Wrinkling Labyrinth Patterns on Elastomeric Janus Particles

We describe a novel, low-cost and low-tech method for the fabrication of elastomeric Janus particles with diameters ranging from micrometers to millimeters. This consists of UV-irradiating soft urethane/urea elastomer spheres, which are then extracted in toluene and dried. The spheres are thus composed of a single material: no coating or film deposition steps are required. Furthermore, the whole procedure is carried out at ambient temperature and pressure. Long, labyrinthine corrugations ("wrinkles") appear on the irradiated portions of the particles' surfaces, the spatial periodicity of which can be controlled by varying the sizes of particles. The asymmetric morphology of the resulting Janus particles has been confirmed by scanning electron microscopy, atomic force microscopy, and optical microscopy. We have also established that the spheres behave elastically by performing bouncing tests with dried and swollen spheres. Results can be interpreted by assuming that each sphere consists of a thin, stiff surface layer ("skin") lying atop a thicker, softer substrate ("bulk"). The skin's higher stiffness is hypothesized to result from the more extensive cross-linking of the polymer chains located near the surface by the UV radiation. Textures then arise from competition between the effects of bending the skin and compressing the bulk, as the solvent evaporates and the sphere shrinks.

Exposure to dust in poultry: the importance of task differences for detailed exposure assessment

Dust is a complex mixture of particles of organic and inorganic origin and different gases absorbed in aerosol droplets. In a poultry unit include dried faecal matter and urine, skin flakes, ammonia, carbon dioxide, pollens, feed and litter particles, feathers, grain mites, fungi spores, bacteria, viruses and their constituents. Dust particles vary in size and differentiation between particle size fractions is important in health studies in order to quantify penetration within the respiratory system. A descriptive study was developed in order to assess exposure to particles in a poultry unit during different operations, namely routine examination and floor turn over. Direct-reading equipment was used (Lighthouse, model 3016 IAQ). Particle measurement was performed in 5 different sizes (PM0.5; PM1.0; PM2.5; PM5.0; PM10). The chemical composition of poultry litter was also determined by neutron activation analysis. Normally, the litter of poultry pavilions is turned over weekly and it was during this operation that the higher exposure of particles was observed. In all the tasks considered PM5.0 and PM10.0 were the sizes with higher concentrations values. PM10 is what turns out to have higher values and PM0.5 the lowest values. The chemical element with the highest concentration was Mg (5.7E6 mg.kg-1), followed by K (1.5E4 mg.kg-1), Ca (4.8E3 mg.kg-1), Na (1.7E3 mg.kg-1), Fe (2.1E2 mg.kg-1) and Zn (4.2E1 mg.kg-1). This high presence of particles in the respirable range (<5–7μm) means that poultry dust particles can penetrate into the gas exchange region of the lung. Larger particles (PM10) present a range of concentrations from 5.3E5 and 3.0E6 mg/m3.

A novel flow cytometric protocol for assessment of yeast cell adhesion

Microbial adhesion is a field of recognized relevance and, as such, an impressive array of tools has been developed to understand its molecular mechanisms and ultimately for its quantification. Some of the major limitations found within these methodologies concern the incubation time, the small number of cells analyzed, and the operator's subjectivity. To overcome these aspects, we have developed a quantitative method to measure yeast cells' adhesion through flow cytometry. In this methodology, a suspension of yeast cells is mixed with green fluorescent polystyrene microspheres (uncoated or coated with host proteins). Within 2 h, an adhesion profile is obtained based on two parameters: percentage and cells-microsphere population's distribution pattern. This flow cytometry protocol represents a useful tool to quantify yeast adhesion to different substrata in a large scale, providing manifold data in a speedy and informative manner.

Optimization of immunocytochemistry in cytology: comparison of two protocols for fixation and preservation on cytospin and smear preparations

Objective: A new protocol for fixation and slide preservation was evaluated in order to improve the quality of immunocytochemical reactions on cytology slides. Methods: The quality of immunoreactions was evaluated retrospectively on 186 cytology slides (130 direct smears, 56 cytospins) prepared from different cytology samples. Ninety-three of the slides were air dried, stored at -20 °C and fixed in acetone for 10 minutes (Protocol 1), whereas the other 93 were immediately fixed in methanol at -20 °C for at least 30 minutes, subsequently protected with polyethylene glycol (PEG) and stored at room temperature (Protocol 2). Immunocytochemical staining, with eight primary antibodies, was performed on a Ventana BenchMark Ultra instrument using an UltraView Universal DAB Detection Kit. The following parameters were evaluated for each immunoreaction: morphology preservation, intensity of specific staining, background and counterstain. The slides were blinded and independently scored by four observers with marks from 0 to 20. Results: The quality of immunoreactions was better on methanol-fixed slides protected with PEG than on air-dried slides stored in the freezer: X¯ = 14.44 ± 3.58 versus X¯ = 11.02 ± 3.86, respectively (P < 0.001). Conclusion: Immediate fixation of cytology slides in cold methanol with subsequent application of PEG is an easy and straightforward procedure that improves the quality of immunocytochemical reactions and allows the storage of the slides at room temperature.

