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The aim of this study was to present a new technique for plastic carrier removal in retreatment situations. The gutta-percha technique that uses plastic carrier obturators is one of the most commonly used; however, removing this core is tedious and time-consuming. Two clinical cases were performed by an expert in the field of endodontics, to expose this technique, in which a conical ultrasonic tip with an active point and a Hëdstrom hand file allowed the removal of the plastic carrier quickly and effectively. The use of magnification is indispensable for a predictable success with this method SI

Aims: To discuss the terminology to define and classify actinic cheilitis (AC) and to build a consensus on the diagnostic and therapeutic approaches to AC. Methods: Two-round Delphi study using a questionnaire including 34 closed sentences (9 on terminology and taxonomy, 5 on potential for malignant transformation, 12 on diagnostic aspects, 8 on treatment) and 8 open questions. Experts’ agreement was rated using a Likert scale (1–7). Results: A consensus was reached on 24 out 34 statements (73.5%) and on 5 out of 8 (62.5%) close-ended questions. The response rate was identical in both rounds (attrition of 0%). AC is the term with the highest agreement (median of 7 (strongly agree; IQR: 6–7)) and the lowest dispersion (VC = 21.33). ‘Potentially malignant disorder’ was the preferred classification group for AC (median of 7) and 85.6% of participants showing some level of agreement (CV < 50). Experts (66.75%) consider AC a clinical term (median: 7; IQR: 4–7) and believe definitive diagnosis can be made clinically (median: 6; IQR: 5–7), particularly by inspection and palpation (median: 5; IQR: 4–6). Histopathological confirmation is mandatory for the management of AC (median: 5; IQR: 2.5–7), even for homogeneous lesions (median: 5; IQR: 3.5–6). Consensus was reached on all treatment statements (VC < 50). Conclusions: AC is a potentially malignant disorder with a significant lack of agreement on diagnostic criteria, procedures, biopsy indications and the importance of techniques to assist in biopsy. A consensus was reached on nomenclature and management of this disorder SI

Objective: To undertake a meta-analysis of the association of Oral Lichen Planus (OLP) with diabetes, two diseases with an important impact on public health and the economy, but the evidence of which about their association is inconsistent. Methods: Relevant studies were localized by searching MEDLINE, EMBASE, Conference Proceedings, and other databases from inception to October 2020, without restrictions. The reference lists of included studies and of related reviews were also inspected. Global pooled odds ratios were calculated, and predefined subgroup analyses were performed. The heterogeneity between studies and publication bias was assessed and sensitivity analysis was carried out. Results: Thirty-two studies were included in the meta-analysis. Pooled ORs showed a moderate association between diabetes and OLP [OR: 1.87 (95%CI: 1.57, 2.34)]. The association is limited to studies carried out on adults only [OR: 2.12 (95%CI: 1.75, 2.57)] and is observed in all study designs. Globally, the heterogeneity was low to moderate. Studies carried out in European populations show a stronger association of diabetes and OLP than Asiatic studies [OR: 2.49 (95%CI: 1.87, 3.32) and 1.60 (95%CI: 1.25, 2.03), respectively]. Conclusions: Diabetes and OLP are moderately associated. Systematic diagnosis of diabetes in OLP patients could prove useful SI

Background: The aim was to investigate the clinical significance of nestin immunohistochemical expression in head and neck area lesions and to study its role in patient survival and recurrence. Methods: 39 (44.3%) nasosinus, 37 (42%) major salivary gland (6 submandibular and 31 parotid) and 12 (13.6%) oral cavity lesions of paraffin-embedded samples were retrospectively included. Results: The expression was categorized into grades, negative for 55 (62.5%) cases, grade 1 in 10 cases (11.4%), grade 2 in 12 cases (13.6%), and grade 3 in 11 cases (12.5%), 100% of pleomorphic adenomas were positive for nestin with grade 3 intensity, 100% of polyps and inverted papillomas were negative (p &lt, 0.001). The lowest estimate of disease-free-survival (DFS) was for grade 1 expression, with 50 months, confidence interval (CI): 95% 13.3–23.9 months and the highest for grade 3 expression, 167.9 months (CI: 95% 32.1–105 months, Log-Rank = 14.846, p = 0.002). ROC (receiver operating characteristic) curves revealed that the positivity for nestin (+/−) in relation to malignancy, presented a sensitivity of 50.98%, a specificity of 81.08%, with an area under the curve of 0.667 (p = 0.009). Conclusions: Nestin could be a useful marker to detect the presence of stem cells in head and neck tumors that have a role in tumor initiation and progression.

This study aimed to investigate the impact of COVID-19 on the experiences of special care dentistry providers worldwide. An online survey was administered from 10 to 31 July 2020. Age, sex, years of professional activity, COVID-19 status, geographical area of origin and length of lockdown period were recorded for all participating dentists. The relationships between these variables and the changes in clinical activity, the treated patients’ COVID-19 status and the implementation of protective measures in the dental clinic were analyzed. A total of 436 (70.6% women) dentists from 59 countries responded to the survey. Clinical activity was reduced or stopped for 79.1% of respondents. The most common change was to limit treatment to urgent care only (53.7%). Treatment under general anesthesia or deep sedation was discontinued (51.0%) or reduced (35.8%) for the majority of respondents. Male dentists were more likely to maintain their clinical activity than female dentists (p &lt, 0.001), and respondents from North America were more likely to do so than participants from other geographical regions (p &lt, 0.001). Dentists from Latin America and the Caribbean were more likely to report treatment of confirmed cases of COVID-19 than those from Europe (p &lt, 0.001). The implementation of protective measures in the dental office was determined by the survey participant’s sex, intensity of clinical activity and geographical area of origin. To conclude, the provision of special care dentistry was considerably reduced in response to the pandemic. Service maintenance was mainly related to the geographical area in which the surveyed dentists worked, further exacerbating pre-existing inequalities.

Uveitis is a vision inflammatory disorder with a high prevalence in developing countries. Currently, marketed treatments remain limited and reformulation is usually performed to obtain a tacrolimus eye drop as a therapeutic alternative in corticosteroid-refractory eye disease. The aim of this work was to develop a mucoadhesive, non-toxic and stable topical ophthalmic formulation that can be safely prepared in hospital pharmacy departments. Four different ophthalmic formulations were prepared based on the tacrolimus/hydroxypropyl-&beta, cyclodextrin (HP&beta, CD) inclusion complexes&rsquo, formation. Phase solubility diagrams, Nuclear Magnetic Resonance (NMR) and molecular modeling studies showed the formation of 1:1 and 1:2 tacrolimus/HP&beta, CD inclusion complexes, being possible to obtain a 0.02% (w/v) tacrolimus concentration by using 40% (w/v) HP&beta, CD aqueous solutions. Formulations also showed good ophthalmic properties in terms of pH, osmolality and safety. Stability studies proved these formulations to be stable for at least 3 months in refrigeration. Ex vivo bioadhesion and in vivo ocular permanence showed good mucoadhesive properties with higher ocular permanence compared to the reference pharmacy compounding used in clinical settings (t1/2 of 86.2 min for the eyedrop elaborated with 40% (w/v) HP&beta, CD and Liquifilm&reg, versus 46.3 min for the reference formulation). Thus, these novel eye drops present high potential as a safe alternative for uveitis treatment, as well as a versatile composition to include new drugs intended for topical ophthalmic administration.

The objective of this study was to conduct a systematic review and meta-analysis on the efficacy of sentinel lymph node biopsy (SLNB) in T1/T2-N0 oral squamous cell carcinoma (OSCC). A systematic review of the literature on SLNB until March 2019 was conducted. The review was organized according to the PRISMA protocol, considering the following PICO (population, intervention, comparison, outcome) question: What is the sensitivity of sentinel lymph node biopsy in OSCC? ‘P’ was patients with head and neck squamous cell carcinoma T1/2-N0; ‘I’ was SLNB; ‘C’ was neck treated with elective neck dissection and haematoxylin–eosin histopathology; ‘O’ was sensitivity and specificity. A meta-analysis and meta-regression were performed on the selected studies. The sensitivity of SLNB was up to 88% (95% confidence interval (CI) 72–96%) and specificity was up to 99% (95% CI 96–100%). The area under the summary receiver operating characteristic curve was 0.99 (95% CI 0.98–1.00). In the four studies where immunohistochemistry was performed, both the sensitivity and specificity were higher than in the studies without immunohistochemistry: 93% (95% CI 88–97%) and 98% (95% CI 96–100%), respectively. In conclusion, SLNB is an effective technique for treating patients with some types of stage T1/2-N0 OSCC. Some parameters such as immunohistochemistry could determine the level of diagnostic accuracy SI

Aim: To study clinical, radiographic, and microbiological outcomes after non-surgical therapy of peri-implantitis with or without adjunctive systemic metronidazole. Materials and Methods: A randomized placebo-controlled clinical trial was carried out in 32 subjects (62 implants) diagnosed with peri-implantitis. Implants received a mechanical non-surgical debridement session and systemic metronidazole or placebo. Clinical, radiographic, and microbiological outcomes were evaluated at baseline, 3, 6, and 12 months. Results: After 12 months, the test treatment resulted in significantly greater PPD reduction (2.53 vs. 1.02 mm) and CAL gain (2.14 vs. 0.53 mm) (p value

