Fungi spores dimension matters in health effects: a methodology for more detail fungi exposure assessment

Health effects resulting from dust inhalation in occupational environments may be more strongly associated with specific microbial components, such as fungi, than to the particles. The aim of the present study is to characterize the occupational exposure to the fungal burden in four different occupational settings (two feed industries, one poultry and one waste sorting industry), presenting results from two air sampling methods – the impinger collector and the use of filters. In addition, the equipment used for the filter sampling method allowed a more accurate characterization regarding the dimension of the collected fungal particles (less than 2.5 μm size). Air samples of 300L were collected using the impinger Coriolis μ air sampler. Simultaneously, the aerosol monitor (DustTrak II model 8532, TSI®) allowed assessing viable microbiological material below the 2.5 μm size. After sampling, filters were immersed in 300 mL of sterilized distilled water and agitated for 30 min at 100 rpm. 150 μl from the sterilized distilled water were subsequently spread onto malt extract agar (2%) with chloramphenicol (0.05 g/L). All plates were incubated at 27.5 ºC during 5–7 days. With the impinger method, the fungal load ranged from 0 to 413 CFU.m-3 and with the filter method, ranged from 0 to 64 CFU.m-3. In one feed industry, Penicillium genus was the most frequently found genus (66.7%) using the impinger method and three more fungi species/genera/complex were found. The filter assay allowed the detection of only two species/genera/complex in the same industry. In the other feed industry, Cladosporium sp. was the most found (33.3%) with impinger method and three more species/genera/complex were also found. Through the filter assay four fungi species/genera/complex were found. In the assessed poultry, Rhyzopus sp. was the most frequently detected (61.2%) and more three species/genera/complex were isolated. Through the filter assay, only two fungal species/genera/complex were found. In the waste sorting industry Penicillium sp. was the most prevalent (73.6%) with the impinger method, being isolated two more different fungi species/genera/complex. Through the filter assay only Penicillium sp. was found. A more precise determination of occupational fungal exposure was ensured, since it was possible to obtain information regarding not only the characterization of fungal contamination (impinger method), but also the size of dust particles, and viable fungal particles, that can reach the worker ́s respiratory tract (filters method). Both methods should be used in parallel to enrich discussion regarding potential health effects of occupational exposure to fungi.

Preoperative assessment of deep myometrial and cervical invasion in endometrial carcinoma: comparison of magnetic resonance imaging and histopathologic evaluation

This study aimed to evaluate the accuracy of magnetic resonance imaging (MRI) in the detection of deep myometrial invasion and cervical extension by endometrial carcinoma. This prospective study included 101 patients with histologically documented endometrial carcinoma, between July 1998 and April 2004. The findings of preoperative pelvic MRI were compared with histological diagnosis. From 101 cases studied by pelvic MRI, 43 were classified as deep myometrial invasion (50% of myometrium), where the pathological evaluation confirmed as having deep myometrial invasion. Cervical extension in the MRI study was found in 19 cases. Pathologic study found cervical extension and/or invasion in 31 cases including all cases identified by MRI. The accuracy, sensitivity and specificity of MRI were 95%, 89%, 100%, detecting deep myometrial invasion and 88%, 61%, 100%, detecting cervical invasion, respectively. The high accuracy achieved makes MRI an adequate method for determine the depth of myometrial and cervical invasion in endometrial carcinoma.

Domestic Rhodnius ecuadoriensis (Hemiptera, Reduviidae) infestation in Northern Peru: a comparative trial of detection methods during a six-month follow-up

