A comprehensive high-throughput FTIR spectroscopy-based method for evaluating the transfection event: estimating the transfection efficiency and extracting associated metabolic responses

Reporter genes are routinely used in every laboratory for molecular and cellular biology for studying heterologous gene expression and general cellular biological mechanisms, such as transfection processes. Although well characterized and broadly implemented, reporter genes present serious limitations, either by involving time-consuming procedures or by presenting possible side effects on the expression of the heterologous gene or even in the general cellular metabolism. Fourier transform mid-infrared (FT-MIR) spectroscopy was evaluated to simultaneously analyze in a rapid (minutes) and high-throughput mode (using 96-wells microplates), the transfection efficiency, and the effect of the transfection process on the host cell biochemical composition and metabolism. Semi-adherent HEK and adherent AGS cell lines, transfected with the plasmid pVAX-GFP using Lipofectamine, were used as model systems. Good partial least squares (PLS) models were built to estimate the transfection efficiency, either considering each cell line independently (R 2 ≥ 0.92; RMSECV ≤ 2 %) or simultaneously considering both cell lines (R 2 = 0.90; RMSECV = 2 %). Additionally, the effect of the transfection process on the HEK cell biochemical and metabolic features could be evaluated directly from the FT-IR spectra. Due to the high sensitivity of the technique, it was also possible to discriminate the effect of the transfection process from the transfection reagent on KEK cells, e.g., by the analysis of spectral biomarkers and biochemical and metabolic features. The present results are far beyond what any reporter gene assay or other specific probe can offer for these purposes.

Compression Efficiency Analysis of Wyner-Ziv Video Coding with Motion Compensated Side Information Interpolation

The Wyner-Ziv video coding (WZVC) rate distortion performance is highly dependent on the quality of the side information, an estimation of the original frame, created at the decoder. This paper, characterizes the WZVC efficiency when motion compensated frame interpolation (MCFI) techniques are used to generate the side information, a difficult problem in WZVC especially because the decoder only has available some reference decoded frames. The proposed WZVC compression efficiency rate model relates the power spectral of the estimation error to the accuracy of the MCFI motion field. Then, some interesting conclusions may be derived related to the impact of the motion field smoothness and the correlation to the true motion trajectories on the compression performance.

Fluorescence Microscopy Imaging Denoising with Log-Euclidean Priors and Photobleaching Compensation

Fluorescent protein microscopy imaging is nowadays one of the most important tools in biomedical research. However, the resulting images present a low signal to noise ratio and a time intensity decay due to the photobleaching effect. This phenomenon is a consequence of the decreasing on the radiation emission efficiency of the tagging protein. This occurs because the fluorophore permanently loses its ability to fluoresce, due to photochemical reactions induced by the incident light. The Poisson multiplicative noise that corrupts these images, in addition with its quality degradation due to photobleaching, make long time biological observation processes very difficult. In this paper a denoising algorithm for Poisson data, where the photobleaching effect is explicitly taken into account, is described. The algorithm is designed in a Bayesian framework where the data fidelity term models the Poisson noise generation process as well as the exponential intensity decay caused by the photobleaching. The prior term is conceived with Gibbs priors and log-Euclidean potential functions, suitable to cope with the positivity constrained nature of the parameters to be estimated. Monte Carlo tests with synthetic data are presented to characterize the performance of the algorithm. One example with real data is included to illustrate its application.

Reward-risk efficiency in proportional reinsurance with different risk measures

We have studied, in particular under normality of the implied random variables, the connections between different measures of risk such as the standard deviation, the W-ruin probability and the p-V@R. We discuss conditions granting the equivalence of these measures with respect to risk preference relations and the equivalence of dominance and efficiency of risk-reward criteria involving these measures. Then more specifically we applied these concepts to rigorously face the problem of finding the efficient set of de Finetti’s variable quota share proportional reinsurance.

From efficiency to optimality in proportional reinsurance under group correlation

Based on our recent discovery of closed form formulae of efficient Mean Variance retentions in variable quota-share proportional reinsurance under group correlation, we analyzed the influence of different combination of correlation and safety loading levels on the efficient frontier, both in a single period stylized problem and in a multiperiod one.

