Are salivary NO2- / NO2- and NO3- levels biomarkers for dental caries in children? Systematic review and meta-analysis.

The literature is conflicting regarding salivary nitrite (NO2-)/nitrite and nitrate (NO2- and NO3-) levels in children affected by dental caries. For this reason, a systematic review to provide a consensus on the subject was propose, whose objective is to verify whether these molecules could be used as biomarkers in children with caries. A comprehensive search was performed on online database and eleven articles were included in the meta-analysis. The methodological quality of studies was assessed by Newcastle-Ottawa Scale recommended for case-control studies and by AXIS tool for cross-sectional studies. Grading of Recommendations Assessment, Development and Evaluation was used for the assessment of the certainty of the evidence for each outcome. The results showed lower NO2- levels in the group of children affected by dental caries (SMD = -2.18 [-3.24, -1.13], p < 0.01). Age, saliva collection and methods of evaluation can impact the results. When evaluating the severity of the condition, an important variation was detected in relation to the different evaluation methods NO2-/NO2- and NO3-. In conclusion, based on the evidence presented, the results suggest that NO2- levels in saliva are a possible biomarker of dental caries. Results should be evaluated with caution due to the very low evidence from primary studies. Longitudinal studies are necessary to strengthen this hypothesis.

The effect of toothpaste with reduced concentration of fluoride-containing sodium trimetaphosphate and polyols on initial enamel erosion.

OBJECTIVES: This in vitro study evaluated the efficacy of toothpaste containing fluoride (F), sodium trimetaphosphate (TMP), and xylitol and erythritol (XE) to inhibit or repair initial enamel erosion lesions. MATERIALS AND METHODS: Bovine enamel blocks (n = 120) were selected according to surface hardness (SH) and randomly divided into 5 experimental groups (n = 24 blocks/group): Placebo (no F, TMP, XE); 1100 ppm F; 16% xylitol + 4% erythritol (XE); 200 ppm F + 0.2% TMP (200 ppm F/TMP); and 200 ppm F + 0.2% TMP + 16% xylitol + 4% erythritol (200 ppm F/TMP/XE). The sound and softened blocks were immersed in toothpaste slurry in human saliva for 2 min. The blocks were then submitted to 4 erosive challenges in citric acid (0.75%, pH 3.5), each challenge for 1 min, with stirring. The SH of the blocks was determined after treatment (t) and after the 4 erosive challenges. In addition, the precipitates were analyzed by scanning electron microscopy (SEM). Variables were submitted to a two-way repeated-measures analysis of variance followed by a Student-Newman-Keuls test (p < 0.05). RESULTS: Toothpaste containing 200 ppm F/TMP/XE led to the highest protective and repair effect compared to the other groups (p < 0.001). The protective and repair effect was XE > 200 ppm F/TMP > 1100 ppm F > placebo (p < 0.001). All groups produced precipitates with a thicker layer for XE and TMP groups. CONCLUSIONS: Toothpaste containing 200 ppm F, TMP, and polyols demonstrated a superior protective and repair effect in initial enamel erosive lesions in vitro. CLINICAL RELEVANCE: Toothpaste containing F and polyols could be an advantage in patients affected by dental erosion, due to its higher preventive potential, mainly in individuals who frequently drink acidic beverages. However, future studies are needed to confirm these results.

Combined effect of casein phosphopeptide-amorphous calcium phosphate and sodium trimetaphosphate on the prevention of enamel demineralization and dental caries: an in vitro study.

