Influence of BMI percentile on craniofacial morphology and development in children and adolescents.

BACKGROUND: The prevalence of childhood and adolescent obesity is increasing worldwide. Reports suggest that elevated body mass index (BMI) is associated with larger craniofacial dimensions and advanced dental and skeletal development. Such an association is important for timing orthodontic treatment relative to pubertal growth and dental eruption. MATERIALS AND METHODS: To evaluate associations between BMI, craniofacial morphology, dental age, and cervical vertebrae maturation staging (CVMS), 400 participants were consecutively selected (8-15 years, n = 200 overweight and obese BMI >85%, 200 normal weight) from the University of North Carolina database. Records were analysed for cephalometric measures, Demirjian index values, and CVMS. Bivariate statistics and linear regression analysis evaluated whether CVMS, dental age, and cephalometric dimensions varied with BMI. RESULTS: Overweight/obese children and adolescents had a proportionally larger bimaxillary prognathic skeletal pattern compared to those of normal weight. These cephalometric measurements [articulare-gnathion (Ar-Gn), condylion-anterior nasal spine (Co-ANS), sella-gonion (S-Go), nasion-menton (N-Me), anterior nasal spine-menton (ANS-Me), sella-nasion-A point (SNA), sella-nasion-B point (SNB), and sella-nasion-pogonion (SNPg)] were significantly different [statistically (P < 0.05) and clinically (>2 mm or >2 degrees)] between the two study groups, with a linear relationship between BMI percentile and craniofacial dimension. The overweight/obese BMI group had a mean dental age 1.4 years advanced relative to the normal weight group (P < 0.05), with an advancement of nearly one CVM stage between the ages of 12 and 14 (P < 0.05). LIMITATIONS: The study is retrospective. CONCLUSIONS: Obese/overweight children and adolescents have proportionally larger antero-posterior and vertical dimensions and are more likely to experience advanced dental and skeletal maturation. Obese/overweight subjects may enter their growth spurt at a younger age and have earlier eruption of teeth, affecting treatment timing. BMI percentile should be a consideration for orthodontic treatment in growing patients.

Post-prophylaxis gingivitis prevention with two-step stannous fluoride dentifrice plus whitening gel sequence or chlorhexidine gluconate mouthrinse.

PURPOSE: To assess use of a two-step dentifrice/gel sequence versus chlorhexidine gluconate mouthrinse on gingivitis prevention after dental prophylaxis. METHODS: A 12-week, randomized controlled trial was conducted to compare the effectiveness and safety of a two-step dentifrice/gel sequence to a positive control in healthy adults with established gingivitis. After informed consent, gingivitis and stain levels were assessed by clinical examination. Eligible subjects received a dental prophylaxis and were randomly assigned to twice daily unsupervised use of either (1) two-step oral hygiene sequence:0.454% stannous fluoride dentifrice followed by 3.0% hydrogen peroxide whitening gel for the test group; or (2) 0.12% chlorhexidine gluconate oral rinse and 0.76% sodium monofluorophosphate dentifrice for the control group. Clinical measurements of gingivitis bleeding sites and tooth stain area/intensity were collected after 4, 8 and 12 weeks use, while safety was assessed via clinical examination and oral status interview of the subjects. RESULTS: A total of 44 subjects were enrolled and 35 completed the 12-week study. At baseline, bleeding sites ranged from 10-33. After prophylaxis and assigned treatment, both groups exhibited significant (P≤ 0.0001) reductions in bleeding sites. Responses were directionally better in the two-step sequence at all post-baseline timepoints, with groups differing significantly (P < 0.05) at Week 8. Tooth stain measurements demonstrated that the two-step dentifrice/gel sequence did not contribute to any significant (P> 0.13) stain accumulation. In contrast, stain accumulation was evident (P< 0.003) in the chlorhexidine group beginning at the Week 4 visit. Adverse events were more common in the positive control, and contributed to early termination. CLINICAL SIGNIFICANCE: Twice daily use of a two-step stannous fluoride dentifrice and peroxide whitening gel sequence after prophylaxis provided comparable or superior gingivitis benefits to chlorhexidine gluconate rinse without the concomitant side effect of staining.