Purpose The shape of the mandible has been analyzed in a variety of fields, whether to diagnose conditions like osteoporosis or osteomyelitis, in forensics, to estimate biological information such as age, gender, and race or in orthognathic surgery. Although the methods employed produce encouraging results, most rely on the dry bone analyses or complex imaging techniques that, ultimately, hamper sample collection and, as a consequence, the development of large-scale studies. Thus, we proposed an objective, repeatable, and fully automatic approach to provide a quantitative description of the mandible in orthopantomographies (OPGs). Methods We proposed the use of a deep convolutional neural network (CNN) to localize a set of landmarks of the mandible contour automatically from OPGs. Furthermore, we detailed four different descriptors for the mandible shape to be used for a variety of purposes. This includes a set of linear distances and angles calculated from eight anatomical landmarks of the mandible, the centroid size, the shape variations from the mean shape, and a group of shape parameters extracted with a point distribution model. Results The fully automatic digitization of the mandible contour was very accurate, with a mean point to the curve error of 0.21 mm and a standard deviation comparable to that of a trained expert. The combination of the CNN and the four shape descriptors was validated in the well-known problems of forensic sex and age estimation, obtaining 87.8% of accuracy and a mean absolute error of 1.57 years, respectively. Conclusion The methodology proposed, including the shape model, can be valuable in any field that requires a quantitative description of the mandible shape and a visual representation of its changes such as clinical practice, surgery management, dental research, or legal medicine.

Introduction: The etiologies of most chronic temporomandibular joint disorders are unknown. However, an association between habitual chewing on a particular side and chronic temporomandibular joint disorders has been reported. The aim of this study was to investigate the differences between sides (affected vs unaffected) of biodynamic factors (including lateral dental guidance determined by dental anatomy) or condylar path angles (determined by temporomandibular joint morphology) and chewing function (physiological alternate chewing vs single habitual chewing side). The study scope was to investigate possible etiological factors to improve the understanding of temporomandibular joint disorders. The null hypothesis was that no difference would be found between sides that are or are not affected by chronic temporomandibular joint disorders in chewing function or in levels of dental or temporomandibular joint remodeling. Methods: This cross-sectional, double-blind study involved 24 adults with substantial, chronic, unilateral symptoms diagnosed as temporomandibular joint disorders. Chewing function, temporomandibular joint remodeling (using axiography) and dental anatomy (lateral guidance angles using kinesiography) were assessed. Results: Habitual chewing on one particular side was observed in 17 of 24 participants; significantly more (n = 15) chewed on the affected side than on the unaffected side (P = 0.002 in a two-tailed Fisher’s exact test; risk estimate = 4.5; 95% CI 1.326–15.277). The condylar path (CP) angle was steeper on the affected side than on the unaffected side (mean (standard deviation) = 50.52° (9.98°) versus 45.50° (7.98°); P = 0.002 in a two-tailed t-test). The lateral guidance (LG) angles were flatter on the affected side in all 24 participants. Conclusion: Our results suggest that habitual chewing on one side may be associated with increasing condylar path, with flattening lateral guidance angles, and also with chronic temporomandibular joint disorder on the habitual chewing side This study was supported by the Carlos III Institute of Health (Ministry of Science and Innovation of the Government of Spain and the European Development Fund, ‘Una manera de hacer Europa’) (grant no. PI11/02507). The design, management, analysis, and reporting of the study are entirely independent of the Carlos III Institute of Health SI

DNA hypermethylation is an important epigenetic mechanism for gene expression inactivation in head and neck cancer (HNC). Saliva has emerged as a novel liquid biopsy representing a potential source of biomarkers. We performed a comprehensive meta-analysis to evaluate the overall diagnostic accuracy of salivary DNA methylation for detecting HNC. PubMed EMBASE, Web of Science, LILACS, and the Cochrane Library were searched. Study quality was assessed by the Quality Assessment for Studies of Diagnostic Accuracy-2, and sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (dOR), and their corresponding 95% confidence intervals (CIs) were calculated using a bivariate random-effect meta-analysis model. Meta-regression and subgroup analyses were performed to assess heterogeneity. Eighty-four study units from 18 articles with 8368 subjects were included. The pooled sensitivity and specificity of salivary DNA methylation were 0.39 and 0.87, respectively, while PLR and NLR were 3.68 and 0.63, respectively. The overall area under the curve (AUC) was 0.81 and the dOR was 8.34. The combination of methylated genes showed higher diagnostic accuracy (AUC, 0.92 and dOR, 36.97) than individual gene analysis (AUC, 0.77 and dOR, 6.02). These findings provide evidence regarding the potential clinical application of salivary DNA methylation for HNC diagnosis This work was co-funded by the Instituto de Salud Carlos III (ISCIII) (PI20/01449) and the European Regional Development Fund (FEDER). A.D.-L. is funded by a “Juan Rodés” contract from ISCIII (JR17/00016). L.M.-R. is funded by a “Miguel Servet” contract from ISCIII (CP20/00129) SI

9 pages, 6 figures.-- Under a Creative Commons license, Numerous therapeutic strategies have been developed for osteoarthritis (OA) management, including intra-articular (IA) injections. The ideal IA formulation should control cartilage degradation and restore synovial fluid viscosity. To this end, we propose to combine thermo-sensitive polymers (poloxamers) with hyaluronic acid (HA) to develop suitable beta-lapachone (βLap) loaded IA formulations. The development of IA formulations with these components entails several difficulties: low βLap solubility, unknown βLap therapeutic dose and the bonded commitment of easy administration and viscosupplementation. An optimized formulation was designed using artificial intelligence tools based on the experimental results of a wide variety of hydrogels and its therapeutic capacity was evaluated on an ex vivo OA model. The formulation presented excellent rheological properties and significantly decreased the secretion of degradative (MMP13) and pro-inflammatory (CXCL8) molecules. Therefore, the developed formulation is a promising candidate for OA treatment restoring the synovial fluid rheological properties while decreasing inflammation and cartilage degradation, Financial support was received from Interreg V-A POCTEP (0245_IBEROS_1_E) EU (FEDER). Research groups are also supported by Xunta de Galicia (Competitive Reference Groups, ED431C 2020/17-FEDER) and (Grupos de Potencial Crecimiento, IN607B 2018/19-FEDER)

The present study used 16S rRNA gene amplicon sequencing to assess the impact on salivary microbiome of different grades of dental and periodontal disease and the combination of both (hereinafter referred to as oral disease), in terms of bacterial diversity, co-occurrence network patterns and predictive models. Our scale of overall oral health was used to produce a convenience sample of 81 patients from 270 who were initially recruited. Saliva samples were collected from each participant. Sequencing was performed in Illumina MiSeq with 2 × 300 bp reads, while the raw reads were processed according to the Mothur pipeline. The statistical analysis of the 16S rDNA sequencing data at the species level was conducted using the phyloseq, DESeq2, Microbiome, SpiecEasi, igraph, MixOmics packages. The simultaneous presence of dental and periodontal pathology has a potentiating effect on the richness and diversity of the salivary microbiota. The structure of the bacterial community in oral health differs from that present in dental, periodontal or oral disease, especially in high grades. Supragingival dental parameters influence the microbiota’s abundance more than subgingival periodontal parameters, with the former making a greater contribution to the impact that oral health has on the salivary microbiome. The possible keystone OTUs are different in the oral health and disease, and even these vary between dental and periodontal disease: half of them belongs to the core microbiome and are independent of the abundance parameters. The salivary microbiome, involving a considerable number of OTUs, shows an excellent discriminatory potential for distinguishing different grades of dental, periodontal or oral disease; considering the number of predictive OTUs, the best model is that which predicts the combined dental and periodontal status This investigation was supported by the Instituto de Salud Carlos III (General Division of Evaluation and Research Promotion, Madrid, Spain) and co-financed by FEDER (“A way of making Europe”) under grant ISCIII/PI17/01722, and the CESPU under grants MVOS2016 and MVOS-PT-IINFACTS-2019 SI

Background: Wolf-Hirschhorn syndrome (WHS) is a rare disease caused by deletion in the distal moiety of the short arm of chromosome 4. The objectives of this study were to report the most representative oral findings of WHS, relate them with other clinical characteristics of the disease, and establish possible phenotype-genotype correlation. Methods: The study was conducted at 6 reference centers distributed throughout Spain during 2018&ndash, 2019. The study group consisted of 31 patients with WHS who underwent a standardized oral examination. Due to behavioral reasons, imaging studies were performed on only 11 of the children 6 years of age or older. All participants had previously undergone a specific medical examination for WHS, during which anatomical, functional, epilepsy-related, and genetic variables were recorded. Results: The most prevalent oral manifestations were delayed tooth eruption (74.1%), bruxism (64.5%), dental agenesis (63.6%), micrognathia (60.0%), oligodontia (45.5%), and downturned corners of the mouth (32.3%). We detected strong correlation between psychomotor delay and oligodontia (p = 0.008, Cram&eacute, r&rsquo, s V coefficient, 0.75). The size of the deletion was correlated in a statistically significant manner with the presence of oligodontia (p = 0.009, point-biserial correlation coefficient, 0.75). Conclusion: Certain oral manifestations prevalent in WHS can form part of the syndrome&rsquo, s phenotypic variability. A number of the characteristics of WHS, such as psychomotor delay and epilepsy, are correlated with oral findings such as oligodontia and bruxism. Although most genotype-phenotype correlations are currently unknown, most of them seem to be associated with larger deletions, suggesting that some oral-facial candidate genes might be outside the critical WHS region, indicating that WHS is a contiguous gene syndrome.

