We call this the trait-number distribution. In this paper, we derive the trait-number distributions for both individuals and populations when cultural traits are independent of each other. Our results suggest that as the number of cultural traits becomes large, the trait-number distributions approach Poisson distributions so that their means characterize cultural diversity in the population. We then analyse how the mean trait number varies at both the individual and population levels as a function of various demographic features, such as population size and subdivision, and social learning rules, such as conformism and anti-conformism. Diversity at the individual and population levels, as well as at the level of cultural homogeneity within groups, depends critically on the details of population demography and the individual and social learning rules.

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

    Directory of Open Access Journals (Sweden)

    R. Urbano

    2010-08-01

    Full Text Available 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.

  7. Determining $H_0$ with a model-independent method

    CERN Document Server

    Wu, Puxun; Yu, Hongwei

    2015-01-01

    In this letter, by using the type Ia supernovae (SNIa) to provide the luminosity distance (LD) directly, which is dependent on the value of the Hubble constant $H_0= 100 h\\; {\\rm km\\; s^{-1}\\; Mpc^{-1}}$, and the angular diameter distance from galaxy clusters or baryon acoustic oscillations (BAOs) to give the derived LD according to the distance duality relation, we propose a model-independent method to determine $h$ from the fact that different observations should give the same LD at a redshift. Combining the Union 2.1 SNIa and galaxy cluster data, we obtain that at the $1\\sigma$ confidence level (CL) $h=0.589\\pm0.030$ for the sample of the elliptical $\\beta$ model for galaxy clusters, and $h=0.635\\pm0.029$ for that of the spherical $\\beta$ model. The former is smaller than the values from other observations, while the latter is consistent with the Planck result at the $1\\sigma$ CL and agrees very well with the value reconstructed directly from the $H(z)$ data. With the Union 2.1 SNIa and BAO measurements, a...

  8. Culture-Independent Evaluation of the Appendix and Rectum Microbiomes in Children with and without Appendicitis

    Science.gov (United States)

    Davenport, Katherine P.; Fraser, Claire M.; Sandler, Anthony D.; Zeichner, Steven L.

    2014-01-01

    Purpose The function of the appendix is largely unknown, but its microbiota likely contributes to function. Alterations in microbiota may contribute to appendicitis, but conventional culture studies have not yielded conclusive information. We conducted a pilot, culture-independent 16S rRNA-based microbiota study of paired appendix and rectal samples. Methods We collected appendix and rectal swabs from 21 children undergoing appendectomy, six with normal appendices and fifteen with appendicitis (nine perforated). After DNA extraction, we amplified and sequenced 16S rRNA genes and analyzed sequences using CLoVR. We identified organisms differing in relative abundance using ANOVA (pappendicitis vs. normal), and disease severity (perforated vs. non-perforated). Results We identified 290 taxa in the study's samples. Three taxa were significantly increased in normal appendices vs. normal rectal samples: Fusibacter (p = 0.009), Selenomonas (p = 0.026), and Peptostreptococcus (p = 0.049). Five taxa were increased in abundance in normal vs. diseased appendices: Paenibacillaceae (p = 0.005), Acidobacteriaceae GP4 (p = 0.019), Pseudonocardinae (p = 0.019), Bergeyella (p = 0.019) and Rhizobium (p = 0.045). Twelve taxa were increased in the appendices of appendicitis patients vs. normal appendix: Peptostreptococcus (p = 0.0003), Bilophila (p = 0.0004), Bulleidia (p = 0.012), Fusobacterium (p = 0.018), Parvimonas (p = 0.003), Mogibacterium (p = 0.012), Aminobacterium (p = 0.019), Proteus (p = 0.028), Actinomycineae (p = 0.028), Anaerovorax (p = 0.041), Anaerofilum (p = 0.045), Porphyromonas (p = 0.010). Five taxa were increased in appendices in patients with perforated vs. nonperforated appendicitis: Bulleidia (p = 0.004), Fusibacter (p = 0.005), Prevotella (p = 0.021), Porphyromonas (p = 0.030), Dialister (p = 0.035). Three taxa were increased in rectum samples of patients with

  9. Determining H 0 using a model-independent method

    Science.gov (United States)

    Wu, Pu-Xun; Li, Zheng-Xiang; Yu, Hong-Wei

    2017-02-01

    By using type Ia supernovae (SNIa) to provide the luminosity distance (LD) directly, which depends on the value of the Hubble constant H 0 = 100 h km • s-1 • Mpc-1, and the angular diameter distance from galaxy clusters or baryon acoustic oscillations (BAOs) to give the derived LD according to the distance duality relation, we propose a model-independent method to determine h from the fact that different observations should give the same LD at a given redshift. Combining the Sloan Digital Sky Survey II (SDSS-II) SNIa from the MLCS2k2 light curve fit and galaxy cluster data, we find that at the 1σ confidence level (CL), h = 0:5867 ± 0:0303 for the sample of the elliptical β model for galaxy clusters, and h = 0:6199 ± 0:0293 for that of the spherical β model. The former is smaller than the values from other observations, whereas the latter is consistent with the Planck result at the 2σ CL and agrees very well with the value reconstructed directly from the H( z) data. With the SDSS-II SNIa and BAO measurements, a tighter constraint, h = 0:6683 ± 0:0221, is obtained. For comparison, we also consider the Union 2.1 SNIa from the SALT2 light curve fitting. The results from the Union 2.1 SNIa are slightly larger than those from the SDSS-II SNIa, and the Union 2.1 SNIa + BAOs give the tightest value. We find that the values from SNIa + BAOs are quite consistent with those from the Planck and the BAOs, as well as the local measurement from Cepheids and very-low-redshift SNIa.

  10. Combining culture-dependent and -independent methodologies for estimation of richness of estuarine bacterioplankton consuming riverine dissolved organic matter.

    Science.gov (United States)

    Kisand, Veljo; Wikner, Johan

    2003-06-01

    Three different methods for analyzing natural microbial community diversity were combined to maximize an estimate of the richness of bacterioplankton catabolizing riverine dissolved organic matter (RDOM). We also evaluated the ability of culture-dependent quantitative DNA-DNA hybridization, a 16S rRNA gene clone library, and denaturing gradient gel electrophoresis (DGGE) to detect bacterial taxa in the same sample. Forty-two different cultivatable strains were isolated from rich and poor solid media. In addition, 50 unique clones were obtained by cloning of the bacterial 16S rDNA gene amplified by PCR from the community DNA into an Escherichia coli vector. Twenty-three unique bands were sequenced from 12 DGGE profiles, excluding a composite fuzzy band of the Cytophaga-Flavobacterium group. The different methods gave similar distributions of taxa at the genus level and higher. However, the match at the species level among the methods was poor, and only one species was identified by all three methods. Consequently, all three methods identified unique subsets of bacterial species, amounting to a total richness of 97 operational taxonomic units in the experimental system. The confidence in the results was, however, dependent on the current precision of the phylogenetic determination and definition of the species. Bacterial consumers of RDOM in the studied estuary were primarily both cultivatable and uncultivable taxa of the Cytophaga-Flavobacterium group, a concordant result among the methods applied. Culture-independent methods also suggested several not-yet-cultivated beta-proteobacteria to be RDOM consumers.

  11. Senescence Process in Primary Wilms' Tumor Cell Culture Induced by p53 Independent p21 Expression.

    Science.gov (United States)

    Theerakitthanakul, Korkiat; Khrueathong, Jeerasak; Kruatong, Jirasak; Graidist, Potchanapond; Raungrut, Pritsana; Kayasut, Kanita; Sangkhathat, Surasak

    2016-01-01

    Wilms tumor (WT) is an embryonal tumor occurring in developing kidney tissue. WT cells showing invasive cancer characteristics, also retain renal stem cell behaviours. In-vitro culture of WT is hampered by limited replicative potential. This study aimed to establish a longterm culture of WT cells to enable the study of molecular events to attempt to explain its cellular senescence. Primary cell cultures from fresh WT tumor specimen were established. Of 5 cultures tried, only 1 could be propagated for more than 7 passages. One culture, identified as PSU-SK-1, could be maintained > 35 passages and was then subjected to molecular characterization and evaluation for cancer characteristics. The cells consistently harbored concomitant mutations of CTNNB1 (Ser45Pro) and WT1 (Arg413Stop) thorough the cultivation. On Transwell invasion assays, the cells exhibited migration and invasion at 55% and 27% capability of the lung cancer cells, A549. On gelatin zymography, PSU-SK-1 showed high expression of the matrix metaloproteinase. The cells exhibited continuous proliferation with 24-hour doubling time until passages 28-30 when the growth slowed, showing increased cell size, retention of cells in G1/S proportion and positive β-galactosidase staining. As with those evidence of senescence in advanced cell passages, expression of p21 and cyclin D1 increased when the expression of β-catenin and its downstream protein, TCF, declined. There was also loss-of-expression of p53 in this cell line. In conclusion, cellular senescence was responsible for limited proliferation in the primary culture of WT, which was also associated with increased expression of p21 and was independent of p53 expression. Decreased activation of the Wnt signalling might explain the induction of p21 expression.

  12. The dopamine D4 receptor gene (DRD4) moderates cultural difference in independent versus interdependent social orientation.

    Science.gov (United States)

    Kitayama, Shinobu; King, Anthony; Yoon, Carolyn; Tompson, Steve; Huff, Sarah; Liberzon, Israel

    2014-06-01

    Prior research suggests that cultural groups vary on an overarching dimension of independent versus interdependent social orientation, with European Americans being more independent, or less interdependent, than Asians. Drawing on recent evidence suggesting that the dopamine D4 receptor gene (DRD4) plays a role in modulating cultural learning, we predicted that carriers of DRD4 polymorphisms linked to increased dopamine signaling (7- or 2-repeat alleles) would show higher levels of culturally dominant social orientations, compared with noncarriers. European Americans and Asian-born Asians (total N = 398) reported their social orientation on multiple scales. They were also genotyped for DRD4. As in earlier work, European Americans were more independent, and Asian-born Asians more interdependent. This cultural difference was significantly more pronounced for carriers of the 7- or 2-repeat alleles than for noncarriers. Indeed, no cultural difference was apparent among the noncarriers. Implications for potential coevolution of genes and culture are discussed.

  13. Study on the Independence Construction of Organizational Culture of Independent College%独立学院组织文化的独立性建设研究

    Institute of Scientific and Technical Information of China (English)

    郝一峰

    2012-01-01

      The paper according to the independent institute as the object of study, to organizational behavior in the tissue culture of this concept, in order to " independence " as the theme, the union current our country independent college in the organizational culture construction basic present situation and the basic problems encountered, according to the construction of organizational culture in general principle, principle of operation, as well as the independent institute independent the requirements of the development of tissue culture, independence’s construction necessity, feasibility and reality, and explore feasible construction path from the school, people, structure and other aspects, for the independent college in China holding mode features to provide management of cultural intelligence support.%  本文以独立学院为研究对象,以组织行为学中的组织文化这一概念切入,以“独立性”作为主题,结合当前我国独立学院在组织文化建设的基本现状和遇到的基本问题,根据组织文化建设的一般运行原理、原则,以及独立学院独立性发展的要求,探讨组织文化独立性建设的必要性、可行性和现实性,并从校、人、制等多个方面入手,探索出具有可操作性的建设路径,为我国独立学院保持办学模式特色化提供管理文化层面上的智力支持。

  14. Independent data validation of an in vitro method for ...

    Science.gov (United States)

    In vitro bioaccessibility assays (IVBA) estimate arsenic (As) relative bioavailability (RBA) in contaminated soils to improve the accuracy of site-specific human exposure assessments and risk calculations. For an IVBA assay to gain acceptance for use in risk assessment, it must be shown to reliably predict in vivo RBA that is determined in an established animal model. Previous studies correlating soil As IVBA with RBA have been limited by the use of few soil types as the source of As. Furthermore, the predictive value of As IVBA assays has not been validated using an independent set of As-contaminated soils. Therefore, the current study was undertaken to develop a robust linear model to predict As RBA in mice using an IVBA assay and to independently validate the predictive capability of this assay using a unique set of As-contaminated soils. Thirty-six As-contaminated soils varying in soil type, As contaminant source, and As concentration were included in this study, with 27 soils used for initial model development and nine soils used for independent model validation. The initial model reliably predicted As RBA values in the independent data set, with a mean As RBA prediction error of 5.3% (range 2.4 to 8.4%). Following validation, all 36 soils were used for final model development, resulting in a linear model with the equation: RBA = 0.59 * IVBA + 9.8 and R2 of 0.78. The in vivo-in vitro correlation and independent data validation presented here provide

  15. Rapid culture-independent microbial analysis aboard the International Space Station (ISS).

    Science.gov (United States)

    Maule, Jake; Wainwright, Norm; Steele, Andrew; Monaco, Lisa; Morris, Heather; Gunter, Daniel; Damon, Michael; Wells, Mark

    2009-10-01

    A new culture-independent system for microbial monitoring, called the Lab-On-a-Chip Application Development Portable Test System (LOCAD-PTS), was operated aboard the International Space Station (ISS). LOCAD-PTS was launched to the ISS aboard Space Shuttle STS-116 on December 9, 2006, and has since been used by ISS crews to monitor endotoxin on cabin surfaces. Quantitative analysis was performed within 15 minutes, and sample return to Earth was not required. Endotoxin (a marker of Gram-negative bacteria) was distributed throughout the ISS, despite previous indications that mostbacteria on ISS surfaces were Gram-positive [corrected].Endotoxin was detected at 24 out of 42 surface areas tested and at every surface site where colony-forming units (cfu) were observed, even at levels of 4-120 bacterial cfu per 100 cm(2), which is below NASA in-flight requirements (3.78 EU per 100 cm(2)). Based upon data collected from the ISS so far, new culture-independent requirements (defined in EU) are suggested, which are verifiable in flight with LOCAD-PTS yet high enough to avoid false alarms. The suggested requirements are intended to supplement current ISS requirements (defined in cfu) and would serve a dual purpose of safeguarding crew health (internal spacecraft surfaces <20 EU per 100 cm(2)) and monitoring forward contamination during Constellation missions (surfaces periodically exposed to the external environment, including the airlock and space suits, <0.24 EU per 100 cm(2)).

  16. Male facial attractiveness and masculinity may provide sex- and culture-independent cues to semen quality.

    Science.gov (United States)

    Soler, C; Kekäläinen, J; Núñez, M; Sancho, M; Álvarez, J G; Núñez, J; Yaber, I; Gutiérrez, R

    2014-09-01

    Phenotype-linked fertility hypothesis (PLFH) predicts that male secondary sexual traits reveal honest information about male fertilization ability. However, PLFH has rarely been studied in humans. The aim of the present study was to test PLFH in humans and to investigate whether potential ability to select fertile partners is independent of sex or cultural background. We found that on the contrary to the hypothesis, facial masculinity was negatively associated with semen quality. As increased levels of testosterone have been demonstrated to impair sperm production, this finding may indicate a trade-off between investments in secondary sexual signalling (i.e. facial masculinity) and fertility or status-dependent differences in investments in semen quality. In both sexes and nationalities (Spanish and Colombian), ranked male facial attractiveness predicted male semen quality. However, Spanish males and females estimated facial images generally more attractive (gave higher ranks) than Colombian raters, and in both nationalities, males gave higher ranks than females. This suggests that male facial cues may provide culture- and sex-independent information about male fertility. However, our results also indicate that humans may be more sensitive to facial attractiveness cues within their own populations and also that males may generally overestimate the attractiveness of other men to females. © 2014 The Authors. Journal of Evolutionary Biology © 2014 European Society For Evolutionary Biology.

  17. Watermark Detection and Extraction Using Independent Component Analysis Method

    Directory of Open Access Journals (Sweden)

    Yu Dan

    2002-01-01

    Full Text Available This paper proposes a new image watermarking technique, which adopts Independent Component Analysis (ICA for watermark detection and extraction process (i.e., dewatermarking. Watermark embedding is performed in the spatial domain of the original image. Watermark can be successfully detected during the Principle Component Analysis (PCA whitening stage. A nonlinear robust batch ICA algorithm, which is able to efficiently extract various temporally correlated sources from their observed linear mixtures, is used for blind watermark extraction. The evaluations illustrate the validity and good performance of the proposed watermark detection and extraction scheme based on ICA. The accuracy of watermark extraction depends on the statistical independence between the original, key and watermark images and the temporal correlation of these sources. Experimental results demonstrate that the proposed system is robust to several important image processing attacks, including some geometrical transformations—scaling, cropping and rotation, quantization, additive noise, low pass filtering, multiple marks, and collusion.

  18. Low Fluid Shear Culture of Staphylococcus Aureus Represses hfq Expression and Induces an Attachment-Independent Biofilm Phenotype

    Science.gov (United States)

    Ott, C. Mark; Castro, S. L.; Nickerson, C. A.; Nelman-Gonzalez, M.

    2011-01-01

    Background: The opportunistic pathogen, Staphylococcus aureus, experiences fluctuations in fluid shear during infection and colonization of a human host. Colonization frequently occurs at mucus membrane sites such as in the gastrointestinal tract where the bacterium may experience low levels of fluid shear. The response of S. aureus to low fluid shear remains unclear. Methods: S. aureus was cultured to stationary phase using Rotating-Wall Vessel (RWV) bioreactors which produce a physiologically relevant low fluid shear environment. The bacterial aggregates that developed in the RWV were evaluated by electron microscopy as well as for antibiotic resistance and other virulence-associated stressors. Genetic expression profiles for the low-shear cultured S. aureus were determined by microarray analysis and quantitative real-time PCR. Results: Planktonic S. aureus cultures in the low-shear environment formed aggregates completely encased in high amounts of extracellular polymeric substances. In addition, these aggregates demonstrated increased antibiotic resistance indicating attachment-independent biofilm formation. Carotenoid production in the low-shear cultured S. aureus was significantly decreased, and these cultures displayed an increased susceptibility to oxidative stress and killing by whole blood. The hfq gene, associated with low-shear growth in Gram negative organisms, was also found to be down-regulated in S. aureus. Conclusions: Collectively, this data suggests that S. aureus decreases virulence characteristics in favor of a biofilm-dwelling colonization phenotype in response to a low fluid shear environment. Furthermore, the identification of an Hfq response to low-shear culture in S. aureus, in addition to the previously reported responses in Gram negative organisms, strongly suggests an evolutionarily conserved response to mechanical stimuli among structurally diverse prokaryotes.

  19. Succession of Actinomycetes During Composting Proccess of Dairy-Farm Waste Investigated by Culture-Dependent and Independent Approaches

    Directory of Open Access Journals (Sweden)

    Mukhlissul Faatih1

    2015-11-01

    Full Text Available Mesophilic, thermophilic, and maturation phases were recognized in composting proccess. Temperaturechanges influence the microbial communities in compost within composting proccess. Actinomycetes account for alarger part of compost microbial population. The aim of this research was to study succession of actinomycetescommunity during composting of dairy-farm waste investigated by culture-dependent and independentapproaches.In culture-independent method, the succession of actinomycetes community was analyzed by nestedpolymerasechain reaction of ribosomal intergenic spacer (nested-PCR RISA using spesific primer F243 and primerR23S followed by a second PCR using primers F968 and R23S. In culture-dependent method actinomycetes fromcompost were isolated on selective media, starch-nitrate medium and humic-acid + vitamins medium. DNA ofactinomycetes was extracted and amplified by repetitive sequence-based PCR (rep-PCR using primer BOXA1R. Thebanding patterns were used to generate dendrograms by UPGMA clustering with NTSYS program. Microcosmcontaining sterile rice-straw and water which is inoculated with each actinomycetes isolates was used for examiningthe ability of each isolate in rice-straw degradation.The experiment results showed that succession of both bacteria and actinomycetes was occured withincomposting proccess of dairy-farm waste. Analysed by culture-independent method revealed that the highestcommunity of compost’s bacteria was on mesophilic, thermophilic, and maturation phases, respectively. WhereasPCR-nested RISA resulted the highest population of actinomycetes was on thermophilic, maturation, and mesophilicphases, respectively. By culture-dependent method was obtained 29 actinomycetes isolates from mesophilic phase,23 isolates from thermophilic phase, and 19 isolates from maturation phase. Genetic diversity analysis of the obtainedisolates showed the presence of phylogenetic grouping on each phase of composting proccess. This result

  20. Caveolae-dependent and -independent uptake of albumin in cultured rodent pulmonary endothelial cells.

    Directory of Open Access Journals (Sweden)

    Hui-Hua Li

    Full Text Available Although a critical role for caveolae-mediated albumin transcytosis in pulmonary endothelium is well established, considerably less is known about caveolae-independent pathways. In this current study, we confirmed that cultured rat pulmonary microvascular (RPMEC and pulmonary artery (RPAEC endothelium endocytosed Alexa488-labeled albumin in a saturable, temperature-sensitive mode and internalization resulted in co-localization by fluorescence microscopy with cholera B toxin and caveolin-1. Although siRNA to caveolin-1 (cav-1 in RPAEC significantly inhibited albumin uptake, a remnant portion of albumin uptake was cav-1-independent, suggesting alternative pathways for albumin uptake. Thus, we isolated and cultured mouse lung endothelial cells (MLEC from wild type and cav-1(-/- mice and noted that ~ 65% of albumin uptake, as determined by confocal imaging or live cell total internal reflectance fluorescence microscopy (TIRF, persisted in total absence of cav-1. Uptake of colloidal gold labeled albumin was evaluated by electron microscopy and demonstrated that albumin uptake in MLEC from cav-1(-/- mice was through caveolae-independent pathway(s including clathrin-coated pits that resulted in endosomal accumulation of albumin. Finally, we noted that albumin uptake in RPMEC was in part sensitive to pharmacological agents (amiloride [sodium transport inhibitor], Gö6976 [protein kinase C inhibitor], and cytochalasin D [inhibitor of actin polymerization] consistent with a macropinocytosis-like process. The amiloride sensitivity accounting for macropinocytosis also exists in albumin uptake by both wild type and cav-1(-/- MLEC. We conclude from these studies that in addition to the well described caveolar-dependent pulmonary endothelial cell endocytosis of albumin, a portion of overall uptake in pulmonary endothelial cells is cav-1 insensitive and appears to involve clathrin-mediated endocytosis and macropinocytosis-like process.

  1. Culture-independent evaluation of the appendix and rectum microbiomes in children with and without appendicitis.

    Directory of Open Access Journals (Sweden)

    Hope T Jackson

    Full Text Available PURPOSE: The function of the appendix is largely unknown, but its microbiota likely contributes to function. Alterations in microbiota may contribute to appendicitis, but conventional culture studies have not yielded conclusive information. We conducted a pilot, culture-independent 16S rRNA-based microbiota study of paired appendix and rectal samples. METHODS: We collected appendix and rectal swabs from 21 children undergoing appendectomy, six with normal appendices and fifteen with appendicitis (nine perforated. After DNA extraction, we amplified and sequenced 16S rRNA genes and analyzed sequences using CLoVR. We identified organisms differing in relative abundance using ANOVA (p<0.05 by location (appendix vs. rectum, disease (appendicitis vs. normal, and disease severity (perforated vs. non-perforated. RESULTS: We identified 290 taxa in the study's samples. Three taxa were significantly increased in normal appendices vs. normal rectal samples: Fusibacter (p = 0.009, Selenomonas (p = 0.026, and Peptostreptococcus (p = 0.049. Five taxa were increased in abundance in normal vs. diseased appendices: Paenibacillaceae (p = 0.005, Acidobacteriaceae GP4 (p = 0.019, Pseudonocardinae (p = 0.019, Bergeyella (p = 0.019 and Rhizobium (p = 0.045. Twelve taxa were increased in the appendices of appendicitis patients vs. normal appendix: Peptostreptococcus (p = 0.0003, Bilophila (p = 0.0004, Bulleidia (p = 0.012, Fusobacterium (p = 0.018, Parvimonas (p = 0.003, Mogibacterium (p = 0.012, Aminobacterium (p = 0.019, Proteus (p = 0.028, Actinomycineae (p = 0.028, Anaerovorax (p = 0.041, Anaerofilum (p = 0.045, Porphyromonas (p = 0.010. Five taxa were increased in appendices in patients with perforated vs. nonperforated appendicitis: Bulleidia (p = 0.004, Fusibacter (p = 0.005, Prevotella (p = 0.021, Porphyromonas (p = 0.030, Dialister (p = 0.035. Three taxa were

  2. A generation method for a kind of linear independentLatin arrays

    Institute of Scientific and Technical Information of China (English)

    陶仁骥; 陈世华

    1995-01-01

    The concept of correlation of Latin array is introduced and the generation problem of Latin arrays with independence degree ≥1 is discussed. A method is given to generate such Latin arrays, that is, shifting invertible Boolean vector functions with independence degree ≥1. Finally, a generation method of linearly independent invertible Boolean vector functions is given.

  3. Method and Practice of Cultural Translation

    Institute of Scientific and Technical Information of China (English)

    Zhang; Hui

    2015-01-01

    In recent years,translation theorists begin to take translation as an activity of cultural exchange,that is,it is not only a linguistic transformation,but essentially a cultural communication.Language and culture are interactive.Language,as the carrier of the culture,can break through the limitation of time and space and carry all the aspects of the culture information.As a main means of cross-cultural communication,translation is connected with the culture tightly.However,in cross-cultural translation,especially between those cultures with great difference like Chinese and English,many cultural elements surface from the confrontation of cultural deposits,making it impossible to capture the relationship of the source text to the source culture in the target text.Thus,problems in translating cultural elements arise.Different approaches shall be adopted to solve this culture barrier.They are omission,transliteration,alienation and domestication.Using what kind of solutions is determined by many complicated factors.Under the circumstance of cultural globalization,these solutions are supposed to complement each other.Thus the translation action can become more scientific.

  4. Method and Practice of Cultural Translation

    Institute of Scientific and Technical Information of China (English)

    Zhang Hui

    2015-01-01

    In recent years,translation theorists begin to take translation as an activity of cultural exchange,that is,it is not only a linguistic transformation,but essentially a cultural communication.Language and culture are interactive.Language,as the carrier of the culture,can break through the limitation of time and space and carry all the aspects of the culture information. As a main meansof cross-cultural communication,translation is connected with the culture tightly.However,in cross-cultural translation,especially between those cultures with great difference like Chinese and English,many cultural elements surface from the confrontation of cultural deposits,making it impossible to capture the relationship of the source text to the source culture in the target text.Thus,problems in translating cultural elements arise. Different approaches shall be adopted to solve this culture barrier. They are omission,transliteration,alienation and domestication. Using what kind of solutions is determined by many complicated factors.Under the circumstance of cultural globalization,these solutions are supposed to complement each other.Thus the translation action can become more scientific.

  5. A Clash of Cultures: Improving the "Fit" between Evaluative Independence and the Political Requirements of a Democratic Society

    Science.gov (United States)

    Chelimsky, Eleanor

    2008-01-01

    This article presents a plenary address wherein the author talks about cultural clashes, about what happens when evaluation meets politics. In her address, the author talks about the kinds of clashes that occur on a regular basis between evaluative independence and the political culture it challenges, along with possible ways to predict, parry, or…

  6. A Clash of Cultures: Improving the "Fit" between Evaluative Independence and the Political Requirements of a Democratic Society

    Science.gov (United States)

    Chelimsky, Eleanor

    2008-01-01

    This article presents a plenary address wherein the author talks about cultural clashes, about what happens when evaluation meets politics. In her address, the author talks about the kinds of clashes that occur on a regular basis between evaluative independence and the political culture it challenges, along with possible ways to predict, parry, or…

  7. Revisiting bovine pyometra-New insights into the disease using a culture-independent deep sequencing approach

    DEFF Research Database (Denmark)

    Knudsen, Lif Rødtness Vesterby; Karstrup, Cecilia Christensen; Pedersen, Hanne Gervi;

    2015-01-01

    The bacteria present in the uterus during pyometra have previously been studied using bacteriological culturing. These studies identified Fusobacterium necrophorum and Trueperella pyogenes as the major contributors to the pathogenesis of pyometra. However, an increasing number of culture-independ...... rights reserved....

  8. Two-level method with coarse space size independent convergence

    Energy Technology Data Exchange (ETDEWEB)

    Vanek, P.; Brezina, M. [Univ. of Colorado, Denver, CO (United States); Tezaur, R.; Krizkova, J. [UWB, Plzen (Czech Republic)

    1996-12-31

    The basic disadvantage of the standard two-level method is the strong dependence of its convergence rate on the size of the coarse-level problem. In order to obtain the optimal convergence result, one is limited to using a coarse space which is only a few times smaller than the size of the fine-level one. Consequently, the asymptotic cost of the resulting method is the same as in the case of using a coarse-level solver for the original problem. Today`s two-level domain decomposition methods typically offer an improvement by yielding a rate of convergence which depends on the ratio of fine and coarse level only polylogarithmically. However, these methods require the use of local subdomain solvers for which straightforward application of iterative methods is problematic, while the usual application of direct solvers is expensive. We suggest a method diminishing significantly these difficulties.

  9. Rapid Culture-Independent Microbial Analysis Aboard the International Space Station (ISS)

    Science.gov (United States)

    Maule, Jake; Wainwright, Norm; Steele, Andrew; Monaco, Lisa; Morris, Heather; Gunter, Daniel; Damon, Michael; Wells, Mark

    2009-10-01

    A new culture-independent system for microbial monitoring, called the Lab-On-a-Chip Application Development Portable Test System (LOCAD-PTS), was operated aboard the International Space Station (ISS). LOCAD-PTS was launched to the ISS aboard Space Shuttle STS-116 on December 9, 2006, and has since been used by ISS crews to monitor endotoxin on cabin surfaces. Quantitative analysis was performed within 15 minutes, and sample return to Earth was not required. Endotoxin (a marker of Gram-negative bacteria and fungi) was distributed throughout the ISS, despite previous indications that most bacteria on ISS surfaces were Gram-positive. Endotoxin was detected at 24 out of 42 surface areas tested and at every surface site where colony-forming units (cfu) were observed, even at levels of 4-120 bacterial cfu per 100 cm2, which is below NASA in-flight requirements (dining facilities. Endotoxin was absent from airlock surfaces, except the Extravehicular Hatch Handle (>3.78 EU per 100 cm2). Based upon data collected from the ISS so far, new culture-independent requirements (defined in EU) are suggested, which are verifiable in flight with LOCAD-PTS yet high enough to avoid false alarms. The suggested requirements are intended to supplement current ISS requirements (defined in cfu) and would serve a dual purpose of safeguarding crew health (internal spacecraft surfaces <20 EU per 100 cm2) and monitoring forward contamination during Constellation missions (surfaces periodically exposed to the external environment, including the airlock and space suits, <0.24 EU per 100 cm2).

  10. Culture-dependent and -independent molecular analysis of the bacterial community within uranium ore.

    Science.gov (United States)

    Islam, Ekramul; Sar, Pinaki

    2011-08-01

    The bacterial community structure within a uranium ore was investigated using culture-dependent and -independent clone library analysis and denaturing gradient gel electrophoresis of 16S rRNA genes. The major aerobic heterotrophic bacteria were isolated and identified, and their resistance to uranium and other heavy metals was characterized. Together with near neutral pH, moderate organic carbon content, elevated U and other heavy metals (V, Ni, Mn, Cu, etc.), the ore showed high microbial counts and phylotype richness. The bacterial community mainly consisted of uncultured Proteobacteria, with the predominance of γ - over β - and α -subdivisions, along with Actinobacteria and Firmicutes. A phylogenetic study revealed that nearly one-third of the community was affiliated to as yet uncultured and unidentified bacteria having a closer relationship to Pseudomonas. Lineages of Burkholderiaceae and Moraxellaceae were relatively more abundant in the total community, while genera affiliated to Xanthomonadaceae and Microbacteriaceae and Exiguobacterium were detected in the culturable fraction. More than 50% of the bacterial isolates affiliated to Stenotrophomonas, Microbacterium, Acinetobacter, Pseudomonas and Enterobacter showed resistance to uranium and other heavy metals. The study showed for the first time that uranium ore harbors major bacterial groups related to organisms having a wide range of environmentally significant functional attributes, and the most abundant members are possibly new groups/taxa. These findings provide new insights into U-ore geomicrobiology that could be useful in biohydrometallurgy and bioremediation applications.

  11. Analysis of bacterial community in bulking sludge using culture-dependent and -independent approaches

    Institute of Scientific and Technical Information of China (English)

    Decai Jin; Ping Wang; Zhihui Bai; Xinxin Wang; Hong Peng; Rong Qi; Zhisheng Yu; Guoqiang Zhuang

    2011-01-01

    The bacterial community of a bulking sludge from a municipal wastewater treatment plant with anoxic-anaerobic-oxic process was investigated by combination of cultivation and 16S rRNA gene clone library analysis for understanding the causes of bulking.A total of 28 species were obtained from 63 isolates collected from six culture media.The most cultivable species belonged to γ-Proteobacteria including Klebsiella sp.,Pseudomonas sp.,Aeromonas sp.and Acinetobacter sp.Further analysis of these strains by repetitive sequence based on polymerase chain reaction (rep-PCR) technology showed that rep-PCR yielded discriminatory banding patterns within the same genus using REP and BOX primer sets.While the culture-independent assessment revealed that β-Proteobacteria was the dominant group in the bulking sample.Sequence analysis revealed that the highest proportion (14.7%) of operational taxonomic units was 98% similar to Candidatus Accumulibacter phosphatis,which is used to remove phosphorous from wastewater.Our results indicated that combining different approaches can produce complementary information,thus generate a more accurate view of microbial community in bulking sludge.

  12. Arteries respond to independent control of circumferential and shear stress in organ culture.

    Science.gov (United States)

    Wayman, Brian H; Taylor, W Robert; Rachev, Alexander; Vito, Raymond P

    2008-05-01

    Arteries respond to changes in global mechanical parameters (pressure, flow rate, and longitudinal stretching) by remodeling to restore local parameters (circumferential stress, shear stress, and axial strain) to baseline levels. Because a change in a single global parameter results in changes of multiple local parameters, the effects of individual local parameters on remodeling remain unknown. This study uses a novel approach to study remodeling in organ culture based on independent control of local mechanical parameters. The approach is illustrated by studying the short term effects of circumferential and shear stress on remodeling-related biological markers. Porcine carotid arteries were cultured for 3 days at a circumferential stress of 50 or 150 kPa or, in separate experiments, a shear stress of 0.75 or 2.25 Pa. At high circumferential stress, matrix synthesis, smooth muscle cell proliferation, and cell death are significantly greater, but matrix metalloproteinase-2 (MMP-2) and pro-MMP-2 activity are significantly less. In contrast, biological markers measured were unaffected by shear stress. Applications of the proposed approach for improved understanding of remodeling, optimizing mechanical conditioning of tissue engineered arteries, and selection of experimentally motivated growth laws are discussed.

  13. Culture-dependent and -independent approaches establish the complexity of a PAH-degrading microbial consortium

    Energy Technology Data Exchange (ETDEWEB)

    Vinas, M.; Sabate, J.; Solanas, A.M. [Barcelona Univ., Barcelona (Spain). Dept. of Microbiology; Guasp, C.; Lalucat, J. [Illes Balears Univ., Palma de Mallorca (Spain). Dept. of Biology

    2005-11-15

    Microbial consortia are used in the decontamination of polluted environmental sites. A microbial consortium obtained by batch enrichment culture is a closed system with controlled conditions in which micro-organisms with a potentially high growth rate are selected and become dominant. The aim of this study was to identify the members of consortium AM, in which earlier batch enrichment work had shown high biodegradation rates of the aromatic fraction of polycyclic aromatic hydrocarbon (PAH). The AM consortium was obtained by sequential enrichment in liquid culture with a PAH mixture of 3- and 4- ringed PAHs as the sole source of carbon and energy. The consortium was examined using a triple approach method based on various cultivation strategies, denaturing gradient electrophoresis (DGGE) and the screening of 16S and 18S rRNA gene clone libraries. Eleven different sequences by culture-dependent techniques and 7 by both DGGE and clone libraries were obtained, yielding 19 different microbial components. Proteobacteria were the dominant group, representing 83 per cent of the total, while the Cytophaga-Flexibactor-Bacteroides group (CFB) was 11 per cent, and Ascomycota fungi were 6 per cent. It was determined that {beta}-Proteobacteria were predominant in the DGGE and clone library methods, whereas they were a minority in culturable strains. The highest diversity and number of noncoincident sequences was achieved by the cultivation method that showed members of the {alpha},{beta}, and {gamma}-Proteobacteria, CFB bacterial group, and Ascomycota fungi. Only 6 of the 11 strains isolated showed PAH-degrading capability. The bacterial strain (AMS7) and the fungal strain (AMF1) achieved the greatest PAH depletion. Results indicated that polyphasic assessment is necessary for a proper understanding of the composition of a microbial consortium. It was concluded that microbial consortia are more complex than previously realized. 54 refs., 3 tabs., 3 figs.

  14. Culture-independent qunatification of Salmonella enterica in carcass gauze swabs by flotation prior to real-time PCR

    DEFF Research Database (Denmark)

    Löfström, Charlotta; Schelin, Jenny; Norling, Börje;

    2010-01-01

    , for culture-independent quantification of intact Salmonella in pig carcass gauze swabs (100 cm2) prior to quantitative PCR (qPCR). A novel approach was investigated, excluding the homogenization step prior to flotation, to improve the detection limit and speed up the quantification procedure. The buoyant....... The method allowed accurate quantification from 4.4×102 to at least 2.2×107 CFU Salmonella per swab sample using qPCR (without preceding DNA extraction) or selective plating on xylose lysine deoxycholate agar. Samples with 50 CFU could be detected occasionally but fell outside the linear range...... of the standard curve. The swab samples showed a broad biological diversity; for seven samples not inoculated with Salmonella, the microbial background flora (BGF) was determined to 5.0 ± 2.2 log CFU/ml sample withdrawn after flotation. It was determined that the proceeding PCR step was inhibited by BGF...

  15. Culture-Independent Metagenomic Surveillance of Commercially Available Probiotics with High-Throughput Next-Generation Sequencing.

    Science.gov (United States)

    Patro, Jennifer N; Ramachandran, Padmini; Barnaba, Tammy; Mammel, Mark K; Lewis, Jada L; Elkins, Christopher A

    2016-01-01

    Millions of people consume dietary supplements either following a doctor's recommendation or at their own discretion to improve their overall health and well-being. This is a rapidly growing trend, with an associated and expanding manufacturing industry to meet the demand for new health-related products. In this study, we examined the contents and microbial viability of several popular probiotic products on the United States market. Culture-independent methods are proving ideal for fast and efficient analysis of foodborne pathogens and their associated microbial communities but may also be relevant for analyzing probiotics containing mixed microbial constituents. These products were subjected to next-generation whole-genome sequencing and analyzed by a custom in-house-developed k-mer counting method to validate manufacturer label information. In addition, the batch variability of respective products was examined to determine if any changes in their formulations and/or the manufacturing process occurred. Overall, the products we tested adhered to the ingredient claims and lot-to-lot differences were minimal. However, there were a few discrepancies in the naming of closely related Lactobacillus and Bifidobacterium species, whereas one product contained an apparent Enterococcus contaminant in two of its three lots. With the microbial contents of the products identified, we used traditional PCR and colony counting methods to comparatively assess our results and verify the viability of the microbes in these products with regard to the labeling claims. Of all the supplements examined, only one was found to be inaccurate in viability. Our use of next-generation sequencing as an analytical tool clearly demonstrated its utility for quickly analyzing commercially available products containing multiple microbes to ensure consumer safety. IMPORTANCE The rapidly growing supplement industry operates without a formal premarket approval process. Consumers rely on product labels to

  16. A coverage independent method to analyze large scale anisotropies

    CERN Document Server

    Billoir, Pierre

    2007-01-01

    The arrival time distribution of cosmic ray events is well suited to extract information regarding sky anisotropies. For an experiment with nearly constant exposure, the frequency resolution one can achieve is given by the inverse of the time $T$ during which the data was recorded. For $T$ larger than one calendar year the resolution becomes sufficient to resolve the sidereal and diurnal frequencies. Using a Fourier expansion on a modified time parameter, we show in this note that one can accurately extract sidereal modulations without knowledge of the experimental coverage. This procedure also gives the full frequency pattern of the event sample under studies which contains important information about possible systematics entering in the sidereal analysis. We also show how this method allows to correct for those systematics. Finally, we show that a two dimensional analysis, in the form of the spherical harmonic ($Y_l^m$) decomposition, can be performed under the same conditions for all $m\

  17. Cultural Impact on Participatory Deisgn Method

    DEFF Research Database (Denmark)

    Jensen, Mika Yasuoka

    2012-01-01

    This is a position paper for Cultural Impact on User Experience Design and Evaluation - One day workshop, NordiCHI, 2012.......This is a position paper for Cultural Impact on User Experience Design and Evaluation - One day workshop, NordiCHI, 2012....

  18. Microbial dynamics and biodiversity of table olives fermentations: the different culture-dependent and –independent approaches

    Directory of Open Access Journals (Sweden)

    Cristian eBotta

    2012-07-01

    Full Text Available The microbial ecology of the table olive fermentation process is a complex set of dynamics in which the roles of the lactic acid bacteria (LAB and yeast populations are closely related, and this synergism is of fundamental importance to obtain high quality products. Several studies on the ecology of table olives, both in spontaneous fermentations and in inoculated ones, have focused on the identification and characterization of yeasts, as they play a key role in the definition of the final organoleptic profiles through the production of volatile compounds. Moreover, these are able to promote the growth of LAB, which is responsible for the stabilization of the final product through the acidification activity and the inhibition of the growth of pathogenic bacteria. The current empirical production process of table olives could be improved through the development of mixed starter cultures. These can only be developed after a deep study of the population dynamics of yeasts and LAB by means of molecular methods. Until now, most studies have exploited culture-dependent approaches to define the natural microbiota of brine and olives. These approaches have identified two main species of LAB, namely Lactobacillus plantarum and L. pentosus, while, as far as yeasts are concerned, the most frequently isolated genera are Candida, Pichia and Saccharomyces. However, there are a few studies in literature in which a culture-independent approach has been employed. This review summarizes the state of the art of the microbial ecology of table olive fermentations and it focuses on the different approaches and molecular methods that have been applied.

  19. Identifying the bacterial community on the surface of Intralox belting in a meat boning room by culture-dependent and culture-independent 16S rDNA sequence analysis.

    Science.gov (United States)

    Brightwell, Gale; Boerema, Jackie; Mills, John; Mowat, Eilidh; Pulford, David

    2006-05-25

    We examined the bacterial community present on an Intralox conveyor belt system in an operating lamb boning room by sequencing the 16S ribosomal DNA (rDNA) of bacteria extracted in the presence or absence of cultivation. RFLP patterns for 16S rDNA clone library and cultures were generated using HaeIII and MspI restriction endonucleases. 16S rDNA amplicons produced 8 distinct RFLP pattern groups. RFLP groups I-IV were represented in the clone library and RFLP groups I and V-VIII were represented amongst the cultured isolates. Partial DNA sequences from each RFLP group revealed that all group I, II and VIII representatives were Pseudomonas spp., group III were Sphingomonas spp., group IV clones were most similar to an uncultured alpha proteobacterium, group V was similar to a Serratia spp., group VI with an Alcaligenes spp., and group VII with Microbacterium spp. Sphingomonads were numerically dominant in the culture-independent clone library and along with the group IV alpha proteobacterium were not represented amongst the cultured isolates. Serratia, Alcaligenes and Microbacterium spp. were only represented with cultured isolates. Pseudomonads were detected by both culture-dependent (84% of isolates) and culture-independent (12.5% of clones) methods and their presence at high frequency does pose the risk of product spoilage if transferred onto meat stored under aerobic conditions. The detection of sphingomonads in large numbers by the culture-independent method demands further analysis because sphingomonads may represent a new source of meat spoilage that has not been previously recognised in the meat processing environment. The 16S rDNA collections generated by both methods were important at representing the diversity of the bacterial population associated with an Intralox conveyor belt system.

  20. Rapid method for culturing embryonic neuron-glial cell cocultures

    DEFF Research Database (Denmark)

    Svenningsen, Åsa Fex; Shan, Wei-Song; Colman, David R;

    2003-01-01

    A streamlined, simple technique for primary cell culture from E17 rat tissue is presented. In an attempt to standardize culturing methods for all neuronal cell types in the embryo, we evaluated a commercial medium without serum and used similar times for trypsinization and tested different surfaces...... for plating. In 1 day, using one method and a single medium, it is possible to produce robust E17 cultures of dorsal root ganglia (DRG), cerebellum, and enteric plexi. Allowing the endogenous glial cells to repopulate the cultures saves time compared with existing techniques, in which glial cells are added...... to cultures first treated with antimitotic agents. It also ensures that all the cells present in vivo will be present in the culture. Myelination commences after approximately 2 weeks in culture for dissociated DRG and 3-4 weeks in cerebellar cultures. In enteric cultures, glial wrapping of the enteric...

  1. REVIEW: Previous Deception detection methods and New proposed method using independent component analysis of EEG signals.

    Directory of Open Access Journals (Sweden)

    Roshni D. Tale

    2014-04-01

    Full Text Available Deception detection has important legal and medical applications, but the reliability of methods for the differentiation between truthful and deceptive responses is still limited. Deception detection can be more accurately achieved by measuring the brain correlates of lying in an individual. For the evaluation of the method, several participants were gone through the designed concealed information test paradigm and their respective brain signals were recorded. The electroencephalogram (EEG signals were recorded and separated into many single trials. To enhance signal noise ratio (SNR of P3 components, the independent component analysis (ICA method was adopted to separate non-P3 (i.e. artifacts and P3 components from every single trial. Then the P3 waveforms with high SNR were reconstructed. And then group of features based on time, frequency, and amplitude were extracted from the reconstructed P3 waveforms. Finally, two different class of feature samples were used to train a support vector machine (SVM classifier because it has higher performance compared with several other classifiers. The method presented in this paper improves the efficiency of CIT and deception detection in comparison with previous reported methods.

  2. Assessment of the microbial diversity at the surface of Livarot cheese using culture-dependent and independent approaches

    OpenAIRE

    Mounier, Jérôme; Monnet, Christophe; Jacques, Noémie; Antoinette, A.; Irlinger, Françoise

    2009-01-01

    International audience; The microbial diversity of the surface of a commercial red-smear cheese, Livarot cheese, sold on the retail market was studied using culture-dependent and independent approaches. Forty yeasts and 40 bacteria from the cheese surface were collected, dereplicated using single-strand conformation polymorphism (SSCP) analysis and identified using rRNA gene sequencing for the culture-dependent approach. The cultureindependent approach involved cloning and sequencing of the 1...

  3. Simple and convenient method for culturing anaerobic bacteria.

    OpenAIRE

    Behbehani, M J; Jordan, H. V.; Santoro, D L

    1982-01-01

    A simple and convenient method for culturing anaerobic bacteria is described. Cultures can be grown in commercially available flasks normally used for preparation of sterile external solutions. A special disposable rubber flask closure maintains anaerobic conditions in the flask after autoclaving. Growth of a variety of anaerobic oral bacteria was comparable to that obtained after anaerobic incubation of broth cultures in Brewer Anaerobic Jars.

  4. Culturing Schwann Cells from Neonatal Rats by Improved Enzyme Digestion Combined with Explants-culture Method.

    Science.gov (United States)

    Liu, Di; Liang, Xiao-Chun; Zhang, Hong

    2016-08-01

    Objective To develop an improved method for culturing Schwann cells(SCs) by using both enzyme digestion and explants-culture approaches and compared with traditional explants-culture method and general hemi-explants-culture method. Methods Bilaterally sciatic nerves and brachial plexus nerves were dissected from 3 to 5-day-old neonatal SD rats and explants-culture method,general hemi-explants-culture method,and improved enzyme digestion combined with explants-culture method were adopted to culture SCs,respectively. SCs were digested and passaged after 7 days in culture and counted under the microscope. The purity of SCs was identified by S-100 immunofluorescence staining. Results The SCs of improved method group grew fastest and the total number of cells obtained was(1.85±0.13)×10(6);the SCs of the hemi-explants-culture method group grew slower than the improved method group and the total number of cells obtained was (1.10±0.10)×10(6);the SCs of the explants-culture method group grew slowest and the total number of cells obtained was (0.77±0.03)×10(6).The total number of cells obtained showed significant difference among the three groups(Pculture method group,and (74.50±4.23)% in the explants-culture method group(Pculture method can obtain sufficient amount of high-purity SCs in a short time and thus may be applied in further research on peripheral nerve regeneration.

  5. Microfluidics and multielectrode array-compatible organotypic slice culture method.

    Science.gov (United States)

    Berdichevsky, Yevgeny; Sabolek, Helen; Levine, John B; Staley, Kevin J; Yarmush, Martin L

    2009-03-30

    Organotypic brain slice cultures are used for a variety of molecular, electrophysiological, and imaging studies. However, the existing culture methods are difficult or expensive to apply in studies requiring long-term recordings with multielectrode arrays (MEAs). In this work, a novel method to maintain organotypic cultures of rodent hippocampus for several weeks on standard MEAs in an unmodified tissue culture incubator is described. Polydimethylsiloxane (Sylgard) mini-wells were used to stabilize organotypic cultures on glass and MEA surfaces. Hippocampus slices were successfully maintained within PDMS mini-wells for multiple weeks, with preserved pyramidal layer organization, connectivity, and activity. MEAs were used to record the development of spontaneous activity in an organotypic cultures for 4 weeks. This method is compatible with integration of microchannels into the culture substrate. Microchannels were incorporated into the mini-wells and applied to the guidance of axons originating within the slice, paving the way for studies of axonal sprouting using organotypic slices.

  6. Diversity of bacteria and archaea from a landfill in Chandigarh, India as revealed by culture-dependent and culture-independent molecular approaches.

    Science.gov (United States)

    Krishnamurthi, S; Chakrabarti, T

    2013-02-01

    The bacterial community structure of a municipal landfill in Chandigarh, India was analysed by culture-dependent as well as culture-independent molecular approaches, and archaeal structure by the latter method. Samples were collected in two phases from the surface and a depth of 0.91 m in June, 2004 and from 0.91 m, 1.52 m and 1.68 m in May, 2005. After serial dilutions, samples were plated onto tryptic soy agar (TSA), plate count agar (PCA), tryptic soy broth agar (TSBA) and TSBA100 (TSBA diluted 100 times and solidified with agarose), and incubated aerobically at 30°C. The number of bacteria (CFU) on different media ranged between 9.4×10⁵g⁻¹ (on PCA) and 1.9×10⁷g⁻¹ (on TSA) (wet weight). The numbers of bacteria enumerated from plates incubated anaerobically (anaerobic agar and reinforced clostridial agar) were 2.1×10⁷and 1.7×10⁶g⁻¹, respectively. Of the 468 isolated and purified bacteria (183 in the first phase and 285 in the second phase), 135 were characterised using phenotypic characteristics as well as 16S rRNA gene sequence analysis. It was found that members of the phylum Firmicutes were overwhelmingly predominant (86.6%) in the landfill, followed by Actinobacteria (9.6%) and Proteobacteria (3.7%). Among the Firmicutes, at least 17 species from the single genus Bacillus were the most abundant inhabitants of the landfill. Detailed polyphasic characterisation of many of these isolates led to the discovery of a novel genus Paenisporosarcina (and the species P. quisquiliarum), a novel species of Microbacterium, M. immunditiarum, and reclassification of Sporosarcina macmurdoensis, Pelagibacillus goriensis, Bacillus silvestris, Bacillus insolitus, Bacillus psychrotolerans and Bacillus psychrodurans. Culture-independent analysis of two 16S rRNA gene libraries also revealed that the phylum Firmicutes was the predominant group in this community. The diversity of Archaea was found to be limited mainly to members of two orders: Methanosarcinales

  7. A Method, Computer Program and System for Inferring Relations Between Cultural Specific Concepts in Two Cultures

    DEFF Research Database (Denmark)

    2013-01-01

    The present invention relates to a method, computer program and system for inferring relations between cultural specific concepts (CSC) in two cultures at least comprising the steps of - extracting and listing said cultural specific concepts (CSCs) and features of said CSCs from at least a first...

  8. Exploring the sources of bacterial spoilers in beefsteaks by culture-independent high-throughput sequencing.

    Directory of Open Access Journals (Sweden)

    Francesca De Filippis

    Full Text Available Microbial growth on meat to unacceptable levels contributes significantly to change meat structure, color and flavor and to cause meat spoilage. The types of microorganisms initially present in meat depend on several factors and multiple sources of contamination can be identified. The aims of this study were to evaluate the microbial diversity in beefsteaks before and after aerobic storage at 4°C and to investigate the sources of microbial contamination by examining the microbiota of carcasses wherefrom the steaks originated and of the processing environment where the beef was handled. Carcass, environmental (processing plant and meat samples were analyzed by culture-independent high-throughput sequencing of 16S rRNA gene amplicons. The microbiota of carcass swabs was very complex, including more than 600 operational taxonomic units (OTUs belonging to 15 different phyla. A significant association was found between beef microbiota and specific beef cuts (P<0.01 indicating that different cuts of the same carcass can influence the microbial contamination of beef. Despite the initially high complexity of the carcass microbiota, the steaks after aerobic storage at 4°C showed a dramatic decrease in microbial complexity. Pseudomonas sp. and Brochothrix thermosphacta were the main contaminants, and Acinetobacter, Psychrobacter and Enterobacteriaceae were also found. Comparing the relative abundance of OTUs in the different samples it was shown that abundant OTUs in beefsteaks after storage occurred in the corresponding carcass. However, the abundance of these same OTUs clearly increased in environmental samples taken in the processing plant suggesting that spoilage-associated microbial species originate from carcasses, they are carried to the processing environment where the meat is handled and there they become a resident microbiota. Such microbiota is then further spread on meat when it is handled and it represents the starting microbial association

  9. Exploring the sources of bacterial spoilers in beefsteaks by culture-independent high-throughput sequencing.

    Science.gov (United States)

    De Filippis, Francesca; La Storia, Antonietta; Villani, Francesco; Ercolini, Danilo

    2013-01-01

    Microbial growth on meat to unacceptable levels contributes significantly to change meat structure, color and flavor and to cause meat spoilage. The types of microorganisms initially present in meat depend on several factors and multiple sources of contamination can be identified. The aims of this study were to evaluate the microbial diversity in beefsteaks before and after aerobic storage at 4°C and to investigate the sources of microbial contamination by examining the microbiota of carcasses wherefrom the steaks originated and of the processing environment where the beef was handled. Carcass, environmental (processing plant) and meat samples were analyzed by culture-independent high-throughput sequencing of 16S rRNA gene amplicons. The microbiota of carcass swabs was very complex, including more than 600 operational taxonomic units (OTUs) belonging to 15 different phyla. A significant association was found between beef microbiota and specific beef cuts (PPseudomonas sp. and Brochothrix thermosphacta were the main contaminants, and Acinetobacter, Psychrobacter and Enterobacteriaceae were also found. Comparing the relative abundance of OTUs in the different samples it was shown that abundant OTUs in beefsteaks after storage occurred in the corresponding carcass. However, the abundance of these same OTUs clearly increased in environmental samples taken in the processing plant suggesting that spoilage-associated microbial species originate from carcasses, they are carried to the processing environment where the meat is handled and there they become a resident microbiota. Such microbiota is then further spread on meat when it is handled and it represents the starting microbial association wherefrom the most efficiently growing microbial species take over during storage and can cause spoilage.

