Comparação da fixação em formalina e em fixador molecular e sua influência em técnicas histoquímicas em tecido hepático

A Formalina 10% Tamponada (FNT 10%) é considerada o fixador de eleição nos Laboratórios de Anatomia Patológica. Contudo, a exposição a esta substância acarreta riscos para a saúde dos técnicos. A International Agency for Research on Cancer (IARC) classificou-a como cancerígeno humano e estudos relevam uma correlação positiva entre a exposição a formaldeído e o desenvolvimento de leucemia e leucemia mielóide. Torna-se relevante alterar o processo de fixação substituindo a FNT 10% por outros fixadores melhorando estas questões. Como tal, desenvolveram-se fixadores à base de outros compostos químicos que não formaldeído, como o Fixador Molecular Universal (UMFIX) que tem como base metanol e polietilenoglicol e que é usualmente utilizado com um processador rápido de micro-ondas. Pretende-se comparar as diferenças entre a fixação por FNT 10% e a fixação por UMFIX, em tecido hepático processado em processador rápido de micro-ondas, para as colorações de Hematoxilina-Eosina (H&E), Tricrómio de Gomori, Reticulina e PAS, através da qualidade final das lâminas testadas. A coloração de H&E foi a única que apresentou diferenças estatisticamente significativas entre os dois fixadores (p=0,032; α<0,05; Mann-Whitney). O UMFIX apresenta-se como um substituto da FNT 10% para o processamento rápido em micro-ondas, pois além de apresentar lâminas com uma qualidade final semelhante ou superior às da FNT 10%, ultrapassa os riscos referidos.

Towards the detection of deception in interactive multimedia environments

A classical application of biosignal analysis has been the psychophysiological detection of deception, also known as the polygraph test, which is currently a part of standard practices of law enforcement agencies and several other institutions worldwide. Although its validity is far from gathering consensus, the underlying psychophysiological principles are still an interesting add-on for more informal applications. In this paper we present an experimental off-the-person hardware setup, propose a set of feature extraction criteria and provide a comparison of two classification approaches, targeting the detection of deception in the context of a role-playing interactive multimedia environment. Our work is primarily targeted at recreational use in the context of a science exhibition, where the main goal is to present basic concepts related with knowledge discovery, biosignal analysis and psychophysiology in an educational way, using techniques that are simple enough to be understood by children of different ages. Nonetheless, this setting will also allow us to build a significant data corpus, annotated with ground-truth information, and collected with non-intrusive sensors, enabling more advanced research on the topic. Experimental results have shown interesting findings and provided useful guidelines for future work. Pattern Recognition

Fast growing fungi: a problem to be solved to achieve characterization of occupational exposure to fungi in cork industry

Chrysonilia sitophila is a common mould in cork industry and has been identified as a cause of IgE sensitization and occupational asthma. This fungal species have a fast growth rate that may inhibit others species’ growth causing underestimated data from characterization of occupational fungal exposure. Aiming to ascertain occupational exposure to fungi in cork industry, were analyzed papers from 2000 about the best air sampling method, to obtain quantification and identification of all airborne culturable fungi, besides the ones that have fast-growing rates. Impaction method don’t allows the collection of a representative air volume, because even with some media that restricts the growth of the colonies, in environments with higher fungal load, such as cork industry, the counting of the colonies is very difficult. Otherwise, impinger method permits the collection of a representative air volume, since we can make dilution of the collected volume. Besides culture methods that allows fungal identification trough macro- and micro-morphology, growth features, thermotolerance and ecological data, we can apply molecular biology with the impinger method, to detect the presence of non-viable particles and potential mycotoxin producers’ strains, and also to detect mycotoxins presence with ELISA or HPLC. Selection of the best air sampling method in each setting is crucial to achieve characterization of occupational exposure to fungi. Information about the prevalent fungal species in each setting and also the eventual fungal load it’s needed for a criterious selection.

Molecular epidemiology of Aspergillus collected from cystic fibrosis patients

Background - Aspergillus respiratory infection is a common complication in cystic fibrosis (CF) and is associated with loss of pulmonary function and allergic disease. Methods - Fifty-three Aspergillus isolates recovered from CF patients were identified to species by Internal Transcribed Spacer Region (ITS), β-tubulin, and calmodulin sequencing. Results - Three species complexes (Terrei, Nigri, and Fumigati) were found. Identification to species level gave a single Aspergillus terreus sensu stricto, one Aspergillus niger sensu stricto and 51 Aspergillus fumigatus sensu stricto isolates. No cryptic species were found. Conclusions - To our knowledge, this is the first prospective study of Aspergillus species in CF using molecular methods. The paucity of non-A. fumigatus and of cryptic species of A. fumigatus suggests a special association of A. fumigatus sensu stricto with CF airways, indicating it likely displays unique characteristics making it suitable for chronic residence in that milieu. These findings could refine an epidemiologic and therapeutic approach geared to this pathogen.

