961 resultados para Laboratory techniques


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This thesis describes the stratigraphy, sedimentology and diagenesis of the Pendleside Limestone (Asbian age), a sequence of limestones, shales and dolostones in the Clitheroe area of N. W. England. Field study of 19 measured sections indicates that it was deposited in a rhythmically subsiding basin (Craven Basin) because of movements on the Mid-Craven Fault which was active in Dinantian times. The sequence is up to 190m thick and consists mostly of distal turbidite deposits which have been reworked at horizons when sediment accumulation built up to the wave base. The original depositional fabric and mineralogy of the Pendleside Limestone Group has been extensively modified by diagenetic processes including cementation, authi­genesis, dolomitization and silicification. These processes have been studied using a wide variety of laboratory techniques. The carbonate cements of the PendIeside Limestone consist predominantly of ferroan calcite and non-ferroan calcite with microdolomite incIusions. The former is probably a stable replacement of original-high-magnesian calcite. Cementation was accompanied by the formation of authigenic albite and quartz. Much of the upper part of the Pendleside Limestone has been extensively dolomitized and chertified. Several distinct zones of dolomitization are found which increase in thickness and intensity towards the top of the Pendleside Limestone Group. The dolostone horizons correspond to coarser-grained lithologies deposited during periods of shallow water sedimentation. The composition of the dolomites changes from ferroan dolomite in the lower part of the Group to non-ferroan dolomite in the upper part. The low strontium and sodium content of the dolostones in association with the other evidence suggests that the dolomitization was brought about in an open system by the mixing of marine and fresh water in phreatic lens which were established at periodic intervals. The dolomitization was closely associated with chertification although this was initiated by the dissolution of siliceous spicules which provided the necessary source of silica.

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A major problem with breast cancer treatment is the prevalence of antiestrogen resistance, be it de novo or acquired after continued use. Many of the underlying mechanisms of antiestrogen resistance are not clear, although estrogen receptor-mediated actions have been identified as a pathway that is blocked by antiestrogens. Selective estrogen receptor modulators (SERMs), such as tamoxifen, are capable of producing reactive oxygen species (ROS) through metabolic activation, and these ROS, at high levels, can induce irreversible growth arrest that is similar to the growth arrest incurred by SERMs. This suggests that SERM-mediated growth arrest may also be through ROS accumulation. Breast cancer receiving long-term antiestrogen treatment appears to adapt to this increased, persistent level of ROS. This, in turn, leads to the disruption of reversible redox signaling that involves redox-sensitive phosphatases and protein kinases and transcription factors. This has downstream consequences for apoptosis, cell cycle progression, and cell metabolism. For this dissertation, we explored if altering the ROS formed by tamoxifen also alters sensitivity of the drug in resistant cells. We explored an association with a thioredoxin/Jab1/p27 pathway, and a possible role of dysregulation of thioredoxin-mediated redox regulation contributing to the development of antiestrogen resistance in breast cancer. We used standard laboratory techniques to perform proteomic assays that showed cell proliferation, protein concentrations, redox states, and protein-protein interactions. We found that increasing thioredoxin reductase levels, and thus increasing the amount of reduced thioredoxin, increased tamoxifen sensitivity in previously resistant cells, as well as altered estrogen and tamoxifen-induced ROS. We also found that decreasing levels of Jab1 protein also increased tamoxifen sensitivity, and that the downstream effects showed a decrease p27 phosphorylation in both cases. We conclude that the chronic use of tamoxifen can lead to an increase in ROS that alters cell signaling and causing cell growth in the presence of tamoxifen, and that this resistant cell growth can be reversed with an alteration to the thioredoxin/Jab1 pathway.