  10. Qualitative study of three cell culture methods.

    Science.gov (United States)

    Wang, Aiguo; Xia, Tao; Ran, Peng; Chen, Xuemin; Nuessler, Andreas K

    2002-01-01

    Primary rat hepatocytes were cultured using different in vitro models and the enzyme leakage, albumin secretion, and cytochrome P450 1A (CYP 1A) activity were observed. The results showed that the level of LDH was decreased over time in culture. However, on day 5, LDH showed a significant increase in monolayer culture (MC) while after day 8 no LDH was detectable in sandwich culture (SC). The levels of AST and ALT did not change significantly over the investigated time. The CYP 1A activity was gradually decreased in a time-dependent manner in MC and SC. The decline of CYP 1A was faster in MC than in SC. This effect was partially reversed by using cytochrome P450 (CYP450) inducer such as Omeprazol and 3-methylcholanthrene (3-MC) and the CYP 1A induction was always higher in MC than in SC. In bioreactor basic CYP 1A activity was preserved over 2 weeks and the highest albumin production was observed in bioreactor followed by SC and MC. Taken together, it was indicated each investigated model had its advantages and disadvantages. It was also underlined that various in vitro models may address different questions.

  11. Mouse cell culture - Methods and protocols

    Directory of Open Access Journals (Sweden)

    CarloAlberto Redi

    2010-12-01

    Full Text Available The mouse is, out of any doubt, the experimental animal par excellence for many many colleagues within the scientific community, notably for those working in mammalian biology (in a broad sense, from basic genetic to modeling human diseases, starting at least from 1664 Robert Hooke experiments on air’s propertyn. Not surprising then that mouse cell cultures is a well established field of research itself and that there are several handbooks devoted to this discipline. Here, Andrew Ward and David Tosh provide a necessary update of the protocols currently needed. In fact, nearly half of the book is devoted to stem cells culture protocols, mainly embryonic, from a list of several organs (kidney, lung, oesophagus and intestine, pancreas and liver to mention some........

  12. Mouse cell culture: methods and protocols

    OpenAIRE

    Elvira M. Guerra Shinohara

    2010-01-01

    The mouse is, out of any doubt, the experimental animal par excellence for many many colleagues within the scientific community, notably for those working in mammalian biology (in a broad sense, from basic genetic to modeling human diseases), starting at least from 1664 Robert Hooke experiments on air’s propertyn. Not surprising then that mouse cell cultures is a well established field of research itself and that there are several handbooks devoted to this discipline. Here, Andrew Ward ...

  13. A new method of heart sound signal analysis based on independent function element

    Directory of Open Access Journals (Sweden)

    Cheng Xie-feng

    2014-09-01

    Full Text Available In this paper, a new method is presented for heart sound signal processing in statistical domain. The multiple components obtained from the conventional linear transformation are possibly irrelevant, but usually do not possess the characteristics of statistical independence. First, the definition and obtaining method of independent function element are discussed; the method of signal decomposition and reconstruction based on the independent function element, not only inherits the advantages of linear transformation, but also has the capability of signal representation in the statistical domain. After that, the application of independent function element in heart sound signal analysis is analyzed in detail. The validity and practicability of the method are demonstrated through two experiments.

  14. The awareness of employees in safety culture through the improved nuclear safety culture evaluation method

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Young Ga; Sung, Chan Ho; Jung, Yeon Sub [KHNP Central Research Institute, Daejeon (Korea, Republic of)

    2012-10-15

    After the Chernobyl nuclear accident in 1986, nuclear safety culture terminology was at first introduced emphasizing the importance of employees' attitude and organizational safety. The concept of safety culture was spread by INSAG 4 published in 1991. From that time, IAEA had provided the service of ASCOT for the safety culture assessment. However, many people still are thinking that safety culture is abstract and is not clear. It is why the systematic and reliable assessment methodology was not developed. Assessing safety culture is to identify what is the basic assumption for any organization to accept unconsciously. Therefore, it is very difficult to reach a meaningful conclusion by a superficial investigation alone. KHNP had been doing the safety culture assessment which was based on ASCOT methodology every 2 years. And this result had contributed to improving safety culture. But this result could not represent the level of organization's safety culture due to the limitation of method. So, KHNP has improved the safety culture method by benchmarking the over sea assessment techniques in 2011. The effectiveness of this improved methodology was validated through a pilot assessment. In this paper, the level of employees' safety culture awareness was analyzed by the improved method and reviewed what is necessary for the completeness and objectivity of the nuclear safety culture assessment methodology.

  15. Explaining the Effectiveness of the Contrast Culture Method for Managing Interpersonal Interactions across Cultures

    Science.gov (United States)

    Hiratsuka, Hiroyoshi; Suzuki, Hanako; Pusina, Alexis

    2016-01-01

    One of the current challenges in the field of intercultural education comes from the limited availability of training efficacy studies. The present study focused on explaining the effectiveness of the Contrast Culture Method (CCM) as an intercultural education method for managing interpersonal interactions across cultures between graduate…

  16. Assessing the performance of the independence method in modeling spatial extreme rainfall

    Science.gov (United States)

    Zheng, Feifei; Thibaud, Emeric; Leonard, Michael; Westra, Seth

    2015-09-01

    Spatial statistical methods are often employed to improve precision when estimating marginal distributions of extreme rainfall. Methods such as max-stable and copula models parameterize the spatial dependence and provide a continuous spatial representation. Alternatively, the independence method can be used to estimate marginal parameters without the need for parameterizing the spatial dependence, and this method has been under-utilized in hydrologic applications. This paper investigates the effectiveness of the independence method for marginal parameter estimation of spatially dependent extremes. Its performance is compared with three spatial dependence models (max-stable Brown-Resnick, max-stable Schlather, and Gaussian copula) by means of a simulation study. The independence method is statistically robust in estimating parameters and their associated confidence intervals for spatial extremes with various underlying dependence structures. The spatial dependence models perform comparably with the independence method when the spatial dependence structure is correctly specified; otherwise they exhibit considerably worse performance. We conclude that the independence method is more appealing for modeling the marginal distributions of spatial extremes (e.g., regional estimation of trends in rainfall extremes) due to its greater robustness and simplicity. The four statistical methods are illustrated using a spatial data set comprising 69 subdaily rainfall series from the Greater Sydney region, Australia.

  17. Methodical support of assessment of enterprise corporate culture

    Directory of Open Access Journals (Sweden)

    M.I. Ovcharenko

    2013-06-01

    Full Text Available The aim of the article. The article summarizes the existing theoretical approaches to the assessment of corporate culture of enterprise and defined the benefits and shortcomings of existing assessment methods.The results of the analysis. In particular, the present methods of such complex phenomena as organization's culture are conventionally divided into three groups: holistic scholar is deeply immersed in the culture of the organization and acts as communication observer; metaphorical the researcher uses language samples of documents, reports that there are stories and conversations that help to identify the fingerprints of culture, quantitative the researcher evaluates many points of view for evaluating the attributes of organization's culture.The authors concluded that it is important to develop methodology based on empirical evidence obtained as a result of a combination of both quantitative and holistic techniques for a comprehensive assessment of corporate culture. It will maximize the assessment of corporate culture. Thorough analysis of existing scientific research and theoretical developments of the mentioned problems the authors revealed the absence of adequate methodological approaches to quantify the level of corporate culture in the company.When calculating the integral indicator of corporate culture LCC Level of corporate culture, the authors performed a mathematical formalization assessment of corporate culture, the algorithm of calculation which includes seven stages.Hierarchical three-level structure of corporate culture was developed for assessing the corporate culture of the company. The limits of each criterion were determined. For identifying the relationship of each qualitative assessment of the value of the i-th parameter with the appropriate level of corporate culture (low, medium, high, high authors used the method of analysis T.Saaty. Formula for calculating the integral index, defined by the range and quality level of

  18. High-Throughput Sequencing and Metagenomics: Moving Forward in the Culture-Independent Analysis of Food Microbial Ecology

    OpenAIRE

    Ercolini, Danilo

    2013-01-01

    Following recent trends in environmental microbiology, food microbiology has benefited from the advances in molecular biology and adopted novel strategies to detect, identify, and monitor microbes in food. An in-depth study of the microbial diversity in food can now be achieved by using high-throughput sequencing (HTS) approaches after direct nucleic acid extraction from the sample to be studied. In this review, the workflow of applying culture-independent HTS to food matrices is described. T...

  19. The Factor of Culture in Modernization fnd Predicting the Commonwealth of Independent States Future

    Directory of Open Access Journals (Sweden)

    K P Kurylev

    2012-03-01

    Full Text Available The article considers the integration processes features within the Commonwealth of Independent States, defines the role and place of cooperation in the humanitarian sphere. The authors make an attempt to predict the future of the CIS.

  20. An Optimized Culture Method of Rat Dorsal Root Ganglion Neurons

    Institute of Scientific and Technical Information of China (English)

    LIUYin; CHENJing-Hong; GONGZe-Hui

    2004-01-01

    AIM: To establish a primary culture technique of acutely isolated dorsal root ganglion (DRG) neurons, and provide a simple & useful in vitro model for study of analgesia. Methods: Acutely isolated dorsal root ganglion (DRG) neurons were planted and cultured; the configuration and growth characters of DRG neurons were observed through inverted microscope.

  1. Tissue-Culture Method of Cloning Rubber Plants

    Science.gov (United States)

    Ball, E. A.

    1983-01-01

    Guayule plant, a high-yield rubber plant cloned by tissue-culture method to produce multiple new plants that mature quickly. By adjusting culture medium, excised shoot tip produces up to 50 identical guayule plants. Varying concentration of cytokinin, single excised tip produces either 1 or several (up to 50) new plants.

  2. Comparing Three Methods to Create Multilingual Phone Models for Vocabulary Independent Speech Recognition Tasks

    Science.gov (United States)

    2000-08-01

    Glass, et al.: Multilingual Spoken Language Under- ( multilingual clusters) and 5280 monolingual clusters. This standing in the MIT VOYAGER System...UNCLASSIFIED Defense Technical Information Center Compilation Part Notice ADP010392 TITLE: Comparing Three Methods to Create Multilingual Phone...METHODS TO CREATE MULTILINGUAL PHONE MODELS FOR VOCABULARY INDEPENDENT SPEECH RECOGNITION TASKS Joachim Kdhler German National Research Center for

  3. Recommendations for Culturally Sensitive Research Methods

    Science.gov (United States)

    Awad, Germine H.; Patall, Erika A.; Rackley, Kadie R.; Reilly, Erin D.

    2016-01-01

    As the US continues to diversify, methods for accurately assessing human behavior must evolve. This paper offers multicultural research considerations at several stages of the research process for psychological research and consultation. Implications regarding the comparative research framework are discussed and suggestions are offered on how to…

  4. Recommendations for Culturally Sensitive Research Methods

    Science.gov (United States)

    Awad, Germine H.; Patall, Erika A.; Rackley, Kadie R.; Reilly, Erin D.

    2016-01-01

    As the US continues to diversify, methods for accurately assessing human behavior must evolve. This paper offers multicultural research considerations at several stages of the research process for psychological research and consultation. Implications regarding the comparative research framework are discussed and suggestions are offered on how to…

  5. Comparison of liquid culture methods and effect of temporary

    African Journals Online (AJOL)

    star-pc

    2012-04-24

    Apr 24, 2012 ... Four different liquids, as well as solid culture methods used in shoot propagation of banana were .... forms of liquid medium application on the increase in dry matter in ... manual cutting, (2) Automation through use of liquid.

  6. Efficacy of cultural methods in the control of Rhizoctonia solani ...

    African Journals Online (AJOL)

    Rhizoctonia damping off of tomato caused by Rhizoctonia solani can be a ... of planting and influencing soil moisture levels either alone or in combination ... in combination with other cultural methods produced lower disease control and yield.

  7. A Kernel Time Structure Independent Component Analysis Method for Nonlinear Process Monitoring☆

    Institute of Scientific and Technical Information of China (English)

    Lianfang Cai; Xuemin Tian; Ni Zhang

    2014-01-01

    Kernel independent component analysis (KICA) is a newly emerging nonlinear process monitoring method, which can extract mutually independent latent variables cal ed independent components (ICs) from process var-iables. However, when more than one IC have Gaussian distribution, it cannot extract the IC feature effectively and thus its monitoring performance will be degraded drastical y. To solve such a problem, a kernel time struc-ture independent component analysis (KTSICA) method is proposed for monitoring nonlinear process in this paper. The original process data are mapped into a feature space nonlinearly and then the whitened data are calculated in the feature space by the kernel trick. Subsequently, a time structure independent component analysis algorithm, which has no requirement for the distribution of ICs, is proposed to extract the IC feature. Finally, two monitoring statistics are built to detect process faults. When some fault is detected, a nonlinear fault identification method is developed to identify fault variables based on sensitivity analysis. The proposed monitoring method is applied in the Tennessee Eastman benchmark process. Applications demonstrate the superiority of KTSICA over KICA.

  8. Intercultural and Cross-Cultural Communication Research: Some Reflections about Culture and Qualitative Methods

    Directory of Open Access Journals (Sweden)

    Maria Assumpta Aneas

    2009-01-01

    Full Text Available This article attempts to offer a response, from a general perspective, to the question of how culture reveals itself in the application of qualitative research methods in intercultural communication. When we use the term "culture" it is important to bear in mind that culturally attributed social interaction processes are themselves the result of socially constructed processes. They are part of an individual-collective dialectic with multiple potential meanings, which are emergent and in constant reformulation from a wide variety of social and cultural perspectives. Much of the recent research in intercultural communication has been directed towards the study of these systems of culturally related meanings. The literature we review offers perspectives from a variety of disciplines and insights into the role of culture in communication processes. URN: urn:nbn:de:0114-fqs0901519

  9. Cultural self-construals of Mexican, Spanish, and Danish college students: Beyond independent and interdependent self

    DEFF Research Database (Denmark)

    Santamaría, Andrés; de la Mata, Manuel L.; Hansen, Tia G. B.;

    2010-01-01

    De sidste par tiår har tværkulturel forskning vist vigtige forskelle i selvbegribelse. En dybere forståelse forudsætter imidlertid inddragelse af flere kulturer. Modsat Markus og Kitayamas distinktion mellem independent og interdependent selv, har forfattere som Kagitcibasi m.fl. argumenteret for...

  10. The structure of tissue on cell culture-extracted thyroglobulin is independent of its iodine content.

    Science.gov (United States)

    Delain, E; Aouani, A; Vignal, A; Couture-Tosi, E; Hovsépian, S; Fayet, G

    1987-02-01

    The major protein synthesized in vitro by the ovine thyroid cell line OVNIS 6H is the prothyroid hormone thyroglobulin. Purified from serum-free cell culture media using sucrose gradient centrifugation, the thyroglobulin dimer was analysed for iodine content and observed by electron microscopy. In their usual medium, the OVNIS 6H cells produce a very poorly iodinated thyroglobulin containing 0.05 I atom per molecule. When cultured with methimazole or propylthiouracil, two inhibitors of iodide organification, less than 0.007 I atom/molecules was found. These molecules purified from cell cultures were compared to those purified from ovine thyroid tissue containing 26 I atoms/mol. Despite large differences in iodine content, the three preparations all consist of 19 S thyroglobulin dimers with the classical ovoidal shape. The variability in size measurements remains in a 2% range for all thyroglobulin types. Consequently, no real significant variation can be found between the highly iodinated thyroglobulin isolated from tissue, and the poorly or non-iodinated thyroglobulins isolated from cells cultured with or without methimazole or propylthiouracil.

  11. Niche-independent high-purity cultures of Lgr5+ intestinal stem cells and their progeny

    NARCIS (Netherlands)

    Yin, Xiaolei; Farin, Henner F; van Es, Johan H; Clevers, Hans; Langer, Robert; Karp, Jeffrey M

    2014-01-01

    Although Lgr5(+) intestinal stem cells have been expanded in vitro as organoids, homogeneous culture of these cells has not been possible thus far. Here we show that two small molecules, CHIR99021 and valproic acid, synergistically maintain self-renewal of mouse Lgr5(+) intestinal stem cells, result

  12. Public Communication of Science and Cultural Consumption. Scientific Information as an Independent Element

    Directory of Open Access Journals (Sweden)

    Patricia Castellanos Pineda

    2010-11-01

    Full Text Available Nowadays, governments, universities, research centers and other institutions, like museums, invest its resources, human and financial, in public understanding of science. However, the abyss and indifference towards science seem not decrease or doing it slowly. This article looks at theoretical framework of scientific consumption from cultural consumption perspective and discusses some examples of it.

  13. Feeder-independent continuous culture of the PICM-19 pig liver stem cell line

    Science.gov (United States)

    The PICM-19 pig liver stem cell line is a bipotent cell line, i.e., capable of forming either bile ductules or hepatocyte monolayers in vitro, that was derived from the primary culture of pig embryonic stem cells. The cell line has been strictly feeder-dependent in that cell replication morphology,...

  14. Investigating the diversity of Pseudomonas spp. in soil using culture dependent and independent techniques

    DEFF Research Database (Denmark)

    Li, Lili; Al-Soud, Waleed Abu; Bermark, Lasse

    2013-01-01

    Less than 1% of bacterial populations present in environmental samples are culturable, meaning that cultivation will lead to an underestimation of total cell counts and total diversity. However, it is less clear whether this is also true for specific well-defined groups of bacteria for which sele...

  15. Revisiting bovine pyometra--new insights into the disease using a culture-independent deep sequencing approach.

    Science.gov (United States)

    Knudsen, Lif Rødtness Vesterby; Karstrup, Cecilia Christensen; Pedersen, Hanne Gervi; Agerholm, Jørgen Steen; Jensen, Tim Kåre; Klitgaard, Kirstine

    2015-02-25

    The bacteria present in the uterus during pyometra have previously been studied using bacteriological culturing. These studies identified Fusobacterium necrophorum and Trueperella pyogenes as the major contributors to the pathogenesis of pyometra. However, an increasing number of culture-independent studies have demonstrated that the bacterial diversity in most environments is underestimated in culture-based studies. Consequently, fastidious pyometra-associated pathogens may have been overlooked. Therefore, the primary purpose of this study was to investigate the diversity of bacteria in the uterus of cows with pyometra by using culture-independent 16S rRNA PCR combined with next generation sequencing. We investigated the microbial composition in the uterus of 21 cows with pyometra, which were obtained from a Danish slaughterhouse. Similar to the observations from the culture studies, Fusobacteriaceae, the family that F. necrophorum belongs to, was the operational taxonomic unit (OTU) observed in the largest quantities. By contrast, the Actinomycetaceae family, which includes T. pyogenes, constituted only 1% of the total number of reads. Thus we cannot confirm the previously reported role of species from this family in the pathogenesis of pyometra. Finally, we identified a large number of sequences representing three families of Gram-negative bacteria in the pyometra samples: Porphyromonadaceae, Mycoplasmataceae, and Pasteurellaceae. It is likely that these families comprise potential pathogenic species of a fastidious nature, which have been overlooked in previous studies. Our results increase the knowledge of the complexity of the pyometra microbiota and suggest that pathogens in addition to F. necrophorum may be involved in the pathogenesis of pyometra.

  16. Culture-independent bacterial community analysis of the salty-fermented fish paste products of Thailand and Laos.

    Science.gov (United States)

    Marui, Junichiro; Boulom, Sayvisene; Panthavee, Wanchai; Momma, Mari; Kusumoto, Ken-Ichi; Nakahara, Kazuhiko; Saito, Masayoshi

    2015-01-01

    A bacterial community analysis, using a culture-independent method (polymerase chain reaction-denaturing gradient gel electrophoresis), detected 17 species of bacteria including species of the genera Tetragenococcus, Lactobacillus, Pediococcus, Weissella Halanaerobium, Clostridium, and Sphingomonas in a traditional salty-fermented fish paste known as pla-ra or pa-daek in Thailand and Laos, which is used as a storage-stable multi-purpose seasoning. The representative genus of lactic acid bacteria seemed to vary in the 10 products collected from Thailand and Laos. Tetragenococci were common in products from central Thailand and Vientiane in Laos which had salinities of not less than 11% and pH values ranging from 5.6 to 6.1. However, lactobacilli were common in products from northern Thailand which had the lowest salinities (8.3-8.6%) and pH values (4.5-4.8) of all the samples examined. Two Lactobacillus and one Tetragenococcus species were detected in one product from northeastern Thailand containing 10% salt. These results suggest that salinity in pla-ra/pa-daek is an important determinant of the representative genus of lactic acid bacteria such as, Tetragenococcus or Lactobacillus. Additionally, differences in the acidity between these two groups seemed to be related to the production of d-/l-lactic acid in the lactic acid bacteria in each product. This is the first study to report a correlation between bacterial community structure and taste components in pla-ra/pa-daek products from various regions. This scientific work on a traditional fermented food will be useful in helping local producers meet differing consumer preferences in various regions.

  17. Culture-independent genome sequencing of clinical samples reveals an unexpected heterogeneity of infections by Chlamydia pecorum.

    Science.gov (United States)

    Bachmann, Nathan L; Sullivan, Mitchell J; Jelocnik, Martina; Myers, Garry S A; Timms, Peter; Polkinghorne, Adam

    2015-05-01

    Chlamydia pecorum is an important global pathogen of livestock, and it is also a significant threat to the long-term survival of Australia's koala populations. This study employed a culture-independent DNA capture approach to sequence C. pecorum genomes directly from clinical swab samples collected from koalas with chlamydial disease as well as from sheep with arthritis and conjunctivitis. Investigations into single-nucleotide polymorphisms within each of the swab samples revealed that a portion of the reads in each sample belonged to separate C. pecorum strains, suggesting that all of the clinical samples analyzed contained mixed populations of genetically distinct C. pecorum isolates. This observation was independent of the anatomical site sampled and the host species. Using the genomes of strains identified in each of these samples, whole-genome phylogenetic analysis revealed that a clade containing a bovine and a koala isolate is distinct from other clades comprised of livestock or koala C. pecorum strains. Providing additional evidence to support exposure of koalas to Australian livestock strains, two minor strains assembled from the koala swab samples clustered with livestock strains rather than koala strains. Culture-independent probe-based genome capture and sequencing of clinical samples provides the strongest evidence yet to suggest that naturally occurring chlamydial infections are comprised of multiple genetically distinct strains.

  18. Assessment of the microbial diversity at the surface of Livarot cheese using culture-dependent and independent approaches.

    Science.gov (United States)

    Mounier, J; Monnet, C; Jacques, N; Antoinette, A; Irlinger, F

    2009-07-31

    The microbial diversity of the surface of a commercial red-smear cheese, Livarot cheese, sold on the retail market was studied using culture-dependent and independent approaches. Forty yeasts and 40 bacteria from the cheese surface were collected, dereplicated using single-strand conformation polymorphism (SSCP) analysis and identified using rRNA gene sequencing for the culture-dependent approach. The culture-independent approach involved cloning and sequencing of the 16S rRNA gene and SSCP analysis from total DNA extracted from the cheese. The most dominant bacteria were Microbacterium gubbeenense, Leucobacter komagatae and Gram-negative bacteria from the Gamma-Proteobacteria class. Fluorescence in situ hybridization (FISH) analysis was also used to study the cheese microbial diversity with class-level and specific rRNA-targeted probes for bacteria and yeasts, respectively. FISH analysis confirmed that Gamma-Proteobacteria were important microorganisms in this cheese. Four specific FISH probes targeting the dominant yeasts present in the cheese, Candida catenulata, Candida intermedia, Geotrichum spp. and Yarrowia lipolytica, were also designed and evaluated. These probes allowed the detection of these yeasts directly in cheese. The use of the rRNA gene-based approach combined with FISH analysis was useful to investigate the diversity of a surface microbial consortium from cheese.

  19. Studying generalised dark matter interactions with extended halo-independent methods

    CERN Document Server

    Kahlhoefer, Felix

    2016-01-01

    The interpretation of dark matter direct detection experiments is complicated by the fact that neither the astrophysical distribution of dark matter nor the properties of its particle physics interactions with nuclei are known in detail. To address both of these issues in a very general way we develop a new framework that combines the full formalism of non-relativistic effective interactions with state-of-the-art halo-independent methods. This approach makes it possible to analyse direct detection experiments for arbitrary dark matter interactions and quantify the goodness-of-fit independent of astrophysical uncertainties. We employ this method in order to demonstrate that the degeneracy between astrophysical uncertainties and particle physics unknowns is not complete. Certain models can be distinguished in a halo-independent way using a single ton-scale experiment based on liquid xenon, while other models are indistinguishable with a single experiment but can be separated using combined information from seve...

  20. Studying generalised dark matter interactions with extended halo-independent methods

    Energy Technology Data Exchange (ETDEWEB)

    Kahlhoefer, Felix [DESY, Notkestraße 85,D-22607 Hamburg (Germany); Wild, Sebastian [Physik-Department T30d, Technische Universität München,James-Franck-Straße 1, D-85748 Garching (Germany)

    2016-10-20

    The interpretation of dark matter direct detection experiments is complicated by the fact that neither the astrophysical distribution of dark matter nor the properties of its particle physics interactions with nuclei are known in detail. To address both of these issues in a very general way we develop a new framework that combines the full formalism of non-relativistic effective interactions with state-of-the-art halo-independent methods. This approach makes it possible to analyse direct detection experiments for arbitrary dark matter interactions and quantify the goodness-of-fit independent of astrophysical uncertainties. We employ this method in order to demonstrate that the degeneracy between astrophysical uncertainties and particle physics unknowns is not complete. Certain models can be distinguished in a halo-independent way using a single ton-scale experiment based on liquid xenon, while other models are indistinguishable with a single experiment but can be separated using combined information from several target elements.

  1. GNSS Vertical Coordinate Time Series Analysis Using Single-Channel Independent Component Analysis Method

    Science.gov (United States)

    Peng, Wei; Dai, Wujiao; Santerre, Rock; Cai, Changsheng; Kuang, Cuilin

    2017-02-01

    Daily vertical coordinate time series of Global Navigation Satellite System (GNSS) stations usually contains tectonic and non-tectonic deformation signals, residual atmospheric delay signals, measurement noise, etc. In geophysical studies, it is very important to separate various geophysical signals from the GNSS time series to truthfully reflect the effect of mass loadings on crustal deformation. Based on the independence of mass loadings, we combine the Ensemble Empirical Mode Decomposition (EEMD) with the Phase Space Reconstruction-based Independent Component Analysis (PSR-ICA) method to analyze the vertical time series of GNSS reference stations. In the simulation experiment, the seasonal non-tectonic signal is simulated by the sum of the correction of atmospheric mass loading and soil moisture mass loading. The simulated seasonal non-tectonic signal can be separated into two independent signals using the PSR-ICA method, which strongly correlated with atmospheric mass loading and soil moisture mass loading, respectively. Likewise, in the analysis of the vertical time series of GNSS reference stations of Crustal Movement Observation Network of China (CMONOC), similar results have been obtained using the combined EEMD and PSR-ICA method. All these results indicate that the EEMD and PSR-ICA method can effectively separate the independent atmospheric and soil moisture mass loading signals and illustrate the significant cause of the seasonal variation of GNSS vertical time series in the mainland of China.

  2. Method of independent timesteps in the numerical solution of initial value problems

    Energy Technology Data Exchange (ETDEWEB)

    Porter, A.P.

    1976-07-21

    In the numerical solution of initial-value problems in several independent variables, the timestep is controlled, especially in the presence of shocks, by a small portion of the logical mesh, what one may call the crisis zone. One is frustrated by the necessity of doing in the whole mesh frequent calculations required by only a small part of the mesh. It is shown that it is possible to choose different timesteps natural to different parts of the mesh and to advance each zone in time only as often as is appropriate to that zone's own natural timestep. Prior work is reviewed and for the first time an investigation of the conditions for well-posedness, consistency and stability in independent timesteps is presented; a new method results. The prochronic and parachronic Cauchy surfaces are identified; and the reasons (well-posedness) for constraining the Cauchy surfaces to be prochronic (as distinct from the method of Grandey), that is, to lie prior to the time of the crisis zone (the zone of least timestep), are indicated. Stability (in the maximum norm) of parabolic equations and (in the L2 norm) of hyperbolic equations is reviewed, without restricting the treatment to linear equations or constant coefficients, and stability of the new method is proven in this framework. The details of the method of independent timesteps, the rules for choosing timesteps and for deciding when to update and when to skip zones, and the method of joining adjacent regions of differing timestep are described. The stability of independent timestep difference schemes is analyzed and exhibited. The economic advantages of the method, which often amount to an order-of-magnitude decrease in running time relative to conventional or implicit difference methods, are noted.

  3. A new process monitoring method based on noisy time structure independent component analysis

    Institute of Scientific and Technical Information of China (English)

    Lianfang Cai; Xuemin Tian

    2015-01-01

    Conventional process monitoring method based on fast independent component analysis (FastICA) cannot take the ubiquitous measurement noises into account and may exhibit degraded monitoring performance under the adverse effects of the measurement noises. In this paper, a new process monitoring approach based on noisy time structure ICA (NoisyTSICA) is proposed to solve such problem. A NoisyTSICA algorithm which can consider the measurement noises explicitly is firstly developed to estimate the mixing matrix and extract the independent components (ICs). Subsequently, a monitoring statistic is built to detect process faults on the basis of the recur-sive kurtosis estimations of the dominant ICs. Lastly, a contribution plot for the monitoring statistic is constructed to identify the fault variables based on the sensitivity analysis. Simulation studies on the continuous stirred tank reactor system demonstrate that the proposed NoisyTSICA-based monitoring method outperforms the conven-tional FastICA-based monitoring method.

  4. A Quick Method for Metarhizium anisopliae Isolation from Cultural Soils

    Directory of Open Access Journals (Sweden)

    M. A. Tajick Ghanbary

    2009-01-01

    Full Text Available Problem statement: Fungi are one of the most active members in biological community of cultural soils. Many saprophyte and facultative parasitic fungi live in soil. Metarhizium anisopliae, one of the most famous soil inhabitant entomopathogens has a virulence potential on plant and animal pests. Approach: Introducing a new method for its isolation from soil was an applied method to find it without any limitation. Metarhizium anisopliae shifts to saprophytic phase and remain alive within soil in absence of susceptible host. As a shortcut, we can transfer the fungus from soil to lab by culturing soil suspension. One hundred cultural soil samples from different regions of Iran were tested to finding Metarhizium isolates. Culturing 1:5000-1:10000 soil suspension on artificial medium containing necessary macro and micronutrients for fungal growth were resulted in isolation. Metarhizium anisopliae isolates were harvested seven days after culturing the suspensions. All isolates were inoculated in 50 mL PDB in destruxin production assay and 7 days later broth medium was filtrated by using filter paper. Culture filtrates were extracted and in bioassays they were sprayed on larva of citrus leaf miner. Results: Nine isolates of Metarhizium anisopliae were harvested. Microscopic studies showed that morphological features had complete coincidence with valid descriptions of the fungus. Bioassay confirmed that all harvested isolates secrete active and effective destruxin in broth. Conclusion: Isolation of Metarhizium by culturing the soil suspension, a useful method for more studies of the entomopathogen at different geographical regions. Native populations of this fungus had special importance in local biological control programs. This procedure was a costs- and time-effective method for pathogen isolation.

  5. Estimation of effective lens position using a method independent of preoperative keratometry readings.

    LENUS (Irish Health Repository)

    Dooley, Ian

    2012-02-01

    PURPOSE: To evaluate the validity of a keratometry (K)-independent method of estimating effective lens position (ELP) before phacoemulsification cataract surgery. SETTING: Institute of Eye Surgery, Whitfield Clinic, Waterford, Ireland. DESIGN: Evaluation of diagnostic test or technology. METHODS: The anterior chamber diameter and corneal height in eyes scheduled for cataract surgery were measured with a rotating Scheimpflug camera. Corneal height and anterior chamber diameter were used to estimate the ELP in a K-independent method (using the SRK\\/T [ELP(rs)] and Holladay 1 [ELP(rh)] formulas). RESULTS: The mean ELP was calculated using the traditional (mean ELP(s) 5.59 mm +\\/- 0.52 mm [SD]; mean ELP(h) 5.63 +\\/- 0.42 mm) and K-independent (mean ELP(rs) 5.55 +\\/- 0.42 mm; mean ELP(rh) +\\/- SD 5.60 +\\/- 0.36 mm) methods. Agreement between ELP(s) and ELP(rs) and between ELP(h) and ELP(rh) were represented by Bland-Altman plots, with mean differences (+\\/- 1.96 SD) of 0.06 +\\/- 0.65 mm (range -0.59 to +0.71 mm; P=.08) in association with ELP(rs) and -0.04 +\\/- 0.39 mm (range -0.43 to +0.35 mm; P=.08) in association with ELP(rh). The mean absolute error for ELP(s) versus ELP(rs) estimation and for ELP(h) versus ELP(rh) estimation was 0.242 +\\/- 0.222 mm (range 0.001 to 1.272 mm) and 0.152 +\\/- 0.137 mm (range 0.001 to 0.814 mm), respectively. CONCLUSION: This study confirms that the K-independent ELP estimation method is comparable to traditional K-dependent methods and may be useful in post-refractive surgery patients.

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

    Science.gov (United States)

    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), mode-M had significantly higher shrimp yield, survival rate and feed conversion rate ( P vannamei.

  7. Independent Study: The Culture of Mentored Undergraduate Research at The College of Wooster (Invited)

    Science.gov (United States)

    Pollock, M.; Judge, S.; Wiles, G. C.; Wilson, M. A.

    2013-12-01

    The foundation of a Wooster education is the Independent Study (I.S.) program. Established in 1947, the I.S. program is widely recognized as an exemplary undergraduate research experience (AAC&U; US News and World Report; College that Change Lives by Loren Pope). I.S. requires every Wooster student to complete an original research project. This presentation describes the details of the Wooster I.S. and, based on our experiences, gives strategies for a successful mentored undergraduate research program. Overall, the I.S. process resembles a graduate research project. Students typically begin their work in the spring of their junior year when they review the literature, learn techniques, and write a proposal for their Senior I.S. research. Many students conduct field and lab work over the following summer, although this is not a requirement of the program. In their senior year, students work one-on-one with faculty members and sometimes in small (~4 person) research groups to drive their projects forward with an increasing sense of independence. I.S. culminates in a written thesis and oral defense. Most of our students present their work at national meetings and many projects are published in peer-reviewed journals. The success of the I.S. program is largely the result of two key components: (1) the integration of undergraduate research into the curriculum, and (2) the focus on student mentoring. We have thoughtfully structured our courses so that, as students move toward I.S., they progress from concrete to abstract concepts, and from simple to complex skills. The College also recognizes the value of I.S by assigning it credit; Students earn a full course credit for each semester of I.S. (3 courses total) and there is some credit in the faculty teaching load for I.S. advising. Advisors are really mentors who are invested in their students' academic and scholarly success. As mentors, we emphasize collaboration, provide guidance and support, and hold students

  8. Improved Cell Culture Method for Growing Contracting Skeletal Muscle Models

    Science.gov (United States)

    Marquette, Michele L.; Sognier, Marguerite A.

    2013-01-01

    An improved method for culturing immature muscle cells (myoblasts) into a mature skeletal muscle overcomes some of the notable limitations of prior culture methods. The development of the method is a major advance in tissue engineering in that, for the first time, a cell-based model spontaneously fuses and differentiates into masses of highly aligned, contracting myotubes. This method enables (1) the construction of improved two-dimensional (monolayer) skeletal muscle test beds; (2) development of contracting three-dimensional tissue models; and (3) improved transplantable tissues for biomedical and regenerative medicine applications. With adaptation, this method also offers potential application for production of other tissue types (i.e., bone and cardiac) from corresponding precursor cells.

  9. Monitoring of the microbiota of fermented sausages by culture independent rRNA-based approaches.

    Science.gov (United States)

    Greppi, Anna; Ferrocino, Ilario; La Storia, Antonietta; Rantsiou, Kalliopi; Ercolini, Danilo; Cocolin, Luca

    2015-11-06

    In Italy, fermented sausages (called "salami") are consumed in large quantities. Salami samples from a local meat factory in the area of Torino were analyzed at 0, 3, 7, 30 and 45 days of ripening. Swab samples from the production environment were also collected at the beginning of the experiment. The diversity of metabolically active microbiota occurring during the natural fermentation of salami was evaluated by using RT-PCR-DGGE coupled with RNA-based pyrosequencing of the 16S rRNA gene. A culture-dependent approach was also applied to identify and characterize isolated Staphylococcaceae and LAB populations. Staphylococcus succinus, Staphylococcus xylosus and Lactobacillus sakei were the species most frequently isolated during the maturation time. Rep-PCR analysis showed that S. succinus and S. xylosus isolated from swabs and salami samples clustered together, suggesting possible contamination during the production process. RT-PCR-DGGE and rRNA-based pyrosequencing showed that the metabolically active populations were dominated by S. succinus, Lb. sakei and Leuconostoc carnosum. In this specific case study, only a few species belonging to Staphylococcaceae, Lactobacillaceae and Leuconostocaceae may be metabolically active and contribute to determine the final characteristics of the products.

  10. A new and simple calibration-independent method for measuring the beam energy of a cyclotron.

    Science.gov (United States)

    Gagnon, Katherine; Jensen, Mikael; Thisgaard, Helge; Publicover, Julia; Lapi, Suzanne; McQuarrie, Steve A; Ruth, Thomas J

    2011-01-01

    This work recommends a new and simple-to-perform method for measuring the beam energy of an accelerator. The proposed method requires the irradiation of two monitor foils interspaced by an energy degrader. The primary advantage of the proposed method, which makes this method unique from previous energy evaluation strategies that employ the use of monitor foils, is that this method is independent of the detector efficiency calibration. This method was evaluated by performing proton activation of (nat)Cu foils using both a cyclotron and a tandem Van de Graaff accelerator. The monitor foil activities were read using a dose calibrator set to an arbitrary calibration setting. Excellent agreement was noted between the nominal and measured proton energies.

  11. A new and simple calibration-independent method for measuring the beam energy of a cyclotron

    Energy Technology Data Exchange (ETDEWEB)

    Gagnon, Katherine, E-mail: kgagnon1@ualberta.c [Cross Cancer Institute, Edmonton PET Centre, University of Alberta, Edmonton, AB, T6G 1Z2 (Canada); Jensen, Mikael; Thisgaard, Helge [Hevesy Laboratory, Risoe-DTU, Technical University of Denmark, Frederiksborgvej 399, DK-4000 Roskilde (Denmark); Publicover, Julia; Lapi, Suzanne [TRIUMF, 4004 Wesbrook Mall, Vancouver, BC, V6T 2A3 (Canada); McQuarrie, Steve A. [Cross Cancer Institute, Edmonton PET Centre, University of Alberta, Edmonton, AB, T6G 1Z2 (Canada); Ruth, Thomas J. [TRIUMF, 4004 Wesbrook Mall, Vancouver, BC, V6T 2A3 (Canada)

    2011-01-15

    This work recommends a new and simple-to-perform method for measuring the beam energy of an accelerator. The proposed method requires the irradiation of two monitor foils interspaced by an energy degrader. The primary advantage of the proposed method, which makes this method unique from previous energy evaluation strategies that employ the use of monitor foils, is that this method is independent of the detector efficiency calibration. This method was evaluated by performing proton activation of {sup nat}Cu foils using both a cyclotron and a tandem Van de Graaff accelerator. The monitor foil activities were read using a dose calibrator set to an arbitrary calibration setting. Excellent agreement was noted between the nominal and measured proton energies.

  12. Method-independent, Computationally Frugal Convergence Testing for Sensitivity Analysis Techniques

    Science.gov (United States)

    Mai, Juliane; Tolson, Bryan

    2017-04-01

    The increasing complexity and runtime of environmental models lead to the current situation that the calibration of all model parameters or the estimation of all of their uncertainty is often computationally infeasible. Hence, techniques to determine the sensitivity of model parameters are used to identify most important parameters or model processes. All subsequent model calibrations or uncertainty estimation procedures focus then only on these subsets of parameters and are hence less computational demanding. While the examination of the convergence of calibration and uncertainty methods is state-of-the-art, the convergence of the sensitivity methods is usually not checked. If any, bootstrapping of the sensitivity results is used to determine the reliability of the estimated indexes. Bootstrapping, however, might as well become computationally expensive in case of large model outputs and a high number of bootstraps. We, therefore, present a Model Variable Augmentation (MVA) approach to check the convergence of sensitivity indexes without performing any additional model run. This technique is method- and model-independent. It can be applied either during the sensitivity analysis (SA) or afterwards. The latter case enables the checking of already processed sensitivity indexes. To demonstrate the method independency of the convergence testing method, we applied it to three widely used, global SA methods: the screening method known as Morris method or Elementary Effects (Morris 1991, Campolongo et al., 2000), the variance-based Sobol' method (Solbol' 1993, Saltelli et al. 2010) and a derivative-based method known as Parameter Importance index (Goehler et al. 2013). The new convergence testing method is first scrutinized using 12 analytical benchmark functions (Cuntz & Mai et al. 2015) where the true indexes of aforementioned three methods are known. This proof of principle shows that the method reliably determines the uncertainty of the SA results when different

  13. The Ecophysiologies of Thiomargarita spp. Interrogated Using Culture-independent Approaches

    Science.gov (United States)

    Bailey, J.; Flood, B.; Ricci, E.; Jones, D. S.; Delherbe, N.

    2016-12-01

    Bacteria of the family Beggiatoaceae are primarily chemolithotrophs or mixotrophs that live at interfaces between nitrate or oxygen, and hydrogen sulfide. Thiomargarita spp., the largest of the sulfur bacteria, have yet to be cultivated and their ecologies and physiologies have yet to be fully elucidated. Despite our current inability to sustain Thiomargarita isolates in the lab, their large size allows for individual cells to be followed in time-course experiments, or to be individually targeted for `omics-based investigations. Our metagenomic investigations of Thiomargarita nelsonii identified genes involved in polyphosphate metabolism in the Thiomargarita genome, as well as genes involved in coupling sulfide oxidation to both denitrification and nitrate-reducing ammonification. Our results also show that T. nelsonii may also use non-canonical substrates such as As and H2. Our genomic results were complemented by the use of a tetrazolium-based dye that measures the flux of NADH production from catabolic pathways via a colorimetric response. Staining with this dye allows for metabolism to be detected, even in the absence of observable cell division. Redox staining suggests that respiratory activity occurs primarily under anoxic conditions. We are also in the process of employing fluorescently-labeled amino acids to monitor the synthesis of peptidoglycan, in order to estimate cell growth rates that may be very slow. Finally, we are investigating interactions between Thiomargarita and their epibiont bacterial communities. Symbiotic interactions with attached bacteria may be critical to the ecophysiology of Thiomargarita. Through these cultivation-independent approaches, we hope to better understand Thiomargarita spp. and perhaps gain insights into why they have thus far resisted efforts to cultivate them in the laboratory.

  14. Sponge cell culture? A molecular identification method for sponge cells

    NARCIS (Netherlands)

    Sipkema, D.; Heilig, G.H.J.; Akkermans, A.D.L.; Osinga, R.; Tramper, J.; Wijffels, R.H.

    2003-01-01

    Dissociated sponge cells are easily confused with unicellular organisms. This has been an obstacle in the development of sponge-cell lines. We developed a molecular detection method to identify cells of the sponge Dysidea avara in dissociated cell cultures. The 18S ribosomal RNA gene from a Dysidea

  15. Salesperson Self-Regulation of Pride: Effects on Adaptability, Effort, and Citizenship Behaviors between Independent-Based and Interdependent-Based Cultures

    NARCIS (Netherlands)

    Bagozzi, R.P.; Belschak, F.; Verbeke, W.; Gavino Jr., J.R.

    2016-01-01

    We investigate and compare how salespersons within an independent-based culture (the Netherlands) and an interdependent-based culture (the Philippines) experience and self-regulate pride that is evoked through praise and recognition by their managers. This self-regulation differentially influences b

  16. Towards nonaxisymmetry; initial results using the Flux Coordinate Independent method in BOUT++

    CERN Document Server

    Shanahan, Brendan; Dudson, Ben

    2016-01-01

    Fluid simulation of stellarator edge transport is difficult due to the complexities of mesh generation; the stochastic edge and strong nonaxisymmetry inhibit the use of field aligned coordinate systems. The recent implementation of the Flux Coordinate Independent method for calculating parallel derivatives in BOUT++ has allowed for more complex geometries. Here we present initial results of nonaxisymmetric diffusion modelling as a step towards stellarator turbulence modelling. We then present initial (non-turbulent) transport modelling using the FCI method and compare the results with analytical calculations. The prospects for future stellarator transport and turbulence modelling are discussed.

  17. Reliable transgene-independent method for determining Sleeping Beauty transposon copy numbers

    Directory of Open Access Journals (Sweden)

    Kolacsek Orsolya

    2011-03-01

    Full Text Available Abstract Background The transposon-based gene delivery technique is emerging as a method of choice for gene therapy. The Sleeping Beauty (SB system has become one of the most favored methods, because of its efficiency and its random integration profile. Copy-number determination of the delivered transgene is a crucial task, but a universal method for measuring this is lacking. In this paper, we show that a real-time quantitative PCR-based, transgene-independent (qPCR-TI method is able to determine SB transposon copy numbers regardless of the genetic cargo. Results We designed a specific PCR assay to amplify the left inverted repeat-direct repeat region of SB, and used it together with the single-copy control gene RPPH1 and a reference genomic DNA of known copy number. The qPCR-TI method allowed rapid and accurate determination of SB transposon copy numbers in various cell types, including human embryonic stem cells. We also found that this sensitive, rapid, highly reproducible and non-radioactive method is just as accurate and reliable as the widely used blotting techniques or the transposon display method. Because the assay is specific for the inverted repeat region of the transposon, it could be used in any system where the SB transposon is the genetic vehicle. Conclusions We have developed a transgene-independent method to determine copy numbers of transgenes delivered by the SB transposon system. The technique is based on a quantitative real-time PCR detection method, offering a sensitive, non-radioactive, rapid and accurate approach, which has a potential to be used for gene therapy.

  18. Dispatch Method for Independently Owned Hydropower Plants in the Same River Flow

    Directory of Open Access Journals (Sweden)

    Slavko Krajcar

    2012-09-01

    Full Text Available This paper proposes a coexistence model for two independent companies both operating hydropower plants in the same river flow, based on a case study of the Cetina river basin in Croatia. Companies are participants of the day-ahead electricity market. The incumbent company owns the existing hydropower plants and holds concessions for the water. The new company decides to build a pump storage hydropower plant that uses one of the existing reservoirs as its lower reservoir. Meeting reservoir water balance is affected by decisions by both companies which are independently seeking maximal profit. Methods for water use settlement and preventing of spillage are proposed. A mixed-integer linear programming approach is used. Head effects on output power levels are also considered. Existences of dispatches that satisfy both companies are shown.

  19. High-throughput sequencing and metagenomics: moving forward in the culture-independent analysis of food microbial ecology.

    Science.gov (United States)

    Ercolini, Danilo

    2013-05-01

    Following recent trends in environmental microbiology, food microbiology has benefited from the advances in molecular biology and adopted novel strategies to detect, identify, and monitor microbes in food. An in-depth study of the microbial diversity in food can now be achieved by using high-throughput sequencing (HTS) approaches after direct nucleic acid extraction from the sample to be studied. In this review, the workflow of applying culture-independent HTS to food matrices is described. The current scenario and future perspectives of HTS uses to study food microbiota are presented, and the decision-making process leading to the best choice of working conditions to fulfill the specific needs of food research is described.

  20. PURE CULTURE METHOD: GIARDIA LAMBLIA FROM DIFFERENT STOOL SAMPLES

    Directory of Open Access Journals (Sweden)

    H.A YOUSEFI

    2000-03-01

    Full Text Available Introduction. Giardiasis is one of the health problems in the world including Iran. To determine the biochemical and biological problems and also identification of various strains, it is essential to obtain pure culture and then mass production of Giardia lamblia. The goal of this study was to isolate this protozoa purely.
    Methods. Giardia lamblia cysts were isolated from 50 stool samples by use of floating of a four - layer of sucrose method. The cysts were transfered to an inducing solution. Subsequently, they were cultured in a modified culture medium (TYIS-33. Following excystation of trophozoite and its multiplication, the parasite was caltured and purified.
    Findings. Excitation of trophozoite was observed in 40 samples (80 percent from which 22 samples (55 percent yielded pure culture. The doubling time was approximately 13hr and the peak of parasite was observed between third and fourth days.
    Conclusion. The proliferation and growth rate of Giardia lamblia have enabled us to use this method widely. Cystein and ascorbic acid which are present in the induction solution, have a key role in excystation of trophozoite. Purification and passage of samples has facilitated the culture of this parasite in vitro. Therefore this method has yielded better results in comparison with other studies. This is probably due to a decrease in the amount of bovine bile or using different strains of Giardia lamblia in the present study.

  1. Cytobrush-culture method to diagnose tinea capitis.

    Science.gov (United States)

    Bonifaz, Alexandro; Isa-Isa, Rafael; Araiza, Javier; Cruz, Cecilia; Hernández, Marco A; Ponce, Rosa Maria

    2007-06-01

    This is a comparative study to isolate the dermatophytes of tinea capitis using the cytobrush and comparing it versus the standard method. A prospective, observational, comparative trial of 178 probable cases of tinea capitis was conducted in two dermatological centers. Each patient underwent mycological tests that included direct exam with KOH and cultures with either of two methods: scraping the scalp to remove hair and cell debris, and the cytobrush. A total of 135 clinically and mycologically proven cases of tinea capitis were included; 119 were non-inflammatory and 16 inflammatory tinea. A total of 131 had a positive direct exam and subsequent primary isolation cultures were obtained in 135 cases. The main dermatophytes isolated were Microsporum canis (68%) and Trichophyton tonsurans (20%). A total of 115/135 (85.1%), were detected with the traditional method, with an average of 11.2 days until positive, while the number detected with the cytobrush was 132/135 (97.7%) with an average of 8.5 days until positive. The chi-square statistical method showed that the cytobrush culture was superior to the standard one with a chi-square of 5.078 (P = 0.025), with a statistically significant difference versus the standard method.

  2. Genomic instability of human embryonic stem cell lines using different passaging culture methods.

    Science.gov (United States)

    Tosca, Lucie; Feraud, Olivier; Magniez, Aurélie; Bas, Cécile; Griscelli, Frank; Bennaceur-Griscelli, Annelise; Tachdjian, Gérard

    2015-01-01

    Human embryonic stem cells exhibit genomic instability that can be related to culture duration or to the passaging methods used for cell dissociation. In order to study the impact of cell dissociation techniques on human embryonic stem cells genomic instability, we cultured H1 and H9 human embryonic stem cells lines using mechanical/manual or enzymatic/collagenase-IV dissociation methods. Genomic instability was evaluated at early (p60) passages by using oligonucleotide based array-comparative genomic hybridization 105 K with a mean resolution of 50 Kb. DNA variations were mainly located on subtelomeric and pericentromeric regions with sizes <100 Kb. In this study, 9 recurrent genomic variations were acquired during culture including the well known duplication 20q11.21. When comparing cell dissociation methods, we found no significant differences between DNA variations number and size, DNA gain or DNA loss frequencies, homozygous loss frequencies and no significant difference on the content of genes involved in development, cell cycle tumorigenesis and syndrome disease. In addition, we have never found any malignant tissue in 4 different teratoma representative of the two independent stem cell lines. These results show that the occurrence of genomic instability in human embryonic stem cells is similar using mechanical or collagenase IV-based enzymatic cell culture dissociation methods. All the observed genomic variations have no impact on the development of malignancy.

  3. Responsiveness to PI3K and MEK inhibitors in breast cancer. Use of a 3D culture system to study pathways related to hormone independence in mice.

    Directory of Open Access Journals (Sweden)

    Maria Laura Polo

    Full Text Available BACKGROUND: A significant proportion of breast cancer patients face failure of endocrine therapy due to the acquisition of endocrine resistance. We have explored mechanisms involved in such disease progression by using a mouse breast cancer model that is induced by medroxyprogesterone acetate (MPA. These tumors transit through different stages of hormone sensitivity. However, when cells from tumor variants were seeded on plastic, all were stimulated by progestins and inhibited by antiprogestins such as RU486. Furthermore, cells from a RU486-resistant tumor variant recovered antiprogestin sensitivity. HYPOTHESIS: A three-dimensional (3D culture system, by maintaining differential cellular organization that is typical of each tumor variant, may allow for the maintenance of particular hormone responses and thus be appropriate for the study of the effects of specific inhibitors of signaling pathways associated with disease progression. METHOD: We compared the behavior of tumors growing in vivo and cancer cells ex vivo (in 3D Matrigel. In this system, we evaluated the effects of kinase inhibitors and hormone antagonists on tumor growth. PRINCIPAL FINDINGS: LY294002, a PI3K/AKT pathway inhibitor, decreased both tumor growth in vivo and cell survival in Matrigel in MPA-independent tumors with higher AKT activity. Induction of cell death by anti-hormones such as ICI182780 and ZK230211 was more effective in MPA-dependent tumors with lower AKT activity. Inhibition of MEK with PD98059 did not affect tumor growth in any tested variant. Finally, while Matrigel reproduced differential responsiveness of MPA-dependent and -independent breast cancer cells, it was not sufficient to preserve antiprogestin resistance of RU486-resistant tumors. CONCLUSION: We demonstrated that the PI3K/AKT pathway is relevant for MPA-independent tumor growth. Three-dimensional cultures were useful to test the effects of kinase inhibitors on breast cancer growth and highlight the

  4. The Effect of Three Culture Methods on Intensive Culture System of Pacific White Shrimp (Litopenaeus vannamei)

    Institute of Scientific and Technical Information of China (English)

    MA Zhen; WAN Rong; SONG Xiefa; GAO Lei

    2013-01-01

    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 84d.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 50d 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 ofL.vannamei.