A new HDV mouse model identifies mitochondrial antiviral signaling protein (MAVS) as a key player in IFN-ß induction.

BACKGROUND & AIMS: Studying hepatitis delta virus (HDV) and developing new treatments is hampered by the limited availability of small animal models. Herein, a description of a robust mouse model of HDV infection that mimics several important characteristics of the human disease is presented. METHODS: HDV and hepatitis B virus (HBV) replication competent genomes were delivered to the mouse liver using adeno-associated viruses (AAV; AAV-HDV and AAV-HBV). Viral load, antigen expression and genomes were quantified at different time points after AAV injection. Furthermore, liver pathology, genome editing, and the activation of the innate immune response were evaluated. RESULTS: AAV-HDV infection initiated HDV replication in mouse hepatocytes. Genome editing was confirmed by the presence of small and large HDV antigens and sequencing. Viral replication was detected for 45days, even after the AAV-HDV vector had almost disappeared. In the presence of HBV, HDV infectious particles were detected in serum. Furthermore, as observed in patients, co-infection was associated with the reduction of HBV antigen expression and the onset of liver damage that included the alteration of genes involved in the development of liver pathologies. HDV replication induced a sustained type I interferon response, which was significantly reduced in immunodeficient mice and almost absent in mitochondrial antiviral signaling protein (MAVS)-deficient mice. CONCLUSION: The animal model described here reproduces important characteristics of human HDV infection and provides a valuable tool for characterizing the viral infection and for developing new treatments. Furthermore, MAVS was identified as a main player in HDV detection and adaptive immunity was found to be involved in the amplification of the innate immune response. Lay summary: Co-infection with hepatitis B and D virus (HBV and HDV, respectively) often causes a more severe disease condition than HBV alone. Gaining more insight into HDV and developing new treatments is hampered by limited availability of adequate immune competent small animal models and new ones are needed. Here, a mouse model of HDV infection is described, which mimics several important characteristics of the human disease, such as the initiation and maintenance of replication in murine hepatocytes, genome editing and, in the presence of HBV, generation of infectious particles. Lastly, the involvement of an adaptive immunity and the intracellular signaling molecule MAVS in mounting a strong and lasting innate response was shown. Thus, our model serves as a useful tool for the investigation of HDV biology and new treatments.

Effect of Two-minute Application of 35% Sodium Ascorbate on Composite Bond Strength following Bleaching.

AIM: The aim of this study is to assess the effect of 35% sodium ascorbate on microtensile bond strength of dentin immediately after bleaching with 35% hydrogen peroxide. MATERIALS AND METHODS: A total of 25 sound human 3rd molars were collected. Teeth were randomly divided into five groups for different treatments: Group I [bleaching + immediate bonding (i.e., restoration)], group II (bleaching + delayed bonding), group III (bleaching + sodium ascorbate + immediate bonding), group IV (bleaching + sodium ascorbate + delayed bonding), and group V (bonding only). After bleaching, but before bonding, groups II and IV were stored for 1 week in deionized water at 37°C. All samples were bonded using OptiBoned FL (Kerr) and Filtek Supreme (3M/ESPE). Teeth were sectioned into 1 × 1 mm 2 bars, and microtensile bond strength was tested with a universal testing machine (Instron 8841) at a cross-head speed of 0.5 mm/minute. RESULTS: Microtensile bond strength differed significantly across the five groups, with a significant reduction in microtensile bond strength observed for samples in group I relative to samples in any of the other treatment groups (p < 0.05). CONCLUSION: The application of a high concentration of sodium ascorbate for a shorter time reversed the negative effect of 35% hydrogen peroxide bleaching on composite bonding strength to dentin. CLINICAL SIGNIFICANCE: The negative effects of bleaching on composite bonding can be neutralized by the application of the reversing agent sodium ascorbate thus, increasing the efficiency of clinic chair time. This is clinically relevant for those patients requiring restorative treatment immediately after in-office bleaching.

