Human vaccinia-like virus outbreaks in São Paulo and Goiás States, Brazil: virus detection, isolation and identification

Since October 2001, the Adolfo Lutz Institute has been receiving vesicular fluids and scab specimens of patients from Paraíba Valley region in the São Paulo and Minas Gerais States and from São Patricio Valley, in the Goiás State. Epidemiological data suggested that the outbreaks were caused by Cowpox virus or Vaccinia virus. Most of the patients are dairy milkers that had vesiculo-pustular lesions on the hands, arms, forearms, and some of them, on the face. Virus particles with orthopoxvirus morphology were detected by direct electron microscopy (DEM) in samples of 49 (66.21%) patients of a total of 74 analyzed. Viruses were isolated in Vero cell culture and on chorioallantoic membrane (CAM) of embryonated chicken eggs. Among 21 samples submitted to PCR using primers for hemagglutinin (HA) gene, 19 were positive. Restriction digestion with TaqI resulted in four characteristic Vaccinia virus fragments. HA nucleotide sequences showed 99.9% similarity with Cantagalo virus, described as a strain of Vaccinia virus. The only difference observed was the substitution of one nucleotide in the position 616 leading to change in one amino acid of the protein in the position 206. The phylogenetic analysis showed that the isolates clustered together with Cantagalo virus, other Vaccinia strains and Rabbitpox virus.

Heparinized nanohydroxyapatite/collagen granules for controlled release of vancomycin

The purpose of this study was to develop a bone substitute material capable of preventing or treating osteomyelitis through a sustainable release of vancomycin and simultaneously inducing bone regeneration. Porous heparinized nanohydroxyapatite (nanoHA)/collagen granules were characterized using scanning electron microscopy, micro-computed tomography and attenuated total reflectance Fourier transform infrared spectroscopy. After vancomycin adsorption onto the granules, its releasing profile was studied by UV molecular absorption spectroscopy. The heparinized granules presented a more sustainable release over time, in comparison with nonheparinized nanoHA and nanoHA/collagen granules. Vancomycin was released for 360 h and proved to be bioactive until 216 h. Staphylococcus aureus adhesion was higher on granules containing collagen, guiding the bacteria to the material with antibiotic, improving their eradication. Moreover, cytotoxicity of the released vancomycin was assessed using osteoblast cultures, and after 14 days of culture in the presence of vancomycin, cells were able to remain viable, increasing their metabolic activity and colonizing the granules, as observed by scanning electron microscopy and confocal laser scanning microscopy. These findings suggest that heparinized nanoHA/collagen granules are a promising material to improve the treatment of osteomyelitis, as they are capable of releasing vancomycin, eliminating the bacteria, and presented morphological and chemical characteristics to induce bone regeneration.

Molecularly imprinted sensor for voltammetric detection of norfloxacin

In this work, a norfloxacin selective modified glassy carbon electrode (GCE) based on a molecularly imprinted polymer (MIP) as electrochemical sensor was developed. A suspension of multi-walled carbon nanotubes (MWCNTs) was deposited on the electrode surface. Subsequently, a molecularly imprinted film was prepared by electropolymerization, via cyclic voltammetry of pyrrole (PPy) in the presence of norfloxacin (NFX) as the template molecule. A control electrode (NIP) was also prepared. Scanning electron microscopy (SEM) and cyclic voltammetry in a ferrocyanide solution were performed for morphological and electrochemical characterisation, respectively. Several experimental parameters were studied and optimised. For quantification purposes the MIP/MWCNT/GCE was immersed in NFX solutions for 10 min, and the detection was performed in voltammetric cell by square wave voltammetry. The proposed sensor presented a linear behaviour, between peak current intensity and logarithmic concentration of NFX between 1 × 10−7 and 8 × 10−6 M. The obtained results presented good precision, with a repeatability of 4.3% and reproducibility of 9% and the detection limit was 4.6 × 10−8 M (S/N = 3). The developed sensor displayed good selectivity and operational lifetime, is simple to fabricate and easy to operate and was successfully applied to the analysis of NFX in urine samples.