  5. Study of Nosocomial Clostridium Difficile Diarrhea by Culture and Tissue Culture Methods

    Directory of Open Access Journals (Sweden)

    MH Salari

    2007-06-01

    Full Text Available Background: Clostridium difficult is an identified cause of antibiotic-associated diarrhea, antibiotic-associated colitis, pseudomembranous colitis and nosocomial diarrhea. The purpose of this study was to investigate the prevalence of nosocomial C. difficile-associated diarrhea in Tehran University of Medical Science Hospitals. Methods: In this study a total of 942 stool samples from patients with nosocomial diarrhea that were hospitalized in Imam Khomeini hospital, Shariati hospital and Children clinical center were collected. The samples were cultured on a selective cycloserine cefoxitin fructose agar (CCFA and incubated in anaerobic conditions, at 37 °C for 5 days. Isolated C. difficile by conventional biochemical tests, bacterial cytotoxicity by Vero tissue culture and antimicrobial sensitivity to antibiotics by Kirby Bauer method (disk diffusion were investigated. Results: Of the total patients, 57 Toxigenic C. difficile (6.1% were isolated. Results of statistical analysis show significant differences between the rate of isolated Toxigenic C. difficile and age group of patients (P< 0.05. Among the units of selected hospitals, Toxigenic C. difficile was isolated most frequently in gastroenterology of Children clinical center. Meanwhile, the isolated Toxigenic C. difficile were sensitive to vancomycin, Chloramphenicol and ceftriaxone. Conclusion: Our findings show that, Toxigenic C. difficile was found in 6.1% hospitalized patients. Therefore, further studies to evaluate the role of Toxigenic C. difficile in nosocomial diarrhea processes, ecological and pathogenic terms by culture, Tissue culture and molecular methodes are suggested.

  6. Culture-dependent and culture-independent analyses reveal no prokaryotic community shifts or recovery of Serratia marcescens in Acropora palmata with white pox disease.

    Science.gov (United States)

    Lesser, Michael P; Jarett, Jessica K

    2014-06-01

    Recently, the etiological agent of white pox (WP) disease, also known as acroporid serratiosis, in the endangered coral Acropora palmata is the enteric bacterium Serratia marcescens with the source being localized sewage release onto coastal coral reef communities. Here, we show that both culture-dependent and culture-independent approaches could not recover this bacterium from samples of tissue and mucus from A. palmata colonies affected by WP disease in the Bahamas, or seawater collected adjacent to A. palmata colonies. Additionally, a metagenetic 16S rRNA pyrosequencing study shows no significant difference in the bacterial communities of coral tissues with and without WP lesions. As recent studies have shown for other coral diseases, S. marcescens cannot be identified in all cases of WP disease in several geographically separated populations of A. palmata with the same set of signs. As a result, its identification as the etiological agent of WP disease, and cause of a reverse zoonosis, cannot be broadly supported. However, the prevalence of WP disease associated with S. marcescens does appear to be associated with proximity to population centers, and research efforts should be broadened to examine this association, and to identify other causes of this syndrome.

  7. Ethnic and Cultural Aspects in the Development of Kazakh Theatres during the Independence Period: The Problems of Human Existence

    Directory of Open Access Journals (Sweden)

    Askhat Mayemirov

    2015-12-01

    Full Text Available The article gives a detailed description of folklore motifs that became examples for performances in the Kazakh theatre. The interrelation of myth and reality, used as the nature of time, is represented by an existential communicational human existence in the ethno-cultural discourse. Here the conditions of existence for the heroes of both the reality and myth are the same. The comparison of the heroes’ lifestyle with Kazakh folklore gives a new interpretation quality to the director’s conception and dramaturgy. The heroes’ destinies under the totalitarian era of Stalin, Khrushchev, and Brezhnev are imbued with personification of folklore myths and legends. The examples given in the study are related to the cultural-philosophical paradigms of modernity and the values of current ethno-cultural transformations. Thus, the inclusion of folklore into the theatre becomes an influential art trend, in which art seeks to give a comprehensive and vivid real-life picture of a person who fights for his/her own destiny and justice. The Gabit Musrepov Kazakh Academic Theatre for Children and Youth solves these problems, using the comparative method of folk stories, encouraging the viewer to understand and appreciate the philosophical meaning of human life and existence. The study uses the ratio of diachronic and synchronic presentation of texts and artefacts as regards current events from the perspective of folklore studies, theatre studies, and art history, as well as from the cultural-philosophical viewpoint.

  8. From Cultural Path to Cultural Route: a Value-Led Risk Management Method for via Iulia Augusta in Albenga (italy)

    Science.gov (United States)

    Van Meerbeek, L.; Barazzetti, L.; Valente, R.

    2017-08-01

    Today, the field of cultural heritage faces many challenges: cultural heritage is always at risk, the large amount of heritage information is often fragmented, climate change impacts cultural heritage and heritage recording can be time-consuming and often results in low accuracy. Four objectives, related to the challenges, were defined during this research work. It proposes a relevant value-led risk management method for cultural heritage, it identifies climate change impact on cultural heritage, it suggests a database lay-out for cultural heritage and demonstrates the potential of remote sensing tools for cultural heritage. The Via Iulia Augusta, a former Roman road in Albenga, was used as case study.

  9. Numerical methods for 3D tokamak simulations using a flux-surface independent grid

    Energy Technology Data Exchange (ETDEWEB)

    Stegmeir, A.; Coster, D.; Maj, O.; Lackner, K. [Max-Planck-Institut fuer Plasmaphysik, EURATOM Association, 85748 Garching (Germany)

    2014-06-15

    A numerical approach for 3D Tokamak simulations using a flux surface independent grid is presented. The grid consists of few poloidal planes with a Cartesian isotropic grid within each poloidal plane. Perpendicular operators can be discretised within a poloidal plane using standard second order finite difference methods. The discretisation of parallel operators is achieved with a field line following map and an interpolation. The application of the support operator method to the parallel diffusion operator conserves the self-adjointness of the operator on the discrete level and keeps the numerical decay rate at a low level. The developed numerical methods can be applied to geometries where an X-point is present. (copyright 2014 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  10. Transformation-based spherical cloaks designed by an implicit transformation-independent method: theory and optimization

    Energy Technology Data Exchange (ETDEWEB)

    Novitsky, Andrey [Department of Theoretical Physics, Belarusian State University, Nezavisimosti Avenue 4, 220050 Minsk (Belarus); Qiu, C-W [Research Laboratory of Electronics, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139 (United States); Zouhdi, Said [Laboratoire de Genie Electrique de Paris, SUPELEC, Plateau de Moulon 91192, Gif-sur-Yvette (France)], E-mail: eleqc@nus.edu.sg

    2009-11-15

    Based on the concept of the cloak generating function, we propose an implicit transformation-independent method for the required parameters of spherical cloaks without knowing the needed coordinate transformation beforehand. A non-ideal discrete model is used to calculate and optimize the total scattering cross-sections of different profiles of the generating function. A bell-shaped quadratic spherical cloak is found to be the best candidate, which is further optimized by controlling the design parameters involved. Such improved invisibility is steady even when the model is highly discretized.

  11. Method for independent strain and temperature measurement in polymeric tensile test specimen using embedded FBG sensors

    DEFF Research Database (Denmark)

    Pereira, Gilmar Ferreira; McGugan, Malcolm; Mikkelsen, Lars Pilgaard

    2016-01-01

    A novel method to obtain independent strain and temperature measurements using embedded Fibre Bragg Grating (FBG) in polymeric tensile test specimens is presented in this paper. The FBG strain and temperature cross-sensitivity was decoupled using two single mode FBG sensors, which were embedded...... in the specimen material with a certain angle between them. It is demonstrated that, during temperature variation, both FBG sensors show the same signal response. However, for any applied load the signal response is different, which is caused by the different levels of strain acting in each sensor. Equations...... calibration procedure (temperature and strain) was performed to this material-sensor pair, where a calibration error

  12. Computing physical properties with quantum Monte Carlo methods with statistical fluctuations independent of system size.

    Science.gov (United States)

    Assaraf, Roland

    2014-12-01

    We show that the recently proposed correlated sampling without reweighting procedure extends the locality (asymptotic independence of the system size) of a physical property to the statistical fluctuations of its estimator. This makes the approach potentially vastly more efficient for computing space-localized properties in large systems compared with standard correlated methods. A proof is given for a large collection of noninteracting fragments. Calculations on hydrogen chains suggest that this behavior holds not only for systems displaying short-range correlations, but also for systems with long-range correlations.

  13. Comparing results of cultured and uncultured biological methods used in biological phosphorus removal

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Increasing attention has been paid to phosphate-accumulating organisms (PAOs) for their important role in biological phosphorus removal. In this study, microbial communities of PAOs cultivated under different carbon sources (sewage, glucose, and sodium acetate) were investigated and compared through culture-dependent and culture-independent methods, respectively. The results obtained using denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction-amplified 16S rDNA fragments revealed that the diversity of bacteria in a sewage-fed reactor (1#) was much higher than in a glucose-fed one (2#) and a sodium acetate-fed one (3#); there were common PAOs in three reactors fed by different carbon sources. Five strains were separated from three systems by using a phosphate-rich medium; they were from common bacteria isolated and three isolates could not be found in DGGE profile at all. Two isolates had good phosphorus removal ability. When the microbial diversity was studied, the molecular biological method was better than the culture-dependent one. When phosphorus removal characteristics were investigated, culture-dependent approach was more effective. Thus a combination of two methods is necessary to have a comprehensive view of PAOs.

  14. Stable expression of lipocalin-type prostaglandin D synthase in cultured preadipocytes impairs adipogenesis program independently of endogenous prostanoids

    Energy Technology Data Exchange (ETDEWEB)

    Hossain, Mohammad Salim; Chowdhury, Abu Asad; Rahman, Mohammad Sharifur [Department of Life Science and Biotechnology, Shimane University, 1060 Nishikawatsu-cho, Matsue, Shimane 690-8504 (Japan); Nishimura, Kohji [Department of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane University, 1060 Nishikawatsu-cho, Matsue, Shimane 690-8504 (Japan); Jisaka, Mitsuo; Nagaya, Tsutomu [Department of Life Science and Biotechnology, Shimane University, 1060 Nishikawatsu-cho, Matsue, Shimane 690-8504 (Japan); Shono, Fumiaki [Department of Clinical Pharmacy, Faculty of Pharmaceutical Sciences, Tokushima Bunri University, 180 Yamashiro-cho, Tokushima-shi, Tokushima 770-8514 (Japan); Yokota, Kazushige, E-mail: yokotaka@life.shimane-u.ac.jp [Department of Life Science and Biotechnology, Shimane University, 1060 Nishikawatsu-cho, Matsue, Shimane 690-8504 (Japan)

    2012-02-15

    . -- Highlights: Black-Right-Pointing-Pointer Cultured preadipocytes were transfected with sense lipocalin-type PGD synthase. Black-Right-Pointing-Pointer The cloned transfectants had a higher ability to synthesize PGD{sub 2} and PGJ{sub 2} series. Black-Right-Pointing-Pointer The sustained expression of sense L-PGDS hampered the storage of fats. Black-Right-Pointing-Pointer Adipogenesis program was suppressed independently of endogenous prostanoids.

  15. A method for independent modelling in support of regulatory review of dose assessments.

    Science.gov (United States)

    Dverstorp, Björn; Xu, Shulan

    2017-03-22

    Several countries consider geological disposal facilities as the preferred option for spent nuclear fuel due to their potential to provide isolation from the surface environment on very long timescales. In 2011 the Swedish Nuclear Fuel & Waste Management Co. (SKB) submitted a license application for construction of a spent nuclear fuel repository. The disposal method involves disposing spent fuel in copper canisters with a cast iron insert at about 500 m depth in crystalline basement rock, and each canister is surrounded by a buffer of swelling bentonite clay. SKB's license application is supported by a post-closure safety assessment, SR-Site. SR-Site has been reviewed by the Swedish Radiation Safety Authority (SSM) for five years. The main method for review of SKB's license application is document review, which is carried out by SSM's staff and supported by SSM's external experts. The review has proven a challenging task due to its broad scope, complexity and multidisciplinary nature. SSM and its predecessors have, for several decades, been developing independent models to support regulatory reviews of post-closure safety assessments for geological repositories. For the review of SR-Site, SSM has developed a modelling approach with a structured application of independent modelling activities, including replication modelling, use of alternative conceptual models and bounding calculations, to complement the traditional document review. This paper describes this scheme and its application to biosphere and dose assessment modelling. SSM's independent modelling has provided important insights regarding quality and reasonableness of SKB's rather complex biosphere modelling and has helped quantifying conservatisms and highlighting conceptual uncertainty. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Fractured porous medium flow analysis using numerical manifold method with independent covers

    Science.gov (United States)

    Zhang, Qi-Hua; Lin, Shao-Zhong; Xie, Zhi-Qiang; Su, Hai-Dong

    2016-11-01

    Due to the complexity of geometry and the difficulty of mesh discretization of 3D (three-dimensional) blocks cut by complexly distributed fractures, explicitly considering arbitrary fracture network in fractured porous medium (FPM) flow analysis is very challenging for various numerical methods. In this study, we developed a FPM flow model by taking full advantage of numerical manifold method (NMM) with independent covers. With the independent covers, arbitrarily-shaped 3D blocks identified by block-cutting analysis can be directly used as basic computational elements. Along the boundaries of the divided blocks, fractures elements are generated according to the fractures' apertures. Therefore, it is able to handle very complicated fracture network in 3D flow analysis without need to subdivide 3D blocks into computational meshes. In order to refine the meshes, we introduced artificial fractures with same material properties as surrounding rock into a fracture network, without need to coordinate with the shapes of the blocks. We demonstrated our new model on different 2D examples. At last, we applied our model to 2D and 3D examples with complexly distributed fractures, and achieved reasonable results. The results show that our model is very powerful to analyze fluid flow in arbitrarily and complexly fractured rock mass in 3D.

  17. Three-intensity decoy-state method for measurement-device-independent quantum key distribution

    Science.gov (United States)

    Yu, Zong-Wen; Zhou, Yi-Heng; Wang, Xiang-Bin

    2013-12-01

    We study the measurement-device-independent quantum key distribution (MDI-QKD) in practice with limited resources, when there are only three different states in implementing the decoy-state method. We present a tighter explicit formula to estimate the lower bound of the yield of two-single-photon pulses sent by Alice and Bob. Moreover, we show that the bounding of this yield and phase flip error of single-photon pulse pairs can be further improved by using other constraints which can be solved by a simple and explicit program. Our methods here can significantly improve the key rate and the secure distance of MDI-QKD with only three intensities.

  18. 浅谈墨家“修身”思想对廉政文化建设的重要意义%On the Significance of Mohist Thought of Cultivating Morality in Independence Culture

    Institute of Scientific and Technical Information of China (English)

    张遂志

    2014-01-01

    Independence Culture plays an important role in fighting corruption and upholding integrity.In ancient Chinese,Self-discipline culture represented by the thought of Cultivating Morality is fundamental of Independence Culture.Mohist Thought of Cultivating Morality is one root of Independence Culture in ancient Chinese. According to Mohist theory,Cultivating one’s Morality is the root of studying and governing,and claims that self-introspective is basic method,and they insist on that practicing,critical thinking and improving ability are good method to Cultivating Morality.The significance of Mohist thought of cultivating morality in Independence Culture,mainly reflected in adhere to the practice value orientation,self criticism of rationality and setting up the correct friendship culture.%中国古代的廉政文化、修身思想所代表的自律文化对廉政文化有基础作用。墨家“修身”思想是古代廉政文化的重要思想来源之一。墨家将修身作为人们学习和治国理政的根本,提倡“反之身者也”的根本修身方法,坚持实践和批判理性、强调实际能力,提升自我的批判理性和树立正确的交友文化三个方面。

  19. A culture-independent approach to unravel uncultured bacteria and functional genes in a complex microbial community.

    Directory of Open Access Journals (Sweden)

    Yun Wang

    Full Text Available Most microorganisms in nature are uncultured with unknown functionality. Sequence-based metagenomics alone answers 'who/what are there?' but not 'what are they doing and who is doing it and how?'. Function-based metagenomics reveals gene function but is usually limited by the specificity and sensitivity of screening strategies, especially the identification of clones whose functional gene expression has no distinguishable activity or phenotypes. A 'biosensor-based genetic transducer' (BGT technique, which employs a whole-cell biosensor to quantitatively detect expression of inserted genes encoding designated functions, is able to screen for functionality of unknown genes from uncultured microorganisms. In this study, BGT was integrated with Stable isotope probing (SIP-enabled Metagenomics to form a culture-independent SMB toolbox. The utility of this approach was demonstrated in the discovery of a novel functional gene cluster in naphthalene contaminated groundwater. Specifically, metagenomic sequencing of the (13C-DNA fraction obtained by SIP indicated that an uncultured Acidovorax sp. was the dominant key naphthalene degrader in-situ, although three culturable Pseudomonas sp. degraders were also present in the same groundwater. BGT verified the functionality of a new nag2 operon which co-existed with two other nag and two nah operons for naphthalene biodegradation in the same microbial community. Pyrosequencing analysis showed that the nag2 operon was the key functional operon in naphthalene degradation in-situ, and shared homology with both nag operons in Ralstonia sp. U2 and Polaromonas naphthalenivorans CJ2. The SMB toolbox will be useful in providing deep insights into uncultured microorganisms and unravelling their ecological roles in natural environments.

  20. Electron microscopy methods in studies of cultural heritage sites

    Science.gov (United States)

    Vasiliev, A. L.; Kovalchuk, M. V.; Yatsishina, E. B.

    2016-11-01

    The history of the development and application of scanning electron microscopy (SEM), transmission electron microscopy (TEM), and energy-dispersive X-ray microanalysis (EDXMA) in studies of cultural heritage sites is considered. In fact, investigations based on these methods began when electron microscopes became a commercial product. Currently, these methods, being developed and improved, help solve many historical enigmas. To date, electron microscopy combined with microanalysis makes it possible to investigate any object, from parchment and wooden articles to pigments, tools, and objects of art. Studies by these methods have revealed that some articles were made by ancient masters using ancient "nanotechnologies"; hence, their comprehensive analysis calls for the latest achievements in the corresponding instrumental methods and sample preparation techniques.

  1. Considerations for the independent reaction times and step-by-step methods for radiation chemistry simulations

    Science.gov (United States)

    Plante, Ianik; Devroye, Luc

    2017-10-01

    Ionizing radiation interacts with the water molecules of the tissues mostly by ionizations and excitations, which result in the formation of the radiation track structure and the creation of radiolytic species such as H.,.OH, H2, H2O2, and e-aq. After their creation, these species diffuse and may chemically react with the neighboring species and with the molecules of the medium. Therefore radiation chemistry is of great importance in radiation biology. As the chemical species are not distributed homogeneously, the use of conventional models of homogeneous reactions cannot completely describe the reaction kinetics of the particles. Actually, many simulations of radiation chemistry are done using the Independent Reaction Time (IRT) method, which is a very fast technique to calculate radiochemical yields but which do not calculate the positions of the radiolytic species as a function of time. Step-by-step (SBS) methods, which are able to provide such information, have been used only sparsely because these are time-consuming in terms of calculation. Recent improvements in computer performance now allow the regular use of the SBS method in radiation chemistry. The SBS and IRT methods are both based on the Green's functions of the diffusion equation (GFDE). In this paper, several sampling algorithms of the GFDE and for the IRT method are presented. We show that the IRT and SBS methods are exactly equivalent for 2-particles systems for diffusion and partially diffusion-controlled reactions between non-interacting particles. We also show that the results obtained with the SBS simulation method with periodic boundary conditions are in agreement with the predictions by classical reaction kinetics theory, which is an important step towards using this method for modelling of biochemical networks and metabolic pathways involved in oxidative stress. Finally, the first simulation results obtained with the code RITRACKS (Relativistic Ion Tracks) are presented.

  2. Simple and Rapid Method for the Determination of Uric Acid-Independent Antioxidant Capacity

    Directory of Open Access Journals (Sweden)

    Darko Modun

    2011-08-01

    Full Text Available Determination of the relative contribution of uric acid level increases to the total measured antioxidative activity could be very useful for testing antioxidative products and their effect on human health. The aim of this report is to present a simple spectrophotometric method that combines the measurement of total antioxidative capacity of a sample by ferric reducing/antioxidative power (FRAP assay, with the uricase-reaction (specific elimination of uric acid, in order to establish and correct for the contribution of uric acid in FRAP values. We measured FRAP values, with (uric acid-independent antioxidant capacity, TAC-UA and without (total antioxidant capacity, TAC uricase treatment, and expressed it as μmol/L of uric acid equivalents. In such way, it was possible to determine both total and uric acid-independent antioxidant capacity, plasma uric acid (UA, as the difference between TAC and TAC-UA, and the ratio of the uric acid in total antioxidant capacity (UA/TAC.

  3. Quantification of Self Pollution from Two Diesel School Buses using Three Independent Methods

    Science.gov (United States)

    Liu, L.-J. Sally; Phuleria, Harish C.; Webber, Whitney; Davey, Mark; Lawson, Douglas R.; Ireson, Robert G.; Zielinska, Barbara; Ondov, John M.; Weaver, Christopher S.; Lapin, Charles A.; Easter, Michael; Hesterberg, Thomas W.; Larson, Timothy

    2010-01-01

    We monitored two Seattle school buses to quantify the buses’ self pollution using the dual tracers (DT), lead vehicle (LV), and chemical mass balance (CMB) methods. Each bus drove along a residential route simulating stops, with windows closed or open. Particulate matter (PM) and its constituents were monitored in the bus and from a LV. We collected source samples from the tailpipe and crankcase emissions using an on-board dilution tunnel. Concentrations of PM1, ultrafine particle counts, elemental and organic carbon (EC/OC) were higher on the bus than the LV. The DT method estimated that the tailpipe and the crankcase emissions contributed 1.1 and 6.8 μg/m3 of PM2.5 inside the bus, respectively, with significantly higher crankcase self pollution (SP) when windows were closed. Approximately two-thirds of in-cabin PM2.5 originated from background sources. Using the LV approach, SP estimates from the EC and the active personal DataRAM (pDR) measurements correlated well with the DT estimates for tailpipe and crankcase emissions, respectively, although both measurements need further calibration for accurate quantification. CMB results overestimated SP from the DT method but confirmed crankcase emissions as the major SP source. We confirmed buses’ SP using three independent methods and quantified crankcase emissions as the dominant contributor. PMID:20694046

  4. The IRIDICA BAC BSI Assay: Rapid, Sensitive and Culture-Independent Identification of Bacteria and Candida in Blood.

    Directory of Open Access Journals (Sweden)

    David Metzgar

    Full Text Available Bloodstream infection (BSI and sepsis are rising in incidence throughout the developed world. The spread of multi-drug resistant organisms presents increasing challenges to treatment. Surviving BSI is dependent on rapid and accurate identification of causal organisms, and timely application of appropriate antibiotics. Current culture-based methods used to detect and identify agents of BSI are often too slow to impact early therapy and may fail to detect relevant organisms in many positive cases. Existing methods for direct molecular detection of microbial DNA in blood are limited in either sensitivity (likely the result of small sample volumes or in breadth of coverage, often because the PCR primers and probes used target only a few specific pathogens. There is a clear unmet need for a sensitive molecular assay capable of identifying the diverse bacteria and yeast associated with BSI directly from uncultured whole blood samples. We have developed a method of extracting DNA from larger volumes of whole blood (5 ml per sample, amplifying multiple widely conserved bacterial and fungal genes using a mismatch- and background-tolerant PCR chemistry, and identifying hundreds of diverse organisms from the amplified fragments on the basis of species-specific genetic signatures using electrospray ionization mass spectrometry (PCR/ESI-MS. We describe the analytical characteristics of the IRIDICA BAC BSI Assay and compare its pre-clinical performance to current standard-of-care methods in a collection of prospectively collected blood specimens from patients with symptoms of sepsis. The assay generated matching results in 80% of culture-positive cases (86% when common contaminants were excluded from the analysis, and twice the total number of positive detections. The described method is capable of providing organism identifications directly from uncultured blood in less than 8 hours.The IRIDICA BAC BSI Assay is not available in the United States.

  5. A safety culture assessment by mixed methods at a public maternity and infant hospital in China

    Directory of Open Access Journals (Sweden)

    Listyowardojo TA

    2017-07-01

    Full Text Available Tita Alissa Listyowardojo,1 Xiaoling Yan,2,3 Stephen Leyshon,1 Bobbie Ray-Sannerud,1 Xin Yan Yu,4 Kai Zheng,4 Tao Duan2,3 1Life Sciences Program, Group Technology and Research, DNV GL, Hovik, Norway; 2Quality and Safety Department, Shanghai First Maternity and Infant Hospital, 3Tongji University School of Medicine, Shanghai, 4Healthcare Department, Business Assurance, DNV GL, Beijing, China Objective: To assess safety culture at a public maternity hospital in Shanghai, China, using a sequential mixed methods approach. The study was part of a bigger study looking at the application of the mixed methods approach to assess safety culture in health care in different organizations and countries.Methodology: A mixed methods approach was utilized by first distributing the Safety Attitudes Questionnaire measuring six safety culture dimensions and five independent items to all hospital staff (n=1482 working in 18 departments at a single hospital. Afterward, semistructured interviews were conducted using convenience sampling, where 48 hospital staff from nine departments at the same hospital were individually interviewed.Results: The survey received a response rate of 96%. The survey findings show significant differences between the hospital departments in almost all safety culture dimensions and independent items. Similarly, the interview findings revealed that there were different, competing priorities between departments perceived to result in a reduced quality of collaboration and bottlenecks in care delivery. Another major finding was that staff who worked more hours per week would perceive working conditions significantly more negatively. Issues related to working conditions were also the most common concerns discussed in the interviews, especially the issue on high workload. High workload was also reflected in the fact that 91.45% of survey respondents reported that they worked 40 hours or longer per week. Finally, interview findings complemented

  6. Introducing an Efficient Method for Scheduling Independent Tasks in Grid Environment using Meta-Heuristic Algorithms

    Directory of Open Access Journals (Sweden)

    Masoud Shirzadi

    2015-11-01

    Full Text Available Since the dynamicity and inhomogeneity of resources complicates scheduling, it is not possible to use accurate scheduling algorithms. Therefore, many studies focus on heuristic algorithms like the artificial bee colony algorithm. Since, the artificial bee colony algorithm searches the problem space locally and has a poor performance in global search; global search algorithms like genetic algorithms should also be used to overcome this drawback. This study proposes a scheduling algorithm, which is combination of the genetic and artificial bee colony algorithms for the independent scheduling problem in a computing grid. This study aims to reduce the maximum total scheduling time. Simulation results indicate that the proposed algorithm reduces the maximum execution time (makespan by 10% in comparison to the compared methods.

  7. Testing a scale-independent method to measure the mass of black holes

    CERN Document Server

    Gliozzi, M; Satyapal, S; Price, D M; Jang, I

    2011-01-01

    Estimating the black hole mass at the center of galaxies is a fundamental step not only for understanding the physics of accretion, but also for the cosmological evolution of galaxies. Recently a new method, based solely on X-ray data, was successfully applied to determine the black hole mass in Galactic systems. Since X-rays are thought to be produced via Comptonization process both in stellar and supermassive black holes, in principle, the same method may be applied to estimate the mass in supermassive black holes. In this work we test this hypothesis by performing a systematic analysis of a sample of AGNs, whose black hole mass has been already determined via reverberation mapping and which possess high quality XMM-Newton archival data. The good agreement obtained between the black hole masses derived with this novel scaling technique and the reverberation mapping values suggests that this method is robust and works equally well on stellar and supermassive black holes, making it a truly scale-independent t...

  8. The hydrogen tunneling splitting in malonaldehyde: A full-dimensional time-independent quantum mechanical method

    Science.gov (United States)

    Wu, Feng; Ren, Yinghui; Bian, Wensheng

    2016-08-01

    The accurate time-independent quantum dynamics calculations on the ground-state tunneling splitting of malonaldehyde in full dimensionality are reported for the first time. This is achieved with an efficient method developed by us. In our method, the basis functions are customized for the hydrogen transfer process which has the effect of greatly reducing the size of the final Hamiltonian matrix, and the Lanczos method and parallel strategy are used to further overcome the memory and central processing unit time bottlenecks. The obtained ground-state tunneling splitting of 24.5 cm-1 is in excellent agreement with the benchmark value of 23.8 cm-1 computed with the full-dimensional, multi-configurational time-dependent Hartree approach on the same potential energy surface, and we estimate that our reported value has an uncertainty of less than 0.5 cm-1. Moreover, the role of various vibrational modes strongly coupled to the hydrogen transfer process is revealed.

  9. Prognostic breast cancer signature identified from 3D culture model accurately predicts clinical outcome across independent datasets

    Energy Technology Data Exchange (ETDEWEB)

    Martin, Katherine J.; Patrick, Denis R.; Bissell, Mina J.; Fournier, Marcia V.

    2008-10-20

    One of the major tenets in breast cancer research is that early detection is vital for patient survival by increasing treatment options. To that end, we have previously used a novel unsupervised approach to identify a set of genes whose expression predicts prognosis of breast cancer patients. The predictive genes were selected in a well-defined three dimensional (3D) cell culture model of non-malignant human mammary epithelial cell morphogenesis as down-regulated during breast epithelial cell acinar formation and cell cycle arrest. Here we examine the ability of this gene signature (3D-signature) to predict prognosis in three independent breast cancer microarray datasets having 295, 286, and 118 samples, respectively. Our results show that the 3D-signature accurately predicts prognosis in three unrelated patient datasets. At 10 years, the probability of positive outcome was 52, 51, and 47 percent in the group with a poor-prognosis signature and 91, 75, and 71 percent in the group with a good-prognosis signature for the three datasets, respectively (Kaplan-Meier survival analysis, p<0.05). Hazard ratios for poor outcome were 5.5 (95% CI 3.0 to 12.2, p<0.0001), 2.4 (95% CI 1.6 to 3.6, p<0.0001) and 1.9 (95% CI 1.1 to 3.2, p = 0.016) and remained significant for the two larger datasets when corrected for estrogen receptor (ER) status. Hence the 3D-signature accurately predicts breast cancer outcome in both ER-positive and ER-negative tumors, though individual genes differed in their prognostic ability in the two subtypes. Genes that were prognostic in ER+ patients are AURKA, CEP55, RRM2, EPHA2, FGFBP1, and VRK1, while genes prognostic in ER patients include ACTB, FOXM1 and SERPINE2 (Kaplan-Meier p<0.05). Multivariable Cox regression analysis in the largest dataset showed that the 3D-signature was a strong independent factor in predicting breast cancer outcome. The 3D-signature accurately predicts breast cancer outcome across multiple datasets and holds prognostic

  10. Analysis of novel geometry-independent method for dialysis access pressure-flow monitoring

    Directory of Open Access Journals (Sweden)

    Panduranga Harsha

    2008-11-01

    Full Text Available Abstract Background End-stage renal disease (ESRD confers a large health-care burden for the United States, and the morbidity associated with vascular access failure has stimulated research into detection of vascular access stenosis and low flow prior to thrombosis. We present data investigating the possibility of using differential pressure (ΔP monitoring to estimate access flow (Q for dialysis access monitoring, with the goal of utilizing micro-electro-mechanical systems (MEMS pressure sensors integrated within the shaft of dialysis needles. Methods A model of the arteriovenous graft fluid circuit was used to study the relationship between Q and the ΔP between two dialysis needles placed 2.5–20.0 cm apart. Tubing was varied to simulate grafts with inner diameters of 4.76–7.95 mm. Data were compared with values from two steady-flow models. These results, and those from computational fluid dynamics (CFD modeling of ΔP as a function of needle position, were used to devise and test a method of estimating Q using ΔP and variable dialysis pump speeds (variable flow that diminishes dependence on geometric factors and fluid characteristics. Results In the fluid circuit model, ΔP increased with increasing volume flow rate and with increasing needle-separation distance. A nonlinear model closely predicts this ΔP-Q relationship (R2 > 0.98 for all graft diameters and needle-separation distances tested. CFD modeling suggested turbulent needle effects are greatest within 1 cm of the needle tip. Utilizing linear, quadratic and combined variable flow algorithms, dialysis access flow was estimated using geometry-independent models and an experimental dialysis system with the pressure sensors separated from the dialysis needle tip by distances ranging from 1 to 5 cm. Real-time ΔP waveform data were also observed during the mock dialysis treatment, which may be useful in detecting low or reversed flow within the access. Conclusion With further

  11. LEA Detection and Tracking Method for Color-Independent Visual-MIMO.

    Science.gov (United States)

    Kim, Jai-Eun; Kim, Ji-Won; Kim, Ki-Doo

    2016-07-02

    Communication performance in the color-independent visual-multiple input multiple output (visual-MIMO) technique is deteriorated by light emitting array (LEA) detection and tracking errors in the received image because the image sensor included in the camera must be used as the receiver in the visual-MIMO system. In this paper, in order to improve detection reliability, we first set up the color-space-based region of interest (ROI) in which an LEA is likely to be placed, and then use the Harris corner detection method. Next, we use Kalman filtering for robust tracking by predicting the most probable location of the LEA when the relative position between the camera and the LEA varies. In the last step of our proposed method, the perspective projection is used to correct the distorted image, which can improve the symbol decision accuracy. Finally, through numerical simulation, we show the possibility of robust detection and tracking of the LEA, which results in a symbol error rate (SER) performance improvement.

  12. Maximum ikelihood estimation for the double-count method with independent observers

    Science.gov (United States)

    Manly, Bryan F.J.; McDonald, Lyman L.; Garner, Gerald W.

    1996-01-01

    Data collected under a double-count protocol during line transect surveys were analyzed using new maximum likelihood methods combined with Akaike's information criterion to provide estimates of the abundance of polar bear (Ursus maritimus Phipps) in a pilot study off the coast of Alaska. Visibility biases were corrected by modeling the detection probabilities using logistic regression functions. Independent variables that influenced the detection probabilities included perpendicular distance of bear groups from the flight line and the number of individuals in the groups. A series of models were considered which vary from (1) the simplest, where the probability of detection was the same for both observers and was not affected by either distance from the flight line or group size, to (2) models where probability of detection is different for the two observers and depends on both distance from the transect and group size. Estimation procedures are developed for the case when additional variables may affect detection probabilities. The methods are illustrated using data from the pilot polar bear survey and some recommendations are given for design of a survey over the larger Chukchi Sea between Russia and the United States.

  13. Microseismic imaging using a source-independent full-waveform inversion method

    KAUST Repository

    Wang, Hanchen

    2016-09-06

    Using full waveform inversion (FWI) to locate microseismic and image microseismic events allows for an automatic process (free of picking) that utilizes the full wavefield. However, waveform inversion of microseismic events faces incredible nonlinearity due to the unknown source location (space) and function (time). We develop a source independent FWI of microseismic events to invert for the source image, source function and the velocity model. It is based on convolving reference traces with the observed and modeled data to mitigate the effect of an unknown source ignition time. The adjoint-state method is used to derive the gradient for the source image, source function and velocity updates. The extended image for source wavelet in z axis is extracted to check the accuracy of the inverted source image and velocity model. Also the angle gather is calculated to see if the velocity model is correct. By inverting for all the source image, source wavelet and the velocity model, the proposed method produces good estimates of the source location, ignition time and the background velocity for part of the SEG overthrust model.

  14. Type 2 Diabetes Biomarkers of Human Gut Microbiota Selected via Iterative Sure Independent Screening Method.

    Directory of Open Access Journals (Sweden)

    Lihua Cai

    Full Text Available Type 2 diabetes, which is a complex metabolic disease influenced by genetic and environment, has become a worldwide problem. Previous published results focused on genetic components through genome-wide association studies that just interpret this disease to some extent. Recently, two research groups published metagenome-wide association studies (MGWAS result that found meta-biomarkers related with type 2 diabetes. However, One key problem of analyzing genomic data is that how to deal with the ultra-high dimensionality of features. From a statistical viewpoint it is challenging to filter true factors in high dimensional data. Various methods and techniques have been proposed on this issue, which can only achieve limited prediction performance and poor interpretability. New statistical procedure with higher performance and clear interpretability is appealing in analyzing high dimensional data. To address this problem, we apply an excellent statistical variable selection procedure called iterative sure independence screening to gene profiles that obtained from metagenome sequencing, and 48/24 meta-markers were selected in Chinese/European cohorts as predictors with 0.97/0.99 accuracy in AUC (area under the curve, which showed a better performance than other model selection methods, respectively. These results demonstrate the power and utility of data mining technologies within the large-scale and ultra-high dimensional genomic-related dataset for diagnostic and predictive markers identifying.

  15. The East-West Method: An Exposure-independent Method to Search for Large-scale Anisotropies of Cosmic Rays

    Science.gov (United States)

    Bonino, R.; Alekseenko, V. V.; Deligny, O.; Ghia, P. L.; Grigat, M.; Letessier-Selvon, A.; Lyberis, H.; Mollerach, S.; Over, S.; Roulet, E.

    2011-09-01

    The measurement of large-scale anisotropies in cosmic ray arrival directions at energies above 1013 eV is performed through the detection of extensive air showers (EAS) produced by cosmic ray interactions in the atmosphere. The observed anisotropies are small, so accurate measurements require small statistical uncertainties, i.e., large data sets. These can be obtained by employing ground detector arrays with large extensions (from 104 to 109 m2) and long operation time (up to 20 years). The control of such arrays is challenging and spurious variations in the counting rate due to instrumental effects (e.g., data taking interruptions or changes in the acceptance) and atmospheric effects (e.g., air temperature and pressure effects on EAS development) are usually present. These modulations must be corrected very precisely before performing standard anisotropy analyses, i.e., harmonic analysis of the counting rate versus local sidereal time. In this paper we discuss an alternative method to measure large-scale anisotropies, the "East-West method." It was originally proposed in the 1940s to study asymmetries in the flux of solar cosmic rays and later applied by Nagashima et al. to EAS at higher energies. It is a differential method, as it is based on the analysis of the difference of the counting rates in the east and west directions. Besides explaining the principle, we present here its mathematical derivation, showing that the method is largely independent of experimental effects, that is, it does not require corrections for acceptance and/or for atmospheric effects. We explain the use of the method to derive the amplitude and phase of the anisotropy and demonstrate its power under different conditions of detector operation.

  16. Microbial diversity and dynamics throughout manufacturing and ripening of surface ripened semi-hard Danish Danbo cheeses investigated by culture-independent techniques

    DEFF Research Database (Denmark)

    Ryssel, Mia; Johansen, Pernille; Abu Al-Soud, Waleed

    2015-01-01

    Microbial successions on the surface and in the interior of surface ripened semi-hard Danish Danbo cheeses were investigated by culture-dependent and -independent techniques. Culture-independent detection of microorganisms was obtained by denaturing gradient gel electrophoresis (DGGE) and pyroseq.......9%±7.8% of the OTUs throughout the cheese processing. The microbial dynamics described at genus level in this study add to a comprehensive understanding of the complex microbiota existing especially on surface ripened semi-hard cheeses.......Microbial successions on the surface and in the interior of surface ripened semi-hard Danish Danbo cheeses were investigated by culture-dependent and -independent techniques. Culture-independent detection of microorganisms was obtained by denaturing gradient gel electrophoresis (DGGE....... As expected, microbial profiles of the surface and the interior of the cheeses diverged. During cheese production pyrosequencing determined Lactococcus as the dominating genus on cheese surfaces, representing on average 94.7%±2.1% of the OTUs. At day 6 Lactococcus spp. declined to 10.0% of the OTUs, whereas...

  17. Developing a Culture of Enquiry-Based, Independent Learning in a Research-Led Institution: Findings from a Survey of Pedagogic Practice

    Science.gov (United States)

    McLinden, Mike; Edwards, Corony

    2011-01-01

    This paper reports select findings from an institutional survey designed to support long-term strategic developments at a research-led institution in the UK. These developments include a revised Learning and Teaching Strategy that has at its core the promotion of a "cross-institutional culture of enquiry-based, independent learning". The…

  18. Novel and Unexpected Microbial Diversity in Acid Mine Drainage in Svalbard (78° N, Revealed by Culture-Independent Approaches

    Directory of Open Access Journals (Sweden)

    Antonio García-Moyano

    2015-10-01

    Full Text Available Svalbard, situated in the high Arctic, is an important past and present coal mining area. Dozens of abandoned waste rock piles can be found in the proximity of Longyearbyen. This environment offers a unique opportunity for studying the biological control over the weathering of sulphide rocks at low temperatures. Although the extension and impact of acid mine drainage (AMD in this area is known, the native microbial communities involved in this process are still scarcely studied and uncharacterized. Several abandoned mining areas were explored in the search for active AMD and a culture-independent approach was applied with samples from two different runoffs for the identification and quantification of the native microbial communities. The results obtained revealed two distinct microbial communities. One of the runoffs was more extreme with regards to pH and higher concentration of soluble iron and heavy metals. These conditions favored the development of algal-dominated microbial mats. Typical AMD microorganisms related to known iron-oxidizing bacteria (Acidithiobacillus ferrivorans, Acidobacteria and Actinobacteria dominated the bacterial community although some unexpected populations related to Chloroflexi were also significant. No microbial mats were found in the second area. The geochemistry here showed less extreme drainage, most likely in direct contact with the ore under the waste pile. Large deposits of secondary minerals were found and the presence of iron stalks was revealed by microscopy analysis. Although typical AMD microorganisms were also detected here, the microbial community was dominated by other populations, some of them new to this type of system (Saccharibacteria, Gallionellaceae. These were absent or lowered in numbers the farther from the spring source and they could represent native populations involved in the oxidation of sulphide rocks within the waste rock pile. This environment appears thus as a highly interesting

  19. Novel and Unexpected Microbial Diversity in Acid Mine Drainage in Svalbard (78° N), Revealed by Culture-Independent Approaches.

    Science.gov (United States)

    García-Moyano, Antonio; Austnes, Andreas Erling; Lanzén, Anders; González-Toril, Elena; Aguilera, Ángeles; Øvreås, Lise

    2015-10-13

    Svalbard, situated in the high Arctic, is an important past and present coal mining area. Dozens of abandoned waste rock piles can be found in the proximity of Longyearbyen. This environment offers a unique opportunity for studying the biological control over the weathering of sulphide rocks at low temperatures. Although the extension and impact of acid mine drainage (AMD) in this area is known, the native microbial communities involved in this process are still scarcely studied and uncharacterized. Several abandoned mining areas were explored in the search for active AMD and a culture-independent approach was applied with samples from two different runoffs for the identification and quantification of the native microbial communities. The results obtained revealed two distinct microbial communities. One of the runoffs was more extreme with regards to pH and higher concentration of soluble iron and heavy metals. These conditions favored the development of algal-dominated microbial mats. Typical AMD microorganisms related to known iron-oxidizing bacteria (Acidithiobacillus ferrivorans, Acidobacteria and Actinobacteria) dominated the bacterial community although some unexpected populations related to Chloroflexi were also significant. No microbial mats were found in the second area. The geochemistry here showed less extreme drainage, most likely in direct contact with the ore under the waste pile. Large deposits of secondary minerals were found and the presence of iron stalks was revealed by microscopy analysis. Although typical AMD microorganisms were also detected here, the microbial community was dominated by other populations, some of them new to this type of system (Saccharibacteria, Gallionellaceae). These were absent or lowered in numbers the farther from the spring source and they could represent native populations involved in the oxidation of sulphide rocks within the waste rock pile. This environment appears thus as a highly interesting field of potential

  20. Inhibition of bacterial growth in sweet cheese whey by carbon dioxide as determined by culture-independent community profiling.

    Science.gov (United States)

    Lo, Raquel; Xue, Tian; Weeks, Mike; Turner, Mark S; Bansal, Nidhi

    2016-01-18

    Whey is a valuable co-product from cheese making that serves as a raw material for a wide range of products. Its rich nutritional content lends itself to rapid spoilage, thus it typically needs to be pasteurised and refrigerated promptly. Despite the extensive literature on milk spoilage bacteria, little is known about the spoilage bacteria of whey. The utility of carbon dioxide (CO2) to extend the shelf-life of raw milk and cottage cheese has been well established, but its application in whey preservation has not yet been explored. This study aims to characterise the microbial populations of fresh and spoiled sweet whey by culture-independent community profiling using 454 pyrosequencing of 16S rRNA gene amplicons and to determine whether carbonation is effective in inhibiting bacterial growth in sweet whey. The microbiota of raw Cheddar and Mozzarella whey was dominated by cheese starter bacteria. After pasteurisation, two out of the three samples studied became dominated by diverse environmental bacteria from various phyla, with Proteobacteria being the most dominant. Diverse microbial profiles were maintained until spoilage occurred, when the entire population was dominated by just one or two genera. Whey spoilage bacteria were found to be similar to those of milk. Pasteurised Cheddar and Mozzarella whey was spoiled by Bacillus sp. or Pseudomonas sp., and raw Mozzarella whey was spoiled by Pseudomonas sp., Serratia sp., and other members of the Enterobacteriaceae family. CO2 was effective in inhibiting bacterial growth of pasteurised Cheddar and Mozzarella whey stored at 15°C and raw Mozzarella whey stored at 4°C. The spoilage bacteria of the carbonated samples were similar to those of the non-carbonated controls.

  1. Culture-independent characterization of bacteria and fungi in a poultry bioaerosol using pyrosequencing: a new approach.

    Science.gov (United States)

    Nonnenmann, M W; Bextine, B; Dowd, S E; Gilmore, K; Levin, J L

    2010-12-01

    Work in animal production facilities often results in exposure to organic dusts. Previous studies have documented decreases in pulmonary function and lung inflammation among workers exposed to organic dust in the poultry industry. Bacteria and fungi have been reported as components of the organic dust produced in poultry facilities. To date, little is known about the diversity and concentration of bacteria and fungi inside poultry buildings. All previous investigations have utilized culture-based methods for analysis that identify only biota cultured on selected media. The bacterial tag-encoded flexible (FLX) amplicon pyrosequencing (bTEFAP) and fungal tag-encoded flexible (FLX) amplicon pyrosequencing (fTEFAP) are modern and comprehensive approaches for determining biodiversity of microorganisms and have not previously been used to provide characterization of exposure to microorganisms in an occupational environment. This article illustrates the potential application of this novel technique in occupational exposure assessment as well as other settings. An 8-hr area sample was collected using an Institute of Medicine inhalable sampler attached to a mannequin in a poultry confinement building. The sample was analyzed using bTEFAP and fTEFAP. Of the bacteria and fungi detected, 116 and 39 genera were identified, respectively. Among bacteria, Staphylococcus cohnii was present in the highest proportion (23%). The total inhalable bacteria concentration was estimated to be 7503 cells/m³. Among the fungi identified, Sagenomella sclerotialis was present in the highest proportion (37%). Aspergillus ochraceus and Penicillium janthinellum were also present in high proportions. The total inhalable fungi concentration was estimated to be 1810 cells/m³. These estimates are lower than what has been reported by others using standard epifluorescence microscope methods. However, no study has used non-culture-based techniques, such as bTEFAP and fTEFAP, to evaluate bacteria and

  2. A platform-independent method to reduce CT truncation artifacts using discriminative dictionary representations.

    Science.gov (United States)

    Chen, Yang; Budde, Adam; Li, Ke; Li, Yinsheng; Hsieh, Jiang; Chen, Guang-Hong

    2017-01-01

    When the scan field of view (SFOV) of a CT system is not large enough to enclose the entire cross-section of the patient, or the patient needs to be positioned partially outside the SFOV for certain clinical applications, truncation artifacts often appear in the reconstructed CT images. Many truncation artifact correction methods perform extrapolations of the truncated projection data based on certain a priori assumptions. The purpose of this work was to develop a novel CT truncation artifact reduction method that directly operates on DICOM images. The blooming of pixel values associated with truncation was modeled using exponential decay functions, and based on this model, a discriminative dictionary was constructed to represent truncation artifacts and nonartifact image information in a mutually exclusive way. The discriminative dictionary consists of a truncation artifact subdictionary and a nonartifact subdictionary. The truncation artifact subdictionary contains 1000 atoms with different decay parameters, while the nonartifact subdictionary contains 1000 independent realizations of Gaussian white noise that are exclusive with the artifact features. By sparsely representing an artifact-contaminated CT image with this discriminative dictionary, the image was separated into a truncation artifact-dominated image and a complementary image with reduced truncation artifacts. The artifact-dominated image was then subtracted from the original image with an appropriate weighting coefficient to generate the final image with reduced artifacts. This proposed method was validated via physical phantom studies and retrospective human subject studies. Quantitative image evaluation metrics including the relative root-mean-square error (rRMSE) and the universal image quality index (UQI) were used to quantify the performance of the algorithm. For both phantom and human subject studies, truncation artifacts at the peripheral region of the SFOV were effectively reduced, revealing

  3. Bio-Augmentation of Cupriavidus sp. CY-1 into 2,4-D Contaminated Soil: Microbial Community Analysis by Culture Dependent and Independent Techniques.

    Science.gov (United States)

    Chang, Young-Cheol; Reddy, M Venkateswar; Umemoto, Honoka; Sato, Yuki; Kang, Mi-Hye; Yajima, Yuka; Kikuchi, Shintaro

    2015-01-01

    In the present study, a 2,4-dichlorophenoxyacetic acid (2,4-D) degrading bacterial strain CY-1 was isolated from the forest soil. Based on physiological, biochemical and 16S rRNA gene sequence analysis it was identified as Cupriavidus sp. CY-1. Further 2,4-D degradation experiments at different concentrations (200 to 800 mg l(-1)) were carried out using CY-1. Effect of NaCl and KNO3 on 2,4-D degradation was also evaluated. Degradation of 2,4-D and the metabolites produced during degradation process were analyzed using high pressure liquid chromatography (HPLC) and GC-MS respectively. The amount of chloride ions produced during the 2,4-D degradation were analyzed by Ion chromatography (IC) and it is stoichiometric with 2,4-D dechlorination. Furthermore two different types of soils collected from two different sources were used for 2,4-D degradation studies. The isolated strain CY-1 was bio-augmented into 2,4-D contaminated soils to analyze its degradation ability. Culture independent methods like denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP), and culture dependent methods like colony forming units (CFU) and most probable number (MPN) were used to analyze the survivability of strain CY-1 in contaminated soil. Results of T-RFLP were coincident with the DGGE analysis. From the DGGE, T-RFLP, MPN and HPLC results it was concluded that strain CY-1 effectively degraded 2,4-D without disturbing the ecosystem of soil indigenous microorganisms.

  4. Bio-Augmentation of Cupriavidus sp. CY-1 into 2,4-D Contaminated Soil: Microbial Community Analysis by Culture Dependent and Independent Techniques.

    Directory of Open Access Journals (Sweden)

    Young-Cheol Chang

    Full Text Available In the present study, a 2,4-dichlorophenoxyacetic acid (2,4-D degrading bacterial strain CY-1 was isolated from the forest soil. Based on physiological, biochemical and 16S rRNA gene sequence analysis it was identified as Cupriavidus sp. CY-1. Further 2,4-D degradation experiments at different concentrations (200 to 800 mg l(-1 were carried out using CY-1. Effect of NaCl and KNO3 on 2,4-D degradation was also evaluated. Degradation of 2,4-D and the metabolites produced during degradation process were analyzed using high pressure liquid chromatography (HPLC and GC-MS respectively. The amount of chloride ions produced during the 2,4-D degradation were analyzed by Ion chromatography (IC and it is stoichiometric with 2,4-D dechlorination. Furthermore two different types of soils collected from two different sources were used for 2,4-D degradation studies. The isolated strain CY-1 was bio-augmented into 2,4-D contaminated soils to analyze its degradation ability. Culture independent methods like denaturing gradient gel electrophoresis (DGGE and terminal restriction fragment length polymorphism (T-RFLP, and culture dependent methods like colony forming units (CFU and most probable number (MPN were used to analyze the survivability of strain CY-1 in contaminated soil. Results of T-RFLP were coincident with the DGGE analysis. From the DGGE, T-RFLP, MPN and HPLC results it was concluded that strain CY-1 effectively degraded 2,4-D without disturbing the ecosystem of soil indigenous microorganisms.

  5. Characterization of microbes in prosthetic joint specimens by culture-independent molecular methods

    DEFF Research Database (Denmark)

    Thomsen, Trine Rolighed; Xu, Yijuan; Rudkjøbing, Vibeke Børsholt

    (qPCR). In order to reveal possible heterogeneous distribution of the microbes, multiple biopsies were taken from wounds, biofilm was scraped from internal and external catheter surfaces, and during revision arthroplasty several specimen types (joint fluid, tissue biopsy, bone biopsy and prosthesis...

  6. Microbial ecology of artisanal italian cheese: Molecular microbial characterization by culture-independent method

    Energy Technology Data Exchange (ETDEWEB)

    Colombo, E.; Scarpellini, M.; Franzatti, L.; Dioguardi, L.

    2009-07-01

    Present study will treat the next topics: ecology of the natural and man made environments and functional diversity of bacteria. The microbial communities in artisanal goat cheeses produced in mountain pastures (typical farms) in Piemonte mountain (North of Italy) change a lot during precessing and ripening time. Moreover cheese microbial ecosystems are different in each small dairy because adventitious microflora can come from the environment and contamination the milk before the cheese making process and the product during manufacture and ripening. (Author)

  7. Determination of biodegradation potential by two culture-independent methods in PAH-contaminated soils.

    Science.gov (United States)

    Moon, H S; Kahng, H-Y; Kim, J Y; Kukor, J J; Nam, K

    2006-04-01

    Biodegradation potentials of polycyclic aromatic hydrocarbons (PAHs) were determined with soil samples collected from various depths of a PAH-contaminated site and of a site nearby where PAHs were not found. Putative dioxygenase genes were amplified by a primer set specific for initial dioxygenases and identified by web-based database homology search. They were further categorized into several groups of which four dioxygenases were selected as probes for DNA hybridization. The hybridization signals according to the presence of putative dioxygenases were positively related to the extent of PAH contamination. However, the signal intensities varied depending on the probes hybridized and moreover were not consistent with PAH biodegradation activities determined by CO2 evolution. Despite widely accepted advantages of molecular biodegradation assessment, our data clearly present the variations of assessment results depending on the genetic information used and suggest that the methodology may tend to underestimate the real biodegradation capacity of a site probably due to the limited dioxygenase database available at the moment. Therefore, the molecular assessment of biodegradation potential should involve a very careful primer and probe design and an extensive microbiological examination of a site of interest to accurately delineate the biodegradation potential of the site.

  8. The Teaching Methods of Cultural Factors in The Classroom

    Institute of Scientific and Technical Information of China (English)

    Jia Mengyang

    2014-01-01

    Culture knowledge plays an important role in linguistic proficiency and currently most teaching activities are stil happened inthe traditionalclassroom. So this paper introducedsome ofthe practicalteachingmethods ofChinese culture inthe Chinese language classroom.