Inhibition of SARS-CoV-2 infection by Porphyromonas gingivalis and the oral microbiome.

The COVID-19 pandemic persists despite the availability of vaccines, and it is therefore crucial to develop new therapeutic and preventive approaches. In this study, we investigated the potential role of the oral microbiome in SARS-CoV-2 infection. Using an in vitro SARS-CoV-2 pseudovirus infection assay, we found a potent inhibitory effect exerted by Porphyromonas gingivalis on SARS-CoV-2 infection mediated by known P. gingivalis compounds such as phosphoglycerol dihydroceramide (PGDHC) and gingipains as well as by unknown bacterial factors. We found that the gingipain-mediated inhibition of infection is likely due to cytotoxicity, while PGDHC inhibited virus infection by an unknown mechanism. Unidentified factors present in P. gingivalis supernatant inhibited SARS-CoV-2 likely via the fusion step of the virus life cycle. We addressed the role of other oral bacteria and found certain periodontal pathogens capable of inhibiting SARS-CoV-2 pseudovirus infection by inducing cytotoxicity on target cells. In the human oral cavity, we observed the modulatory activity of oral microbial communities varied among individuals in that some saliva-based cultures were capable of inhibiting while others were enhancing infection. These findings contribute to our understanding of the complex relationship between the oral microbiome and viral infections, offering potential avenues for innovative therapeutic strategies in combating COVID-19.

Comparative study of weekly and three-weekly neoadjuvant carboplatin in triple-negative breast cancer: retrospective analysis in real-world settings.

PURPOSE: Tri-weekly carboplatin is an established neoadjuvant treatment for triple-negative breast cancer, enhancing pathological complete response (pCR) and overall survival. This study explores if weekly carboplatin provides lower toxicity and comparable pCR rates. METHODS/PATIENTS: A retrospective multicenter study (January 2021 to March 2023) compares outcomes of weekly and tri-weekly carboplatin. RESULTS: Among 104 participants, 60% received weekly and 40% tri-weekly treatments. Weekly administration had fewer discontinuations (56.5 vs. 70.7%, p = 0.154). Both schedules exhibited similar overall toxicity (p = 0.087), with slightly higher grade 3-4 toxicity in the tri-weekly group (56.1 vs. 48.4%, p = 0.126). Hematological toxicity was comparable, but the weekly group experienced more diarrhea (p = 0.432) and asthenia (p = 0.012). Weekly treatment correlated with more frequent breast-conserving surgeries (p = 0.004). pCR rates were 50% with weekly and 61% with tri-weekly regimens (p = 0.186). CONCLUSIONS: Weekly carboplatin exhibited comparable toxicity, a trend toward fewer interruptions, and similar pCR rates. Prospective studies are essential for validating these findings.

Sophisticated viral quasispecies with a genotype-related pattern of mutations in the hepatitis B X gene of HBeAg-ve chronically infected patients.

Patients with HBeAg-negative chronic infection (CI) have not been extensively studied because of low viremia. The HBx protein, encoded by HBX, has a key role in viral replication. Here, we analyzed the viral quasispecies at the 5' end of HBX in CI patients and compared it with that of patients in other clinical stages. Fifty-eight HBeAg-negative patients were included: 16 CI, 19 chronic hepatitis B, 16 hepatocellular carcinoma and 6 liver cirrhosis. Quasispecies complexity and conservation were determined in the region between nucleotides 1255 and 1611. Amino acid changes detected were tested in vitro. CI patients showed higher complexity in terms of mutation frequency and nucleotide diversity and higher quasispecies conservation (p < 0.05). A genotype D-specific pattern of mutations (A12S/P33S/P46S/T36D-G) was identified in CI (median frequency, 81.7%), which determined a reduction in HBV DNA release of up to 1.5 log in vitro. CI patients showed a more complex and conserved viral quasispecies than the other groups. The genotype-specific pattern of mutations could partially explain the low viremia observed in these patients.

Conservation and variability of hepatitis B core at different chronic hepatitis stages.