Background and Objectives: Head and Neck Squamous Cell Carcinoma (HNSCC) includes cancers from the oral cavity, larynx, and oropharynx and is the sixth-most common cancer worldwide. MicroRNAs are small non-coding RNAs for which altered expression has been demonstrated in pathological processes, such as cancer. The objective of our study was to evaluate the different expression profile in HNSCC subtypes and the prognostic value that one or several miRNAs may have. Materials and Methods: Data from The Cancer Genome Atlas Program-Head and Neck Squamous Cell Carcinoma (TCGA-HNSCC) patients were collected. Differential expression analysis was conducted by edge R-powered TCGAbiolinks R package specific function. Enrichment analysis was developed with Diana Tool miRPath 3.0. Kaplan-Meier survival estimators were used, followed by log-rank tests to compute significance. Results: A total of 127 miRNAs were identified with differential expression level in HNSCC, 48 of them were site-specific and, surprisingly, only miR-383 showed a similar deregulation in all locations studied (tonsil, mouth, floor of mouth, cheek mucosa, lip, tongue, and base of tongue). The most probable affected pathways based on miRNAs interaction levels were protein processing in endoplasmic reticulum, proteoglycans in cancer (p &lt, 0.01), Hippo signaling pathway (p &lt, 0.01), and Transforming growth factor-beta (TGF-beta) signaling pathway (p &lt, 0.01). The survival analysis highlighted 38 differentially expressed miRNAs as prognostic biomarkers. The miRNAs with a greater association between poor prognosis and altered expression (p &lt, 0.001) were miR-137, miR-125b-2, miR-26c, and miR-1304. Conclusions: In this study we have determined miR-137, miR-125b-2, miR-26c, and miR-1304 as novel powerful prognosis biomarkers. Furthermore, we have depicted the miRNAs expression patterns in tumor patients compared with normal subjects using the TCGA-HNSCC cohort.

Objectives: The COVID-19 (SARS-CoV-2) pandemic is an ongoing public health challenge, also for the dentistry community. The main objective of this paper was to determine the economic and health-care impact of COVID-19 on dentists in the Autonomous Region of Galicia (Spain). Methods: This was a descriptive observational study in which the data was collected by means of a self-administered survey (from 1 April 2020 to 30 April 2020). Results: A total of 400 dentists from Galicia responded to the survey. Only 12.3% of the participants could obtain personal protective equipment (PPE) including FFP2 masks. Of the male respondents, 33.1% suffered losses &gt, &euro, 15,000 compared to 19.4% of female respondents (OR = 3.121, p &lt, 0.001). Economic losses seem to have contributed to the applications for economic help as 29.5% of the respondents who applied for this measure recorded losses in excess of &euro, 15,000 (p = 0.03). Patients complained more about the fact that only emergency care was available during the State of Alarm, in dental surgeries that do not work with insurance companies or franchises. Only 4 professionals tested positive, 50% of whom worked exclusively in private practice and the other 50% who practised in both private and public surgeries. Dentists who practise in the public sector saw more urgent patients per week than those practising in private surgeries (p = 0.013). Conclusions: The COVID-19 pandemic has had economic repercussions in dentistry as only urgent treatment was available during the State of Alarm. These repercussions seem to be higher in male participants, as the majority of the participants have revealed higher economic losses than females. The level of assistance has also been affected, reducing the number of treated patients, although this quantity has been different in private and public surgeries. By presenting these findings we look to highlight the role that dentists play in society in treating dental emergencies in our surgeries, and this must be recognised and addressed by the relevant authorities, who must provide PPEs as a priority to this group as well as providing special economic aid in accordance with the losses incurred by the sector.

Circulating tumor cell (CTC) enumeration has emerged as a powerful biomarker for the assessment of prognosis and the response to treatment in metastatic breast cancer (MBC). Moreover, clinical evidences show that CTC-cluster counts add prognostic information to CTC enumeration, however, their significance is not well understood, and more clinical evidences are needed. We aim to evaluate the prognostic value of longitudinally collected single CTCs and CTC-clusters in a heterogeneous real-world cohort of 54 MBC patients. Blood samples were longitudinally collected at baseline and follow up. CTC and CTC-cluster enumeration was performed using the CellSearch&reg, system. Associations with progression-free survival (PFS) and overall survival (OS) were evaluated using Cox proportional hazards modelling. Elevated CTC counts and CTC-clusters at baseline were significantly associated with a shorter survival time. In joint analysis, patients with high CTC counts and CTC-cluster at baseline were at a higher risk of progression and death, and longitudinal analysis showed that patients with CTC-clusters had significantly shorter survival compared to patients without clusters. Moreover, patients with CTC-cluster of a larger size were at a higher risk of death. A longitudinal analysis of a real-world cohort of MBC patients indicates that CTC-clusters analysis provides additional prognostic value to single CTC enumeration, and that CTC-cluster size correlates with patient outcome.

MET alterations may provide a potential biomarker to evaluate patients who will benefit from treatment with MET inhibitors. Therefore, the purpose of the present study is to investigate the utility of a liquid biopsy-based strategy to assess MET alterations in cancer patients. We analyzed MET amplification in circulating free DNA (cfDNA) from 174 patients with cancer and 49 healthy controls and demonstrated the accuracy of the analysis to detect its alteration in patients. Importantly, a significant correlation between cfDNA concentration and MET copy number (CN) in cancer patients (r = 0.57, p &lt, 10&minus, 10) was determined. Furthermore, we evaluated two approaches to detect the presence of MET on circulating tumor cells (CTCs), using the CellSearch&reg, and Parsortix systems and monitored patients under anti-EGFR treatment (n = 30) combining both cfDNA and CTCs analyses. This follow-up provides evidence for the potential of MET CN assessment when patients develop resistance to anti-EGFR therapy and a significant association between the presence of CTCs MET+ and the Overall Survival (OS) in head and neck cancer patients (P = 0.05, HR = 6.66). In conclusion, we develop specific and noninvasive assays to monitor MET status in cfDNA/CTCs and demonstrate the utility of plasma MET CN determination as a biomarker for monitoring the appearance of resistance to anti-EGFR therapy.

Background: In the dentistry field, the analysis of dental plaque is vital because it is the main etiological factor in the 2 most prevalent oral diseases: caries and periodontitis. In most of the papers published in the dental literature, the quantification of dental plaque is carried out using traditional, non-automated, and time-consuming indices. Therefore, the development of an automated plaque quantification tool would be of great value to clinicians and researchers. Objective: This study aimed to develop a web-based tool called DenTiUS and various clinical indices to evaluate dental plaque levels using image analysis techniques. Methods: The tool was executed as a web-based application to facilitate its use by researchers. Expert users are free to define experiments, including images from either a single patient (to observe an individual plaque growth pattern) or several patients (to perform a group characterization) at a particular moment or over time. A novel approach for detecting visible plaque has been developed as well as a new concept known as nonvisible plaque. This new term implies the classification of the remaining dental area into 3 subregions according to the risk of accumulating plaque in the near future. New metrics have also been created to describe visible and nonvisible plaque levels. Results: The system generates results tables of the quantitative analysis with absolute averages obtained in each image (indices about visible plaque) and relative measurements (indices about visible and nonvisible plaque) relating to the reference moment. The clinical indices that can be calculated are the following: plaque index of an area per intensity (API index, a value between 0 and 100), area growth index (growth rate of plaque per unit of time in hours; percentage area/hour), and area time index (the time in days needed to achieve a plaque area of 100% concerning the initial area at the same moment). Images and graphics can be obtained for a moment from a patient in addition to a full report presenting all the processing data. Dentistry experts evaluated the DenTiUS Plaque software through a usability test, with the best-scoring questions those related to the workflow efficiency, value of the online help, attractiveness of the user interface, and overall satisfaction. Conclusions: The DenTiUS Plaque software allows automatic, reliable, and repeatable quantification of dental plaque levels, providing information about area, intensity, and growth pattern. Dentistry experts recognized that this software is suitable for quantification of dental plaque levels. Consequently, its application in the analysis of plaque evolution patterns associated with different oral conditions, as well as to evaluate the effectiveness of various oral hygiene measures, can represent an improvement in the clinical setting and the methodological quality of research studies This work received financial support from Johnson & Johnson company (Grant 2016‐CE219), Consellería de Cultura, Educación e Ordenación Universitaria (accreditation 2019-2022 ED431G-2019/04, 2017-2020 Potential Growth Group ED431B 2017/029, 2017-2020 Competitive Reference Group ED431C 2017/69, and N. Vila-Blanco support ED481A-2017), and the European Regional Development Fund (ERDF), which acknowledges the CiTIUS-Research Center in Intelligent Technologies of the University of Santiago de Compostela as a Research Center of the Galician University System SI

[Abstract] Carbonic anhydrase IX (CAIX) is a hypoxia-related protein considered as a predictor for oral squamous cell carcinoma (OSCC) biological behaviour. Nevertheless, this prognostic value is still yet to be validated. We aim to quantify prognostic significance of CAIX overexpression in OSCC by meta-analysis. We performed searches in MEDLINE, EMBASE, SCOPUS, WOS, WHO’S databases, CPCI, and OATD from inception to August 2019. Overall survival (OS), disease-free survival (DFS), locoregional control (LC), and disease-specific survival (DSS) were considered as outcomes of interest. Overall 18 studies were included. CAIX overexpression was associated with worse OS (hazard ratio [HR] = 1.45 95% confidence interval [CI] 1.17–1.80) and DFS (HR = 1.98 95% CI 1.18–3.32). To the contrary, it was neither associated with LC (HR = 1.01 95% CI 0.50–2.02) nor with DSS (HR = 1.35 95% CI 0.78–2.33). Heterogeneity was negligible in all analyses except for DSS. Small studies effect was not significant for OS and DFS. This study shows that immunohistochemical CAIX assessment is a useful OSCC prognostic biomarker.