  9. Methods for reducing the ammonia in hybridoma cell cultures.

    Science.gov (United States)

    Capiaumont, J; Legrand, C; Carbonell, D; Dousset, B; Belleville, F; Nabet, P

    1995-02-21

    The factors which limit the proliferation of eukaryotic cells in vitro are still not well known. Ammonia is believed to be toxic for mammalian cell proliferation and secretion. We have tried two approaches to reducing the ammonia in the medium. We first limited the ammonia produced by the cells by replacing glutamine by glutamate. Then, we used two chemical engineering methods to eliminate accumulated ammonia. In one the used medium was passed through a natural cation exchanger: the clinoptilolite. In the other, the culture medium was passed through a hydrophobic microporous hollow fiber module. Replacing the glutamine by glutamate reduced the medium ammonia concentration. The physicochemical removal of ammonia induced a better cell growth, but not a better specific antibody secretion.

  10. Methods for the Organogenesis of Skeletal Muscle in Tissue Culture

    Science.gov (United States)

    Vandenburgh, Herman; Shansky, Janet; DelTatto, Michael; Chromiak, Joseph

    1997-01-01

    Skeletal muscle structure is regulated by many factors, including nutrition, hormones, electrical activity, and tension. The muscle cells are subjected to both passive and active mechanical forces at all stages of development and these forces play important but poorly understood roles in regulating muscle organogenesis and growth. For example, during embryogenesis, the rapidly growing skeleton places large passive mechanical forces on the attached muscle tissue. These forces not only help to organize the proliferating mononucleated myoblasts into the oriented, multinucleated myofibers of a functional muscle but also tightly couple the growth rate of muscle to that of bone. Postnatally, the actively contracting, innervated muscle fibers are subjected to different patterns of active and passive tensions which regulate longitudinal and cross sectional myofiber growth. These mechanically-induced organogenic processes have been difficult to study under normal tissue culture conditions, resulting in the development of numerous methods and specialized equipment to simulate the in vivo mechanical environment.These techniques have led to the "engineering" of bioartificial muscles (organoids) which display many of the characteristics of in vivo muscle including parallel arrays of postmitotic fibers organized into fascicle-like structures with tendon-like ends. They are contractile, express adult isoforms of contractile proteins, perform directed work, and can be maintained in culture for long periods. The in vivo-like characteristics and durability of these muscle organoids make them useful for long term in vitro studies on mechanotransduction mechanisms and on muscle atrophy induced by decreased tension. In this report, we described a simple method for generating muscle organoids from either primary embrionic avain or neonatal rodent myoblasts.

  11. A Capacity-Restraint Transit Assignment Model When a Predetermination Method Indicates the Invalidity of Time Independence

    Directory of Open Access Journals (Sweden)

    Haoyang Ding

    2015-01-01

    Full Text Available The statistical independence of time of every two adjacent bus links plays a crucial role in deciding the feasibility of using many mathematical models to analyze urban transit networks. Traditional research generally ignores the time independence that acts as the ground of their models. Assumption is usually made that time independence of every two adjacent links is sound. This is, however, actually groundless and probably causes problematic conclusions reached by corresponding models. Many transit assignment models such as multinomial probit-based models lose their effects when the time independence is not valid. In this paper, a simple method to predetermine the time independence is proposed. Based on the predetermination method, a modified capacity-restraint transit assignment method aimed at engineering practice is put forward and tested through a small contrived network and a case study in Nanjing city, China, respectively. It is found that the slope of regression equation between the mean and standard deviation of normal distribution acts as the indicator of time independence at the same time. Besides, our modified assignment method performs better than the traditional one with more reasonable results while keeping the property of simplicity well.

  12. Culture-independent sequence analysis of Chlamydia trachomatis in urogenital specimens identifies regions of recombination and in-patient sequence mutations.

    Science.gov (United States)

    Putman, Timothy E; Suchland, Robert J; Ivanovitch, John D; Rockey, Daniel D

    2013-10-01

    A culture-independent genome sequencing approach was developed and used to examine genomic variability in Chlamydia trachomatis-positive specimens that were collected from patients in the Seattle, WA, USA, area. The procedure is based on an immunomagnetic separation approach with chlamydial LPS-specific mAbs, followed by DNA purification and total DNA amplification, and subsequent Illumina-based sequence analysis. Quality of genome sequencing was independent of the total number of inclusion-forming units determined for the sample and the amount of non-chlamydial DNA in the Illumina libraries. A geographically and temporally linked clade of isolates was identified with evidence of several different regions of recombination and variable ompA sequence types, suggesting that recombination is common within outbreaks. Culture-independent sequence analysis revealed a linkage pattern at two nucleotide positions that was unique to the genomes of isolates from patients, but not in C. trachomatis recombinants generated in vitro. These data demonstrated that culture-independent sequence analysis can be used to rapidly and inexpensively collect genome data from patients infected by C. trachomatis, and that this approach can be used to examine genomic variation within this species.

  13. Microbial diversity and dynamics throughout manufacturing and ripening of surface ripened semi-hard Danish Danbo cheeses investigated by culture-independent techniques.

    Science.gov (United States)

    Ryssel, Mia; Johansen, Pernille; Al-Soud, Waleed Abu; Sørensen, Søren; Arneborg, Nils; Jespersen, Lene

    2015-12-23

    Microbial successions on the surface and in the interior of surface ripened semi-hard Danish Danbo cheeses were investigated by culture-dependent and -independent techniques. Culture-independent detection of microorganisms was obtained by denaturing gradient gel electrophoresis (DGGE) and pyrosequencing, using amplicons of 16S and 26S rRNA genes for prokaryotes and eukaryotes, respectively. With minor exceptions, the results from the culture-independent analyses correlated to the culture-dependent plating results. Even though the predominant microorganisms detected with the two culture-independent techniques correlated, a higher number of genera were detected by pyrosequencing compared to DGGE. Additionally, minor parts of the microbiota, i.e. comprising cheeses diverged. During cheese production pyrosequencing determined Lactococcus as the dominating genus on cheese surfaces, representing on average 94.7%±2.1% of the OTUs. At day 6 Lactococcus spp. declined to 10.0% of the OTUs, whereas Staphylococcus spp. went from 0.0% during cheese production to 75.5% of the OTUs at smearing. During ripening, i.e. from 4 to 18 weeks, Corynebacterium was the dominant genus on the cheese surface (55.1%±9.8% of the OTUs), with Staphylococcus (17.9%±11.2% of the OTUs) and Brevibacterium (10.4%±8.3% of the OTUs) being the second and third most abundant genera. Other detected bacterial genera included Clostridiisalibacter (5.0%±4.0% of the OTUs), as well as Pseudoclavibacter, Alkalibacterium and Marinilactibacillus, which represented cheeses was dominated by Lactococcus spp. comprising on average 93.9%±7.8% of the OTUs throughout the cheese processing. The microbial dynamics described at genus level in this study add to a comprehensive understanding of the complex microbiota existing especially on surface ripened semi-hard cheeses.

  14. Cultivation-independent methods reveal differences among bacterial gut microbiota in triatomine vectors of Chagas disease.

    Directory of Open Access Journals (Sweden)

    Fabio Faria da Mota

    Full Text Available BACKGROUND: Chagas disease is a trypanosomiasis whose agent is the protozoan parasite Trypanosoma cruzi, which is transmitted to humans by hematophagous bugs known as triatomines. Even though insecticide treatments allow effective control of these bugs in most Latin American countries where Chagas disease is endemic, the disease still affects a large proportion of the population of South America. The features of the disease in humans have been extensively studied, and the genome of the parasite has been sequenced, but no effective drug is yet available to treat Chagas disease. The digestive tract of the insect vectors in which T. cruzi develops has been much less well investigated than blood from its human hosts and constitutes a dynamic environment with very different conditions. Thus, we investigated the composition of the predominant bacterial species of the microbiota in insect vectors from Rhodnius, Triatoma, Panstrongylus and Dipetalogaster genera. METHODOLOGY/PRINCIPAL FINDINGS: Microbiota of triatomine guts were investigated using cultivation-independent methods, i.e., phylogenetic analysis of 16s rDNA using denaturing gradient gel electrophoresis (DGGE and cloned-based sequencing. The Chao index showed that the diversity of bacterial species in triatomine guts is low, comprising fewer than 20 predominant species, and that these species vary between insect species. The analyses showed that Serratia predominates in Rhodnius, Arsenophonus predominates in Triatoma and Panstrongylus, while Candidatus Rohrkolberia predominates in Dipetalogaster. CONCLUSIONS/SIGNIFICANCE: The microbiota of triatomine guts represents one of the factors that may interfere with T. cruzi transmission and virulence in humans. The knowledge of its composition according to insect species is important for designing measures of biological control for T. cruzi. We found that the predominant species of the bacterial microbiota in triatomines form a group of low

  15. Cultivation-Independent Methods Reveal Differences among Bacterial Gut Microbiota in Triatomine Vectors of Chagas Disease

    Science.gov (United States)

    da Mota, Fabio Faria; Marinho, Lourena Pinheiro; Moreira, Carlos José de Carvalho; Lima, Marli Maria; Mello, Cícero Brasileiro; Garcia, Eloi Souza; Carels, Nicolas; Azambuja, Patricia

    2012-01-01

    Background Chagas disease is a trypanosomiasis whose agent is the protozoan parasite Trypanosoma cruzi, which is transmitted to humans by hematophagous bugs known as triatomines. Even though insecticide treatments allow effective control of these bugs in most Latin American countries where Chagas disease is endemic, the disease still affects a large proportion of the population of South America. The features of the disease in humans have been extensively studied, and the genome of the parasite has been sequenced, but no effective drug is yet available to treat Chagas disease. The digestive tract of the insect vectors in which T. cruzi develops has been much less well investigated than blood from its human hosts and constitutes a dynamic environment with very different conditions. Thus, we investigated the composition of the predominant bacterial species of the microbiota in insect vectors from Rhodnius, Triatoma, Panstrongylus and Dipetalogaster genera. Methodology/Principal Findings Microbiota of triatomine guts were investigated using cultivation-independent methods, i.e., phylogenetic analysis of 16s rDNA using denaturing gradient gel electrophoresis (DGGE) and cloned-based sequencing. The Chao index showed that the diversity of bacterial species in triatomine guts is low, comprising fewer than 20 predominant species, and that these species vary between insect species. The analyses showed that Serratia predominates in Rhodnius, Arsenophonus predominates in Triatoma and Panstrongylus, while Candidatus Rohrkolberia predominates in Dipetalogaster. Conclusions/Significance The microbiota of triatomine guts represents one of the factors that may interfere with T. cruzi transmission and virulence in humans. The knowledge of its composition according to insect species is important for designing measures of biological control for T. cruzi. We found that the predominant species of the bacterial microbiota in triatomines form a group of low complexity whose structure

  16. Monitoring the ecology of Bacillus during Daqu incubation, a fermentation starter, using culture-dependent and culture-independent methods

    NARCIS (Netherlands)

    Zheng, Y.; Zheng, X.; Han, B.Z.; Han, J.S.; Nout, M.J.R.; Chen, J.Y.

    2013-01-01

    Daqu, a traditional fermentation starter, has been used to produce attractively flavored foods such as vinegar and Chinese liquor for thousands of years. Although Bacillus spp. are one of the dominant microorganisms in Daqu, more precise information is needed to reveal why and how Bacillus became do

  17. Micro-seismic Imaging Using a Source Independent Waveform Inversion Method

    KAUST Repository

    Wang, Hanchen

    2016-04-18

    Micro-seismology is attracting more and more attention in the exploration seismology community. The main goal in micro-seismic imaging is to find the source location and the ignition time in order to track the fracture expansion, which will help engineers monitor the reservoirs. Conventional imaging methods work fine in this field but there are many limitations such as manual picking, incorrect migration velocity and low signal to noise ratio (S/N). In traditional surface survey imaging, full waveform inversion (FWI) is widely used. The FWI method updates the velocity model by minimizing the misfit between the observed data and the predicted data. Using FWI to locate and image microseismic events allows for an automatic process (free of picking) that utilizes the full wavefield. Use the FWI technique, and overcomes the difficulties of manual pickings and incorrect velocity model for migration. However, the technique of waveform inversion of micro-seismic events faces its own problems. There is significant nonlinearity due to the unknown source location (space) and function (time). We have developed a source independent FWI of micro-seismic events to simultaneously invert for the source image, source function and velocity model. It is based on convolving reference traces with the observed and modeled data to mitigate the effect of an unknown source ignition time. The adjoint-state method is used to derive the gradient for the source image, source function and velocity updates. To examine the accuracy of the inverted source image and velocity model the extended image for source wavelet in z-axis is extracted. Also the angle gather is calculated to check the applicability of the migration velocity. By inverting for the source image, source wavelet and the velocity model simultaneously, the proposed method produces good estimates of the source location, ignition time and the background velocity in the synthetic experiments with both parts of the Marmousi and the SEG

  18. Ethos of Independence across Regions in the United States: The Production-Adoption Model of Cultural Change

    Science.gov (United States)

    Kitayama, Shinobu; Conway, Lucian Gideon, III; Pietromonaco, Paula R.; Park, Hyekyung; Plaut, Victoria C.

    2010-01-01

    Contemporary U.S. culture has a highly individualistic ethos. Nevertheless, exactly how this ethos was historically fostered remains unanalyzed. A new model of dynamic cultural change maintains that sparsely populated, novel environments that impose major threats to survival, such as the Western frontier in the United States during the 18th and…

  19. How Can We Overcome the Dichotomy That Western Culture Has Created between the Concepts of Independence and Dependence?

    Science.gov (United States)

    Gross, Zehavit

    2015-01-01

    The purpose of this article, inspired by the works of Martin Buber, is to propose an alternative to the inherent dichotomy of Western culture. It may allow Western culture to transcend its fixed nature towards new directions and to suggest challenging solutions for reshaping the questions--what is the role of man in the world, and what is the…

  20. Rapid culture-independent microbial analysis aboard the international space station (ISS) stage two: quantifying three microbial biomarkers.

    Science.gov (United States)

    Morris, Heather C; Damon, Michael; Maule, Jake; Monaco, Lisa A; Wainwright, Norm

    2012-09-01

    Abstract A portable, rapid, microbial detection unit, the Lab-On-a-Chip Application Development Portable Test System (LOCAD-PTS), was launched to the International Space Station (ISS) as a technology demonstration unit in December 2006. Results from the first series of experiments designed to detect Gram-negative bacteria on ISS surfaces by quantifying a single microbial biomarker lipopolysaccharide (LPS) were reported in a previous article. Herein, we report additional technology demonstration experiments expanding the on-orbit capabilities of the LOCAD-PTS to detecting three different microbial biomarkers on ISS surfaces. Six different astronauts on more than 20 occasions participated in these experiments, which were designed to test the new beta-glucan (fungal cell wall molecule) and lipoteichoic acid (LTA; Gram-positive bacterial cell wall component) cartridges individually and in tandem with the existing Limulus Amebocyte Lysate (LAL; Gram-negative bacterial LPS detection) cartridges. Additionally, we conducted the sampling side by side with the standard culture-based detection method currently used on the ISS. Therefore, we present data on the distribution of three microbial biomarkers collected from various surfaces in every module present on the ISS at the time of sampling. In accordance with our previous experiments, we determined that spacecraft surfaces known to be frequently in contact with crew members demonstrated higher values of all three microbial molecules. Key Words: Planetary protection-Spaceflight-Microbiology-Biosensor. Astrobiology 12, 830-840.

  1. A NEW METHOD FOR SAFETY CULTURE EVALUATION IN PROCESS SYSTEMS

    OpenAIRE

    Carvalho,Paulo Victor Rodrigues de; VIDAL, Mario Cesar Rodríguez; GRECCO, Cláudio Henrique dos Santos; Cosenza, Carlos Alberto Nunes

    2014-01-01

    The culture of any organization that deals with safe-critical processes should be centered on safety. Safety culture reflects human awareness of the significant destructive capability of process plants when control is lost, and the recognition that strict attention to safety is essential to achieve the expected benefits. Safety culture appeared as paradigmatic change in the safety approach for process plants in the aftermath of the Chernobyl and Bhopal accidents. The Internati...

  2. Antibiotics and antibiotic resistance in agroecosystems: Cultural methods and gaps in knowledge

    Science.gov (United States)

    Varying cultural methodologies are used in assessment of antibiotic resistance in environmental samples. Culture based methods commonly involve isolation of target bacteria on general or selective media, and assessing growth in response to specific concentrations of antibiotics. Though time consumin...

  3. Utilizing the Project Method for Teaching Culture and Intercultural Competence

    Science.gov (United States)

    Euler, Sasha S.

    2017-01-01

    This article presents a detailed methodological outline for teaching culture through project work. It is argued that because project work makes it possible to gain transferrable and applicable knowledge and insight, it is the ideal tool for teaching culture with the aim of achieving real intercultural communicative competence (ICC). Preceding the…

  4. A novel 2.5D culture platform to investigate the role of stiffness gradients on adhesion-independent cell migration.

    Directory of Open Access Journals (Sweden)

    Mark-Phillip Pebworth

    Full Text Available Current studies investigating the role of biophysical cues on cell migration focus on the use of culture platforms with static material parameters. However, migrating cells in vivo often encounter spatial variations in extracellular matrix stiffness. To better understand the effects of stiffness gradients on cell migration, we developed a 2.5D cell culture platform where cells are sandwiched between stiff tissue culture plastic and soft alginate hydrogel. Under these conditions, we observed migration of cells from the underlying stiff substrate into the alginate matrix. Observation of migration into alginate in the presence of integrin inhibition as well as qualitative microscopic analyses suggested an adhesion-independent cell migration mode. Observed migration was dependent on alginate matrix stiffness and the RhoA-ROCK-myosin-II pathway; inhibitors specifically targeting ROCK and myosin-II arrested cell migration. Collectively, these results demonstrate the utility of the 2.5D culture platform to advance our understanding of the effects of stiffness gradients and mechanotransductive signaling on adhesion-independent cell migration.

  5. S-adenosyl-methionine decreases ethanol-induced apoptosis in primary hepatocyte cultures by a c-Jun N-terminal kinase activity-independent mechanism

    Institute of Scientific and Technical Information of China (English)

    María del Pilar Cabrales-Romero; Lucrecia Márquez-Rosado; Samia Fattel-Fazenda; Cristina Trejo-Solís; Evelia Arce-Popoca; Leticia Alemén-Lazarini; Saúl Villa-Trevi(n)o

    2006-01-01

    AIM: To determine the role of c-Jun N-terminal kinase (JNK) activity in ethanol-induced apoptosis and the modulation of this signaling cascade by S-Adenosylmethionine (AdoMet).METHODS: Primary hepatocyte cultures were pretreated with 100 μmol/L SP600125, a selective JNK inhibitor, 1 mL/L DMSO or 4 mmol/L AdoMet and then exposed to 100 mmo/L ethanol. Hepatocyte apoptosis was determined by the TUNEL and DNA ladder assays.JNK activity and its inhibition by SP600125 and AdoMet were determined by Western blot analysis of c-jun phosphorylation and Bid fragmentation. SP600125 and AdoMet effects on the apoptotic signaling pathway were determined by Western blot analysis of cytochrome c release and pro-caspase 3 fragmentation. The AdoMet effect on glutathione levels was measured by Ellman's method and reactive oxygen species (ROS) generation by cell cytometry.RESULTS: The exposure of hepatocytes to ethanol induced JNK activation, c-jun phosphorylation, Bid fragmentation, cytochrome c release and pro-caspase 3 cleavage; these effects were diminished by SP600125, and caused a significant decreasein ethanol-induced apoptosis (P< 0.05). AdoMet exerted an antioxidant effect maintaining glutathione levels and decreasing ROS generation, without a significant effect on JNK activity,and prevented cytochrome c release and pro-caspase 3 cleavage.CONCLUSION: The JNK signaling cascade is a key component of the proapoptotic signaling pathway induced by ethanol. JNK activation may be independent from ROS generation, since AdoMet which exerted antioxidant properties did not have a significant effect on JNK activity. JNK pathway modulator agents and AdoMet may be components of promising therapies for alcoholic liver disease (ALD) treatment.

  6. Correlations between social-emotional feelings and anterior insula activity are independent from visceral states but influenced by culture

    Directory of Open Access Journals (Sweden)

    Mary Helen eImmordino-Yang

    2014-09-01

    Full Text Available The anterior insula (AI maps visceral states and is active during emotional experiences, a functional confluence that is central to neurobiological accounts of feelings. Yet, it is unclear how AI activity correlates with feelings during social emotions, and whether this correlation may be influenced by culture, as studies correlating real-time AI activity with visceral states and feelings have focused on Western subjects feeling physical pain or basic disgust. Given psychological evidence that social-emotional feelings are cognitively constructed within cultural frames, we asked Chinese and American participants to report their feeling strength to admiration and compassion-inducing narratives during fMRI with simultaneous electrocardiogram recording. Trial-by-trial, cardiac arousal and feeling strength correlated with ventral and dorsal AI activity bilaterally but predicted different variance, suggesting that interoception and social-emotional feeling construction are concurrent but dissociable AI functions. Further, although the variance that correlated with cardiac arousal did not show cultural effects, the variance that correlated with feelings did. Feeling strength was especially associated with ventral AI activity (the autonomic modulatory sector in the Chinese group but with dorsal AI activity (the visceral-somatosensory/cognitive sector in an American group not of Asian descent. This cultural group difference held after controlling for posterior insula activity and was replicated. A bi-cultural East-Asian American group showed intermediate results. The findings help elucidate how the AI supports feelings and suggest that previous reports that dorsal AI activation reflects feeling strength are culture related. More broadly, the results suggest that the brain’s ability to construct conscious experiences of social emotion is less closely tied to visceral processes than neurobiological models predict and at least partly open to cultural

  7. Adaptação transcultural da Escala de Independência em Atividades da Vida Diária (Escala de Katz Cross-cultural adaptation of the Independence in Activities of Daily Living Index (Katz Index

    Directory of Open Access Journals (Sweden)

    Valéria Teresa Saraiva Lino

    2008-01-01

    Full Text Available Desenvolvimento e análise do desempenho de uma adaptação transcultural para o português da Escala de Katz de independência em atividades da vida diária. Duas traduções e duas retrotraduções analisaram as equivalências conceitual, de itens e semântica para a escolha da versão final. A equivalência operacional foi avaliada em um estudo piloto, testando-se a confiabilidade e a consistência interna da versão adaptada por meio de reteste no mesmo dia em 156 pacientes ou após sete dias da primeira entrevista. A resolução de diferenças sutis em alguns itens levou à equivalência cultural. A versão final foi considerada fácil de se entender e de aplicar. A concordância corrigida para o acaso (kappa ponderado foi de 0,91. O alfa de Chronbach variou de 0,80 a 0,92. A versão em português da Escala de Katz de independência em atividades da vida diária, completamente desenvolvida e testada, provou ser equivalente à original em inglês. Os itens apresentaram consistência interna e as taxa foram confiáveis.This study involved the development and performance assessment of a cultural adaptation of the Katz scale of independence in activities of daily living, translated into Portuguese in Brazil. Two translations and two back-translations of the items were analyzed by experts in order to decide on the final version. Operational equivalence was assessed in a pilot study. The reliability and internal consistency of the adapted version were assessed by retesting 156 patients on the same day or 7 days after the first interview. Cultural equivalence was achieved after resolving subtle differences in some items. The final version was considered easy to understand and use. Chance-corrected agreement (weighted kappa was 0.91. Cronbach's alpha ranged from 0.80 to 0.92. CONCLUSIONS: a Portuguese version of the Katz scale of independence in activities of daily living, thoroughly developed and tested, proved equivalent to the original version

  8. Transnational Cultural Leadership as a Situated Practice : Dilemmas and Methods

    NARCIS (Netherlands)

    Kolsteeg, Johan

    2017-01-01

    In a longitudinal transnational research project, a network of European research institutions and field organisations aims to understand how cultural managers mediate global and local pressures concerning creative autonomy, economy and ideology. Among the research questions are which variables

  9. Monitoring of changes in lactic acid bacteria during production of Thai traditional fermented shrimp (Kung-Som) by culturing method and PCR-DGGE technique

    OpenAIRE

    Kanokwan Thongruck; Sutanate Saelao; Punnanee Sumpavapol; Soottawat Benjakul; Suppasil Maneerat

    2017-01-01

    The lactic acid bacteria (LAB) ecology of Kung-Som, a traditional Thai fermented shrimp, was investigated using culture-dependent and culture-independent methods. Kung-Som was fermented at room temperature under anaerobic condition for 7 days using autochthonous fermentation. The viable counts revealed a dominance of LAB, from the 16S rRNA gene V6-V8 sequence analysis of the isolates, Enterococcus faecalis, Enterococcus sanguinicola, Weissella cibaria, Weissella confusa, Enterococ...

  10. Cell contact as an independent factor modulating cardiac myocyte hypertrophy and survival in long-term primary culture

    Science.gov (United States)

    Clark, W. A.; Decker, M. L.; Behnke-Barclay, M.; Janes, D. M.; Decker, R. S.

    1998-01-01

    Cardiac myocytes maintained in cell culture develop hypertrophy both in response to mechanical loading as well as to receptor-mediated signaling mechanisms. However, it has been shown that the hypertrophic response to these stimuli may be modulated through effects of intercellular contact achieved by maintaining cells at different plating densities. In this study, we show that the myocyte plating density affects not only the hypertrophic response and features of the differentiated phenotype of isolated adult myocytes, but also plays a significant role influencing myocyte survival in vitro. The native rod-shaped phenotype of freshly isolated adult myocytes persists in an environment which minimizes myocyte attachment and spreading on the substratum. However, these conditions are not optimal for long-term maintenance of cultured adult cardiac myocytes. Conditions which promote myocyte attachment and spreading on the substratum, on the other hand, also promote the re-establishment of new intercellular contacts between myocytes. These contacts appear to play a significant role in the development of spontaneous activity, which enhances the redevelopment of highly differentiated contractile, junctional, and sarcoplasmic reticulum structures in the cultured adult cardiomyocyte. Although it has previously been shown that adult cardiac myocytes are typically quiescent in culture, the addition of beta-adrenergic agonists stimulates beating and myocyte hypertrophy, and thereby serves to increase the level of intercellular contact as well. However, in densely-plated cultures with intrinsically high levels of intercellular contact, spontaneous contractile activity develops without the addition of beta-adrenergic agonists. In this study, we compare the function, morphology, and natural history of adult feline cardiomyocytes which have been maintained in cultures with different levels of intercellular contact, with and without the addition of beta-adrenergic agonists

  11. The projection Galerkin method for solving the time-independent differential diffusion equation in a semi-infinite domain

    Science.gov (United States)

    Makarenkov, A. M.; Seregina, E. V.; Stepovich, M. A.

    2017-05-01

    Using the diffusion equation as an example, results of applying the projection Galerkin method for solving time-independent heat and mass transfer equations in a semi-infinite domain are presented. The convergence of the residual corresponding to the approximate solution of the timeindependent diffusion equation obtained by the projection method using the modified Laguerre functions is proved. Computational results for a two-dimensional toy problem are presented.

  12. A novel method for generating xeno-free human feeder cells for human embryonic stem cell culture.

    Science.gov (United States)

    Meng, Guoliang; Liu, Shiying; Krawetz, Roman; Chan, Michael; Chernos, Judy; Rancourt, Derrick E

    2008-06-01

    Long-term cultures of human embryonic stem (hES) cells require a feeder layer for maintaining cells in an undifferentiated state and increasing karyotype stability. In routine hES cell culture, mouse embryonic fibroblast (MEF) feeders and animal component-containing media (FBS or serum replacement) are commonly used. However, the use of animal materials increases the risk of transmitting pathogens to hES cells and therefore is not optimal for use in cultures intended for human transplantation. There are other limitations with conventional feeder cells, such as MEFs, which have a short lifespan and can only be propagated five to six passages before senescing. Several groups have investigated maintaining existing hES cell lines and deriving new hES cell lines on human feeder layers. However, almost all of these human source feeder cells employed in previous studies were derived and cultured in animal component conditions. Even though one group previously reported the derivation and culture of human foreskin fibroblasts (HFFs) in human serum-containing medium, this medium is not optimal because HFFs routinely undergo senescence after 10 passages when cultured in human serum. In this study we have developed a completely animal-free method to derive HFFs from primary tissues. We demonstrate that animal-free (AF) HFFs do not enter senescence within 55 passages when cultured in animal-free conditions. This methodology offers alternative and completely animal-free conditions for hES cell culture, thus maintaining hES cell morphology, pluripotency, karyotype stability, and expression of pluripotency markers. Moreover, no difference in hES cell maintenance was observed when they were cultured on AF-HFFs of different passage number or independent derivations.

  13. Organizational Culture and the Deployment of Agile Methods: The Competing Values Model View

    Science.gov (United States)

    Iivari, Juhani; Iivari, Netta

    A number of researchers have identified organizational culture as a factor that potentially affects the deployment of agile systems development methods. Inspired by the study of Iivari and Huisman (2007), which focused on the deployment of traditional systems development methods, the present paper proposes a number of hypotheses about the influence of organizational culture on the deployment of agile methods.

  14. A novel three primers PCR (TP-PCR) method to obtain recombinant DNA molecule independent of restriction enzyme

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    In this note, we report a novel and efficient three primers PCR (TP-PCR) method to rapidly generate recombinant DNA molecule at precise junction between two arbitrary DNA fragments. TP-PCR method is characterized by its reaction system with two templates and three primers, which can produce a recombinant DNA molecule in one PCR reaction. The main advantages of this method are the independence of sequences at the recombination site, the rapidness, and the easy establishment of adequate conditions. This method has been successfully applied to constructing a fusion protein gene, sck gene.

  15. Methods for SWATH™: Data Independent Acquisition on TripleTOF Mass Spectrometers.

    Science.gov (United States)

    Holewinski, Ronald J; Parker, Sarah J; Matlock, Andrea D; Venkatraman, Vidya; Van Eyk, Jennifer E

    2016-01-01

    Data independent acquisition (DIA also termed SWATH) is an emerging technology in the field of mass spectrometry based proteomics. Although the concept of DIA has been around for over a decade, the recent advancements, in particular the speed of acquisition, of mass analyzers have pushed the technique into the spotlight and allowed for high-quality DIA data to be routinely acquired by proteomics labs. In this chapter we will discuss the protocols used for DIA acquisition using the Sciex TripleTOF mass spectrometers and data analysis using the Sciex processing software.

  16. Flow cytometric methods to investigate culture heterogeneities for plant metabolic engineering.

    Science.gov (United States)

    Gaurav, Vishal; Kolewe, Martin E; Roberts, Susan C

    2010-01-01

    Plant cell cultures provide an important method for production and supply of a variety of natural products, where conditions can be easily controlled, manipulated, and optimized. Development and optimization of plant cell culture processes require both bioprocess engineering and metabolic engineering approaches. Cultures are generally highly heterogeneous, with significant variability amongst cells in terms of growth, metabolism, and productivity of key metabolites. Taxus cultures produce the important anti-cancer agent Taxol((R)) (i.e., paclitaxel) and have demonstrated significant variability amongst cell populations in culture with regard to paclitaxel accumulation, cell cycle participation, and protein synthesis. To fully understand the link between cellular metabolism and culture behavior and to enable targeted metabolic engineering approaches, cultures need to be studied at a single cell level. This chapter describes the application of plant cell flow cytometric techniques to investigate culture heterogeneity at the single cell level, in order to optimize culture performance through targeted metabolic engineering. Flow cytometric analytical methods are described to study Taxus single cells, protoplasts, and nuclei suspensions with respect to secondary metabolite accumulation, DNA content, cell size, and complexity. Reproducible methods to isolate these single particle suspensions from aggregated Taxus cultures are discussed. Methods to stain both fixed and live cells for a variety of biological markers are provided to enable characterization of cell phenotypes. Fluorescence-activated cell sorting (FACS) methods are also presented to facilitate isolation of certain plant cell culture populations for both analysis and propagation of superior cell lines for use in bioprocesses.

  17. A subject-independent method for automatically grading electromyographic features during a fatiguing contraction.

    Science.gov (United States)

    Chattopadhyay, Rita; Jesunathadas, Mark; Poston, Brach; Santello, Marco; Ye, Jieping; Panchanathan, Sethuraman

    2012-06-01

    Many studies have attempted to monitor fatigue from electromyogram (EMG) signals. However, fatigue affects EMG in a subject-specific manner. We present here a subject-independent framework for monitoring the changes in EMG features that accompany muscle fatigue based on principal component analysis and factor analysis. The proposed framework is based on several time- and frequency-domain features, unlike most of the existing work, which is based on two to three features. Results show that latent factors obtained from factor analysis on these features provide a robust and unified framework. This framework learns a model from EMG signals of multiple subjects, that form a reference group, and monitors the changes in EMG features during a sustained submaximal contraction on a test subject on a scale from zero to one. The framework was tested on EMG signals collected from 12 muscles of eight healthy subjects. The distribution of factor scores of the test subject, when mapped onto the framework was similar for both the subject-specific and subject-independent cases.

  18. A comparison on parameter-estimation methods in multiple regression analysis with existence of multicollinearity among independent variables

    Directory of Open Access Journals (Sweden)

    Hukharnsusatrue, A.

    2005-11-01

    Full Text Available The objective of this research is to compare multiple regression coefficients estimating methods with existence of multicollinearity among independent variables. The estimation methods are Ordinary Least Squares method (OLS, Restricted Least Squares method (RLS, Restricted Ridge Regression method (RRR and Restricted Liu method (RL when restrictions are true and restrictions are not true. The study used the Monte Carlo Simulation method. The experiment was repeated 1,000 times under each situation. The analyzed results of the data are demonstrated as follows. CASE 1: The restrictions are true. In all cases, RRR and RL methods have a smaller Average Mean Square Error (AMSE than OLS and RLS method, respectively. RRR method provides the smallest AMSE when the level of correlations is high and also provides the smallest AMSE for all level of correlations and all sample sizes when standard deviation is equal to 5. However, RL method provides the smallest AMSE when the level of correlations is low and middle, except in the case of standard deviation equal to 3, small sample sizes, RRR method provides the smallest AMSE.The AMSE varies with, most to least, respectively, level of correlations, standard deviation and number of independent variables but inversely with to sample size.CASE 2: The restrictions are not true.In all cases, RRR method provides the smallest AMSE, except in the case of standard deviation equal to 1 and error of restrictions equal to 5%, OLS method provides the smallest AMSE when the level of correlations is low or median and there is a large sample size, but the small sample sizes, RL method provides the smallest AMSE. In addition, when error of restrictions is increased, OLS method provides the smallest AMSE for all level, of correlations and all sample sizes, except when the level of correlations is high and sample sizes small. Moreover, the case OLS method provides the smallest AMSE, the most RLS method has a smaller AMSE than

  19. Development of an Evaluation Method for Team Safety Culture Competencies using Social Network Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Han, Sang Min; Kim, Ar Ryum; Seong, Poong Hyun [KAIST, Daejeon (Korea, Republic of)

    2016-05-15

    In this study, team safety culture competency of a team was estimated through SNA, as a team safety culture index. To overcome the limit of existing safety culture evaluation methods, the concept of competency and SNA were adopted. To estimate team safety culture competency, we defined the definition, range and goal of team safety culture competencies. Derivation of core team safety culture competencies is performed and its behavioral characteristics were derived for each safety culture competency, from the procedures used in NPPs and existing criteria to assess safety culture. Then observation was chosen as a method to provide the input data for the SNA matrix of team members versus insufficient team safety culture competencies. Then through matrix operation, the matrix was converted into the two meaningful values, which are density of team members and degree centralities of each team safety culture competency. Density of tem members and degree centrality of each team safety culture competency represent the team safety culture index and the priority of team safety culture competency to be improved.

  20. Possibilities of the Integration of the Method of the Ecologically Oriented Independent Scientific Research in the Study Process

    Science.gov (United States)

    Grizans, Jurijs; Vanags, Janis

    2010-01-01

    The aim of this paper is to analyse possibilities of the integration of the method of the ecologically oriented independent scientific research in the study process. In order to achieve the set aim, the following scientific research methods were used: analysis of the conceptual guidelines for the development of environmentally oriented entrepreneurship, interpretation of the experts' evaluation of the ecologically oriented management, analysis of the results of the students' ecologically oriented independent scientific research, as well as monographic and logically constructive methods. The results of the study give an opportunity to make conclusions and to develop conceptual recommendations on how to introduce future economics and business professionals with the theoretical and practical aspects of ecologically oriented management during the study process.

  1. Polyphasic Approach to Bacterial Dynamics during the Ripening of Spanish Farmhouse Cheese, Using Culture-Dependent and -Independent Methods▿

    OpenAIRE

    Martín-Platero, Antonio M.; Valdivia, Eva; Maqueda, Mercedes; Martín-Sánchez,Inés; Martínez-Bueno, Manuel

    2008-01-01

    We studied the dynamics of the microbial population during ripening of Cueva de la Magahá cheese using a combination of classical and molecular techniques. Samples taken during ripening of this Spanish goat's milk cheese in which Lactococcus lactis and Streptococcus thermophilus were used as starter cultures were analyzed. All bacterial isolates were clustered by using randomly amplified polymorphic DNA (RAPD) and identified by 16S rRNA gene sequencing, species-specific PCR, and multiplex PCR...

  2. Teaching Geographic Field Methods to Cultural Resource Management Technicians

    Science.gov (United States)

    Mires, Peter B.

    2004-01-01

    There are perhaps 10,000 technicians in the United States who work in the field known as cultural resource management (CRM). The typical field technician possesses a bachelor's degree in anthropology, geography, or a closely allied discipline. The author's experience has been that few CRM field technicians receive adequate undergraduate training…

  3. A Stock Market Prediction Method Based on Support Vector Machines (SVM and Independent Component Analysis (ICA

    Directory of Open Access Journals (Sweden)

    Hakob GRIGORYAN

    2016-08-01

    Full Text Available The research presented in this work focuses on financial time series prediction problem. The integrated prediction model based on support vector machines (SVM with independent component analysis (ICA (called SVM-ICA is proposed for stock market prediction. The presented approach first uses ICA technique to extract important features from the research data, and then applies SVM technique to perform time series prediction. The results obtained from the SVM-ICA technique are compared with the results of SVM-based model without using any pre-processing step. In order to show the effectiveness of the proposed methodology, two different research data are used as illustrative examples. In experiments, the root mean square error (RMSE measure is used to evaluate the performance of proposed models. The comparative analysis leads to the conclusion that the proposed SVM-ICA model outperforms the simple SVM-based model in forecasting task of nonstationary time series.

  4. Grain size evaluation of structural materials in nuclear power plant using a thickness independent ultrasonic method

    Energy Technology Data Exchange (ETDEWEB)

    Li, Xiongbing, E-mail: lixb_ex@163.com [CAD/CAM Institute, Central South University, Changsha 410075 (China); State Key Laboratory of Powder Metallurgy, Central South University, Changsha 410083 (China); Song, Yongfeng [CAD/CAM Institute, Central South University, Changsha 410075 (China); Ni, Peijun [The Ningbo Branch of Ordnance Science Institute of China, Ningbo 315103 (China); Wang, Zi; Liu, Feng [State Key Laboratory of Powder Metallurgy, Central South University, Changsha 410083 (China); Du, Hualong [Mechanical and Aerospace Engineering Department, North Carolina State University, Raleigh, NC 27606 (United States)

    2015-05-15

    Highlights: •We derive a coefficient of ultrasonic attenuation rate related to the grain size. •The mean grain size of the pipe can be evaluated without measuring its thickness. •Experiments show this method is better suited to square pipe than other methods. -- Abstract: It is important to accurately and nondestructively evaluate the grain size of structural materials used in nuclear power plants. The current ultrasonic non-destructive methods are so dependent on the thickness measurement of a square pipe that it reduces their practicality and reliability. In this paper, a novel method using the coefficient of ultrasonic attenuation rate is developed by using the transmission and reflection coefficients. As a result, the mean grain size of the pipe can be nondestructively evaluated without measuring its thickness. Moreover, the signal preprocessing is studied to improve the stability and accuracy of evaluation results. The experimental results show that the dependence of the attenuation rate on grain sizes is much higher than that of the ultrasonic velocity. The relative error of the attenuation rate method is lower than that of the backscatter method if the thickness of the sample is less than 5 mm. When evaluating a TP304 stainless steel square pipe whose thickness is not convenient to measure, the mean grain sizes are measured 103.5 ± 2.6 μm, 96.9 ± 3.5 μm and 94.0 ± 1.7 μm by the attenuation method, the attenuation rate method and the electron backscattering diffraction method, respectively. The result verifies that the presented method works better than the attenuation method due to the fact that the error of the thickness measurement has no effect on the ultrasonic attenuation rate.

  5. A method for segmentation and motion estimation of multiple independently moving objects

    NARCIS (Netherlands)

    Willemink, G.H.; Heijden, van der F.

    2006-01-01

    The aim of this work is to design a robust method for online estimation of object motion and structure in a dynamic scene. A method for segmentation and estimation of object motion and structure from 3-d points in a dynamic scene, observed by a sequence of stereo images, is proposed. The proposed me

  6. Novel MAPK-dependent and -independent tubulogenes identified via microarray analysis of 3D-cultured Madin-Darby canine kidney cells.

    Science.gov (United States)

    Chacon-Heszele, Maria F; Zuo, Xiaofeng; Hellman, Nathan E; McKenna, Sarah; Choi, Soo Young; Huang, Liwei; Tobias, John W; Park, Kwon Moo; Lipschutz, Joshua H

    2014-05-01

    Cystogenesis and tubulogenesis are basic building blocks for many epithelial organs, including the kidney. Most researchers have used two-dimensional (2D) cell culture to investigate signaling pathways downstream of hepatocyte growth factor (HGF). We hypothesize that three-dimensional (3D) collagen-grown Madin-Darby canine kidney (MDCK) cells, which form cysts and then tubulate in response to HGF, are a much more in vivo-like system for the identification of novel tubulogenes. With the use of a canine microarray containing over 20,000 genes, 2,417 genes were identified as potential tubulogenes that were differentially regulated, exclusively in 3D-grown MDCK cells. Among these, 840 were dependent on MAPK signaling. Importantly, this work shows that many putative tubulogenes, previously identified via microarray analysis of 2D cultures, including by us, do not change in 3D culture and vice versa. The use of a 3D-culture system allowed for the identification of novel MAPK-dependent and -independent genes that regulate early renal tubulogenesis in vitro, e.g., matrix metalloproteinase 1 (MMP1). Knockdown of MMP1 led to defects in cystogenesis and tubulogenesis in 3D-grown MDCK cells, most likely due to problems establishing normal polarity. We suggest that data obtained from 2D cultures, even those using MDCK cells treated with HGF, should not be automatically extrapolated to factors important for cystogenesis and tubulogenesis. Instead, 3D culture, which more closely replicates the biological environment and is therefore a more accurate model for identifying tubulogenes, is preferred. Results from the present analysis will be used to build a more accurate model of the signaling pathways that control cystogenesis and tubulogenesis.

  7. Rapid method for identification of group B streptococci in neonatal blood cultures.

    OpenAIRE

    Holmes, R. L.; Harada, W A

    1981-01-01

    A rapid technique used for the identification of Streptococcus agalactiae, Lancefield group B, from the blood cultures of two neonatal infants is reported. The method utilized the Phadebact Streptococcus Test System (Pharmacia Diagnostics, Piscataway, N.J.) and the supernatant from 13- and 14-h blood cultures. Additional studies with simulated neonatal blood cultures revealed that this method was reproducible. Additional studies also revealed that some non-specific agglutination did occur, wh...

  8. Using case methods to study cultural diversity within the development of telematic systems

    DEFF Research Database (Denmark)

    Nøhr, Christian; Bertelsen, Pernille Scholdan; Brender, Jytte

    The report " Using case methods to study cultural diversity within the development of telematic systems" discusses a case study method which is an extension of work orginallly done in Babel, a 5th framework EU project. The report contributes to the discussion identifying operational cultural...

  9. Using case methods to study cultural diversity within the development of telematic systems

    DEFF Research Database (Denmark)

    Nøhr, Christian; Bertelsen, Pernille Scholdan; Brender, Jytte

    The report " Using case methods to study cultural diversity within the development of telematic systems" discusses a case study method which is an extension of work orginallly done in Babel, a 5th framework EU project. The report contributes to the discussion identifying operational cultural...

  10. Cross-Cultural Group Counseling and the Use of the Sentence Completion Method.

    Science.gov (United States)

    Ruiz, Sandoval Aureliano

    1984-01-01

    Describes the use of the sentence completion method in multi-ethnic groups using a set of eight cross-cultural sentence items. Participants' feedback suggested that the method has been instrumental in the exploration and conceptual understanding of cultural, ethnical and lingual aspects. (JAC)

  11. Torque Distribution Algorithm for an Independently Driven Electric Vehicle Using a Fuzzy Control Method

    Directory of Open Access Journals (Sweden)

    Jinhyun Park

    2015-08-01

    Full Text Available The in-wheel electric vehicle is expected to be a popular next-generation vehicle because an in-wheel system can simplify the powertrain and improve driving performance. In addition, it also has an advantage in that it maximizes driving efficiency through independent torque control considering the motor efficiency. However, there is an instability problem if only the driving torque is controlled in consideration of only the motor efficiency. In this paper, integrated torque distribution strategies are proposed to overcome these problems. The control algorithm consists of various strategies for optimizing driving efficiency, satisfying driver demands, and considering tire slip and vehicle cornering. Fuzzy logic is used to determine the appropriate timing of intervention for each distribution strategy. A performance simulator for in-wheel electric vehicles was developed by using MATLAB/Simulink and CarSim to validate the control strategies. From simulation results under complex driving conditions, the proposed algorithm was verified to improve both the driving stability and fuel economy of the in-wheel vehicle.

  12. Polarimetry-based method to extract geometry-independent metrics of tissue anisotropy.

    Science.gov (United States)

    Wallenburg, Marika A; Wood, Michael F G; Ghosh, Nirmalya; Vitkin, I Alex

    2010-08-01

    Recently, we have used polarimetry as a method for assessing the linear retardance of infarcted myocardium. While linear retardance reflects tissue anisotropy, experimental geometry has a confounding effect due to dependence of the linear retardance on the orientation of the sample with respect to the probing beam. Here, polarimetry imaging of an 8mm diameter birefringent polystyrene sphere of known anisotropy axis was used to test a dual-projection method by which the anisotropy axis and its true magnitude can be reconstructed, thus eliminating the confounding effect of anisotropy axis orientation. Feasibility is demonstrated in ex-vivo tissue imaging.

  13. In Search of an Alternative Feminist Cinema: Gender, Crisis, and the Cultural Discourse of Nation Building in Chinese Independent Films

    Directory of Open Access Journals (Sweden)

    Jinhua Li

    2017-04-01

    Full Text Available Chinese feminist cinema in the postsocialist era is shaped by the grand narrative of nation building that glamorizes urban professional career women and their contributions to economic marketization and globalization. Such cinematic overemphasis on urban women proves inadequate as it creates a disturbing silence about the diasporic existence of non-urban women. This uneven condition demands the creation of an alternative cinematic feminism that visualizes the diversity of Chinese women and represents the heterogeneity of feminist cinematic expressions and female experiences. Using Li Yu’s 'Lost in Beijing' (2007, 'Pingguo' 苹果 and Li Yang’s 'Blind Mountain' (2007, 'Mang shan' 盲山 as case studies, this essay investigates how Chinese independent films re-negotiate female gender identity and crisis through commercialized visual realism and social intervention while in reality the postsocialist grand narrative of nation building redefines the living conditions of female migrant workers and women of limited resources.

  14. Similar estimates of temperature impacts on global wheat yield by three independent methods

    DEFF Research Database (Denmark)

    Liu, Bing; Asseng, Senthold; Müller, Christoph

    2016-01-01

    The potential impact of global temperature change on global crop yield has recently been assessed with different methods. Here we show that grid-based and point-based simulations and statistical regressions (from historic records), without deliberate adaptation or CO2 fertilization effects, produ...

  15. [Limb-length measurements using wooden boards: an accurate and experience-independent method

    NARCIS (Netherlands)

    Pakvis, D.F.M.; Jaarsma, R.L.; Kampen, A. van

    2003-01-01

    OBJECTIVE: To determine the precision and reliability of the indirect limb-length measurement, and the inter-observer variance between doctors differing in level of experience. DESIGN: Descriptive. METHOD: Indirect limb-length measurement by placing 0.5 cm-thick wooden boards under the foot of the s

  16. Similar estimates of temperature impacts on global wheat yield by three independent methods

    NARCIS (Netherlands)

    Liu, Bing; Asseng, Senthold; Müller, Christoph; Ewert, Frank; Elliott, Joshua; Lobell, David B.; Martre, Pierre; Ruane, Alex C.; Wallach, Daniel; Jones, James W.; Supit, Iwan; Wolf, Joost

    2016-01-01

    The potential impact of global temperature change on global crop yield has recently been assessed with different methods. Here we show that grid-based and point-based simulations and statistical regressions (from historic records), without deliberate adaptation or CO 2 fertilization effects,

  17. Similar estimates of temperature impacts on global wheat yield by three independent methods

    Science.gov (United States)

    Liu, Bing; Asseng, Senthold; Müller, Christoph; Ewert, Frank; Elliott, Joshua; Lobell, David B.; Martre, Pierre; Ruane, Alex C.; Wallach, Daniel; Jones, James W.; Rosenzweig, Cynthia; Aggarwal, Pramod K.; Alderman, Phillip D.; Anothai, Jakarat; Basso, Bruno; Biernath, Christian; Cammarano, Davide; Challinor, Andy; Deryng, Delphine; Sanctis, Giacomo De; Doltra, Jordi; Fereres, Elias; Folberth, Christian; Garcia-Vila, Margarita; Gayler, Sebastian; Hoogenboom, Gerrit; Hunt, Leslie A.; Izaurralde, Roberto C.; Jabloun, Mohamed; Jones, Curtis D.; Kersebaum, Kurt C.; Kimball, Bruce A.; Koehler, Ann-Kristin; Kumar, Soora Naresh; Nendel, Claas; O'Leary, Garry J.; Olesen, Jørgen E.; Ottman, Michael J.; Palosuo, Taru; Prasad, P. V. Vara; Priesack, Eckart; Pugh, Thomas A. M.; Reynolds, Matthew; Rezaei, Ehsan E.; Rötter, Reimund P.; Schmid, Erwin; Semenov, Mikhail A.; Shcherbak, Iurii; Stehfest, Elke; Stöckle, Claudio O.; Stratonovitch, Pierre; Streck, Thilo; Supit, Iwan; Tao, Fulu; Thorburn, Peter; Waha, Katharina; Wall, Gerard W.; Wang, Enli; White, Jeffrey W.; Wolf, Joost; Zhao, Zhigan; Zhu, Yan

    2016-12-01

    The potential impact of global temperature change on global crop yield has recently been assessed with different methods. Here we show that grid-based and point-based simulations and statistical regressions (from historic records), without deliberate adaptation or CO2 fertilization effects, produce similar estimates of temperature impact on wheat yields at global and national scales. With a 1 °C global temperature increase, global wheat yield is projected to decline between 4.1% and 6.4%. Projected relative temperature impacts from different methods were similar for major wheat-producing countries China, India, USA and France, but less so for Russia. Point-based and grid-based simulations, and to some extent the statistical regressions, were consistent in projecting that warmer regions are likely to suffer more yield loss with increasing temperature than cooler regions. By forming a multi-method ensemble, it was possible to quantify `method uncertainty’ in addition to model uncertainty. This significantly improves confidence in estimates of climate impacts on global food security.

  18. Similar estimates of temperature impacts on global wheat yield by three independent methods

    NARCIS (Netherlands)

    Liu, Bing; Asseng, Senthold; Müller, Christoph; Ewert, Frank; Elliott, Joshua; Lobell, David B.; Martre, Pierre; Ruane, Alex C.; Wallach, Daniel; Jones, James W.; Supit, Iwan; Wolf, Joost

    2016-01-01

    The potential impact of global temperature change on global crop yield has recently been assessed with different methods. Here we show that grid-based and point-based simulations and statistical regressions (from historic records), without deliberate adaptation or CO 2 fertilization effects, prod

  19. Survey of blood cultures methods in Italy in 2010

    Directory of Open Access Journals (Sweden)

    Antonio Goglio

    2011-09-01

    Full Text Available Sepsis is a serious clinical condition, associated with high mortality despite advanced modern medical treatment. Traditionally, the detection and identification of bacteria and fungi circulating in the blood-stream is based on blood cultures. A number of factors influence the yield of blood culture, most of them concerning the microbiologist skill and the laboratory organization. In order to collect information about the practices and procedures used for the detection of microrganisms in blood cultures in the italian laboratory (lab, an e-mail with the invitation to participate in the survey was sent to 2000 members of the Italian Association of Clinical Microbiology. Responses were received from 100 lab, located from all over the country (in 18/20 italian regions. The results presented hereby concern specimen collection, culture techniques, rapid identification and susceptibility testing, laboratory organization, relationships with physicians. In summary, most lab use automated systems (96%, the bottles are incubated immediately during public holidays in 72/96 lab (75% and in 49/97 lab at night (50.5%, the lenght of incubation was 5 or 7 days in 93% of the lab, although it is common to extend the incubation period when brucellosis (74 lab, endocarditis (49 lab, systemic mycosis (33 lab is suspected. A wide variety of media are employed for subcultures. All lab process the positive bottles at least once a day, while only in 42 of 81 (51.9% lab the positive blood are processed on holiday. Communication between clinicians and microbiologist include: distribution of specimen collection guidelines (96/100 lab, availability to microbiologist of patients’ clinical situation (77/96 lab, 80.2%, and adding to report the microbiologist’ suggestion (75/98 lab, 76.5%. The results, compared with those collected with a similar questionnaire in 2001, show a greater adherence to guidelines: the number of bottles examined by lab yearly is almost doubled

  20. Determination of nifuroxazide and drotaverine hydrochloride in pharmaceutical preparations by three independent analytical methods.

