Synthesis and Biological Characterization of a New Norbormide Derived Bodipy FL-Conjugated Fluorescent Probe for Cell Imaging

Background: Norbormide (NRB) is a selective rat toxicant endowed with vasoconstrictor activity confined to the rat peripheral arteries. In a recent work we used a fluorescent derivative of NRB (NRB-AF12), obtained by coupling the NBD fluorophore to the parent molecule via a linker, in order to gain information about the possible site of action of the unlabeled compound. We found that NRB-AF12 labeled intracellular organelles in both NRB-sensitive and -insensitive cells and we accordingly proposed its use as a scaffold for the development of a new class of fluorescent probes. In this study, we examined the fluorescent properties of a BODIPY FL-conjugated NRB probe (MC009) developed: (A) to verify if NRB distribution could be influenced by the attached fluorophore; (B) to improve the fluorescent performance of NRB-AF12. Methods: MC009 characteristics were investigated by confocal fluorescence microscopy, in freshly isolated rat caudal artery myocytes (FIRCAM) and in LX2 cells, representative of NRB-sensitive and insensitive cells, respectively. Main results: In both FIRCAM and LX2 cells MC009 stained endoplasmic reticulum, mitochondria, Golgi apparatus and lipid droplets, revealing the same intracellular distribution as NRB-AF12, and, at the same time, had both improved photostability and gave a more intense fluorescent signal at lower concentrations than was possible with NRB-AF12, which resulted in a better and finer visualization of intracellular structures. Furthermore, MC009 was effective in cellular labeling in both living and fixed cells. At the concentration used to stain the cells, MC009 did not show any cytotoxic effect and did not affect the regular progression of cell cycle and division. Conclusions: This study demonstrates that the distribution of fluorescently labeled NRB is not affected by the type of fluorophore attached to the parent compound, supporting the idea that the localization of the fluorescent derivatives may reasonably reflect that of the parent compound. In addition, we observed a marked improvement in the fluorescent properties of BODIPY FL-conjugated NRB (MC009) over its NBD-derived counterpart (NRB-AF12), confirming NRB as a scaffold for the development of new, high performance, non-toxic fluorescent probes for the labeling of intracellular structures in both living and fixed cells

The Dunes Contribution to the Ecosystem Service of Coastal Protection on the Northern Coast of RS

Coastal dunes have a crucial role in offering the ecosystem service of natural coastal protection. This paper investigates the importance of dunes in providing the ecosystem service of natural coastal protection on the North Coast of the State of Rio Grande do Sul (RS) in southern Brazil. The Integrated Valuation of Ecosystem Services and Tradeoffs (InVEST®) Coastal Vulnerability Model was used to quantify the relative susceptibility to coastal erosion and flooding within the study area. First, the model was run to assess the susceptibility to coastal hazards considering the presence of dunes. Then, the model was run again without the presence of dunes. By comparing the results of the two model simulations, it was possible to identify where dunes contribute to reducing coastal exposure, which indirectly reflects their role in providing the ecosystem service of natural coastal protection. There are two main dune fields in the study area, one in the northern coastal sector and one in the southern sector, respectively in the municipalities of Torres and Cidreira. The index of exposure values produced by the two model simulations were very similar for locations along the Torres coastline. In contrast, along the coast of Cidreira, results from the model simulation without dunes produced higher exposure values than the simulation with the presence of dunes, suggesting that the dunes have an important role in offering natural coastal protection. Dunes are a natural barrier against coastal erosion and flooding, but they are not the only factor influencing the ecosystem service of coastal protection. In Torres, for instance, the local topography and the wind patterns contribute to reducing local exposure. The greatest threat in this sector is the urban sprawl

Performance and egg quality of Embrapa 051 laying hens subjected to different feeding programs.

