Pteridine reductase (PTR1): initial structure-activity relationships studies of potential leishmanicidal arylindole derivatives compounds.

Leishmaniasis is a public health concern, especially in Brazil and India. The drugs available for therapy are old, cause toxicity and have reports of resistance. Therefore, this paper aimed to carry out initial structure-activity relationships (applying molecular docking and dynamic simulations) of arylindole scaffolds against the pteridine reductase (PTR1), which is essential target for the survival of the parasite. Thus, we used a series of 43 arylindole derivatives as a privileged skeleton, which have been evaluated previously for different biological actions. Compound 7 stood out among its analogues presenting the best results of average number of interactions with binding site (2.00) and catalytic triad (1.00). Additionally, the same compound presented the best binding free energy (-32.33 kcal/mol) in dynamic simulations. Furthermore, with computational studies, it was possible to comprehend and discuss the influences of the substituent sizes, positions of substitutions in the aromatic ring and electronic influences. Therefore, this study can be a starting point for the structural improvements needed to obtain a good leishmanicidal drug.

New 1,3,4-oxadiazole compound with effective antibacterial and antibiofilm activity against Staphylococcus aureus.

Staphylococcus aureus is one of the main aetiological agents causing food-borne diseases. Some strains produce enterotoxins responsible for food poisoning. In addition, they can form biofilms on several surfaces such as plastics, glass and stainless steel making it difficult to eliminate them. The present study evaluated, for the first time, the antibacterial and antibiofilm activities of the synthetic compound LMM6 against S. aureus. The minimum inhibitory concentration was 0·97, 1·95 and 1·95 µg ml-1 against S. aureus ATCC 25923, S. aureus 629/94 and S. aureus FRI S-6, respectively. The time-kill curves showed that 96 h treatment with LMM6 reduced approximately 4 log CFU per ml at all tested concentrations. Furthermore, LMM6 reduced S. aureus preformed biofilm by approximately 1 log CFU per cm2 . During biofilm formation, a reduction of approximately 4 log CFU per cm2 was observed. LMM6 also reduced biofilm biomass during (~60%) and after biofilm formation (~25 to 45%), as shown by the crystal violet assay. Based on these results, we conclude that LMM6 exhibits antibacterial and antibiofilm activity and may be an innovative synthetic molecule for controlling S. aureus.

Body, metabolic and renal changes following cross-sex estrogen/progestogen therapy in a rodent model simulating its use by transwomen.

PURPOSE: The use of sex steroids by trans people has been of paramount importance regarding body changes during gender transition. The objective of this study was to assess the effects of an injectable steroid combination frequently used by transwomen, namely estradiol enanthate with dihydroxyprogesterone acetophenide (E2EN/DHPA), on blood pressure and metabolic outcomes using an animal model. METHODS: Two-month-old male Wistar rats were orchiectomized or sham-operated and divided into groups: (1) Sham treated with sesame oil vehicle (SG), (2) sham treated with E2EN/DHPA (SP), (3) orchiectomized rats treated with vehicle (OG), and (4) orchiectomized rats treated with E2EN/DHPA (OP), with all groups treated every 10 days for 5 months. We evaluated systolic blood pressure (SBP), body weight (BW), abdominal circumference, nasoanal length (NAL), food and water intake (FI, WI), lipid profile (triglycerides, LDL, and HDL), serum C-reactive protein (CRP), plasma concentrations of urea (URpl) and creatinine (CRpl), 24 h urinary volume (V24 h), sodium and potassium excretion (UNa+, UK+), and proteinuria. RESULTS: E2EN/DHPA administration reduced BW (SP 324.5 ± 31.1; OP 291.7 ± 41.3 g) and NAL (SP 24.5 ± 0.4; OP 24.6 ± 1.0 cm), without changing blood pressure, but increased URpl concentration (SP 55.0 ± 4.8; OP 42.5 ± 8.8 mg/dL) and CRpl (SP 0.47 ± 0.05; OP 0.46 ± 0.04 mg/dL), sodium (SP 3.1 ± 0.8; OP 3.3 ± 0.4 µEq/min/kg), potassium (SP 0.91 ± 0.22; OP 0.94 ± 0.22 µEq/min/kg) excretions and urinary volume (SP 15.5 ± 2.1; OP 16.4 ± 2.9 mL/24 h). CONCLUSION: Cross-sex hormone therapy with E2EN/DHPA significantly modified body characteristics in male rats, producing a feminizing change without altering blood pressure or generating harmful metabolic parameters, but larger translational studies are still needed.

