The differential roles of D-Pax2 variants in regulating Drosophila eye and bristle development

The ability to appropriately interact with the environment is crucial to an organism’s survival. The establishment of functional sensory systems, such as the bristles and eyes in Drosophila, is a critical event during the development of the organism. The transcription factor D Pax2 is involved in the differentiation of the shaft and glial cells in the developing bristle (Kavaler et al., Dev, 126:2261-2272, 1999) and of the cone and primary pigment cells in the developing eye (Fu and Noll, Genes Dev, 11:389-405, 1997). How D-Pax2 contributes to distinct differentiative pathways in different cell types is not known. Recent work by Anna Czechowski and Katherine Harmon (personal communication) identified a mutation in the D-Pax2 gene that introduced a stop codon at the end of exon 9, effectively truncating the protein. This mutation affects bristle, but not eye, development. We thus suspected regions after exon 9 are required for D-Pax2 function only in the bristles and may also be associated with alternative splicing of the D Pax2 transcript. We plan to assess the role of the carboxy terminal region of the protein by establishing transgenic lines bearing rescue constructs of D-Pax2 with either the complete coding sequence or with deletions of specific exons. To date, we have generated the first rescue construct bearing the complete coding region of the gene driven by a 3 KB upstream regulatory region of D-Pax2 and are currently generating transgenic fly lines with this construct.

Splice variants of the human ZC3H14 gene generate multiple isoforms of a zinc finger polyadenosine RNA binding protein

The human ZC3H14 gene encodes an evolutionarily conserved Cys(3)His zinc finger protein that binds specifically to polyadenosine RNA and is thus postulated to modulate post-transcriptional gene expression. Expressed sequence tag (EST) data predicts multiple splice variants of both human and mouse ZC3H14. Analysis of ZC3H14 expression in both human cell lines and mouse tissues confirms the presence of multiple alternatively spliced transcripts. Although all of these transcripts encode protein isoforms that contain the conserved C-terminal zinc finger domain, suggesting that they could all bind to polyadenosine RNA, they differ in other functionally important domains. Most of the alternative transcripts encode closely related proteins (termed isoforms 1, 2. 3, and 3short) that differ primarily in the inclusion of three small exons, 9, 10, and 11, resulting in predicted protein isoforms ranging from 82 to 64 kDa. Each of these closely related isoforms contains predicted classical nuclear localization signals (cNLS) within exons 7 and 11. Consistent with the presence of these putative nuclear targeting signals, these ZC3H14 isoforms are all localized to the nucleus. In contrast, an additional transcript encodes a smaller protein (34 kDa) with an alternative first exon (isoform, 4). Consistent with the absence of the predicted cNLS motifs located in exons 7 and 11, ZC3H14 isoform 4 is localized to the cytoplasm. Both EST data and experimental data suggest that this variant is enriched in testes and brain. Using an antibody that detects endogenous ZC3H14 isoforms 1-3 reveals localization of these isoforms to nuclear speckles. These speckles co-localize with the splicing factor, SC35, suggesting a role for nuclear ZC3H14 in mRNA processing. Taken together, these results demonstrate that multiple transcripts encoding several ZC3H14 isoforms exist in vivo. Both nuclear and cytoplasmic ZC3H14 isoforms could have distinct effects on gene expression mediated by the common Cys(3)His zinc finger polyadenosine RNA binding domain. (C) 2009 Elsevier B.V. All rights reserved.

Wild rabies virus detection by plaque assay from naturally infected brains in different species

A simple, sensitive and specific plaque assay protocol for the detection of wild type rabies virus in different species is described using confluent monolayers of chicken embryo cells in 6-well plates. Plaques are produced after application of either agarose or Sephadex G-100 overlay onto cell monolayers and incubation for 96 h after virus infection at 37 degreesC. The parameters affecting plaque appearance include cell seeding concentration, overlay composition and time of incubation after infection. Optimal conditions are seeding at a concentration of 4 x 10(6) cell/cm(3), incubation at 37 degreesC in 5% CO2 atmosphere during 96 h, using either 1% agarose or 2% Sephadex G-100 overlays. The described plaque assay would be a new valuable too] in conducting various quantitative investigations, since the chicken embryo cells are susceptible to rabies virus infection from all species studied. (C) 2004 Elsevier B.V. All rights reserved.

Effects of Regular and Low-fluoride Dentifrices on Plaque Fluoride

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

The Effect of Different Fluoride Concentrations and pH of Dentifrices on Plaque and Nail Fluoride Levels in Young Children

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

Effect of Calcium Pre-rinse and Fluoride Dentifrice on Enamel and on Dental Plaque Formed In Situ

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

Presence of mutans streptococci and Candida spp. in dental plaque/dentine of carious teeth and early childhood caries

This study determined the presence of mutans streptococci and Candida spp. in supragingival. dental plaque and infected dentine of caries-free children, with early childhood caries and caries. Pooled samples of dental plaque and infected dentine were collected from 56 children aged 1-5 years, which were divided into 3 groups: early childhood caries (ECC); caries and caries-free. Infected dentine was collected in ECC and caries groups to compare the frequency of these microorganisms in the collected sites. The samples were inoculated in SB20 and SA medium, for mutans streptococci and Candida spp., respectively, and incubated at 37 degrees C for 48 h. Colony growth was verified and the identification was performed by biochemical tests and CHROMagar Candida. Fisher's test or chi-square (chi(2)) were applied (p = 0.05). The more prevalent species were S. mutans and Candida albicans in ECC (85.4% and 60.4%, respectively), independently of the sample site. S. mutans only was significantly associated with carious teeth, whether in early childhood caries or not. However, the frequency of C. albicans in ECC was higher when compared to caries and caries-free groups. There is a significant association between the presence of C. albicans and early childhood caries. (c) 2006 Elsevier Ltd. All rights reserved.

The Effect of a Supragingival Plaque-Control Regimen on the Subgingival Microbiota in Smokers and Never-Smokers: Evaluation by Real-Time Polymerase Chain Reaction

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

The antibacterial effect of photodynamic therapy in dental plaque-derived biofilms

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effects of ultrasonic, electric, and manual toothbrushes on subgingival plaque composition in orthodontically banded molars

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

Oral Hygiene Indirect Instruction and Periodic Reinforcements: Effects on Index Plaque in Schoolchildren

The aim of this study was to evaluate the effectiveness of the indirect instruction and the influence of the periodic reinforcement on the plaque index in schoolchildren. Forty schoolchildren aged from 7 to 9 years old were selected from a public school. After determining the initial O'Leary Plaque Index all schoolchildren were submitted to a program for oral hygiene through indirect instruction - "The Smiling Robot". The schoolchildren were divided into 2 groups: with and without motivation reinforcement. The index plaque exam was performed in both groups after 30, 60 and 90 days of the educational program. Comparing the groups, the plaque index decreasing could be observed in the group with reinforcement with statistically significant difference. For the group with reinforcement, statistically significant difference among the evaluations was found. For the group without reinforcement, significant decrease in the plaque index was found after 30 days when compared to the first, third and fourth evaluations. The indirect instruction with "The Smiling Robot "promoted a positive initial impact on the decrease of plaque index in the schoolchildren. The periodic reinforcements showed snore suitable results and significant reduction of the plaque index in the course of the evaluations.