Light pollution and energy efficiency: a case study of the village of Vialonga

Since industrialization and the formation of larger urban centers in the nineteenth century, pollution of the environment was always present in daily life in various ways, namely in the form of light. Light pollution can cause various consequences, both for humans and for their ecosystem, producing effects on environmental, social, economic and scientific level. In Portugal, the lighting is responsible for 3% of total electricity consumption, energy costs are in some cases more than 50% towards the costs incurred by municipalities with energy, checking-in recent years a trend similar to that improvement of illumination levels in the region (about 4 to 5% per year). Proper use of lighting brings many benefits both to the citizen and environment, since greater energy efficiency can contribute to reducing CO2 emissions, energy costs, as well as to decrease the use of resources not-renewable and/or contamination of renewable resources, which can occurs in the process of obtaining electricity. The present study has a main goal to analyze the illuminance levels associated to the public lighting of the village of Vialonga, Vila Franca de Xira (Portugal), to verify if it is efficient. The aim is also to relate the efficiency of street lighting with the existence of light pollution.

Biological monitoring of aflatoxin (AFB1) in workers of swine and poultry production

Although a great body of literature exists concerning the ingestion of food contaminated with aflatoxin, there are still few studies regarding mycotoxin inhalation in occupational settings. Since mycotoxins are relatively non-volatile, inhalation exposure is cause by inhalation of airborne fungal particulates or fungi-contaminated substrates that contain aflatoxin. We intend to know if there is occupational exposure to aflatoxin in Portuguese poultry and swine production. A total of 19 individuals (11 swine; 8 poultry) agreed and provided blood samples during the course of this investigation. Measurement of AFB1 was performed by ELISA. The samples were treated with pronase (Merck), wash in a Column C18 and purification was made with immunoaffinity columns (R.biopharma), specific for AFB1. It was applied statistical test (Mann-Whitney) to verified statistical difference in AFB1 results between the two settings. Results varied with concentrations from

Articulação do tratamento das águas lixiviantes de infraestruturas de resíduos com ETAR urbanas: caso de estudo

A deposição de resíduos sólidos urbanos (RSU) em aterros sanitários (AS) tem uma dimensão que necessita de uma atenção especial por parte da sociedade. Deste acto podem resultar situações gravosas para o meio ambiente se não forem tomadas as medidas correctas. Um dos pontos que necessita de uma particular atenção é o tratamento das águas lixiviantes dos aterros sanitários, uma vez que estas apresentam geralmente cargas altamente poluentes. O presente trabalho apresenta a estrutura de um aterro sanitário, definindo os seus princípios de funcionamento, bem como as principais características a ter em consideração para a sua concepção e construção. É ainda abordada a composição dos resíduos sólidos produzidos em Portugal de acordo com (1) e as características quantitativas e qualitativas do lixiviado produzido por um AS. São apresentados os sistemas tratamento convencionais, biológicos e físico-químicos, mais utilizados em Portugal para o tratamento de águas lixiviantes, relacionando as problemáticas e as limitações associadas a cada um deles. É ainda apresentado o tratamento combinado de águas lixiviantes com águas residuais e a recirculação de lixiviado no sistema de tratamento, assim como algumas das vantagens associadas a estas práticas. Tendo por base, uma situação real, avalia-se um caso de estudo, que visa a avaliação do funcionamento da estação de pré-tratamento das águas lixiviantes produzidas no aterro sanitário do ecoparque de Palmela, tendo em consideração o facto de este tratamento ser realizado em combinação com o tratamento de águas residuais urbanas. A avaliação do funcionamento da estação de pré-tratamento de águas lixiviantes (EPTAL) é realizada de acordo com os dados fornecidos nos relatórios elaborados pela empresa LUSÁGUA, que apresentam os valores das cargas poluentes medidos à entrada e saída da EPTAL, verificando com estes dados se os valores limite de emissão medidos à saída da EPTAL são respeitados. São ainda propostas algumas alterações baseadas em pareceres técnicos que acompanham os relatórios elaborados pela LUSÁGUA e um estudo elaborada pela ECOserviços em Fevereiro de 2010 onde se obtêm eficiências de etapas de tratamentos com base em testes laboratoriais e testes realizados na própria EPTAL.