OBJECTIVE: To evaluate the effect of treatment with fluoridated toothpaste supplemented with a combination of sodium trimetaphosphate (TMP) and casein phosphopeptide-amorphous calcium phosphate (MI Paste Plus®) on the demineralization of dental enamel. METHODS: Bovine enamel blocks selected by initial surface hardness (SHi) were randomly allocated into six groups (n = 12), according to the test toothpastes: (1) without F-TMP-MI Paste Plus® (Placebo); (2) 1100 ppm F (1100F); (3) MI Paste Plus®; (4) 1100F + MI Paste Plus® (1100F-MI Paste Plus), (5) 1100F + 3 % TMP (1100F-TMP); and (6) 1100F-TMP + MI Paste Plus® (1100F-TMP-MI Paste Plus). Blocks were treated two times per day with slurries of toothpaste (1 min), and groups 4 and 6 received an application of MI Paste Plus (3 min). Next, the samples were subjected to five pH cycles (demineralizing/remineralizing solutions) at 37 °C, to produce subsurface enamel lesions.Thereafter, the blocks were maintained for 2 days in fresh remineralizing solution. After pH cycling, the following were obtained: percentage of surface hardness loss (%SH); integrated loss of subsurface hardness (ΔKHN); profile analysis and lesion depth subsurface through polarized light microscopy (PLM); scanning electron microscopy (SEM); and fluoride (F), calcium (Ca), and phosphorus (P) in the enamel. The data were subjected to ANOVA (1-criterion), followed by the Student-Newman-Keuls test (p < 0.001). RESULTS: The 1100F-TMP-MI Paste Plus group showed better results for SHR, ΔKHN, and PLM (p < 0.001). The F concentration was similar among all groups (p > 0.001). The 1100F-TMP-MI Paste Plus group showed the highest concentration of Ca and P in the enamel (p < 0.001). CONCLUSION: The application of 1100F-TMP-MI Paste Plus promoted a higher inhibitory effect against enamel demineralization. CLINICAL SIGNIFICANCE: The combination of treatments with F, TMP, and MI Paste Plus® can be an effective alternative to improve the oral health of individuals, especially those with high activity of dental caries and at high risk for its development.

Retrospective Review of Oral Probiotic Therapy.

Purpose: There have been many in vitro studies reporting on the efficacy of probiotic bacteria in inhibiting pathogens, and there have been published studies reporting on the inhibitor effects of probiotic bacteria on the salivary levels of bacterial pathogens. However, there have not been but a few studies on the clinical benefits of oral probiotic therapy. Study design: Dental records of 60 patients that were enrolled in an Institutional Review Board approved study were reviewed as to current caries activity status with measurement of the Decayed Missing Filled Teeth index and by Caries Management By Risk Assessment (CAMBRA) determination. The current oral health status was compared to the prior-to-study enrollment status and then analyzed in respect to published national norms. The data (without any identifiers) had a statistical analysis by a blinded biostatistician. The data was subjected to statistical analysis (Statsgraphic) before and after the probiotic therapy. Results: Of the 53 subjects available for follow up, only 4 had remained caries active with a grand total of 27 carious lesions being detected and subsequently restored in this group. Of the original total of 60 patients with 292 initial carious lesions, after probiotic therapy and dental restoration, 78 total restorations were placed in the subject group over the following three years. Approximately half of these restorations were required in teeth that had initially presented with smaller lesions and had been placed in a "watch" category. Two of the patients that developed further carious lesions had been randomly assigned to the probiotic PerioBalance, while the other two caries active patients were assigned EvoraKids probiotic. Of the original group of caries active patients, 24 did not present with any further carious involvement. Another 25 could be categorized as caries static, as the restorations required were substantially less than before probiotic therapy had been begun. The F-ratio, which in this case equals 51.3313, is a ratio of the between-group estimate to the within-group estimate. Since the P-value of the F-test is less than 0.05, there is a statistically significant difference between the means of the 4 variables at the 95.0% confidence level. Conclusion: The tested probiotic supplements had a statistically significant effect on the caries experience of the enrolled subjects.

Donor lymphocyte infusion and methotrexate for immune recovery after T-cell depleted haploidentical transplantation.

CD34+ cell selection minimizes graft-versus-host disease (GVHD) after haploidentical donor stem cell transplant but is associated with slow immune recovery and infections. We report a Phase I/II study of prophylactic donor lymphocyte infusion (DLI) followed by methotrexate (MTX) GVHD prophylaxis after CD34-selected haploidentical donor transplant. A prophylactic DLI was given between day +30 and +42. Rituximab was given with DLI for the last 10 patients. The goal of the study was to determine a DLI dose that would result in a CD4+ cell count > 100/µL at Day +120 in ≥ 66% of patients with ≤ 33% grade II-III, ≤ 17% grade III, and no grade IV acute GVHD by Day +180. Thirty-five patients with malignant (n = 25) or nonmalignant disease (n = 10) were treated after CD34-selected haploidentical donor peripheral blood stem cell transplant. The DLI dose of 5 × 104 /kg met the CD4/GVHD goal with 67% of patients having CD4+ cells > 100/µL and 11% grade II-IV acute GVHD. The cumulative incidence of chronic GVHD was 16%. Fatal viral and fungal infections occurred in 11%. The 2 year estimated overall survival was 69% and the relapse rate was 14% for patients in remission at transplant. There was no effect of NK alloreactivity on relapse. Nine of ten patients at the target DLI dose cohort of 5 × 104 /kg are alive with median follow-up of 18 mos (range 6-29). Delayed prophylactic DLI and MTX was associated with promising outcomes at the target DLI dose. This trial was registered at clinicaltrials.gov, # NCT01027702.