Acyl homoserine lactones (AHLs), the quorum sensing (QS) signals produced by Gram-negative bacteria, are currently considered to play a minor role in the development of oral biofilm since their production by oral pathogens has not been ascertained thus far. However, we report the presence of AHLs in different oral samples and their production by the oral pathogen Porphyromonas gingivalis. The importance of AHLs is further supported by a very high prevalence of AHL-degradation capability, up to 60%, among bacteria isolated from dental plaque and saliva samples. Furthermore, the wide-spectrum AHL-lactonase Aii20J significantly inhibited oral biofilm formation in different in vitro biofilm models and caused important changes in bacterial composition. Besides, the inhibitory effect of Aii20J on a mixed biofilm of 6 oral pathogens was verified using confocal microscopy. Much more research is needed in order to be able to associate specific AHLs with oral pathologies and to individuate the key actors in AHL-mediated QS processes in dental plaque formation. However, these results indicate a higher relevance of the AHLs in the oral cavity than generally accepted thus far and suggest the potential use of inhibitory strategies against these signals for the prevention and treatment of oral diseases This work was supported by the grant “Axudas do Programa de Consolidación e Estructuración de Unidades de Investigación Competitivas (GPC)” from the Consellería de Cultura, Educación e Ordenación Universitaria, Xunta de Galicia (ED431B2017/53) and by the project INTERREG-POCTEP-0227-CODIGOMAIS-1-E. A.M. was supported by a predoctoral fellowship from the Consellería de Cultura, Educación e Ordenación Universitaria, Xunta de Galicia (ED481A-2015/311) SI

Salivary microRNAs (miRNAs) are of high interest as diagnostic biomarkers for non-oral cancer. However, little is known about their value for colorectal cancer (CRC) detection. Our study aims to characterize salivary miRNAs in order to identify non-invasive markers for CRC diagnosis. The screening of 754 miRNAs was performed in saliva samples from 14 CRC and 10 healthy controls. The differential expressed miRNAs were validated by RT-qPCR in 51 CRC, 19 adenomas and 37 healthy controls. Receiver operating characteristic (ROC) curves and logistic regression models were performed to analyze the clinical value of these miRNAs. Twenty-two salivary miRNAs were significantly deregulated in CRC patients vs. healthy individuals (P &lt, 0.05) in the discovery phase. From those, five upregulated miRNAs (miR-186-5p, miR-29a-3p, miR-29c-3p, miR-766-3p, and miR-491-5p) were confirmed to be significantly higher in the CRC vs. healthy group (P &lt, 0.05). This five-miRNA signature showed diagnostic value (72% sensitivity, 66.67% specificity, AUC = 0.754) to detect CRC, which was even higher in combination with carcinoembryonic antigen (CEA) levels. Overall, after the first global characterization of salivary miRNAs in CRC, a five-miRNA panel was identified as a promising tool to diagnose this malignancy, representing a novel approach to detect cancer-associated epigenetic alterations using a non-invasive strategy.

This is the accepted manuscript of the following article: de Coo, A., Quintela, I., Blanco, J., Diz, P., & Carracedo, Á. (2018). Assessment of genotyping tools applied in genetic susceptibility studies of periodontal disease: A systematic review. Archives Of Oral Biology, 92, 38-50. doi: 10.1016/j.archoralbio.2018.04.012. © Elsevier 2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license Objective: A systematic review to evaluate the various genotyping tools and study strategies employed to define genetic susceptibility to periodontitis. Methods: The review was performed in accordance with Preferred Reporting Items for Systematic Review and Meta-Analysis guidelines. The search for publications referring to the genetic bases of periodontal disease was performed on the MEDLINE-PubMed and Cochrane Library databases, on trials registers, and on the web pages of regulatory agencies. Results: We found 2439 potentially eligible articles, of which only 25 satisfied the established inclusion criteria and were processed for data extraction. The review revealed marked heterogeneity between studies, caused in part by the lack of a universally accepted definition for periodontitis phenotypes and by the variety of genotyping tools available. The most commonly used technique was genotyping candidate genes. Conclusion: The few rigorous studies that have been published on genetic susceptibility to periodontitis are subject to severe methodological bias due to their design and the genotyping tools employed. Despite their limitations, candidate gene studies continue to be the predominant methodological approach, rather than genome-wide association studies. Further studies must be designed using a universally accepted, validated diagnostic criterion for periodontitis, analysing multiple genes and polymorphisms in combination with rare variants This review was supported in part by University f Santiago de Composela and by the Spanish Ministry of Education, Culture and Sport by a FPU grant (FPU15/05974) SI

Background: Small nucleolar RNAs (snoRNAs) are small non-coding RNA sequences whose most studied function is ribosome biogenesis. The altered expression of snoRNA is observed in tumoral processes such as breast cancer and multiple myeloma. However, we have not found any references to snoRNAs in oral squamous cell carcinomas (OSCC) in the literature at the time this article was written. Material and methods: We have analyzed snoRNA expression in frozen OSCC tissue samples and have compared them to healthy controls. RNA was extracted from a total of eight OSCC samples and eight control samples, measuring the differential expression of small RNAs with the Affymetrix® miRNA 4.1 Array Plate microarray platform. Results: Results were analyzed using the Transcriptome Analysis Console 3.0 (TAC) software. We obtained a total of 16 deregulated snoRNAs of which one was over expressed and 15 were under expressed. SnoRNAs expression was altered in OSCC and could serve as a diagnostic marker SI

Fluctuating dental asymmetry (FDA) is a tool to measure developmental stability that could be increased in gonosomal aneuploidies. The aim of this study was to quantify FDA in individuals with Down syndrome (DS). The study group comprised 40 individuals with DS, and a control group matched for age and sex was created. The target teeth were the maxillary central incisors (11,21), maxillary lateral incisors (12,22), maxillary canines (13,23), and maxillary first molars (16,26). Dental morphometric variables measured on CBCT images included tooth length, crown height, root length, mesio-distal diameter, crown-to-root ratio, vestibular-palatine diameter, mid mesio-distal diameter, mid buccal-palatal diameter, maximum buccal-palatal diameter, and cervical circumference. The FA2 fluctuating asymmetry index (Palmer and Strobeck, 1986) was applied. Some discrepancies in crown-to-root ratios and root length asymmetry were significantly lower in the DS individuals than in controls. Combining the crown-to-root ratio of tooth 11 versus 21, tooth 12 versus 22, and tooth 13 versus 23, we developed a predictive model with a discriminatory power between DS and controls of 0.983. Some dental morphometric variables may actually be more stable in DS individuals than in the general population. This offers a new perspective on the relationship between canalization, fluctuating asymmetry, and aneuploidy. The work was supported by the Medical-Surgical Dentistry Research Group (OMEQUI), University of Santiago de Compostela (USC), Spain SI

The assessment of vascular biomarkers and their correlation with visual acuity is one of the most important issues in the diagnosis and follow-up of retinal vein occlusions (RVOs). The high workloads of clinical practice make it necessary to have a fast, objective, and automatic method to analyze image features and correlate them with visual function. The aim of this study is to propose a fully automatic system which is capable of estimating visual acuity (VA) in RVO eyes, based only on information obtained from macular optical coherence tomography angiography (OCTA) images. We also propose an automatic methodology to rapidly measure the foveal avascular zone (FAZ) area and the vascular density (VD) in the superficial and deep capillary plexuses in swept-source OCTA images centered on the fovea. The proposed methodology is validated using a representative sample of 133 visits of 50 RVO patients. Our methodology estimates VA with very high precision and is even more accurate when we integrate depth information, providing a high correlation index of 0.869 with the real VA, which outperforms the correlation index of 0.855 obtained when estimating VA from the data obtained by the semiautomatic existing method. In conclusion, the proposed method is the first computational system able to estimate VA in RVO, with the additional benefits of being automatic, less time-consuming, objective and more accurate. Furthermore, the proposed method is able to integrate depth information, a feature which is lacking in the existing method This work has received partial financial support from the Mutua Madrileña project, Ref. 2017/365 SI