Biological activity and cellular uptake of [Ru(eta(5)-C5H5)(PPh3)(Me(2)bpy)][CF3SO3] complex

Anticancer activity of the new [Ru(eta(5)-C5H5)(PPh3)(Me(2)bpy)][CF3SO3] (Me(2)bpy = 4,4'-dimethyl-2,2'-bipyridine) complex was evaluated in vitro against several human cancer cell lines, namely A2780, A2780CisR, HT29, MCF7, MDAMB231 and PC3. Remarkably, the IC50 values, placed in the nanomolar and sub-micromolar range, largely exceeded the activity of cisplatin. Binding to human serum albumin, either HSA (human serum albumin) or HSA(faf) (fatty acid-free human serum albumin) does not affect the complex activity. Fluorescence studies revealed that the present ruthenium complex strongly quench the intrinsic fluorescence of albumin. Cell death by the [Ru(eta(5)-C5H5)(PPh3)(Me(2)bpy)][CF3SO3] complex was reduced in the presence of endocytosis modulators and at low temperature, suggesting an energy-dependent mechanism consistent with endocytosis. On the whole, the biological activity evaluated herein suggests that the complex could be a promising anticancer agent. (C) 2013 Elsevier Inc. All rights reserved.

Comparison of inverse planning systems based on biological or physical factors: Pinnacle®, Corvus® and Monaco®

Radiotherapy (RT) is one of the most important approaches in the treatment of cancer and its performance can be improved in three different ways: through the optimization of the dose distribution, by the use of different irradiation techniques or through the study of radiobiological initiatives. The first is purely physical because is related to the physical dose distributiuon. The others are purely radiobiological because they increase the differential effect between the tumour and the health tissues. The Treatment Planning Systems (TPS) are used in RT to create dose distributions with the purpose to maximize the tumoral control and minimize the complications in the healthy tissues. The inverse planning uses dose optimization techniques that satisfy the criteria specified by the user, regarding the target and the organs at risk (OAR’s). The dose optimization is possible through the analysis of dose-volume histograms (DVH) and with the use of computed tomography, magnetic resonance and other digital image techniques.

Cobalt complexes with pyrazole ligands as catalyst precursors for the peroxidative oxidation of cyclohexane: Z-ray absorption spectroscopy studies and biological applications

[CoCl(-Cl)(Hpz(Ph))(3)](2) (1) and [CoCl2(Hpz(Ph))(4)] (2) were obtained by reaction of CoCl2 with HC(pz(Ph))(3) and Hpz(Ph), respectively (Hpz(Ph)=3-phenylpyrazole). The compounds were isolated as air-stable solids and fully characterized by IR and far-IR spectroscopy, MS(ESI+/-), elemental analysis, cyclic voltammetry (CV), controlled potential electrolysis, and single-crystal X-ray diffraction. Electrochemical studies showed that 1 and 2 undergo single-electron irreversible (CoCoIII)-Co-II oxidations and (CoCoI)-Co-II reductions at potentials measured by CV, which also allowed, in the case of dinuclear complex 1, the detection of electronic communication between the Co centers through the chloride bridging ligands. The electrochemical behavior of models of 1 and 2 were also investigated by density functional theory (DFT) methods, which indicated that the vertical oxidation of 1 and 2 (that before structural relaxation) affects mostly the chloride and pyrazolyl ligands, whereas adiabatic oxidation (that after the geometry relaxation) and reduction are mostly metal centered. Compounds 1 and 2 and, for comparative purposes, other related scorpionate and pyrazole cobalt complexes, exhibit catalytic activity for the peroxidative oxidation of cyclohexane to cyclohexanol and cyclohexanone under mild conditions (room temperature, aqueous H2O2). Insitu X-ray absorption spectroscopy studies indicated that the species derived from complexes 1 and 2 during the oxidation of cyclohexane (i.e., Ox-1 and Ox-2, respectively) are analogous and contain a Co-III site. Complex 2 showed low invitro cytotoxicity toward the HCT116 colorectal carcinoma and MCF7 breast adenocarcinoma cell lines.

