Trypan blue exclusion assay by flow cytometry

Dye exclusion tests are used to determine the number of live and dead cells. These assays are based on the principle that intact plasma membranes in live cells exclude specific dyes, whereas dead cells do not. Although widely used, the trypan blue (TB) exclusion assay has limitations. The dye can be incorporated by live cells after a short exposure time, and personal reliability, related to the expertise of the analyst, can affect the results. We propose an alternative assay for evaluating cell viability that combines the TB exclusion test and the high sensitivity of the flow cytometry technique. Previous studies have demonstrated the ability of TB to emit fluorescence when complexed with proteins. According to our results, TB/bovine serum albumin and TB/cytoplasmic protein complexes emit fluorescence at 660 nm, which is detectable by flow cytometry using a 650-nm low-pass band filter. TB at 0.002% (w/v) was defined as the optimum concentration for distinguishing unstained living cells from fluorescent dead cells, and fluorescence emission was stable for 30 min after cell treatment. Although previous studies have shown that TB promotes green fluorescence quenching, TB at 0.002% did not interfere with green fluorescence in human live T-cells stained with anti-CD3/fluorescein isothiocyanate (FITC) monoclonal antibody. We observed a high correlation between the percentage of propidium iodide+CD3/FITC+ and TB+CD3/FITC+ cells, as well as similar double-stained cell profiles in flow cytometry dot-plot graphs. Taken together, the results indicate that a TB exclusion assay by flow cytometry can be employed as an alternative tool for quick and reliable cell viability analysis.

Kaukolämpöjohdon lämpöhäviöiden talteenotto lämpöpumpputeknologian avulla

Tämän tutkimuksen tavoitteena oli laatia tarkastelumalli, jonka perusteella pystyttäisiin analysoimaan tutkimuksessa tarkasteltavan kaukolämpöjohdon lämpöhäviöiden talteenot-toratkaisun taloudellista kannattavuutta yleisellä tasolla sekä sen mahdollisissa sovellus-kohteissa. Työssä tarkastellaan kaukolämpöjohtoa, jonka sisään on sijoitettu lämmönke-ruuputki. Lämmönkeruuputken on tarkoitus kerätä lämpöä kaukolämpöjohdon vaipasta sekä pitää vaipan lämpötilaa ympäristön lämpötilaa matalampana, jolloin johdon ulkopuo-lisia lämpöhäviöitä ei synny. Tarkastelumalli laadittiin perustuen lämpöpumppuprosessin ja kaukolämpöverkoston yleisiin mitoitusperiaatteisiin sekä ratkaisuun liittyvien järjestelmien osalta kerättyihin tarkas-teluhetkeä edustaviin kustannustietoihin. Tarkastelumallista laadittiin Excel-laskentataulukkona, jota voidaan tulevaisuudessa soveltaa järjestelmän sovelluskohdekoh-taiseen tarkasteluun sekä mitoitukseen. Lasketut takaisinmaksuajat osoittautuivat kaikissa tarkastelluissa tapauksissa järjestelmien arvioitua teknistä käyttöikää lyhyemmäksi. Järjestelmällä voisi olla tietynlaisissa sovellus-kohteissa myös strateginen, kaukolämpöliiketoiminnan riskejä vähentävä merkitys.

Modélisation de l'influence de la fragmentation des habitats sur le risque de prédation chez le caribou forestier

Les populations du caribou forestier (Rangifer tarandus caribou) sont en déclin sur l’ensemble de leur aire de répartition en Amérique du Nord. Il s’avère que la prédation, amplifiée par l’exploitation forestière, en est la principale cause. Ce projet consiste à mettre en place un outil d'aide à la décision, permettant de modéliser les changements du risque de prédation chez le caribou forestier durant la succession forestière, et ce, selon différents scénarios d'aménagement forestier simulés sur la Côte-Nord du Québec, Canada. Ces scénarios, simulés de 2000 à 2150, sont caractérisés par (i) des coupes limitées aux blocs de protection et de remplacement, (ii) des coupes étendues sur le paysage, et finalement (iii) par l'absence de coupe dès 2000. Un modèle basé sur l'individu (MBI) permet de modéliser les déplacements simultanés du caribou forestier, de l'orignal (Alces alces) et du loup gris (Canis lupus) afin d'évaluer le risque de prédation selon les scénarios. En général, le risque de prédation est plus important lorsque les coupes sont limitées aux blocs de protection et de remplacement. En effet, il semble que ces blocs augmentent les probabilités de rencontre entre les proies et leurs prédateurs. Toutefois, certains résultats ne reflètent pas la littérature, ce qui montre la nécessité d'améliorer le MBI. Certaines recommandations visent finalement à bonifier ce MBI pour permettre l'analyse de la viabilité à long terme du caribou forestier en fonction de différents scénarios d'aménagement forestier.