Nocardiosis: an overview and additional report of 28 cases in cattle and dogs

Phenotypic characteristics, antimicrobial susceptibility profile, and clinical-epidemiological features of 28 Nocardia strains isolated from 19 cases of bovine mastitis, eight cutaneous-subcutaneous lesions and one case of pneumonia in dogs were evaluated. Microbiological, biochemical, cytological and scanning electron microscopy methods were used in diagnosis. Nocardia asteroides type IV, Nocardia otitidiscaviarum,Nocardia nova (type III) and Nocardia farcinica (type V) were isolated from bovine milk, bronchial lavage and/or cutaneous-subcutaneous abscesses in dogs. Nocardial bovine mastitis was diagnosed predominantly in clinical cases, in dairy herds with poor environmental hygienic conditions between milking and inappropriate intramammary therapy. Canine nocardiosis was observed commonly in animals co-infected with distemper virus. Sulphamethoxazole-trimethoprim (92.8%), amikacin (92.8%) and ceftiofur (92.8%) were the most effective drugs in 28 isolates. Multiple drug resistance to three or more and five or more antimicrobials was observed in ten (35.7%) and three (10.7%) strains, respectively, predominantly with use of cloxaxillin, cefoperazone and ampicillin. The species (type) classification, clinical-epidemiological characteristics, diagnosis, multiple-drug resistance and public health considerations in Nocardia strains isolated from cattle and dogs in Brazil are discussed, with special reference to report of bovine mastitis by N. otitidiscaviarum by first time in Brazil and the similarity between Nocardia species isolated from human and animal origin.

Structural adhesives modified with thermally expandable particles

In this study, the behaviour of two structural adhesives modified with thermally expandable particles (TEPs) was investigated as a preliminary study for further investigations on the potential of TEPs in adhesive joints. Tensile bulk tests were performed to get the tensile properties of the adhesives and TEPs-modified adhesives. In order to determine the expansion temperature of the particles while encapsulated in these particular adhesive systems, the variation of the volume of adhesive samples modified with different TEPs concentration as a function of temperature was measured. Further, the possibility of any chemical interactions between TEPs and adhesives matrix in the TEPs-modified specimens was verified by a Fourier transform infrared spectroscopy analysis. Finally, the fracture surfaces of the unmodified and TEPs-modified specimens, as well as the dispersion and the morphology of the particles, were examined by a scanning electron microscopy analysis. It was found that the stiffness of the TEPs-modified adhesives is not affected by incorporation of TEPs in the adhesives matrix, while the tensile yield strength decreased by increasing the wt% TEPs content. In applications of such particular materials (TEPs-modified adhesives), the temperature should be controlled to stay between 90°C and 120°C in order to obtain the highest expansion ratio. At a lower temperature, not all the particles will expand, and above, the TEPs will deteriorate and as a result the TEPs-modified adhesives will deteriorate.

An outbreak of scalp white piedra in a Brazilian children day care

White piedra is a superficial mycosis caused by Trichosporon spp. that affects the hair shaft of any part of the body. It is presented an outbreak of scalp white piedra seen in 5.8% of the children frequenting a day care in Northeastern of São Paulo State, Brazil. Mycological exam and culture identified T. cutaneum in all five cases, and scanning electron microscopy of nodules around hair shaft infected by Trichosporon spp. is demonstrated comparing them with those of black piedra and with nits of Pediculous capitis.

Clinical and Pathological Findings in Women with Fabry Disease

Introduction. Fabry disease is a rare metabolic disorder caused by the genetic deficiency of the lysosomal hydrolase alpha-galactosidase A, located on chromosome X. Females with the defective gene are more than carriers and can develop a wide range of symptoms. Nevertheless, disease symptoms generally occur later and are less severe in women than in men. The enzyme deficiency manifests as a glycosphingolipidosis with progressive accumulation of glycosphingolipids and deposit of inclusion bodies in lysosomes giving a myelinlike appearance. Patients and Methods. Records of renal biopsies performed on adults from 1st January 2008 to 31st August 2011, were retrospectively examined at the Renal Pathology Laboratory. We retrieved biopsies diagnosed with Fabry disease and reviewed clinical and laboratory data and pathology findings. Results. Four female patients with a mean age of 49.3±4.5 (44-55) years were identified. The mean proteinuria was 0.75±0.3 g/24h (0.4-1.2) and estimated glomerular filtration rate (CKD EPI equation) was 71±15.7 ml/min/1.73m2 (48-83). Three patients experienced extra-renal organ involvement (cerebrovascular, cardiac, dermatologic, ophthalmologic and thyroid) with distinct severity degrees. Leukocyte α-GAL A activity was below normal range in the four cases but plasma and urinary enzymatic activity was normal. Light microscopy showed predominant vacuolisation of the podocyte cytoplasm and darkly staining granular inclusions on paraffin and plastic-embedded semi-thin sections. Electron microscopy showed in three patients the characteristic myelin-like inclusions in the podocyte cytoplasm and also focal podocyte foot process effacement. In one case the inclusions were also present in parietal glomerular cells, endothelial cells of peritubular capillary and arterioles. Conclusion. Clinical signs and symptoms are varied and can be severe among heterozygous females with Fabry disease. Intracellular accumulation of glycosphingolipids is a characteristic histologic finding of Fabry nephropathy. Since this disease is a potentially treatable condition, its early identification is imperative. We should consider it in the differential diagnosis of any patient presenting with proteinuria and/or chronic kidney disease, especially if there is a family history of kidney disease.