Polysaccharide-based layer-by-layer nanoarchitectonics with sulfated chitosan for tuning anti-thrombogenic properties.

The development of blood-interacting surfaces is critical to fabricate biomaterials for medical use, such as prostheses, implants, biosensors, and membranes. For instance, thrombosis is one of the leading clinical problems when polymer-based materials interact with blood. To overcome this limitation is necessary to develop strategies that limit platelets adhesion and activation. In this work, hyaluronan (HA)/chitosan (Chi) based-films, recently reported in the literature as platforms for tumor cell capture, were developed and, subsequently, functionalized with sulfated chitosan (ChiS) using a layer-by-layer technique. ChiS, when compared to native Chi, presents the unique abilities to confer anti-thrombogenic properties, to reduce protein adsorption, and also to limit calcification. Film physicochemical characterization was carried out using FTIR and XPS for chemical composition assessment, AFM for the surface morphology, and contact angle for hydrophilicity evaluation. The deposition of ChiS monolayer promoted a decrease in both roughness and hydrophilicity of the HA/Chi films. In addition, the appearance of sulfur in the chemical composition of ChiS-functionalized films confirmed the film modification. Biological assay indicated that the incorporation of sulfated groups limited platelet adhesion, mainly because a significant reduction of platelets adhesion to ChiS-functionalized films was observed compared to HA/Chi films. On balance, this work provides a new insight for the development of novel antithrombogenic biomaterials, opening up new possibilities for devising blood-interaction surfaces.

Effects of branched-chain amino acids on glucose uptake and lactose synthesis rates in bovine mammary epithelial cells and lactating mammary tissue slices.

Even though supplementations of essential AA (EAA) are often related to increased lactose yields in dairy cows, underlying mechanisms connecting EAA availability to the mammary glands and lactose synthesis are poorly understood. The objective of this study was to examine the effects of branched-chain AA (BCAA) including Leu, Ile, and Val on (1) glucose transporter (GLUT1) abundance and glucose uptake, (2) the abundance of proteins regulating lactose synthesis pathway, and (3) fractional synthesis rates of lactose (FSR) using bovine mammary epithelial cells (BMEC) and mammary tissues slices (MTS). The BMEC (n = 4) were allocated randomly to regular Dulbecco's Modified Eagle Medium with Ham's F12 (DMEM/F12) medium (+EAA) or +EAA deficient (by 90%) in all EAA (-EAA), all BCAA (-BCAA), only Leu (-Leu), only Ile (-Ile) or only Val (-Val). Western immunoblotting analyses, depletion of glucose in media, and a proteomic analysis were performed to determine the abundance of GLUT1 in the cell membrane, net glucose uptake, and the abundance of enzymes involved in lactose synthesis pathway in BMEC, respectively. The MTS (n = 6) were allocated randomly to DMEM/F12 medium having all EAA and 13C-glucose at concentrations similar to plasma concentrations of cows (+EAAp), and +EAAp deprived of all BCAA (-BCAAp) or only Leu (-Leup) for 3 h. The 13C enrichments of free glucose pool in MTS (EGlu-free) and the enrichments of glucose incorporated into lactose in MTS and media [ELactose-bound (T&M)] were determined and used in calculating FSR. In BMEC, -BCAA increased the fraction of total GLUT1 translocated to the cell membrane and the fraction that was potentially glycosylated compared with +EAA. Among individual BCAA, only -Leu was associated with a 63% increase in GLUT1 translocated to the cell membrane and a 40% increase in glucose uptake of BMEC. The -BCAA tended to be related to a 75% increase in the abundance of hexokinase in BMEC. Deprivation of Leu tended to increase glucose uptake of MTS but did not affect EGlu-free, ELactose-bound (T&M), or FSR relative to +EAAp. On the other hand, -BCAAp did not affect glucose uptake of MTS but was related to lower ELactose-bound (T&M), or FSR relative to +EAAp. Considering together, decreasing Leu supply to mammary tissues enhances GLUT1 and thus glucose uptake, which, however, does not affect lactose synthesis rates. Moreover, the deficiency of other BCAA, Ile, and Val alone or together with the deficiency of Leu seemed to decrease lactose synthesis rates without affecting glucose uptake. The data also emphasize the importance of addressing the effect of the supply of other nutrients to the mammary glands than the precursor supply in describing the synthesis of a milk component.