Evaluating the Small Population Paradigm for Rare Large-Bodied Woodpeckers, with Implications for the Ivory-billed Woodpecker

Six large-bodied, ≥ 120 g, woodpecker species are listed as near-threatened to critically endangered by the International Union for Conservation of Nature (IUCN). The small population paradigm assumes that these populations are likely to become extinct without an increase in numbers, but the combined influences of initial population size and demographic rates, i.e., annual adult survival and fecundity, may drive population persistence for these species. We applied a stochastic, stage-based single-population model to available demographic rates for Dryocopus and Campephilus woodpeckers. In particular, we determined the change in predicted extinction rate, i.e., proportion of simulated populations that went extinct within 100 yr, to concomitant changes in six input parameters. To our knowledge, this is the first study to evaluate the combined importance of initial population size and demographic rates for the persistence of large-bodied woodpeckers. Under a worse-case scenario, the median time to extinction was 7 yr (range: 1–32). Across the combinations of other input values, increasing initial population size by one female induced, on average, 0.4%–3.2% (range: 0%–28%) reduction in extinction rate. Increasing initial population size from 5–30 resulted in extinction rates < 0.05 under limited conditions: (1) all input values were intermediate, or (2) Allee effect present and annual adult survival ≥ 0.8. Based on our model, these species can persist as rare, as few as five females, and thus difficult-to-detect, populations provided they maintain ≥ 1.1 recruited females annually per adult female and an annual adult survival rate ≥ 0.8. Athough a demographic-based population viability analysis (PVA) is useful to predict how extinction rate changes across scenarios for life-history attributes, the next step for modeling these populations should incorporate more easily acquired data on changes in patch occupancy to make predictions about patch colonization and extinction rates.

Stability of a Seabird Population in the Presence of an Introduced Predator

We hypothesized that although large populations may appear able to withstand predation and disturbance, added stochasticity in population growth rate (λ) increases the risk of dramatic population declines. Approximately half of the Aleutian Islands' population of Least Auklets (Aethia pusilla) breed at one large colony at Kiska Island in the presence of introduced Norway rats (Rattus norvegicus) whose population erupts periodically. We evaluated two management plans, do nothing or eradicate rats, for this colony, and performed stochastic elasticity analysis to focus future research and management. Our results indicated that Least Auklets breeding at Kiska Island had the lowest absolute value of growth rate and more variable λ's (neither statistically significant) during 2001-2010, when compared with rat-free colonies at Buldir and Kasatochi islands. We found variability in the annual proportional change in population size among islands with Kiska Island having the fastest rate of decline, 78% over 20 years. Under the assumption that the eradication of rats would result in vital rates similar to those observed at rat-free Buldir and Kasatochi islands, we found the projected population decline decreased from 78% to 24% over 20 years. Overall, eradicating rats at Kiska Island is not likely to increase Least Auklet vital rates, but will decrease the amount of variation in λ, resulting in a significantly slower rate of population decline. We recommend the eradication of rats from Kiska Island to decrease the probability of dramatic population declines and ensure the future persistence of this important colony.

Endodontic Chelators Induce Nitric Oxide Expression by Murine-cultured Macrophages

Introduction: Endodontic chelators may extrude to apical tissues during instrumentation activating cellular events on periapical tissues. This study assessed in vitro the expression of nitric oxide (NO) concentrations by murine peritoneal macrophages after contact with MTAD (Dentsply/Tulsa, Tulsa, OK), Tetraclean (Ogna Laboratori Farmaceutici, Muggio, Italy), Smear Clear (Sybron Endo, Orange, CA), and EDTA (Biodinamica, Ibipora, PR, Brazil). Methods: Macrophage cells were obtained from Swiss mice after peritoneal lavage. Chelators were diluted in distilled water obtaining 12 concentrations, and MTT assay identified the concentrations, per group, displaying the highest cell viability (analysis of variance, p < 0.01). Selected concentrations were tested for NO expression using Griess reaction. Culture medium and lipopolysaccharide (LPS) were used as controls. Results: Analysis of variance and Tukey tests showed that all chelators displayed elevated NO concentrations compared with the negative control (p < 0.01). MTAD induced the lowest NO expression, followed by Tetraclean, EDTA, and Smear Clear. No difference was observed between MTAD and Tetraclean (p > 0.01), Tetraclean and EDTA (p > 0.01), and EDTA and Smear Clear (p > 0.01). LPS ranked similar to both EDTA and Smear Clear (p > 0.01). Conclusion: The tested endodontic chelators displayed severe proinflammatory effects on murine-cultured macrophages. Citric acid-based solutions induce lower No release than EDTA-based irrigants. (J Endod 2009;35:824-828)