    Science.gov (United States)

    Metwally, Fadia H; Abdelkawy, Mohammed; Naguib, Ibrahim A

    2006-01-01

    Three new, different, simple, sensitive, and accurate methods were developed for quantitative determination of nifuroxazide (I) and drotaverine hydrochloride (II) in a binary mixture. The first method was spectrophotometry, which allowed determination of I in the presence of II using a zero-order spectrum with an analytically useful maximum at 364.5 nm that obeyed Beer's law over a concentration range of 2-10 microg/mL with mean percentage recovery of 100.08 +/- 0.61. Determination of II in presence of I was obtained by second derivative spectrophotometry at 243.6 nm, which obeyed Beer's law over a concentration range of 2-10 microg/mL with mean recovery of 99.82 +/- 1.46%. The second method was spectrodensitometry, with which both drugs were separated on a silica gel plate using chloroform-acetone-methanol-glacial acetic acid (6 + 3 + 0.9 + 0.1) as the mobile phase and ultraviolet (UV) detection at 365 nm over a concentration range of 0.2-1 microg/band for both drugs, with mean recoveries of 99.99 +/- 0.15 and 100.00 +/- 0.34% for I and II, respectively. The third method was reversed-phase liquid chromatography using acetonitrile-water (40 + 60, v/v; adjusted to pH 2.55 with orthophosphoric acid) as the mobile phase and pentoxifylline as the internal standard at a flow rate of 1 mU/min with UV detection at 285 nm at ambient temperature over a concentration range of 2-10 microg/mL for both drugs, with mean recoveries of 100.24 +/- 1.51 and 100.08 +/- 0.78% for I and II, respectively. The proposed methods were checked using laboratory-prepared mixtures and were successfully applied for the analysis of pharmaceutical formulations containing the above drugs with no interference from other dosage form additives. The validity of the suggested procedures was further assessed by applying the standard addition technique which was found to be satisfactory, and the percentage recoveries obtained were in accordance with those given by the EVA Pharma reference

  1. A maximum likelihood method for studying gene-environment interactions under conditional independence of genotype and exposure.

    Science.gov (United States)

    Cheng, K F

    2006-09-30

    Given the biomedical interest in gene-environment interactions along with the difficulties inherent in gathering genetic data from controls, epidemiologists need methodologies that can increase precision of estimating interactions while minimizing the genotyping of controls. To achieve this purpose, many epidemiologists suggested that one can use case-only design. In this paper, we present a maximum likelihood method for making inference about gene-environment interactions using case-only data. The probability of disease development is described by a logistic risk model. Thus the interactions are model parameters measuring the departure of joint effects of exposure and genotype from multiplicative odds ratios. We extend the typical inference method derived under the assumption of independence between genotype and exposure to that under a more general assumption of conditional independence. Our maximum likelihood method can be applied to analyse both categorical and continuous environmental factors, and generalized to make inference about gene-gene-environment interactions. Moreover, the application of this method can be reduced to simply fitting a multinomial logistic model when we have case-only data. As a consequence, the maximum likelihood estimates of interactions and likelihood ratio tests for hypotheses concerning interactions can be easily computed. The methodology is illustrated through an example based on a study about the joint effects of XRCC1 polymorphisms and smoking on bladder cancer. We also give two simulation studies to show that the proposed method is reliable in finite sample situation.

  2. Simple and Rapid Method for the Determination of Uric Acid-Independent Antioxidant Capacity

    OpenAIRE

    2011-01-01

    Determination of the relative contribution of uric acid level increases to the total measured antioxidative activity could be very useful for testing antioxidative products and their effect on human health. The aim of this report is to present a simple spectrophotometric method that combines the measurement of total antioxidative capacity of a sample by ferric reducing/antioxidative power (FRAP) assay, with the uricase-reaction (specific elimination of uric acid), in order to establish and co...

  3. Consistency and reproducibility of the VMAT plan delivery using three independent validation methods.

    Science.gov (United States)

    Chandraraj, Varatharaj; Stathakis, Sotirios; Manickam, Ravikumar; Esquivel, Carlos; Supe, Sanjay S; Papanikolaou, Nikos

    2010-11-16

    The complexity of VMAT delivery requires new methods and potentially new tools for the commissioning of these systems. It appears that great consideration is needed for quality assurance (QA) of these treatments since there are limited devices that are dedicated to the QA of rotational delivery. In this present study, we have evaluated the consistency and reproducibility of one prostate and one lung VMAT plans for 31 consecutive days using three different approaches: 1) MLC DynaLog files, 2) in vivo measurements using the multiwire ionization chamber DAVID, and 3) using PTWseven29 2D ARRAY with the OCTAVIUS phantom at our Varian Clinac linear accelerator. Overall, the three methods of testing the reproducibility and consistency of the VMAT delivery were in agreement with each other. All methods showed minimal daily deviations that contributed to clinically insignificant dose variations from day to day. Based on our results, we conclude that the VMAT delivery using a Varian 2100CD linear accelerator equipped with 120 MLC is highly reproducible.

  4. Triangulation and the importance of establishing valid methods for food safety culture evaluation.

    Science.gov (United States)

    Jespersen, Lone; Wallace, Carol A

    2017-10-01

    The research evaluates maturity of food safety culture in five multi-national food companies using method triangulation, specifically self-assessment scale, performance documents, and semi-structured interviews. Weaknesses associated with each individual method are known but there are few studies in food safety where a method triangulation approach is used for both data collection and data analysis. Significantly, this research shows that individual results taken in isolation can lead to wrong conclusions, resulting in potentially failing tactics and wasted investments. However, by applying method triangulation and reviewing results from a range of culture measurement tools it is possible to better direct investments and interventions. The findings add to the food safety culture paradigm beyond a single evaluation of food safety culture using generic culture surveys. Copyright © 2017. Published by Elsevier Ltd.

  5. Development of a method for the purification and culture of rodent astrocytes.

    Science.gov (United States)

    Foo, Lynette C; Allen, Nicola J; Bushong, Eric A; Ventura, P Britten; Chung, Won-Suk; Zhou, Lu; Cahoy, John D; Daneman, Richard; Zong, Hui; Ellisman, Mark H; Barres, Ben A

    2011-09-08

    The inability to purify and culture astrocytes has long hindered studies of their function. Whereas astrocyte progenitor cells can be cultured from neonatal brain, culture of mature astrocytes from postnatal brain has not been possible. Here, we report a new method to prospectively purify astrocytes by immunopanning. These astrocytes undergo apoptosis in culture, but vascular cells and HBEGF promote their survival in serum-free culture. We found that some developing astrocytes normally undergo apoptosis in vivo and that the vast majority of astrocytes contact blood vessels, suggesting that astrocytes are matched to blood vessels by competing for vascular-derived trophic factors such as HBEGF. Compared to traditional astrocyte cultures, the gene profiles of the cultured purified postnatal astrocytes much more closely resemble those of in vivo astrocytes. Although these astrocytes strongly promote synapse formation and function, they do not secrete glutamate in response to stimulation.

  6. Comparison of serum fractionation methods by data independent label-free proteomics

    Directory of Open Access Journals (Sweden)

    D. Baiwir

    2015-12-01

    Full Text Available Off-line sample prefractionations applied prior to biomarker discovery proteomics are options to enable more protein identifications and detect low-abundance proteins. This work compared five commercial methods efficiency to raw serum analysis using label-free proteomics. The variability of the protein quantities determined for each process was similar to the unprefractionated serum. A 49% increase in protein identifications and 12.2% of reliable quantification were obtained. A 61 times lower limit of protein quantitation was reached compared to protein concentrations observed in raw serum. The concentrations of detected proteins were confronted to estimated reference values.

  7. A method to determine stratification efficiency of thermal energy storage processes independently from storage heat losses

    DEFF Research Database (Denmark)

    Haller, M.Y.; Yazdanshenas, Eshagh; Andersen, Elsa

    2010-01-01

    A new method for the calculation of a stratification efficiency of thermal energy storages based on the second law of thermodynamics is presented. The biasing influence of heat losses is studied theoretically and experimentally. Theoretically, it does not make a difference if the stratification...... process is in agreement with the first law of thermodynamics. A comparison of the stratification efficiencies obtained from experimental results of charging, standby, and discharging processes gives meaningful insights into the different mixing behaviors of a storage tank that is charged and discharged...

  8. A novel, view-independent method for strain mapping in myocardial elastography: eliminating angle and centroid dependence

    Energy Technology Data Exchange (ETDEWEB)

    Zervantonakis, I K; Fung-Kee-Fung, S D; Lee, W-N; Konofagou, E E [Department of Biomedical Engineering, Columbia University, New York, NY (United States)

    2007-07-21

    Robust indices of regional and global cardiac function are a key factor in detection and treatment of heart disease as well as understanding of the fundamental mechanisms of a healthy heart. Myocardial elastography provides a noninvasive method for imaging and measuring displacement and strain of the myocardium for the early detection of cardiovascular disease. However, two-dimensional in-plane axial and lateral strains measured depend on the sonographic view used. This becomes especially critical in a clinical setting and may induce large variations in the measured strains, potentially leading to false diagnoses. A novel method in myocardial elastography is proposed for eliminating this view dependence by deriving the polar, principal and classified principal strains. The performance of the proposed methodology is assessed by employing 3D finite-element left-ventricular models of a control and an ischemic canine heart. Although polar strains are angle-independent, they are sensitive to the selected reference coordinate system, which requires the definition of a centroid of the left ventricle (LV). In contrast, principal strains derived through eigenvalue decomposition exhibit the inherent characteristic of coordinate system independence, offering view (i.e., angle and centroid)-independent strain measurements. Classified principal strains are obtained by assigning the principal components in the physical ventricular coordinate system. An extensive strain analysis illustrates the improvement in interpretation and visualization of the full-field myocardial deformation by using the classified principal strains, clearly depicting the ischemic and non-ischemic regions. Strain maps, independent of sonographic views and imaging planes, that can be used to accurately detect regional contractile dysfunction are demonstrated.

  9. A novel, view-independent method for strain mapping in myocardial elastography: eliminating angle and centroid dependence

    Science.gov (United States)

    Zervantonakis, I. K.; Fung-Kee-Fung, S. D.; Lee, W.-N.; Konofagou, E. E.

    2007-07-01

    Robust indices of regional and global cardiac function are a key factor in detection and treatment of heart disease as well as understanding of the fundamental mechanisms of a healthy heart. Myocardial elastography provides a noninvasive method for imaging and measuring displacement and strain of the myocardium for the early detection of cardiovascular disease. However, two-dimensional in-plane axial and lateral strains measured depend on the sonographic view used. This becomes especially critical in a clinical setting and may induce large variations in the measured strains, potentially leading to false diagnoses. A novel method in myocardial elastography is proposed for eliminating this view dependence by deriving the polar, principal and classified principal strains. The performance of the proposed methodology is assessed by employing 3D finite-element left-ventricular models of a control and an ischemic canine heart. Although polar strains are angle-independent, they are sensitive to the selected reference coordinate system, which requires the definition of a centroid of the left ventricle (LV). In contrast, principal strains derived through eigenvalue decomposition exhibit the inherent characteristic of coordinate system independence, offering view (i.e., angle and centroid)-independent strain measurements. Classified principal strains are obtained by assigning the principal components in the physical ventricular coordinate system. An extensive strain analysis illustrates the improvement in interpretation and visualization of the full-field myocardial deformation by using the classified principal strains, clearly depicting the ischemic and non-ischemic regions. Strain maps, independent of sonographic views and imaging planes, that can be used to accurately detect regional contractile dysfunction are demonstrated.

  10. Direct Detection and Identification of Enteroviruses from Faeces of Healthy Nigerian Children Using a Cell-Culture Independent RT-Seminested PCR Assay

    Science.gov (United States)

    Adewumi, Moses Olubusuyi; Coker, Bamidele Atinuke; Nudamajo, Felix Yasha; Adeniji, Johnson Adekunle

    2016-01-01

    Recently, a cell-culture independent protocol for detection of enteroviruses from clinical specimen was recommended by the WHO for surveillance alongside the previously established protocols. Here, we investigated whether this new protocol will show the same enterovirus diversity landscape as the established cell-culture dependent protocols. Faecal samples were collected from sixty apparently healthy children in Ibadan, Nigeria. Samples were resuspended in phosphate buffered saline, RNA was extracted, and the VP1 gene was amplified using WHO recommended RT-snPCR protocol. Amplicons were sequenced and sequences subjected to phylogenetic analysis. Fifteen (25%) of the 60 samples yielded the expected band size. Of the 15 amplicons sequenced, 12 were exploitable. The remaining 3 had electropherograms with multiple peaks and were unexploitable. Eleven of the 12 exploitable sequences were identified as Coxsackievirus A1 (CVA1), CVA3, CVA4, CVA8, CVA20, echovirus 32 (E32), enterovirus 71 (EV71), EVB80, and EVC99. Subsequently, the last exploitable sequence was identified as enterobacteriophage baseplate gene by nucleotide BLAST. The results of this study document the first description of molecular sequence data on CVA1, CVA8, and E32 strains present in Nigeria. The result further showed that species A enteroviruses were more commonly detected in the region when cell-culture bias is bypassed. PMID:27087810

  11. Direct Detection and Identification of Enteroviruses from Faeces of Healthy Nigerian Children Using a Cell-Culture Independent RT-Seminested PCR Assay

    Directory of Open Access Journals (Sweden)

    Temitope Oluwasegun Cephas Faleye

    2016-01-01

    Full Text Available Recently, a cell-culture independent protocol for detection of enteroviruses from clinical specimen was recommended by the WHO for surveillance alongside the previously established protocols. Here, we investigated whether this new protocol will show the same enterovirus diversity landscape as the established cell-culture dependent protocols. Faecal samples were collected from sixty apparently healthy children in Ibadan, Nigeria. Samples were resuspended in phosphate buffered saline, RNA was extracted, and the VP1 gene was amplified using WHO recommended RT-snPCR protocol. Amplicons were sequenced and sequences subjected to phylogenetic analysis. Fifteen (25% of the 60 samples yielded the expected band size. Of the 15 amplicons sequenced, 12 were exploitable. The remaining 3 had electropherograms with multiple peaks and were unexploitable. Eleven of the 12 exploitable sequences were identified as Coxsackievirus A1 (CVA1, CVA3, CVA4, CVA8, CVA20, echovirus 32 (E32, enterovirus 71 (EV71, EVB80, and EVC99. Subsequently, the last exploitable sequence was identified as enterobacteriophage baseplate gene by nucleotide BLAST. The results of this study document the first description of molecular sequence data on CVA1, CVA8, and E32 strains present in Nigeria. The result further showed that species A enteroviruses were more commonly detected in the region when cell-culture bias is bypassed.

  12. BAC TG-EMBED: one-step method for high-level, copy-number-dependent, position-independent transgene expression.

    Science.gov (United States)

    Bian, Qian; Belmont, Andrew S

    2010-06-01

    Chromosome position effects combined with transgene silencing of multi-copy plasmid insertions lead to highly variable and usually quite low expression levels of mini-genes integrated into mammalian chromosomes. Together, these effects greatly complicate obtaining high-level expression of therapeutic proteins in mammalian cells or reproducible expression of individual or multiple transgenes. Here, we report a simple, one-step procedure for obtaining high-level, reproducible mini-gene expression in mammalian cells. By inserting mini-genes at different locations within a BAC containing the DHFR housekeeping gene locus, we obtain copy-number-dependent, position-independent expression with chromosomal insertions of one to several hundred BAC copies. These multi-copy DHFR BAC insertions adopt similar large-scale chromatin conformations independent of their chromosome integration site, including insertions within centromeric heterochromatin. Prevention of chromosome position effects, therefore, may be the result of embedding the mini-gene within the BAC-specific large-scale chromatin structure. The expression of reporter mini-genes can be stably maintained during continuous, long-term culture in the presence of drug selection. Finally, we show that this method is extendable to reproducible, high-level expression of multiple mini-genes, providing improved expression of both single and multiple transgenes.

  13. In vitro methods to culture primary human breast epithelial cells.

    Science.gov (United States)

    Raouf, Afshin; Sun, Yu Jia

    2013-01-01

    Current evidence suggests that much like leukemia, breast tumors are maintained by a small subpopulation of tumor cells that have stem cell properties. These cancer stem cells are envisaged to be responsible for tumor formation and relapse. Therefore, knowledge about their nature will provide a platform to develop therapies to eliminate these breast cancer stem cells. This concept highlights the need to understand the mechanisms that regulate the normal functions of the breast stem cells and their immediate progeny as alterations to these same mechanisms can cause these primitive cells to act as cancer stem cells. The study of the primitive cell functions relies on the ability to isolate them from primary sources of breast tissue. This chapter describes processing of discarded tissue from reduction mammoplasty samples as sources of normal primary human breast epithelial cells and describes cell culture systems to grow single-cell suspensions prepared from these reduction samples in vitro.

  14. Application of modified enzyme digestion method in rapid primary culture of human glioma cells

    Directory of Open Access Journals (Sweden)

    Wei XIANG

    2016-06-01

    Full Text Available Objective  To explore the applied value of modified enzyme digestion method in primary culture of human glioma cells. Methods  A traditional enzyme digestion method was modified based on literatures and our work experience. The glioma cells from 32 glioma patients with different grades were primarily cultured by the modified enzyme digestion method. The morphological features of these cells were observed under an inverted phase contrast microscope. The primary cells were purified by differential adhesion during passage. The primary cells were identified by immunofluorescence technique, and the growth curves were drawn by cell proliferation assays (CCK-8 method for investigating the proliferation of the cells cultured in vitro. Results  The primary human glioma cells were successfully cultured and transferred by the new method, with a success rate of 87.5%. The cells cultured successfully in vitro showed good adherent growth, stable morphologies, thus can be passaged. Fluoroimmunoassay showed positive expression of glial fibrillary acidic protein, which confirms the cultured cells were glioma cells. Cell proliferation assays revealed active cell proliferation in vitro, the higher the tumor grade, the higher the proliferative capacity. Conclusion  The modified enzyme digestion method is simpler and more efficient for primary culture of human glioma cells, and the success rate is also higher, thus being able to provide a good guarantee for fundamental research of glioma. DOI: 10.11855/j.issn.0577-7402.2016.06.06

  15. A method for independent component graph analysis of resting-state fMRI

    DEFF Research Database (Denmark)

    de Paula, Demetrius Ribeiro; Ziegler, Erik; Abeyasinghe, Pubuditha M.

    2017-01-01

    -contiguous regions. To date, the spatial patterns of the networks have been analyzed with techniques developed for volumetric data. Objective Here, we detail a graph building technique that allows these ICNs to be analyzed with graph theory. Methods First, ICA was performed at the single-subject level in 15 healthy...... parcellated regions. Third, between-node functional connectivity was established by building edge weights for each networks. Group-level graph analysis was finally performed for each network and compared to the classical network. Results Network graph comparison between the classically constructed network...... for each network. This increased specificity could be relevant for studying pathological brain activity or altered states of consciousness as induced by anesthesia or sleep, where specific networks are known to be altered in different strength....

  16. Culture-independent identification and quantification of Gallibacterium anatis (G. anatis) by real-time quantitative PCR

    DEFF Research Database (Denmark)

    Wang, Chong; Robles, Francisco; Ramirez, Saul

    2016-01-01

    Gallibacterium is a genus within the family Pasteurellaceae characterized by a high level of phenotypic and genetic diversity. No diagnostic method has yet been described, which allows species-specific identification of Gallibacterium anatis. The aim of this study was to develop a real-time quant...

  17. Plant tissue culture independent Agrobacterium tumefaciens mediated In-planta transformation strategy for upland cotton (Gossypium hirsutum

    Directory of Open Access Journals (Sweden)

    Bipinchandra B. Kalbande

    2016-06-01

    Full Text Available A new method of transgenic development called “In-planta” transformation method, where Agrobacterium is used to infect the plantlets but the steps of in vitro regeneration of plants is totally avoided. In this study, we have reported a simple In-planta method for efficient transformation of diploid cotton Gossypium hirsutum cv LRK-516 Anjali using Agrobacterium tumefaciens EHA-105 harbouring recombinant binary vector plasmid pBinAR with Arabidopsis At-NPR1 gene. Four day old plantlets were used for transformation. A vertical cut was made at the junction of cotyledonary leaves, moderately bisecting the shoot tip and exposing meristem cells at apical meristem. This site was infected with Agrobacterium inoculum. The transgenic events obtained were tested positive for the presence of At-NPR1 gene with promoter nptII gene. They are also tested negative for vector backbone integration and Agrobacterium contamination in T0 events. With this method a transformation frequency of 6.89% was reported for the cv LRK-516.

  18. Determination of field scale ammonia emissions for common slurry spreading practice with two independent methods

    Directory of Open Access Journals (Sweden)

    A. Neftel

    2011-09-01

    Full Text Available At a cropland and a grassland site field scale ammonia (NH3 emissions from slurry application were determined simultaneously by two approaches based on (i eddy covariance (EC flux measurements using high temperature Chemical Ionisation Mass Spectrometry (HT-CIMS and on (ii backward Lagrangian Stochastic (bLS dispersion modelling using concentration measurements by three optical open path Fourier Transform Infrared (FTIR systems. Slurry was spread on the fields in sequential tracks over a period of one to two hours. In order to calculate field emissions, measured EC/HT-CIMS fluxes were combined with flux footprint analysis of individual slurry spreading tracks to parameterise the NH3 volatilisation with a bi-exponential time dependence. Accordingly, track-resolved concentration footprints for the FTIR measurements were calculated using bLS. A consistency test with concentrations measured by impingers showed very low systematic deviations for the EC/HT-CIMS results (<8% but larger deviations for the bLS/FTIR results. For both slurry application events, the period during fertilisation and the subsequent two hours contributed by more than 80% to the total field emissions. Averaged over the two measurement methods, the cumulated emissions of the first day amounted to 17 ± 3% loss of applied total ammoniacal nitrogen over the cropland and 16 ± 3% over the grassland field.

  19. A layered searchable encryption scheme with functional components independent of encryption methods.

    Science.gov (United States)

    Luo, Guangchun; Peng, Ningduo; Qin, Ke; Chen, Aiguo

    2014-01-01

    Searchable encryption technique enables the users to securely store and search their documents over the remote semitrusted server, which is especially suitable for protecting sensitive data in the cloud. However, various settings (based on symmetric or asymmetric encryption) and functionalities (ranked keyword query, range query, phrase query, etc.) are often realized by different methods with different searchable structures that are generally not compatible with each other, which limits the scope of application and hinders the functional extensions. We prove that asymmetric searchable structure could be converted to symmetric structure, and functions could be modeled separately apart from the core searchable structure. Based on this observation, we propose a layered searchable encryption (LSE) scheme, which provides compatibility, flexibility, and security for various settings and functionalities. In this scheme, the outputs of the core searchable component based on either symmetric or asymmetric setting are converted to some uniform mappings, which are then transmitted to loosely coupled functional components to further filter the results. In such a way, all functional components could directly support both symmetric and asymmetric settings. Based on LSE, we propose two representative and novel constructions for ranked keyword query (previously only available in symmetric scheme) and range query (previously only available in asymmetric scheme).

  20. Comparative study between the proposed shape independent clustering method and the conventional methods (K-means and the other

    Directory of Open Access Journals (Sweden)

    Kohei Arai

    2013-07-01

    Full Text Available Cluster analysis aims at identifying groups of similar objects and, therefore helps to discover distribution of patterns and interesting correlations in the data sets. In this paper, we propose to provide a consistent partitioning of a dataset which allows identifying any shape of cluster patterns in case of numerical clustering, convex or non-convex. The method is based on layered structure representation that be obtained from measurement distance and angle of numerical data to the centroid data and based on the iterative clustering construction utilizing a nearest neighbor distance between clusters to merge. Encourage result show the effectiveness of the proposed technique.

  1. Estimation of the bottom stress and bottom drag coefficient in a highly asymmetric tidal bay using three independent methods

    Science.gov (United States)

    Xu, Peng; Mao, Xinyan; Jiang, Wensheng

    2017-05-01

    Three independent methods, the dynamical balance (DB) method, the turbulence parameter (TP) method, and the log-layer fit (LF) method, are commonly employed to estimate the bottom stress and bottom drag coefficient in strong tidal systems. However, their results usually differ from each other and the differences are attributed to form drag. Alternatively, some researchers argued that the differences are caused by overestimates in some methods. Aiming to measure the performances of the three independent methods, they were simultaneously constructed in a bay with highly asymmetric tides. The results of the DB and TP methods are consistent with each other in not only the magnitude but also time variation patterns. The consistency of results of the two methods indicates that skin friction is dominant in the bay. The results of the DB and TP methods reveal obvious flood-dominant asymmetry caused by tidal straining. This flood-dominant asymmetry is enhanced during the transition period from spring to neap tide. When the original log-layer fit is employed, the results are much larger than those of the DB and TP methods, and these differences cannot be attributed to form drag since skin friction is dominant in the bay. Moreover, the results of the original log-layer fit reveal an obvious ebb-dominant asymmetry, which is contradictory to the results of the DB and TP methods. Therefore, the results of the original fit are just overestimates and lack physical meaning. By considering the effect of stratification on the mixing length, the modified log-layer fit achieves results with magnitudes that are close to those of the DB and TP methods, indicating that the modified log-layer fit is more representative of the bottom stress than the original log-layer fit in terms of physical meaning. However, the results of the modified log-layer fit still exhibit an ebb-dominant asymmetry in contrast to that of the DB and TP methods, implying that the empirical formula of the mixing

  2. EXPLANTATION OF MESANGIAL CELL HILLOCKS - A METHOD FOR OBTAINING HUMAN MESANGIAL CELLS IN CULTURE

    NARCIS (Netherlands)

    MULLER, EW; KIM, Y; MICHAEL, AF; VERNIER, RL; VANDERHEM, GK; VANDERWOUDE, FJ

    1992-01-01

    A simple method is presented for selective cell culture of human mesangial cells using explanatation of mesangial cell hillocks. Glomeruli which had been incubated with collagenase were explanted on plastic tissue culture flasks. Three to 6 weeks after explantation, a rapidly growing multilayer of e

  3. Perceptions of Organizational Culture of a Multi-Campus Community College District: Mixed Methods in Concert

    Science.gov (United States)

    Kuster Dale, Kimberly

    2012-01-01

    This concurrent, mixed-methods case study analyzed perceptions of current and preferred organizational culture within a rural, multi-campus community college district. This phenomenon was examined by analyzing and comparing data collected by surveying all full-time employees utilizing the Organizational Culture Assessment Instrument (OCAI) and…

  4. Choice of Appropriate Multimedia Technology and Teaching Methods for Different Culture Groups

    Science.gov (United States)

    Taratoukhina, Julia

    2014-01-01

    This paper describes the prerequisites for development in the area of cross-cultural multimedia didactics. This approach is based on research studies of differences between mentalities, ways of working with educational information, culturally-specific teaching methods and teaching techniques that determine differentiated approaches to the choice…

  5. Perceptions of Organizational Culture of a Multi-Campus Community College District: Mixed Methods in Concert

    Science.gov (United States)

    Kuster Dale, Kimberly

    2012-01-01

    This concurrent, mixed-methods case study analyzed perceptions of current and preferred organizational culture within a rural, multi-campus community college district. This phenomenon was examined by analyzing and comparing data collected by surveying all full-time employees utilizing the Organizational Culture Assessment Instrument (OCAI) and…

  6. EXPLANTATION OF MESANGIAL CELL HILLOCKS - A METHOD FOR OBTAINING HUMAN MESANGIAL CELLS IN CULTURE

    NARCIS (Netherlands)

    MULLER, EW; KIM, Y; MICHAEL, AF; VERNIER, RL; VANDERHEM, GK; VANDERWOUDE, FJ

    1992-01-01

    A simple method is presented for selective cell culture of human mesangial cells using explanatation of mesangial cell hillocks. Glomeruli which had been incubated with collagenase were explanted on plastic tissue culture flasks. Three to 6 weeks after explantation, a rapidly growing multilayer of e

  7. Antioxidant Capacity of Cultured Mammalian Cells Estimated by ESR Method

    Directory of Open Access Journals (Sweden)

    Tamar Kartvelishvili

    2004-01-01

    Full Text Available In the present study, the antioxidant capacity against hydrogen peroxide (H2O2, one of the stress-inducing agents, was investigated in two distinct cell lines: L-41 (human epithelial-like cells and HLF (human diploid lung fibroblasts, which differ in tissue origin, life span in culture, proliferate activity, and special enzyme system activity. The cell antioxidant capacity against H2O2 was estimated by the electron spin resonance (ESR spin-trapping technique in the Fenton reaction system via Fe+2 ion action with H2O2 resulting in hydroxyl radical generation. The effects of catalase inhibitors, such as sodium azide and 3-amino-1,2,4-triazole, on the antioxidant capacity of cells were tested. Based on our observation, it can be concluded that the defensive capacity of cells against H2O2 depends on the ratio between catalase/GPx/SOD and H2O2, especially at high-stress situations, and the intracellular balance of these enzymes are more important than the influence of the single component.

  8. Incidence and Trends of Infections with Pathogens Transmitted Commonly Through Food and the Effect of Increasing Use of Culture-Independent Diagnostic Tests on Surveillance - Foodborne Diseases Active Surveillance Network, 10 U.S. Sites, 2013-2016.

    Science.gov (United States)

    Marder, Ellyn P; Cieslak, Paul R; Cronquist, Alicia B; Dunn, John; Lathrop, Sarah; Rabatsky-Ehr, Therese; Ryan, Patricia; Smith, Kirk; Tobin-D'Angelo, Melissa; Vugia, Duc J; Zansky, Shelley; Holt, Kristin G; Wolpert, Beverly J; Lynch, Michael; Tauxe, Robert; Geissler, Aimee L

    2017-04-21

    Foodborne diseases represent a substantial public health concern in the United States. CDC's Foodborne Diseases Active Surveillance Network (FoodNet) monitors cases reported from 10 U.S. sites* of laboratory-diagnosed infections caused by nine enteric pathogens commonly transmitted through food. This report describes preliminary surveillance data for 2016 on the nine pathogens and changes in incidences compared with 2013-2015. In 2016, FoodNet identified 24,029 infections, 5,512 hospitalizations, and 98 deaths caused by these pathogens. The use of culture-independent diagnostic tests (CIDTs) by clinical laboratories to detect enteric pathogens has been steadily increasing since FoodNet began surveying clinical laboratories in 2010 (1). CIDTs complicate the interpretation of FoodNet surveillance data because pathogen detection could be affected by changes in health care provider behaviors or laboratory testing practices (2). Health care providers might be more likely to order CIDTs because these tests are quicker and easier to use than traditional culture methods, a circumstance that could increase pathogen detection (3). Similarly, pathogen detection could also be increasing as clinical laboratories adopt DNA-based syndromic panels, which include pathogens not often included in routine stool culture (4,5). In addition, CIDTs do not yield isolates, which public health officials rely on to distinguish pathogen subtypes, determine antimicrobial resistance, monitor trends, and detect outbreaks. To obtain isolates for infections identified by CIDTs, laboratories must perform reflex culture(†); if clinical laboratories do not, the burden of culturing falls to state public health laboratories, which might not be able to absorb that burden as the adoption of these tests increases (2). Strategies are needed to preserve access to bacterial isolates for further characterization and to determine the effect of changing trends in testing practices on surveillance.

  9. The East-West method: an exposure-independent method to search for large scale anisotropies of cosmic rays

    CERN Document Server

    Bonino, R; Deligny, O; Ghia, P L; Grigat, M; Letessier-Selvon, A; Lyberis, H; Mollerach, S; Over, S; Roulet, E

    2011-01-01

    The measurement of large scale anisotropies in cosmic ray arrival directions at energies above 10^13 eV is performed through the detection of Extensive Air Showers produced by cosmic ray interactions in the atmosphere. The observed anisotropies are small, so accurate measurements require small statistical uncertainties, i.e. large datasets. These can be obtained by employing ground detector arrays with large extensions (from 10^4 to 10^9 m^2) and long operation time (up to 20 years). The control of such arrays is challenging and spurious variations in the counting rate due to instrumental effects (e.g. data taking interruptions or changes in the acceptance) and atmospheric effects (e.g. air temperature and pressure effects on EAS development) are usually present. These modulations must be corrected very precisely before performing standard anisotropy analyses, i.e. harmonic analysis of the counting rate versus local sidereal time. In this paper we discuss an alternative method to measure large scale anisotrop...

  10. Tourismological valorization of intangible cultural heritage of Serbia according to the Hilary du Cros method

    Directory of Open Access Journals (Sweden)

    Željko Bjeljac

    2016-02-01

    Full Text Available The folk artistry of Serbia is rich in spiritual values tied to customs, celebrations, music, song, dance, games, stories and legends, and this kind of cultural heritage is presented through numerous festivals, events and tourist manifestations. In 2012, the network for the safeguarding of intangible cultural heritage was formed, comprised of the National committee for intangible cultural heritage, the Commission for admission into the registry of intangible cultural heritage, a network of coordinators and the Center for intangible cultural heritage of Serbia. These institutions have chosen 6 elements of intangible cultural heritage, out of 27 suggestions: the slava, the Đurđevdan ritual, the kolo dance, singing accompanied by gusle, Slovakian naive painting, the custom of making and lighting farmers’ candles, Pirot carpet weaving, and Zlakusa pottery as elements of cultural heritage which reflect the national and cultural identity of the Serbian people, and Slavic minorities. These elements of intangible cultural heritage have a certain tourism potential and can represent an important factor in the forming of the tourist brand of Serbia. In order to determine the importance of the 27 suggestions of intangible cultural elements of Serbia, an analysis was conducted, using an adapted form of the Hilary du Cros method of tourist valorization.

  11. Detection of periodontopathogenic bacteria in pregnant women by traditional anaerobic culture method and by a commercial molecular genetic method.

    Science.gov (United States)

    Urbán, Edit; Terhes, Gabriella; Radnai, Márta; Gorzó, István; Nagy, Elisabeth

    2010-06-01

    To culture facultative and strict anaerobic bacteria is a well-established method for analyzing subgingival plaque samples. Micro-IDent and micro-IDent Plus (HAIN Lifescience GmbH, Nehren, Germany) tests are two commercially available rapid PCR-based methods for the identification and quantification of putative periodontopathogen bacteria. In this study, we compared these commercial PCR-based hybridization methods with conventional anaerobic culture technique. A total of 36 subgingival plaque samples were collected from periodontal pockets of pregnant women with chronic localized periodontitis. Aliquots of these samples were evaluated with species-specific probes provided by micro-IDent and micro-IDent Plus tests simultaneously, and from the same samples anaerobic and capnophylic bacteria were cultured on selective media. The overall agreement between both methods was excellent for Eubacterium nodatum, Tannerella forsythia and Porphyromonas gingivalis (97-92%), fair for Capnocytophaga sp, Eikenella corrodens, Actinobacillus actinomycetemcomitans, and Prevotella intermedia (91-89%) and poor for Fusobacterium nucleatum, Parvimonas micra (Micromonas micros), and Campylobacter rectus (86-78%). Discrepancies in the results may be explained by inability of culture method to distinguish between closely related taxa (e.i P. intermedia/Prevotella. nigrescens), and problems of keeping periodontopathogen bacteria viable, which is required for successful detection by standard culture method. Nucleic acid-based methods may replace cultivation method as frequently used methods in microbiological diagnosis of progressive periodontitis, thus micro-IDent and micro-IDent Plus tests can be recommended where culture of periodontopathogenic bacteria is not performed in routine microbiology laboratories to analyze subgingival plaque samples.

  12. Characterization of the Bacterial Community Naturally Present on Commercially Grown Basil Leaves: Evaluation of Sample Preparation Prior to Culture-Independent Techniques

    Directory of Open Access Journals (Sweden)

    Siele Ceuppens

    2015-08-01

    Full Text Available Fresh herbs such as basil constitute an important food commodity worldwide. Basil provides considerable culinary and health benefits, but has also been implicated in foodborne illnesses. The naturally occurring bacterial community on basil leaves is currently unknown, so the epiphytic bacterial community was investigated using the culture-independent techniques denaturing gradient gel electrophoresis (DGGE and next-generation sequencing (NGS. Sample preparation had a major influence on the results from DGGE and NGS: Novosphingobium was the dominant genus for three different basil batches obtained by maceration of basil leaves, while washing of the leaves yielded lower numbers but more variable dominant bacterial genera including Klebsiella, Pantoea, Flavobacterium, Sphingobacterium and Pseudomonas. During storage of basil, bacterial growth and shifts in the bacterial community were observed with DGGE and NGS. Spoilage was not associated with specific bacterial groups and presumably caused by physiological tissue deterioration and visual defects, rather than by bacterial growth.

  13. MODIFIED SIMULATION LEARNING METHOD ON KNOWLEDGE AND ATTITUDE OF NURSING STUDEN T’S CULTURAL AWARENESS AT UNIVERSITAS INDONESIA

    Directory of Open Access Journals (Sweden)

    Enie Novieastari

    2012-06-01

    Full Text Available Nursing students should be prepared to be culturally competent nurses. Cultural awareness is believed as the most important elements of cultural competence. The purpose of this article is to describe the effectiveness of Modified Simulation Learning Methods on cultural awareness as one atribute of cultural competence. Aquasi-experimental (control group design was used to explore the relationship between variabels among 98 first year nursing studentattending Basic Nursing Concept I course at Faculty of Nursing Universitas Indonesia. The knowledge of cultural awareness was found statistically different between participants in Modified Simulation Methods group (intervention and participants using the regular method. However, there were no statistical differences in attitude of cultural awareness between intervention and control groups. It could be concluded that Modified Simulation Learning methods is an effective learning method for increasing cultural knowledge of nusing student to be a competent nurse. Further research should be developed in continuing the improvement of cultural competence such as cultural skills.

  14. QUANTIFICATION OF LISTERIA MONOCYTOGENES IN MILK BY MPN-PCR AND MPN-CULTURE METHODS

    Directory of Open Access Journals (Sweden)

    Mahzad Hosseini

    2014-10-01

    Full Text Available The aim of this study was to compare the MPN-PCR (Most Probable Number- Polymerase Chain Reaction and MPN-Culture methods in enumerating of Listeria monocytogenes in milk. In order to compare the accuracy of these methods, 103 cell/ml Listeria monocytogenes and different background bacteria which may be present in raw milk, were inoculated in sterilized milk. After preparing serial dilutions, three replicates per dilution were inoculated in tubes containing listeria enrichment broth. After 48 hours of incubation, for MPN-Culture three inoculated replicates were subcultured on Oxford agar and suspected colonies were confirmed by performing by biochemical tests. For MPN-PCR assay, the DNA extraction was performed from the three inoculated replicates which were already used for MPN-Culture and PCR assay was performed using primers specific for Listeria monocytogenes. The experiment was repeated three times and the average of enumerated bacteria was calculated by each method separately. Statistical analysis using one sample Wilcoxon signed rank test showed that enumeration by MPN-PCR method was more accurate than enumeration by MPN-Culture method. The result of this study showed that MPN-PCR method in comparision with MPN-Culture even in the presence of different background microorganisms is more rapid and reliable. It is concluded that MPN-PCR method facilitates the enumeration of Listeria monocytogenes without excessive work and could be considered as an alternative to MPN-Culture technique.

  15. Establishment of a novel method for primary culture of normal human cervical keratinocytes

    Institute of Scientific and Technical Information of China (English)

    LIU Yu-zhen; L(U) Xiu-ping; PAN Zi-xuan; ZHANG Wei; CHEN Zhao-ri; WANG Hui; LIU Hua

    2013-01-01

    Background Cervical keratinocytes are recovered at a low numbers and frequently associated with contaminating human fibroblasts which rapidly overgrow the epithelial cells in culture with medium supplemented with 10% fetal bovine serum (FBS).However,it is difficult to initiate keratinocyte cultures with serum-free keratinocyte growth medium alone because cell attachment can be poor.Therefore,the culture of these cells is extremely difficult.In this study,we described a modified culture medium and coated culture plastics for growing normal human cervical epithelial cells in vitro.Methods Normal cervical epithelial tissue pieces were obtained and digested with type Ⅰ collagenase to dissociate the cells and a single cell suspension produced.The cells were cultured on plastic tissue culture substrate alone or substrate coated with collagen type Ⅰ from rat tail,with modified keratinocyte serum-free medium (K-SFM) supplemented with 5% FBS.After attachment,the medium were replaced with K-SFM without FBS.The expression of basal keratins of the ectocervical epithelium,K5,K14 and K19 were assayed by immunofiuorescence with monoclonal antibodies to identify the cell purity.Results Our results indicate that cells attached to the culture plastic more quickly in K-SFM supplemented with 5%FBS than in K-SFM alone,as well as to tissue culture plastic coated with collagen type Ⅰ than plastic alone.The modified medium composed of K-SFM and 5% FBS combined with a specific tissue culture plastic coated with collagen type Ⅰ from rat tail was the best method for culture of normal cervical epithelial cells.K5,K14 and K19 were assayed and keratinocyte purity was nearly 100%.Conclusion A novel,simple and effective method can be used to rapidly obtain highly purified keratinocytes from normal human cervical epithelium.

  16. A simple method for the measurement of labelled compound incorporation into cells in culture.

    Science.gov (United States)

    Suplisson, A; Boissel, J P

    1976-02-10

    A simple method for the measurement of labelled compound incorporation into cells in layer culture was developed. Compared to other methods it proves to spare time and to be more sensitive owing to the fact that cells are not detached from the culture vials until the end of the manipulation as these are dissolved in the scintillation medium together with the cells just before counting.

  17. Diagnostic Approaches For Paediatric Tuberculosis By Use Of Different Specimen Types, Culture Methods, And Pcr

    Science.gov (United States)

    Oberhelman, Richard A.; Soto-Castellares, Giselle; Gilman, Robert H.; Caviedes, Luz; Castillo, Maria E.; Kolevic, Lenka; Pino, Trinidad Del; Saito, Mayuko; Salazar-Lindo, Eduardo; Negron, Eduardo; Montenegro, Sonia; Laguna-Torres, V. Alberto; Moore, David A. J.; Evans, Carlton A.

    2010-01-01

    Background The diagnosis of pulmonary tuberculosis (PTB) presents challenges in children, because symptoms are non-specific, specimens are difficult to obtain, and Mycobacterium tuberculosis (MTB) cultures and smears are often negative. The primary objective was to evaluate new diagnostic approaches for TB in children in a resource-poor country. Methods MTB culture by two techniques and a heminested IS 6110 polymerase chain reaction (PCR) assay were performed on specimens from 218 Peruvian children with symptoms suggestive of PTB and 238 healthy controls. Cases were grouped into moderate- and high-risk categories by Stegen-Toledo score. Two specimens of each type (gastric aspirate [GA], nasopharyngeal aspirate [NPA], and stool specimens) from each case were examined by 1) auramine smear microscopy, 2) broth culture by Microscopic-Observation Drug-Susceptibility (MODS) technique, 3) standard culture on Lowenstein Jensen (LJ) medium, and 4) PCR. Specimens from controls included a single NPA and two stools, examined with the same techniques. Subjects were enrolled 2002 to 2007 at two hospitals in Lima, Peru. Controls were enrolled from a low income shantytown community in south Lima. Findings Twenty-two case subjects (10%) had at least one positive MTB culture (from GA in 22 cases, NPA in 12 cases, and stool in 4 cases). Laboratory confirmation of tuberculosis was more frequent in high-risk than moderate-risk cases. MODS was significantly more sensitive than LJ culture, diagnosing 20/22 vs. 13/22 patients (P=0.015), and MTB isolation by MODS was faster than by LJ culture (mean 10 days vs. 25 days, P<0.001). All 22 culture-confirmed cases had at least one culture-positive GA, and the addition of the second GA specimen increased detection of culture-positive cases by 37%. In high-risk children duplicate GA PCR identified half of all culture-positive cases. Interpretation MODS culture increased PTB diagnostic sensitivity and speed compared with LJ culture. Although most

  18. Adrenergic Activation of Melatonin Secretion in Ovine Pineal Explants in Short-Term Superfusion Culture Occurs via Protein Synthesis Independent and Dependent Phenomena

    Directory of Open Access Journals (Sweden)

    Bogdan Lewczuk

    2014-01-01

    Full Text Available The ovine pineal is generally considered as an interesting model for the study on adrenergic regulation of melatonin secretion due to some functional similarities with this gland in the human. The present investigations, performed in the superfusion culture of pineal explants, demonstrated that the norepinephrine-induced elevation of melatonin secretion in ovine pinealocytes comprised of two subsequent periods: a rapid increase phase and a slow increase phase. The first one included the quick rise in release of N-acetylserotonin and melatonin, occurring parallel to elevation of NE concentration in the medium surrounding explants. This rapid increase phase was not affected by inhibition of translation. The second, slow increase phase began after NE level had reached the maximum concentration in the culture medium and lasted about two hours. It was completely abolished by the treatment with translation inhibitors. The obtained results showed for the first time that the regulation of N-acetylserotonin synthesis in pinealocytes of some species like the sheep involves the on/off mechanism, which is completely independent of protein synthesis and works very fast. They provided strong evidence pointing to the need of revision of the current opinion that arylalkylamines N-acetyltransferase activity in pinealocytes is controlled exclusively by changes in enzyme abundance.

  19. Culture-independent approach of the bacterial bioaerosol diversity in the standard swine confinement buildings, and assessment of the seasonal effect.

    Science.gov (United States)

    Nehme, Benjamin; Létourneau, Valérie; Forster, Robert J; Veillette, Marc; Duchaine, Caroline

    2008-03-01

    The bacterial bioaerosol community of eight swine confinement buildings (SCB) was monitored during two visits in the winter, and one during the summer. To our knowledge, culture-independent approaches and molecular biology tools such as biomass quantification and biodiversity analyses have never been applied to swine building bioaerosol analyses. Total DNA of each sample was extracted and analysed by quantitative real-time polymerase chain reaction, denaturing gradient gel electrophoresis (DGGE) and phylogenetic analysis using primers targeting the bacterial 16S rRNA gene. Even though the total bacterial concentration was higher in winter than in summer, the total bacterial concentration for both seasons was 100 to1000 times higher than the total cultural bacteria. The concentration of bioaerosol was influenced by the temperature indoors, which was regulated with an electronic fan system driving warm air and particles outside of the SCB. Comparison of the DGGE profiles showed the same biodiversity in each SCB during both seasons. The phylogenetic analysis revealed a large number of sequences (93.8%) related to Gram-positive anaerobic bacteria, such as Clostridia, and dominated by the Clostridia cluster I (C. disporicum) and the Clostridia cluster XI (C. glycolycum). The bioaerosol diversity also contained also a low proportion of Bacteroidetes and Lactobacillales-Streptococcales sequences. Analyses of the global community and phylotype diversity showed that the main source of bioaerosols could come from the pig manure slurry.

  20. Cardioprotective activity of urocortin by preventing caspase-independent, non-apoptotic death in cultured neonatal rat cardiomyocytes exposed to ischemia

    Energy Technology Data Exchange (ETDEWEB)

    Takatani-Nakase, Tomoka, E-mail: nakase@mukogawa-u.ac.jp [Department of Pharmaceutics, School of Pharmaceutical Sciences, Mukogawa Women' s University, 11-68, Koshien, Nishinomiya, Hyogo 663-8179 (Japan); Takahashi, Koichi, E-mail: koichi@mukogawa-u.ac.jp [Department of Pharmaceutics, School of Pharmaceutical Sciences, Mukogawa Women' s University, 11-68, Koshien, Nishinomiya, Hyogo 663-8179 (Japan)

    2010-11-12

    Research highlights: {yields} Ischemia induces high level of iPLA{sub 2} resulting in caspase-independent myocyte death. {yields} Urocortin causes iPLA{sub 2} down-regulation leading to avoidance of non-apoptotic death. {yields} The survival-promoting effect of urocortin is abrogated by CRH receptor antagonist. -- Abstract: Caspase-independent, non-apoptotic cell death in ischemic heart disease is considered to be one of the important therapeutic targets, however, the detailed mechanisms of this cell death process are not clear. In this study, we investigated the mechanisms of non-apoptotic cell death in cultured neonatal rat cardiomyocytes during ischemia, and the cardioprotection by preventing the mechanisms. We found that ischemia caused elevation of the phospholipase A{sub 2} (iPLA{sub 2}) expression in the myocytes, leading to distinctive non-apoptotic nuclear shrinkage, and cell death. Moreover, we investigated whether the potent cardioprotective corticotropin-releasing hormone (CRH), urocortin, which had been less focused on non-apoptotic cell death, inhibits the ischemic myocyte death. Ischemia-augmented nuclear shrinkage of the myocytes was suppressed by the pretreatment of {approx}10 nM urocortin before the cells were exposed to ischemia. Urocortin could significantly suppress the expression and activity of iPLA{sub 2}, resulting in preventing the ischemia-induced cell death. The survival-promoting effect of urocortin was abrogated by the CRH receptor antagonist astressin. These findings provide the first evidence linking the targets of the urocortin-mediated cardioprotection to the suppression of the caspase-independent, non-apoptotic death in cardiac myocytes exposed to ischemia.

  1. The Trans-Cultural Comparative Literature Method: Using Grammar Translation Techniques Effectively

    Science.gov (United States)

    Sapargul, Destan; Sartor, Valerie

    2010-01-01

    This article describes the Trans-Cultural Comparative Literature Method, an innovative way to use literature to teach advanced English as a Foreign Language (EFL) students. This method originated from the authors' discovery of common themes and points of view as they compared Turkmen and English literary texts. While the method employs activities…

  2. Plant management in natural areas: balancing chemical, mechanical, and cultural control methods

    Science.gov (United States)

    Steven Manning; James. Miller

    2011-01-01

    After determining the best course of action for control of an invasive plant population, it is important to understand the variety of methods available to the integrated pest management professional. A variety of methods are now widely used in managing invasive plants in natural areas, including chemical, mechanical, and cultural control methods. Once the preferred...

  3. Valuation of Haze Management and Prevention Using the Contingent Valuation Method with the Sure Independence Screening Algorithm

    Directory of Open Access Journals (Sweden)

    Guizhi Wang

    2016-03-01

    Full Text Available Haze has caused the deterioration of air quality and has ultimately affected the ecological environment. The contingent valuation method (CVM is an important assessment method that is widely used in ecological economics. The public’s willingness to pay (WTP for haze management and prevention can be analyzed using dichotomous choices. Here, the method is applied to study the valuation of haze management and prevention. Taking Jiangsu Province as an example, the non-market value is calculated by constructing the binary logistic model from questionnaire data, combined with a data-processing method: the sure independence screening (SIS algorithm. The conclusions are as follows: (1 The public’s WTP for haze management and prevention is closely related to the monthly income of families and transport modality; (2 According to the CVM, the non-market value for haze management and prevention in Jiangsu is 7.645 billion yuan; (3 By the average estimate method (AEM, this value is 12.529 billion yuan, about 1.64 times the estimate from the CVM. This is because the AEM ignores the correlation among the influence factors and, therefore, overestimates the valuation of the services; (4 The CVM, combined with the SIS algorithm, does a better job in estimating the valuation of the services.

  4. The application of independent component analysis with projection method to two-task fMRI data over multiple subjects

    Science.gov (United States)

    Li, Rui; Hui, Mingqi; Yao, Li; Chen, Kewei; Long, Zhiying

    2011-03-01

    Spatial Independent component analysis (sICA) has been successfully used to analyze functional magnetic resonance (fMRI) data. However, the application of ICA was limited in multi-task fMRI data due to the potential spatial dependence between task-related components. Long et al. (2009) proposed ICA with linear projection (ICAp) method and demonstrated its capacity to solve the interaction among task-related components in multi-task fMRI data of single subject. However, it's unclear that how to perform ICAp over a group of subjects. In this study, we proposed a group analysis framework on multi-task fMRI data by combining ICAp with the temporal concatenation method reported by Calhoun (2001). The results of real fMRI experiment containing multiple visual processing tasks demonstrated the feasibility and effectiveness of the group ICAp method. Moreover, compared to the GLM method, the group ICAp method is more sensitive to detect the regions specific to each task.

  5. In Vitro Culturing and Live Imaging of Drosophila Egg Chambers: A History and Adaptable Method.

    Science.gov (United States)

    Peters, Nathaniel C; Berg, Celeste A

    2016-01-01

    The development of the Drosophila egg chamber encompasses a myriad of diverse germline and somatic events, and as such, the egg chamber has become a widely used and influential developmental model. Advantages of this system include physical accessibility, genetic tractability, and amenability to microscopy and live culturing, the last of which is the focus of this chapter. To provide adequate context, we summarize the structure of the Drosophila ovary and egg chamber, the morphogenetic events of oogenesis, the history of egg-chamber live culturing, and many of the important discoveries that this culturing has afforded. Subsequently, we discuss various culturing methods that have facilitated analyses of different stages of egg-chamber development and different types of cells within the egg chamber, and we present an optimized protocol for live culturing Drosophila egg chambers.We designed this protocol for culturing late-stage Drosophila egg chambers and live imaging epithelial tube morphogenesis, but with appropriate modifications, it can be used to culture egg chambers of any stage. The protocol employs a liquid-permeable, weighted "blanket" to gently hold egg chambers against the coverslip in a glass-bottomed culture dish so the egg chambers can be imaged on an inverted microscope. This setup provides a more buffered, stable, culturing environment than previously published methods by using a larger volume of culture media, but the setup is also compatible with small volumes. This chapter should aid researchers in their efforts to culture and live-image Drosophila egg chambers, further augmenting the impressive power of this model system.

  6. A novel method for coral explant culture and micropropagation.

    Science.gov (United States)

    Vizel, Maya; Loya, Yossi; Downs, Craig A; Kramarsky-Winter, Esti

    2011-06-01

    We describe here a method for the micropropagation of coral that creates progeny from tissue explants derived from a single polyp or colonial corals. Coral tissue explants of various sizes (0.5-2.5 mm in diameter) were manually microdissected from the solitary coral Fungia granulosa. Explants could be maintained in an undeveloped state or induced to develop into polyps by manipulating environmental parameters such as light and temperature regimes, as well as substrate type. Fully developed polyps were able to be maintained for a long-term in a closed sea water system. Further, we demonstrate that mature explants are also amenable to this technique with the micropropagation of second-generation explants and their development into mature polyps. We thereby experimentally have established coral clonal lines that maintain their ability to differentiate without the need for chemical induction or genetic manipulation. The versatility of this method is also demonstrated through its application to two other coral species, the colonial corals Oculina patigonica and Favia favus.