The Upper Juruá Extractive Reserve in the Brazilian Amazon: past and present / A Reserva Extrativista do Alto Juruá na Amazônia Brasileira: passado e presente

Amazonian livelihoods are largely dependent on rivers, with local protein consumption mainly relying on several species of fish. The UJER (Upper Juruá Extractive Reserve - Reserva Extrativista do Alto Juruá) is located in the state of Acre, bordering Peru and several indigenous areas. Here we summarize the data we collected in 1993/1994 on the population living along the banks of the Juruá, Tejo, Bagé, Igarapé São João and Breu rivers on crop cultivation, animal husbandry, and use of game and fish resources. We interviewed 133 individuals (94 on the Juruá and Tejo, 16 on Bagé, 16 on Igarapé São João and 7 on Breu rivers). Our results include a comprehensive description on local livelihoods, including the most important fish species for local subsistence considering gender and seasonality, the main husbandry and game species, and the items cultivated on the local agriculture. Whenever more recent information was available in the literature, we compared changes in livelihoods over time in the same region and also with the recent patterns observed in the Lower and in the Middle Juruá River. We hope to provide useful information to understand temporal changes in local livelihoods, which can help adapt and shape the ecological management in the region.

Comunidades da Amazônia são amplamente dependentes dos rios, com o consumo local de alimentos baseado em várias espécies de peixes. A UJER (Reserva Extrativista do Alto Juruá - Reserva Extrativista do Alto Juruá) está localizada no estado do Acre, na fronteira com o Peru e com várias áreas indígenas. Aqui resumimos os dados que coletamos em 1993/1994 da população que vive às margens dos rios Juruá, Tejo, Bagé, Igarapé São João e Breu sobre cultivo, criação de animais e uso de recursos de caça e peixes. Foram entrevistados 133 indivíduos (94 nos rios Juruá e Tejo, 16 em Bagé, 16 em Igarapé São João e 7 no rio Breu). Nossos resultados incluem uma descrição abrangente dos meios de subsistência locais, incluindo as espécies de peixes mais importantes para a subsistência local, considerando gênero e sazonalidade, bem como as principais espécies de criação, de caça e os itens cultivados na agricultura local., Quando foi possível obter informações da literatura, comparamos as mudanças nos meios de subsistência ao longo do tempo na mesma região e também com os padrões recentes observados no Baixo e no Médio Rio Juruá. Esperamos fornecer informações úteis para o entendimento das mudanças temporais nos meios de subsistência locais, o que pode ser útil ao manejo ecológico da região.

The Upper Juruá Extractive Reserve in the Brazilian Amazon: past and present

Abstract Amazonian livelihoods are largely dependent on rivers, with local protein consumption mainly relying on several species of fish. The UJER (Upper Juruá Extractive Reserve - Reserva Extrativista do Alto Juruá) is located in the state of Acre, bordering Peru and several indigenous areas. Here we summarize the data we collected in 1993/1994 on the population living along the banks of the Juruá, Tejo, Bagé, Igarapé São João and Breu rivers on crop cultivation, animal husbandry, and use of game and fish resources. We interviewed 133 individuals (94 on the Juruá and Tejo, 16 on Bagé, 16 on Igarapé São João and 7 on Breu rivers). Our results include a comprehensive description on local livelihoods, including the most important fish species for local subsistence considering gender and seasonality, the main husbandry and game species, and the items cultivated on the local agriculture. Whenever more recent information was available in the literature, we compared changes in livelihoods over time in the same region and also with the recent patterns observed in the Lower and in the Middle Juruá River. We hope to provide useful information to understand temporal changes in local livelihoods, which can help adapt and shape the ecological management in the region.