BACKGROUND: Since it is currently not possible to eradicate hepatitis B virus (HBV) infection with existing treatments, research continues to uncover new therapeutic strategies. HBV core protein, encoded by the HBV core gene (HBC), intervenes in both structural and functional processes, and is a key protein in the HBV life cycle. For this reason, both the protein and the gene could be valuable targets for new therapeutic and diagnostic strategies. Moreover, alterations in the protein sequence could serve as potential markers of disease progression. AIM: To detect, by next-generation sequencing, HBC hyper-conserved regions that could potentially be prognostic factors and targets for new therapies. METHODS: Thirty-eight of 45 patients with chronic HBV initially selected were included and grouped according to liver disease stage [chronic hepatitis B infection without liver damage (CHB, n = 16), liver cirrhosis (LC, n = 5), and hepatocellular carcinoma (HCC, n = 17)]. HBV DNA was extracted from patients' plasma. A region between nucleotide (nt) 1863 and 2483, which includes HBC, was amplified and analyzed by next-generation sequencing (Illumina MiSeq platform). Sequences were genotyped by distance-based discriminant analysis. General and intergroup nt and amino acid (aa) conservation was determined by sliding window analysis. The presence of nt insertion and deletions and/or aa substitutions in the different groups was determined by aligning the sequences with genotype-specific consensus sequences. RESULTS: Three nt (nt 1900-1929, 2249-2284, 2364-2398) and 2 aa (aa 117-120, 159-167) hyper-conserved regions were shared by all the clinical groups. All groups showed a similar pattern of conservation, except for five nt regions (nt 1946-1992, 2060-2095, 2145-2175, 2230-2250, 2270-2293) and one aa region (aa 140-160), where CHB and LC, respectively, were less conserved (P < 0.05). Some group-specific conserved regions were also observed at both nt (2306-2334 in CHB and 1935-1976 and 2402-2435 in LC) and aa (between aa 98-103 in CHB and 28-30 and 51-54 in LC) levels. No differences in insertion and deletions frequencies were observed. An aa substitution (P79Q) was observed in the HCC group with a median (interquartile range) frequency of 15.82 (0-78.88) vs 0 (0-0) in the other groups (P < 0.05 vs CHB group). CONCLUSION: The differentially conserved HBC and HBV core protein regions and the P79Q substitution could be involved in disease progression. The hyper-conserved regions detected could be targets for future therapeutic and diagnostic strategies.

Effect of a desensitizing paste containing 8% arginine and calcium carbonate on the surface roughness of dental materials and human dental enamel.

PURPOSE: To evaluate the effect of an 8% arginine-calcium carbonate fluoride-free desensitizing paste on the surface roughness of resin composite, porcelain, amalgam, gold, and human dental enamel both prior to and following simulated toothbrushing. METHODS: A resin composite (Filtek Supreme), a commercial porcelain (IPS Empress), an amalgam (Dispersalloy), gold (JIF-PF) and human dental enamel were used, as well as commercial finishing and polishing instruments. Eight two-sided samples were fabricated for each group. The composite and amalgam samples were stored at 100% relative humidity and 37 degrees C for 48 hours prior to measuring the surface roughness and completing the subsequent finishing and polishing procedures. Enamel blocks were cut from human lesion-free teeth and embedded in acrylic. The blocks were then polished flat with high polishing pastes. For gold and porcelain, the same size was used and the materials processed by a professional dental laboratory. Following storage, each surface was polished using the Super-Snap (Shofu) system. The amalgam was polished with conventional polishing techniques. Roughness (Ra and Ry) was evaluated with both a 3D non-contact profilometer and a stylus profilometer. With the two-sided samples only one side was polished with the desensitizing paste and the other side was left unpolished without paste. The 8% arginine-calcium carbonate desensitizing paste was applied to a surface for 15 seconds using a single disposable prophy cup. Each polished surface was measured by the profilometers and three roughness values per surface were recorded as the "initial prophy" surface. Following initial surface analysis, each side of every sample was treated with a simulated toothbrushing technique using a toothbrushing device (V-8). A 50:50 (w/w) slurry of toothpaste (Colgate Cavity Protection) and deionized water was used. Each surface was brushed 10,000 times. Then, the samples were rinsed with tap water and stored in 100% humidity until roughness values were obtained using the profilometers as previously described ("toothbrush surface"). After analyzing the brushed surfaces, the samples were returned to their original treatment group ("desensitizing paste"). Each surface was re-polished with the desensitizing paste as previously stated. Those surfaces (referred to as "recall paste") were measured as previously described for final surface roughness. Data was analyzed using repeated measures two-factor ANOVA with Tukey HSD pairwise comparison as appropriate (alpha=0.05). Two additional samples were made of each material in order to measure step-heights. Tape was placed on the surface of each sample to separate the treatment side and the non-treated side. The tape was removed before each profilometry reading. RESULTS: The desensitizing paste containing 8% arginine and calcium carbonate did not have a significant effect on the surface roughness of the substrates tested. Although the 3D non-contact profilometry images showed slight roughness after toothbrushing followed by the use of the desensitizing paste, these changes were not statistically significant (P>0.05).