  7. Culture-independent detection and characterisation of Mycobacterium tuberculosis and M. africanum in sputum samples using shotgun metagenomics on a benchtop sequencer

    Directory of Open Access Journals (Sweden)

    Emma L. Doughty

    2014-09-01

    Full Text Available Tuberculosis remains a major global health problem. Laboratory diagnostic methods that allow effective, early detection of cases are central to management of tuberculosis in the individual patient and in the community. Since the 1880s, laboratory diagnosis of tuberculosis has relied primarily on microscopy and culture. However, microscopy fails to provide species- or lineage-level identification and culture-based workflows for diagnosis of tuberculosis remain complex, expensive, slow, technically demanding and poorly able to handle mixed infections. We therefore explored the potential of shotgun metagenomics, sequencing of DNA from samples without culture or target-specific amplification or capture, to detect and characterise strains from the Mycobacterium tuberculosis complex in smear-positive sputum samples obtained from The Gambia in West Africa. Eight smear- and culture-positive sputum samples were investigated using a differential-lysis protocol followed by a kit-based DNA extraction method, with sequencing performed on a benchtop sequencing instrument, the Illumina MiSeq. The number of sequence reads in each sputum-derived metagenome ranged from 989,442 to 2,818,238. The proportion of reads in each metagenome mapping against the human genome ranged from 20% to 99%. We were able to detect sequences from the M. tuberculosis complex in all eight samples, with coverage of the H37Rv reference genome ranging from 0.002X to 0.7X. By analysing the distribution of large sequence polymorphisms (deletions and the locations of the insertion element IS6110 and single nucleotide polymorphisms (SNPs, we were able to assign seven of eight metagenome-derived genomes to a species and lineage within the M. tuberculosis complex. Two metagenome-derived mycobacterial genomes were assigned to M. africanum, a species largely confined to West Africa; the others that could be assigned belonged to lineages T, H or LAM within the clade of “modern” M. tuberculosis

  8. Isolation and Identification of Listeria monocytogenes in Processed Meat by a Combined Cultural-molecular Method

    Directory of Open Access Journals (Sweden)

    Angela Ingianni

    2007-01-01

    Full Text Available The isolation and identification of Listeria monocytogenes in processed meat samples by a combined cultural-molecular method is described. It allows the identification of Listeria strains by means of a hybridization technique with a specific DNA probe directed to the listerial internalin gene. The specificity of this method was found to be 100% and sensitivity was as low as 1 CFU/2.5 g of food sample. A total of 278 meat samples were tested in comparison with PCR and conventional cultural assays. A total of 42 (15.4% L. monocytogenes were detected. PCR analysis gave 3 false negative results and culture failed to detect the Listeria in 5 cases. With this cultural-molecular method the identification and quantitative detection of L. monocytogenes were achieved within 36 hours and no false positive or negative tests were obtained, thus fitting most food industry requirements.

  9. Comparison of Fluorescence Microscopy and Different Growth Media Culture Methods for Acanthamoeba Keratitis Diagnosis.

    Science.gov (United States)

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

    2015-08-01

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

  10. Evaluation of three rapid diagnostic methods for direct identification of microorganisms in positive blood cultures.

    Science.gov (United States)

    Martinez, Raquel M; Bauerle, Elizabeth R; Fang, Ferric C; Butler-Wu, Susan M

    2014-07-01

    The identification of organisms from positive blood cultures generally takes several days. However, recently developed rapid diagnostic methods offer the potential for organism identification within only a few hours of blood culture positivity. In this study, we evaluated the performance of three commercial methods to rapidly identify organisms directly from positive blood cultures: QuickFISH (AdvanDx, Wolburn, MA), Verigene Gram-Positive Blood Culture (BC-GP; Nanosphere, Northbrook, IL), and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) with Sepsityper processing (Bruker Daltonics, Billerica, MA). A total of 159 blood cultures (VersaTREK Trek Diagnostic Systems, Cleveland, OH) positive for Gram-positive and Gram-negative bacteria as well as yeast were analyzed with QuickFISH and MALDI-TOF MS. In all, 102 blood cultures were analyzed using the BC-GP assay. For monomicrobial cultures, we observed 98.0% concordance with routine methods for both QuickFISH (143/146) and the BC-GP assay (93/95). MALDI-TOF MS demonstrated 80.1% (117/146) and 87.7% (128/146) concordance with routine methods to the genus and species levels, respectively. None of the methods tested were capable of consistently identifying polymicrobial cultures in their entirety or reliably differentiating Streptococcus pneumoniae from viridans streptococci. Nevertheless, the methods evaluated in this study are convenient and accurate for the most commonly encountered pathogens and have the potential to dramatically reduce turnaround time for the provision of results to the treating physician.

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

    Science.gov (United States)

    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.

  12. Cultural cognition in the thinking-aloud method for usability evaluation

    DEFF Research Database (Denmark)

    Clemmensen, Torkil; Hertzum, Morten; Hornbæk, Kasper

    2008-01-01

    We discuss the impact of cultural differences on usability evaluations that are based on the thinking-aloud method (TA). The term ‘cultural differences' helps distinguish differences in the perception and thinking of Westerners (people from Western Europe and US citizens with European origins......) and Easterners (people from China and the countries heavily influenced by its culture). We illustrate the impact of cultural cognition on four central elements of TA: (1) instructions and tasks, (2) the user's verbalizations, (3) the evaluator's reading of the user, and (4) the overall relationship between user...... and evaluator. In conclusion, we point to the importance of matching the task presentation to users' cultural background, the different effects of thinking aloud on task performance between Easterners and Westerners, the differences in nonverbal behaviour that affect usability problem detection, and, finally...

  13. USDA FSIS and FDA BAM culture methods BBL CHROMagar Salmonella prepared plated and Difco dehydrated culture media.

    Science.gov (United States)

    Ritter, Vicki; Dick, Nancy

    2009-01-01

    BBL and Difco CHROMagar Salmonella (CS) was evaluated internally and externally for the recovery of Salmonella in raw chicken, raw ground beef, raw fish, lettuce, and shell eggs. The raw chicken and ground beef were processed according to the U.S. Department of Agriculture, Food Safety and Inspection Service reference methods. The raw fish, lettuce, and shell eggs were processed according to the U.S. Food and Drug Administration, Bacteriological Analytical Manual procedures. Only raw chicken was found to be naturally contaminated with Salmonella; all other matrixes were seeded with an appropriate dilution of organism to achieve fractionally positive results. Salmonella strains were permitted to equilibrate with the culture-negative matrixes for 48 h at 4 degrees C. Twenty 25 g samples of each food matrix plus 5 uninoculated samples were processed. CS prepared plates (CS PPM) and laboratory prepared plates from dehydrated culture media (CS DCM) were evaluated with the reference method media. A total of 16 positive cultures were obtained from the raw chicken samples, 17 in the raw ground beef, 18 in the raw fish and lettuce, and 11 in the shell eggs. A Chi-square analysis was performed on each of the food matrixes. BBL CS produced comparable results with the reference methods on all matrixes, resulting in a method agreement of 100% based on the Chi-square results. In testing known isolates the sensitivity and specificity was determined to be 94%. Specificity improved to 98% when tetrathionate broth enrichment was used. A negative- and false-positive rate of 6% was found with known isolates. No false negatives were found in testing the food matrixes. The performance of the CS prepared plate and laboratory prepared plate was identical.

  14. Pseudothrombocytopenia: a report of a new method to count platelets in a patient with EDTA- and temperature-independent antibodies of the IgM type.

    Science.gov (United States)

    van der Meer, W; Allebes, W; Simon, A; van Berkel, Y; de Keijzer, M H

    2002-10-01

    Pseudothrombocytopenia is usually associated with blood specimens anticoagulated with ethylenediamine tetraacetic acid (EDTA) or other anticoagulants. It may be caused by temperature-independent, EDTA-dependent antibodies of the immunoglobulin-M (IgM) type. Here a patient with EDTA-independent and temperature-independent pseudothrombocytopenia mediated by IgM or IgM-containing immune complex is reported, and a reliable method is described for a proper counting of platelets in such cases.

  15. Comparison of two methods used to culture and purify rat retinal Müller cells.

    Science.gov (United States)

    Song, Wei-Tao; Zhang, Xue-Yong; Xiong, Si-Qi; Wen, Dan; Jiang, Jian; Xia, Xiao-Bo

    2013-01-01

    To study two methods for culturing and purifying Sprague-Dawley (SD) rat retinal Müller cells and determine which one is better. The passage culture method of Müller cells was respectively carried out by complete pancreatic enzyme digestion method and repeated incomplete pancreatic enzyme digestion method. After culturing retinal cells for one month through these two methods, fluorescence-activated cell sorter (FACS), RT-PCR, and immunohistochemistry technology were performed to examine the enrichment and purity of Müller glial cells, and carried out two-sample approximate t test using SSPS 13.0 to further compare the Müller cell positive rate in both methods. The statistical results showed that the purity of Müller cells was 83.2%±5.16% in group A, and the purity was 98.5%±1.08% in group B. The two-sample approximate t test analysis demonstrated that the difference between group A and group B was statistically significant (t=-9.178, Pcells cultured by the complete pancreatic enzyme digestion method (group A) and the repeated incomplete pancreatic enzyme digestion method (group B). Compared with the complete pancreatic enzyme digestion method, this novel method was more efficient and a higher purity of Müller cells could be obtained using this approach.

  16. EEG/fMRI fusion based on independent component analysis: integration of data-driven and model-driven methods.

    Science.gov (United States)

    Lei, Xu; Valdes-Sosa, Pedro A; Yao, Dezhong

    2012-09-01

    Simultaneous electroencephalography (EEG) and functional magnetic resonance imaging (fMRI) provide complementary noninvasive information of brain activity, and EEG/fMRI fusion can achieve higher spatiotemporal resolution than each modality separately. This focuses on independent component analysis (ICA)-based EEG/fMRI fusion. In order to appreciate the issues, we first describe the potential and limitations of the developed fusion approaches: fMRI-constrained EEG imaging, EEG-informed fMRI analysis, and symmetric fusion. We then outline some newly developed hybrid fusion techniques using ICA and the combination of data-/model-driven methods, with special mention of the spatiotemporal EEG/fMRI fusion (STEFF). Finally, we discuss the current trend in methodological development and the existing limitations for extrapolating neural dynamics.

  17. Determination of absorption cross-section of Si nanocrystals by two independent methods based on either absorption or luminescence

    Energy Technology Data Exchange (ETDEWEB)

    Valenta, J., E-mail: jan.valenta@mff.cuni.cz; Greben, M. [Department of Chemical Physics and Optics, Faculty of Mathematics and Physics, Charles University, Ke Karlovu 3, 121 16 Prague 2 (Czech Republic); Remeš, Z. [Institute of Physics, Academy of Sciences of the Czech Republic, Cukrovarnická 10, Prague 6 (Czech Republic); Gutsch, S.; Hiller, D.; Zacharias, M. [Faculty of Engineering, IMTEK, Albert-Ludwigs-University Freiburg, Georges-Köhler-Allee 103, 79110 Freiburg (Germany)

    2016-01-11

    Absorption cross-section (ACS) of silicon nanocrystals (SiNCs) is determined via two completely independent approaches: (i) Excitation-intensity-dependent photoluminescence (PL) kinetics under modulated (long square pulses) pumping and (ii) absorbance measured by the photothermal deflection spectroscopy combined with morphology information obtained by the high-resolution transmission electron microscopy. This unique comparison reveals consistent ACS values around 10{sup −15} cm{sup 2} for violet excitation of SiNCs of about 3–5 nm in diameter and this value is comparable to most of direct band-gap semiconductor nanocrystals; however, it decreases steeply towards longer wavelengths. Moreover, we analyze the PL-modulation technique in detail and propose an improved experimental procedure which enables simpler implementation of this method to determine ACS of various (nano)materials in both solid and liquid states.

  18. A fluorescence anisotropy method for measuring protein concentration in complex cell culture media.

    Science.gov (United States)

    Groza, Radu Constantin; Calvet, Amandine; Ryder, Alan G

    2014-04-22

    The rapid, quantitative analysis of the complex cell culture media used in biopharmaceutical manufacturing is of critical importance. Requirements for cell culture media composition profiling, or changes in specific analyte concentrations (e.g. amino acids in the media or product protein in the bioprocess broth) often necessitate the use of complicated analytical methods and extensive sample handling. Rapid spectroscopic methods like multi-dimensional fluorescence (MDF) spectroscopy have been successfully applied for the routine determination of compositional changes in cell culture media and bioprocess broths. Quantifying macromolecules in cell culture media is a specific challenge as there is a need to implement measurements rapidly on the prepared media. However, the use of standard fluorescence spectroscopy is complicated by the emission overlap from many media components. Here, we demonstrate how combining anisotropy measurements with standard total synchronous fluorescence spectroscopy (TSFS) provides a rapid, accurate quantitation method for cell culture media. Anisotropy provides emission resolution between large and small fluorophores while TSFS provides a robust measurement space. Model cell culture media was prepared using yeastolate (2.5 mg mL(-1)) spiked with bovine serum albumin (0 to 5 mg mL(-1)). Using this method, protein emission is clearly discriminated from background yeastolate emission, allowing for accurate bovine serum albumin (BSA) quantification over a 0.1 to 4.0 mg mL(-1) range with a limit of detection (LOD) of 13.8 μg mL(-1). Copyright © 2014. Published by Elsevier B.V.

  19. Cultural continuity, traditional Indigenous language, and diabetes in Alberta First Nations: a mixed methods study.

    Science.gov (United States)

    Oster, Richard T; Grier, Angela; Lightning, Rick; Mayan, Maria J; Toth, Ellen L

    2014-10-19

    We used an exploratory sequential mixed methods approach to study the association between cultural continuity, self-determination, and diabetes prevalence in First Nations in Alberta, Canada. We conducted a qualitative description where we interviewed 10 Cree and Blackfoot leaders (members of Chief and Council) from across the province to understand cultural continuity, self-determination, and their relationship to health and diabetes, in the Alberta First Nations context. Based on the qualitative findings, we then conducted a cross-sectional analysis using provincial administrative data and publically available data for 31 First Nations communities to quantitatively examine any relationship between cultural continuity and diabetes prevalence. Cultural continuity, or "being who we are", is foundational to health in successful First Nations. Self-determination, or "being a self-sufficient Nation", stems from cultural continuity and is seriously compromised in today's Alberta Cree and Blackfoot Nations. Unfortunately, First Nations are in a continuous struggle with government policy. The intergenerational effects of colonization continue to impact the culture, which undermines the sense of self-determination, and contributes to diabetes and ill health. Crude diabetes prevalence varied dramatically among First Nations with values as low as 1.2% and as high as 18.3%. Those First Nations that appeared to have more cultural continuity (measured by traditional Indigenous language knowledge) had significantly lower diabetes prevalence after adjustment for socio-economic factors (p =0.007). First Nations that have been better able to preserve their culture may be relatively protected from diabetes.

  20. Standardization of the method for utilization of paneer whey in cultured buttermilk.

    Science.gov (United States)

    Ghanshyambhai, Maheta Riddhiben; Balakrishnan, Smitha; Aparnathi, K D

    2015-05-01

    Whey is a liquid by-product obtained during manufacture of coagulated milk products like paneer, cheese etc. Its disposal as waste leads to heavy load in dairy effluent and loss of valuable milk solids. For efficient and economic utilization of whey, a method was standardized for the preparation of cultured butter milk using paneer whey as one of the ingredient. It involved fermentation of paneer whey and double toned milk separately using starter culture containing Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus. The paneeer whey can be successfully incorporated up to 50 % by following the standardized method. The proximate chemical composition of cultured buttermilk was 8.31 % total solids, 2.26 % protein, 1.12 % fat, 4.42 % lactose and 0.56 % ash. The cultured buttermilk has acceptable sensory qualities and shelf life of 5 days under refrigerated condition.

  1. A method for establishing human primary gastric epithelial cell culture from fresh surgical gastric tissues.

    Science.gov (United States)

    Aziz, Faisal; Yang, Xuesong; Wen, Qingping; Yan, Qiu

    2015-08-01

    At present, biopsy specimens, cancer cell lines and tissues obtained by gastric surgery are used in the study and analysis of gastric cancer, including the molecular mechanisms and proteomics. However, fibroblasts and other tissue components may interfere with these techniques. Therefore, the present study aimed to develop a procedure for the isolation of viable human gastric epithelial cells from gastric surgical tissues. A method was developed to culture human gastric epithelial cells using fresh, surgically excised tissues and was evaluated using immunocytochemistry, periodic acid-Schiff (PAS) staining and cell viability assays. Low cell growth was observed surrounding the gastric tissue on the seventh day of tissue explant culture. Cell growth subsequently increased, and at 12 days post-explant a high number of pure epithelial cells were detected. The gastric cancer cells exhibited rapid growth with a doubling time of 13-52 h, as compared to normal cells, which had a doubling time of 20-53 h. Immunocytochemical analyses of primary gastric cells revealed positive staining for cytokeratin 18 and 19, which indicated that the culture was comprised of pure epithelial cells and contained no fibroblasts. Furthermore, PAS staining demonstrated that the cultured gastric cells produced neutral mucin. Granulin and carbohydrate antigen 724 staining confirmed the purity of gastric cancer and normal cells in culture. This method of cell culture indicated that the gastric cells in primary culture consisted of mucin-secreting gastric epithelial cells, which may be useful for the study of gastric infection with Helicobacter pylori and gastric cancer.

  2. A simplified micropatterning method for straight-line neurite extension of cultured hippocampal neurons.

    Science.gov (United States)

    Suzuki, Ikuro; Nakamura, Kosuke; Odawara, Aoi; Alhebshi, Amani; Gotoh, Masao

    2013-01-01

    We report a simplified micropatterning method for the straight-line extension of the neurites of cultured neurons. We prepared a poly-D-lysine (PDL)-patterned surface using a polydimethylsiloxane microfluidic stamp. Hippocampal neurons were cultured on the PDL-bound substrate with the stamp removed, allowing for conventional cell seeding and detailed optical observation without fluorescent label. Cultured neurons elongated neurites along straight lines at the single-cell level and displayed spontaneous firing as detected by time-lapse imaging and Ca(2+) imaging.

  3. Conversion of primordial germ cells to pluripotent stem cells: methods for cell tracking and culture conditions.

    Science.gov (United States)

    Nagamatsu, Go; Suda, Toshio

    2013-01-01

    Primordial germ cells (PGCs) are unipotent cells committed to germ lineage: PGCs can only differentiate into gametes in vivo. However, upon fertilization, germ cells acquire the capacity to differentiate into all cell types in the body, including germ cells. Therefore, germ cells are thought to have the potential for pluripotency. PGCs can convert to pluripotent stem cells in vitro when cultured under specific conditions that include bFGF, LIF, and the membrane-bound form of SCF (mSCF). Here, the culture conditions which efficiently convert PGCs to pluripotent embryonic germ (EG) cells are described, as well as methods used for identifying pluripotent candidate cells during culture.

  4. Elaboration of Method of Long-Term Culturing and Selection of Enzyme Producers

    Directory of Open Access Journals (Sweden)

    Blieva Raushan

    2014-06-01

    Full Text Available On the basis of the conducted researches on pectin lyase and proteolytic enzyme biosynthesis by Penicillium and Asperaillus micromycetes we have developed efficient methods for their cultivation and selection. We theoretically substantiated and experimentally confirmed an advantage of growing micromycetes in a new filament-spongy immobilized growth structure on the substrate relative to the traditional method of deep cultivation of free cells in the form of pellets. When comparing a traditional with our innovative method of cultivation, many advantages of the latter are revealed, above all being the possibility of the formation of new highly selective cultures in the long process of their growth with modified culturally - morphological properties.

  5. FastCloning: a highly simplified, purification-free, sequence- and ligation-independent PCR cloning method

    Directory of Open Access Journals (Sweden)

    Lu Jia

    2011-10-01

    Full Text Available Abstract Background Although a variety of methods and expensive kits are available, molecular cloning can be a time-consuming and frustrating process. Results Here we report a highly simplified, reliable, and efficient PCR-based cloning technique to insert any DNA fragment into a plasmid vector or into a gene (cDNA in a vector at any desired position. With this method, the vector and insert are PCR amplified separately, with only 18 cycles, using a high fidelity DNA polymerase. The amplified insert has the ends with ~16-base overlapping with the ends of the amplified vector. After DpnI digestion of the mixture of the amplified vector and insert to eliminate the DNA templates used in PCR reactions, the mixture is directly transformed into competent E. coli cells to obtain the desired clones. This technique has many advantages over other cloning methods. First, it does not need gel purification of the PCR product or linearized vector. Second, there is no need of any cloning kit or specialized enzyme for cloning. Furthermore, with reduced number of PCR cycles, it also decreases the chance of random mutations. In addition, this method is highly effective and reproducible. Finally, since this cloning method is also sequence independent, we demonstrated that it can be used for chimera construction, insertion, and multiple mutations spanning a stretch of DNA up to 120 bp. Conclusion Our FastCloning technique provides a very simple, effective, reliable, and versatile tool for molecular cloning, chimera construction, insertion of any DNA sequences of interest and also for multiple mutations in a short stretch of a cDNA.

  6. Production of Pastırma with Different Curing Methods and Using Starter Culture

    OpenAIRE

    AKSU, Muhammet İrfan; Kaya, Mükerrem

    2002-01-01

    The effects of starter culture (Staphylococcus carnosus + Lactobacillus pentosus) on some physical, chemical and microbiological properties of pastırma produced with brine and dry curing methods were investigated. The curing method significantly affected the contents of salt, nitrite, nitrite/nitrate rate at the level of P

  7. A Metabotropic-Like Flux-Independent NMDA Receptor Regulates Ca2+ Exit from Endoplasmic Reticulum and Mitochondrial Membrane Potential in Cultured Astrocytes

    Science.gov (United States)

    Montes de Oca Balderas, Pavel; Aguilera, Penélope

    2015-01-01

    Astrocytes were long thought to be only structural cells in the CNS; however, their functional properties support their role in information processing and cognition. The ionotropic glutamate N-methyl D-aspartate (NMDA) receptor (NMDAR) is critical for CNS functions, but its expression and function in astrocytes is still a matter of research and debate. Here, we report immunofluorescence (IF) labeling in rat cultured cortical astrocytes (rCCA) of all NMDAR subunits, with phenotypes suggesting their intracellular transport, and their mRNA were detected by qRT-PCR. IF and Western Blot revealed GluN1 full-length synthesis, subunit critical for NMDAR assembly and transport, and its plasma membrane localization. Functionally, we found an iCa2+ rise after NMDA treatment in Fluo-4-AM labeled rCCA, an effect blocked by the NMDAR competitive inhibitors D(-)-2-amino-5-phosphonopentanoic acid (APV) and Kynurenic acid (KYNA) and dependent upon GluN1 expression as evidenced by siRNA knock down. Surprisingly, the iCa2+ rise was not blocked by MK-801, an NMDAR channel blocker, or by extracellular Ca2+ depletion, indicating flux-independent NMDAR function. In contrast, the IP3 receptor (IP3R) inhibitor XestosponginC did block this response, whereas a Ryanodine Receptor inhibitor did so only partially. Furthermore, tyrosine kinase inhibition with genistein enhanced the NMDA elicited iCa2+ rise to levels comparable to those reached by the gliotransmitter ATP, but with different population dynamics. Finally, NMDA depleted the rCCA mitochondrial membrane potential (mΔψ) measured with JC-1. Our results demonstrate that rCCA express NMDAR subunits which assemble into functional receptors that mediate a metabotropic-like, non-canonical, flux-independent iCa2+ increase. PMID:25954808

  8. Las universidades libres y populares en Portugal y el problema de la cultura popular - The independent and popular universities in Portugal and the problem of popular culture

    Directory of Open Access Journals (Sweden)

    Rogério Fernandes

    2011-03-01

    Full Text Available Resumo O movimento das Universidade Livres e Populares em Portugal não está devidamente estudado, apesar de sua importância para a história social e a história da educação. Existem, é certo, alguns trabalhos monográficos parciais, em forma de livro impresso ou fotocopiado, e atigos dispersos por periódicos ou revistas, trazendo informações mais ou menos valiosas. Pretendemos, no entanto, de uma análise de conjunto com a necessária profundidade, que permita avaliar a evolução de suas unidades, a ação desenvolvida por elas e o significado do papel que desempenharam em relação à cultura popular. O estudo aqui paresentado objetiva caracterizar uma das universidade populares e livres que se criaram em Portugal, no final do século XIX e desenhar o trajeto cultural e político de cada uma delas, privilegiando as mais importantes. Com esse marco, busca-se lançar um olhar crítico sobre as principais Universidades do Norte e centro do país, localizadas no Porto, em Lisboa, Setúbal e Coimbra, intentando caracterizar-lhes a orientação política-ideológica, os objetivos pedagógicos e culturais visados, os meios que recorreram e a identidade política de seus promotores. Palavras-chave: universidades livres; universidades populares; história social; historia da educação; Portugal.   THE INDEPENDENT AND POPULAR UNIVERSITIES IN PORTUGAL AND THE PROBLEM OF POPULAR CULTURE Abstract The movement of Independent and Popular Universities in Portugal is not enough studied if we consider how important it is for Social History and for the History of Education. We have not yet a deep global analysis which enables to consider the evolution of theirs units, the actions that have started and the meaning of their role compared to popular culture. In the following study, we will try to characterize each Popular or Independent University create in Portugal from the late XIXth century, and give the cultural and political ways of each university

  9. Bacterial Ecology of Fermented Cucumber Rising pH Spoilage as Determined by Non-Culture Based Methods

    Science.gov (United States)

    Medina, Eduardo; Pérez-Díaz, Ilenys M.; Breidt, Fred; Hayes, Janet; Franco, Wendy; Butz, Natasha; Azcarate-Peril, María Andrea

    2016-01-01

    Fermented cucumber spoilage (FCS) characterized by rising pH and the appearance of manure and cheese like aromas is a challenge of significant economical impact for the pickling industry. Previous culture based studies identified the yeasts Pichia manshurica and Issatchenkia occidentalis, four gram positive bacteria, Lactobacillus buchneri, Lactobacillus parrafaraginis, Clostridium sp. and Propionibacterium and one gram-negative genus, Pectinatus as relevant in various stages of FCS given their ability to metabolize lactic acid. It was the objective of this study to augment the current knowledge of FCS using culture independent methods to microbiologically characterize commercial spoilage samples. Ion Torrent data and 16S rRNA cloning library analyses of samples collected from commercial fermentation tanks confirmed the presence of L. rapi and L. buchneri and revealed the presence of additional species involved in the development of FCS such as Lactobacillus namurensis, Lactobacillus acetotolerans, Lactobacillus panis, Acetobacter peroxydans, Acetobacter aceti, and Acetobacter pasteurianus at pH below 3.4. The culture independent analyses also revealed the presence of species of Veillonella and Dialister in spoilage samples with pH above 4.0 and confirmed the presence of Pectinatus spp. during lactic acid degradation at the higher pH. Acetobacter spp. were successfully isolated from commercial samples collected from tanks subjected to air purging by plating on Mannitol Yeast Peptone agar. In contrast, Lactobacillus spp. were primarily identified in samples of FCS collected from tanks not subjected to air purging for more than 4 months. Thus, it is speculated that oxygen availability may be a determining factor in the initiation of spoilage and the leading microbiota. Practical Application Understanding of the underlying microbiology and biochemistry driving FCS is essential to enhancing the sodium chloride (NaCl)-free cucumber fermentation technology and in

  10. Detection of campylobacter species: a comparison of culture and polymerase chain reaction based methods

    Science.gov (United States)

    Kulkarni, S P; Lever, S; Logan, J M J; Lawson, A J; Stanley, J; Shafi, M S

    2002-01-01

    Aims: To investigate the optimal method for the detection of campylobacters from stool samples by comparing selective culture with membrane filtration and the polymerase chain reaction (PCR). Methods: Three hundred and forty three stool samples were investigated by each of the three methods mentioned above. Selective culture was performed with charcoal cefoperazone desoxycholate agar plates. Membrane filtration was performed using cellulose triacetate membranes with 0.45 μm pores placed on blood agar plates. Enteropathogenic campylobacters were detected using a PCR identification algorithm, consisting of screening PCRs and species identification using a PCR enzyme linked immunosorbent assay (PCR-ELISA), both based on the 16S rRNA gene. Results: Of the 343 samples tested, 23 were positive by one or more method. Of these, 17 were positive by selective culture, 12 by membrane filtration, and 20 by the PCR identification algorithm. A total of 18 of 23 positives were identified as C jejuni and/or C coli by the PCR identification algorithm, compared with 14 identified to the genus level by selective culture, and 10 by membrane filtration. Among the remaining five positive samples, one C hyointestinalis was detected only by the PCR identification algorithm; one C upsaliensis was detected only by the PCR identification algorithm; one Campylobacter sp was detected by membrane filtration and selective culture and later identified as C concisus; one Campylobacter sp was detected by membrane filtration alone and later identified as Arcobacter sp; and one Campylobacter sp detected only by selective culture was lost to study and therefore not speciated. There was no significant difference between detection by selective culture and the other two methods. However, detection by PCR was significantly better than by membrane filtration (0.05 > p > 0.02). Conclusion: The PCR identification algorithm can detect and identify Campylobacter spp to the species level and the result is

  11. Reconstruction method as an independent risk factor for the postoperative decrease in hemoglobin in stage I gastric cancer.

    Science.gov (United States)

    Imamura, Taisuke; Komatsu, Shuhei; Ichikawa, Daisuke; Kosuga, Toshiyuki; Okamoto, Kazuma; Konishi, Hirotaka; Shiozaki, Atsushi; Fujiwara, Hitoshi; Otsuji, Eigo

    2016-05-01

    No study has compared the incidence of postoperative anemia between two reconstruction methods, Billroth-I (B-I) and Roux-en-Y (R-Y) reconstructions, after distal gastrectomy for gastric cancer (GC). In this study, we wished to examine the postoperative decrease in hemoglobin (Hb) as an indicator of iron-deficiency anemia. We investigated a total of 119 consecutive patients who underwent distal gastrectomy with B-I or R-Y reconstruction for Stage I GC between 2006 and 2012. We retrospectively assessed the clinical data, including Hb results, of the first 2 years after surgery. Compared with B-I reconstruction, R-Y reconstruction was performed more frequently in older patients (P = 0.017), and it was associated with a longer surgical duration (P decrease in Hb for the first 2 years after surgery. Univariate and multivariate analyses identified that R-Y reconstruction was the only risk factor (P = 0.0487; odds ratio = 2.755; 95% confidence interval = 1.01-7.91) for a decrease in Hb, independent of age, tumor location, postoperative complications, and other factors. In addition, an age ≥ 75 was identified as an independent risk factor for a decrease in Hb, particularly for patients underwent R-Y reconstruction (P = 0.033; odds ratio = 6.99; 95% confidence interval = 1.15-68.3) according to the multivariate analysis. Billroth-I reconstruction might be preferable for the purpose of preventing a decrease in Hb in stage I GC patients, particularly in older patients. © 2015 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.

  12. Model Insensitive and Calibration Independent Method for Determination of the Downstream Neutral Hydrogen Density Through Ly-alpha Glow Observations

    Science.gov (United States)

    Gangopadhyay, P.; Judge, D. L.

    1996-01-01

    Our knowledge of the various heliospheric phenomena (location of the solar wind termination shock, heliopause configuration and very local interstellar medium parameters) is limited by uncertainties in the available heliospheric plasma models and by calibration uncertainties in the observing instruments. There is, thus, a strong motivation to develop model insensitive and calibration independent methods to reduce the uncertainties in the relevant heliospheric parameters. We have developed such a method to constrain the downstream neutral hydrogen density inside the heliospheric tail. In our approach we have taken advantage of the relative insensitivity of the downstream neutral hydrogen density profile to the specific plasma model adopted. We have also used the fact that the presence of an asymmetric neutral hydrogen cavity surrounding the sun, characteristic of all neutral densities models, results in a higher multiple scattering contribution to the observed glow in the downstream region than in the upstream region. This allows us to approximate the actual density profile with one which is spatially uniform for the purpose of calculating the downstream backscattered glow. Using different spatially constant density profiles, radiative transfer calculations are performed, and the radial dependence of the predicted glow is compared with the observed I/R dependence of Pioneer 10 UV data. Such a comparison bounds the large distance heliospheric neutral hydrogen density in the downstream direction to a value between 0.05 and 0.1/cc.

  13. GONAD: Genome-editing via Oviductal Nucleic Acids Delivery system: a novel microinjection independent genome engineering method in mice.

    Science.gov (United States)

    Takahashi, Gou; Gurumurthy, Channabasavaiah B; Wada, Kenta; Miura, Hiromi; Sato, Masahiro; Ohtsuka, Masato

    2015-06-22

    Microinjection is considered the gold standard technique for delivery of nucleic acids (NAs; transgenes or genome editing tools such as CRISPR/Cas9 systems) into embryos, for creating genetically modified organisms. It requires sophisticated equipment as well as well-trained and highly skilled personnel to perform the micro-injection technique. Here, we describe a novel and simple microinjection-independent technique, called Genome-editing via Oviductal Nucleic Acids Delivery (GONAD). Using GONAD, we show that NAs (e.g., eGFP mRNA or Cas9 mRNA/sgRNAs) can be effectively delivered to pre-implantation embryos within the intact mouse oviduct by a simple electroporation method, and result in the desired genetic modification in the embryos. Thus GONAD can bypass many complex steps in transgenic technology such as isolation of zygotes, microinjection of NAs into them, and their subsequent transfer to pseudo-pregnant animals. Furthermore, this method can potentially be used for genome editing in species other than mice.

  14. Dirichlet boundary conditions for arbitrary-shaped boundaries in stellarator-like magnetic fields for the Flux-Coordinate Independent method

    CERN Document Server

    Hill, Peter; Dudson, Ben

    2016-01-01

    We present a technique for handling Dirichlet boundary conditions with the Flux Coordinate Independent (FCI) parallel derivative operator with arbitrary-shaped material geometry in general 3D magnetic fields. The FCI method constructs a finite difference scheme for $\

  15. Establishment of primary bovine intestinal epithelial cell culture and clone method.

    Science.gov (United States)

    Zhan, Kang; Lin, Miao; Liu, Ming-Mei; Sui, Yang-Nan; Zhao, Guo-Qi

    2017-01-01

    The aim of this study was to establish bovine intestinal epithelial cell (BIEC) line and provide a novel clone cell method. Although various strategies of bovine cell culture and clone techniques have been reported, these methods remain not established. Here, we culture successfully primary BIECs and establish a novel clone cell method. Our result showed that BIECs could be successfully cultured and passaged about generation 5. These cellular aggregates and clusters were adherent loosely at day 2 of culture. Cell aggregates and clusters start to proliferate after approximately 4 d. The BIECs showed positive reaction against cytokeratin 18, E-cadherin, and characteristics of epithelial-like morphology. In addition, the fatty acid-binding proteins (FABPs), villin, and intestinal peptidase (IP) band were positive in BIECs. Our results suggest that the establishment of culturing and clone BIEC methods will apply to isolate and clone other primary cells. These BIECs could therefore contribute to the study of bovine intestinal nutrient absorption and regulation, immune regulation, and the pathogenesis of the bovine intestinal disease, which will provide intestinal cell model in vitro.

  16. A novel method for toxicology: in vitro culture system of a rat preantral follicle.

    Science.gov (United States)

    Wan Xuying; Zhu Jiangbo; Zhu Yuping; Xili, Ma; Liu Zhen; Wang Fei; Xu Guifeng; Zhang Tianbao

    2011-08-01

    Preantral follicle in vitro culture systems have been successfully or nearly successfully established for sheep, pig and mouse, and applied on follicle development and regulation research on reproductive biology and physiology. However, there have been few studies concerning rat preantral follicle in vitro development. The objective is to establish an in vitro culture system for rat preantral follicles which can be used for reproductive biology and toxicology research. Rat preantral follicles are mechanically separated, cultured in vitro in single follicle mode for continuous 12 days using 96-well plates, and then administrated ovulation induction. The observation on follicle development, hormone level, and ovum formation are recorded and assessed. Taking in vivo growth and in vitro maturation of oocytes group as control group, in vitro growth and maturation of oocytes group is assessed to see whether this in vitro culture method is successful. The conditions for rat follicle culture are determined based on the mouse pre-antral follicle culture. The in vitro culture system for rat preantral follicles established in this study is feasible and successful, and can serve as model for reproductive biology and toxicology research.

  17. New insights into the biogeochemistry of extremely acidic environments revealed by a combined cultivation-based and culture-independent study of two stratified pit lakes.

    Science.gov (United States)

    Falagán, Carmen; Sánchez-España, Javier; Johnson, David Barrie

    2014-01-01

    The indigenous microbial communities of two extremely acidic, metal-rich stratified pit lakes, located in the Iberian Pyrite Belt (Spain), were identified, and their roles in mediating transformations of carbon, iron, and sulfur were confirmed. A combined cultivation-based and culture-independent approach was used to elucidate microbial communities at different depths and to examine the physiologies of isolates, which included representatives of at least one novel genus and several species of acidophilic Bacteria. Phosphate availability correlated with redox transformations of iron, and this (rather than solar radiation) dictated where primary production was concentrated. Carbon fixed and released as organic compounds by acidophilic phototrophs acted as electron donors for acidophilic heterotrophic prokaryotes, many of which catalyzed the dissimilatory reduction in ferric iron; the ferrous iron generated was re-oxidized by chemolithotrophic acidophiles. Bacteria that catalyze redox transformations of sulfur were also identified, although these Bacteria appeared to be less abundant than the iron oxidizers/reducers. Primary production and microbial numbers were greatest, and biogeochemical transformation of carbon, iron, and sulfur, most intense, within a zone of c. 8-10 m depth, close to the chemocline, in both pit lakes. Archaea detected in sediments included two Thaumarchaeota clones, indicating that members of this recently described phylum can inhabit extremely acidic environments.

  18. Investigation of Legionella Contamination in Bath Water Samples by Culture, Amoebic Co-Culture, and Real-Time Quantitative PCR Methods

    Directory of Open Access Journals (Sweden)

    Akiko Edagawa

    2015-10-01

    Full Text Available We investigated Legionella contamination in bath water samples, collected from 68 bathing facilities in Japan, by culture, culture with amoebic co-culture, real-time quantitative PCR (qPCR, and real-time qPCR with amoebic co-culture. Using the conventional culture method, Legionella pneumophila was detected in 11 samples (11/68, 16.2%. Contrary to our expectation, the culture method with the amoebic co-culture technique did not increase the detection rate of Legionella (4/68, 5.9%. In contrast, a combination of the amoebic co-culture technique followed by qPCR successfully increased the detection rate (57/68, 83.8% compared with real-time qPCR alone (46/68, 67.6%. Using real-time qPCR after culture with amoebic co-culture, more than 10-fold higher bacterial numbers were observed in 30 samples (30/68, 44.1% compared with the same samples without co-culture. On the other hand, higher bacterial numbers were not observed after propagation by amoebae in 32 samples (32/68, 47.1%. Legionella was not detected in the remaining six samples (6/68, 8.8%, irrespective of the method. These results suggest that application of the amoebic co-culture technique prior to real-time qPCR may be useful for the sensitive detection of Legionella from bath water samples. Furthermore, a combination of amoebic co-culture and real-time qPCR might be useful to detect viable and virulent Legionella because their ability to invade and multiply within free-living amoebae is considered to correlate with their pathogenicity for humans. This is the first report evaluating the efficacy of the amoebic co-culture technique for detecting Legionella in bath water samples.

  19. Investigation of Legionella Contamination in Bath Water Samples by Culture, Amoebic Co-Culture, and Real-Time Quantitative PCR Methods.

    Science.gov (United States)

    Edagawa, Akiko; Kimura, Akio; Kawabuchi-Kurata, Takako; Adachi, Shinichi; Furuhata, Katsunori; Miyamoto, Hiroshi

    2015-10-19

    We investigated Legionella contamination in bath water samples, collected from 68 bathing facilities in Japan, by culture, culture with amoebic co-culture, real-time quantitative PCR (qPCR), and real-time qPCR with amoebic co-culture. Using the conventional culture method, Legionella pneumophila was detected in 11 samples (11/68, 16.2%). Contrary to our expectation, the culture method with the amoebic co-culture technique did not increase the detection rate of Legionella (4/68, 5.9%). In contrast, a combination of the amoebic co-culture technique followed by qPCR successfully increased the detection rate (57/68, 83.8%) compared with real-time qPCR alone (46/68, 67.6%). Using real-time qPCR after culture with amoebic co-culture, more than 10-fold higher bacterial numbers were observed in 30 samples (30/68, 44.1%) compared with the same samples without co-culture. On the other hand, higher bacterial numbers were not observed after propagation by amoebae in 32 samples (32/68, 47.1%). Legionella was not detected in the remaining six samples (6/68, 8.8%), irrespective of the method. These results suggest that application of the amoebic co-culture technique prior to real-time qPCR may be useful for the sensitive detection of Legionella from bath water samples. Furthermore, a combination of amoebic co-culture and real-time qPCR might be useful to detect viable and virulent Legionella because their ability to invade and multiply within free-living amoebae is considered to correlate with their pathogenicity for humans. This is the first report evaluating the efficacy of the amoebic co-culture technique for detecting Legionella in bath water samples.

  20. Comparing rapid and culture indicator bacteria methods at inland lake beaches

    Science.gov (United States)

    Francy, Donna S.; Bushon, Rebecca N.; Brady, Amie M.G.; Kephart, Christopher M.

    2013-01-01

    A rapid method, quantitative polymerase chain reaction (qPCR), for quantifying indicator bacteria in recreational waters is desirable for public health protection. We report that replacing current Escherichia coli standards with new US Environmental Protection Agency beach action values (BAVs) for enterococci by culture or qPCR may result in more advisories being posted at inland recreational lakes. In this study, concentrations of E. coli and enterococci by culture methods were compared to concentrations of Enterococcus spp. by qPCR at 3 inland lake beaches in Ohio. The E. coli and enterococci culture results were significantly related at all beaches; however, the relations between culture results and Enterococcus spp. qPCR results were not always significant and differed among beaches. All the qPCR results exceeded the new BAV for Enterococcus spp. by qPCR, whereas only 23.7% of culture results for E. coli and 79% of culture results for enterococci exceeded the current standard for E. coli or BAV for enterococci.

  1. Influence of hydroponic culture method on morphology and hydraulic conductivity of roots of honey locust.

    Science.gov (United States)

    Graves, W R

    1992-09-01

    The morphology and hydraulic conductivity of root systems of Gleditsia triacanthos L. var. inermis Willd. (honey locust) grown hydroponically in sand and solution cultures were compared. Total root system length was similar in the two cultures. However, root systems grown in solution had longer primary roots, fewer lateral roots and root hairs, and a greater distance between the tip of the primary root and the junction of the youngest secondary root and the primary root than root systems grown in sand. Hydraulic conductivities of root systems grown hydroponically for 21 or 35 days in sand or solution culture were similar. These findings show that different methods of hydroponic culture can affect root morphology without altering root resistance to water transport.

  2. Method and Apparatus for a Miniature Bioreactor System for Long-Term Cell Culture

    Science.gov (United States)

    Kleis, Stanley J. (Inventor); Geffert, Sandra K. (Inventor); Gonda, Steve R. (Inventor)

    2015-01-01

    A bioreactor and method that permits continuous and simultaneous short, moderate, or long term cell culturing of one or more cell types or tissue in a laminar flow configuration is disclosed, where the bioreactor supports at least two laminar flow zones, which are isolated by laminar flow without the need for physical barriers between the zones. The bioreactors of this invention are ideally suited for studying short, moderate and long term studies of cell cultures and the response of cell cultures to one or more stressors such as pharmaceuticals, hypoxia, pathogens, or any other stressor. The bioreactors of this invention are also ideally suited for short, moderate or long term cell culturing with periodic cell harvesting and/or medium processing for secreted cellular components.

  3. Comparison of Six Culture Methods for Salmonella Isolation from Poultry Fecal Samples

    Directory of Open Access Journals (Sweden)

    Morshed, R. (PhD

    2014-06-01

    Full Text Available Background and Objective: Salmonellosis is one of the most important food-borne bacterial zoonotic diseases worldwide, and poultry and its products are the major sources for salmonella transmission to human. Isolation of Salmonella enterica from poultry needs bacteriologic enrichment and selected cultures of fecal samples. In this study, different culture methods for the isolation of salmonella from fecal samples were compared. Material and Methods: Forty- five positive samples from infected farms and 45 negative samples from normal farms were processed using enrichment media including tetrathionate broth, selenite cistine and Rappaport-Vassiliadis. Then the samples were incubated in selective cultures, and after 24 h, their results were compared with standard method. Results: Specificity of all methods for salmonella isolation was 100%, and salmonella was not isolated from the negative samples. The highest susceptibility was related to the method in which the sample first in Selenite cistine and later in Rappaport-Vassiliadis was enriched (100%. Enrichment in Rappaport-Vassiliadis could isolate 41 salmonella from 45 positive samples (91% while the result of enrichment in tetrathionate was 6 isolates (13.3%. Conclusion: This study shows that enrichment in selenite cistine and then in Rappaport-Vassiliadis is currently the best method for isolating salmonella from fecal samples of poultry. Key words: Salmonella; Bacteriologic Culture; Diagnosis; Isolation; Enrichment; Poultry

  4. The Efficient Method for Simultaneous Monitoring of the Culturable as Well as Nonculturable Airborne Microorganisms

    OpenAIRE

    2013-01-01

    Cultivation-based microbiological methods are a gold standard for monitoring of airborne micro-organisms to determine the occupational exposure levels or transmission paths of a particular infectious agent. Some highly contagious microorganisms are not easily culturable but it is becoming evident that cultivation and molecular methods are complementary and in these cases highly relevant. We report a simple and efficient method for sampling and analyzing airborne bacteria with an impactor-type...

  5. Detection of Food Hazards in Foods: Comparison of Real Time Polymerase Chain Reaction and Cultural Methods.

    Science.gov (United States)

    Bonilauri, Paolo; Bardasi, Lia; Leonelli, Roberto; Ramini, Mattia; Luppi, Andrea; Giacometti, Federica; Merialdi, Giuseppe

    2016-01-18

    Foodstuffs should not contain microorganisms or their toxins or metabolites in quantities suggesting an unacceptable risk for human health. The detection of food hazards in foods is performed by several tests that produce results dependent on the analytical method used: an analytical reference method, defined as standard, is associated with each microbiological criterion laid down in Regulation 2073/2005/EC, but, analytical methods other than the reference ones, in particular more rapid methods, could be used. Combined screening methods performed by real time-polymerase chain reaction (RT-PCR) are currently validated as alternative methods according to the ISO 16140:2003 and certified by the Association Française de Normalisation. However, the positive results obtained with these alternative methods, the investigated molecular relations that resulted positive have to be confirmed with cultural methods using the same enrichment media in which the molecular screening was performed. Since it is necessary to assess if these testing schemes provide equivalent guarantees of food safety, the aim of this retrospective study is to analyse the data collected, from 2012 to 2014 by Emilia Romagna Region in the field of Piano Regionale Alimenti (Food Regional Plan) during official controls monitoring food samples of animal and other than animal origin. Records performed by combined methods of molecular screening of Salmonella spp., Listeria monocytogenes and thermophilic Campylobacter and cultural confirmation results were gathered together and the results were compared in order to assess the sensitivity of the methods. A total of 10,604 food samples were considered in this study: the comparison of the data revealed that the RT-PCR method detected Salmonella, L. monocytogenes, and thermophilic Campylobacter in 2.18, 3.85 and 3.73% of the samples, respectively, whereas by using cultural method these pathogens were isolated in 0.43, 1.57 and 1.57% of samples, respectively. In

  6. Detection of food hazards in foods: comparison of real time polymerase chain reaction and cultural methods

    Directory of Open Access Journals (Sweden)

    Paolo Bonilauri

    2016-01-01

    Full Text Available Foodstuffs should not contain microorganisms or their toxins or metabolites in quantities suggesting an unacceptable risk for human health. The detection of food hazards in foods is performed by several tests that produce results dependent on the analytical method used: an analytical reference method, defined as standard, is associated with each microbiological criterion laid down in Regulation 2073/2005/EC, but, analytical methods other than the reference ones, in particular more rapid methods, could be used. Combined screening methods performed by real time-polymerase chain reaction (RT-PCR are currently validated as alternative methods according to the ISO 16140:2003 and certified by the Association Française de Normalisation. However, the positive results obtained with these alternative methods, the investigated molecular relations that resulted positive have to be confirmed with cultural methods using the same enrichment media in which the molecular screening was performed. Since it is necessary to assess if these testing schemes provide equivalent guarantees of food safety, the aim of this retrospective study is to analyse the data collected, from 2012 to 2014 by Emilia Romagna Region in the field of Piano Regionale Alimenti (Food Regional Plan during official controls monitoring food samples of animal and other than animal origin. Records performed by combined methods of molecular screening of Salmonella spp., Listeria monocytogenes and thermophilic Campylobacter and cultural confirmation results were gathered together and the results were compared in order to assess the sensitivity of the methods. A total of 10,604 food samples were considered in this study: the comparison of the data revealed that the RT-PCR method detected Salmonella, L. monocytogenes, and thermophilic Campylobacter in 2.18, 3.85 and 3.73% of the samples, respectively, whereas by using cultural method these pathogens were isolated in 0.43, 1.57 and 1.57% of samples

  7. A hybrid microfluidic-vacuum device for direct interfacing with conventional cell culture methods.

    Science.gov (United States)

    Chung, Bong Geun; Park, Jeong Won; Hu, Jia Sheng; Huang, Carlos; Monuki, Edwin S; Jeon, Noo Li

    2007-09-20

    Microfluidics is an enabling technology with a number of advantages over traditional tissue culture methods when precise control of cellular microenvironment is required. However, there are a number of practical and technical limitations that impede wider implementation in routine biomedical research. Specialized equipment and protocols required for fabrication and setting up microfluidic experiments present hurdles for routine use by most biology laboratories. We have developed and validated a novel microfluidic device that can directly interface with conventional tissue culture methods to generate and maintain controlled soluble environments in a Petri dish. It incorporates separate sets of fluidic channels and vacuum networks on a single device that allows reversible application of microfluidic gradients onto wet cell culture surfaces. Stable, precise concentration gradients of soluble factors were generated using simple microfluidic channels that were attached to a perfusion system. We successfully demonstrated real-time optical live/dead cell imaging of neural stem cells exposed to a hydrogen peroxide gradient and chemotaxis of metastatic breast cancer cells in a growth factor gradient. This paper describes the design and application of a versatile microfluidic device that can directly interface with conventional cell culture methods. This platform provides a simple yet versatile tool for incorporating the advantages of a microfluidic approach to biological assays without changing established tissue culture protocols.

  8. A hybrid microfluidic-vacuum device for direct interfacing with conventional cell culture methods

    Directory of Open Access Journals (Sweden)

    Monuki Edwin S

    2007-09-01

    Full Text Available Abstract Background Microfluidics is an enabling technology with a number of advantages over traditional tissue culture methods when precise control of cellular microenvironment is required. However, there are a number of practical and technical limitations that impede wider implementation in routine biomedical research. Specialized equipment and protocols required for fabrication and setting up microfluidic experiments present hurdles for routine use by most biology laboratories. Results We have developed and validated a novel microfluidic device that can directly interface with conventional tissue culture methods to generate and maintain controlled soluble environments in a Petri dish. It incorporates separate sets of fluidic channels and vacuum networks on a single device that allows reversible application of microfluidic gradients onto wet cell culture surfaces. Stable, precise concentration gradients of soluble factors were generated using simple microfluidic channels that were attached to a perfusion system. We successfully demonstrated real-time optical live/dead cell imaging of neural stem cells exposed to a hydrogen peroxide gradient and chemotaxis of metastatic breast cancer cells in a growth factor gradient. Conclusion This paper describes the design and application of a versatile microfluidic device that can directly interface with conventional cell culture methods. This platform provides a simple yet versatile tool for incorporating the advantages of a microfluidic approach to biological assays without changing established tissue culture protocols.

  9. Assessment of central venous catheter-associated infections using semi-quantitative or quantitative culture methods

    Directory of Open Access Journals (Sweden)

    E. L. Pizzolitto

    2009-01-01

    Full Text Available

    Semiquantitative (Maki and quantitative (Brun- Buisson culture techniques were employed in the diagnosis of catheter-related bloodstream infections (CRBSI in patients who have a short-term central venous catheter (inserted for 30 days. The diagnosis of CRBSI was based on the results of semiquantitative and quantitative culture of material from the removed catheters. Catheter tips (118 from 100 patients were evaluated by both methods. Semiquantitative analysis revealed 34 catheters (28.8% colonized by ≥15 colonyforming units (cfu, while quantitative cultures (34 catheters, 28.8% showed the growth of ≥103 cfu/mL. Bacteremia was confirmed in four patients by isolating microorganisms of identical species from both catheters and blood samples. Using the semiquantitative culture technique on short-term central venous catheter tips, we have shown that with a cut-off level of ≥15 cfu, the technique had 100.0% sensitivity, specificity of 68.4%, 25.0% positive predictive value (PPV and 100.0% negative predictive value (NPV, efficiency of 71.4% and a prevalence of 9.5%. The quantitative method, with a cut-off limit of ≥103 cfu/mL, gave identical values: the sensitivity was 100.0%, specificity 68.4%, positive predictive value (PPV 25.0%, negative predictive value (NPV 100.0%, efficiency 71.4% and prevalence 9.5%. We concluded that the semiquantitative and quantitative culture methods, evaluated in parallel, for the first time in Brazil, have similar sensitivity and specificity. Keywords: central venous catheter; semi-quantitative culture; quantitative culture; catheter-related bacteremia.

  10. Comparison of Uriswab to alternative methods for urine culture collection and transport: confirmation of standard culture methodology for investigation of urinary tract infections.

    Science.gov (United States)

    Rennie, Robert P; Turnbull, Lee-Ann; Gauchier-Pitts, Kaylee; Bennett, Tracy; Dyrland, Debbie; Blonski, Susan

    2016-08-01

    The ability to isolate and identify causative agents of urinary tract infections relies primarily on the quality of the urine sample that is submitted to the microbiology. The most important factors are the method of collection, the maintenance of viability of the potential pathogens during transport, and standardization of the culturing of the urine sample. This report is a composite of several investigations comparing collection and transport on urine culture paddles, with a preservative urine sponge (Uriswab), and a comparison of Uriswab with the BD preservative transport tube as methods of preservation of urinary pathogens. Primary studies showed that Uriswab maintained significantly more urinary pathogens than the urine culture paddle with fewer mixed or contaminated cultures. The two preservative transport systems were comparable for maintenance of viability of the pathogens, but there were fewer mixed cultures when samples were collected with Uriswab. This study confirms the importance of a standard volume of 1 μL of urine for culture.

  11. cultural

    Directory of Open Access Journals (Sweden)

    Irene Kreutz

    2006-01-01

    Full Text Available Es un estudio cualitativo que adoptó como referencial teorico-motodológico la antropología y la etnografía. Presenta las experiencias vivenciadas por mujeres de una comunidad en el proceso salud-enfermedad, con el objetivo de comprender los determinantes sócio-culturales e históricos de las prácticas de prevención y tratamiento adoptados por el grupo cultural por medio de la entrevista semi-estructurada. Los temas que emergieron fueron: la relación entre la alimentación y lo proceso salud-enfermedad, las relaciones con el sistema de salud oficial y el proceso salud-enfermedad y lo sobrenatural. Los dados revelaron que los moradores de la comunidad investigada tienen un modo particular de explicar sus procedimientos terapéuticos. Consideramos que es papel de los profesionales de la salud en sus prácticas, la adopción de abordajes o enfoques que consideren al individuo en su dimensión sócio-cultural e histórica, considerando la enorme diversidad cultural en nuestro país.