Resumo Comunidades da Amazônia são amplamente dependentes dos rios, com o consumo local de alimentos baseado em várias espécies de peixes. A UJER (Reserva Extrativista do Alto Juruá - Reserva Extrativista do Alto Juruá) está localizada no estado do Acre, na fronteira com o Peru e com várias áreas indígenas. Aqui resumimos os dados que coletamos em 1993/1994 da população que vive às margens dos rios Juruá, Tejo, Bagé, Igarapé São João e Breu sobre cultivo, criação de animais e uso de recursos de caça e peixes. Foram entrevistados 133 indivíduos (94 nos rios Juruá e Tejo, 16 em Bagé, 16 em Igarapé São João e 7 no rio Breu). Nossos resultados incluem uma descrição abrangente dos meios de subsistência locais, incluindo as espécies de peixes mais importantes para a subsistência local, considerando gênero e sazonalidade, bem como as principais espécies de criação, de caça e os itens cultivados na agricultura local., Quando foi possível obter informações da literatura, comparamos as mudanças nos meios de subsistência ao longo do tempo na mesma região e também com os padrões recentes observados no Baixo e no Médio Rio Juruá. Esperamos fornecer informações úteis para o entendimento das mudanças temporais nos meios de subsistência locais, o que pode ser útil ao manejo ecológico da região.

The Upper Juruá Extractive Reserve in the Brazilian Amazon: past and present / A Reserva Extrativista do Alto Juruá na Amazônia Brasileira: passado e presente

Amazonian livelihoods are largely dependent on rivers, with local protein consumption mainly relying on several species of fish. The UJER (Upper Juruá Extractive Reserve - Reserva Extrativista do Alto Juruá) is located in the state of Acre, bordering Peru and several indigenous areas. Here we summarize the data we collected in 1993/1994 on the population living along the banks of the Juruá, Tejo, Bagé, Igarapé São João and Breu rivers on crop cultivation, animal husbandry, and use of game and fish resources. We interviewed 133 individuals (94 on the Juruá and Tejo, 16 on Bagé, 16 on Igarapé São João and 7 on Breu rivers). Our results include a comprehensive description on local livelihoods, including the most important fish species for local subsistence considering gender and seasonality, the main husbandry and game species, and the items cultivated on the local agriculture. Whenever more recent information was available in the literature, we compared changes in livelihoods over time in the same region and also with the recent patterns observed in the Lower and in the Middle Juruá River. We hope to provide useful information to understand temporal changes in local livelihoods, which can help adapt and shape the ecological management in the region.

Comunidades da Amazônia são amplamente dependentes dos rios, com o consumo local de alimentos baseado em várias espécies de peixes. A UJER (Reserva Extrativista do Alto Juruá - Reserva Extrativista do Alto Juruá) está localizada no estado do Acre, na fronteira com o Peru e com várias áreas indígenas. Aqui resumimos os dados que coletamos em 1993/1994 da população que vive às margens dos rios Juruá, Tejo, Bagé, Igarapé São João e Breu sobre cultivo, criação de animais e uso de recursos de caça e peixes. Foram entrevistados 133 indivíduos (94 nos rios Juruá e Tejo, 16 em Bagé, 16 em Igarapé São João e 7 no rio Breu). Nossos resultados incluem uma descrição abrangente dos meios de subsistência locais, incluindo as espécies de peixes mais importantes para a subsistência local, considerando gênero e sazonalidade, bem como as principais espécies de criação, de caça e os itens cultivados na agricultura local. Quando foi possível obter informações da literatura, comparamos as mudanças nos meios de subsistência ao longo do tempo na mesma região e também com os padrões recentes observados no Baixo e no Médio Rio Juruá. Esperamos fornecer informações úteis para o entendimento das mudanças temporais nos meios de subsistência locais, o que pode ser útil ao manejo ecológico da região.