Sealant adaptation and penetration into occlusal fissures.

PURPOSE: To evaluate the adaptation and penetration into occlusal fissures of two different types of fissure sealants. METHODS: Extracted third molars (n=10) with evident occlusal fissures were cleaned with a pumice/water slurry and randomly divided into two groups and sealed following the manufacturers' directions as follows: Group 1--Embrace fissure sealant (Pulpdent). Surfaces were cleaned and dried, then etched for 15 seconds. Excess water was removed leaving the surface slightly moist. Sealant was applied from cusp to cusp without covering marginal ridges and light cured for 20 seconds using a halogen light at 500 mW/cm2. Group 2--ClinPro (3M Espe). Surfaces were cleaned and dried then etched for 15 seconds. The etched surface was rinsed and thoroughly dried. Dried surfaces appeared frosty white. Sealant was placed making sure not to go beyond etched area, and light cured for 20 seconds. Teeth were thermocycled (500x) and sectioned with an Isomet in a mesio-distal direction (4 slices per tooth). The sections were examined under the SEM. The marginal adaptation of the sealants was evaluated under the SEM using the following criteria: 1 = Smooth adaptation. Sealant flows with enamel. No ledges; 2 = Sealant is not well adapted. Ledge may be present. The penetration ability of the sealants was evaluated under the SEM using the following criteria: 1 = Sealant penetrated 1/3 the total length of the fissure; 2 = Sealant penetrated 1/2 the total length of the fissure; 3 = Sealant penetrated the total length of the fissure. The results were statistically analyzed using a t-test. RESULTS: Embrace showed consistently more intimate marginal adaptation than ClinPro in fissures of the same approximate width and depth (P < 0.05).

Characterization of hepatitis B virus X gene quasispecies complexity in mono-infection and hepatitis delta virus superinfection.

