Expression of LH receptor mRNA splice variants in bovine granulosa cells: Changes with follicle size and regulation by FSH in vitro

In cattle, most evidence suggests that granulosa cells express LH receptors (LHR) after (or as) the follicle becomes dominant, however there is some suggestion that granulosa cells from smaller pre-dominant follicles may express several LHR mRNA splice variants. The objective of this study was to measure LHR expression in bovine follicles of defined size and steroiclogenic ability, and in granulosa cells from small follicles (< 6 mm diameter) undergoing differentiation in vitro. Serniquantitative RT-PCR demonstrated that LHR mRNA was undetectable in granulosa cells of follicles < 7 mm diameter (nondominant follicles), and increased with follicle diameter in follicles > 7 mm diameter. Splice variants with deletions of exon 10 and part of exon 11 were detected as previously described, and we detected a novel splice variant with a deletion of exon 3. Cultured granulosa cells contained LHR mRNA, but with significantly greater amounts of variants with deletions of exon 10 and/or exon 11 compared with cells from dominant follicles. FSH increased the abundance of some but not all LHR mRNA splice variants in cultured granulosa cells. The addition of LH to cultured cells did not increase progesterone secretion, despite the presence of LHR mRNA. Collectively, these data suggest that granulosa cells do not acquire functional LHR until follicle dominance occurs.

Cytokine gene variants and venous thrombotic risk in the BRATROS (BRAZILIAN THROMBOSIS STUDY)

Introduction: Venous thrombosis (VT) and inflammation are two closely related entities. In the present investigation we assessed whether there is a relation between genetic modifiers of the inflammatory response and the risk of VT. Materials and methods: 420 consecutive and unrelated patients with an objective diagnosis of deep VT and 420 matched controls were investigated. The frequencies of the following gene polymorphisms were determined in all subjects: TNF-α- 308 G/A, LT-α+ 252 A/G, IL-6-174 G/C, IL1-ra 86 bp VNTR, IL-10-1082 A/G and CD-31 125 C/G. Results: Overall odds ratio (OR) for VT related to TNF-α- 308 G/A, LT-α+ 252 A/G, IL-6-174 G/C, A1 allele (4 bp repeat) of the IL1-ra 86 bp VNTR, IL-10-1082 A/G and CD-31 125 C/G were respectively: 1.0 (CI95: 0.8-1.5), 1.3 (CI95: 1.0-1.7), 1.1 (CI95: 0.9-1.5), 1.6 (CI95: 1-2.5), 1.2 (CI95: 0.8-1.7) and 0.8 (CI95: 0.6-1.1). A possible interaction between polymorphisms was observed only for the co-inheritance of the mutant alleles of the LT-α+ 252 A/G and IL-10-1082 G/A polymorphisms (OR = 2; CI95: 1.1-3.8). The risk of VT conferred by factor V Leiden and FII G20210A was not substantially altered by co-inheritance with any of the cytokine gene polymorphisms. Conclusions: Cytokine gene polymorphisms here investigated did not significantly influence venous thrombotic risk. © 2006 Elsevier Ltd. All rights reserved.

Detection of Streptococcus mutans and Streptococcus sobrinus in dental plaque samples from Brazilian preschool children by polymerase chain reaction

The purposes of this study were to detect S. mutans and S. sobrinus by polymerase chain reaction (PCR) amplification, and to relate their presence to the incidence of dental caries in 42 Brazilian preschool children. Dental plaque samples were collected from the cervical margin of all erupted teeth of 5-6 years old children with primary dentition, using a sterile explorer. Examination of the dmft (decayed, missing, filled teeth) index, performed following the World Health Organization (WHO) caries diagnostic criteria, showed a 2.71 score. Prevalence of S. mutans and S. sobrinus was respectively, of 85.7% and 14.3%; no dental plaque sample was either positive or negative for both bacterial species. Children harboring either S. mutans or S. sobrinus presented the same caries prevalence. PCR showed good discriminative ability for differentiation between these species, and suggested that it is a technique suitable for epidemiological studies on mutans streptococci.