Background: To assess the prevalence and severity of caries in 12- and 15-year-old schoolchildren, and to analyse the related risk factors. Methods: We conducted a cross-sectional study on a random sample of 1843 schoolchildren aged 12 and 15 from Galicia (northwest of Spain). Self-administered questionnaire and dental clinical examination were performed to obtain information about oral health habits, dental caries and oral hygiene. A logistic regression model including dental-caries-related variables was generated for each age group. Results: The respective findings for 12- and 15-years-old were as follows: decayed, missing, filled teeth index both for permanent and temporary dentition (DMFT/dmft) of 0.89 (95% CI, 0.87–0.91) and 1.38 (95% CI, 1.33–1.43), respectively; caries prevalence 39.6% (95% CI, 36.3–42.9) and 51.7% (95% CI, 48.0–55.4), respectively. In the 12-yearold group, individuals who occasionally, never or hardly ever brushed their teeth had higher values of caries (OR = 1.83, 95% CI 1.07–3.15, and OR = 9.14, 95% CI1.63–51.17, respectively). Also, the presence of plaque on more than 1/3 gingival was statistically associated with an increase of caries (OR = 2.03; 95% CI, 1.11–3.70), and living in a rural environment was a risk factor (OR = 1.3; 95% CI,1.02–1.80). In the 15-year-old group, higher caries risk was found when brushing was performed once a day (OR = 1.61; 95% CI,1.03–2.50), and among individuals who visited private clinics (OR = 1.77; 95% CI, 1.17–2.66), while electric toothbrush was associated with a lower caries risk (OR = 0.50; 95% CI, 0.29–0.86). Conclusions: This study revealed that risk factors of dental caries showed differences in schoolchildren of 12- and 15-year-old. Strongest evidence related to caries in 12-year-old group were found in frequency of toothbrushing and dental plaque. In 15-year old group, electric toothbrush, time since the last visit to the dentist and type of dental care (public/private) had a stronger association with dental caries. Caries prevalence and mean DMFT/dmft increased from 12- to 15-year-old, in spite of improvement in oral hygiene at the age of 15 This study was authorised and funded by the Galician Regional Health Administration (Consellería de Sanidade, Dirección Xeral de Innovación e Xestión da Saúde Publica, Xunta de Galicia), as coordinated by the Preventive Medicine and Public Health Department of Santiago de Compostela University SI

Background Surgical treatments such as guided tissue regeneration (GTR) and access flap surgery are widely employed for the treatment of intrabony defects. However, little is known regarding the postoperative expression of gingival crevicular fluid (GCF) markers. Objective The aim of this systematic review was to compare the expression of GCF markers following treatment of periodontal intrabony defects with guided tissue regeneration or access surgery. The association of the markers’ expression with the clinical outcome was also assessed. Methods An electronic literature search was conducted in MEDLINE, EMBASE, OpenGrey, LILACS and Cochrane Library up to December 2018 complemented by a manual search. Human, prospective clinical studies were identified. The changes from baseline up to 30 days (early healing) and 3 months (late healing) were assessed. Results A total of 164 publications were identified and reviewed for eligibility. Of these, 10 publications fulfilled the inclusion criteria. The included studies evaluated 15 different GCF markers with a follow-up time between 21 and 360 days postoperatively. PDGF, VEGF and TIMP-1 changes were often investigated in the included studies; however, contrasting results were reported. Two studies agreed that both GTR and OFD lead to similar OPG level changes. TGF-β1 is increased early postoperatively, irrespective of the surgical technique employed. Conclusion There is limited evidence available on the expression of GCF markers after surgical interventions of intrabony periodontal defects. However, OPG and TGF-β1 tend to increase early post-operatively, irrespective of the surgical technique employed, irrespective of the surgical technique employed. Clinical relevance More well-designed, powered studies with sampling periods reflecting the regenerative process are needed, and future research should focus on employing standardised protocols for collecting, storing and analysing GCF markers.

In the last three decades, Clostridium difficile infection (CDI) has increased in incidence and severity in many countries worldwide. The increase in CDI incidence has been particularly apparent among surgical patients. Therefore, prevention of CDI and optimization of management in the surgical patient are paramount. An international multidisciplinary panel of experts from the World Society of Emergency Surgery (WSES) updated its guidelines for management of CDI in surgical patients according to the most recent available literature. The update includes recent changes introduced in the management of this infection SI

Background Accidental falls, especially for the elderly, are a major health issue. Balance disorders are one of their main causes. Vestibular rehabilitation (VR) has proven to be useful in improving balance of elderly patients with instability. Its major handicap is probably its cost, which has prevented its generalisation. So, we have designed a clinical trial with posturographic VR, to assess the optimum number of sessions necessary for a substantial improvement and to compare computerised dynamic posturography (CDP) (visual feedback) and mobile posturography (vibrotactile feedback). Methods Design: randomized controlled trial. It is an experimental study, single-center, open, randomized (balanced blocks of patients) in four branches in parallel, in 220 elderly patients with high risk of falls; follow-up period: twelve months. Setting: Department of Otorhinolaryngology of a tertiary referral hospital. Participants: people over 65 years, fulfilling two or more of the following requirements: a) at least one fall in the last twelve months. b) take at least 16 s or require some support in perform the “timed up and go” test. c) a percentage of average balance in the sensory organization test (SOT) of the CDP 60%. Intervention: Four differents protocols of vestibular rehabilitation (randomization of the patients). Main outcome measure: The percentage of average balance in the SOT-CDP. Secondary measures: time and supports in the “timed up and go” test, scores of the CDP and Vertiguard, and rate of falls. Discussion Posturographic VR has been proven to be useful for improving balance and reducing the number of falls among the aged. However, its elevated cost has limited its use. It is possible to implement two strategies that improve the cost-benefit of posturography. The first involves optimising the number of rehabilitation sessions; the second is based on the use of cheaper posturography systems. Trial registration ClinicalTrials.gov identifier: NCT03034655. Registered on 25 January 2017.

Cancer cells overexpress proton exchangers at the plasma membrane in order acidify the extracellular matrix and maintain the optimal pH for sustaining cancer growth. Among the families of proton exchangers implicated in carcinogenesis, carbonic anhydrases (CAs), monocarboxylate transporters (MCTs), Na+/H+ exchangers (NHEs), sodium bicarbonate cotransporters (NBCs), and vacuolar ATPases (V-ATPases) are highlighted. Considerable research has been carried out into the utility of the understanding of these machineries in the diagnosis and prognosis of several solid tumors. In addition, as therapeutic targets, the interference of their functions has contributed to the discovery or optimization of cancer therapies. According to recent reports, the study of these mechanisms seems promising in the particular case of oral squamous cell carcinoma (OSCC). In the present review, the latest advances in these fields are summarized, in particular, the usefulness of proton exchangers as potential prognostic biomarkers and therapeutic targets in OSCC.

The aim of this study was to quantify the available maxillary alveolar bone in a group of individuals with Down syndrome (DS) to determine the best areas for orthodontic miniscrew placement. The study group consisted of 40 patients with DS aged 12–30 years. We also selected an age and sex-matched control group. All measurements were performed on cross-sectional images obtained with cone-beam computed tomography. The selected areas of interest were the 4 interradicular spaces between the distal wall of the canine and the mesial wall of the second molar, in both maxillary quadrants. We measured the vestibular-palatine (VP) and mesiodistal (MD) dimensions to depths of 3, 6 and 9 mm from the alveolar ridge. We also measured the bone density in the same interradicular spaces of interest to 6 mm of depth from the alveolar crest. VP measurements were longer in the more posterior sectors and as the distance from the alveolar ridge increased. MD measurements also increased progressively as the distance from the alveolar ridge increased. In general, both the VP and MD measurements in the DS group were similar among the male and female participants. As age increased, the MD distance increased, while the VP distance decreased. The VP distance was ≥6 mm in at least 75% of the DS group in practically all assessed interdental spaces. The MD distance was ≥2 mm in at least 75% of the DS group only between the first and second molar, to 9 mm of depth from the alveolar ridge. The safe area for inserting orthodontic miniscrews in DS patients is restricted to the most posterior and deepest area of the maxillary alveolar bone SI

© 2019 by the authors., Tumor-derived extracellular vesicles (EVs) are secreted in large amounts into biological fluids of cancer patients. The analysis of EVs cargoes has been associated with patient´s outcome and response to therapy. However, current technologies for EVs isolation are tedious and low cost-efficient for routine clinical implementation. To explore the clinical value of circulating EVs analysis we attempted a proof-of-concept in endometrial cancer (EC) with ExoGAG, an easy to use and highly efficient new technology to enrich EVs. Technical performance was first evaluated using EVs secreted by Hec1A cells. Then, the clinical value of this strategy was questioned by analyzing the levels of two well-known tissue biomarkers in EC, L1 cell adhesion molecule (L1CAM) and Annexin A2 (ANXA2), in EVs purified from plasma in a cohort of 41 EC patients and 20 healthy controls. The results demonstrated the specific content of ANXA2 in the purified EVs fraction, with an accurate sensitivity and specificity for EC diagnosis. Importantly, high ANXA2 levels in circulating EVs were associated with high risk of recurrence and non-endometrioid histology suggesting a potential value as a prognostic biomarker in EC. These results also confirmed ExoGAG technology as a robust technique for the clinical implementation of circulating EVs analyses., This research was funded by Instituto de Salud Carlos III, grant PI17/01919, co-financed by the European Regional Development Fund (FEDER), and by Fundación Científica de la Asociación Española Contra el Cáncer (AECC), Grupos Clínicos Coordinados 2018. Carolina Herrero is supported by a predoctoral i-PFIS fellowship from Instituto de Salud Carlos III (IFI17/00047); Laura Muinelo is supported by Asociación Española Contra el Cáncer (AECC).