Primate pulpal healing after exposure and TheraCal application.

AIM: The purpose of this in vivo study was to compare the effectiveness of a new light cured resin based dicalcium/tricalcium silicate pulp capping material (TheraCalLC, Bisco), pure Portland cement, resin based calcium hydroxide or glass ionomer in the healing of bacterially contaminated primate pulps. STUDY DESIGN: The experiment required four primates each having 12 teeth prepared with buccal penetrations into the pulpal tissues with an exposure of approximately 1.0 mm. The exposed pulps of the primate teeth were covered with cotton pellets soaked in a bacterial mixture consisting of microorganisms normally found in human pulpal abscesses. After removal of the pellet, hemostasis was obtained and the pulp capping agents applied. The light cured resin based pulp capping material (TheraCal LC) was applied to the pulpal tissue of twelve teeth with a needle tip syringe and light cured for 15 seconds. Pure Portland cement mixed with a 2% Chlorhexidine solution was placed on the exposed pulpal tissues of another twelve teeth. Twelve additional teeth had a base of GIC applied (Triage, Fuji VII GC America) and another twelve had a pulp cap with VLC DYCAL (Dentsply), a light cured calcium hydroxide resin based material. The pulp capping bases were then covered with a RMGI (Fuji II LC GC America). The tissue samples were collected at 4 weeks. The samples were deminerilized, sectioned, stained and histologically graded. RESULTS: There were no statistically significant differences between the groups in regard to pulpal inflammation (H = 0.679, P = 1.00). However, both the Portland cement and light cured TheraCal LC groups had significantly more frequent hard tissue bridge formation at 28 days than the GIC and VLC Dycal groups (H = 11.989, P = 0.009). The measured thickness of the hard tissue bridges with the pure Portland and light cured TheraCal LC groups were statistically greater than that of the other two groups (H = 15.849, P = 0.002). In addition, the occurrence of pulpal necrosis was greater with the GIC group than the others. Four premolars, one each treated according to the protocols were analyzed with a microCT machine. The premolar treated with the light cured TheraCal LC demonstrated a complete hard tissue bridge. The premolar treated with the GIC did not show a complete hard tissue bridge while the premolar treated with VLC Dycal had an incomplete bridge. The pure Portland with Chlorhexidine mixture created extensive hard tissue bridging. CONCLUSION: TheraCal LC applied to primate pulps created dentin bridges and mild inflammation acceptable for pulp capping.

Cancer Cell Line Inhibition by Osmotic Pump-administered Xylitol in a Syngeneic Mouse Model.

Background: This study aimed to evaluate the effects of continuous administration of xylitol (a commonly used dental prebiotic) via a subcutaneous osmotic minipump in a B16F10 syngeneic mouse model. Methods: The B16F10 syngeneic model consisted of 6-8-week-old C57BL/6 male mice subcutaneously injected with five × 105 B16F10 cells suspended in 100 µl PBS in the right flank. The mice were randomly assigned to two groups: Group 1 was the treatment group with 10% xylitol-loaded pumps (n=10), while Group 2 was the control group with saline-loaded pumps (n=10). Alzet minipumps were implanted subcutaneously in the left flank of B16F10-injected mice once more than 50% of all mice developed palpable tumors. After pump implantation surgery, the mice were monitored daily and weighed 2-3x/week. Tumor sizes were measured with calipers 2-3x/week, and all mice were euthanized when their tumors became too large (20 mm on any axis or 2,000 mm3). The excised tumors were weighed and cut in half, with one half sent for histology and the other for metabolomic analysis. Results: The xylitol-treated group survived substantially longer than the control group. The tumor size was reduced by approximately 35% by volume. Histological sections of xylitol treat mice suggested reduced infiltration and angiogenesis, which is consistent with previous studies. The metabolomic analysis demonstrates that xylitol reduces the tumor production of histamine, NADP+, ATP, and glutathione from the tumor, thereby improving the host immune response with ROS reactive oxygen species. Conclusions: The results of this study suggest that xylitol has potential as an adjunct to oncological treatment and is being further investigated in comparison to monoclonal antibody therapy (Opdualag).