The Upper Juruá Extractive Reserve in the Brazilian Amazon: past and present†.

Amazonian livelihoods are largely dependent on rivers, with local protein consumption mainly relying on several species of fish. The UJER (Upper Juruá Extractive Reserve - Reserva Extrativista do Alto Juruá) is located in the state of Acre, bordering Peru and several indigenous areas. Here we summarize the data we collected in 1993/1994 on the population living along the banks of the Juruá, Tejo, Bagé, Igarapé São João and Breu rivers on crop cultivation, animal husbandry, and use of game and fish resources. We interviewed 133 individuals (94 on the Juruá and Tejo, 16 on Bagé, 16 on Igarapé São João and 7 on Breu rivers). Our results include a comprehensive description on local livelihoods, including the most important fish species for local subsistence considering gender and seasonality, the main husbandry and game species, and the items cultivated on the local agriculture. Whenever more recent information was available in the literature, we compared changes in livelihoods over time in the same region and also with the recent patterns observed in the Lower and in the Middle Juruá River. We hope to provide useful information to understand temporal changes in local livelihoods, which can help adapt and shape the ecological management in the region.

Controlling antimicrobial activity and drug loading capacity of chitosan-based layer-by-layer films.

We report on layer-by-layer (LbL) films of chitosans (CHI) and hyaluronic acid (HA) whose properties could be controlled by employing chitosans with different degrees of deacetylation (DD¯ ≈ 85%; 65%; 40%) and high average molecular weight (ca. 106 g/mol). In spite of their high molecular weight, these chitosans are soluble within a wide pH range, including physiological pH. HA/CHI LbL films produced from polymer solutions at pH 4.5 were thinner, smoother, more hydrophilic than those prepared at pH 7.2. This is attributed to the more extended conformation adopted by chitosan due to its very high charge density at low pH, favoring a compact chain packing during the film formation and resulting in lower film thickness and roughness. The smoother HA/CHI LbL films obtained at pH 4.5 were effective against Escherichia coli, while the thicker, rougher LbL films fabricated at pH 7.2 could be used in the controlled released of Rose Bengal dye. In summary, the tuning of only two parameters, i.e. solution pH and DD¯ of chitosans, provides access to a library of HA/CHI LbL films for tailored, diversified applications.

Epidemiologic analysis of salivary gland tumors over a 10-years period diagnosed in a northeast Brazilian population.

BACKGROUND: Salivary gland tumors (SGT) correspond to a heterogeneous group of lesions with variable biological behavior. The present study aimed to determine the distribution and demographic findings of salivary gland neoplasms in a northeast Brazilian population. MATERIAL AND METHODS: A retrospective descriptive cross-sectional study was performed. A total of 588 cases of SGT were diagnosed between 2006 and 2016 of 4 pathology services in the state of Sergipe, Brazil. All cases were reviewed, and data such as sex, age, anatomical location, and histopathological diagnosis were collected. RESULTS: A total of 470 (79.9%) tumors were benign and 118 (20.1%) were malignant. The majority of the patients were females (n=328, 55.8%) with an overall female:male ratio of 1.2:1. The major salivary glands were affected more than the minor glands (69.5% vs. 30.5%). Pleomorphic adenoma (n=419, 71.3%) and mucoepidermoid carcinoma (n=29, 4.9%) were the most frequent benign and malignant tumors, respectively. In addition, both benign and malignant tumors occurred more frequently in the parotid gland (n=300, 51%, p<0.05). CONCLUSIONS: The epidemiologic profile and clinical characteristics of SGT were similar to those described in other countries and other regions of Brazil. Epidemiological studies of SGT help to understand their clinical and pathological features and are essential to establish the proper management and prognosis.

Stress response pathways in the male germ cells and gametes.