Oral cavity cancer is the most frequent malignancy of the head and neck. Unfortunately, despite educational interventions for prevention and early diagnosis, oral cancer patients are often diagnosed in advanced stages associated with poor prognosis and life expectancy. Therefore, there is an urgent need to find noninvasive biomarkers to improve early detection of this tumor. Liquid biopsy has emerged as a valuable tool in medical oncology which provides new horizons for improving clinical decision making. Notably, cell-free microRNAs (miRNAs), a class of short non-coding RNAs, are emerging as novel noninvasive cancer biomarkers. Here, we provide an overview of the potential clinical application of cell-free miRNAs as diagnostic, prognostic, and therapeutic biomarkers in oral cancer SI

Background: To evaluate the relationship between the plasmatic CD63 and CAV1 positive exosome levels, in patients with OSCC before and after surgical treatment and to correlate it with their overall survival. Methods: A double-blind pilot study over 10 patients OSCC and T4 stage without distant metastases or local bone invasion has been performed. The average follow-up period was 37.64 months (34.3&ndash, 40.84). We obtained 2 plasma tubes of 1 mL each before surgery and 7 days after surgery. Before performing the immunocapture-based analysis, EVs (Extracellular Vesicles) were isolated from the plasma and characterized with western blot analysis. Results: Mean values of CD63 positive plasmatic exosomes (EXO-CD63) after surgery decreased from 750.88 &plusmn, 286.67 to 541.71 &plusmn, 244.93 (p = 0.091). On the other hand, CAV-1 positive plasmatic exosomes (EXO-CAV-1) increased after surgery from 507 &plusmn, 483.39 to 1120.25 &plusmn, 1151.17 (p = 0.237). Patients with EXO-CD63 levels lower than the mean global value before the surgery had a survival of 36.04 months compared with the group with EXO-CD63 higher than the average who only survived 12.49 &plusmn, 1.67 months from the diagnosis, p = 0.225. When EXO-CAV-1 levels before surgery was lower than the average (813.94 &plusmn, 801.21) overall survival was 24.69 &plusmn, 22.23 months in contrast when it was higher that was only 11.64 months, p = 0.157. Patients with lower EXO-CD63 levels after surgery lived an average of 23.84 &plusmn, 23.9 months, while those with higher plasmatic levels of EXO-CD63 live 13.35 months, p = 0.808. When EXO-CAV-1 levels after surgery were lower, the average overall survival was 20.344 &plusmn, 15.40 months, in contrast when the EXO-CAV-1 levels were higher showing rather an estimate survival expectation of 1.64 months. Conclusions: Surgical treatment induced a dramatic reduction of the plasmatic levels of exosomes expressing CD63 as early as 1 week after resection. This first result suggests that the tumour mass is responsible of the high levels of circulating exosomes detected in cancer patients. At the same time point exosome expressing CAV-1 increased, possibly due to the inflammatory reaction immediately after surgery. Lastly, statistical analysis showed that lower levels of plasmatic exosomes both before and after surgery correlated with a better life expectancy of OSCC patients. Hopefully, this approach will prove useful in the clinical follow-up of cancer patients.

Combined central retinal artery and vein occlusion is an uncommon vascular pathology that can cause severe and permanent visual impairment. Optical coherence tomography angiography (OCTA) is a newly available, noninvasive imaging technique that can potentially improve understanding of the structural and vascular implications and prognosis of this infrequent pathology. The present report describes the principal clinical findings in a case of combined central retinal artery and vein occlusion, as detected by the different imaging modalities available in a tertiary referral hospital. OCTA wide-field montage images identified an extensive area of nonperfusion on the macula with involvement of the entire retina at nearly 360°. We observed the most severe nonperfusion in the deep capillary plexus, while perfusion of the choriocapillaris was unaffected. Meanwhile, fluorescein angiography (FA) findings revealed a delay in perfusion rate with marked nonperfusion areas in the peripheral retina at 360°. We identified that the wide-field OCTA montage permitted visualization of a similar or wider peripheral retinal area compared with FA. Therefore, OCTA is potentially useful for assessment of the global retinal nonperfusion status at baseline and during follow-up, with the added advantage of being a noninvasive technique This work was supported in part by the Mutua Madrileña project (Ref. 2017/365) SI

The aim of this present study was to comprehensively describe somatic DNA alterations and transcriptional alterations in the last extension of the HNSCC subsets in TCGA, encompassing a total of 528 tumours. In order to achieve this goal, transcriptional analysis, functional enrichment assays, survival analysis, somatic copy number alteration analysis and somatic alteration analysis were carried out. A total of 3491 deregulated genes were found in HNSCC patients, and the functional analysis carried out determined that tissue development and cell differentiation were the most relevant signalling pathways in upregulated and downregulated genes, respectively. Somatic copy number alteration analysis showed a &ldquo, top five&rdquo, altered HNSCC genes: CDKN2A (deleted in 32.03% of patients), CDKN2B (deleted in 28.34% of patients), PPFIA1 (amplified in 26.02% of patients), FADD (amplified in 25.63% of patients) and ANO1 (amplified in 25.44% of patients). Somatic mutations analysis revealed TP53 mutation in 72% of the tumour samples followed by TTN (39%), FAT1 (23%) and MUC16 (19%). Another interesting result is the mutual exclusivity pattern that was discovered between the TP53 and PIK3CA mutations, and the co-occurrence of CDKN2A with the TP53 and FAT1 alterations. On analysis to relate differential expression genes and somatic copy number alterations, some genes were overexpressed and amplified, for example, FOXL2, but other deleted genes also showed overexpression, such as CDKN2A. Survival analysis revealed that overexpression of some oncogenes, such as EGFR, CDK6 or CDK4 were associated with poorer prognosis tumours. These new findings help us to develop new therapies and programs for the prevention of HNSCC.

The aim of this study was to evaluate the potential anti-biofilm and antibacterial activities of Streptococcus downii sp. nov. To test anti-biofilm properties, Streptococcus mutans, Actinomyces naeslundii, Veillonella parvula, Fusobacterium nucleatum, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans were grown in a biofilm model in the presence or not of S. downii sp. nov. for up to 120 h. For the potential antibacterial activity, 24 h-biofilms were exposed to S. downii sp. nov for 24 and 48 h. Biofilms structures and bacterial viability were studied by microscopy, and the effect in bacterial load by quantitative polymerase chain reaction. A generalized linear model was constructed, and results were considered as statistically significant at p &lt, 0.05. The presence of S. downii sp. nov. during biofilm development did not affect the structure of the community, but an anti-biofilm effect against S. mutans was observed (p &lt, 0.001, after 96 and 120 h). For antibacterial activity, after 24 h of exposure to S. downii sp. nov., counts of S. mutans (p = 0.019) and A. actinomycetemcomitans (p = 0.020) were significantly reduced in well-structured biofilms. Although moderate, anti-biofilm and antibacterial activities of S. downii sp. nov. against oral bacteria, including some periodontal pathogens, were demonstrated in an in vitro biofilm model.

Objective: To describe the subgingival microbiome of individuals with Down syndrome (DS). Methods: We conducted a cross-sectional observational study that obtained bacterial DNA samples from 50 patients with DS, 25 with periodontitis (PDS) and 25 with a healthy periodontal condition (HDS). The samples were analyzed by sequencing the 16S rRNA gene V3&ndash, V4 hypervariable region using the MiSeq System. Taxonomic affiliations were assigned using the na&iuml, ve Bayesian classifier integrated in QIIME2 plugins. We evaluated the difference in bacteria abundance between the sample groups using Wilcoxon and Kruskal&ndash, Wallis tests. We evaluated the alpha diversity of the identified species using the Observed, Chao1metric, ACE and Shannon indices and evaluated beta diversity with principal coordinate analysis (registration code: 2018/510). Results: Twenty-one genera and 39 bacterial species showed a significantly different abundance between the study groups. Among the genera, Porphyromonas, Treponema, Tannerella and Aggregatibacter were more abundant in the PDS group than in the HDS group, as were the less commonly studied Filifactor, Fretibacterium and Desulfobulbus genera. Among the species, Porphyromonas spp. and Tannerella spp. were the most abundant in the PDS group, the most abundant species in the HDS group were Pseudomonas spp., Granulicatella spp. and Gemella spp. Conclusion: Well-recognized periodontal pathogens and newly proposed pathogenic taxa were associated with periodontitis in patients with DS.