  12. Transferring methods to teach business administration from one cultural context to another

    Directory of Open Access Journals (Sweden)

    Marie Catalo

    2015-12-01

    Full Text Available What happens when a teaching method is transferred from one cultural context to another? In this article we investigate this question by looking at how Computer Based Simulations (CBS were transposed from a French context to an Egyptian one. In this article we demonstrate, through the case of Egypt, how culture and the characteristics of the school system impact learning abilities. We describe what happens when Egyptian students are confronted with learning modes they have not encountered prior to University, in the context of an Egyptian-French dual-degree programme in business administration and business informatics. We show that the transfer of CBS as a teaching method revealed cultural differences between French and Egyptian students. As a consequence the teaching objectives of CBS were redefined in order to take the Egyptian context into account.

  13. [Investigation of the presence of Blastocystis spp. in stool samples with microscopic, culture and molecular methods].

    Science.gov (United States)

    Adıyaman Korkmaz, Gülcan; Doğruman Al, Funda; Mumcuoğlu, İpek

    2015-01-01

    Blastocystis species are enteric protozoa frequently detected in human and animals. Seventeen subtypes (STs) have now been identified, nine of them isolating from humans. The pleomorphic structure and genetic diversity of Blastocystis spp. and the absence of standardized diagnostic methods complicate the evaluation of current data. Microscopic methods such as native-lugol and trichrome staining are most frequently used methods in routine diagnosis, while culture and molecular methods are preferred for research purposes. The aims of this study were to investigate the presence of Blastocystis spp. in the stool samples of patients with gastrointestinal complaints by microscopic and culture methods, and to detect the subtypes of isolates by polymerase chain reaction (PCR). A total of 350 stool samples collected from patients with diarrhea (n= 157) and without diarrhea (n= 193) were included in the study. Presence of Blastocystis spp. in the samples were investigated by native-lugol examination, trichrome staining and direct fluorescent antibody (DFA) methods. Ringer's solution containing 10% horse serum and 0.05% asparagine was used for cultivation. The cultures were evaluated after 3-4 days of incubation at 37°C by microscopic examination. The subtypes of Blastocystis spp. strains isolated from the cultures have been identified by PCR using sequence-tagged site primers. A total of 66 (19%) stool samples, of them 26 (16.6%) were from diarrheal and 40 (21%) from non-diarrheal cases, yielded Blastocystis sp. growth in culture. Among the evaluated samples, 12% (42/350) were found positive with native-lugol examination, 17% (58/350) with trichrome staining, and 19% (66/350) with DFA method. The agreement of culture and native-lugol method was estimated as strong (κ= 0.752), while it was very strong between culture with trichrome staining and DFA methods (κ= 0.922 and κ= 1.00, respectively). When the culture was accepted as reference method, the sensitivity and

  14. Room for improvement? Leadership, innovation culture and uptake of quality improvement methods in general practice.

    Science.gov (United States)

    Apekey, Tanefa A; McSorley, Gerry; Tilling, Michelle; Siriwardena, A Niroshan

    2011-04-01

    Leadership and innovation are currently seen as essential elements for the development and maintenance of high-quality care. Little is known about the relationship between leadership and culture of innovation and the extent to which quality improvement methods are used in general practice. This study aimed to assess the relationship between leadership behaviour, culture of innovation and adoption of quality improvement methods in general practice. Self-administered postal questionnaires were sent to general practitioner quality improvement leads in one county in the UK between June and December 2007. The questionnaire consisted of background information, a 12-item scale to assess leadership behaviour, a seven-dimension self-rating scale for culture of innovation and questions on current use of quality improvement tools and techniques. Sixty-three completed questionnaires (62%) were returned. Leadership behaviours were not commonly reported. Most practices reported a positive culture of innovation, featuring relationship most strongly, followed by targets and information but rated lower on other dimensions of rewards, risk and resources. There was a significant positive correlation between leadership behaviour and the culture of innovation (r = 0.57; P quality improvement methods were not adopted by most participating practices. Leadership behaviours were infrequently reported and this was associated with a limited culture of innovation in participating general practices. There was little use of quality improvement methods beyond clinical and significant event audit. Practices need support to enhance leadership skills, encourage innovation and develop quality improvement skills if improvements in health care are to accelerate. © 2010 Blackwell Publishing Ltd.

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

    Science.gov (United States)

    Li, G; Li, M; Ma, J

    1992-02-01

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

  16. X-ray and synchrotron methods in studies of cultural heritage sites

    Science.gov (United States)

    Koval'chuk, M. V.; Yatsishina, E. B.; Blagov, A. E.; Tereshchenko, E. Yu.; Prosekov, P. A.; Dyakova, Yu. A.

    2016-09-01

    X-ray and synchrotron methods that are most widely used in studies of cultural heritage objects (including archaeological sites)—X-ray diffraction analysis, X-ray spectroscopy, and visualization techniques— have been considered. The reported examples show high efficiency and informativeness of natural science studies when solving most diverse problems of archaeology, history, the study of art, museology, etc.

  17. The Effects of Culture and Social Class on Client Preference for Counseling Methods.

    Science.gov (United States)

    Anderson, John W.

    1983-01-01

    Surveyed the preferences of 66 Black, White, and Mexican-American college students for directive and nondirective counseling analogs to determine the impact of culture and social class in counseling. Results showed Blacks generally scored the two methods exactly opposite their White and Mexican-American counterparts. (JAC)

  18. Learning as Researchers and Teachers: The Development of a Pedagogical Culture for Social Science Research Methods?

    Science.gov (United States)

    Kilburn, Daniel; Nind, Melanie; Wiles, Rose

    2014-01-01

    In light of calls to improve the capacity for social science research within UK higher education, this article explores the possibilities for an emerging pedagogy for research methods. A lack of pedagogical culture in this field has been identified by previous studies. In response, we examine pedagogical literature surrounding approaches for…

  19. Anthropological Methods of Formation of University Students' Spiritual and Moral Culture

    Science.gov (United States)

    Kurbanov, Rashad A.; Nikonova, Elina I.; Gurbanov, Ramin A.; Svechnikova, Natalia V.; Tumarov, Konstantin B.; Marin, Evgeniy M.

    2016-01-01

    The relevance of the study is reasoned by the increasing complexity of life activity in modern society, which results in distortion of the moral and value criteria and norms. The purpose of the article is to reveal anthropological methods of formation of university students' spiritual and moral culture. The leading approach to the study is the…

  20. Anthropology and International Business Research Methods in DBA Teaching: Frameworks for Cultural Awareness.

    Science.gov (United States)

    Whiteley, Alma

    2001-01-01

    Discusses the rationale for introducing anthropology into a doctoral-level international business research methods course. Describes three anthropological frameworks designed for the course: a cultural awareness model adapted from G. Morgan's (1980) idea of paradigmatic orthodoxy; key organizing principles; and a mapping model allowing researchers…

  1. Learning as Researchers and Teachers: The Development of a Pedagogical Culture for Social Science Research Methods?

    Science.gov (United States)

    Kilburn, Daniel; Nind, Melanie; Wiles, Rose

    2014-01-01

    In light of calls to improve the capacity for social science research within UK higher education, this article explores the possibilities for an emerging pedagogy for research methods. A lack of pedagogical culture in this field has been identified by previous studies. In response, we examine pedagogical literature surrounding approaches for…

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

    Science.gov (United States)

    Grant, Natalie S.; Bolin, Brien L.

    2016-01-01

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

  3. Induced Pluripotent Stem (iPS) Cell Culture Methods and Induction of Differentiation into Endothelial Cells

    Science.gov (United States)

    Chatterjee, Ishita; Li, Fei; Kohler, Erin E.; Rehman, Jalees; Malik, Asrar B.; Wary, Kishore K.

    2015-01-01

    Summary The studies of stem cell behavior and differentiation in a developmental context is complex, time-consuming and expensive, and for this reason, cell culture remains a method of choice for developmental and regenerative biology and mechanistic studies. Similar to ES cells, iPS cells have the ability to differentiate into endothelial cells (ECs), and the route for differentiation appears to mimic the developmental process that occurs during the formation of an embryo. Traditional EC induction methods from embryonic stem (ES) cells rely mostly on the formation the embryoid body (EB), which employs feeder or feeder-free conditions in the presence or absence of supporting cells. Similar to ES cells, iPS cells can be cultured in feeder-layer or feeder-free conditions. Here, we describe the iPS cell culture methods and induction differentiation of these cells into ECs. We use anti-mouse Flk1 and anti-mouse VE-cadherin to isolate and characterize mouse ECs, because these antibodies are commercially available and their use has been described in the literature, including by our group. The ECs produced by this method have been used by our laboratory, and we have demonstrated their in vivo potential. We also discuss how iPS cells differ in their ability to differentiate into endothelial cells in culture. PMID:25687301

  4. Induced Pluripotent Stem (iPS) Cell Culture Methods and Induction of Differentiation into Endothelial Cells.

    Science.gov (United States)

    Chatterjee, Ishita; Li, Fei; Kohler, Erin E; Rehman, Jalees; Malik, Asrar B; Wary, Kishore K

    2016-01-01

    The study of stem cell behavior and differentiation in a developmental context is complex, time-consuming, and expensive, and for this reason, cell culture remains a method of choice for developmental and regenerative biology and mechanistic studies. Similar to ES cells, iPS cells have the ability to differentiate into endothelial cells (ECs), and the route for differentiation appears to mimic the developmental process that occurs during the formation of an embryo. Traditional EC induction methods from embryonic stem (ES) cells rely mostly on the formation of embryoid body (EB), which employs feeder or feeder-free conditions in the presence or absence of supporting cells. Similar to ES cells, iPS cells can be cultured in feeder layer or feeder-free conditions. Here, we describe the iPS cell culture methods and induction differentiation of these cells into ECs. We use anti-mouse Flk1 and anti-mouse VE-cadherin to isolate and characterize mouse ECs, because these antibodies are commercially available and their use has been described in the literature, including by our group. The ECs produced by this method have been used by our laboratory, and we have demonstrated their in vivo potential. We also discuss how iPS cells differ in their ability to differentiate into endothelial cells in culture.

  5. Children's activities and their meanings for parents: a mixed-methods study in six Western cultures.

    Science.gov (United States)

    Harkness, Sara; Zylicz, Piotr Olaf; Super, Charles M; Welles-Nyström, Barbara; Bermúdez, Moisés Ríos; Bonichini, Sabrina; Moscardino, Ughetta; Mavridis, Caroline Johnston

    2011-12-01

    Theoretical perspectives and research in sociology, anthropology, sociolinguistics, and cultural psychology converge in recognizing the significance of children's time spent in various activities, especially in the family context. Knowing how children's time is deployed, however, only gives us a partial answer to how children acquire competence; the other part must take into account the culturally constructed meanings of activities, from the perspective of those who organize and direct children's daily lives. In this article, we report on a study of children's routine daily activities and on the meanings that parents attribute to them in six Western middle-class cultural communities located in Italy, The Netherlands, Poland, Spain, Sweden, and the United States (N = 183). Using week-long time diaries kept by parents, we first demonstrate similarities as well as significant differences in children's daily routines across the cultural samples. We then present brief vignettes--"a day in the life" --of children from each sample. Parent interviews were coded for themes in the meanings attributed to various activities. Excerpts from parent interviews, focusing on four major activities (meals, family time, play, school- or developmentally related activities), are presented to illustrate how cultural meanings and themes are woven into parents' organization and understanding of their children's daily lives. The results of this mixed-method approach provide a more reliable and nuanced picture of children's and families' daily lives than could be derived from either method alone.

  6. An investigation of Helicobacter pylori using culture, histopathological and serological examination methods and its antimicrobial sensitivities.

    Science.gov (United States)

    Inan, Asuman; Gulsun, Serda; Guveli, Hakan; Tascioglu, Jale; Goktas, Pasa

    2005-04-01

    In this study, the determination of Helicobacter pylori (H. pylori) by culture, histopathological and serological methods in cases of endoscopically diagnosed as duodenitis and duodenal ulcer (DU), a comparison of their relative advantages, and its antibiotic sensitivities were investigated. Helicobacter pylori was investigated using 3 methods (culture, histopathological and serological examination) in 50 patients (25 diagnosed with duodenitis and 25 with DU) at the Department of Gastroenterohepatology, Istanbul Haydarpasa Numune Hospital, Turkey between December 2000 and February 2001. An investigation into its antibiotic sensitivities to amoxicillin, clarithromycin, metronidazole and azithromycin by disc diffusion methods and to amoxicillin and clarithromycin by E-test were investigated. Helicobacter pylori bacteria were observed in Gram stained preparates prepared from biopsy material in 34 out of 50 patients (68%), and were able to be produced in active culture in all these cases. Histopathological examination revealed the presence of H. pylori in 80% cases of DU and 60% cases of duodenitis; anti-CagA(IgG) was positively determined in 88% DU cases and in 60% duodenitis cases. There was a significant difference between the 2 groups in terms of diagnosis by histopathological and serological methods. The difference between the 2 groups produced in active culture in 84% cases of DU cases and 52% of duodenitis was statistically significant (p=0.0322). Using the E-test and disc diffusion methods, 8.8% of the strains that reproduced in culture were resistant to and 91.2% were sensitive to clarithromycin. All strains were determined to be sensitive to amoxicillin: 17.6% of the strains were determined to be resistant to metronidazole, 11.7% to azithromycin. It was observed that Gram staining is a rapid and reliable method of pre-diagnosis for H. pylori; that histopathological examination methods are of considerable importance in diagnosis; and that the investigation of

  7. Statins reduce amyloid β-peptide production by modulating amyloid precursor protein maturation and phosphorylation through a cholesterol-independent mechanism in cultured neurons.

    Science.gov (United States)

    Hosaka, Ai; Araki, Wataru; Oda, Akiko; Tomidokoro, Yasushi; Tamaoka, Akira

    2013-03-01

    Statins, 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors, have been reported to attenuate amyloid-β peptide (Aβ) production in various cellular models. However, the mechanisms by which statins affect neuronal Aβ production have not yet been clarified. Here, we investigated this issue in rat primary cortical neurons using two statins, pitavastatin (PV) and atorvastatin (AV). Treatment of neurons with 0.2-2.5 μM PV or AV for 4 days induced a concentration- and time-dependent reduction in the secretion of both Aβ40 and Aβ42. Moreover, Western blot analyses of cell lysates showed that treatment with PV or AV significantly reduced expression levels of the mature form of amyloid precursor protein (APP) and Thr668-phosphorylated APP (P-APP), but not immature form of APP; the decreases in P-APP levels were more notable than those of mature APP levels. The statin treatment did not alter expression of BACE1 (β-site APP-cleaving enzyme 1) or γ-secretase complex proteins (presenilin 1, nicastrin, APH-1, and PEN-2). In neurons overexpressing APP via recombinant adenoviruses, PV or AV similarly reduced Aβ secretion and the levels of mature APP and P-APP. Statins also markedly reduced cellular cholesterol content in neurons in a concentration-dependent manner. Co-treatment with mevalonate reversed the statin-induced decreases in Aβ secretion and mature APP and P-APP levels, whereas co-treatment with cholesterol did not, despite recovery of cellular cholesterol levels. Finally, cell-surface biotinylation experiments revealed that both statins significantly reduced the levels of cell-surface P-APP without changing those of cell surface mature APP. These results suggest that statins reduce Aβ production by selectively modulating APP maturation and phosphorylation through a mechanism independent of cholesterol reduction in cultured neurons.

  8. Optimizing seeding and culture methods to engineer smooth muscle tissue on biodegradable polymer matrices.

    Science.gov (United States)

    Kim, B S; Putnam, A J; Kulik, T J; Mooney, D J

    1998-01-05

    The engineering of functional smooth muscle (SM) tissue is critical if one hopes to successfully replace the large number of tissues containing an SM component with engineered equivalents. This study reports on the effects of SM cell (SMC) seeding and culture conditions on the cellularity and composition of SM tissues engineered using biodegradable matrices (5 x 5 mm, 2-mm thick) of polyglycolic acid (PGA) fibers. Cells were seeded by injecting a cell suspension into polymer matrices in tissue culture dishes (static seeding), by stirring polymer matrices and a cell suspension in spinner flasks (stirred seeding), or by agitating polymer matrices and a cell suspension in tubes with an orbital shaker (agitated seeding). The density of SMCs adherent to these matrices was a function of cell concentration in the seeding solution, but under all conditions a larger number (approximately 1 order of magnitude) and more uniform distribution of SMCs adherent to the matrices were obtained with dynamic versus static seeding methods. The dynamic seeding methods, as compared to the static method, also ultimately resulted in new tissues that had a higher cellularity, more uniform cell distribution, and greater elastin deposition. The effects of culture conditions were next studied by culturing cell-polymer constructs in a stirred bioreactor versus static culture conditions. The stirred culture of SMC-seeded polymer matrices resulted in tissues with a cell density of 6.4 +/- 0.8 x 10(8) cells/cm3 after 5 weeks, compared to 2.0 +/- 1.1 x 10(8) cells/cm3 with static culture. The elastin and collagen synthesis rates and deposition within the engineered tissues were also increased by culture in the bioreactors. The elastin content after 5-week culture in the stirred bioreactor was 24 +/- 3%, and both the elastin content and the cellularity of these tissues are comparable to those of native SM tissue. New tissues were also created in vivo when dynamically seeded polymer matrices were

  9. The truncated polynomial expansion Monte Carlo method for fermion systems coupled to classical fields: a model independent implementation

    Science.gov (United States)

    Alvarez, G.; Şen, C.; Furukawa, N.; Motome, Y.; Dagotto, E.

    2005-05-01

    A software library is presented for the polynomial expansion method (PEM) of the density of states (DOS) introduced in [Y. Motome, N. Furukawa, J. Phys. Soc. Japan 68 (1999) 3853; N. Furukawa, Y. Motome, H. Nakata, Comput. Phys. Comm. 142 (2001) 410]. The library provides all necessary functions for the use of the PEM and its truncated version (TPEM) in a model independent way. The PEM/TPEM replaces the exact diagonalization of the one electron sector in models for fermions coupled to classical fields. The computational cost of the algorithm is O(N)—with N the number of lattice sites—for the TPEM [N. Furukawa, Y. Motome, J. Phys. Soc. Japan 73 (2004) 1482] which should be contrasted with the computational cost of the diagonalization technique that scales as O(N). The method is applied for the first time to a double exchange model with finite Hund coupling and also to diluted spin-fermion models. Program summaryTitle of library:TPEM Catalogue identifier: ADVK Program summary URL:http://cpc.cs.qub.ac.uk/summaries/ADVK Program obtainable from: CPC Program Library, Queen's University of Belfast, N. Ireland No. of lines in distributed program, including test data, etc.: 1707 No. of bytes in distributed program, including test data, etc.: 13 644 Distribution format:tar.gz Operating system:Linux, UNIX Number of files:4 plus 1 test program Programming language used:C Computer:PC Nature of the physical problem:The study of correlated electrons coupled to classical fields appears in the treatment of many materials of much current interest in condensed matter theory, e.g., manganites, diluted magnetic semiconductors and high temperature superconductors among others. Method of solution: Typically an exact diagonalization of the electronic sector is performed in this type of models for each configuration of classical fields, which are integrated using a classical Monte Carlo algorithm. A polynomial expansion of the density of states is able to replace the exact

  10. Radiation-Independent Distal Locking Screw Insertion Using Dual Nail Insertion Handle: Is it a Reliable Method?

    Directory of Open Access Journals (Sweden)

    Adel Abdel Azim Foda

    2017-06-01

    Conclusion: This technique is not reliable enough to replace the classic radiation-dependent free-hand technique. Further development of this idea is needed to get a perfect radiation-independent distal locking technique.

  11. A procedure to evaluate the efficiency of surface sterilization methods in culture-independent fungal endophyte studies.

    Science.gov (United States)

    Burgdorf, R J; Laing, M D; Morris, C D; Jamal-Ally, S F

    2014-01-01

    Extraneous DNA interferes with PCR studies of endophytic fungi. A procedure was developed with which to evaluate the removal of extraneous DNA. Wheat (Triticum aestivum) leaves were sprayed with Saccharomyces cerevisiae and then subjected to physical and chemical surface treatments. The fungal ITS1 products were amplified from whole tissue DNA extractions. ANOVA was performed on the DNA bands representing S. cerevisiae on the agarose gel. Band profile comparisons using permutational multivariate ANOVA (PERMANOVA) and non-metric multidimensional scaling (NMDS) were performed on DGGE gel data, and band numbers were compared between treatments. Leaf surfaces were viewed under variable pressure scanning electron microscopy (VPSEM). Yeast band analysis of the agarose gel showed that there was no significant difference in the mean band DNA quantity after physical and chemical treatments, but they both differed significantly (p sterilization techniques in DNA-based fungal endophyte studies.

  12. Validation of the H2S method to detect bacteria of fecal origin by cultured and molecular methods.

    Science.gov (United States)

    McMahan, Lanakila; Devine, Anthony A; Grunden, Amy M; Sobsey, Mark D

    2011-12-01

    Using biochemical and molecular methods, this research determined whether or not the H(2)S test did correctly identify sewage-contaminated waters by being the first to use culturing and molecular methods to identify the types and numbers of fecal indicator organisms, pathogens, and other microbes present in sewage samples with positive H(2)S test results. For the culture-based method, samples were analyzed for the presence of fecal bacteria by spread plating the sewage sample onto differential and selective media for Aeromonas spp., Escherichia coli, sulfite-reducing clostridia, H(2)S-producing bacteria, and Salmonella/Shigella spp. The isolates were then: (1) tested to determine whether they were H(2)S-producing organisms and (2) identified to the genus and species level using biochemical methods. The molecular method used to characterize the microbial populations of select samples was terminal restriction fragment length polymorphisms. These experiments on sewage provided evidence that positive H(2)S tests consistently contained fecal bacteria and pathogens. There were strong relationships of agreement between the organisms identified by both methods tested. This study is an important advance in microbial water quality detection since it is focused on the evaluation of a novel, low-cost, water microbiology test that has the potential to provide millions of people worldwide access to water quality detection technology. Of prime consideration in evaluating water quality tests is the determination of the test's accuracy and specificity, and this article is a fundamental step in providing that information.

  13. Assessment of microbial diversity under arid plants by culture ...

    African Journals Online (AJOL)

    Dr. R. K. Jain

    2013-10-02

    Oct 2, 2013 ... Both culture- dependent and culture-independent methods indicated that in arid crops, ... on analysis of DNA allow investigation of this potential. .... Addition of anionic detergent, SDS along with CTAB yielded maximum DNA.

  14. Comparison of Real-time PCR method and blood culture in diagnosis of septicemia

    Directory of Open Access Journals (Sweden)

    Ali Gholami

    2016-02-01

    Full Text Available Background: Bloodstream infections (BSI have a high incidence and high mortality in the worldwide. The mortality rate is variable between 20-70%. Therefore, early and timely detection of BSI agent in clinical laboratories is necessary. The aim of this study was to determine an efficient diagnostic tool to septicemia in accompany of blood culture method by Real-time PCR (using panbacterial 23S rRNA gene. Methods: This cross-sectional study was conducted in two analytical and clinical stages in Hamadan University of Medical Sciences, Iran, from October 2014 to June 2015. In analytical stage, sensitivity (by serial dilution from 104 to 1 CFU/ml and specificity of the primer were evaluated with the Staphylococcus aureus (as Gram positive indicator bacteria and Escherichia coli (as Gram-negative indicator bacteria, human genome (from Hella cell culture, Candida albicans yeast and Aspergillus fumigatus fungus. In clinical stage, 121 blood samples were collected from patients suspected to sepsis in intensive care unit (ICU from Hamadan University Hospitals. Finally, the results of Real-time PCR and blood culture methods were compared. Results: The Real-time PCR showed a sensitivity ranging from 2 to 10 target copies per reaction to the whole blood for Escherichia coli and Staphylococcus aureus respectively. The specificity of this method was evaluated and no false positive amplification was identified. 57.85% (70 cases of the samples were positive by Real-time PCR and 13.22% (16 cases of the samples were positive by blood culture. However, none of the cases that were positive by blood culture were negative in Real-time PCR. As well as, 44.62% (54 cases of cases were positive by Real-time PCR but blood culture showed no bacteria in the samples, and 42.15% (51 cases were negative by both methods. Correlation or agreement of Kappa was 0.20, that indicating poor agreement between the two methods. Conclusion: Real-time PCR is more sensitive than blood

  15. [Comparison of direct microscopy, culture and polymerase chain reaction methods for the diagnosis of cutaneous leishmaniasis].

    Science.gov (United States)

    Ertabaklar, Hatice; Özlem Çalışkan, Serçin; Boduç, Erengül; Ertuğ, Sema

    2015-01-01

    Cutaneous leishmaniasis (CL) is an endemic disease especially in Southeastern Anatolia of Turkey and recently shows a trend for spread to other regions of the country including the Aegean region. The diagnosis of CL is based on combined evaluation of epidemiological data with the clinical symptoms and laboratory findings. Direct microscopic examination and culture methods are mainly used in the routine diagnosis of CL, while molecular methods are mainly used for research. The aim of this study was to detect the presence of Leishmania spp. in samples obtained from CL-suspected patients by using direct microscopy, culture and polymerase chain reaction (PCR) methods and to compare the results. A total of 55 patients who were admitted to Parasitology Laboratory of Adnan Menderes University Hospital, Aydin (located at Aegean region in Turkey), between 2012-2014 were included in the study. Smear preparations from the skin lesions of cases were fixed and stained with Giemsa, and the presence of amastigote forms were evaluated by direct microscopy. NNN medium was used for the cultivation of samples. Total genomic DNA of Leishmania from the samples were extracted with a commercial kit (NucleoSpin Tissue(®) Kit, Macherey-Nagel, Germany) and PCR was performed by using 13A and 13B primers to amplify the 116 base pair fragment of Leishmania spp. specific kinetoplast DNA. Amastigotes were observed in 29 (53%) of the 55 samples by direct microscopy, promastigotes were detected among 34 (62%) samples in culture, and parasite-specific amplicons were revealed in 30 (55%) samples by PCR. All assays were positive in 24 patients while in 18 patients all of the tests yielded negative results. Thirty-seven (67%) out of 55 cases were diagnosed as CL when reactivity in at least one of these three methods were considered as positive. Accordingly the positivity rates of the methods were 78.4% (29/37) for direct microscopy, 92% (34/37) for culture and 81.1% (30/37) for PCR in CL

  16. METHOD FOR INDUCING SPORULATION OF PURE CULTURES OF THE MYXOMYCETE PHYSARUM POLYCEPHALUM

    Science.gov (United States)

    Daniel, John W.; Rusch, Harold P.

    1962-01-01

    Daniel, John W. (University of Wisconsin, Madison) and Harold P. Rusch. Method for inducing sporulation of pure cultures of the myxomycete Physarum polycephalum. J. Bacteriol. 83:234–240. 1962.—Techniques for inducing sporulation of pure cultures of Physarum polycephalum are described. Small plasmodia grown in agitated culture were harvested, allowed to fuse into large plasmodia (1.3 g wet weight each), and, after incubation in the dark on a salts medium, exposed to light for 2 hr. The ensuing formation of sporangia containing spores was complete after 12 to 16 hr. The obligatory conditions for sporulation are: (i) an optimal growth age occurring just prior to the maximal growth of the organism and at a time when nutrients in the medium are exhausted; (ii) 4 days of incubation on a medium containing only inorganic salts and niacin or tryptophan; and (iii) subsequent illumination with light of wavelengths between 350 and 500 mμ. PMID:13883384

  17. A method for microbial decontamination ofAcanthamoeba cultures using the peritoneal cavity of mice

    Institute of Scientific and Technical Information of China (English)

    Rodrigo Gurgel-Gonalves; Patrcia Albuquerque; Csar Augusto Cuba-Cuba

    2015-01-01

    Objective:To evaluate whether the inoculation of contaminated cultures in the peritoneal cavity of mice could implement decontamination ofAcanthamoeba cultures. Methods: Suspensions ofAcanthamoeba,Acanthamoeba polyphagaATCC30461, or Acanthamoeba spp. isolated from soil (UnB13 strain) were inoculated in the peritoneal cavity of Swiss mice (n = 24). After 1, 6, 12, or 24 h of exposure the peritoneal cavity was washed and assessed for the presence of bacteria, fungi, andAcanthamoeba. Results: After 1 h of intraperitoneal inoculation at least 97% of the bacteria and 96% of the fungi (P Conclusions: Our data demonstrated that this technique has great potential for decontamination ofAcanthamoeba cultures in a short period of time.

  18. Analysis Of Usefulness Of Satellite Image Processing Methods For Investigations Of Cultural Heritage Resources

    Science.gov (United States)

    Osińska-Skotak, Katarzyna; Zapłata, Rafał

    2015-12-01

    The paper presents the analysis of usefulness of WorldView-2 satellite image processing, which enhance information concerning the cultural heritage objects. WorldView-2 images are characterised by the very high spatial resolution and high spectral resolution; that is why they create new possibilities for many applications, including investigations of the cultural heritage. The vicinities of Iłża have been selected as the test site for presented investigations. The presented results of works are the effect of research works, which were performed in the frames of the scientific project "Utilisation of laser scanning and remote sensing in protection, investigations and inventory of the cultural heritage. Development of non-invasive, digital methods of documenting and recognising the architectural and archaeological heritage", as the part of "The National Programme for the Development of Humanities" of the Minister of Science and Higher Education in the period of 2012-2015.

  19. Reflexive photography: an alternative method for documenting the learning process of cultural competence.

    Science.gov (United States)

    Amerson, Roxanne; Livingston, Wade G

    2014-04-01

    This qualitative descriptive study used reflexive photography to evaluate the learning process of cultural competence during an international service-learning project in Guatemala. Reflexive photography is an innovative qualitative research technique that examines participants' interactions with their environment through their personal reflections on images that they captured during their experience. A purposive sample of 10 baccalaureate nursing students traveled to Guatemala, where they conducted family and community assessments, engaged in home visits, and provided health education. Data collection involved over 100 photographs and a personal interview with each student. The themes developed from the photographs and interviews provided insight into the activities of an international experience that influence the cognitive, practical, and affective learning of cultural competence. Making home visits and teaching others from a different culture increased students' transcultural self-efficacy. Reflexive photography is a more robust method of self-reflection, especially for visual learners.

  20. In vitro culture of functionally active buffalo hepatocytes isolated by using a simplified manual perfusion method.

    Directory of Open Access Journals (Sweden)

    Santanu Panda

    Full Text Available In farm animals, there is no suitable cell line available to understand liver-specific functions. This has limited our understanding of liver function and metabolism in farm animals. Culturing and maintenance of functionally active hepatocytes is difficult, since they survive no more than few days. Establishing primary culture of hepatocytes can help in studying cellular metabolism, drug toxicity, hepatocyte specific gene function and regulation. Here we provide a simple in vitro method for isolation and short-term culture of functionally active buffalo hepatocytes.Buffalo hepatocytes were isolated from caudate lobes by using manual enzymatic perfusion and mechanical disruption of liver tissue. Hepatocyte yield was (5.3 ± 0.66×107 cells per gram of liver tissue with a viability of 82.3 ± 3.5%. Freshly isolated hepatocytes were spherical with well contrasted border. After 24 hours of seeding onto fibroblast feeder layer and different extracellular matrices like dry collagen, matrigel and sandwich collagen coated plates, hepatocytes formed confluent monolayer with frequent clusters. Cultured hepatocytes exhibited typical cuboidal and polygonal shape with restored cellular polarity. Cells expressed hepatocyte-specific marker genes or proteins like albumin, hepatocyte nuclear factor 4α, glucose-6-phosphatase, tyrosine aminotransferase, cytochromes, cytokeratin and α1-antitrypsin. Hepatocytes could be immunostained with anti-cytokeratins, anti-albumin and anti α1-antitrypsin antibodies. Abundant lipid droplets were detected in the cytosol of hepatocytes using oil red stain. In vitro cultured hepatocytes could be grown for five days and maintained for up to nine days on buffalo skin fibroblast feeder layer. Cultured hepatocytes were viable for functional studies.We developed a convenient and cost effective technique for hepatocytes isolation for short-term culture that exhibited morphological and functional characteristics of active hepatocytes

  1. The efficient method for simultaneous monitoring of the culturable as well as nonculturable airborne microorganisms.

    Science.gov (United States)

    Hubad, Barbara; Lapanje, Aleš

    2013-01-01

    Cultivation-based microbiological methods are a gold standard for monitoring of airborne micro-organisms to determine the occupational exposure levels or transmission paths of a particular infectious agent. Some highly contagious microorganisms are not easily culturable but it is becoming evident that cultivation and molecular methods are complementary and in these cases highly relevant. We report a simple and efficient method for sampling and analyzing airborne bacteria with an impactor-type high-flow-rate portable air sampler, currently used for monitoring culturable bacteria or fungi. A method is reported for extraction of nucleic acids from impacted cells without prior cultivation and using agarose as a sampling matrix. The DNA extraction efficiency was determined in spiked samples and, samples taken from a wastewater treatment plant and an alpine area. The abundance, diversity and quantity of total bacteria were analysed by a quantitative polymerase chain reaction, and by construction and analysis of clone libraries. The method does not interfere with downstream PCR analysis and can cover the gap between traditional culture and molecular techniques of bioaerosol monitoring.

  2. A simple and efficient method for the long-term preservation of plant cell suspension cultures

    Directory of Open Access Journals (Sweden)

    Boisson Anne-Marie

    2012-01-01

    Full Text Available Abstract Background The repeated weekly subculture of plant cell suspension is labour intensive and increases the risk of variation from parental cells lines. Most of the procedures to preserve cultures are based on controlled freezing/thawing and storage in liquid nitrogen. However, cells viability after unfreezing is uncertain. The long-term storage and regeneration of plant cell cultures remains a priority. Results Sycamore (Acer pseudoplatanus and Arabidopsis cell were preserved over six months as suspensions cultures in a phosphate-free nutrient medium at 5°C. The cell recovery monitored via gas exchange measurements and metabolic profiling using in vitro and in vivo 13C- and 31P-NMR took a couple of hours, and cell growth restarted without appreciable delay. No measurable cell death was observed. Conclusion We provide a simple method to preserve physiologically homogenous plant cell cultures without subculture over several months. The protocol based on the blockage of cell growth and low culture temperature is robust for heterotrophic and semi-autotrophic cells and should be adjustable to cell lines other than those utilised in this study. It requires no specialized equipment and is suitable for routine laboratory use.

  3. A method for microbial decontamination of Acanthamoeba cultures using the peritoneal cavity of mice简

    Institute of Scientific and Technical Information of China (English)

    Daniella; de; Sousa; Mendes; Moreira; Alves; Rodrigo; Gurgel-Gon?alves; Patrícia; Albuquerque; César; Augusto; Cuba-Cuba; Maria; Imaculada; Muniz-Junqueira; Selma; Aparecida; Souza; Kückelhaus

    2015-01-01

    Objective: To evaluate whether the inoculation of contaminated cultures in the peritoneal cavity of mice could implement decontamination of Acanthamoeba cultures.Methods: Suspensions of Acanthamoeba, Acanthamoeba polyphaga ATCC 30461, or Acanthamoeba spp. isolated from soil(Un B13 strain) were inoculated in the peritoneal cavity of Swiss mice(n = 24). After 1, 6, 12, or 24 h of exposure the peritoneal cavity was washed and assessed for the presence of bacteria, fungi, and Acanthamoeba.Results: After 1 h of intraperitoneal inoculation at least 97% of the bacteria and 96% of the fungi(P < 0.05) and 99% of the bacteria(P < 0.05) were successfully eliminated from the ATCC 30461 strain and from the soil isolate Un B13 strain, respectively. This method also allowed the recovery of most trophozoites and cysts from both Acanthamoeba cultures at the end of 24 h.Conclusions: Our data demonstrated that this technique has great potential for decontamination of Acanthamoeba cultures in a short period of time.

  4. Influence of the insemination method on the outcomes of elective blastocyst culture.

    Science.gov (United States)

    Wang, Caizhu; Feng, Guixue; Zhang, Bo; Shu, Jinhui; Zhou, Hong; Gan, Xianyou; Lin, Ruoyun

    2017-06-01

    The aim of this study was to explore the effects of the insemination method on the outcomes of elective blastocyst culture. We retrospectively analyzed the outcomes of elective blastocyst culture performed between January 2011 and December 2014. There were 2,003 cycles of conventional in vitro fertilization (IVF) and 336 cycles of intracytoplasmic sperm injection (ICSI), including 25,652 and 4,164 embryos that underwent sequential blastocyst culture, respectively. No significant differences were found in the female patients' age, basal follicle-stimulating hormone level, basal luteinizing hormone level, body mass index, number of oocytes, maturity rate, fertilization rate, or good-quality embryo rate. However, the blastocyst formation rate and embryo utilization rate were significantly higher in the conventional IVF group than in the ICSI group (54.70% vs. 50.94% and 51.09% vs. 47.65%, respectively, p0.05). No cycles were canceled due to the formation of no usable blastocysts. Although the fertilization method had no effect on clinical outcomes, the blastocyst formation rate and embryo utilization rate in the ICSI group were significantly lower than those observed in the conventional IVF group. Therefore, more care should be taken when choosing to perform blastocyst culture in ICSI patients.

  5. Human bone marrow cell culture: a sensitive method for determination of the biocompatibility of implant materials.

    Science.gov (United States)

    Wilke, A; Landgraff, M; Orth, J; Poenitz, H; Kienapfel, H; Boelte, K; Griss, P; Franke, R P

    1999-01-01

    The objective of this study was to develop a test method for determining the cytotoxicity and biocompatibility of various biomaterials that are used in orthopaedic surgery. This method is based on the use of a human bone marrow cell culture and was developed as an alternative to animal experiments. Human bone marrow cell culture has certain advantages over other cell culture models, as its results show a greater conformity with animal experimental results and clinical studies. Primary cell adherence, cell number, cell proliferation, production of extracellular matrix, cell viability and cell differentiation were used as indicative parameters of biocompatibility. After 2 weeks in culture, differences could be observed between the biomaterials with respect to these parameters. Cell numbers were greatest on the hydroxyapatite ceramic specimens, but were decreased on the titanium alloy specimens. Extracellular matrix hydroxyapatite production was high for ceramics, but reduced for titanium specimens. The polymers allowed only a few cells to adhere, and there were no signs of extracellular matrix production. The influence of biomaterials on differentiation of large numbers of cells was analysed by using flow cytophotometry. There were similar populations of T cells and monocytes on all specimens. However, extended B cell and granulocyte populations were observed with titanium and polyethylene.

  6. Using Educational Data Mining Methods to Assess Field-Dependent and Field-Independent Learners' Complex Problem Solving

    Science.gov (United States)

    Angeli, Charoula; Valanides, Nicos

    2013-01-01

    The present study investigated the problem-solving performance of 101 university students and their interactions with a computer modeling tool in order to solve a complex problem. Based on their performance on the hidden figures test, students were assigned to three groups of field-dependent (FD), field-mixed (FM), and field-independent (FI)…

  7. Simple screening method for molds producing intracellular mycotoxins in pure cultures.

    Science.gov (United States)

    Filtenborg, O; Frisvad, J C; Svendsen, J A

    1983-02-01

    A simple screening method for molds producing the intracellular mycotoxins brevianamide A, citreoviridin, cyclopiazonic acid, luteoskyrin, penitrem A, roquefortine C, sterigmatocystin, verruculogen, viomellein, and xanthomegnin was developed. After removing an agar plug from the mold culture, the mycelium on the plug is wetted with a drop of methanol-chloroform (1:2). By this treatment the intracellular mycotoxins are extracted within seconds and transferred directly to a thin-layer chromatography plate by immediately placing the plug on the plate while the mycelium is still wet. After removal of the plug, known thin-layer chromatographic procedures are carried out. The substrate (Czapek yeast autolysate agar) and growth conditions (25 degrees C for 7 days) used by Penicillium taxonomists proved suitable for the production of the mycotoxins investigated when 60 known toxigenic isolates and 865 cultures isolated from foods and feedstuffs were tested with this screening method.

  8. [Human bone marrow cell culture--a sensitive method for the evaluation of the biocompatibility of materials used in orthopedics].

    Science.gov (United States)

    Wilke, A; von Hirschheydt, S; Orth, J; Kienapfel, H; Griss, P; Franke, R P

    1995-01-01

    The objective of the study was to develop a test system to determine the cytotoxicity and biocompatibility of different biomaterials used in orthopedic surgery. This system was based on the use of a human bone marrow cell culture and the purpose was to find a screening method as a alternative to early animal experimental methods. The established human bone marrow cell culture has certain advantages when compared with other cell culture models. The result demonstrated a high conformity with animal experimental results.

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

    Directory of Open Access Journals (Sweden)

    Sutini Sutini

    2017-02-01

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

  10. The Containers Approach : An Inductive Method to look at Cultural Difference in a Technological Environment

    OpenAIRE

    Zimmermann, Basile

    2008-01-01

    The main goal of this presentation is to suggest an inductive way of looking at the idea of cultural difference in a technological environment. The proposal has three characteristics 1) it blurs the gap between natural sciences and human/social sciences, 2) it is extremely simple and down-to-earth, 3) the data that illustrates the method comes from recent field research in China (August 2008) in the realm of technology studies. The broader theoretical framework is provided by recent advances ...

  11. Survey on Heterotrophic Bacterial Contamination in Bottled Mineral Water by Culture Method

    OpenAIRE

    Essmaeel Ghorbanalinezhad; Ghazaleh Saeedi; Delaram Khanjani

    2014-01-01

    Background and Aim: This project focuses on the level of heterotrophic baceria in bottled mineral water which could be a health concern for the elderly, infants, pregnant women and immuno-compromised patients. Materials and Methods: Different brands of bottled water samples were selected randomly and evaluated for their bacteriological quality, using different specific culture media and biochemical tests. Water samples were analyzed within 24 hours of their purchase/collection. Samples we...

  12. Prevalence of periodontopathogenic bacteria in patients suffering from periodontitis using culture and PCR methods

    Directory of Open Access Journals (Sweden)

    Amir Aliramezani

    2012-01-01

    Full Text Available Background and Aims: Periodontitis is one of the most common oral diseases with the various incidence rates in different populations. A number of bacteria are considered as the major etiologic agents of periodontitis. The aim of the present study was to determine the prevalence of periodontopathogen bacteria in patients using both PCR and culture techniques.Materials and Methods: In this study, one-hundred patients (including 62 females and 38 males with an average age of 49±11.5 years with adult periodontitis referred to periodontics department of School of Dentistry/Tehran University of Medical Sciences were investigated. The samples were taken and sent immediately to the laboratory for culture and molecular evaluation. The PCR was performed using specific primers and the statistical analysis of data was performed using SPSS statistic software and McNemar test.Results: The results demonstrated that the total detection rate in culture method was 64%. The rate of Aggregatibacter actinomycetemcomitans (Aa was 28% which was significantly higher than that of Porphyromonas gingivalis (Pg (6% and Prevotella intermedia (Pi (3%. 27% of cases showed mixed bacterial growth. 65% of patients were positive using molecular method. The rate of Aa (30% was significantly higher than that of Pg (7% and Pi (5%. The mixed PCR positive rate containing of Aa, Pg and Pi was (23%.Conclusion: In this study, it was found that most of the bacteria isolated using culture and molecular methods were Aa, Pg and Pi, respectively. Although the detection frequencies of both techniques were similar, the specificity, sensitivity and bacterial detection speed of the PCR technique is obviously higher. Therefore, the use of molecular techniques is strongly recommended. However, both techniques seem to be suitable for microbiological diagnostics.

  13. Culturally Authentic Scaling Approach: A Multi-Step Method for Culturally Adapting Measures for Use with Ethnic Minority and Immigrant Youths

    Directory of Open Access Journals (Sweden)

    Guerda Nicolas

    2009-03-01

    Full Text Available Given the increasing ethnic and racial diversity of youths in the U.S., researchers must be conscious of how youth are being recruited, retained, and assessed in research programs. In this article, we describe an efficient and replicable methodology, the Culturally Authentic Scaling Approach (CASA, which can be implemented to culturally adapt measures for use with ethnic minority and immigrant youths. Specifically, the steps involved in the CASA method are described, including developing community partnership, evaluating the theoretical equivalence, adjusting the selection and administration of measures. Engaging in an on-going dialogue with the community to increase cultural validity and build community relationships is also discussed. Addressing the cultural validity of measures used with ethnic and immigrant youths enhances the probability that the information obtained will be reflective of the cultural background of the participants and an accurate assessment of their experiences

  14. Ethnonursing: A Qualitative Research Method for Studying Culturally Competent Care across Disciplines

    Directory of Open Access Journals (Sweden)

    Marilyn R. McFarland PhD, RN, FNP-BC, CTN

    2012-07-01

    Full Text Available Nurse anthropologist, Madeleine Leininger, developed the culture care theory and ethnonursing research method to help researchers study transcultural human care phenomena and discover the knowledge nurses need to provide care in an increasingly multicultural world. The authors propose that the ethnonursing method can be useful for research that addresses providing care in other disciplines, including education, administration, physical, occupational, and speech therapy, social work, pharmacy, medicine, and other disciplines in which research findings have implications for human care and health. The authors discuss the culture care theory and describe the ethnonursing research method's enablers, data analysis phases, and qualitative evaluation criteria. The theory is presented as a guide for using research findings to design culturally competent and congruent care to promote well-being among diverse people, groups, communities, and institutions. Resources include a reference list of key source publications, a discussion of exemplar studies, and samples of a theory-based, open-ended interview guide and data coding system.

  15. The State of the Art of Teaching Research Methods in the Social Sciences: Towards a Pedagogical Culture

    Science.gov (United States)

    Wagner, Claire; Garner, Mark; Kawulich, Barbara

    2011-01-01

    No formal pedagogical culture for research methods in the social sciences seems to exist and, as part of the authors' endeavour to establish such a culture, this article reviews current literature about teaching research methods and identifies the gaps in the research. Articles in academic journals spanning a 10-year period were collected by…

  16. The State of the Art of Teaching Research Methods in the Social Sciences: Towards a Pedagogical Culture

    Science.gov (United States)

    Wagner, Claire; Garner, Mark; Kawulich, Barbara

    2011-01-01

    No formal pedagogical culture for research methods in the social sciences seems to exist and, as part of the authors' endeavour to establish such a culture, this article reviews current literature about teaching research methods and identifies the gaps in the research. Articles in academic journals spanning a 10-year period were collected by…

  17. MHC multimer-guided and cell culture-independent isolation of functional T cell receptors from single cells facilitates TCR identification for immunotherapy.

    Directory of Open Access Journals (Sweden)

    Georg Dössinger

    Full Text Available Adoptive therapy using T cells redirected to target tumor- or infection-associated antigens is a promising strategy that has curative potential and broad applicability. In order to accelerate the screening process for suitable antigen-specific T cell receptors (TCRs, we developed a new approach circumventing conventional in vitro expansion-based strategies. Direct isolation of paired full-length TCR sequences from non-expanded antigen-specific T cells was achieved by the establishment of a highly sensitive PCR-based T cell receptor single cell analysis method (TCR-SCAN. Using MHC multimer-labeled and single cell-sorted HCMV-specific T cells we demonstrate a high efficacy (approximately 25% and target specificity of TCR-SCAN receptor identification. In combination with MHC-multimer based pre-enrichment steps, we were able to isolate TCRs specific for the oncogenes Her2/neu and WT1 even from very small populations (original precursor frequencies of down to 0.00005% of CD3(+ T cells without any cell culture step involved. Genetic re-expression of isolated receptors demonstrates their functionality and target specificity. We believe that this new strategy of TCR identification may provide broad access to specific TCRs for therapeutically relevant T cell epitopes.

  18. Differentiation of Salmonella enterica serovars and strains in cultures and food using infrared spectroscopic and microspectroscopic techniques combined with soft independent modeling of class analogy pattern recognition analysis.

    Science.gov (United States)

    Männig, Annegret; Baldauf, Nathan A; Rodriguez-Romo, Luis A; Yousef, Ahmed E; Rodríguez-Saona, Luis E

    2008-11-01

    Detection of pathogenic microorganisms in food is often a tedious and time-consuming exercise. Developing rapid and cost-effective techniques for identifying pathogens to subspecies is critical for tracking causes of foodborne disease outbreaks. The objective of this study was to develop a method for rapid identification and differentiation of Salmonella serovars and strains within these serovars through isolation on hydrophobic grid membrane filters (HGMFs), examination by infrared (IR) spectroscopy and microspectroscopy, and data analysis by multivariate statistical techniques. Salmonella serovars (Anatum, Enteritidis, Heidelberg, Kentucky, Muenchen, and Typhimurium), most of which were represented by multiple strains, were grown in tryptic soy broth (24 h at 42 degrees C), diluted to 10(2) to 10(3) CFU/ml, and filtered using HGMFs. The membranes were incubated on Miller-Mallinson agar (24 h at 42 degrees C), and typical Salmonella colonies were sonicated in 50% acetonitrile and centrifuged. Resulting pellets were vacuum dried on a ZnSe crystal and analyzed using IR spectroscopy. Alternatively, the membranes containing Salmonella growth were removed from the agar, vacuum dried, and colonies were analyzed directly by IR microspectroscopy. Soft independent modeling of class analogy (SIMCA) models were developed from spectra. The method was validated by analyzing Salmonella-inoculated tomato juice, eggs, milk, and chicken. Salmonella serovars exhibited distinctive and reproducible spectra in the fingerprint region (1,200 to 900 cm(-1)) of the IR spectrum. SIMCA permitted distinguishing Salmonella strains from each other through differences in bacterial lipopolysaccharides and other membrane components. The model correctly predicted Salmonella in foods at serovar (100%) and strain (90%) levels. Isolation of Salmonella on HGMF and selective agar followed by IR spectroscopic analysis resulted in rapid and efficient isolation, identification, and differentiation of

  19. Critical assessment of extracellular polymeric substances extraction methods from mixed culture biomass

    DEFF Research Database (Denmark)

    Pellicer i Nàcher, Carles; Domingo Felez, Carlos; Mutlu, Ayten Gizem

    2013-01-01

    . This study presents a rigorous and critical assessment of existing physical and chemical EPS extraction methods applied to mixed-culture biomass samples (nitrifying, nitritation-anammox, and activated sludge biomass). A novel fluorescence-based method was developed and calibrated to quantify the lysis...... potential of different EPS extraction protocols. We concluded that commonly used methods to assess cell lysis (DNA concentrations or G6PDH activities in EPS extracts) do not correlate with cell viability. Furthermore, we discovered that the presence of certain chemicals in EPS extracts results in severe...... underestimation of protein and carbohydrate concentrations by using standard analytical methods. Keeping both maximum EPS extraction yields and minimal biomass lysis as criteria, it was identified a sonication-based extraction method as the best to determine and compare tightly-bound EPS fractions in different...

  20. Co-interviewing across gender and culture: expanding qualitative research methods in Melanesia.

    Science.gov (United States)

    Redman-MacLaren, Michelle L; Api, Unia K; Darius, Matupit; Tommbe, Rachael; Mafile'o, Tracie A; MacLaren, David J

    2014-09-06

    The social and cultural positions of both researchers and research participants influence qualitative methods and study findings. In Papua New Guinea (PNG), as in other contexts, gender is a key organising characteristic and needs to be central to the design and conduct of research. The colonial history between researcher and participant is also critical to understanding potential power differences. This is particularly relevant to public health research, much of which has emerged from a positivist paradigm. This paper describes our critical reflection of flexible researcher responses enacted during qualitative research in PNG. Led by a senior male HIV researcher from PNG, a male from a PNG university and a female from an Australian university conducted qualitative interviews about faith-based responses to HIV in PNG. The two researchers planned to conduct one-on-one interviews matching gender of participants and interviewer. However, while conducting the study, four participants explicitly requested to be interviewed by both researchers. This experience led us to critically consider socially and culturally situated ways of understanding semi-structured interviewing for public health research in Melanesia. New understandings about public health research include: (i) a challenge to the convention that the researcher holds more power than the research participant, (ii) the importance of audience in Melanesia, (iii) cultural safety can be provided when two people co-interview and (iv) the effect an esteemed leader heading the research may have on people's willingness to participate. Researchers who occupy insider-outsider roles in PNG may provide participants new possibilities to communicate key ideas. Our recent experience has taught us public health research methods that are gender sensitive and culturally situated are pivotal to successful research in Melanesia. Qualitative research requires adaptability and reflexivity. Public health research methods must continue

  1. A simple micro-culture method for the study of group B arboviruses*

    Science.gov (United States)

    de Madrid, Ana Teresa; Porterfield, James S.

    1969-01-01

    Thirty-nine group B arboviruses have been titrated by a simple micro-culture method. The technique uses a stable line of pig kidney cells (PS cells) in which plaques develop when cells are first infected in suspension in the wells of haemagglutination trays and are then incubated for from 3 to 10 days under an overlay containing carboxymethyl-cellulose. This method can be adapted to measure neutralizing antibodies, and the principle underlying the test is applicable to other cells and other viruses. ImagesFIG. 1FIG. 2FIG. 3FIG. 4 PMID:4183812

  2. Different Donor Cell Culture Methods Can Influence the Developmental Ability of Cloned Sheep Embryos.

    Directory of Open Access Journals (Sweden)

    LiBing Ma

    Full Text Available It was proposed that arresting nuclear donor cells in G0/G1 phase facilitates the development of embryos that are derived from somatic cell nuclear transfer (SCNT. Full confluency or serum starvation is commonly used to arrest in vitro cultured somatic cells in G0/G1 phase. However, it is controversial as to whether these two methods have the same efficiency in arresting somatic cells in G0/G1 phase. Moreover, it is unclear whether the cloned embryos have comparable developmental ability after somatic cells are subjected to one of these methods and then used as nuclear donors in SCNT. In the present study, in vitro cultured sheep skin fibroblasts were divided into four groups: (1 cultured to 70-80% confluency (control group, (2 cultured to full confluency, (3 starved in low serum medium for 4 d, or (4 cultured to full confluency and then further starved for 4 d. Flow cytometry was used to assay the percentage of fibroblasts in G0/G1 phase, and cell counting was used to assay the viability of the fibroblasts. Then, real-time reverse transcription PCR was used to determine the levels of expression of several cell cycle-related genes. Subsequently, the four groups of fibroblasts were separately used as nuclear donors in SCNT, and the developmental ability and the quality of the cloned embryos were compared. The results showed that the percentage of fibroblasts in G0/G1 phase, the viability of fibroblasts, and the expression levels of cell cycle-related genes was different among the four groups of fibroblasts. Moreover, the quality of the cloned embryos was comparable after these four groups of fibroblasts were separately used as nuclear donors in SCNT. However, cloned embryos derived from fibroblasts that were cultured to full confluency combined with serum starvation had the highest developmental ability. The results of the present study indicate that there are synergistic effects of full confluency and serum starvation on arresting fibroblasts in

  3. Comparative and multilaboratory studies of two immunodiffusion method enrichment protocols and the AOAC/Bacteriological Analytical Manual culture method for detection of Salmonella in all foods.