Effect of treatment with phosphate, casein phosphopeptide and fluoride on the remineralization: in vitro study.

This study aimed to evaluate in vitro the effect protocols and anticaries agents containing casein amorphous calcium fluoride phosphopeptide-phosphate (CPP-ACPF, MI Paste Plus), sodium trimetaphosphate (TMP) and fluoride (F), in remineralization of caries lesions. Bovine enamel blocks with initial caries lesions were divided into groups (n = 12): 1) Toothpaste without F-TMP-MI Plus (Placebo); 2) Toothpaste 1100 ppm F (1100F), 3) 1100F + MI Paste Plus (1100F-MI Paste Plus), 4) Toothpaste with 1100F + Neutral gel with 4,500 ppm F + 5%TMP (1100F + Gel TMP) and 5) Toothpaste with 1100F + Neutral gel with 9,000 ppm F (1100F + Gel F). For the 4 and 5 groups the gel was applied only once for 1 minute, initially to the study. For the 3 group, after treatment with 1100F, MI Paste Plus was applied 2x/day for 3 minute. After pH cycling, the percentage of surface hardness recovery (%SHR); integrated loss of subsurface hardness (ΔKHN); profile and depth of the subsuperficial lesion (PLM); concentrations of F, calcium (Ca) and phosphorus (P) in enamel was determined. The data were analyzed by ANOVA (1-criterion) and Student-Newman-Keuls test (p < 0.001). Treatment with 1100F alone led to ~ 28% higher remineralization when compared to treatment with 1100F associated with MI Paste Plus (p < 0.001). The 1100F and 1100F + Gel F groups showed similar values for %SHR (p = 0.150). 1100F + Gel TMP treatment also remineralized the enamel surface by ~ 30% and 20% when compared to the 1100F + Gel F and 1100F groups (p < 0.001). The lower lesion depth (ΔKHN) was observed for the 1100F + Gel TMP group (p < 0.001), where it was 54% and 44% lower in comparison to the 1100F and 1100F + Gel F groups (p < 0.001). Polarized light microscopy photomicrographs showed subsurface lesions in all groups, but these lesions were present to a lower extent in the 1100F + Gel TMP group (p < 0.001). Treatment with 1100F + Gel TMP promoted an increase in the concentration of Ca in the enamel by ~ 57% and ~ 26% when compared to the 1100F and 1100F + MI Paste Plus groups (p < 0.001), respectively. There were no significant differences between the 1100F, 1100F + MI Paste Plus and 1100F + Gel F groups (p > 0.001). Similar values of P in the enamel were observed in the 1100F, 1100F + MI Paste Plus and 1100F + Gel F groups (p > 0.001), except for the 1100F + Gel TMP group, which presented a high concentration (p < 0.001). We conclude that the 1100F+TMP gel treatment/protocol led to a significant increased remineralization when compared to the other treatments/protocols and may be a promising strategy for patients with early caries lesions.

Law enforcement and the long gun: do we need a new face in the fight?

BACKGROUND: The threat of rifles in the hands of criminals is now well recognized within law enforcement. Current emergency response systems are not equipped to operate in this combat-like environment. Growing statistics indicate that of the peace officers that were killed in the line of duty in the United States nearly half died by gunfire evidence. OBJECTIVE: As Emergency Medical Services ("EMS") training and standards evolve, the lessons learned from the Tactical Combat Casualty Care doctrine should be incorporated to improve the safety and outcomes of injured law enforcement officers. Statistics show that deaths by gunfire have the highest average percentage of all officer deaths. Although new weapons, armor, and tactics are continually evolving to meet the challenge of officer safety, in the past decade, little has changed in how our EMS system responds to a critically wounded officer. DISCUSSION: Combat data from the US military leads us to believe that to save a wounded officer, emergency care must start immediately, regardless of the ongoing gun battle. CONCLUSION: It is time for the emergency medical system to evolve to meet the critical needs of today's law enforcement environment.