Càncer de pròstata resistent a la castració (CRPC); Cèl·lules tumorals circulants (CTC); Marcadors tumorals Cáncer de próstata resistente a la castración (CRPC); Células tumorales circulantes (CTC); Marcadores tumorales Castration-resistant prostate cancer (CRPC); Circulating tumor cells (CTCs); Tumor markers Background: Current therapeutic options in the course of metastatic castration-resistant prostate cancers (mCRPC) reinforce the need for reliable tools to characterize the tumor in a dynamic way. Circulating tumor cells (CTCs) have emerged as a viable solution to the problem, whereby patients with a variety of solid tumors, including PC, often do not have recent tumor tissue available for analysis. The biomarker characterization in CTCs could provide insights into the current state of the disease and an overall picture of the intra-tumor heterogeneity. Methods: in the present study, we applied a global gene expression characterization of the CTC population from mCRPC (n = 9), with the goal to better understand the biology of these cells and identify the relevant molecules favoring this tumor progression. Results: This analysis allowed the identification of 50 genes specifically expressed in CTCs from patients. Six of these markers (HOXB13, QKI, MAOA, MOSPD1, SDK1, and FGD4), were validated in a cohort of 28 mCRPC, showing clinical interest for the management of these patients. Of note, the activity of this CTC signature was related to the regulation of MYC, a gene strongly implicated in the biology of mCRPC. Conclusions: Overall, our results represent new evidence on the great value of CTCs as a non-invasive biopsy to characterize PC. This work was partially financed with the “liquid Biopsy crowdfunding, 2017”. L.M-L. is supported by AECC.

The objective of the present study was to determine cytokine thresholds derived from predictive models for the diagnosis of chronic periodontitis, differentiating by smoking status. Seventy-five periodontally healthy controls and 75 subjects affected by chronic periodontitis were recruited. Sixteen mediators were measured in gingival crevicular fluid (GCF) using multiplexed bead immunoassays. The models were obtained using binary logistic regression, distinguishing between non-smokers and smokers. The area under the curve (AUC) and numerous classification measures were obtained. Model curves were constructed graphically and the cytokine thresholds calculated for the values of maximum accuracy (ACC). There were three cytokine-based models and three cytokine ratio-based models, which presented with a bias-corrected AUC > 0.91 and > 0.83, respectively. These models were (cytokine thresholds in pg/ml for the median ACC using bootstrapping for smokers and non-smokers): IL1alpha (46099 and 65644); IL1beta (4732 and 5827); IL17A (11.03 and 17.13); IL1alpha/IL2 (4210 and 7118); IL1beta/IL2 (260 and 628); and IL17A/IL2 (0.810 and 1.919). IL1alpha, IL1beta and IL17A, and their ratios with IL2, are excellent diagnostic biomarkers in GCF for distinguishing periodontitis patients from periodontally healthy individuals. Cytokine thresholds in GCF with diagnostic potential are defined, showing that smokers have lower threshold values than non-smokers. This work was supported by the Instituto de Salud Carlos III (General Division of Evaluation and Research Promotion, Madrid, Spain) and co-financed by FEDER (“A way of making Europe”) under Grant ISCIII/PI17/01722, and the Consellería de Cultura, Educación e Ordenación Universitaria da Xunta de Galicia (Spain) under Grant ED431B 2017/029 and A. Regueira-Iglesias support ED481A-2017 SI

This study aimed to compare the bacterial viability and diversity of a substrate-formed biofilm (SF-biofilm) in situ to a supragingival tooth-formed biofilm (TF-biofilm) in the same group of individuals. The impact of the device/disc position and toothbrushing during the formation of SF-biofilm was also assessed. Two tests were run. In test 1, 15 volunteers wore two hemi-splints carrying six discs of human enamel, glass, and hydroxyapatite for 2 days, and were instructed to not perform any oral hygiene measure. Biofilm samples were collected from the substrates and the contralateral tooth and were analysed using CLSM. In five volunteers, half of the biofilm present on the discs and their contralateral teeth were scraped and analysed using 16S pyrosequencing. In test 2, the microscopic analysis was repeated only on the SF-biofilm samples, and the volunteers were allowed to brush their teeth. Multivariate analyses revealed that the donors had a significant effect on the composition of the biofilm, confirming its subject-dependent character. The bacterial composition of the SF-biofilm was similar to the TF-biofilm, with significant differential abundance detected in very few taxa of low abundance. The toothbrushing during the formation of SF-biofilm was the only factor that conditioned the thickness or bacterial viability This work was supported by the Instituto de Salud Carlos III (General Division of Evaluation and Research Promotion, Madrid, Spain) and co-financed by FEDER (‘A way of making Europe’) under Grant ISCIII/PI17/ 01722 and the Consellería de Cultura, Educación e Ordenación Universitaria da Xunta de Galicia (Spain) under Grant ED431B 2017/029. Specifically, the development of the Matlab toolbox called the Dentius Biofilm received financial support from the Consellería de Cultura, Educación e Ordenación Universitaria (accreditation 2016-2019) under Grant ED431G/08 SI

Objectives Studies on dental plaque removal by chewing an apple are scarce and dated, with conflicting findings. This study aimed to determine whether chewing an apple produced mechanical removal of dental plaque or had any effect on salivary bacterial viability. Methods The study group consisted of 20 healthy adults with good oral health status who were randomly assigned to brush their teeth or eat an apple. After 2 weeks, the experiment was repeated with the order reversed. Plaque index (PI) and the bacterial viability (BV) in a sample of whole saliva (spit) were determined before brushing or apple eating (baseline, B), immediately afterward (A) and 24 hours afterward (24). Results After chewing an apple, PI-A was significantly higher than both PI-B (P < .001) and PI-24 (P < .001). BV-A was significantly lower than BV-B (P < .001), with a return to baseline values at the BV-24 measurement. Conclusions Chewing an apple does not remove dental plaque, and may favor plaque regrowth during the first 24 hours, but it does produce an immediate reduction in salivary bacterial viability similar to that after tooth brushing SI

La queilitis actinica (QA) es una patologia que afecta principalmente al bermellon del labio inferior dando lugar a alteraciones clinicas y patologicas. La QA se considera mayoritariamente una forma superficial de carcinoma espinocelular y el principal factor de riesgo en su desarrollo es la exposicion solar cronica. La epidemiologia de la QA ha sido escasamente analizada en el pasado. Por este motivo, se diseno un estudio multicentrico transversal en la poblacion de la comunidad autonoma de Galicia con edad igual o superior a 45 anos para evaluar la prevalencia de QA, analizar sus factores de riesgo y manifestaciones clinicas e investigar el uso de fotoproteccion especifica en los labios. Se incluyeron 1250 pacientes en el estudio y de ellos, 410 presentaron QA. La prevalencia de QA en nuestro estudio fue 31,3%. Los sujetos con edad igual o superior a 60 anos, con fototipo II de Fitzpatrick, que habian trabajado en exteriores durante mas de 25 anos y con historia previa de cancer cutaneo no melanoma tuvieron un riesgo superior de desarrollar QA. La mayoria de pacientes con QA no realizaban fotoproteccion labial y, cuando lo hicieron, esta fue insuficiente; solo los pacientes con QA y fototipo I y II de Fitzpatrick mostraron tendencia al uso de fotoproteccion labial. La presencia de otros signos de dano actinico cronico, como queratosis actinicas y/o lentigos solares, fueron prevalentes en los pacientes con QA. En vista de los resultados, es necesario que se promuevan medidas educativas que insistan en la importancia de la fotoproteccion labial, especialmente en poblaciones de alto riesgo; ademas, la exploracion labial minuciosa deberia ser obligatoria en todos los pacientes con dano actinico cronico.

Class III malocclusions are considered to be one of the most difficult problems to treat. Due to the significant number of patients with Class III malocclusion who cannot undergo orthognathic surgery for different reasons, we have proposed an alternative treatment that we have called surgically assisted rapid palatal expansion (SARPE) + temporary anchorage devices (TADs) which allows solving mild and moderate Class III malocclusion combined with maxillary compression, obtaining acceptable esthetic and functional results. We present a case report of an adult female with skeletal Class III malocclusion with compression in the maxillary and mandibular asymmetry, who was treated with SARPE + TADs. The result is acceptable in terms of occlusion function, esthetic of the smile, and facial esthetics SI