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The microorganisms play very important roles in maintaining ecosystems, which explains the enormous interest in understanding the relationship between these organisms as well as between them and the environment. It is estimated that the total number of prokaryotic cells on Earth is between 4 and 6 x 1030, constituting an enormous biological and genetic pool to be explored. Although currently only 1% of all this wealth can be cultivated by standard laboratory techniques, metagenomic tools allow access to the genomic potential of environmental samples in a independent culture manner, and in combination with third generation sequencing technologies, the samples coverage become even greater. Soils, in particular, are the major reservoirs of this diversity, and many important environments around us, as the Brazilian biomes Caatinga and Atlantic Forest, are poorly studied. Thus, the genetic material from environmental soil samples of Caatinga and Atlantic Forest biomes were extracted by direct techniques, pyrosequenced, and the sequences generated were analyzed by bioinformatics programs (MEGAN MG-RAST and WEBCarma). Taxonomic comparative profiles of the samples showed that the phyla Proteobacteria, Actinobacteria, Acidobacteria and Planctomycetes were the most representative. In addition, fungi of the phylum Ascomycota were identified predominantly in the soil sample from the Atlantic Forest. Metabolic profiles showed that despite the existence of environmental differences, sequences from both samples were similarly placed in the various functional subsystems, indicating no specific habitat functions. This work, a pioneer in taxonomic and metabolic comparative analysis of soil samples from Brazilian biomes, contributes to the knowledge of these complex environmental systems, so far little explored

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Vesiculoviruses (VSV) are zoonotic viruses that cause vesicular stomatitis disease in cattle, horses and pigs, as well as sporadic human cases of acute febrile illness. Therefore, diagnosis of VSV infections by reliable laboratory techniques is important to allow a proper case management and implementation of strategies for the containment of virus spread. We show here a sensitive and reproducible real-time reverse transcriptase polymerase chain reaction (RT-PCR) for detection and quantification of VSV. The assay was evaluated with arthropods and serum samples obtained from horses, cattle and patients with acute febrile disease. The real-time RT-PCR amplified the Piry, Carajas, Alagoas and Indiana Vesiculovirus at a melting temperature 81.02 ± 0.8ºC, and the sensitivity of assay was estimated in 10 RNA copies/mL to the Piry Vesiculovirus. The viral genome has been detected in samples of horses and cattle, but not detected in human sera or arthropods. Thus, this assay allows a preliminary differential diagnosis of VSV infections.

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Résumé : Cette recherche descriptive a pour origine une question de praticienne-chercheure : Quels sont les stratégies et les actes pédagogiques à privilégier par un enseignant en sciences au collégial, afin de stimuler la motivation des étudiants à s’engager dans leurs activités d’apprentissage? Les enjeux pédagogiques au niveau collégial sont d’importance. Les études scientifiques sont caractérisées par un grand nombre d’abandons, un faible niveau de motivation et d’engagement des étudiants dans leurs études et un taux de réussite sous les seuils visés. L’enseignant peut jouer un rôle-clef pour stimuler l’engagement des étudiants, tant par ses stratégies pédagogiques ou ses choix d’activité d’apprentissage que par le support qu’il peut apporter à l’apprenant. La particularité du milieu collégial, où se côtoient formation pré-universitaire et formation technique, n’a pas encore été étudiée, dans une perspective comparative sur la dynamique motivationnelle, en lien avec l’engagement dans une formation scientifique. L’objectif général de cette recherche est de décrire les perceptions des étudiants quant à leur motivation, selon différents actes et stratégies pédagogiques mis en place par les enseignants de deux programmes scientifiques au niveau collégial, soit le programme pré-universitaire Sciences de la nature et pour le programme Techniques de laboratoire, profil Biotechnologies. Chacun des 16 enseignants participants de biologie ou de physique des deux programmes ont été observés durant une ou deux séances de cours totalisant 58 heures d’enseignement, pour 29 groupes d’étudiants, de façon à identifier la stratégie pédagogique générale de la séance et les actes pédagogiques qui ont été concrétisés en classe. Les étudiants ont ensuite complété un questionnaire en ligne sur leurs perceptions en lien avec la motivation concernant ces stratégies et actes pédagogiques. Les questionnaires de 272 répondants ont été traité, analysés et interprétés, de façon à dresser un portrait de la dynamique motivationnelle par groupes de comparaison, par analyses statistiques et analyse qualitative des commentaires spontanés des répondants. Les étudiantes et étudiants de Biotechnologies ont des perceptions plus favorables à la motivation en accordant plus de valeur aux activités d’apprentissage que les répondants de Sciences de la nature. Ils ont également un désir d’engagement plus élevé. Les hommes de Biotechnologies se sentent significativement plus compétents que les femmes du même programme et les répondants des deux genres en Sciences de la nature, ils ont une plus grande facilité à participer aux activités d’apprentissage en classe et ne mentionnent que très peu d’émotions négatives en lien avec les activités de leurs séances de cours. Les étudiantes et étudiants des deux programmes ont des perceptions très diversifiées sur les activités d’apprentissage et stratégies pédagogiques; elles sont même opposées entre certains étudiants pour une même séance de cours, ce qui indique que d’offrir une bonne variété d’actes pédagogiques à chaque séance de cours est à privilégier pour stimuler la motivation du plus grand nombre.

