Development of an in situ penetration test for the uptake of preservatives in applied wood

This paper concerns the study of biocides application in old timber structures of maritime pine (Pinus pinaster Ail.), previously impregnated with other products. A method was developed in laboratory to determine in situ the penetration depth of a product applied superficially. As initial treatment, three traditional products for sawn timber for buildings were used and, for new treatments, two newer, more environmentally benign products were used. Their ability to penetrate the pre-treated surfaces was evaluated after 1, 2 and 3 applications at 24 hours intervals and the results obtained are presented. Finally, the applicability of the developed test to the in-situ evaluation of timber structures is also discussed.

Fungal contamination of poultries litter: a public health problem

Exposure to certain fungi can cause human illness. Fungi cause adverse human health effects through three specific mechanisms: generation of a harmful immune response (e.g., allergy or hypersensitivity pneumonitis); direct infection by the fungal organism; by toxic-irritant effects from mold byproducts, such as mycotoxins. In Portugal there is an increasingly industry of large facilities that produce whole chickens for domestic consumption and only few investigations have reported on fungal contamination of the poultry litter. The material used for poultry litter is varied but normally can be constitute by: pine shavings; sawdust of eucalyptus; other types of wood; peanut; coffee; sugar cane; straw; hay; grass; paper processed. Litter is one of the most contributive factors to fungal contamination in poultries. Spreading litter is one of the tasks that normally involve higher exposure of the poultry workers to dust, fungi and their metabolites, such as VOC’s and mycotoxins. After being used and removed from poultries, litter is ploughed into agricultural soils, being this practice potentially dangerous for the soil environment, as well for both humans and animals. The goal of this study was to characterize litter’s fungal contamination and also to report the incidence of keratinophilic and toxigenic fungi.

Dissecting the role of CLASP1 in the mammalian development

A mitose é o evento celular, através do qual uma células transmite uma cópias do seu DNA às células filhas. Este processo é mediado pelo fuso mitótico, o qual consiste numa rede bipolar microtubulos. A dinâmica dos microtubulos é regulada por proteínas associadas a estes (MAPs – Microtubule-Associated Proteins), tais como as proteínas associadas às extremidades positivas dos microtubulos (+TIPs – Plus-ends Tracking proteins). As proteínas associadas às CLIPs (CLASPs – CLIP-associated proteins) pertencem a esta família e estão altamente conservadas nos eucariotas. Estas interagem com os microtubulos regulando o fuso mitótico, a segregação dos cromossomas e o comportamento dos microtubulos ao nível do cinetocoro. Assim, as CLASPs têm sido descritas como essenciais à manutenção da integridade genética durante a divisão celular. Um modelo animal knockout para o gene Clasp1 é uma ferramenta indispensável à descoberta do papel da CLASP1 a nível fisiológico. Nos animais knockout foi observado um fenótipo letal, no qual 100% dos recém-nascidos morreram poucos minutos após o nascimento, no decurso de falência respiratória. Após análise histopatológica, observamos que os pulmões dos animais knockout apresentam um atraso no desenvolvimento. Porém, a análise da expressão de marcadores de diferenciação celular, mostrou que os pneumócitos tipo I e II estão presente e diferenciados nos animais knockout aquando do seu nascimento. No entanto, um defeito primário a nível pulmonar ainda não pode ser excluído. Níveis elevados de glicogénio no parênquima alveolar dos animais knockout sugerem imaturidade pulmonar ou deficiente produção do líquido surfactante. Adicionalmente, ainda não está esclarecido de que forma pode este atraso explicar a letalidade observada nos recémnascidos knockout. Verificamos também que expressão de CLASP1 é transiente ao longo do desenvolvimento, sendo particularmente elevada no cérebro, o que pode explicar o seu papel já descrito na biologia dos neurónios. A CLASP1 é ubiquamente expressa em mamíferos adultos, o que sugere que esta proteína é também importante em tecidos diferenciados. Nesta fase, o significado biológico da CLASP1 em mamíferos ainda não foi descortinado. No entanto, nenhum animal knockout para Clasp1 foi capaz de sobreviver ex uterus, o que sugere um papel fundamental desta proteína na fase final do desenvolvimento dos mamíferos.