Potential pathogenic fungi assessment through molecular biology in cork industry

Portugal has been the world leader in the cork sector in terms of exports, employing ten thousands of workers. In this working activity, the permanent contact with cork may lead to the exposure to fungi, raising concerns as potential occupational hazards in cork industry. The application of molecular tools is crucial in this setting, since fungal species with faster growth rates may hide other species with clinical relevance, such as species belonging to P. glabrum and A. fumigatus complexes. A study was developed aiming at assessing fungal contamination due to Aspergillus fumigatus complex and Penicillium glabrum complex by molecular methods in three cork industries in the outskirt of Lisbon city.

Improving accuracy for OTD based 3G geolocation in real urban/suburban environments

This paper presents the recent research results about the development of a Observed Time Difference (OTD) based geolocation algorithm based on network trace data, for a real Universal Mobile Telecommunication System (UMTS) Network. The initial results have been published in [1], the current paper focus on increasing the sample convergence rate, and introducing a new filtering approach based on a moving average spatial filter, to increase accuracy. Field tests have been carried out for two radio environments (urban and suburban) in the Lisbon area, Portugal. The new enhancements produced a geopositioning success rate of 47% and 31%, and a median accuracy of 151 m and 337 m, for the urban and suburban environments, respectively. The implemented filter produced a 16% and 20% increase on accuracy, when compared with the geopositioned raw data. The obtained results are rather promising in accuracy and geolocation success rate. OTD positioning smoothed by moving average spatial filtering reveals a strong approach for positioning trace extracted events, vital for boosting Self-Organizing Networks (SON) over a 3G network.

Virtual laboratory for educational environments

In this paper a new simulation environment for a virtual laboratory to educational proposes is presented. The Logisim platform was adopted as the base digital simulation tool, since it has a modular implementation in Java. All the hardware devices used in the laboratory course was designed as components accessible by the simulation tool, and integrated as a library. Moreover, this new library allows the user to access an external interface. This work was motivated by the needed to achieve better learning times on co-design projects, based on hardware and software implementations, and to reduce the laboratory time, decreasing the operational costs of engineer teaching. Furthermore, the use of virtual laboratories in educational environments allows the students to perform functional tests, before they went to a real laboratory. Moreover, these functional tests allow to speed-up the learning when a problem based approach methodology is considered. © 2014 IEEE.

Molecular mobility, composition and structure analysis in glycerol plasticised chitosan films

This study was developed with the purpose to investigate the effect of polysaccharide/plasticiser concentration on the microstructure and molecular dynamics of polymeric film systems, using transmission electron microscope imaging (TEM) and nuclear magnetic resonance (NMR) techniques. Experiments were carried out in chitosan/glycerol films prepared with solutions of different composition. The films obtained after drying and equilibration were characterised in terms of composition, thickness and water activity. Results show that glycerol quantities used in film forming solutions were responsible for films composition; while polymer/total plasticiser ratio in the solution determined the thickness (and thus structure) of the films. These results were confirmed by TEM. NMR allowed understanding the films molecular rearrangement. Two different behaviours for the two components analysed, water and glycerol were observed: the first is predominantly moving free in the matrix, while glycerol is mainly bounded to the chitosan chain. (C) 2013 Elsevier Ltd. All rights reserved.

2D PCA-Based localization for mobile robots in unstructured environments

In this paper a new PCA-based positioning sensor and localization system for mobile robots to operate in unstructured environments (e. g. industry, services, domestic ...) is proposed and experimentally validated. The inexpensive positioning system resorts to principal component analysis (PCA) of images acquired by a video camera installed onboard, looking upwards to the ceiling. This solution has the advantage of avoiding the need of selecting and extracting features. The principal components of the acquired images are compared with previously registered images, stored in a reduced onboard image database, and the position measured is fused with odometry data. The optimal estimates of position and slippage are provided by Kalman filters, with global stable error dynamics. The experimental validation reported in this work focuses on the results of a set of experiments carried out in a real environment, where the robot travels along a lawn-mower trajectory. A small position error estimate with bounded co-variance was always observed, for arbitrarily long experiments, and slippage was estimated accurately in real time.

On the assessment of exposure to airborne ultrafine particles in urban environments

The aim of this study was to contribute to the assessment of exposure levels of ultrafine particles in the urban environment of Lisbon, Portugal, due to automobile traffic, by monitoring lung deposited alveolar surface area (resulting from exposure to ultrafine particles) in a major avenue leading to the town center during late spring, as well as in indoor buildings facing it. Data revealed differentiated patterns for week days and weekends, consistent with PM2.5 and PM10 patterns currently monitored by air quality stations in Lisbon. The observed ultrafine particulate levels may be directly correlated with fluxes in automobile traffic. During a typical week, amounts of ultrafine particles per alveolar deposited surface area varied between 35 and 89.2 mu m2/cm3, which are comparable with levels reported for other towns in Germany and the United States. The measured values allowed for determination of the number of ultrafine particles per cubic centimeter, which are comparable to levels reported for Madrid and Brisbane. In what concerns outdoor/indoor levels, we observed higher levels (32 to 63%) outdoors, which is somewhat lower than levels observed in houses in Ontario.