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Laboratory colonies of 15 economically important species of multi-host fruit flies (Diptera:Tephritidae) have been established in eight South Pacific island countries for the purpose of undertaking biological studies, particularly host status testing and research on quarantine treatments. Laboratory rearing techniques are based on the development of artificial diets for larvae consisting predominately of the pulp of locally available fruits including pawpaw, breadfruit and banana. The pawpaw diet is the standard diet and is used in seven countries for rearing 11 species. Diet ingredients are standard proportions of fruit pulp, hydrolysed protein and a bacterial and fungal inhibitor. The diet is particularly suitable for post-harvest treatment studies when larvae of known age are required. Another major development in the laboratory rearing system is the use of pure strains of Enterobacteriaceae bacterial cultures as important adult-feeding supplements. These bacterial cultures are dissected out of the crop of wild females, isolated by sub-culturing, and identified before supply to adults on peptone yeast extract agar plates. Most species are egged using thin, plastic receptacles perforated with 1 mm oviposition holes, with fruit juice or larval diet smeared internally as an oviposition stimulant. Laboratory rearing techniques have been standardised for all of the Pacific countries. Quality control monitoring is based on acceptable ranges in per cent egg hatch, pupal weight and pupal mortality. Colonies are rejuvenated every 6 to 12 months by crossing wild males with laboratory-reared females and vice versa. The standard rearing techniques, equipment and ingredients used in collecting, establishment, maintenance and quality control of these fruit fly species are detailed in this paper.

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A detailed study on the removal of pollutants (NOx, aldehydes and CO) from the exhaust of a stationary diesel engine is carried out using barrier discharge hybrid plasma techniques. The objective of the study is to make a comparative analysis. For this purpose, the exhaust treatment was carried out in two stages. In the first stage, the exhaust was treated with plasma process and plasma-adsorbent hybrid process. The effectiveness of the two processes with regard to NOx removal and by-product reduction was discussed. In the second stage, the exhaust was treated by plasma and plasma-catalyst hybrid process. The effectiveness of the two processes with regard to pollutants (NOx, CO) removal and by-product reduction was analyzed. Finally, a comprehensive comparison of different techniques has been made and feasible plasma based hybrid techniques for stationary and non-stationary engine exhaust treatments were proposed.

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The current paper presents a study conducted at CERN, Switzerland, to investigate visitors' and tour guides' use and appreciation of existing panels at visit itinerary points. The results were used to develop a set of recommendations for constructing optimal panels to assist the guides' explanation.

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In healthy individuals, Candida species are considered commensal yeasts of the oral cavity. However, these microorganisms can also act as opportunist pathogens, particularly the so-called non-albicans Candida species that are increasingly recognized as important agents of human infection. Several surveys have documented increased rates of C. glabrata, C. tropicalis, C. guilliermondii, C. dubliniensis, C. parapsilosis, and C. krusei in local and systemic fungal infections. Some of these species are resistant to antifungal agents. Consequently, rapid and correct identification of species can play an important role in the management of candidiasis. Conventional methods for identification of Candida species are based on morphological and physiological attributes. However, accurate identification of all isolates from clinical samples is often complex and time-consuming. Hence, several manual and automated rapid commercial systems for identifying these organisms have been developed, some of which may have significant sensitivity issues. To overcome these limitations, newer molecular typing techniques have been developed that allow accurate and rapid identification of Candida species. This study reviewed the current state of identification methods for yeasts, particularly Candida species. © 2013 John Wiley & Sons A/S.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Protocol for the Genetics Laboratory practices.

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Slides for the Genetics Laboratory practices introduction.