Biochemical and microbiological characteristics of in situ biofilm formed on materials containing fluoride or amorphous calcium phosphate.

PURPOSE: To evaluate the biochemical and microbiological characteristics of in situ biofilm formed on materials that release fluoride (F-) or calcium (Ca++) and phosphate (Pi). METHODS: This study comprised an in situ and in vitro experiment, utilizing three materials [Auralay XF and Fuji IX GP, containing fluoride, and Aegis containing amorphous calcium phosphate (ACP)] and bovine dental enamel slabs. For the in situ: 10 volunteers wore palatal devices, each containing four material specimens or enamel slabs that were treated with 20% sucrose solution. The biofilm had pH measurements on Day 7 and the composition was analyzed on Day 8 by assessing the following: F-, Ca++, Pi and insoluble extracellular polysaccharides (EPS) concentrations, and then identification of the microbiota. For the in vitro: materials/enamel were subjected to a 7-day pH-cycling regimen to determine F, Ca++ and Pi release. RESULTS: The biofilm formed on F(-)-releasing materials was richer in F, Ca++ and Pi and had lower mutans streptococci counts than enamel biofilm. The biofilm on the ACP-containing material exhibited similar Ca++ and Pi concentrations to biofilm on F(-)-releasing materials. The materials showed buffering action compared with enamel. Biochemical and microbiological characteristics showed a less cariogenic biofilm on materials containing fluoride or amorphous calcium phosphate.

Effectiveness of CRT at measuring the salivary level of bacteria in caries prone children with probiotic therapy.

AIM: This IRB approved clinical trial was to determine the effect of "over the counter" probiotic supplements on the Caries Risk Test- CRT- (Ivoclar) results of the oral microflora in high caries risk children. STUDY DESIGN: Sixty subjects 6 to 12 years old with a caries risk assessment (CAMBRA) of moderate to high (caries prone) were evaluated by an analysis of the difference in the salivary levels of pathogenic bacteria (mutans streptococci and Lactobacilli). The subjects were randomly selected by randomizing software and assigned to two diferent Groups. Group A used PerioBalance (Lactobacilli reuteri-CFU of 200 million) lozenges for 28 days. Group B used the EvoraKids (Streptococcus uberis KJ2, Streptococcus oralis KJ3, Streptococcus rattus JH1 45, > or = 100 million) probiotics chewable tablets for 30 days. Salivary samples were collected then incubated for 48 hours for colony counting and ranking. Follow up testing with the CRT was performed after 60 days at a follow up visit. RESULTS: There was a statistically significant diference in the CRT results between the pre and post use of the probiotics. PerioBalance; SM results t= -6. 78, p< .0001, Lactobacilli results t= -5.762, p< .0001, EvoraKids SMresults t= -7.33, p< .0001, Lactobacilli results t= -2.952, p= .0068. CONCLUSIONS: The CRT values obtained with caries prone children may be significantly affected by probiotic use. Based on this study's results the following conclusions can be made: Both EvoraKids and PerioBalance affected the CRT results by significantly decreasing the number of S. mutans and lactobacilli present in the salivary samples.

In vitro effect of low-fluoride toothpaste supplemented with sodium trimetaphosphate, xylitol, and erythritol on enamel demineralization.