BACKGROUND: Hepatitis delta virus (HDV) seems to strongly suppress hepatitis B virus (HBV) replication, although little is known about the mechanism of this interaction. Both these viruses show a dynamic distribution of mutants, resulting in viral quasispecies. Next-generation sequencing is a viable approach for analyzing the composition of these mutant spectra. As the regulatory hepatitis B X protein (HBx) is essential for HBV replication, determination of HBV X gene (HBX) quasispecies complexity in HBV/HDV infection compared to HBV mono-infection may provide information on the interactions between these two viruses. AIM: To compare HBV quasispecies complexity in the HBX 5' region between chronic hepatitis delta (CHD) and chronic HBV mono-infected patients. METHODS: Twenty-four untreated patients were included: 7/24 (29.2%) with HBeAg-negative chronic HBV infection (CI, previously termed inactive carriers), 8/24 (33.3%) with HBeAg-negative chronic hepatitis B (CHB) and 9/24 (37.5%) with CHD. A serum sample from each patient was first tested for HBV DNA levels. The HBX 5' region [nucleotides (nt) 1255-1611] was then PCR-amplified for subsequent next-generation sequencing (MiSeq, Illumina, United States). HBV quasispecies complexity in the region analyzed was evaluated using incidence-based indices (number of haplotypes and number of mutations), abundance-based indices (Hill numbers of order 1 and 2), and functional indices (mutation frequency and nucleotide diversity). We also evaluated the pattern of nucleotide changes to investigate which of them could be the cause of the quasispecies complexity. RESULTS: CHB patients showed higher median HBV-DNA levels [5.4 logIU/mL, interquartile range (IQR) 3.5-7.9] than CHD (3.4 logIU/mL, IQR 3-7.6) (P = n.s.) or CI (3.2 logIU/mL, IQR 2.3-3.5) (P < 0.01) patients. The incidence and abundance indices indicated that HBV quasispecies complexity was significantly greater in CI than CHB. A similar trend was observed in CHD patients, although only Hill numbers of order 2 showed statistically significant differences (CHB 2.81, IQR 1.11-4.57 vs CHD 8.87, 6.56-11.18, P = 0.038). There were no significant differences in the functional indices, but CI and CHD patients also showed a trend towards greater complexity than CHB. No differences were found for any HBV quasispecies complexity indices between CHD and CI patients. G-to-A and C-to-T nucleotide changes, characteristic of APOBEC3G, were higher in CHD and CI than in CHB in genotype A haplotypes, but not in genotype D. The proportion of nt G-to-A vs A-to-G changes and C-to-T vs T-to-C changes in genotype A and D haplotypes in CHD patients showed no significant differences. In CHB and CI the results of these comparisons were dependent on HBV genotype. CONCLUSION: The lower-replication CHD and CI groups show a trend to higher quasispecies complexity than the higher-replication CHB group. The mechanisms associated with this greater complexity require elucidation.

Clinical Profile of Autism Spectrum Disorder in a Pediatric Population from Northern Mexico.

Autism spectrum disorder (ASD) is a neurodevelopmental condition classified based on needs of support, in order to address impairments in the areas of social communication and restricted and repetitive behavior. The aim of this work is to describe the main clinical features of the ASD severity levels in a group of Mexican pediatric patients. The results show firstly that this condition was more frequent in males than females. Secondly, an inverse relationship was found between the intellectual coefficient and the level of severity of the disorder. Thirdly, deficits in social reciprocity and communication were more evident in Level 3, than in Levels 1 and 2, while the difference was less evident in restricted and repetitive patterns of behavior.

The effects of high concentration tooth whitening bleaches on microleakage of Class V composite restorations.

OBJECTIVE: To explore the effects of high-concentration hydrogen peroxide bleaching agents on the microleakage of composite restorations. METHODS: In 60 extracted human molars, Class V restorations were prepared with Scotchbond 1/Filtek Z250 composite. Teeth were randomly divided into four groups: (1) no bleaching; (2) bleaching with 14% hydrogen peroxide gel from Crest Whitestrips; (3) bleaching with 20% carbamide peroxide gel from Opalescence PF 20; and (4) bleaching with 38% hydrogen peroxide gel Opalescence Xtra Boost. Bleaching procedures were carried out at 37 degrees C for 21 days/42 hours (2); seven days/42 hours (3); one day/45 minutes (4). Varnish was applied on the apical portion of the teeth only, excluding the restoration, prior to immersion in a 0.1% rhodamin-B-isothiocyanate solution for 24 hours at 37 degrees C. After rinsing, specimens were embedded in methacrylate blocks, and sectioned with a water-cooled microtome with three restoration cuts positioned centrally parallel to the long axis of the tooth. Microleakage was evaluated at the occlusal margins of the Class V restorations using a stereo microscope, separate for dentin and enamel margins. RESULTS: Over 90% of enamel margins exhibited no microleakage following cycling. Bleaching agents had almost no effect on numerical averages. Eighty-eight percent of the dentin margins were free of microleakage for the non-treated control group. Bleaching treatments collectively had slight numerical reductions to around 80%. The statistical evaluation (Kruskal-Wallis-test) showed no significant difference in microleakage between groups for enamel or dentin. CONCLUSION: Bleaching with the materials tested had no influence on microleakage of Filtek Z250 composite bonded with Scotchbond 1.