Osteoarthritis (OA), a chronic disease characterized by articular cartilage degeneration, is a leading cause of disability and pain worldwide. In OA, chondrocytes in cartilage undergo phenotypic changes and senescence, restricting cartilage regeneration and favouring disease progression. Similar to other wound-healing disorders, chondrocytes from OA patients show a chronic increase in the gap junction channel protein connexin43 (Cx43), which regulates signal transduction through the exchange of elements or recruitment/release of signalling factors. Although immature or stem-like cells are present in cartilage from OA patients, their origin and role in disease progression are unknown. In this study, we found that Cx43 acts as a positive regulator of chondrocyte-mesenchymal transition. Overactive Cx43 largely maintains the immature phenotype by increasing nuclear translocation of Twist-1 and tissue remodelling and proinflammatory agents, such as MMPs and IL-1β, which in turn cause cellular senescence through upregulation of p53, p16INK4a and NF-κB, contributing to the senescence-associated secretory phenotype (SASP). Downregulation of either Cx43 by CRISPR/Cas9 or Cx43-mediated gap junctional intercellular communication (GJIC) by carbenoxolone treatment triggered rediferentiation of osteoarthritic chondrocytes into a more differentiated state, associated with decreased synthesis of MMPs and proinflammatory factors, and reduced senescence. We have identified causal Cx43- sensitive circuit in chondrocytes that regulates dedifferentiation, redifferentiation and senescence. We propose that chondrocytes undergo chondrocyte-mesenchymal transition where increased Cx43-mediated GJIC during OA facilitates Twist-1 nuclear translocation as a novel mechanism involved in OA progression. These findings support the use of Cx43 as an appropriate therapeutic target to halt OA progression and to promote cartilage regeneration. This work was supported in part through funding from the Spanish Society for Rheumatology (SER; FER 2013) and Spanish Foundation for Research on Bone and Mineral Metabolism (FEIOMM), grant PRECIPITA-2015-000139 from the FECYT-Ministry of Economy and Competitiveness (to M.D.M.), grant PI16/00035 from the Health Institute ‘Carlos III’ (ISCIII, Spain), the European Regional Development Fund, ‘A way of making Europe’ from the European Union (to M.D.M.) and a grant from Xunta de Galicia IN607B 2017/21 (to M.D.M.) and pre-doctoral fellowship to M.V.-E. T.A. acknowledges support from Instituto de Salud Carlos III grants PI16/00772 and CPII16/00042, co-financed by the European Regional Development Fund (ERDF) SI

Actinic cheilitis (AC) is a common condition that mainly involves the lower lip, which is associated with chronic exposure to ultraviolet (UV) radiation. AC is considered a precursor of malignancy (1), but the rate of progression from AC to invasive squamous cell carcinoma (SCC) has not yet been established. An epidemiological study previously described the prevalence of AC and its associated variables in the Galicia region (north-western Spain); the prevalence of AC in a population aged 45 years and over was 31.3%, and multivariate analysis showed that significant and independent risk factors for AC were age ≥60 years, Fitzpatrick skin phototypes I and II, working outdoors for more than 25 years, and a history of non-melanoma skin cancer (NMSC) (2). We report here a subanalysis of the clinical manifestations of AC and the associations of AC with other markers of actinic damage SI

Currently, there is little evidence on the in situ antibacterial activity of essential oils (EO) without alcohol. This study aimed to evaluate in situ the substantivity and antiplaque effect on the plaque-like biofilm (PL-biofilm) of two solutions, a traditional formulation that contains EO with alcohol (T-EO) and an alcohol-free formulation of EO (Af-EO). Eighteen healthy adults performed a single mouthwash of: T-EO, Af-EO, and sterile water (WATER) after wearing an individualized disk-holding splint for 2 days. The bacterial viability (BV) and thickness of the PL-biofilm were quantified at baseline, 30 s, and 1, 3, 5, and 7 h post-rinsing (Test 1). Subsequently, each volunteer wore the splint for 4 days, applying two daily mouthwashes of: T-EO, Af-EO, and WATER. The BV, thickness, and covering grade (CG) of the PL-biofilm were quantified (Test 2). Samples were analyzed by confocal laser scanning microscopy after staining with the LIVE/DEAD® BacLight™ solution. To conduct the computations of the BV automatically, a Matlab toolbox called Dentius Biofilm was developed. In test 1, both EO antiseptics had a similar antibacterial effect, reducing BV after a single rinse compared to the WATER, and keeping it below baseline levels up to 7 h post-rinse (P < 0.001). The mean thickness of the PL-biofilm after rinsing was not affected by any of the EO formulations and ranged from 18.58 to 20.19 μm. After 4 days, the T-EO and Af-EO solutions were significantly more effective than the WATER, reducing the BV, thickness, and CG of the PL-biofilm (P < 0.001). Although, both EO antiseptics presented a similar bactericidal activity, the Af-EO rinses led to more significant reductions in the thickness and CG of the PL-biofilm than the T-EO rinses (thickness = 7.90 vs. 9.92 μm, P = 0.012; CG = 33.36 vs. 46.61%, P = 0.001). In conclusion, both essential oils antiseptics had very high immediate antibacterial activity and substantivity in situ on the 2-day PL-biofilm after a single mouthwash. In the 4-day PL-biofilm, both essential oils formulations demonstrated a very good antiplaque effect in situ, although the alcohol-free formula performed better at reducing the biofilm thickness and covering grade This investigation was supported by the 2014-CE2010 project financed by Johnson & Johnson Company, Madrid, Spain. Specifically, the development of the Matlab toolbox called Dentius Biofilm received financial support from the Consellería de Cultura, Educación e Ordenación Universitaria (accreditation 2016-2019, ED431G/08) and the European Regional Development Fund (ERDF) SI

Although a distinct cytokine profile has been described in the gingival crevicular fluid (GCF) of patients with chronic periodontitis, there is no evidence of GCF cytokine-based predictive models being used to diagnose the disease. Our objectives were: to obtain GCF cytokine-based predictive models; and develop nomograms derived from them. A sample of 150 participants was recruited: 75 periodontally healthy controls and 75 subjects affected by chronic periodontitis. Sixteen mediators were measured in GCF using the Luminex 100™ instrument: GMCSF, IFNgamma, IL1alpha, IL1beta, IL2, IL3, IL4, IL5, IL6, IL10, IL12p40, IL12p70, IL13, IL17A, IL17F and TNFalpha. Cytokine-based models were obtained using multivariate binary logistic regression. Models were selected for their ability to predict chronic periodontitis, considering the different role of the cytokines involved in the inflammatory process. The outstanding predictive accuracy of the resulting smoking-adjusted models showed that IL1alpha, IL1beta and IL17A in GCF are very good biomarkers for distinguishing patients with chronic periodontitis from periodontally healthy individuals. The predictive ability of these pro-inflammatory cytokines was increased by incorporating IFN gamma and IL10. The nomograms revealed the amount of periodontitis-associated imbalances between these cytokines with pro-inflammatory and anti-inflammatory effects in terms of a particular probability of having chronic periodontitis This work was supported by project EM2014/025 from Regional Ministry of Culture, Education and University (regional government of Galicia, Spain), which is integrated in Regional Plan of Research, Innovation and Development 2011–2015 SI

Currently, there is little evidence available on the development of predictive models for the diagnosis or prognosis of chronic periodontitis based on the qPCR quantification of subgingival pathobionts. Our objectives were to: (1) analyze and internally validate pathobiont-based models that could be used to distinguish different periodontal conditions at site-specific level within the same patient with chronic periodontitis; (2) develop nomograms derived from predictive models. Subgingival plaque samples were obtained from control and periodontal sites (probing pocket depth and clinical attachment loss 4 mm, respectively) from 40 patients with moderate-severe generalized chronic periodontitis. The samples were analyzed by qPCR using TaqMan probes and specific primers to determine the concentrations of Actinobacillus actinomycetemcomitans (Aa), Fusobacterium nucleatum (Fn), Parvimonas micra (Pm), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Tannerella forsythia (Tf), and Treponema denticola (Td). The pathobiont-based models were obtained using multivariate binary logistic regression. The best models were selected according to specified criteria. The discrimination was assessed using receiver operating characteristic curves and numerous classification measures were thus obtained. The nomograms were built based on the best predictive models. Eight bacterial cluster-based models showed an area under the curve (AUC) ≥0.760 and a sensitivity and specificity ≥75.0%. The PiTfFn cluster showed an AUC of 0.773 (sensitivity and specificity = 75.0%). When Pm and AaPm were incorporated in the TdPiTfFn cluster, we detected the two best predictive models with an AUC of 0.788 and 0.789, respectively (sensitivity and specificity = 77.5%). The TdPiTfAa cluster had an AUC of 0.785 (sensitivity and specificity = 75.0%). When Pm was incorporated in this cluster, a new predictive model appeared with better AUC and specificity values (0.787 and 80.0%, respectively). Distinct clusters formed by species with different etiopathogenic role (belonging to different Socransky’s complexes) had a good predictive accuracy for distinguishing a site with periodontal destruction in a periodontal patient. The predictive clusters with the lowest number of bacteria were PiTfFn and TdPiTfAa, while TdPiTfAaFnPm had the highest number. In all the developed nomograms, high concentrations of these clusters were associated with an increased probability of having a periodontal site in a patient with chronic periodontitis This work was supported by the EM2014/025 project from the Regional Ministry of Culture, Education and University (regional government of Galicia, Spain), which is integrated in the Regional Plan of Research, Innovation and Development 2011–2015; and grants PI13/02390-PI16/01163 awarded to MT within the State Plan for R+D+I 2013–2016 (National Plan for Scientific Research, Technological Development and Innovation 2008–2011) and co-financed by the ISCIII-Deputy General Directorate of evaluation and Promotion of Research-European Regional Development Fund “A way of Making Europe” and Instituto de Salud Carlos III FEDER SI