OBJECTIVE: Regular use of toothpaste with fluoride (F) concentrations of ≥ 1000 ppm has been shown to contribute to reducing caries increment. However, when used by children during the period of dental development, it can lead to dental fluorosis. In this study, we aimed to evaluate the in vitro effect of a toothpaste formulation with reduced fluoride (F) concentration (200 ppm) supplemented with sodium trimetaphosphate (TMP: 0.2%), Xylitol (X:16%), and Erythritol (E: 4%) on dental enamel demineralization. METHODOLOGY: Bovine enamel blocks were selected according to initial surface hardness (SHi) and then divided into seven experimental toothpaste groups (n=12). These groups included 1) no F-TMP-X-E (Placebo); 2) 16% Xylitol and 4% Erythritol (X-E); 3) 16% Xylitol, 4% Erythritol and 0.2%TMP (X-E-TMP); 4) 200 ppm F (no X-E-TMP: (200F)); 5) 200 ppm F and 0.2% TMP (200F-TMP); 200 ppm F, 16% Xylitol, 4% Erythritol, and 0.2% TMP (200F-X-E-TMP); and 7) 1,100 ppm F (1100F). Blocks were individually treated 2×/day with slurries of toothpastes and subjected to a pH cycling regimen for five days (DES: 6 hours and RE: 18 hours). Then, the percentage of surface hardness loss (%SH), integrated loss of subsurface hardness (ΔKHN), fluoride (F), calcium (Ca), and phosphorus (P) in enamel were determined. The data were analyzed by ANOVA (1-criterion) and the Student-Newman-Keuls test (p<0.001). RESULTS: We found that the 200F-X-E-TMP treatment reduced %SH by 43% compared to the 1100F treatments (p<0.001). The ΔKHN was ~ 65% higher with 200F-X-E-TMP compared to 1100F (p<0.001). The highest concentration of F in enamel was observed on the 1100F treatment (p<0.001). The 200F-X-E-TMP treatment promote higher increase of Ca and P concentration in the enamel (p<0.001). CONCLUSION: The association of 200F-X-E-TMP led to a significant increase of the protective effect on enamel demineralization compared to the 1100F toothpaste.

Focal adhesion kinase is required for KSHV vGPCR signaling.

Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiologic agent of Kaposi's sarcoma, an angiogenic and inflammatory endothelial cell (EC) tumor that is common in areas of high KSHV prevalence. KSHV encodes a pro-angiogenic viral chemokine receptor (vGPCR) that promotes EC growth in vitro and KS-like tumors in mouse models. vGPCR is therefore considered a viral oncogene that plays a crucial role in the pathobiology of KS. In this study, we show that focal adhesion kinase (FAK) becomes activated upon vGPCR expression in primary ECs and that FAK is required for vGPCR-mediated activation of ERK1/2, NFκB, AP-1, and vGPCR-induced migration and inhibition of anoikis. FAK is crucial to cell motility and tumor invasiveness and is a potential therapeutic target in various malignancies. Our data show that via vGPCR, KSHV has evolved a way to constitutively activate FAK signaling.

The Kaposi's sarcoma-associated herpesvirus G protein-coupled receptor contains an immunoreceptor tyrosine-based inhibitory motif that activates Shp2.

The Kaposi's sarcoma-associated herpesvirus (KSHV) G protein-coupled receptor (vGPCR) is a constitutively active, highly angiogenic homologue of the interleukin-8 (IL-8) receptors that signals in part via the cytoplasmic protein tyrosine phosphatase Shp2. We show that vGPCR contains a bona fide immunoreceptor tyrosine-based inhibitory motif (ITIM) that binds and constitutively activates Shp2.

Effect of poly(ADP-ribose) polymerase 1 on integration of the adeno-associated viral vector genome.

BACKGROUND: Adeno-associated virus type 2 (AAV) has the ability to target integration of its DNA into a specific locus of the human genome. Site-specific AAV integration is mediated by viral Rep proteins, although the role of cellular factors involved in this process is largely unknown. Recent studies provide evidence showing that cellular DNA repair proteins are involved in targeted integration of AAV, although their specific roles are not well defined. METHODS: In the present study, we investigated the interaction between Rep and proteins of the back-up nonhomologous end-joining pathway (B-NHEJ). We then analyzed the effect of one of these proteins, poly(ADP-ribose) polymerase 1 (PARP1) on AAV integration. RESULTS: We show that AAV Rep interacts with B-NHEJ members DNA ligase III and PARP1 but does not associate with the scaffolding factor XRCC1. Moreover, PARP1 and Rep bind directly and not via DNA-protein interactions. We also found that Rep increases the enzymatic activity of PARP1 potentially through the endonuclease activity of Rep. Finally, we demonstrate that both chemical inhibition of PARP1 and PARP1 depletion using small hairpin RNA enhance integration of the AAV genome in HeLa cells. CONCLUSIONS: The findings of the present study indicate that manipulation of PARP1 activity could be used as a tool for developing new, effective AAV-based therapies for the treatment of genetic diseases and cancer.

