Effect of low-level laser therapy on the gene expression of collagen and vascular endothelial growth factor in a culture of fibroblast cells in mice.

Low-level laser therapy treatment (LLLT) is widely used in rehabilitation clinics with the aim of accelerating the process of tissue repair; however, the molecular bases of the effect of LLLT have not been fully established. The aim of the present study was to evaluate the influence of the exposure of different doses of LLLT on the expression of collagen genes type I alpha 1 (COL1α1) and vascular endothelial growth factor (VEGF) in the fibroblast cells of mice (L929) cultivated in vitro. Fibroblast cells were irradiated with a Gallium-Arsenide laser (904 nm) every 24 h for 2 consecutive days, stored in an oven at 37 °C, with 5% CO2 and divided into 3 groups: G1-control group, G2-irradiated at 2 J/cm(2), and G3-irradiated at 3 J/cm(2). After irradiation, the total RNA was extracted and used in the complementary DNA (cDNA) synthesis. The gene expression was analyzed by real-time polymerase chain reaction. The cells irradiated in G2 exhibited a statistically significant growth of 1.78 in the expression of the messenger RNA (mRNA) of the COL1α1 gene (p = 0.036) in comparison with G1 and G3. As for the VEGF gene, an increase in expression was observed in the two irradiated groups in comparison with the control group. There was an increase in expression in G2 of 2.054 and G3 of 2.562 (p = 0.037) for this gene. LLLT (904 nm) had an influence on the expression of the genes COL1α1 (2 J/cm(2)) and VEGF (2 e 3 J/cm(2)) in a culture of the fibroblast cells of mice.

Influence of different photo-activation distances on cytotoxicity of a dental adhesive model resin.

AIM: This study evaluated the cytotoxicity of a dental bonding model resin (DBMR) submitted to different photo-activation distances. METHODS: A monomer mixture based on Bis-GMA and HEMA was used to assess the cytotoxicity in a mouse fibroblast-cell line. To promote different photo-activation distances glass slides were interposed between DBMR surface and halogen light curing unit (LCU) tip. Afterwards, the specimens were immersed in RPMI culture medium for 24 h to obtain extracts. The extracts were incubated in contact with the cells for 24 h. Finally, an MTT colorimetric assay was used to assess the cytotoxicity. The cell viability data (absorbance) were analyzed by one way ANOVA followed by Tukey's test (P<0.05). RESULTS: The light output decreased according to the increase in the number of glass slides between the halogen LCU tip and DBMR surface. Yet, the distance between the tip of the curing light system and the specimens had significant influence on the cytotoxicity. All extracts produced by groups submitted to different photo-activation distances showed cytotoxic effect after 24h of incubation. CONCLUSION: The photo-activation distance and the interposition of glass slides between LCU tip and DBMR was shown to play an important role in the cytotoxic effect.

Polimorfismos genéticos da kappa-caseína e da beta-lactoglobulina e produção de leite em bovinos / Kappa-casein and beta-lactoglobulin genetic polymorphisms and milk yield in bovines

As variantes gênicas da beta-lactoglobulina (β-LG) e da kappa-caseína (κ-CN) bovinas são associadas à produção, qualidade e características de processamento do leite. O objetivo deste trabalho foi analisar as frequências dos genótipos AA, AB e BB, por meio da técnica de PCR-RFLP, da β-LG e da κ-CN bovinas, e suas associações com a produção de leite (kg leite/dia) em bovinos das raças Girolanda, Holandesa e Jersey. Para a κ-CN, a frequência do genótipo AA foi maior nos animais das raças Holandesa (37%) e Girolanda (63%). Na raça Jersey, houve predomínio do genótipo BB (60%). Para a β-LG, o genótipo AB foi o mais encontrado nas raças Girolanda (54%) e Holandesa (58%), enquanto nos animais da raça Jersey houve predomínio do genótipo BB (45%). Houve associação do alelo B da κ-CN com maior produtividade leiteira nas raças Girolanda e Holandesa, e do alelo A da β-LG com maior produtividade de leite na raça Jersey. As variantes genéticas da κ-CN podem ser usadas como marcadores na seleção para a produtividade leiteira nas raças Girolanda e Holandesa. Para a raça Jersey, as variantes da β-LG seriam mais adequadas para essa seleção.

Bovine beta-lactoglobulin (β-LG) and kappa-casein (κ-CN) genic variants are associated with productivity, quality and processing features of milk. The objective of this study was to analyze through the PCR-RFLP technique, the frequency of AA, AB and BB genotypes of bovine β-LG and κ-CN, and their association to milk production (kg milk/day) in Girolanda, Holstein and Jersey cattle. For κ-CN, the frequency of the AA genotype was higher in Holstein (37%) and Girolanda (63%), while there was a predominance of the BB genotype in Jersey (60%). For β-LG, the BB genotype was the most found in Girolanda (54%) and Holstein (58%), while there was a predominance of the BB genotype (45%) in Jersey. There was a positive association between B allele of κ-CN and milk production in the Girolanda and Holstein cattle and between A allele of β-LG and milk production in the Jersey cattle. Genetic variants of κ-CN could be used as markers for the selection for productivity in Girolanda and Holstein cattle. The genetic variants of β-LG would be more appropriate for this selection in the Jersey breed.