Fungal and microbial volatile organic compounds exposure assessment in a waste sorting plant

In the management of solid waste, pollutants over a wide range are released with different routes of exposure for workers. The potential for synergism among the pollutants raises concerns about potential adverse health effects, and there are still many uncertainties involved in exposure assessment. In this study, conventional (culture-based) and molecular real-time polymerase chain reaction (RTPCR) methodologies were used to assess fungal air contamination in a waste-sorting plant which focused on the presence of three potential pathogenic/toxigenic fungal species: Aspergillus flavus, A. fumigatus, and Stachybotrys chartarum. In addition, microbial volatile organic compounds (MVOC) were measured by photoionization detection. For all analysis, samplings were performed at five different workstations inside the facilities and also outdoors as a reference. Penicillium sp. were the most common species found at all plant locations. Pathogenic/toxigenic species (A. fumigatus and S. chartarum) were detected at two different workstations by RTPCR but not by culture-based techniques. MVOC concentration indoors ranged between 0 and 8.9 ppm (average 5.3 ± 3.16 ppm). Our results illustrated the advantage of combining both conventional and molecular methodologies in fungal exposure assessment. Together with MVOC analyses in indoor air, data obtained allow for a more precise evaluation of potential health risks associated with bioaerosol exposure. Consequently, with this knowledge, strategies may be developed for effective protection of the workers.

Fungal contamination of poultry litter: a public health problem

Although numerous studies have been conducted on microbial contaminants associated with various stages related to poultry and meat products processing, only a few reported on fungal contamination of poultry litter. The goals of this study were to (1) characterize litter fungal contamination and (2) report the incidence of keratinophilic and toxigenic fungi presence. Seven fresh and 14 aged litter samples were collected from 7 poultry farms. In addition, 27 air samples of 25 litters were also collected through impaction method, and after laboratory processing and incubation of collected samples, quantitative colony-forming units (CFU/m3) and qualitative results were obtained. Twelve different fungal species were detected in fresh litter and Penicillium was the most frequent genus found (59.9%), followed by Alternaria (17.8%), Cladosporium (7.1%), and Aspergillus (5.7%). With respect to aged litter, 19 different fungal species were detected, with Penicillium sp. the most frequently isolated (42.3%), followed by Scopulariopsis sp. (38.3%), Trichosporon sp. (8.8%), and Aspergillus sp. (5.5%). A significant positive correlation was found between litter fungal contamination (CFU/g) and air fungal contamination (CFU/m3). Litter fungal quantification and species identification have important implications in the evaluation of potential adverse health risks to exposed workers and animals. Spreading of poultry litter in agricultural fields is a potential public health concern, since keratinophilic (Scopulariopsis and Fusarium genus) as well as toxigenic fungi (Aspergillus, Fusarium, and Penicillium genus) were isolated.

Supporting leadership development by exposure to role models: two case studies

The idiomatic expression “In Rome be a Roman” can be applied to leadership training and development as well. Leaders who can act as role models inspire other future leaders in their behaviour, attitudes and ways of thinking. Based on two examples of current leaders in the fields of Politics and Public Administration, I support the idea that exposure to role models during their training was decisive for their career paths and current activities as prominent characters in their profession. Issues such as how students should be prepared for community or national leadership as well as cross-cultural engagement are raised here. The hypothesis of transculturalism and cross-cultural commitment as a factor of leadership is presented. Based on current literature on Leadership as well as the presented case studies, I expect to raise a debate focusing on strategies for improving leaders’ training in their cross-cultural awareness.

Development of a biosensor for urea assay based on amidase inhibition, using an ion-selective electrode

A biosensor for urea has been developed based on the observation that urea is a powerful active-site inhibitor of amidase, which catalyzes the hydrolysis of amides such as acetamide to produce ammonia and the corresponding organic acid. Cell-free extract from Pseudomonas aeruginosa was the source of amidase (acylamide hydrolase, EC 3.5.1.4) which was immobilized on a polyethersulfone membrane in the presence of glutaraldehyde; anion-selective electrode for ammonium ions was used for biosensor development. Analysis of variance was used for optimization of the biosensorresponse and showed that 30 mu L of cell-free extract containing 7.47 mg protein mL(-1), 2 mu L of glutaraldehyde (5%, v/v) and 10 mu L of gelatin (15%, w/v) exhibited the highest response. Optimization of other parameters showed that pH 7.2 and 30 min incubation time were optimum for incubation ofmembranes in urea. The biosensor exhibited a linear response in the range of 4.0-10.0 mu M urea, a detection limit of 2.0 mu M for urea, a response timeof 20 s, a sensitivity of 58.245 % per mu M urea and a storage stability of over 4 months. It was successfully used for quantification of urea in samples such as wine and milk; recovery experiments were carried out which revealed an average substrate recovery of 94.9%. The urea analogs hydroxyurea, methylurea and thiourea inhibited amidase activity by about 90%, 10% and 0%, respectively, compared with urea inhibition.