Two passive methods in the assessment of intradomiciliary infestation by Rhodnius ecuadoriensis were tested: (i) the Gomes Nuñez sensor box (GN), (ii) sheets of white typing paper and (iii) one active timed manual method. The study was carried out in the Alto Chicama River Valley, Province of Gran Chimú, Department of La Libertad. The study design consisted of an initial searching of triatomines inside of the domestic environment by the manual capture active procedure (man/hour) covering all the studied houses. Then, matched pairs of GN boxes and paper sheets were simultaneously installed in the bedrooms of 207 households distributed in 19 localities. A comparative prospective trial of these passive detection devices were monitored at 2, 4 and, finally 6 months follow-up. Parasitological Trypanosoma rangeli and/or T. cruzi infections were investigated in two houses with high level of infestation by R. ecuadoriensis.16.9% of the 207 households investigated by an initial active manual method were infested with R. ecuadoriensis. The proportion of infested houses fluctuated from 6.2 to 55.5% amongst the 19 localities investigated. T. rangeli natural infection was detected in R.ecuadoriensis specimens collected in two households. Parasite rates in the bugs ranged from 16.6 to 21.7% respectively. The most striking fact was an average rate of salivary gland infection ranging from 7.4 to 8.3%. At the end of the sixth month period, a cumulative incidence of 31.4% of positive GN boxes against 15.9% for paper sheets was recorded. All three methods combined detected domestic infestation in 129 (62.3%) of the 207 houses studied in the 19 localities. The range of houses infested varies from 6.7% to 92.9%. In areas with low bug density infestation rates, the methodology experienced in our studies, seems to be the best choice for investigations on domestic R. ecuadoriensis populations.

Adaptação de um instrumento de rastreio cognitivo, o MoCA (Montreal Cognitive Assessment) para população cabo-verdiana

Dissertação de Mestrado, Neurociências Cognitivas e Neuropsicologia, Faculdade de Ciências Humanas e Sociais, Universidade do Algarve, 2016

IMPLEMENTATION OF PHOTOGRAMMETRY TO IMPROVE PROACTIVE ASSESSMENT OF RETAINING WALLS ALONG TRANSPORTATION CORRIDORS

Retaining walls are important assets in the transportation infrastructure and assessing their condition is important to prolong their performance and ultimately their design life. Retaining walls are often overlooked and only a few transportation asset management programs consider them in their inventory. Because these programs are few, the techniques used to assess their condition focus on a qualitative assessment as opposed to a quantitative approach. The work presented in this thesis focuses on using photogrammetry to quantitatively assess the condition of retaining walls. Multitemporal photogrammetry is used to develop 3D models of the retaining walls, from which offset displacements are measured to assess their condition. This study presents a case study from a site along M-10 highway in Detroit, MI were several sections of retaining walls have experienced horizontal displacement towards the highway. The results are validated by comparing with field observations and measurements. The limitations of photogrammetry were also studied by using a small scale model in the laboratory. The analysis found that the accuracy of the offset displacement measurements is dependent on the distance between the retaining wall and the sensor, location of the reference points in 3D space, and the focal length of the lenses used by the camera. These parameters were not ideal for the case study at the M-10 highway site, but the results provided consistent trends in the movement of the retaining wall that couldn’t be validated from offset measurements. The findings of this study confirm that photogrammetry shows promise in generating 3D models to provide a quantitative condition assessment for retaining walls within its limitations.

A measure of skin familiarity and its role in the early detection of skin cancer

Skin self-examination (SSE) is promoted widely so that individuals will become familiar with their skin and be better able to identify suspicious changes earlier. However, individuals can also become familiar with their skin other than through purposeful SSE. In this article, we develop a measure of skin familiarity based on the density of spots on 14 different areas of the body. A factor analysis of the 14 body-area scores revealed that they could be grouped into four broad body regions (shoulders and back, front of legs, back of legs, and feet). Each total body score and body-region score has high internal consistency (Cronbach's alpha coefficients ranging from 0.79 to 0.93). Moreover, the scores correlate as expected with skin self-examination behaviors and other personal characteristics, indicating high construct validity. We consider the advantages that skin familiarity measures offer over the exclusive use of SSE measures in the assessment of early detection activities and discuss the direction of future research in this area

Assessment of the Occurrence and Potential Risks of Antibiotics and their Metabolites in South Florida Waters Using Liquid Chromatography Tandem Mass Spectrometry