New water-soluble Ruthenium(II) cytotoxic complex: biological activity and celular distribution

A novel water soluble organometallic compound, [RuCp(mTPPMSNa)(2,2'-bipy)][CF3SO3] (TM85, where Cp=eta(5)-cyclopentadienyl, mTPPMS = diphenylphosphane-benzene-3-sulfonate and 2,2'-bipy = 2,2'-bipyridine) is presented herein. Studies of interactions with relevant proteins were performed to understand the behavior and mode of action of this complex in the biological environment. Electrochemical and fluorescence studies showed that TM85 strongly binds to albumin. Studies carried out to study the formation of TM85 which adducts with ubiquitin and cytochrome c were performed by electrospray ionization mass spectrometry (ESI-MS). Antitumor activity was evaluated against a variety of human cancer cell lines, namely A2780, A2780cisR, MCF7, MDAMB231, HT29, PC3 and V79 non-tumorigenic cells and compared with the reference drug cisplatin. TM85 cytotoxic effect was reduced in the presence of endocytosis modulators at low temperatures, suggesting an energy-dependent mechanism consistent with endocytosis. Ultrastructural analysis by transmission electron microscopy (TEM) revealed that TM85 targets the endomembranar system disrupting the Golgi and also affects the mitochondria. Disruption of plasma membrane observed by flow cytometry could lead to cellular damage and cell death. On the whole, the biological activity evaluated herein combined with the water solubility property suggests that complex TM85 could be a promising anticancer agent. (C) 2013 Elsevier Inc. All rights reserved.

Protein-Polysaccharides of trametes versocolor: production and biological activities

Extracellular-(E-PPS) and intracellular-protein-polysaccharides (I-PPS) complexes were produced by Trametes versicolor in submerged cultures with different carbon sources. The highest extracellular-(EPS) and intracellular-polysaccharide (IPS) concentration in the complexes was obtained with tomato pomace culture. DPPH radical scavenging for E-PPS and I-PPS produced by liter of culture was equivallent to 2.115 +/- A 0.227 and 1.374 +/- A 0.364 g of ascorbic acid, respectively. These complexes showed a protector effect in the oxidation of erythrocyte membranes and had ability to inhibit the hemolysis and methemoglobin synthesis in stressed erythrocytes. These results suggest that extracellular- and intracellular- polysaccharides produced are important bioactive compounds with medicinal potential.

The efficiency of urine dipsticks for the diagnosis of urinary tract infection

Urinary tract infection (UTI) is one of the most prevalent pathologies in developed countries, particularly in women, characterized by the presence of bacterial growth in any part of the urinary system. Currently, urine culture is considered the gold standard method for the diagnosis of UTI. However, this method has several disadvantages including the time necessary for obtaining the results and the associated high costs. Therefore, it is important to evaluate new efficient and valuable methods for the diagnosis of these infections. Objectives: Presently, dipsticks are considered a possible valuable alternative to urine culture. This method has very low costs associated and the results can be obtained in few minutes. Here we aim to compare the sensibility, specificity, predictive value of a positive test and a negative test of both methods in order to determine the efficiency of the test strips method and also to characterize the microorganism more frequently isolated.

Biological activities and oil properties of Origanum glandulosum Desf: a review

Origanum glandulosum Desf. (Species endemic of North Africa: Tunisia and Algeria) is important medicinally as it has antimicrobial, antifungal, antioxidant, antibacterial, antithrombin, antimutagenic, angiogenic, antiparasetic and antihyperglycaemic activities. Phytochemical investigations of the species of this genus have resulted in the extraction of a number of important bioactive compounds. This emphasizes on the need of extensive study for reporting the additional information on the medicinal importance, the biological activities and properties of oil of other unattended species of Origanum glandulosum. © 2015 Springer-Verlag France.