Gamma radiation effects on microbial inactivation of two medicinal plants

The consumption of natural products has become a public health problem, since these medicinal teas are prepared using natural plants without an effective hygienic and sanitary control. The aim of this study was to assess the effects of gamma radiation, on the microbial burden of two medicinal plants: Melissa officinalis and Lippia citriodora. Dried samples of the two plants were irradiated at a Co-60 experimental equipment. The applied gamma radiation doses were 1, 3, and 5 kGy at a dose rate of 1.34 kGy/h. Non-irradiated samples followed all the experiments. Bacterial and fungal counts were assessed before and after irradiation by membrane filtration method. Challenging tests with Escherichia coli were performed in order to evaluate the disinfection efficiency of gamma radiation treatment. Characterization of M. officinalis and L. citriadora microbiota indicated an average bioburden value of 102CFU/g. The inactivation studies of the bacterial mesophilic population of both dried plants pointed out to a one log reduction of microbial load after irradiation at 5 kGy. Regarding the fungal population, the initial load of 30 CFU/g was only reduced by 0.5 log by an irradiation dose of 5 kGy. The dynamics with radiation doses of plants microbial population’s phenotypes indicated the prevalence of gram-positive rods for M. officinalis before and after irradiation, and the increase of the frequency of gram-negative rods with irradiation for L. citriadora. Among fungal population of both plants, Mucor, Neoscytalidium, Aspergillus and Alternaria were the most isolated genera. The results obtained in the challenging tests with E. coli on plants pointed out to an inactivation efficiency of 99.5% and 99.9% to a dose of 2 kGy, for M.officinalis and L. citriadora, respectively. The gamma radiation treatment can be a significant tool for the microbial control in medicinal plants.

Sistemas de arrefecimento de instalações frigoríficas utilizando três agentes frigoríficos

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Mecânica

Impact of a training course on the quality of malaria diagnosis by microscopy in Angola

Background: In Angola, malaria is an endemic disease having a major impact on the economy. The WHO recommends testing for all suspected malaria cases, to avoid the presumptive treatment of this disease. In malaria endemic regions laboratory technicians must be very comfortable with microscopy, the golden standard for malaria diagnosis, to avoid the incorrect diagnosis. The improper use of medication promotes drug resistance and undesirable side effects. The present study aims to assess the impact of a three-day refresher course on the knowledge of technicians, quality of blood smears preparation and accuracy of microscopy malaria diagnosis, using qPCR as reference method. Methods: This study was implemented in laboratories from three hospitals in different provinces of Angola: Bengo, Benguela and Luanda. In each laboratory samples were collected before and after the training course (slide with thin and thick blood smears, a dried blood spot and a form). The impact of the intervention was evaluated through a written test, the quality of slide preparation and the performance of microscopy. Results: It was found a significant increase on the written test median score, from 52.5% to 65.0%. A total of 973 slides were analysed to evaluate the quality of thick and thin blood smears. Considering all laboratories there was a significant increase in quality of thick and thin blood smears. To determine the performance of microscopy using qPCR as the reference method we used 1,028 samples. Benguela presented the highest values for specificity, 92.9% and 98.8% pre and post-course, respectively and for sensitivity the best pre-course was Benguela (75.9%) and post-course Luanda (75.0%). However, no significant increase in sensitivity and specificity after the training course was registered in any laboratory analysed. Discussion: The findings of this study support the need of continuous refresher training for microscopists and other laboratory staff. The laboratories should have a quality control programme to supervise the diagnosis and also to assess the periodicity of new training. However, other variables needed to be considered to have a correct malaria diagnosis, such as adequate equipment and reagents for staining and visualization, good working conditions, motivated and qualified personnel.

Chemical composition and antioxidante activity of Tunisian Origanum glandulosum Desf. essential oil and volatile oil obtain by supercritical CO2 extraction

Dried flowers and leaves of Origanum glandulosum Desf. were submitted to hydrodistillation (HD) and supercritical fluid extraction with CO2 (SFE). The essential oils isolated by HD and volatile oils obtained by SFE were analysed by GC and GC/MS. Total phenolics content and antioxidant effectiveness were performed. The main components of the essential oils from Bargou and Nefza were: p-cymene (40.4% and 39%), thymol (38.7% and 34.4%) and γ- terpinene (12.3% and 19.2%), respectively. The major components obtain by SFE in the volatile oil, from Bargou and Nefza, were: p-cymene (32.3% and 36.2%), thymol (41% and 40%) and γ-terpinene (20.3% and 13.3%). Total phenolic content, expressed in gallic acid equivalent (GAE) g kg-1 dry weight, varied from 12 to 27 g kg-1 dw, and the ability to scavenge the DPPH radicals, expressed by IC50 ranged from 44 to143 mg L-1.

Hierarchical wrinkling on elastometric Janus spheres

Hierarchical wrinkling on elastomeric Janus spheres is permanently imprinted by swelling, for different lengths of time, followed by drying the particles in an appropriate solvent. First-order buckling with a spatial periodicity (lambda(11)) of the order of a few microns and hierarchical structures comprising of 2nd order buckling with a spatial periodicity (lambda(12)) of the order of hundreds of nanometers have been obtained. The 2nd order buckling features result from a Grinfeld surface instability due to the diffusion of the solvent and the presence of sol molecules.

Electrorheological characterization of dispersions in silicone oil of encapsulated liquid crystal 4-n-penthyl-4 '-cyanobiphenyl in polyvinyl alcohol and in silica

The electrorheological (ER) effect is known as the change in the apparent viscosity upon the application of an external electric field perpendicular to the flow direction. In this work we present the electrorheological behaviour of suspensions in silicone oil of two different dispersed phases: foams of liquid crystal 4-n-penthyl-4'-cyanobiphenyl (5CB) encapsulated in polyvinyl alcohol (PVA) and nano/microspheres of 5CB encapsulated in silica. We will present the viscosity curves under the application of an electric field ranging between 0 and 3 kV mm(-1). The ER effect was observed for the suspensions of 5CB/PVA but not in the case of 5CB/silica. For the case of the suspensions of 5CB/PVA, the effect of the viscosity of the continuum phase and the concentration of the dispersed phase was analysed, showing that the enhancement of the viscosity of the suspension increases with the concentration, as expected, however the continuum phase viscosity has no significant effect, at least in the investigated viscosity range.