Quantitative characterization of hepatitis delta virus genome edition by next-generation sequencing.

AIM: To determine the capacity of next-generation sequencing (NGS) for quantifying edited and unedited HDV populations, and to confirm if edition is a general phenomenon taking place along the entire HDV region analyzed, as we previously reported (Homs M et al. PLoS One 2016, 11, e0158557). METHODS: Four serum samples from 4 patients with chronic HDV/HBV infection were included in the study. The region selected for analysis covered 360 nucleotides (nt), positions 910-1270 of the HDV genome, which included the HDAg ORF editing site (nt 1014 within codon 196). Quantification of edited and unedited genomes was performed by molecular cloning and Sanger sequencing and by NGS. To evaluate the reliability of the NGS values obtained, we combined a clone with an edited codon and one with an unedited codon in known percentages in a series of artificial mixtures, which were then analyzed by NGS. In addition, we determined the nt changes occurring over the complete amplified region after excluding the editing codon (196) to evaluate edition along it. RESULTS: In total, 11,208 quality-filtered sequences were obtained in the 4 samples. The 95% confidence intervals for the proportions of unedited populations by molecular cloning and NGS were overlapping, and those of cloning were wider, indicating that they are comparable and that NGS is more precise than cloning. Unedited genomes predominated over edited ones in all 4 samples analyzed by NGS and in 3 of the 4 samples analyzed by molecular cloning. In total, 83,276 quality-filtered sequences were obtained from the artificial mixtures. Percentages of the two viral populations detected by NGS in these mixtures were comparable to the expected percentages. Evaluation of edition along the HDV coding region showed that transitions were more frequent than transversions, accounting for 63.09% and 36.91%, respectively. Interestingly, among the 4 possible transition-type changes, G:A and A:G accounted for 73.86% of the total. CONCLUSION: Next-generation sequencing proved useful to quantify edited and unedited HDV genomes, and provided relevant information on the HDV quasispecies.

A Comparison of Oral Hygiene Products and Professional Care: A six-week randomized clinical trial.

Purpose: To investigate the anti-gingivitis efficacy of a novel oral hygiene routine consisting of a two-step stannous fluoride dentifrice and hydrogen peroxide whitening gel system, an interactive oscillating-rotating electric toothbrush, and expanded polytetrafluoroethylene floss.Methods: A total of 52 participants (n=52;mean age 35.8±11.23 years) were enrolled in the study and randomized 1:1 to the experimental hygiene group or control (dental prophylaxis followed by use of standard sodium fluoride dentifrice and a manual toothbrush). Participants were instructed to brush twice daily; those in the experimental group were instructed to floss once daily. Oral examinations were conducted at Baseline, Week 2, Week 4, and Week 6.Results: Both groups experienced significant declines in the mean number of bleeding sites from Baseline at all time points, evident as early as Week 2. Bleeding sites continued to decline throughout the trial in the experimental group, whereas they showed an increasing trend between Weeks 2 and 6 in the control group. The experimental group had 55% fewer bleeding sites at Week 2, 85% fewer bleeding sites at Week 4, and 98% fewer bleeding sites at Week 6 (p<0.0001 for all) as compared to the control group. At Week 6, 84% of participants in the experimental group had no bleeding, while all participants in the control group had bleeding.Conclusion: The experimental oral hygiene group showed significantly greater reductions in gingival bleeding than the control oral hygiene group, with benefits seen as early as Week 2 and increasing over the six-week study.

Detection of hyper-conserved regions in hepatitis B virus X gene potentially useful for gene therapy.