Abstract: This study aimed to evaluate the productive performance and egg quality of Embrapa 051 lineage laying hens compared to a commercial line (Lohmann Brown) when subjected to different feeding programs. Trial consisted of a randomized complete block design, with four treatments: T1 - Lohmann Brown Line receiving 100% basal diet; T2 - Embrapa 051 Line receiving 93% basal diet; T3 - Embrapa 051 line receiving 100% basal diet; and T4 - Embrapa 051 lineage receiving 107% basal diet. The treatments had five replicates each. The basal diet was that recommended by the Lohmann Brown line manual. Variables of bird performance (egg weight, egg production rate, feed conversion per dozen eggs, energy conversion and egg mass) were analyzed, as well as internal egg quality (Haugh unit, yolk color, yolk percentage, and albumen percentage) and external egg quality (specific gravity, shell percentage, and shell thickness). All performance variables presented significant differences (P <0.05) in the three evaluated cycles, comparing lineages. Egg produced by the Embrapa 051 line had the highest percentages of yolk and those of the Lohmann Brown line had the highest percentage of albumen. In conclusion, Embrapa 051 lineage has the potential to produce good quality eggs, as long as it receives the same feeding program of the genetically established lineage. Resumo: O objetivo deste estudo foi avaliar o desempenho produtivo e a qualidade dos ovos da poedeira Embrapa 051, em comparação com uma linhagem comercial (Lohmann Brown), sujeita a diferentes programas alimentares. O delineamento experimental utilizado foi o de blocos completamente casualizados, e os tratamentos foram: T1 ? Linhagem Lohmann Brown recebendo 100% da dieta base; T2 ? Linhagem Embrapa 051 recebendo 93% da dieta base; T3 ? Linhagem Embrapa 051 recebendo 100% da dieta base; e T4 - Linhagem Embrapa 051 recebendo 107% da dieta base. Os tratamentos tiveram cinco repetições cada. A dieta base foi a recomendada pelo manual da linhagem Lohmann Brown. Foram analisadas variáveis de desempenho zootécnico (peso dos ovos, taxa de produção de ovos, conversão alimentar por dúzia de ovo, conversão energética e massa de ovo); qualidade interna (unidade Haugh, coloração da gema, porcentagem de gema e porcentagem de albúmen) e qualidade externa de ovos (gravidade específica, porcentagem de casca e espessura de casca). Todas as variáveis de desempenho apresentaram diferenças significativas (P<0,05) nos três ciclos avaliados, sendo que as aves da linhagem Lohmann Brown apresentaram os maiores valores para essas variáveis. Em relação às variáveis de qualidade interna e externa dos ovos, apenas as porcentagens de gema e de albúmen apresentaram diferença significativa (P<0,05) entre os tratamentos nos três ciclos avaliados. Os ovos produzidos pela linhagem Embrapa 051 apresentaram as maiores porcentagens de gema e os da linhagem Lohmann Brown a maior porcentagem de albúmen. Com o presente trabalho pode-se concluir que a linhagem Embrapa 051 tem potencial para produzir ovos de boa qualidade, desde que receba o mesmo programa alimentar da linhagem geneticamente estabelecida

TRY plant trait database - enhanced coverage and open access

Plant traits-the morphological, anatomical, physiological, biochemical and phenological characteristics of plants-determine how plants respond to environmental factors, affect other trophic levels, and influence ecosystem properties and their benefits and detriments to people. Plant trait data thus represent the basis for a vast area of research spanning from evolutionary biology, community and functional ecology, to biodiversity conservation, ecosystem and landscape management, restoration, biogeography and earth system modelling. Since its foundation in 2007, the TRY database of plant traits has grown continuously. It now provides unprecedented data coverage under an open access data policy and is the main plant trait database used by the research community worldwide. Increasingly, the TRY database also supports new frontiers of trait-based plant research, including the identification of data gaps and the subsequent mobilization or measurement of new data. To support this development, in this article we evaluate the extent of the trait data compiled in TRY and analyse emerging patterns of data coverage and representativeness. Best species coverage is achieved for categorical traits-almost complete coverage for 'plant growth form'. However, most traits relevant for ecology and vegetation modelling are characterized by continuous intraspecific variation and trait-environmental relationships. These traits have to be measured on individual plants in their respective environment. Despite unprecedented data coverage, we observe a humbling lack of completeness and representativeness of these continuous traits in many aspects. We, therefore, conclude that reducing data gaps and biases in the TRY database remains a key challenge and requires a coordinated approach to data mobilization and trait measurements. This can only be achieved in collaboration with other initiatives

