Next-generation sequencing reveals large connected networks of intra-host HCV variants

Background: Next-generation sequencing (NGS) allows for sampling numerous viral variants from infected patients. This provides a novel opportunity to represent and study the mutational landscape of Hepatitis C Virus (HCV) within a single host.Results: Intra-host variants of the HCV E1/E2 region were extensively sampled from 58 chronically infected patients. After NGS error correction, the average number of reads and variants obtained from each sample were 3202 and 464, respectively. The distance between each pair of variants was calculated and networks were created for each patient, where each node is a variant and two nodes are connected by a link if the nucleotide distance between them is 1. The work focused on large components having > 5% of all reads, which in average account for 93.7% of all reads found in a patient. The distance between any two variants calculated over the component correlated strongly with nucleotide distances (r = 0.9499; p = 0.0001), a better correlation than the one obtained with Neighbour-Joining trees (r = 0.7624; p = 0.0001). In each patient, components were well separated, with the average distance between (6.53%) being 10 times greater than within each component (0.68%). The ratio of nonsynonymous to synonymous changes was calculated and some patients (6.9%) showed a mixture of networks under strong negative and positive selection. All components were robust to in silico stochastic sampling; even after randomly removing 85% of all reads, the largest connected component in the new subsample still involved 82.4% of remaining nodes. In vitro sampling showed that 93.02% of components present in the original sample were also found in experimental replicas, with 81.6% of reads found in both. When syringe-sharing transmission events were simulated, 91.2% of all simulated transmission events seeded all components present in the source.Conclusions: Most intra-host variants are organized into distinct single-mutation components that are: well separated from each other, represent genetic distances between viral variants, robust to sampling, reproducible and likely seeded during transmission events. Facilitated by NGS, large components offer a novel evolutionary framework for genetic analysis of intra-host viral populations and understanding transmission, immune escape and drug resistance.

Predictive maintenance of a worm reducer with an unbalanced load

Among all predictive maintenance techniques the oil analysis and vibration analysis are the most important for monitoring some mechanical systems. The integration of these techniques has potential to improve industrial maintenance practices and provide a better economic gain for industries. To study the integration of these two techniques, a test rig was set up to obtain an extreme working condition for the worm reducer used in this paper. The test rig was composed by a motor connected to a reducer through a flexible coupling and with an unbalanced load. The analysis of the results carried out by using a sample of the oil recommended by the manufacturer in extreme conditions, and using liquid contaminant is presented. From the results it was observed that if there is an abnormal instantaneous load in a system, the subsequent vibration analysis may not perceive what occurred if there was no permanent damage, which is not the case with the lubricant analysis.

Resistance training with excessive training load and insufficient recovery alters skeletal muscle mass-related protein expression

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Survival rate, load to fracture, and finite element analysis of incisors and canines restored with ceramic veneers having varied preparation design

Purpose: To evaluate the survival rate, success rate, load to fracture, and finite element analysis (FEA) of maxillary central incisors and canines restored using ceramic veneers and varying preparation designs.Methods and Materials: Thirty human maxillary central incisors and 30 canines were allocated to the following four groups (n=15) based on the preparation design and type of tooth: Gr1 = central incisor with a conservative preparation; Gr2 = central incisor with a conventional preparation with palatal chamfer; Gr3 = canine with a conservative preparation; Gr4 = canine with a conventional preparation with palatal chamfer. Ceramic veneers (lithium disilicate) were fabricated and adhesively cemented (Variolink Veneer). The specimens were subjected to 4 x 106 mechanical cycles and evaluated at every 500,000 cycles to detect failures. Specimens that survived were subjected to a load to fracture test. Bidimensional models were modeled (Rhinoceros 4.0) and evaluated (MSC.Patrans 2005r2 and MSC.Marc 2005r2) on the basis of their maximum principal stress (MPS) values. Survival rate values were analyzed using the Kaplan-Meier test (alpha = 0.05) and load to fracture values were analyzed using the Student t-test (alpha = 0.05).Results: All groups showed 100% survival rates. The Student t-test did not show any difference between the groups for load to fracture. FEA showed higher MPS values in the specimens restored using veneers with conventional preparation design with palatal chamfer.Conclusion: Preparation design did not affect the fracture load of canines and central incisors, but the veneers with conventional preparation design with palatal chamfer exhibited a tendency to generate higher MPS values.

Diagnostic tests for hepatitis C: Recent trends in electrochemical immunosensor and genosensor analysis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of thermo-mechanical load cycling on silorane-based composite restorations

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Salivary cortisol response to concurrent strength and high-intensity aerobic exercise: a pilot study

Aim. The aim of this pilot study was to compare strength performance and salivary cortisol levels response during a single strength session, and a strength session after 30 min of high-intensity aerobic exercise (concurrent condition).Methods. Saliva was collected from 7 male subjects, before and after all exercise bouts, and the maximum number of repetitions (MNR) and total volume (TV) in the different conditions assessed.Results. The MNR and TV were reduced in concurrent condition compared with control condition. Strength exercise in the concurrent condition induced higher salivary cortisol in relation to strength exercise or to and high-intensity aerobic exercise, separately.Conclusion. The different salivary cortisol profile in response to concurrent exercise reflects the faster reactivity of the hypothalamic-pituitary-adrenocortical system in this circumstance.