Association of dental caries with HLA Class II allele in Brazilian adolescents.

The purpose of this study was to evaluate the association between the frequencies of DR and DQ HLA class II alleles and the prevalence of dental caries in Brazilian adolescents. The study sample consisted of 164 adolescents aged 15- 19 years. For the assessment of caries experience the DMFT index was used, according to World Health Organization criteria. DNA samples of the adolescents were extracted from oral mucosa cells. The amplification of the alleles HLA-DR and HLA-DQ was carried out by polymerase chain reaction with sequence-specific primers (PCR-SSP). The products of PCR-SSP were electrophoresed and analyzed in agarose gel. Statistical analysis was performed using simple and multiple logistic regression to test the association between the adolescents' caries experience and the presence of HLA-DR and -DQ alleles. Sociodemographic covariates were included in the model. The significance level for all analyses was p < 0.05. The prevalence of caries was 60.4% and the mean DMFT was 2.41 ± 2.53. Adolescents positive for HLA-DQ2 allele were less likely to have dental caries than their counterparts who were negative for this allele (OR = 0.33, CI 0.16-0.66), and those whose mothers presented low levels of education were 2.01 (CI 1.02-3.97) times more likely to have dental caries. There was no statistical difference between HLA-DR4, -DQ4, -DQ5, -DQ6 and dental caries. Our data provide evidence that genes within the MHC, especially the DQ2 group, may influence susceptibility to dental caries in Brazilian adolescents.

Genotypic Diversity of Streptococcus mutans in Caries-Free and Caries-Active Preschool Children.

Aim. The aim of the present paper was to evaluate the genotypic diversity of S. mutans in caries-free and caries-active preschool children in Brazil. Design. Twenty-eight preschool children were examined regarding caries experience by the dmft index. DNA from 280 isolates of S. mutans was extracted. S. mutans evaluated using to the PCR method, with primers for the glucosyltransferase gene. The genetic diversity of S. mutans isolates was analyzed by arbitrary primed-PCR (AP-PCR) reactions. The differences between the diversity genotypic and dmft/caries experience were evaluated by chi2 test and Spearman's correlation. Results. The Spearman correlation test showed a strong association between genotypic diversity and caries experience (r = 0.72; P < .001). There were more S. mutans genotypes in the group of preschool children with dental caries, compared with the caries-free group. Among the children with more than 1 genotype, 13 had dental caries (2 to 5 genotypes) and 4 were caries-free (only 2 genotypes). Conclusion. Our results support the previous findings of genetic diversity of S. mutans in preschool children being associated with dental caries. The investigation of such populations may be important for directing the development of programs for caries prevention worldwide.

Chromosome 22q a frequent site of allele loss in head and neck carcinoma.

BACKGROUND: Loss of heterozygosity (LOH) correlates with inactivated tumor suppressor genes. LOH at chromosome arm 22q has been found in a variety of human neoplasms, suggesting that this region contains a tumor suppressor gene(s) other than NF2 important to tumorigenesis. The aim of this study was to evaluate the presence of LOH on chromosome 22q11.2-13 and determine whether there was a relationship between loss in this genomic region and tumor histologic parameters, anatomic site, and survival in patients with squamous cell carcinoma of the head and neck (HNSCC). METHODS: Fifty matched blood and HNSCC tumor samples taken at the time of surgical treatment were evaluated for LOH by use of four microsatellite markers mapping to 22q11.2-q13. Clinical information was available for all patients. The frequency and distribution of LOH was correlated with clinical (age, sex, use of tobacco and alcohol, site of primary tumor, clinical stage, adjuvant therapy and overall survival) and histologic parameters (histopathologic stage, tumor differentiation). RESULTS: LOH at 22q was found in 19 of 50 (38%) informative tumors. The respective incidence of allelic loss for the patients was as follows: 28% at D22S421, 10% at D22S277, 8% at D22S446, and 4% at D22S280. No statistical differences were apparent with a mean follow-up of 30 months. Laryngeal tumors showed a higher incidence of LOH compared with oral tumors. CONCLUSIONS: These results suggest that the D22S277 locus may be closely linked to a tumor suppressor gene (TSG) and involved in upper aerodigestive tract carcinogenesis. In particular, laryngeal tumors may harbor another putative TSG on 22q11.2-q12.3 that may play a role in aggressive stage III/IV disease.

Comparative genomic hybridization analysis detects frequent over-representation of DNA sequences at 3q, 7p, and 8q in head and neck carcinomas.

Comparative genomic hybridization (CGH) was used to identify chromosomal imbalances in 19 samples of squamous cell carcinoma of the head and neck (HNSCC). The chromosome arms most often over-represented were 3q (48%), 8q (42%), and 7p (32%); in many cases, these changes were observed at high copy number. Other commonly over-represented sites were 1q, 2q, 6p, 6q, and 18q. The most frequently under-represented segments were 3p and 22q. Loss of heterozygosity of two polymorphic microsatellite loci from chromosome 22 was observed in two tongue tumors, in agreement with the CGH analysis. Gains of 1q and 2q material were detected in patients exhibiting a clinical history of recurrence and/or metastasis followed by terminal disease. This association suggests that gain of 1q and 2q may be a new marker of head and neck tumors with a refractory clinical response.