The unfolded protein response (UPR) is a conserved and essential cellular pathway involved in protein quality control that is activated in response to several cellular stressors such as diseases states, ageing, infection and toxins. The cytosol, endoplasmic reticulum (ER) and mitochondria are continuously exposed to new proteins and in situations of aberrant protein folding; one of three lines of defence may be activated: (i) heat-shock response, (ii) mitochondrial UPR and (iii) ER UPR. These pathways lead to different signal transduction mechanisms that activate or upregulate transcription factors that, in turn, regulate genes that increase the cell's ability to correct the conformation of poorly folded proteins or, ultimately, lead to apoptosis. Despite the recent progress in understanding such biological processes, few studies have focused on the implications of the UPR in male infertility, highlighting the need for a first approach concerning the presence of these components in the male reproductive system. In testis, there is a high rate of protein synthesis, and the UPR mechanisms are well described. However, the presence of these mechanisms in spermatozoa, apparently transcriptionally inactive cells, is contentious, and it is unclear how sperm cells deal with stress. Here, we review current concepts and mechanisms of the UPR and highlight the relevance of these stress response pathways in male fertility, especially the presence and functional activation of those components in male germinal cells and spermatozoa.

Ultrastructural and metabolic disorders induced by short-term cadmium exposure in Avicennia schaueriana plants and its excretion through leaf salt glands.

Environmental cadmium (Cd) sources have increased in mangrove sediments in recent decades, inducing cellular damage to many plants. Avicennia schaueriana is abundant in mangrove sites and has been subject to Cd contamination. The possible effects of Cd toxicity and the structural and physiological disturbances to this plant were studied. Can this plant express early cellular tolerance mechanisms to such metal contamination? Seedlings of A. schaueriana were collected from sites of their natural occurrence, placed in plastic pots containing nutrient solution for 60 days, and subsequently exposed to increasing Cd concentrations for 5 days under experimental conditions. The anatomical, ultrastructural and physiological changes induced by Cd were analysed. Cd accumulated mainly in the root system and in pneumatophores, stems and leaves, induced differential accumulation of mineral nutrients, but did not induce necrosis or changes in leaf anatomy. However, there was a decrease in starch grains and an increase in deposited electron-dense material in the cortex and vascular bundles. Cd induced both increases in calcium (Ca) content in shoots and Ca oxalate crystal precipitation in leaf mesophyll and was detected in crystals and in the secretion of salt glands. Our observations and experimental results provide evidence of Cd tolerance in A. schaueriana. As a new feature, despite the clear cellular physiological disorders, this plant is able to eliminate Cd through leaf salt glands and immobilise it in Ca crystals, representing fast mechanisms for Cd exclusion and complexation in leaves in heavy metal coastal polluted marine ecosystems.

Isoform-specific GSK3A activity is negatively correlated with human sperm motility.

In mouse and bovine sperm, GSK3 activity is inversely proportional to motility. Targeted disruption of the GSK3A gene in testis results in normal spermatogenesis, but mature sperm present a reduced motility, rendering male mice infertile. On the other hand, GSK3B testis-specific KO is fertile. Yet in human sperm, an isoform-specific correlation between GSK3A and sperm motility was never established. In order to analyze GSK3 function in human sperm motility, normospermic and asthenozoospermic samples from adult males were used to correlate GSK3 expression and activity levels with human sperm motility profiles. Moreover, testicular and sperm GSK3 interactomes were identified using a yeast two-hybrid screen and coimmunoprecipitation, respectively. An extensive in-silico analysis of the GSK3 interactome was performed. The results proved that inhibited GSK3A (serine phosphorylated) presents a significant strong positive correlation (r = 0.822, P = 0.023) with the percentage of progressive human sperm, whereas inhibited GSK3B is not significantly correlated with sperm motility (r = 0.577, P = 0.175). The importance of GSK3 in human sperm motility was further reinforced by in-silico analysis of the GSK3 interactome, which revealed a high level of involvement of GSK3 interactors in sperm motility-related functions. The limitation of techniques used for GSK3 interactome identification can be a drawback, since none completely mimics the physiological environment. Our findings prove that human sperm motility relies on isoform-specific functions of GSK3A within this cell. Given the reported relevance of GSK3 protein-protein interactions in sperm motility, we hypothesized that they stand as potential targets for male contraceptive strategies based on sperm motility modulation.