An automated on-line SPE-LC-MS/MS method was developed for the quantitation of multiple classes of antibiotics in environmental waters. High sensitivity in the low ng/L range was accomplished by using large volume injections with 10-mL of sample. Positive confirmation of analytes was achieved using two selected reaction monitoring (SRM) transitions per antibiotic and quantitation was performed using an internal standard approach. Samples were extracted using online solid phase extraction, then using column switching technique; extracted samples were immediately passed through liquid chromatography and analyzed by tandem mass spectrometry. The total run time per each sample was 20 min. The statistically calculated method detection limits for various environmental samples were between 1.2 and 63 ng/L. Furthermore, the method was validated in terms of precision, accuracy and linearity. The developed analytical methodology was used to measure the occurrence of antibiotics in reclaimed waters (n=56), surface waters (n=53), ground waters (n=8) and drinking waters (n=54) collected from different parts of South Florida. In reclaimed waters, the most frequently detected antibiotics were nalidixic acid, erythromycin, clarithromycin, azithromycin trimethoprim, sulfamethoxazole and ofloxacin (19.3-604.9 ng/L). Detection of antibiotics in reclaimed waters indicates that they can’t be completely removed by conventional wastewater treatment process. Furthermore, the average mass loads of antibiotics released into the local environment through reclaimed water were estimated as 0.248 Kg/day. Among the surface waters samples, Miami River (reaching up to 580 ng/L) and Black Creek canal (up to 124 ng/L) showed highest concentrations of antibiotics. No traces of antibiotics were found in ground waters. On the other hand, erythromycin (monitored as anhydro erythromycin) was detected in 82% of the drinking water samples (n.d-66 ng/L). The developed approach is suitable for both research and monitoring applications. Major metabolites of antibiotics in reclaimed wates were identified and quantified using high resolution benchtop Q-Exactive orbitrap mass spectrometer. A phase I metabolite of erythromycin was tentatively identified in full scan based on accurate mass measurement. Using extracted ion chromatogram (XIC), high resolution data-dependent MS/MS spectra and metabolic profiling software the metabolite was identified as desmethyl anhydro erythromycin with molecular formula C36H63NO12 and m/z 702.4423. The molar concentration of the metabolite to erythromycin was in the order of 13 %. To my knowledge, this is the first known report on this metabolite in reclaimed water. Another compound acetyl-sulfamethoxazole, a phase II metabolite of sulfamethoxazole was also identified in reclaimed water and mole fraction of the metabolite represent 36 %, of the cumulative sulfamethoxazole concentration. The results were illustrating the importance to include metabolites also in the routine analysis to obtain a mass balance for better understanding of the occurrence, fate and distribution of antibiotics in the environment. Finally, all the antibiotics detected in reclaimed and surface waters were investigated to assess the potential risk to the aquatic organisms. The surface water antibiotic concentrations that represented the real time exposure conditions revealed that the macrolide antibiotics, erythromycin, clarithromycin and tylosin along with quinolone antibiotic, ciprofloxacin were suspected to induce high toxicity to aquatic biota. Preliminary results showing that, among the antibiotic groups tested, macrolides posed the highest ecological threat, and therefore, they may need to be further evaluated with, long-term exposure studies considering bioaccumulation factors and more number of species selected. Overall, the occurrence of antibiotics in aquatic environment is posing an ecological health concern.

Real-time Biosensor for the Assessment of Nanotoxicity and Cancer Electrotherapy

Knowledge of cell electronics has led to their integration to medicine either by physically interfacing electronic devices with biological systems or by using electronics for both detection and characterization of biological materials. In this dissertation, an electrical impedance sensor (EIS) was used to measure the electrode surface impedance changes from cell samples of human and environmental toxicity of nanoscale materials in 2D and 3D cell culture models. The impedimetric response of human lung fibroblasts and rainbow trout gill epithelial cells when exposed to various nanomaterials was tested to determine their kinetic effects towards the cells and to demonstrate the biosensor’s ability to monitor nanotoxicity in real-time. Further, the EIS allowed rapid, real-time and multi-sample analysis creating a versatile, noninvasive tool that is able to provide quantitative information with respect to alteration in cellular function. We then extended the application of the unique capabilities of the EIS to do real-time analysis of cancer cell response to externally applied alternating electric fields at different intermediate frequencies and low-intensity. Decreases in the growth profiles of the ovarian and breast cancer cells were observed with the application of 200 and 100 kHz, respectively, indicating specific inhibitory effects on dividing cells in culture in contrast to the non-cancerous HUVECs and mammary epithelial cells. We then sought to enhance the effects of the electric field by altering the cancer cell’s electronegative membrane properties with HER2 antibody functionalized nanoparticles. An Annexin V/EthD-III assay and zeta potential were performed to determine the cell death mechanism indicating apoptosis and a decrease in zeta potential with the incorporation of the nanoparticles. With more negatively charged HER2-AuNPs attached to the cancer cell membrane, the decrease in membrane potential would thus leave the cells more vulnerable to the detrimental effects of the applied electric field due to the decrease in surface charge. Therefore, by altering the cell membrane potential, one could possibly control the fate of the cell. This whole cell-based biosensor will enhance our understanding of the responsiveness of cancer cells to electric field therapy and demonstrate potential therapeutic opportunities for electric field therapy in the treatment of cancer.