Structural studies of Helicase NS3 variants from Hepatitis C virus genotype 3 in virological sustained responder and non-responder patients

Background. About 130 million people are infected with the hepatitis C virus (HCV) worldwide, but effective treatment options are not yet available. One of the most promising targets for antiviral therapy is nonstructural protein 3 (NS3). To identify possible changes in the structure of NS3 associated with virological sustained response or non-response of patients, a model was constructed for each helicase NS3 protein coding sequence. From this, the goal was to verify the interaction between helicases variants and their ligands. Findings. Evidence was found that the NS3 helicase portion of non-responder patients contained substitutions in its ATP and RNA binding sites. K210E substitution can cause an imbalance in the distribution of loads, leading to a decrease in the number of ligations between the essential amino acids required for the hydrolysis of ATP. W501R substitution causes an imbalance in the distribution of loads, leading and forcing the RNA to interact with the amino acid Thr269, but not preventing binding of ribavirin inhibitor. Conclusions. Useful information is provided on the genetic profiling of the HCV genotype 3, specifically the coding region of the NS3 protein, improving our understanding of the viral genome and the regions of its protein catalytic site. © 2010 Rahal et al; licensee BioMed Central Ltd.

Allelic variants of XRCC1 and XRCC3 repair genes and susceptibility of oral cancer in Brazilian patients

Background: The capacity for DNA repair is essential in maintaining cellular functions and homeostasis; however, this capacity can be altered based on DNA sequence variations in DNA repair genes, which may contribute to the onset of cancer. Many single-nucleotide polymorphisms (SNPs) in repair genes have been found to be associated with oral cancer. The aim of this study was to investigate the relationship between the presence of allelic variants Arg194Trp (rs:1799782) and Arg399Gln (rs: 25487) of XRCC1 gene and Thr241Met (rs: 861539) of XRCC3 gene and susceptibility to oral cancer. We also attempted to correlate the frequencies obtained for each of the SNPs to histopathological parameters. Methods: A case-control study was conducted with genomic DNA from 150 patients with oral squamous cell carcinomas and 150 controls. SNPs were genotyped by RFLP-PCR. Results: The presence of the polymorphic variants of the XRCC1 gene within codon 194 (OR 0.82, 95% CI: 0.44-1.51) and codon 399 (OR 0.94, 95% CI: 0.59-1.50) and within the XRCC3 gene (OR 0.72; 95% CI: 0.45-1.16) were not associated with an increased risk of oral cancer. A combinational analysis of SNPs in both genes indicated no association. The presence of the allelic variants of these two genes had no statistically significant effect on tumor differentiation, lymph node invasion or tumor size. Conclusions: These results suggest that allelic variants of XRCC1 and XRCC3 are not suitable markers for susceptibility to carcinomas of the oral cavity and are also not related to the later stages of such tumors. © 2012 John Wiley & Sons A/S.

A formação Irati (Grupo Passa Dois, Permiano, Bacia do Paraná) no furo de sondagem FP-01-PR (Sapopema, PR)

Pós-graduação em Geologia Regional - IGCE

Staining of esthetic brackets by plaque disclosing solutions

AIM: To evaluate the staining of esthetic orthodontic brackets by plaque disclosing solutions. METHODS: Two types of brackets manufactured by GAC/DENTSPLY(r) were evaluated: ceramic (n=30) and polycarbonate (n=30). The brackets were divided into 6 groups. Two control groups (n=6) were immersed in absolute ethanol: GI - ceramic brackets and GII - polycarbonate brackets. Four experimental groups (n=12) were immersed in different plaque disclosing solutions: GIII (ceramic brackets) and GIV (polycarbonate brackets) were immersed in Replak(r); GV (ceramic brackets) and GVI (polycarbonate brackets) were immersed in Replasul "S"(r). Relative quantitative analysis of the influence of plaque disclosing tablets on bracket staining was performed using reflectance spectrophotometry of stain deposition. Exploratory analysis of the data was performed using Analysis of Variance (ANOVA) in a 2x2 factorial setup (bracket x immersion) with additional treatments (controls). RESULTS: The results demonstrated that the ceramic brackets presented the highest amount of staining when Replasul "S"(r) was used (pd"0.05). However, when Replak(r) was used, no statistically significant difference was found in comparison with the control group (p>0.05). For polycarbonate brackets, staining was detected for both disclosing solutions (p>0.05). CONCLUSIONS: The disclosing solutions caused stain formation on polycarbonate brackets and, under the tested conditions, use of Replak(r) on ceramic brackets did not cause staining.