Moving beyond classical markers of water quality: detection of enteric viruses and genotoxicity in water of the Sinos River.

It is well recognized that the classical biological and chemical markers of environmental pollution do not necessarily indicate the presence or absence of emerging threats to public health, such as waterborne viruses and genotoxicants. The purpose of this preliminary study was to evaluate the presence of material of enteroviruses (EV), rotavirus (RV) and adenovirus (AdV) and genotoxicity in water samples from points of routine monitoring of water quality in the main course of the Sinos River. The points are classified into different levels of pollution in accordance to the Brazilian federal regulations. Viral genomes from EV, AdV were detected in two of the 4 collection points regardless of the level of urbanisation of the surrounding areas. In contrast, genotoxicity was not observed in piava (Leporinus obtusidens) fingerlings cultivated on these same water samples. Results were compared with classical physical, chemical and microbiological parameters. There was no clear evidence of association between any of the classical markers and the presence of viral genomes in the water samples tested.

Three-dimensional finite element analysis of stress distribution in retention screws of different crown-implant ratios.

The retaining screw of the implant-supported dental prosthesis is the weakest point of the crown/implant system. Furthermore, crown height is another important factor that may increase the lever arm. Therefore, the aim of this study was to assess the stress distribution in implant prosthetic screws with different heights of the clinical crown of the prosthesis using the method of three-dimensional finite element analysis. Three models were created with implants (3.75 mm × 10 mm) and crowns (heights of 10, 12.5 and 15 mm). The results were visualised by means of von Mises stress maps that increased the crown heights. The screw structure exhibited higher levels of stresses in the oblique load. The oblique loading resulted in higher stress concentration when compared with the axial loading. It is concluded that the increase of the crown was damaging to the stress distribution on the screw, mainly in oblique loading.

Human adenovirus (HAdV), human enterovirus (hEV), and genogroup A rotavirus (GARV) in tap water in southern Brazil.

The effects of viral gastroenteritis are more devastating in children than in any other age category. Thus, children exposed to the consumption of low quality water are at an increased risk of infection, especially in regions where sanitation is inadequate. The present study aimed to provide a survey of the occurrence of representative enteric viruses: human adenovirus (HAdV), human enteroviruses (hEV), and genogroup A rotavirus (GARV) in tap water samples collected in public schools located at six municipalities of Rio Grande do Sul, southern Brazil. Seventy-three schools were included in the study and tap water samples were analyzed by conventional PCR for the presence of HAdV, hEV, and GARV genomes. hEV showed the highest detection rate (27.4%), followed by HAdV (23.3%), and GARV (16.4%). New approaches to water monitoring should be considered to promote a better water quality and reduce the risk of waterborne diseases, especially considering drinking water to be served to vulnerable individuals.

Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast.

The Infectious Bursal Disease Virus (IBDV) causes immunosuppression in young chickens. Advances in molecular virology and vaccines for IBDV have been achieved by viral reverse genetics (VRG). VRG for IBDV has undergone changes over time, however all strategies used to generate particles of IBDV involves multiple rounds of amplification and need of in vitro ligation and restriction sites. The aim of this research was to build the world's first VRG for IBDV by yeast-based homologous recombination; a more efficient, robust and simple process than cloning by in vitro ligation. The wild type IBDV (Wt-IBDV-Br) was isolated in Brazil and had its genome cloned in pJG-CMV-HDR vector by yeast-based homologous recombination. The clones were transfected into chicken embryo fibroblasts and the recovered virus (IC-IBDV-Br) showed genetic stability and similar phenotype to Wt-IBDV-Br, which were observed by nucleotide sequence, focus size/morphology and replication kinetics, respectively. Thus, IBDV reverse genetics by yeast-based homologous recombination provides tools to IBDV understanding and vaccines/viral vectors development.