Effect of selenization conditions on the growth and properties of Cu2ZnSn(S,Se)4 thin films

The opto-electronic properties of copper zinc tin sulfide can be tuned to achieve better cell efficiencies by controlled incorporation of selenium. In this paper we report the growth of Cu2ZnSn(S,Se)4 (CZTSSe) using a hybrid process involving the sequential evaporation of Zn and sputtering of the sulfide precursors of Cu and Sn, followed by a selenization step. Two approaches for selenization were followed, one using a tubular furnace and the other using a rapid thermal processor. The effects of annealing conditions on the morphological and structural properties of the films were investigated. Scanning electron microscopy and energy dispersive spectroscopy were employed to investigate the morphology and composition of the films. Structural analyses were done using X-ray diffraction (XRD) and Raman spectroscopy. Structural analyses revealed the formation of CZTSSe. This study shows that regardless of the selenization method a temperature above 450 °C is required for conversion of precursors to a compact CZTSSe layer. XRD and Raman analysis suggests that the films selenized in the tubular furnace are selenium rich whereas the samples selenized in the rapid thermal processor have higher sulfur content.

A comparison between thin film solar cells made from co-evaporated CuIn1-xGaxSe2using a one-stage process versus a three-stage process

Until this day, the most efficient Cu(In,Ga)Se2 thin film solar cells have been prepared using a rather complex growth process often referred to as three-stage or multistage. This family of processes is mainly characterized by a first step deposited with only In, Ga and Se flux to form a first layer. Cu is added in a second step until the film becomes slightly Cu-rich, where-after the film is converted to its final Cu-poor composition by a third stage, again with no or very little addition of Cu. In this paper, a comparison between solar cells prepared with the three-stage process and a one-stage/in-line process with the same composition, thickness, and solar cell stack is made. The one-stage process is easier to be used in an industrial scale and do not have Cu-rich transitions. The samples were analyzed using glow discharge optical emission spectroscopy, scanning electron microscopy, X-ray diffraction, current–voltage-temperature, capacitance-voltage, external quantum efficiency, transmission/reflection, and photoluminescence. It was concluded that in spite of differences in the texturing, morphology and Ga gradient, the electrical performance of the two types of samples is quite similar as demonstrated by the similar J–V behavior, quantum spectral response, and the estimated recombination losses.

Archaeometallurgical study of pre and protohistoric production remains and artefacts from Southern Portugal

Dissertação para obtenção do Grau de Doutor em Conservação e Restauro, especialidade Ciências da Conservação

Brachial Plexus Morphology and Vascular Supply in the Wistar Rat

Introduction: The rat is probably the animal species most widely used in experimental studies on nerve repair. The aim of this work was to contribute to a better understanding of the morphology and blood supply of the rat brachial plexus. Material and Methods: Thirty adult rats were studied regarding brachial plexus morphology and blood supply. Intravascular injection and dissection under an operating microscope, as well as light microscopy and scanning electron microscopy techniques were used to define the microanatomy of the rat brachial plexus and its vessels. Results: The rat brachial plexus was slightly different from the human brachial plexus. The arterial and venous supply to the brachial plexus plexus was derived directly or indirectly from neighboring vessels. These vessels formed dense and interconnected plexuses in the epineurium, perineurium, and endoneurium. Several brachial plexus components were accompanied for a relatively long portion of their length by large and constant blood vessels that supplied their epineural plexus, making it possible to raise these nerves as flaps. Discussion: The blood supply to the rat brachial plexus is not very different from that reported in humans, making the rat a useful animal model for the experimental study of peripheral nerve pathophysiology and treatment. Conclusion: Our results support the homology between the rat and the human brachial plexus in terms of morphology and blood supply. This work suggests that several components of the rat brachial plexus can be used as nerve flaps, including predominantly motor, sensory or mixed nerve fibers. This information may facilitate new experimental procedures in this animal model.