Cytochemical study of Rhodnius neglectus and Rhodnius prolixus salivary gland cells (Hemiptera, Triatominae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Salivary and serum proteomics in head and neck carcinomas before and after surgery and radiotherapy

Several body fluids have been evaluated as new sources for cancer biomarker discovery. In this context, salivary and serum proteomics seem promising diagnostic and predictive tools for head and neck diseases. In the present study, we performed a proteomic analysis of saliva and serum from patients presenting head and neck squamous cell carcinoma (HNSCC) and compared the results before and after therapy. In saliva of cancer patients, we observed an altered protein profile, including over-expression of PLUNC and zinc-alpha-2-glycoprotein. Both proteins may contribute to control tumor growth and, therefore, represent targets for new analysis. We also detected serotransferrin and a modified transthyretin form with altered levels in serum from patients. Comparing preoperative and postreatment samples, the results showed that the protein profile after treatment reverted to a pattern closer to those observed for controls. These results add information on the role of secreted proteins in the cancer process and emphasize the potential of saliva and serum analysis for diagnosis and monitoring of HNSCC.

Use of salivary biomarkers for diagnosis of periodontal disease activity: a literature review

Periodontal disease is an infectious disease characterized by the connective tissue destruction and consequent alveolar bone loss in response to plaque accumulation on the tooth surface. The clinical diagnosis of periodontal disease is based both on clinical examination involving the evaluation of probing depth and radiographic examination of alveolar bone loss but these examinations are not enough to determine the activity of the disease process. For that reason, it has been proposed to seek predictive disease markers in an attempt to assess the disease activity and so, evaluate the efficacy of the periodontal disease treatment. The aim of this review is to present recent advances in the development of proteomic, genomics and microbial biomarkers and potential clinical applications. It was concluded that periodontal treatment based on assessing the levels of salivary biomarkers emerges as a promising method in near future and will become an integral part of the evaluation of periodontal health.

Influence of standard load micro and nanopatterned in surface roughness of bleached teeth and submitted to different surface treatments?

The aim of this study was evaluate the dental enamel after whitening treatment with Opalescence Boost PF® 38%, correlating the structural alterations in the surface of the enamel with its respective pH and verify if whitened teeth submitted to different finishing and polishing techniques show similar surface texture to healthy teeth (control group). Sixty premolars were divided in 6 groups (n = 10), which had been immersed in artificial saliva during all the experiment. Protocol whitening was performed according to the manufacturer recommendations, and then the specimens were submitted to different polishing technique with Sof-Lex Pop On® disks, Flex Diamond® felt disks using two different micrometric polishing pastes (Enamelize® and Diamond Polish®) and two nanometric polishing pastes (Lummina-E Diamond and Lummina-E Alumina), according to the groups. Representative specimens were analyzed in scanning electronic microscopy (SEM). Whitening gel used in this experiment had modified the morphologic aspect of the enamel surface. It was found that two nanometric polishing pastes (G5 and G6) promoted a less rough surface compared to control group even after the whitening process.

Action of the salivary microorganisms having sorbitol as substratum

Microorganisms living in the saliva are able to ferment some sweetener types and to promote the fall of pH that has its critical point around 5.5; in this situation, they equal the input and the calcium output in the processes of remineralization and demineralization. For iteration of the three essential factors to the installation of the dental caries (microorganism, host and diet) suggested the accomplishment of a work that involved one of these factors. This work aims to study this issue, through the indication of substitute sweetener of the saccharose, whenever possible, it solved to evaluate pH of the saliva, exposed to sorbitol.

Platelets can be a biological compartment for the Hepatitis C Virus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Load-application devices: a comparative strain gauge analysis

In view of the low loading values commonly employed in dentistry, a load-application device (LAD) was developed as option to the universal testing machine (UTM), using strain gauge analysis. The aim of this study was to develop a load-application device (LAD) and compare the LAD with the UTM apparatus under axial and non-axial loads. An external hexagonal implant was inserted into a polyurethane block and one EsthetiCone abutment was connected to the implant. A plastic prosthetic cylinder was screwed onto the abutment and a conical pattern crown was fabricated using acrylic resin. An impression was made and ten identical standard acrylic resin patterns were obtained from the crown impression, which were cast in nickel-chromium alloy (n=10). Four strain gauges were bonded diametrically around the implant. The specimens were subjected to central (C) and lateral (L) axial loads of 30 kgf, on both devices: G1: LAD/C; G2: LAD/L; G3: UTM/C; G4: UTM/L. The data (με) were statistically analyzed by repeated measures ANOVA and Tukey's test (p<0.05). No statistically significant difference was found between the UTM and LAD devices, regardless of the type of load. It was concluded that the LAD is a reliable alternative, which induces microstrains to implants similar to those obtained with the UTM.

Immunization of Wistar female rats with 255-Gy-irradiated Toxoplasma gondii: Tissue parasitic load and lactogenic quantification

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)