    Science.gov (United States)

    Feldsine, P T; Falbo-Nelson, M T; Hustead, D L; Flowers, R S; Flowers, M J

    1995-01-01

    The single enrichment immunodiffusion (1-step), the preenrichment and selective enrichment immunodiffusion (2-step), and the AOAC/Bacteriological Analytical Manual culture methods for Salmonella were evaluated for equivalence in 2 separate studies, a comparative evaluation and a multilaboratory dilution study. In the comparative study, all 3 methods were performed on 10 food types. For 550 samples, analyses resulted in 99.3 and 99.6% agreement between the culture method and the 1-step and 2-step methods, respectively. False negative rates were 0.9 and 0.3% for 1-step and culture, and 0.0% and 0.6% for the 2-step and culture, respectively. Subsequently, 6 food types were included in a multilaboratory dilution-to-extinction study. A sequential dilution series of Salmonella in foods was analyzed by the 3 methods to determine their lower limits of detection for Salmonella. A total of 1185 samples analyzed resulted in 98.9% agreement between 1-step and culture, and 99.7% agreement between 2-step and culture. False negative rates were 1.8 and 0.1% for 1-step and culture, and 0.4 and 0.1% for 2-step and culture, respectively. During these evaluations, 1735 samples and controls representing 10 different naturally contaminated and inoculated foods were tested. The data indicate statistical equivalence of all 3 methods when analyzing all food types.

  4. Explant culture: An advantageous method for isolation of mesenchymal stem cells from human tissues.

    Science.gov (United States)

    Hendijani, Fatemeh

    2017-04-01

    Mesenchymal stem cell (MSC) research progressively moves towards clinical phases. Accordingly, a wide range of different procedures were presented in the literature for MSC isolation from human tissues; however, there is not yet any close focus on the details to offer precise information for best method selection. Choosing a proper isolation method is a critical step in obtaining cells with optimal quality and yield in companion with clinical and economical considerations. In this concern, current review widely discusses advantages of omitting proteolysis step in isolation process and presence of tissue pieces in primary culture of MSCs, including removal of lytic stress on cells, reduction of in vivo to in vitro transition stress for migrated/isolated cells, reduction of price, processing time and labour, removal of viral contamination risk, and addition of supporting functions of extracellular matrix and released growth factors from tissue explant. In next sections, it provides an overall report of technical highlights and molecular events of explant culture method for isolation of MSCs from human tissues including adipose tissue, bone marrow, dental pulp, hair follicle, cornea, umbilical cord and placenta. Focusing on informative collection of molecular and methodological data about explant methods can make it easy for researchers to choose an optimal method for their experiments/clinical studies and also stimulate them to investigate and optimize more efficient procedures according to clinical and economical benefits. © 2017 John Wiley & Sons Ltd.

  5. Heat-transfer-method-based cell culture quality assay through cell detection by surface imprinted polymers.

    Science.gov (United States)

    Eersels, Kasper; van Grinsven, Bart; Khorshid, Mehran; Somers, Veerle; Püttmann, Christiane; Stein, Christoph; Barth, Stefan; Diliën, Hanne; Bos, Gerard M J; Germeraad, Wilfred T V; Cleij, Thomas J; Thoelen, Ronald; De Ceuninck, Ward; Wagner, Patrick

    2015-02-17

    Previous work has indicated that surface imprinted polymers (SIPs) allow for highly specific cell detection through macromolecular cell imprints. The combination of SIPs with a heat-transfer-based read-out technique has led to the development of a selective, label-free, low-cost, and user-friendly cell detection assay. In this study, the breast cancer cell line ZR-75-1 is used to assess the potential of the platform for monitoring the quality of a cell culture in time. For this purpose, we show that the proposed methodology is able to discriminate between the original cell line (adherent growth, ZR-75-1a) and a descendant cell line (suspension growth, ZR-75-1s). Moreover, ZR-75-1a cells were cultured for a prolonged period of time and analyzed using the heat-transfer method (HTM) at regular time intervals. The results of these experiments demonstrate that the thermal resistance (Rth) signal decays after a certain number of cell culture passages. This can likely be attributed to a compromised quality of the cell culture due to cross-contamination with the ZR-75-1s cell line, a finding that was confirmed by classical STR DNA profiling. The cells do not express the same functional groups on their membrane, resulting in a weaker bond between cell and imprint, enabling cell removal by mechanical friction, provided by flushing the measuring chamber with buffer solution. These findings were further confirmed by HTM and illustrate that the biomimetic sensor platform can be used as an assay for monitoring the quality of cell cultures in time.

  6. Development of a Novel Nuclear Safety Culture Evaluation Method for an Operating Team Using Probabilistic Safety Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Han, Sangmin; Lee, Seung Min; Seong, Poong Hyun [KAIST, Daejeon (Korea, Republic of)

    2015-05-15

    IAEA defined safety culture as follows: 'Safety Culture is that assembly of characteristics and attitudes in organizations and individuals which establishes that, as an overriding priority, nuclear plant safety issues receive the attention warranted by their significance'. Also, celebrated behavioral scientist, Cooper, defined safety culture as,'safety culture is that observable degree of effort by which all organizational members direct their attention and actions toward improving safety on a daily basis' with his internal psychological, situational, and behavioral context model. With these various definitions and criteria of safety culture, several safety culture assessment methods have been developed to improve and manage safety culture. To develop a new quantitative safety culture evaluation method for an operating team, we unified and redefined safety culture assessment items. Then we modeled a new safety culture evaluation by adopting level 1 PSA concept. Finally, we suggested the criteria to obtain nominal success probabilities of assessment items by using 'operational definition'. To validate the suggested evaluation method, we analyzed the collected audio-visual recording data collected from a full scope main control room simulator of a NPP in Korea.

  7. Gulliver's Eggs: Why Methods are not an Issue of Qualitative Research in Cultural Psychology.

    Science.gov (United States)

    Tateo, Luca

    2015-06-01

    The future of qualitative methods regards the kind of object cultural psychology is interested and the kind of questions it can ask. I propose that the object should be experiencing, understood as a complex whole, consisting of lived-by action and counter-action, that is contextual inter-action with the world in the form of an experiencing subject and otherness. The kind of questions cultural psychology can ask is instead related to the epistemological status attributed to both researcher and participant. Probably few scholars such as Vygotsky, Piaget and Lewin understood to what extent experiencing is always changing, because the relationship between mind, alterity and culture is co-generative. This also implies a relativization and a decentralization of the psychology's perspective. Finally, I provide some examples from the history of psychology and some suggestions to work at the level of such complexity by using methods that can work with complex objects such as products of human activity (e.g., art, literature, architecture, etc.).

  8. A machine-independent method to have active removal of 5,000 centistokes silicone oil using plastic infusion tube and 23-gauge microcannulas

    OpenAIRE

    Zhang, Zhaotian; Wei, Yantao; Jiang, Xintong; Qiu, Suo; Zhang, Shaochong

    2015-01-01

    Background To describe one modified method of having machine-independent removal of 5,000 centistokes silicone oil through 23-gauge trocar-cannulas. Methods Consecutive patients with silicone oil tamponade for more than four months and with complete retinal reattachment were included. Two 23-gauge trocars were used to make sclerotomies while the microcannulas remained in situ for intravitreous infusion and silicone oil drainage. A short section of infusion tube was connected with a 10 ml syri...

  9. Cell culture methods for the establishment of the NCI series of lung cancer cell lines.

    Science.gov (United States)

    Oie, H K; Russell, E K; Carney, D N; Gazdar, A F

    1996-01-01

    More than 200 human small cell lung cancer and non-small cell lung cancer cell lines were established over 15 years mainly by utilizing the serum-free, hormone and growth factor supplemented, defined media HITES and ACL4. Use of modified, established cell culture techniques such as the mechanical spillout method for the releasing of cell aggregates from tumor tissue, ficoll gradient centrifugation for the separation of tumor cells from erythrocytes and tissue debris, and an apparatue consisting of a platinum tubing attached to a suction flask for removal of spent medium have greatly contributed to the success in culturing tumor cells. Characterization of these lung cancer cell lines have extended our knowledge of lung cell biology. Studies elucidating the nutritional requirements of lung cancer cell growth may be helpful for the manipulation of these tumors in patients.

  10. A Culture Method for Darkling Beetles, Blapstinus spp. (Coleoptera:Tenebrionidae).

    Science.gov (United States)

    Zilkowski, Bruce W; Cossé, Allard A

    2015-06-01

    Darkling beetles, Blapstinus spp., have become a serious pest of Cucurbitaceae crops, especially in California. A culture method was sought to provide large numbers (>500) of adult beetles of known age and sex that could be used for laboratory testing when needed. A method previously developed for Alphitobius diaperinus (Panzer) using a diet of ground chick feed, with apple slices as a moisture source, was modified for use with Blapstinus spp. and then compared with the same method substituting apple slices with zucchini as the moisture source. Rearing boxes set up with apple slices produced significantly more pupae and adults than boxes containing zucchini slices. However, using either zucchini or apples as a moisture source yielded over the target of 500 adults per rearing box. A previous method designed to sex A. diaperinus based on the presence (♀) or absence (♂) of second valvifers in the pupal stage also proved to be effective for sexing the Blapstinus spp.

  11. Characterization by culture-dependent and culture-1 independent methods of the 2 bacterial population of suckling-lamb packaged in different atmospheres

    NARCIS (Netherlands)

    Oses, S.M.; Diez, A.M.; Melero, B.; Luning, P.A.; Jaime, I.; Rovira, J.

    2013-01-01

    This study offers insight into the dynamics of bacterial populations in fresh cuts of suckling lamb under four different atmospheric conditions: air (A), and three Modified Atmosphere Packaging (MAP) environments, 15%O2/30%CO2/55%N2 (C, commercial), 70%O2/30%CO2 (O), and 15%O2/85%CO2 (H) for 18

  12. Characterization by culture-dependent and culture-1 independent methods of the 2 bacterial population of suckling-lamb packaged in different atmospheres

    NARCIS (Netherlands)

    Oses, S.M.; Diez, A.M.; Melero, B.; Luning, P.A.; Jaime, I.; Rovira, J.

    2013-01-01

    This study offers insight into the dynamics of bacterial populations in fresh cuts of suckling lamb under four different atmospheric conditions: air (A), and three Modified Atmosphere Packaging (MAP) environments, 15%O2/30%CO2/55%N2 (C, commercial), 70%O2/30%CO2 (O), and 15%O2/85%CO2 (H) for 18 days

  13. Analysis of cultural development of Isfahan city Using Factor analysis method

    Directory of Open Access Journals (Sweden)

    J.Mohammadi

    2013-01-01

    Full Text Available Extended abstract1-IntroductionCultural spaces are consideredas one of the main factors for development. Cultural development is a qualitative and valuable process that for assessing it, quantitative indicators in cultural planning are used to obtain development objectives in the pattern of goods and services. The aim of the study is to determine and analyze cultural development level and regional inequality of different districts of Isfahan using factor analysis technique. The statistical population of the study is 14 districts of Isfahan municipality. The dominant approach ofthis study is quantitative – description and analytical. In this study, 35 indices have been summarized by factor analysis method and have been reduced to 5 factors and combined in significant ones and delivered.2 – Theoretical basesThe most important objectives of spatial planning, considering limitation of resources, are optimum distributions of facilities and services among different locations in which people live. To do this,there is a need to identify different locations in terms of having different facilities and services, so that developed locations are specified and planners can proceed to do something for spatial equilibrium and reducing privileged distance between districts.The present study has been conducted to reach to an equal development in Isfahan urban districts following identifying the situation and the manner of distributing development facilities cultural selected indices in different districts.3 – DiscussionCultural development of societies is evaluated by considering the changes and improvement of its indices and measured by quantitative frames. Cultural development indices are the most important tools for cultural planning in a special district in a society. In this study, cultural development indices have been used to determine the levels of districts. By using factor analysis model, the share of influential factors in the cultural

  14. Organizational culture change in U.S. hospitals: a mixed methods longitudinal intervention study.

    Science.gov (United States)

    Curry, Leslie A; Linnander, Erika L; Brewster, Amanda L; Ting, Henry; Krumholz, Harlan M; Bradley, Elizabeth H

    2015-03-07

    Improving outcomes for patients with acute myocardial infarction (AMI) is a priority for hospital leadership, clinicians, and policymakers. Evidence suggests links between hospital organizational culture and hospital performance; however, few studies have attempted to shift organizational culture in order to improve performance, fewer have focused on patient outcomes, and none have addressed mortality for patients with AMI. We sought to address this gap through a novel longitudinal intervention study, Leadership Saves Lives (LSL). This manuscript describes the methodology of LSL, a 2-year intervention study using a concurrent mixed methods design, guided by open systems theory and the Assess, Innovate, Develop, Engage, Devolve (AIDED) model of diffusion, implemented in 10 U.S. hospitals and their peer hospital networks. The intervention has three primary components: 1) annual convenings of the ten intervention hospitals; 2) semiannual workshops with guiding coalitions at each hospital; and 3) continuous remote support across all intervention hospitals through a web-based platform. Primary outcomes include 1) shifts in key dimensions of hospital organizational culture associated with lower mortality rates for patients with AMI; 2) use of targeted evidence-based practices associated with lower mortality rates for patients with AMI; and 3) in-hospital AMI mortality. Quantitative data include annual surveys of guiding coalition members in the intervention hospitals and peer network hospitals. Qualitative data include in-person, in-depth interviews with all guiding coalition members and selective observations of key interactions in care for patients with AMI, collected at three time points. Data integration will identify patterns and major themes in change processes across all intervention hospitals over time. LSL is novel in its use of a longitudinal mixed methods approach in a diverse sample of hospitals, its focus on objective outcome measures of mortality, and its

  15. [Preliminary Study of Lonicera hypoglauca on Germination Conditions of Sand Culture Seeds and Sterilization Method of Sand Culture Seedling Sterilization].

    Science.gov (United States)

    Tan, Mu-xiu; Zeng, Wen-wen; Wei, Peng-xiao; Mo, Qiao-cheng; Pu, Zu-ning; Cen, Xiu-fen; Shi, Feng-hua

    2015-05-01

    To explore the germination conditions of Lonicera hypoglauca sand culture seeds and the effects of sand culture seedlings sterilization. 0.1% HgCl2 with different sterilization time, different illumination time and temperature culture condition were adopted to study the germination conditions of sand culture seeds. Different sterilization treatments and different hardening-seedling days were used to test the sterilization effect of sand culture seedlings. The sterilization effect of the combination of 75% ethanol 30 s + 0.1% HgCl2 5 min on Lonicera hypoglauca seeds was the optimum,with the average pollution rate of 15.56%, and the average germination rate reached 51.11%. The combination of varied temperature-room temperature under light for 12 h/d was the best, with the average germination rate peaked at 75.49%, and the average germination potential reached 68.36%. The treatment of detergent liquor scrub-tap water wash on the part above the hypocotyl, which was sand cultured under the opening condition and had no root, showed the best sterilization effect, with the average pollution rate was zero, and the average survival rate peaked at 100.00%. The sterilization effect of sand culture seedlings, which was disinfected after cleaning by detergent liquor scrub-tap water wash after hardening-seeding for 30 days, was the best, with the average pollution rate of 50.00%, and the average survival rate of 100.00%. The best sterilization effect is the combination of 75% ethanol 30 s + 0.1% HgCl2 5 min; Lighting for 12 h/d of varied temperature-room temperature is regarded as the optimum culture condition. The treatment of detergent liquor scrub-tap water wash treatment on the part above the hypocotyl,which is sand cultured under the opening condition and had no root, shows the best sterilization effect. For the sand culture seedlings, before inoculated in subculture medium, should be hardening-seedling for some days and sterilized after detergent liquor scrub-tap water wash.

  16. A new method for extracting the ENSO-independent Indian Ocean Dipole: application to Australian region tropical cyclone counts

    Energy Technology Data Exchange (ETDEWEB)

    Werner, Angelika; Maharaj, Angela M. [Macquarie University, Department of Environment and Geography, Sydney, NSW (Australia); Holbrook, Neil J. [University of Tasmania, School of Geography and Environmental Studies, Hobart, TAS (Australia)

    2012-06-15

    We introduce a simple but effective means of removing ENSO-related variations from the Indian Ocean Dipole (IOD) in order to better evaluate the ENSO-independent IOD contribution to Australian climate - specifically here interannual variations in Australian region tropical cyclogensis (TCG) counts. The ENSO time contribution is removed from the Indian Ocean Dipole Mode index (DMI) by first calculating the lagged regression of the DMI on the sea surface temperature anomaly (SSTA) index NINO3.4 to maximum lags of 8 months, and then removing this ENSO portion. The new ENSO-independent time series, DMI{sub NOENSO}, correlates strongly with the original DMI at r = 0.87 (significant at >99% level). Despite the strength of the correlation between these series, the IOD events classified based on DMI{sub NOENSO} provide important differences from previously identified IOD events, which are more closely aligned with ENSO phases. IOD event composite maps of SSTAs regressed on DMI{sub NOENSO} reveal a much greater ENSO-independence than the original DMI-related SSTA pattern. This approach is used to explore relationships between Australian region TCG and IOD from 1968 to 2007. While we show that both the DMI and DMI{sub NOENSO} have significant hindcast skill (on the 95% level) when used as predictors in a multiple linear regression model for Australian region annual TCG counts, the IOD does not add any significant hindcast skill over an ENSO-only predictor model, based on NINO4. Correlations between the time series of annual TCG count observations and ENSO + IOD model cross-validated hindcasts achieve r = 0.68 (significant at the 99% level). (orig.)

  17. Evaluation of methods to prepare samples of leafy green vegetables for preenrichment with the Bacteriological Analytical Manual Salmonella culture method.

    Science.gov (United States)

    Jacobson, Andrew Patrick; Gill, Vikas Singh; Irvin, Kari Anne; Wang, Hua; Hammack, Thomas Seyliard

    2012-02-01

    Three sample preparation procedures, soak, stomach, and blend, were evaluated using the Bacteriological Analytical Manual Salmonella culture method with eight types of leafy green produce. In the soak method, test portions were added to lactose broth without homogenization; in the stomach method, test portions were stomached with lactose broth; and in the blend method, test portions were blended with lactose broth. Twenty artificially contaminated test portions were analyzed with each procedure in individual experimental trials. The number of test portions identified as positive were compared among the procedures. Statistically significant differences were identified with Fisher's exact two-tailed F test (P < 0.05). Where differences did occur (P < 0.05), the soak procedure was the most effective or was at least as effective as homogenization by either blending or stomaching. Statistically significant differences most frequently occurred with romaine lettuce and cabbage; for these items, blending was significantly less effective than the soak procedure. Overall, for all of the produce types examined, results showed that the soak procedure was more effective than either of the homogenization procedures in recovering Salmonella from leafy green produce. Of the 540 test portions examined by each sample preparation method, 344 were positive for the presence of Salmonella by soaking, 293 by stomaching, and 232 by blending. We recommend that the soak procedure replace homogenization for the analysis of leafy green produce because the soak procedure is more productive than homogenization by either blending or stomaching of the leafy green produce types as reported herein.

  18. [Comparison of direct microscopy, culture, ELISA and molecular methods for diagnosis of Entamoeba histolytica].

    Science.gov (United States)

    Tüzemen, Nazmiye Ulkü; Doğan, Nihal

    2014-01-01

    Amebiasis, a parasitic infection caused by Entamoeba histolytica, is one of the most common parasitic infections worldwide. Since it is still an important public health problem in developing countries, rapid differential diagnosis of amebiasis is crucial in terms of treatment. The most frequently used method for laboratory diagnosis is direct microscopy, however more reliable and specific methods are needed in order to differentiate the apathogenic Entamoeba dispar under the microscope. This study was conducted to compare the results of different methods namely, direct microscopy, culture, ELISA and PCR for the detection of E.histolytica in stool samples and to evaluate the performances of those methods. A total of 1049 stool samples collected from pediatric and adult patients who were admitted to hospital with diarrhea complaint between January 2011-March 2013, and randomly selected samples from primary school children, were included in the study. Direct microscopic examination was performed by native-lugol, physiological saline, modified formol-ethyl acetate sedimentation and trichrome staining methods. The stool samples were also inoculated into TYI-S-33 media for axenic cultivation of amoeba. The presence of amebic antigens in the samples were screened by a commercial ELISA kit (TechLab, E.histolytica II, USA). For the molecular diagnosis, a multiplex tandem real-time PCR (MT-PCR) kit (AusDiagnostics Pty Ltd, Australia) was used, after the extraction of DNAs with QIAamp DNA Stool Mini Kit (Qiagen, USA). A total of 354 samples which could be evaluated by all of the methods, were included in the study. Of the 354 stool samples, 84 (23.7%) were found E.histolytica/E.dispar positive by direct microscopy, 61 (17.2%) by trichrome staining, 46 (12.9%) by culture, 31 (8.7%) by ELISA and 9 (2.5%) by MT-PCR. Of direct microscopy positive samples 54.7% (46/84) were also positive with trichrome staining, 39.3% (33/84) with culture, 15.5% (13/84) with ELISA and 7.1% (6

  19. Oral rinse as a potential method to culture Candida isolate from AIDS patients

    Directory of Open Access Journals (Sweden)

    Desiana Radithia

    2011-12-01

    Full Text Available Background: Candida isolate is easily sampled from oral cavity by swabbing directly on the candidiasis lesion, to be smeared onto slides for direct examination or cultured in a growth medium. This method is by far the gold standard for defining candidiasis diagnosis. However it is difficult to apply on sensitive patients and almost impossible on patients showing no clinical appearance of oral candidiasis. AIDS patients are very prone to candida infection and have a tendency of repetitive infection involving mixed species. As many candida species show different susceptibility to anti-fungal agents, it is necessary to identify the species causing the infection in the management of oral candidiasis. Oral rinse is a suggested method to obtain candida isolate to be cultured for further analysis such as species identification. This method is simple and less risky on infection transmission as less tools are required in the procedure. Purpose: This study aimed to assess the application of oral rinse as an alternative method to culture Candida isolate from AIDS patients. Methods: A cross-sectional observative study was conducted in HIV/AIDS in-Patient Facility of Intermediate Care Unit for Infection Disease, Dr. Soetomo Hospital Surabaya. Fourteen stadium 4 AIDS patients matching criteria were swabbed on 1/3-posterior of the tongue, and then given 10 ml phosphate buffer saline to rinse vigorously for 15 seconds. Both specimens were cultured on Sabouraud’s dextrose agar and colony growth was observed. Results: Candida colonies were able to grow from all 14 isolates (100% by both methods. Qualitatively, cultures from oral rinse specimens were more populated than cultures from swab specimens. Conclusion: Oral rinse is an applicable technique to obtain Candida species isolate. This technique is safe, easy, non-invasive, and needs less tools therefore less risky for HIV transmission.Latar belakang: Isolat Candida mudah diambil dengan cara mengusap lesi

  20. On Cultural Words, Cultural Meaning Words and Its Translation Method%浅议文化词和文化含义词及其翻译方法

    Institute of Scientific and Technical Information of China (English)

    田苗苗

    2014-01-01

    语言和文化的关系是辩证统一的,翻译过程中蕴含在语言中的文化因素给译者的工作带来了挑战。文章介绍了文化词和文化含义词的各自定义及区别,并对二者在英汉互译中的处理提出了几种方法。%The relationship between language and culture is a dialectical unity, and the cultural factors contained in translation process bring challenges to translator's work. This paper introduces the definitions and differences between cultural words and cultural meaning words, proposes several method in translation.

  1. Detection of streptomycetes in house dust--comparison of culture and PCR methods.

    Science.gov (United States)

    Rintala, H; Hyvärinen, A; Paulin, L; Nevalainen, A

    2004-04-01

    Streptomycetes are gram-positive, spore producing, filamentous bacteria common in soil, but also present in indoor environments. They are potent producers of secondary metabolites and inducers of inflammatory responses in vitro. Polymerase chain reaction (PCR)- and culture-based detection methods for streptomycetes in house dust samples were compared. A total of 47 dust samples were investigated, and the presence of streptomycetes was determined by cultivation on tryptone-yeast-extract-glucose agar and PCR. The 16S rRNA gene of actinomycete isolates from house dust was partially sequenced to investigate if they belong to the genus Streptomyces. Both PCR and culture showed more frequent occurrence of streptomycetes in moisture-damaged homes, although the results did not correlate well. The occurrence of streptomycetes in house dust was associated with moisture damage of the home. The amount of Streptomyces-specific PCR amplification product was significantly higher in dust from moisture-damaged homes than in homes with no moisture damage (P culture, Fisher's exact test). Altogether, the presence of streptomycetes in house dust seems to indicate the presence of moisture damage in the building.

  2. The double pedigree: a method for studying culturally and genetically inherited behavior in tandem.

    Directory of Open Access Journals (Sweden)

    Etienne Danchin

    Full Text Available Transgenerational sources of biological variation have been at the center of evolutionary studies ever since Darwin and Wallace identified natural selection. This is because evolution can only operate on traits whose variation is transmitted, i.e. traits that are heritable. The discovery of genetic inheritance has led to a semantic shift, resulting in the tendency to consider that only genes are inherited across generations. Today, however, concepts of heredity are being broadened again to integrate the accruing evidence of non-genetic inheritance, and many evolutionary biologists are calling for the inclusion of non-genetic inheritance into an inclusive evolutionary synthesis. Here, we focus on social heredity and its role in the inheritance of behavioral traits. We discuss quantitative genetics methods that might allow us to disentangle genetic and non-genetic transmission in natural populations with known pedigrees. We then propose an experimental design based on cross-fostering among animal cultures, environments and families that has the potential to partition inherited phenotypic variation into socially (i.e. culturally and genetically inherited components. This approach builds towards a new conceptual framework based on the use of an extended version of the animal model of quantitative genetics to integrate genetic and cultural components of behavioral inheritance.

  3. The double pedigree: a method for studying culturally and genetically inherited behavior in tandem.

    Science.gov (United States)

    Danchin, Etienne; Pujol, Benoit; Wagner, Richard H

    2013-01-01

    Transgenerational sources of biological variation have been at the center of evolutionary studies ever since Darwin and Wallace identified natural selection. This is because evolution can only operate on traits whose variation is transmitted, i.e. traits that are heritable. The discovery of genetic inheritance has led to a semantic shift, resulting in the tendency to consider that only genes are inherited across generations. Today, however, concepts of heredity are being broadened again to integrate the accruing evidence of non-genetic inheritance, and many evolutionary biologists are calling for the inclusion of non-genetic inheritance into an inclusive evolutionary synthesis. Here, we focus on social heredity and its role in the inheritance of behavioral traits. We discuss quantitative genetics methods that might allow us to disentangle genetic and non-genetic transmission in natural populations with known pedigrees. We then propose an experimental design based on cross-fostering among animal cultures, environments and families that has the potential to partition inherited phenotypic variation into socially (i.e. culturally) and genetically inherited components. This approach builds towards a new conceptual framework based on the use of an extended version of the animal model of quantitative genetics to integrate genetic and cultural components of behavioral inheritance.

  4. Comparison of immunochromatography, PCR and culture methods for the detection of Campylobacter bacteria.

    Science.gov (United States)

    Dey, Shuvra Kanti; Nishimura, Shuichi; Okitsu, Shoko; Hayakawa, Satoshi; Mizuguchi, Masashi; Ushijima, Hiroshi

    2012-12-01

    A total of 463 fecal specimens from human patients were tested for the presence of Camphylobacter by a commercial immunochromatography kit (ImmunoCard STAT! CAMPY), polymerase chain reaction (PCR) and conventional bacteriological (CB) methods. Using culture as the standard of reference, the ImmunoCard STAT! CAMPY assay had a sensitivity of 86% and a specificity of 100%. On the other hand, using PCR as the standard of reference, ImmunoCard STAT! CAMPY assay had a sensitivity of 90.5% and a specificity of 100%.

  5. Establishment of alternative culture method for spermatogonial stem cells using knockout serum replacement.

    Directory of Open Access Journals (Sweden)

    Keisuke Aoshima

    Full Text Available Since spermatogonial stem cells (SSCs are capable of both self-renewal and differentiation to daughter cells for subsequent spermatogenesis, the development of an efficient in vitro culture system is essential for studies related to spermatogenesis. Although the currently available system is serum-free and contains only chemically-defined components, it highly relies upon bovine serum albumin (BSA, a component with batch-to-batch quality variations similar to those of fetal bovine serum. Thus, we searched for an alternative BSA-free culture system that preserved the properties of SSCs. In this study, we utilized Knockout Serum Replacement (KSR in the SSC culture medium, as a substitute for BSA. The results demonstrated that KSR supported the continuous growth of SSCs in vitro and the SSC activity in vivo without BSA, in a feeder-cell combination with mouse embryonic fibroblasts. The addition of BSA to KSR further facilitated cell cycle progression, whereas a transplantation assay revealed that the addition of BSA did not affect the number of SSCs in vivo. The combination of KSR with BSA also allowed the elimination of GFRA1 and FGF2, and the reduction of the GDNF concentration from 20 ng/ml to 5 ng/ml, while maintaining the growth rate and the expression of SSC markers. Furthermore, KSR was also useful with SSCs from non-DBA/2 strains, such as C57BL/6 and ICR. These results suggested that KSR is an effective substitute for BSA for long-term in vitro cultures of SSCs. Therefore, this method is practical for various studies related to SSCs, including spermatogenesis and germ stem cell biology.

  6. Cultural border crossing in three urban classrooms: A mixed methods study

    Science.gov (United States)

    Roopnarine, Rupnarain

    This study examined the effects of the instruction of four youth cultural border crossing behaviors: flexibility, being at ease, playfulness, and citizenship as an intervention aimed at helping students to transition across three borders, student to student, student to science, and student to teacher. The research involved 12 ninth- and 10th-grade students in a large urban school district in three diverse classrooms, A, B, and C. Four students in each classroom volunteered for the study. The students in Groups A and B were in 9th grade Living Environment and students in Group B were in 10th grade chemistry. These students participated in this instructional intervention for three months. The study was conducted using both quantitative and qualitative methods based on participant observations, interviews, and questionnaire. The result indicated that there was no significant effect of the cultural border crossing instructions on the students' interactions across the three borders examined. However, the instructions helped Group A and Group B to be more flexible but not group C. Also, the instructions helped Group A to be more playful and at ease but not Group B and C. The instructions also helped Group A to show more citizenship but not Group B and C. In addition, there was no difference between the pretest and posttest cultural bother crossing behavior. Moreover, qualitative data analysis showed that the participants were more flexible, at ease, and playful among peers than across student to teacher and student to science borders. Also, the use of citizenship in the three groups showed no effect on the participants' interaction with peers. Although, the findings showed no effect of cultural border crossing instructions on students' interactions, it is suggested that we continue to find ways to help students feel more comfortable in science.

  7. Using the House-Tree-Person (H-T-P) as a Method to Understanding Individual Cultural Differences among Children.

    Science.gov (United States)

    Lambeth, Pauline J.

    Finding innovative and effective methods to infer the ability of culturally diverse students continues to present a problem for educators. This paper proposes the House-Tree-Person (HTP) projective technique as a way for school educators to gather important data about student functioning without the use of culturally biased instruments. This…

  8. Using the House-Tree-Person (H-T-P) as a Method to Understanding Individual Cultural Differences among Children.

    Science.gov (United States)

    Lambeth, Pauline J.

    Finding innovative and effective methods to infer the ability of culturally diverse students continues to present a problem for educators. This paper proposes the House-Tree-Person (HTP) projective technique as a way for school educators to gather important data about student functioning without the use of culturally biased instruments. This…

  9. An improved culturing method for opiine fruit fly parasitoids and its application to parasitoid monitoring in the field.

    Science.gov (United States)

    Masry, Ayad; Furlong, Michael J; Clarke, Anthony R; Cunningham, John Paul

    2016-09-21

    Good culturing methods play an important role in the study of insect behavior and its application to pest management. Here, we describe and validate a new method for rearing the parasitoid wasp, Diachasmimorpha kraussii, which attacks some of the world's worst fruit fly pests and is an internationally used biological control agent. Our method differs from standard culturing approaches by presenting adult wasps with host-infested artificial media within a "culturing bag," which mimics a natural (fruit) oviposition substrate. In laboratory trials using wild collected D. kraussii, the culturing bag method was compared to the use of host-infested nectarines, and a commonly used laboratory method of presenting host-infested artificial media within Petri dishes. The culturing bag method proved to be a significant improvement on both methods, combining the advantages of high host survival in artificial media with parasitism levels that were the equivalent to those recorded using host-infested fruits. In our field study, culturing bags infested with the Queensland fruit fly, Bactrocera tryoni, and hung in a mixed peach and nectarine orchard proved to be effective "artificial fruits" attracting wild D. kraussii for oviposition. Significantly more adult wasps were reared from the culturing bags compared to field collected fruits. This was shown to be due to higher fruit fly larval density in the bags, as similar percentage parasitism rates were found between the culturing bags and ripe fruits. We discuss how this cheap, time-efficient method could be applied to collecting and monitoring wild D. kraussii populations in orchards, and assist in maintaining genetic variability in parasitoid laboratory cultures.

  10. Temperature-independent pectin gel method for aerobic plate count in dairy and nondairy food products: collaborative study.

    Science.gov (United States)

    Roth, J N

    1988-01-01

    Ten laboratories participated in a collaborative study to compare the pectin-based plate count (PC) Redigel method with the aerobic plate count and standard plate count agar-based standard methods for the estimation of total bacterial counts in 9 different nondairy food and dairy food products. The foods were cream, homogenized milk, raw milk, cheese, raw chicken, raw oysters, frozen broccoli, flour, and spices. Each laboratory analyzed 6 samples (3 sample pairs) of each food group. Counts obtained by the pectin-based plate count and agar-based plate count methods differed significantly (P less than 0.05) only for homogenized milk, where the pectin gel method resulted in higher counts. The actual counts were higher in the pectin gel method in 8 of the 9 food groups. The log means for pectin gel and agar-based media, respectively, for the 9 food groups were: cream 8.106 and 7.844; homogenized milk 8.642 and 8.231; raw milk 8.711 and 8.423; chicken 7.654 and 7.645; oysters 7.201 and 7.180; broccoli 7.102 and 6.798; cheese 8.045 and 8.055; flour 4.112 and 3.988; spice 5.379 and 5.314. The repeatability standard deviations favored the pectin gel method in 6 of the 9 foods tested. The reproducibility standard deviations favored the pectin gel method in 7 of the 9 foods tested. These results strongly support the suitability of the pectin gel method as an alternative to agar-based plate count and other methods for total bacterial counts in nondairy and dairy food products. The pectin gel method has been adopted official first action.

  11. Independent Directors

    DEFF Research Database (Denmark)

    Ringe, Wolf-Georg

    2013-01-01

    This paper re-evaluates the corporate governance concept of ‘board independence’ against the disappointing experiences during the 2007-08 financial crisis. Independent or outside directors had long been seen as an essential tool to improve the monitoring role of the board. Yet the crisis revealed...... that they did not prevent firms' excessive risk taking; further, these directors sometimes showed serious deficits in understanding the business they were supposed to control, and remained passive in addressing structural problems. A closer look reveals that under the surface of seemingly unanimous consensus...... about board independence in Western jurisdictions, a surprising disharmony prevails about the justification, extent and purpose of independence requirements. These considerations lead me to question the benefits of the current system. Instead, this paper proposes a new, ‘functional’ concept of board...

  12. A xeno-free culture method that enhances Wharton's jelly mesenchymal stromal cell culture efficiency over traditional animal serum-supplemented cultures.

    Science.gov (United States)

    Julavijitphong, Suphakde; Wichitwiengrat, Suparat; Tirawanchai, Nednapis; Ruangvutilert, Pornpimol; Vantanasiri, Chanchai; Phermthai, Tatsanee

    2014-05-01

    Mesenchymal stromal cell (MSC) transplantation holds great promise for use in medical therapies. Several key features of MSCs, including efficient cell growth, generation of sufficient cell numbers and safety, as determined by teratoma formation, make MSCs an ideal candidate for clinical use. However, MSCs derived under standard culture conditions, co-cultured with animal by-products, are inappropriate for therapy because of the risks of graft rejection and animal virus transmission to humans. Alternative serum sources have been sought for stem cell production. We demonstrate for the first time that human serum from umbilical cord blood (hUCS) is an effective co-culture reagent for MSC production from Wharton's jelly MSCs (WJMSCs). Ten umbilical cords were used to generate parallel cultures of WJMSC lines under medium supplemented with hUCS and embryonic stem cell-qualified fetal bovine serum. The WJMSC lines from each medium were analyzed and compared with regard to cell line derivation, proliferation ability and characteristic stability. The phenotypic characteristics of WJMSC derived under either medium showed no differences. WJMSC lines derived under hUCS medium displayed comparable primary culture cell outgrowth, lineage differentiation capacity and cell recovery after cryopreservation compared with supplementation with embryonic stem cell-qualified fetal bovine serum medium. However, superior cell proliferation rates and retention of in vitro propagation (>22 passages) were observed in WJMSC cultures supplemented with hUCS. Additionally, more robust population doubling times were observed in hUCS-supplemented cultures. We conclude that hUCS is an efficient and effective serum source for animal product-free WJMSC line production and can generate MSC lines that may be appropriate for therapeutic use. Copyright © 2014 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  13. Targeted metabolomics in cultured cells and tissues by mass spectrometry: method development and validation.

    Science.gov (United States)

    Abdel Rahman, Anas M; Pawling, Judy; Ryczko, Michael; Caudy, Amy A; Dennis, James W

    2014-10-03

    Metabolomics is the identification and quantitation of small bio-molecules (metabolites) in biological samples under various environmental and genetic conditions. Mass spectrometry provides the unique opportunity for targeted identification and quantification of known metabolites by selective reaction monitoring (SRM). However, reproducibility of this approach depends on careful consideration of sample preparation, chemical classes, and stability of metabolites to be evaluated. Herein, we introduce and validate a targeted metabolite profiling workflow for cultured cells and tissues by liquid chromatography-triple quadrupole tandem mass spectrometry. The method requires a one-step extraction of water-soluble metabolites and targeted analysis of central metabolites that include glycolysis, amino acids, nucleotides, citric acid cycle, and the hexosamine biosynthetic pathway. The sensitivity, reproducibility and molecular stability of each targeted metabolite were assessed under experimental conditions. Quantitation of metabolites by peak area ratio was linear with a dilution over a 4 fold dynamic range with minimal deviation R(2)=0.98. Inter- and intra-day precision with cells and tissues had an average coefficient of variation <15% for cultured cell lines, and somewhat higher for mouse liver tissues. The method applied in triplicate measurements readily distinguished immortalized cells from malignant cells, as well as mouse littermates based on their hepatic metabolic profiles.

  14. The relationship between glass ceiling and power distance as a cultural variable by a new method

    Directory of Open Access Journals (Sweden)

    Naide Jahangirov

    2015-12-01

    Full Text Available Glass ceiling symbolizes a variety of barriers and obstacles that arise from gender inequality at business life. With this mind, culture influences gender dynamics. The purpose of this research was to examine the relationship between the glass ceiling and the power distance as a cultural variable within organizations. Gender variable is taken as a moderator variable in relationship between the concepts. In addition to conventional correlation analysis, we employed a new method to investigate this relationship in detail. The survey data were obtained from 109 people working at a research center which operated as a part of the non-profit private university in Ankara, Turkey. The relationship between the variables was revealed by a new method which was developed as an addition to the correlation in survey. The analysis revealed that the female staff perceived the glass ceiling and the power distance more intensely than the male staff. In addition, the medium level relation was determined between the power distance and the glass ceiling perception among female staff.

  15. Native biofilm cultured under controllable condition and used in mediated method for BOD measurement.

    Science.gov (United States)

    Liu, Ling; Deng, Liu; Yong, Daming; Dong, Shaojun

    2011-05-15

    In this article, we developed a native biofilm (NBF) bioreactor used for biochemical oxygen demand mediated method (BOD(Med)). There were two innovations differed from previous BOD(Med) assay. Firstly, the immobilization of microorganisms was adopted in BOD(Med). Secondly, the NBF was introduced for BOD measurement. The NBF bioreactor has been characterized by optical microscopy. A culture condition of NBF with 24h, 35°C and pH 7 was optimized. Furthermore, a measuring condition with 35°C, pH 7 and 55 mM ferricyanide in 1h incubation were optimized. Based on the optimized condition, the real wastewater samples from local sewage treatment plant had been measured. Performances of the NBFs proposed at different culture conditions were recorded for 110 d, and the results indicated that long-term storage stability was obtained. With the proposed method, an uncontaminated native microbial source solution can be obtained from a wastewater treatment plant. In this way, we can ensure that the microbial species of all in the NBF are same as that in the target to be measured.

  16. Study of Arcobacter spp. contamination in fresh lettuces detected by different cultural and molecular methods.

    Science.gov (United States)

    González, Ana; Ferrús, Maria Antonia

    2011-01-31

    Arcobacters are considered potential emerging food and waterborne pathogens. However, there is no data on the presence of Arcobacter spp. in fresh vegetables. Therefore the objective of this research was to study the presence of Arcobacter spp. in fresh lettuces. Fifty fresh lettuces purchased from different local shops in Valencia (Spain) were analyzed. The assay was performed simultaneously by cultural and molecular methods. Isolates were identified by real-time, multiplex PCR and restriction fragment length polymorphism analysis of PCR-amplified DNA fragment (PCR-RFLP). Finally, all the isolates were genotyped using the randomly amplified polymorphic DNA (RAPD-PCR) method. Arcobacter sp. was detected in 10 of the 50 samples (20%) by real-time PCR, being A. butzleri the unique detected species by mPCR. The detection levels obtained by conventional PCR (7 samples/50, 14%) were slightly lower. These seven samples were found to be positive also by culture isolation. All 19 obtained isolates were identified as A. butzleri by multiplex PCR and PCR-RFLP. Great genetic heterogeneity among the isolates was observed by RAPD-PCR profiling. To our knowledge, this is the first study in which Arcobacter spp. is detected in fresh vegetables such as lettuces. Although these foods are generally considered safe, given the large quantities consumed and the fact that further cooking is absent, lettuce could be a source of Arcobacters of public health concern.

  17. Probabilistic conditional independence structures

    CERN Document Server

    Studeny, Milan

    2005-01-01

    Probabilistic Conditional Independence Structures provides the mathematical description of probabilistic conditional independence structures; the author uses non-graphical methods of their description, and takes an algebraic approach.The monograph presents the methods of structural imsets and supermodular functions, and deals with independence implication and equivalence of structural imsets.Motivation, mathematical foundations and areas of application are included, and a rough overview of graphical methods is also given.In particular, the author has been careful to use suitable terminology, and presents the work so that it will be understood by both statisticians, and by researchers in artificial intelligence.The necessary elementary mathematical notions are recalled in an appendix.

  18. Scientific methods and cultural heritage an introduction to the application of materials science to archaeometry and conservation science

    CERN Document Server

    Artioli, Gilberto

    2010-01-01

    Artioli provides an introduction to the methods and rationales of the scientific investigation of cultural heritage materials, with an emphasis placed on the analytical strategies, modes of operation and resulting information rather than on technicalities.

  19. Choosing Independence

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Milo Djukanovic, Prime Minister of Montenegro, won a key referendum May 21 when voters in his tiny, mountainous nation endorsed a plan to split from Serbia and become an independent state. This marked a final step in the breakup of the former Yugoslavia formed by six republics.

  20. A comparative study of cultural methods for the detection of Salmonella in feed and feed ingredients

    Directory of Open Access Journals (Sweden)

    Haggblom Per

    2009-02-01

    Full Text Available Abstract Background Animal feed as a source of infection to food producing animals is much debated. In order to increase our present knowledge about possible feed transmission it is important to know that the present isolation methods for Salmonella are reliable also for feed materials. In a comparative study the ability of the standard method used for isolation of Salmonella in feed in the Nordic countries, the NMKL71 method (Nordic Committee on Food Analysis was compared to the Modified Semisolid Rappaport Vassiliadis method (MSRV and the international standard method (EN ISO 6579:2002. Five different feed materials were investigated, namely wheat grain, soybean meal, rape seed meal, palm kernel meal, pellets of pig feed and also scrapings from a feed mill elevator. Four different levels of the Salmonella serotypes S. Typhimurium, S. Cubana and S. Yoruba were added to each feed material, respectively. For all methods pre-enrichment in Buffered Peptone Water (BPW were carried out followed by enrichments in the different selective media and finally plating on selective agar media. Results The results obtained with all three methods showed no differences in detection levels, with an accuracy and sensitivity of 65% and 56%, respectively. However, Müller-Kauffmann tetrathionate-novobiocin broth (MKTTn, performed less well due to many false-negative results on Brilliant Green agar (BGA plates. Compared to other feed materials palm kernel meal showed a higher detection level with all serotypes and methods tested. Conclusion The results of this study showed that the accuracy, sensitivity and specificity of the investigated cultural methods were equivalent. However, the detection levels for different feed and feed ingredients varied considerably.

  1. 独立·反叛·狂欢——当代中国摇滚文化的审美研究%Independence,rebellion and carnival——The aesthetic research of the contemporary Chinese rock culture

    Institute of Scientific and Technical Information of China (English)

    邢龙

    2012-01-01

    Due to the special cultural,political and economic environment,Chinese rock culture shows characteristics different from the western rock,that is,independence,which is between the official(serious or elite music) and market(popular music).The aesthetic nature of Chinese rock culture is built on this independence;Rebellion and carnival also showed some Chinese characteristics different from those in the West.%由于中国特殊的文化、政治、经济环境,当代中国摇滚文化表现出不同于西方的特点,即介于官方(严肃或者精英音乐)与市场(流行音乐)之间的一种独立性。中国摇滚文化的审美性正是建立在这种独立性之上,在反叛和狂欢这两个特性上表现了一些不同于西方的中国特色。

  2. Trans-Cultural Comparative Literature Method:Using Grammar Trans-lation Techniques and Communicative Language Teaching Effectively

    Institute of Scientific and Technical Information of China (English)

    李晓彤

    2014-01-01

    Trans-Cultural Comparative Literature Method originated from the discovery of common themes and viewpoints as comparing literary texts in different languages. This article explains how the method combine GT and CLT techniques in a lesson plan that engage students with activities that compare and contrast themes and cultural aspects found in two literary texts. It ex-plores worldviews using information and knowledge of literary structures that motivate Chinese students.

  3. Design of columnar quantum dots for polarization-independent emission using 8-band k{center_dot}p method

    Energy Technology Data Exchange (ETDEWEB)

    Andrzejewski, J; Sek, G; Misiewicz, J [Institute of Physics, Wroclaw University of Technology, Wybrze z-dot e Wyspianskiego 27, 50-370 Wroclaw (Poland); O' Reilly, E [Department of Physics, University College Cork and Tyndall National Institute, Lee Maltings, Cork (Ireland); Fiore, A, E-mail: janusz.andrzejewski@pwr.wroc.p [CORBA Research Institute, Eindhoven University of Technology, PO Box 513, 5600 MB Eindhoven (Netherlands)

    2010-09-01

    Control of the polarization of the emitted light can be highly beneficial for certain optoelectronic applications such as optical amplifiers. It has been recently demonstrated experimentally that semiconductor quantum dots with large height to base length aspect ratio are able to emit polarization-independent light from the edge of the wafer. However, analysis of the physics responsible for the observed polarization properties of such nano-objects (like columnar quantum dots or quantum rods) is still rather limited. In particular, the role of the material surrounding the columnar QD on the strain and thus on the polarization properties has not been considered previously. We report here, based on original software, the results of eight-band k{center_dot}p calculations of the electronic and polarization properties of columnar In{sub y}Ga{sub 1-y}As quantum dots (CQD) with high aspect ratio (up to 6) embedded in an In{sub x}Ga{sub 1-x}As/GaAs quantum well. We calculate the relative intensities of transverse-magnetic (TM) and transverse-electric (TE) linear polarized light emitted from the edge of the semiconductor wafer as a function of the two main factors affecting the heavy hole - light hole valence band mixing and hence the polarization dependent selection rules for the optical transitions, namely i) the composition contrast y/x between the dot material and the surrounding well, and ii) the dot aspect ratio. Our numerical results show, in contrast to the previously reported expectations, that the former is the main driving parameter for tuning the polarization properties. This is explained analyzing the biaxial strain in the CQD, based on which it is possible to predict on the TM to TE intensity ratio.

  4. Modified methods for culturing myoblasts of rats: Combination of multi-enzymatic digestion and double purification

    Institute of Scientific and Technical Information of China (English)

    Li Zhang; Wei Wang; Ming Fan; Xiaoping Chen; Shuhong Liu; Liang Sun

    2007-01-01

    BACKGROUND: With developments of tissue engineering and genetic engineering, we aim to culture myoblasts, which are characterized by high purity, high quality and high production, for wide application in neural regeneration researches.OBJECTIVE: To modify traditional dissociation method in order to obtain myoblasts, which are characterized by high purity, high quality and high production, and explore the biological properties under in vitro culture.DESIGN: Observational study.SETTING: Basic Institute of Academy of Military Medical Sciences of Chinese PLA.MATERIALS: Four neonatal Wistar rats of 5 days old, both genders and mean body mass of 10 g were selected in this study. The main reagents and devices were detailed as follows: DMEM medium (Gibco Company), fetus bovine serum (FBS, Hycolne Company), collagenase Ⅱ (Sigma Company), trypsin (Sigma Company), dispase Ⅱ (Sigma Company), desmin antibody (Fuzhou Maixin Company), antibody Ⅱ and ABC kit (Wuhan Boster Biotechnology Company), desk centrifuge (KUBATO, Japan), and inverted phase contrast microscope (LEICA DMIRB, Germany).METHODS: The experiment was carried out in the Basic Institute of Academy of Military Medical Sciences of Chinese PLA from June to October 2006. Neonatal rats were sacrificed under sterile condition to obtain skeletal muscles of limbs, which were washed with cold PBS (containing benzylpenicillin and estreptomicina), and muscular tissue was sheared into pieces. Then, those muscular pieces were added with mixed digestive enzyme (containing 2 g/L collagenase Ⅱ + 5 g/L dispase Ⅱ + 0.28 g/L CaCl2) as twice volume as pieces, dealt with mechanical pipetting for 5 minutes and cultured in CO2 incubator for 10 minutes.The operation was done for three times and the muscular pieces were digested for 45 minutes in total.Moreover, cells were suspended again in order to obtain myoblasts from skeletal muscle of neonatal rats. In addition, myoblasts were purified with differential attachment technique

  5. An independent evaluation of a new method for automated interpretation of lung scintigrams using artificial neural networks

    Energy Technology Data Exchange (ETDEWEB)

    Holst, H.; Jaerund, A.; Evander, E.; Taegil, K.; Edenbrandt, L. [Dept. of Clinical Physiology, Lund Univ. (Sweden); Maare, K. [Dept. of Radiology, Linkoeping Univ. (Sweden); Aastroem, K. [Dept. of Mathematics, Lund Inst. of Technology (Sweden); Ohlsson, M. [Dept. of Theoretical Physics, Lund Univ. (Sweden)

    2001-01-01

    The purpose of this study was to evaluate a new automated method for the interpretation of lung perfusion scintigrams using patients from a hospital other than that where the method was developed, and then to compare the performance of the technique against that of experienced physicians. A total of 1,087 scintigrams from patients with suspected pulmonary embolism comprised the training group. The test group consisted of scintigrams from 140 patients collected in a hospital different to that from which the training group had been drawn. An artificial neural network was trained using 18 automatically obtained features from each set of perfusion scintigrams. The image processing techniques included alignment to templates, construction of quotient images based on the perfusion/template images, and finally calculation of features describing segmental perfusion defects in the quotient images. The templates represented lungs of normal size and shape without any pathological changes. The performance of the neural network was compared with that of three experienced physicians who read the same test scintigrams according to the modified PIOPED criteria using, in addition to perfusion images, ventilation images when available and chest radiographs for all patients. Performances were measured as area under the receiver operating characteristic curve. The performance of the neural network evaluated in the test group was 0.88 (95% confidence limits 0.81-0.94). The performance of the three experienced experts was in the range 0.87-0.93 when using the perfusion images, chest radiographs and ventilation images when available. Perfusion scintigrams can be interpreted regarding the diagnosis of pulmonary embolism by the use of an automated method also in a hospital other than that where it was developed. The performance of this method is similar to that of experienced physicians even though the physicians, in addition to perfusion images, also had access to ventilation images for

  6. An independent evaluation of a new method for automated interpretation of lung scintigrams using artificial neural networks.

    Science.gov (United States)

    Holst, H; Måre, K; Järund, A; Aström, K; Evander, E; Tägil, K; Ohlsson, M; Edenbrandt, L

    2001-01-01

    The purpose of this study was to evaluate a new automated method for the interpretation of lung perfusion scintigrams using patients from a hospital other than that where the method was developed, and then to compare the performance of the technique against that of experienced physicians. A total of 1,087 scintigrams from patients with suspected pulmonary embolism comprised the training group. The test group consisted of scintigrams from 140 patients collected in a hospital different to that from which the training group had been drawn. An artificial neural network was trained using 18 automatically obtained features from each set of perfusion scintigrams. The image processing techniques included alignment to templates, construction of quotient images based on the perfusion/template images, and finally calculation of features describing segmental perfusion defects in the quotient images. The templates represented lungs of normal size and shape without any pathological changes. The performance of the neural network was compared with that of three experienced physicians who read the same test scintigrams according to the modified PIOPED criteria using, in addition to perfusion images, ventilation images when available and chest radiographs for all patients. Performances were measured as area under the receiver operating characteristic curve. The performance of the neural network evaluated in the test group was 0.88 (95% confidence limits 0.81-0.94). The performance of the three experienced experts was in the range 0.87-0.93 when using the perfusion images, chest radiographs and ventilation images when available. Perfusion scintigrams can be interpreted regarding the diagnosis of pulmonary embolism by the use of an automated method also in a hospital other than that where it was developed. The performance of this method is similar to that of experienced physicians even though the physicians, in addition to perfusion images, also had access to ventilation images for

  7. An improved culture method for selective isolation of Campylobacter jejuni from wastewater

    Directory of Open Access Journals (Sweden)

    Jinyong Kim

    2016-08-01

    Full Text Available 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 (qRT-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. In particular, rifampicin supplementation a