Materiais e dispositivos sensores para a Norfloxacina

O trabalho consistiu no desenvolvimento e caracterização de sensores potenciométricos com base em polímeros de impressão molecular para a determinação de um antibiótico, a norfloxacina, em aquacultura. A simplicidade, o baixo custo e a interação rápida e reversível dos sensores potenciométricos com os analitos fizeram com que este fosse o tipo de sensor escolhido. O material sensor foi obtido por tecnologia de impressão molecular, baseada em polimerização em bulk, em que a NOR foi a molécula molde e foram utilizados como monómeros para autoconstrução dos sensores o pirrol, isoladamente, ou em conjunto com partículas de sílica gel funcionalizadas com 3-aminopropil. Também foi obtido material sensor, para controlo, em que a molécula molde NOR não estava presente (NIP). As características dos materiais sensores foram sujeitas a análise de microscopia eletrónica SEM e análise por espectrómetro de infravermelhos com transformada de Fourier. Os materiais sensores foram incluídos em membranas poliméricas, que seriam incorporadas em elétrodos. A avaliação do desempenho dos elétrodos foi feita através de curvas de calibração em diferentes meios (PBS, MES e HEPES). Também foi efetuada com sucesso a análise da sensibilidade dos elétrodos em água dopada. As diversas avaliações e análises efetuadas levaram a concluir que o MIP de pirrol com aditivo aniónico, foi o material sensor testado que permitiu obter melhores propriedades de resposta.

ULTRASTRUCTURAL CHANGES IN Schistosoma mansoni MALE WORMS AFTER in vitro INCUBATION WITH THE ESSENTIAL OIL OF Mentha x villosa Huds

Introduction: The essential oil Mentha x villosa (MVEO) has a wide range of actions, including antibacterial, antifungal, antiprotozoal and schistosomicidal actions. The present study aimed to investigate the ultrastructural changes of MVEO on the tegument of adult Schistosoma mansoni. Materials and Methods: Different concentrations of MVEO were tested on S. mansoni adult worms in vitro. Ultrastructural changes on the tegument of these adult worms were evaluated using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Results: The MVEO caused the death of all worms at 500 μg mL-1 after 24 h. After 24h of 500 μg mL-1 MVEO treatment, bubble lesions were observed over the entire body of worms and they presented loss of tubercles in some regions of the ventral portion. In the evaluation by TEM, S. mansoni adult worms treated with MVEO, 500 μg mL-1, presented changes in the tegument and vacuoles in the syncytial matrix region. Glycogen granules close to the muscle fibers were visible. Conclusion: The ability of MVEO to cause extensive ultrastructural damage to S. mansoni adult worms correlates with its schistosomicidal effects and confirms earlier findings with S. mansoni.

BIOFILM FORMATION OF Vibrio cholerae ON STAINLESS STEEL USED IN FOOD PROCESSING

Vibrio cholerae represents a significant threat to human health in developing countries. This pathogen forms biofilms which favors its attachment to surfaces and its survival and transmission by water or food. This work evaluated the in vitro biofilm formation of V. cholerae isolated from clinical and environmental sources on stainless steel of the type used in food processing by using the environmental scanning electron microscopy (ESEM). Results showed no cell adhesion at 4 h and scarce surface colonization at 24 h. Biofilms from the environmental strain were observed at 48 h with high cellular aggregations embedded in Vibrio exopolysaccharide (VPS), while less confluence and VPS production with microcolonies of elongated cells were observed in biofilms produced by the clinical strain. At 96 h the biofilms of the environmental strain were released from the surface leaving coccoid cells and residual structures, whereas biofilms of the clinical strain formed highly organized structures such as channels, mushroom-like and pillars. This is the first study that has shown the in vitro ability of V. cholerae to colonize and form biofilms on stainless steel used in food processing.

Application of ionic liquids and enzymes for the removal of proteinaceous layers from polychrome of works of art and evaluation of the cleaning effectiveness

Dissertação Apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Ciências da Conservação, especialização em Pintura