Transmission rate variation among three B chromosome variants in the fish Prochilodus lineatus (Characiformes, Prochilodontidae)

Estudos citogenéticos foram desenvolvidos em Prochilodus lineatus (Valenciennes 1836), os quais descrevem um interessante sistema de pequenos cromossomos supranumerários. O objetivo deste trabalho foi estudar a frequência e morfologia dos cromossomos B em indivíduos a partir da linhagem parental e os padrões de herança destes elementos em indivíduos obtidos a partir de cruzamentos controlados na espécie P. lineatus. A taxa de transmissão dos cromossomos B revelaram um kB=0,388 para o tipo acrocêntrico, kB=0,507 para o tipo metacêntrico e kB=0,526 submetacêntrico. Os resultados obtidos levantam hipóteses de que os cromossomos B do tipo acrocêntrico se encontram em fase de extinção, enquanto que os supranumerários do tipo metacêntrico e submetacêntrico encontram-se em fase de neutralização, seguindo uma taxa de transmissão Mendeliana.

Distribution of fluoride and calcium in plaque biofilms after the use of conventional and low-fluoride dentifrices

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

Plaque fluoride concentrations associated to the use of conventional and low-fluoride dentifrices

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

Plaque control improves the painful symptoms of oral lichen planus gingival lesions. A short-term study

BackgroundLichen planus is a mucocutaneous disease with manifestation in the oral mucosa, the gingiva being one of the most affected regions. In some cases, the lesion may be painful and lead to fragility of the tissues, so that precise diagnosis and adequate treatment are indispensible factors for improving the clinical condition. The aim of this study was to evaluate the effectiveness of plaque control in the improvement of clinical features and painful symptoms of oral lichen planus with gingival involvement.MethodsTwenty patients diagnosed with gingival lichen planus confirmed by histopathological examination were selected. The patients were evaluated by a trained examiner, with regard to the clinical features of the lesions [Index of Escudier etal. (Br J Dermatol, 157, 2007, 765)]; painful symptoms (Visual Analog Scale); and periodontally, as regards the visible plaque and gingival bleeding indices. Periodontal treatment consisted of supragingival scaling and oral hygiene instruction, with professional plaque removal afterward for a period of 4weeks. The entire sample was evaluated at the baseline and at the conclusion of treatment, and the results were analyzed by the Wilcoxon nonparametric test.ResultsThe data demonstrated that the majority of patients were women (90%), with a mean age of 55.9years. Periodontal treatment resulted in statistically significant reduction (P<0.05) in the periodontal indices, with consequent improvement in the clinical features and painful symptoms of the lesions.ConclusionsIt was demonstrated that plaque control was effective in improving the clinical features and painful symptoms of oral lichen planus with gingival involvement.

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.

The effect of chlorhexidine on plaque index and mutans streptococci in orthodontic patients: A pilot study

Aim: The purpose of this study was to assess chlorhexidine effects on plaque index and salivary levels of mutans streptococci (MS) when used as the immersion solution for removable orthodontic appliances and added to their acrylic resin composition. Methods: Forty-five patients (6 to 12 years old) were randomly assigned into three groups with 15 patients each. Group I (control)—without orthodontic appliances disinfection; Group II—removable orthodontic appliances which had been immersed in 0.12% chlorhexidine digluconate overnight (8 hours), and Group III—orthodontic appliances in which 0.12% chlorhexidine digluconate solution had been incorporated into their resin composition. Saliva was collected for quantification of MS and evaluation of plaque index was performed before and after installation of orthodontic appliance at 0, 2, 4, 6, 8, and 10 weeks. Data were analyzed by using analysis of variance. Results: Number of MS colonies in saliva and plaque index showed no statistically differences among groups at the different periods (p > 0.05). Conclusions: It could be concluded that chlorhexidine incorporation into the acrylic resin of removable orthodontic appliances at 0.12% concentration and immersion of the appliance into 0.12% chlorhexidine solution were not effective in reducing plaque index and the number of MS in saliva.

Venomics of the Australian eastern brown snake (Pseudonaja textilis): detection of new venom proteins and splicing variants

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