Lighting conditions and optical filters effects on visual performance of speleologists exposed to cave environments

The aim of this study is to evaluate lighting conditions and speleologists’ visual performance using optical filters when exposed to the lighting conditions of cave environments. A crosssectional study was conducted. Twenty-three speleologists were submitted to an evaluation of visual function in a clinical lab. An examination of visual acuity, contrast sensitivity, stereoacuity and flashlight illuminance levels was also performed in 16 of the 23 speleologists at two caves deprived of natural lightning. Two organic filters (450 nm and 550 nm) were used to compare visual function with and without filters. The mean age of the speleologists was 40.65 (± 10.93) years. We detected 26.1% participants with visual impairment of which refractive error (17.4%) was the major cause. In the cave environment the majority of the speleologists used a head flashlight with a mean illuminance of 451.0 ± 305.7 lux. Binocular visual acuity (BVA) was -0.05 ± 0.15 LogMAR (20/18). BVA for distance without filter was not statistically different from BVA with 550 nm or 450 nm filters (p = 0.093). Significant improved contrast sensitivity was observed with 450 nm filters for 6 cpd (p = 0.034) and 18 cpd (p = 0.026) spatial frequencies. There were no signs and symptoms of visual pathologies related to cave exposure. Illuminance levels were adequate to the majority of the activities performed. The enhancement in contrast sensitivity with filters could potentially improve tasks related with the activities performed in the cave.

A cellulosic liquid crystal pool for cellulose nanocrystals: structure and molecular dynamics at high shear rates

Cellulose and its derivatives, such as hydroxypropylcellulose (HPC) have been studied for a long time but they are still not well understood particularly in liquid crystalline solutions. These systems can be at the origin of networks with properties similar to liquid crystalline (LC) elastomers. The films produced from LC solutions can be manipulated by the action of moisture allowing for instance the development of a soft motor (Geng et al., 2013) driven by humidity. Cellulose nanocrystals (CNC), which combine cellulose properties with the specific characteristics of nanoscale materials, have been mainly studied for their potential as a reinforcing agent. Suspensions of CNC can also self-order originating a liquid-crystalline chiral nematic phases. Considering the liquid crystalline features that both LC-HPC and CNC can acquire, we prepared LC-HPC/CNC solutions with different CNC contents (1,2 and 5 wt.%). The effect of the CNC into the LC-HPC matrix was determined by coupling rheology and NMR spectroscopy - Rheo-NMR a technique tailored to analyse orientational order in sheared systems. (C) 2015 Elsevier Ltd. All rights reserved.

Molecular fingerprint of human gastric cell line infected by Helicobacter pylori

Helicobacter pylori infection represents a serious health problem, given its association with serious gastric diseases as gastric ulcers, cancer and MALT lymphoma. Currently no vaccine exists and antibiotic-based eradication therapy is already failing in more than 20% of cases. To increase the knowledge on the infection process diverse gastric cell lines, e.g. the adenocarcinona gastric (AGS) cell line, are routinely used has in vitro models of gastric epithelia. In the present work the molecular fingerprint of infected and non-infected AGS cell lines, by diverse H. pylori strains, was acquired using vibrational infrared spectroscopy. These molecular fingerprints enabled to discriminate infected from non-infected AGS cells, and infection due to different strains, by performing Principal Component Analysis. It was also possible to estimate, from the AGS cells molecular fingerprint, the effect of the infection on diverse biochemical and metabolic cellular status. In resume infra-red spectroscopy enabled the acquisition of infected AGS cells molecular fingerprint with minimal sample preparation, in a rapid, high-throughput, economic process yielding highly sensitive and informative data, most useful for promoting critical knowledge on the H. pylori infection process. © 2015 IEEE.

Molecular details of INH-C-10 binding to wt KatG and Its S315T mutant

Isoniazid (INH) is still one of the two most effective antitubercular drugs and is included in all recommended multitherapeutic regimens. Because of the increasing resistance of Mycobacterium tuberculosis to INH, mainly associated with mutations in the katG gene, new INH-based compounds have been proposed to circumvent this problem. In this work, we present a detailed comparative study of the molecular determinants of the interactions between wt KatG or its S315T mutant form and either INH or INH-C10, a new acylated INH derivative. MD simulations were used to explore the conformational space of both proteins, and results indicate that the S315T mutation did not have a significant impact on the average size of the access tunnel in the vicinity of these residues. Our simulations also indicate that the steric hindrance role assigned to Asp137 is transient and that electrostatic changes can be important in understanding the enzyme activity data of mutations in KatG. Additionally, molecular docking studies were used to determine the preferred modes of binding of the two substrates. Upon mutation, the apparently less favored docking solution for reaction became the most abundant, suggesting that S315T mutation favors less optimal binding modes. Moreover, the aliphatic tail in INH-C10 seems to bring the hydrazine group closer to the heme, thus favoring the apparent most reactive binding mode, regardless of the enzyme form. The ITC data is in agreement with our interpretation of the C10 alkyl chain role and helped to rationalize the significantly lower experimental MIC value observed for INH-C10. This compound seems to be able to counterbalance most of the conformational restrictions introduced by the mutation, which are thought to be responsible for the decrease in INH activity in the mutated strain. Therefore, INH-C10 appears to be a very promising lead compound for drug development.