Effects of species turnover on reserve site selection in a fragmented landscape

Changes in species composition is an important process in many ecosystems but rarely considered in systematic reserve site selection. To test the influence of temporal variability in species composition on the establishment of a reserve network, we compared network configurations based on species data of small mammals and frogs sampled during two consecutive years in a fragmented Atlantic Forest landscape (SE Brazil). Site selection with simulated annealing was carried out with the datasets of each single year and after merging the datasets of both years. Site selection resulted in remarkably divergent network configurations. Differences are reflected in both the identity of the selected fragments and in the amount of flexibility and irreplaceability in network configuration. Networks selected when data for both years were merged did not include all sites that were irreplaceable in one of the 2 years. Results of species number estimation revealed that significant changes in the composition of the species community occurred. Hence, temporal variability of community composition should be routinely tested and considered in systematic reserve site selection in dynamic systems.

Damages of spittlebug on sugarcane quality and fermentation process

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

Viabilidade econômica do cultivo de shiitake em diferentes escalas de produção

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

Custos e viabilidade econômica da produção de lambari-do-rabo amarelo em Monte Castelo/SP: um estudo de caso

The aim of this study was to analyze the cost of production and economic viability of production of Lambari in Monte Castelo/SP, with the primary market for the live-bait for sport fishing of carnivorous fish. To calculate the operating cost structure was used the model proposed by the Instituto de Economia Agricola from São Paulo/Brazil. In analyzing the economic viability of investment, we determined the Internal Rate of Return (IRR) and Pay Back Period. It was noted that the inputs represented approximately 74.0% of the EOC (Effective Operating Cost). In the rentability analysis (short term), there was an operating profit of R$ 3,133.45 for a cycle, with a profitability of 18.65% for four months of production, a cost price of R$ 13.02 kg(-1). For viability of the activity (long term), the results were attractive to recovery capital from the 3(rd) year (Pay Back), with an IRR of 25.68% and 89.0% return on the producer in 5 years (BCR - benefit cost ratio). The study of costs and economic viability analysis reinforce the importance of rational management in the productive system and the importance of planning and managing the profitability of the activity.

Endodontic Chelators Induce Nitric Oxide Expression by Murine-cultured Macrophages

Introduction: Endodontic chelators may extrude to apical tissues during instrumentation activating cellular events on periapical tissues. This study assessed in vitro the expression of nitric oxide (NO) concentrations by murine peritoneal macrophages after contact with MTAD (Dentsply/Tulsa, Tulsa, OK), Tetraclean (Ogna Laboratori Farmaceutici, Muggio, Italy), Smear Clear (Sybron Endo, Orange, CA), and EDTA (Biodinamica, Ibipora, PR, Brazil). Methods: Macrophage cells were obtained from Swiss mice after peritoneal lavage. Chelators were diluted in distilled water obtaining 12 concentrations, and MTT assay identified the concentrations, per group, displaying the highest cell viability (analysis of variance, p < 0.01). Selected concentrations were tested for NO expression using Griess reaction. Culture medium and lipopolysaccharide (LPS) were used as controls. Results: Analysis of variance and Tukey tests showed that all chelators displayed elevated NO concentrations compared with the negative control (p < 0.01). MTAD induced the lowest NO expression, followed by Tetraclean, EDTA, and Smear Clear. No difference was observed between MTAD and Tetraclean (p > 0.01), Tetraclean and EDTA (p > 0.01), and EDTA and Smear Clear (p > 0.01). LPS ranked similar to both EDTA and Smear Clear (p > 0.01). Conclusion: The tested endodontic chelators displayed severe proinflammatory effects on murine-cultured macrophages. Citric acid-based solutions induce lower No release than EDTA-based irrigants. (J Endod 2009;35:824-828)

Modeling of a MIT for the application of a frequency inverter of the electric vehicle

Nowadays, drives that use a combination of induction motors and frequency inverters are very common, a fact due to the financial practicality and viability in purchasing and operating that system. This system modeling and simulation becomes important when it wants to evaluate the performance, to calculate and correct parameters, and it has a fundamental role in functionality and viability analysis for application of new configurations and technologies. This work is about to elaborate a simple induction motor model based in the torque versus speed characteristic, using the linearization method for application in a specific operation range to be controlled by a frequency inverter. © 2011 Springer-Verlag Berlin Heidelberg.

Análises técnica e energética da secagem combinada no processamento de casulo do bicho-da-seda de Bombyx Mori L

Pós-graduação em Agronomia (Energia na Agricultura) - FCA

O uso do caleidoscópio no ensino de grupos de simetria e transformações geométricas

Pós-graduação em Educação Matemática - IGCE

Análise de viabilidade econômica de um aterro sanitário para cidade de pequeno porte

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)