AIM: To detect hyper-conserved regions in the hepatitis B virus (HBV) X gene (HBX) 5' region that could be candidates for gene therapy. METHODS: The study included 27 chronic hepatitis B treatment-naive patients in various clinical stages (from chronic infection to cirrhosis and hepatocellular carcinoma, both HBeAg-negative and HBeAg-positive), and infected with HBV genotypes A-F and H. In a serum sample from each patient with viremia > 3.5 log IU/mL, the HBX 5' end region [nucleotide (nt) 1255-1611] was PCR-amplified and submitted to next-generation sequencing (NGS). We assessed genotype variants by phylogenetic analysis, and evaluated conservation of this region by calculating the information content of each nucleotide position in a multiple alignment of all unique sequences (haplotypes) obtained by NGS. Conservation at the HBx protein amino acid (aa) level was also analyzed. RESULTS: NGS yielded 1333069 sequences from the 27 samples, with a median of 4578 sequences/sample (2487-9279, IQR 2817). In 14/27 patients (51.8%), phylogenetic analysis of viral nucleotide haplotypes showed a complex mixture of genotypic variants. Analysis of the information content in the haplotype multiple alignments detected 2 hyper-conserved nucleotide regions, one in the HBX upstream non-coding region (nt 1255-1286) and the other in the 5' end coding region (nt 1519-1603). This last region coded for a conserved amino acid region (aa 63-76) that partially overlaps a Kunitz-like domain. CONCLUSION: Two hyper-conserved regions detected in the HBX 5' end may be of value for targeted gene therapy, regardless of the patients' clinical stage or HBV genotype.

How is the biocompatibilty of dental biomaterials evaluated?

All biomaterials used in dentistry must be evaluated for biocompatibility using screening assays to protect patient health and safety. The purpose of this review is to explain the international biocompatibility guidelines, and to explain the structure of a test program. The test program requires the structured assessment of materials into four phases; general toxicity, local tissue irritation, pre-clinical, and clinical evaluation. Different types of screening assays are available, and it is important to understand the advantages and limitations of the various types of assays that are available, so that they can be selected for appropriateness and interpreted accurately. New scientific advances in terms of the chemical properties of dental materials, tissue engineering, stem cell, genetic transfer, biomaterial, and growth factor therapies are under development. These new therapies create improved opportunities to restore and regenerate oral tissues, but they can also present new hazards to patients. Prior to their clinical use, these new technologies must be proven to be safe, and not hazardous to human health. A structured biocompatibility assessment and advice on the selection of assays are outlined to evaluate these new therapies.

Evidence of an Exponential Decay Pattern of the Hepatitis Delta Virus Evolution Rate and Fluctuations in Quasispecies Complexity in Long-Term Studies of Chronic Delta Infection.

Chronic HDV infection can cause a severe form of viral hepatitis for which there is no specific treatment. Characterization of the hepatitis B or C viral quasispecies has provided insight into treatment failure and disease recurrence following liver transplantation, has proven useful to understand hepatitis B e antigen seroconversion, and has helped to predict whether hepatitis C infection will resolve or become chronic. It is likely that characterization of the hepatitis delta virus (HDV) quasispecies will ultimately have similar value for the management of this infection. This study sought to determine the RNA evolution rates in serum of chronic hepatitis delta (CHD) treatment-naïve patients, using next-generation sequencing methods. The region selected for study encompassed nucleotide positions 910 to 1270 of the genome and included the amber/W codon. Amber/W is a substrate of the editing process by the ADAR1 host enzyme and is essential for encoding the 2 delta antigens (HDAg). The amber codon encodes the small (unedited) HDAg form and the W codon the large (edited) HDAg form. The evolution rate was analyzed taking into account the time elapsed between samples, the percentage of unedited and edited genomes, and the complexity of the viral population. The longitudinal studies included 29 sequential samples from CHD patients followed up for a mean of 11.5 years. In total, 121,116 sequences were analyzed. The HDV evolution rate ranged from 9.5x10-3 to 1.2x10-3 substitutions/site/year and showed a negative correlation with the time elapsed between samples (p<0.05). An accumulation of transition-type changes was found to be responsible for higher evolution rates. The percentages of unedited and edited genomes and the quasispecies complexity showed no relationships with the evolution rate, but the fluctuations in the percentages of genomes and in complexity suggest continuous adaptation of HDV to the host conditions.