Long-term assessment of wild boar harvesting and cattle removal for bovine tuberculosis control in free ranging populations

Wild boar is a recognized reservoir of bovine tuberculosis (TB) in the Mediterranean ecosystems, but information is scarce outside of hotspots in southern Spain. We describe the first high-prevalence focus of TB in a non-managed wild boar population in northern Spain and the result of eight years of TB management. Measures implemented for disease control included the control of the local wild boar population through culling and stamping out of a sympatric infected cattle herd. Post-mortem inspection for detection of tuberculosis-like lesions as well as cultures from selected head and cervical lymph nodes was done in 745 wild boar, 355 Iberian ibexes and five cattle between 2004 and 2012. The seasonal prevalence of TB reached 70% amongst adult wild boar and ten different spoligotypes and 13 MIRU-VNTR profiles were detected, although more than half of the isolates were included in the same clonal complex. Only 11% of infected boars had generalized lesions. None of the ibexes were affected, supporting their irrelevance in the epidemiology of TB. An infected cattle herd grazed the zone where 168 of the 197 infected boars were harvested. Cattle removal and wild boar culling together contributed to a decrease in TB prevalence. The need for holistic, sustained over time, intensive and adapted TB control strategies taking into account the multi-host nature of the disease is highlighted. The potential risk for tuberculosis emergence in wildlife scenarios where the risk is assumed to be low should be addressed.

Early stage assessment of the inspectability of welded components : A case from the aerospace industry

This paper proposes a method to indicate potential problems when planning dye penetrant and x-ray inspection of welded components. Inspection has been found to be an important part of the manufacturability evaluation made in a large CAD-based parametric environment for making multidisciplinary design simulations in early stages of design at an aircraft component manufacturer. The paper explains how the proposed method is to be included in the design platform at the company. It predicts the expected probability of detection of cracks (POD) in situations where the geometry of the parts is unfavourable for inspection so that potential problems can be discovered and solved in early stages. It is based on automatically extracting information from CAD-models and making a rule-based evaluation. It also provides a scale for how favourable the geometry is for inspection. In the paper it is also shown that the manufacturability evaluation need to take into consideration the expected stresses in the structures, highlighting the importance of multi-disciplinary simulations.

Experimentation of mitigation techniques to reduce the effects of permafrost degradation on transportation infrastructures at Beaver Creek experimental road site, Alaska Highway, Yukon