Non-integrating lentiviral vectors for specific killing of Epstein-Barr virus nuclear antigen 1-positive B cell lymphoma cells.

BACKGROUND: Epstein-Barr virus (EBV) causes a range of life-threatening B-lymphocyte malignancies but, despite the use of various strategies, treatment remains problematic. METHODS: In the present study, we developed a non-integrating lentiviral vector (NILV) that mediates specific killing of EBV nuclear antigen 1 (EBNA1)-expressing cells with minimal toxicity to EBNA1-negative cells. The EBV family of repeats (FR) was cloned intok the NILV genome upstream of various transgenes. RESULTS: The presence of the FR in the NILV genome induced transcriptional up-regulation and prolonged the expression of a transgene specifically in EBNA1-positive B cells. Transgene expression from an FR-containing NILV was also prolonged in EBV-transformed cells compared to an FR-negative NILV. We found that the delivery of an FR-containing NILV encoding herpes simplex virus 1 thymidine kinase (TK) lead to the killing of more than 99% of EBNA1-positive B cells with minimal toxicity to EBNA1-negative cells in the presence of gancyclovir. EBNA1-positive cells were not killed by an FR-negative vector containing the TK gene. An FR-TK-containing NILV also specifically killed EBNA1-containing cells in a mixed population of EBNA1-positive and EBNA1-negative cells, thus confirming that NILV-FR-TK-mediated killing is specific for EBNA1-expressing cells. CONCLUSIONS: Transgene expression from our NILVs is both EBNA1-specific and dependent upon the presence of the FR. The results obtained in the present study indicate that NILVs have potential use in the treatment of EBV-associated B cell malignancies.

Effect of fluoride, casein phosphopeptide-amorphous calcium phosphate and sodium trimetaphosphate combination treatment on the remineralization of caries lesions: An in vitro study.

OBJECTIVE: To evaluate the effects of combination of treatments with fluoridated toothpastes supplemented with sodium trimetaphosphate (TMP) and casein phosphopeptide-amorphous calcium phosphate (MI Paste Plus®), on the remineralization of dental enamel. DESIGN: Enamel blocks with artificial caries were randomly allocated into six groups (n = 12), according to the toothpastes: 1) without F-TMP-MI Paste Plus® (Placebo); 2) 1100 ppm F (1100 F), 3) MI Paste Plus®, 4) 1100 F + MI Paste Plus® (1100 F-MI Paste Plus®), 5) 1100 F + 3% TMP (1100 F-TMP) and 6) 1100 F-TMP + MI Paste Plus® (1100 F-TMP-MI Paste Plus®). Blocks were treated 2×/day with slurries of toothpastes (1 min). Furthermore, groups 4 and 6 received the application of MI Paste Plus® for 3 min. After pH cycling, the percentage of surface hardness recovery (%SHR); integrated loss of subsurface hardness (ΔKHN); profile analysis and lesion depth subsurface through polarized light microscopy (PLM), confocal laser scanning microscopy (LSCM), scanning electron microscopy (SEM), fluoride (F), calcium (Ca), phosphorus (P) concentrations in the enamel were determined. The data were analyzed by ANOVA (1-criterion) and Student-Newman-Keuls test (p < 0.001). RESULTS: 1100 F-TMP-MI Paste Plus® group showed the best results of %SHR, ΔKHN and PLM (p < 0.001). F concentration was similar between the 1100 F, 1100 F-MI Paste Plus®, and 1100 F-TMP-MI Paste Plus® groups (p > 0.001). 1100 F-TMP-MI Paste Plus® group showed the highest concentration of Ca and P in the enamel (p < 0.001). CONCLUSION: The association of 1100 F-TMP and MI Paste Plus® led to a significant increase in the remineralization of initial carious lesions.

In situ evaluation of 200 ppm fluoride toothpaste content trimetaphosphate, xylitol and erythritol on enamel demineralization and dental biofilm.