Development of novel ionic liquids based on ampicillin

Novel ionic liquids containing ampicillin as an active pharmaceutical ingredient anion were prepared with good yields by using a new, efficient synthetic procedure based on the neutralization of a moderately basic ammonia solution of ampicillin with different organic cation hydroxides. The relevant physical and thermal properties of these novel ionic liquids based on ampicillin were also evaluated.

Determination of chitin content in fungal cell wall: an alternative flow cytometric method

The conventional methods used to evaluate chitin content in fungi, such as biochemical assessment of glucosamine release after acid hydrolysis or epifluorescence microscopy, are low throughput, laborious, time-consuming, and cannot evaluate a large number of cells. We developed a flow cytometric assay, efficient, and fast, based on Calcofluor White staining to measure chitin content in yeast cells. A staining index was defined, its value was directly related to chitin amount and taking into consideration the different levels of autofluorecence. Twenty-two Candida spp. and four Cryptococcus neoformans clinical isolates with distinct susceptibility profiles to caspofungin were evaluated. Candida albicans clinical isolate SC5314, and isogenic strains with deletions in chitin synthase 3 (chs3Δ/chs3Δ) and genes encoding predicted Glycosyl Phosphatidyl Inositol (GPI)-anchored proteins (pga31Δ/Δ and pga62Δ/Δ), were used as controls. As expected, the wild-type strain displayed a significant higher chitin content (P < 0.001) than chs3Δ/chs3Δ and pga31Δ/Δ especially in the presence of caspofungin. Ca. parapsilosis, Ca. tropicalis, and Ca. albicans showed higher cell wall chitin content. Although no relationship between chitin content and antifungal drug susceptibility phenotype was found, an association was established between the paradoxical growth effect in the presence of high caspofungin concentrations and the chitin content. This novel flow cytometry protocol revealed to be a simple and reliable assay to estimate cell wall chitin content of fungi.

Fungal contamination in swine: a potential occupational health threat

Poor air quality in a pig-confinement building may potentially place farmers at higher health risk than other workers for exposure to airborne pollutants that may reach infectious levels. The aim of this study was to assess worker exposure to fungi in indoor environments in Portuguese swine buildings. Air samples from 7 swine farms were collected at a flow rate of 140 L/min, at 1 m height, onto malt extract agar supplemented with chloramphenicol (MEA). Surfaces samples of the same indoor sites were obtained by swabbing the surfaces. Samples from the floor covering were also collected from four of seven swine farms. All collected samples were incubated at 27°C for 5-7 days. After lab processing and incubation of obtained samples, quantitative colony-forming units (CFU)/m(3), CFU/cm(2), and CFU/g and qualitative results were determined with identification of isolated fungal species. Aspergillus versicolor was the most frequent species found in air (21%), followed by Scopulariopsis brevicaulis (17%) and Penicillium sp. (14%). Aspergillus versicolor was also the most frequent species noted on surfaces (26.6%), followed by Cladosporium sp. (22.4%) and Scopulariopsis brevicaulis (17.5%). Chrysosporium was the most frequently found genera in the new floor covering (38.5%), while Mucor was the most prevalent genera (25.1%) in used floor covering. Our findings corroborate a potential occupational health threat due to fungi exposure and suggest the need for a preventive strategy.

International sustainable development cooperation of physiotherapy course from Escola Superior de Tecnologia da Saúde de Lisboa

Objectives - To share with delegates the international experience in several Program of Physiotherapy course from Escola Superior de Tecnologia da Saúde de Lisboa (ESTeSL).

Effects of fungal contamination on respiratory symptoms of poultry workers

Exposure to certain fungi (molds) can cause human illness by 3 specific mechanisms: generation of a harmful immune response, direct infection by the organism or/and toxic-irritant effects from mold byproducts. Moulds are considered central elements in daily exposure of poultry workers and can be the cause of an increased risk of occupational respiratory diseases, like allergic and non-allergic rhinitis and asthma.

Occupational exposure to toxigenic Aspergillus versicolor in Portuguese swines

Biological factors associated with airbome dust are the most important hazards in pig buildings and include allergenic and/or toxic compounds, as well as infectious agents such as fungi and their metabolites, like mycotoxins. Inhalation of such agents can be a potential occupationai treat. Exposure of workers from swine confinement buildings to respiratory hazards has been reported elsewhere in Europe, Asia and America. Analogous data has not been reported for Portugal and this omission has hindered the development of policies in the area of occupational health and farm safety. Aspergilius versicolor is known as being the major producer of the hepatotoxic and carcinogenic mycotoxin sterigmatocystin. The toxicity of this mycotoxin is manifested primarily in liver and kidney. This study aimed to determine occupational exposure treat due to fungal contamination caused by A. versicolor in seven Portuguese swine.