Les méthodes de design et de construction des routes développés dans le sud canadien ont maintenant besoin d’être adaptés aux environnements nordiques du pays afin de prévenir le dégel dramatique du pergélisol lors de la construction d’une nouvelle route. De plus, le réchauffement climatique occasionne présentement d’importants problèmes de stabilité des sols dans le nord canadien. Ces facteurs causent des pertes importantes au niveau des capacités fonctionnelles et structurales de l’Alaska Highway au Yukon sur un segment de plus de 200 km situé entre le village de Destruction Bay et la frontière de l’Alaska. Afin de trouver des solutions rentables à long terme, le ministère du transport du Yukon (en collaboration avec le Federal Highway Administration du gouvernement américain, Transports Canada, l’Université Laval, l’Université de Montréal et l’Alaska University transportation Center) a mis en place 12 sections d’essais de 50 mètres de longueur sur l’autoroute de l’Alaska près de Beaver Creek en 2008. Ces différentes sections d’essais ont été conçues pour évaluer une ou plusieurs méthodes combinées de stabilisation thermique telles que le drain thermique, le remblai à convection d’air, le pare-neige / pare-soleil, le remblai couvert de matières organiques, les drains longitudinaux, le déblaiement de la neige sur les pentes et la surface réfléchissante. Les objectifs spécifiques de la recherche sont 1) d’établir les régimes thermiques et les flux de chaleur dans chacune des sections pour les 3 premières années de fonctionnement ; 2) de documenter les facteurs pouvant favoriser ou nuire à l’efficacité des systèmes de protection et ; 3) de déterminer le rapport coûts/bénéfices à long terme pour chacune des techniques utilisées. Pour ce faire, une nouvelle méthode d’analyse, basée sur la mesure de flux d’extraction de chaleur Hx et d’induction Hi à l’interface entre le remblai et le sol naturel, a été utilisée dans cette étude. Certaines techniques de protection du pergélisol démontrent un bon potentiel durant leurs 3 premières années de fonctionnement. C’est le cas pour le remblai à convection d’air non-couvert, le remblai à convection d’air pleine largeur, les drains longitudinaux, le pare-soleil / pare-neige et la surface réfléchissante. Malheureusement, des problèmes dans l’installation des drains thermiques ont empêché une évaluation complète de leur efficacité.

An investigation into the utility of the Mini Addenbrooke's Cognitive Examination (M-ACE) for the early detection of dementia and mild cognitive impairment in people aged 75 and over

Background/Aims: The Mini Addenbrooke’s Cognitive Examination (M-ACE) is the abbreviated version of the widely-used Addenbrooke’s Cognitive Examination (ACE-III), a cognitive screening tool that is used internationally in the assessment of mild cognitive impairment (MCI) and dementia. The objectives of this study were to investigate the diagnostic accuracy of the M-ACE with individuals aged 75 and over to distinguish between those who do and do not have a dementia or MCI, and also to establish whether the cut-off scores recommended by Hsieh et al. (2014) [9] in the original validation study for the M-ACE are optimal for this age group. Methods: The M-ACE was administered to 58 participants (24 with a diagnosis of dementia, 17 with a diagnosis of MCI and 17 healthy controls). The extent to which scores distinguished between groups (dementia, MCI or no diagnosis) was explored using receiver operating characteristic curve analysis. Results: The optimal cut-off for detecting dementia was ≤ 21/30 (score ≤ 21/30 indicating dementia with a sensitivity of 0.95, a specificity of 1 and a positive predictive value of 1) compared to the original higher published cut-off of ≤ 25/30 (sensitivity of 0.95, specificity of 0.70 and a positive predictive value of 0.82 in this sample). Conclusions: The M-ACE has excellent diagnostic accuracy for the detection of dementia in a UK clinical sample. It may be necessary to consider lower cut-offs than those given in the original validation study.

A micro-extraction technique using a new digitally controlled syringe combined with UHPLC for assessment of urinary biomarkers of oxidatively damaged DNA

The formation of reactive oxygen species (ROS) within cells causes damage to biomolecules, including membrane lipids, DNA, proteins and sugars. An important type of oxidative damage is DNA base hydroxylation which leads to the formation of 8-oxo-7,8-dihydro-29-deoxyguanosine (8-oxodG) and 5-hydroxymethyluracil (5-HMUra). Measurement of these biomarkers in urine is challenging, due to the low levels of the analytes and the matrix complexity. In order to simultaneously quantify 8-oxodG and 5-HMUra in human urine, a new, reliable and powerful strategy was optimised and validated. It is based on a semi-automatic microextraction by packed sorbent (MEPS) technique, using a new digitally controlled syringe (eVolH), to enhance the extraction efficiency of the target metabolites, followed by a fast and sensitive ultrahigh pressure liquid chromatography (UHPLC). The optimal methodological conditions involve loading of 250 mL urine sample (1:10 dilution) through a C8 sorbent in a MEPS syringe placed in the semi-automatic eVolH syringe followed by elution using 90 mL of 20% methanol in 0.01% formic acid solution. The obtained extract is directly analysed in the UHPLC system using a binary mobile phase composed of aqueous 0.1% formic acid and methanol in the isocratic elution mode (3.5 min total analysis time). The method was validated in terms of selectivity, linearity, limit of detection (LOD), limit of quantification (LOQ), extraction yield, accuracy, precision and matrix effect. Satisfactory results were obtained in terms of linearity (r2 . 0.991) within the established concentration range. The LOD varied from 0.00005 to 0.04 mg mL21 and the LOQ from 0.00023 to 0.13 mg mL21. The extraction yields were between 80.1 and 82.2 %, while inter-day precision (n=3 days) varied between 4.9 and 7.7 % and intra-day precision between 1.0 and 8.3 %. This approach presents as main advantages the ability to easily collect and store urine samples for further processing and the high sensitivity, reproducibility, and robustness of eVolHMEPS combined with UHPLC analysis, thus retrieving a fast and reliable assessment of oxidatively damaged DNA.