An NBD derivative of the selective rat toxicant norbormide as a new probe for living cell imaging

Norbormide (NRB) is a unique compound that acts directly on rat vascular myocytes to trigger a contractile process, through an as yet unknown mechanism, which results in the selective contraction of rat peripheral arteries. To gain insight into the mechanisms involved in NRB rat-selective activity, we investigated the subcellular distribution of NRB-AF12, a nitrobenzoxadiazole (NBD)-derivative of NRB, in living NRB-sensitive and NRB-insensitive cells. In both cell types, NRB-AF12 localized to the endoplasmic reticulum (ER), Golgi apparatus, mitochondria, lysosomes, and endosomes; however, in NRB-sensitive cells, the fluorescence also extended to the plasma membrane. NRB-AF12 was rapidly internalized into the cells, could easily be washed out and then reloaded back into the same cells, all with a high degree of reproducibility. Cells exposed for 24 h to NRB-AF12 did not show apparent signs of toxicity, even at concentrations of the dye (10 μM) much higher than those required for fluorescence labeling (500 ηM). The distribution pattern of NRB-AF12 fluorescence was near identical to that of ER-Tracker® (Er-Tr), a fluorescent derivative of glibenclamide, a known KATP channel blocker. Displacement tests did not demonstrate, but at the same time did not rule out the possibility of a common target for ER-Tr, NRB-AF12, NRB, and glibenclamide. On the basis of these results we hypothesize a common target site for NRB-AF12 and ER-Tr, and a similar target profile for NRB and glibenclamide, and propose NRB-AF12 as an alternative fluorescence probe to ER-Tracker. Furthermore, NRB-based fluorescence derivatives could be designed to selectively label single cellular structures

Estimation of lung vital capacity before and after coronary artery bypass grafting surgery: a comparison of incentive spirometer and ventilometry

<p>Abstract</p> <p>Background</p> <p>Measurement of vital capacity (VC) by spirometry is the most widely used technique for lung function evaluation, however, this form of assessment is costly and further investigation of other reliable methods at lower cost is necessary. Objective: To analyze the correlation between direct vital capacity measured with ventilometer and with incentive inspirometer in patients in pre and post cardiac surgery.</p> <p>Methodology</p> <p>Cross-sectional comparative study with patients undergoing cardiac surgery. Respiratory parameters were evaluated through the measurement of VC performed by ventilometer and inspirometer. To analyze data normality the Kolmogorov-Smirnov test was applied, for correlation the Pearson correlation coefficient was used and for comparison of variables in pre and post operative period Student's t test was adopted. We established a level of ignificance of 5%. Data was presented as an average, standard deviation and relative frequency when needed. The significance level was set at 5%.</p> <p>Results</p> <p>We studied 52 patients undergoing cardiac surgery, 20 patients in preoperative with VC-ventilometer: 32.95 ± 11.4 ml/kg and VC-inspirometer: 28.9 ± 11 ml/Kg, r = 0.7 p < 0.001. In the post operatory, 32 patients were evaluated with VC-ventilometer: 28.27 ± 12.48 ml/kg and VC-inspirometer: 26.98 ± 11 ml/Kg, r = 0.95 p < 0.001. Presenting a very high correlation between the evaluation forms studied.</p> <p>Conclusion</p> <p>There was a high correlation between DVC measures with ventilometer and incentive spirometer in pre and post CABG surgery. Despite this, arises the necessity of further studies to evaluate the repercussion of this method in lowering costs at hospitals.</p