Field effect in oral precancer as assessed by DNA flow cytometry and array-CGH

J Oral Pathol Med (2012) 41 : 119–123 Objective: ‘Field cancerization’ is an accepted model for oral carcinogenesis. So far, genetically altered fields have been just reported in the presence of carcinomas. This study assessed the distant mirror fields (MFs) of oral precancer by DNA high‐resolution flow cytometry (hr DNA‐FCM) and array‐Comparative Genomic Hybridization (a‐CGH). Methods: Five leukoplakias without dysplasia (OLs), seven dysplastic leukoplakias (DOLs), and 12 corresponding visually normal and non‐dysplastic MFs were analyzed. DNA aneuploidy (DNA Index, DI ≠ 1) was detected by hr DNA‐FCM on DAPI stained nuclei suspensions. The epithelial DNA aneuploid subclones were FCM‐sorted to obtain genomic DNA for a‐CGH. Results: Mirror fields, OLs, and DOLs showed increasing prevalence of DNA aneuploidy of, respectively, 8%, 20%, and 57%. The average number of chromosome aberrations (Ch‐Abs) was 2.8 in MFs, 3 in OLs, and 10.6 in DOLs. MFs relative to OLs and DOLs had average numbers of Ch‐Abs, respectively, of 1.8 and 3.6. Ch‐Abs were also observed in DNA diploid sublines, and often the same aberrations were observed in both MFs and corresponding OLs/DOLs. Conclusion: DNA aneuploidy and Ch‐Abs in MFs, the last ones being mainly gains, indicate an early onset of field effect in oral carcinogenesis.     

Salivary shedding of Epstein–Barr virus and cytomegalovirus in people infected or not by human immunodeficiency virus 1

The purpose of this study is to determine the frequency of EBV and CMV DNA detection in saliva of HIV infected and non-HIV individuals and their siblings. The study group comprised 240 individuals. Group 1 comprised of 40 HIV-infected patients, group 2 40 non-HIV individuals, group 3 two siblings for each patient from group 1 (n = 80), and group 4 two siblings for each individual from group 2 (n = 80). Non-stimulated whole saliva was collected, DNA was extracted, and amplification was performed using a nested PCR protocol. EBV and CMV DNA was detected in 7/40 (17.5%) and 5/40 (12.5%) individuals from group 1, 8/40 (20%) and 3/40 (7.5%) from group 2, 11/80 (13.8%) and 2/80 (2.5%) from group 3, and 8/80 (10%) and 1/80 (1.3%) from group 4, respectively. Five (71.4%) out of seven HIV/EBV coinfected individuals of group 1 had a relative also infected with EBV (OR = 11.25, CI [1.75–72.5], p = 0.011). Regarding group 2, among the eight non-HIV and EBV-infected individuals, three (37.5%) had a relative also positive to EBV (p = 0.320). No individual HIV/CMV coinfected had a relative CMV infected (p = 1.00). Also, only one non-HIV and CMV-infected individual had a relative also positive to CMV (p = 0.075). EBV and CMV DNA was detected mainly in those who had HIV viral load counts <400/mL (71%, p = 0.2 and 100%, p = 1, respectively) and those who had CD4 T cells counts between 200 and 400/mm³ (57%, p = 0.544 and 60%, p = 0.249, respectively). HIV-infected individuals and healthy controls showed a similar frequency of viral DNA detection. EBV DNA was significantly amplified in saliva of household members of HIV/EBV coinfected individuals.     

Degree of conversion of nanofilled and microhybrid composite resins photo-activated by different generations of LEDs

Objective

This study aimed at evaluating the degree of conversion (DC) of four composite resins, being one nanofilled and 3 microhybrid resins, photo-activated with second- and third-generation light-emitting diodes (LEDs).

Material and methods

Filtek^TM Z350 nanofilled composite resins and Amelogen^® Plus, Vit-l-escence^TM and Opallis microhybrid resins were photo-activated with two second-generation LEDs (Radii-cal and Elipar Free Light^TM 2) and one third-generation LED (Ultra-Lume LED 5) by continuous light mode, and a quartz halogen-tungsten bulb (QHT, control). After 24 h of storage, the samples were pulverized into fine powder and 5 mg of each material were mixed with 100 mg of potassium bromide (KBr). After homogenization, they were pressed, which resulted in a pellet that was evaluated using an infrared spectromer (Nexus 470, Thermo Nicolet) equipped with TGS detector using diffuse reflectance (32 scans, resolution of 4 cm^-1) coupled to a computer. The percentage of unreacted carbon-carbon double bonds (% C=C) was determined from the ratio of absorbance intensities of aliphatic C=C (peak at 1637 cm-1) against internal standard before and after curing of the specimen: aromatic C-C (peak at 1610 cm-1).

Results

The ANOVA showed a significant effect on the interaction between the light-curing units (LCUs) and the composite resins (p<0.001). The Tukey''s test showed that the nanofilled resin (Filtek^TM Z350) and Opallis when photo-activated by the halogen lamp (QTH) had the lowest DC compared with the other microhybrid composite resins. The DC of the nanofilled resin (Filtek^TM Z350) was also lower using LEDs. The highest degrees of conversion were obtained using the third-generation LED and one of second-generation LEDs (Elipar Free Light^TM 2).

Conclusions

The nanofilled resin showed the lowest DC, and the Vit-l-escence^TM microhybrid composite resin showed the highest DC. Among the LCUs, it was not possible to establish an order, even though the second-generation LED Radii-cal provided the lowest DC.