Screening a Case Base for Stroke Disease Detection

Stroke stands for one of the most frequent causes of death, without distinguishing age or genders. Despite representing an expressive mortality fig-ure, the disease also causes long-term disabilities with a huge recovery time, which goes in parallel with costs. However, stroke and health diseases may also be prevented considering illness evidence. Therefore, the present work will start with the development of a decision support system to assess stroke risk, centered on a formal framework based on Logic Programming for knowledge rep-resentation and reasoning, complemented with a Case Based Reasoning (CBR) approach to computing. Indeed, and in order to target practically the CBR cycle, a normalization and an optimization phases were introduced, and clustering methods were used, then reducing the search space and enhancing the cases re-trieval one. On the other hand, and aiming at an improvement of the CBR theo-retical basis, the predicates` attributes were normalized to the interval 0…1, and the extensions of the predicates that match the universe of discourse were re-written, and set not only in terms of an evaluation of its Quality-of-Information (QoI), but also in terms of an assessment of a Degree-of-Confidence (DoC), a measure of one`s confidence that they fit into a given interval, taking into account their domains, i.e., each predicate attribute will be given in terms of a pair (QoI, DoC), a simple and elegant way to represent data or knowledge of the type incomplete, self-contradictory, or even unknown.

Advanced neuroimaging methodologies to improve connectivity detection in normal and abnormal language brain networks

The language connectome was in-vivo investigated using multimodal non-invasive quantitative MRI. In PPA patients (n=18) recruited by the IRCCS ISNB, Bologna, cortical thickness measures showed a predominant reduction on the left hemisphere (p<0.005) with respect to matched healthy controls (HC) (n=18), and an accuracy of 86.1% in discrimination from Alzheimer’s disease patients (n=18). The left temporal and para-hippocampal gyri significantly correlated (p<0.01) with language fluency. In PPA patients (n=31) recruited by the Northwestern University Chicago, DTI measures were longitudinally evaluated (2-years follow-up) under the supervision of Prof. M. Catani, King’s College London. Significant differences with matched HC (n=27) were found, tract-localized at baseline and widespread in the follow-up. Language assessment scores correlated with arcuate (AF) and uncinate (UF) fasciculi DTI measures. In left-ischemic stroke patients (n=16) recruited by the NatBrainLab, King’s College London, language recovery was longitudinally evaluated (6-months follow-up). Using arterial spin labelling imaging a significant correlation (p<0.01) between language recovery and cerebral blood flow asymmetry, was found in the middle cerebral artery perfusion, towards the right. In HC (n=29) recruited by the DIBINEM Functional MR Unit, University of Bologna, an along-tract algorithm was developed suitable for different tractography methods, using the Laplacian operator. A higher left superior temporal gyrus and precentral operculum AF connectivity was found (Talozzi L et al., 2018), and lateralized UF projections towards the left dorsal orbital cortex. In HC (n=50) recruited in the Human Connectome Project, a new tractography-driven approach was developed for left association fibres, using a principal component analysis. The first component discriminated cortical areas typically connected by the AF, suggesting a good discrimination of cortical areas sharing a similar connectivity pattern. The evaluation of morphological, microstructural and metabolic measures could be used as in-vivo biomarkers to monitor language impairment related to neurodegeneration or as surrogate of cognitive rehabilitation/interventional treatment efficacy.