OBJECTIVE: To evaluate the effect of low-fluoride (F-) toothpaste and sodium trimetaphosphate (TMP) associated with xylitol and erythritol (XE) on enamel demineralization and biofilm composition. METHODS: This crossover double-blind in situ study consisted of five phases (seven days each), in which 14 volunteers wore oral appliances containing four enamel bovine blocks. The cariogenic challenge was performed by exposure to a 30% sucrose solution (6x/day). The toothpaste treatments (3x/day) were as follows: placebo (no F-/TMP/XE); 200 ppm F- (NaF) (200F); 1,100 ppm F- (1100F); 16% Xylitol and 4% Erythritol (XE); and 200 ppm F-, 0.2% TMP, 16% xylitol, and 4% erythritol (200F-TMP-XE). Percentage of surface hardness loss (%SH) and integrated loss of subsurface hardness (ΔKHN), and calcium (Ca2+), phosphate (PO43-), and F- on enamel and biofilm were determined; as well as insoluble extracellular polysaccharide (EPS). RESULTS: XE and 1100F groups showed no significant difference for %SH and ΔKHN values (p = 0.220 and p = 0.886), and the 200F-TMP-XE group had the lowest mineral loss (p < 0.001). Ca2+ and PO43- in the enamel showed the highest values (p < 0.001) for the 200F-TMP-XE group. Higher values of F- in the enamel and biofilm were observed for the 1100F group (p < 0.001). There was no difference for Ca2+ (p = 1.00) and EPS (p =0.918) values between XE and 200-TMP-XE groups in the biofilm, but their values were higher and lower than the 1100F (p = 0.002 and p = 0.029), respectively. CONCLUSIONS: 200F-TMP-XE promoted a greater protective effect against enamel demineralization and significantly affected the composition of biofilm formed in situ compared to 1100F toothpaste. CLINICAL SIGNIFICANCE: Low-F- toothpaste containing TMP and polyols can be considered an effective and safe measure to improve the oral health of individuals, especially patients with high caries activity.

KSHV LANA inhibits TGF-beta signaling through epigenetic silencing of the TGF-beta type II receptor.

Signaling through the transforming growth factor-beta (TGF-beta) pathway results in growth inhibition and induction of apoptosis in various cell types. We show that this pathway is blocked in Kaposi sarcoma herpesvirus (KSHV)-infected primary effusion lymphoma through down-regulation of the TGF-beta type II receptor (TbetaRII) by epigenetic mechanisms. Our data also suggest that KSHV infection may result in lower expression of TbetaRII in Kaposi sarcoma and multicentric Castleman disease. KSHV-encoded LANA associates with the promoter of TbetaRII and leads to its methylation and to the deacetylation of proximal histones. Reestablishment of signaling through this pathway reduces viability of these cells, inferring that KSHV-mediated blockage of TGF-beta signaling plays a role in the establishment and progression of KSHV-associated neoplasia. These data suggest a mechanism whereby KSHV evades both the antiproliferative effects of TGF-beta signaling by silencing TbetaRII gene expression and immune recognition by suppressing TGF-beta-responsive immune cells through the elevated secretion of TGF-beta1.

In situ evaluation of the remineralizing capacity of pit and fissure sealants containing amorphous calcium phosphate and/or fluoride.

OBJECTIVE: This in situ study evaluated the remineralizing potential of pit and fissure sealants containing amorphous calcium phosphate (ACP) and/or fluoride in artificially induced carious lesions on smooth enamel surfaces. MATERIAL AND METHODS: Ten volunteers who wore acrylic palatal devices were enrolled in this 5-day double-blind study and assigned to one of the following five groups: (I) demineralized enamel slab+Fluroshield (sealant with fluoride); (II) demineralized enamel slab+Aegis (sealant with ACP); (III) demineralized enamel slab+experimental sealant with fluoride (ESF); (IV) demineralized enamel slab+experimental sealant with fluoride/ACP (ACP-F); and (V) demineralized enamel slab (control). After the experimental period, the percentage of surface microhardness recovery (%SMHR) and the integrated loss of subsurface hardness (DeltaKHN) were evaluated. The concentrations of fluoride, calcium and phosphorus in enamel were also determined. RESULTS: The sealants containing ACP and/or fluoride presented a higher remineralizing capacity (%SMHR and DeltaKHN) than that of the control group. Aegis provided either more efficient or similar remineralization than the other sealants. The association between ACP and fluoride did not show a greater efficacy in the remineralization. F, Ca and P concentrations in enamel varied according to the group. CONCLUSION: The pit and fissure sealants containing ACP were able to promote remineralization of artificially induced carious lesions on smooth enamel surfaces.