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Detection of COVID-19 manifestations in lung ultrasound (US) images has gained attention in recent times. The current state-of-the-art technique for distinguishing a healthy lung from COVID-19 infected or bacterial pneumonia infected lung uses non-adjacent frames or equally spaced frames from the video. However, the frame content or correlation between the selected frames has not been taken into consideration for frame selection. In this paper, a metric-based frame selection approach is proposed for three-way classification of lung US videos, and the influence of the frame selection method on image classification accuracy is studied. A deep learning model comprising of a pre-trained model (VGG16) for feature extraction, multi-head attention for feature calibration, global averaging for feature reduction, and a dense layer for classification is proposed. The pre-trained model is re-trained using cross-entropy loss with balanced weights to handle class imbalance. Two types of classification approaches are considered: i) few frames in a video are selected using the proposed metrics; and (ii) all frames in a video are considered. With VGG16 as the pre-trained model, a mean balanced sensitivity of COVID-19, bacterial pneumonia, and healthy classes with 0.82, 0.89, and 0.87, respectively was achieved using 5-fold cross-validation. The results show that even random selection of frames performs better than fixed frame selection and the proposed frame selection method outperforms the state-of-art fixed frame selection irrespective of the type of backbone model used for lung US classification.

Abstract Previously, in small scale demonstrations, researchers have increased photovoltaic efficiency through cooling by enhancing heat transfer from panels to the air through wind speed. Here I show in the real-world operation of a larger scale photovoltaic generator that increases in wind speed can lead to small but notable energy losses, reflected in the mismatch losses directly derived from the operating voltage of each module. Temperature distribution was measured simultaneously with the operating voltages, alongside the local wind speed and direction. Temperature differences arose from the variable heat transfer throughout the panel, depending on the wind incidence. This affected the operating temperature of each module, consequently affecting their operating voltage and the overall mismatch losses with losses increasing by up to 0.28%. My results suggest that wind patterns cannot be neglected, considering long-term energy estimations and the lifespan of a photovoltaic power plant.

ObjectiveAlthough there have been remarkable achievements in the molecular landscape of oral squamous cell carcinoma (OSCC) in recent years, bringing advances in the understanding of its pathogenesis, development and progression, little has been applied in the prognosis and choosing the optimal treatment. In this study, we explored the influence of the stress induced phosphoprotein 1 (STIP1), which is frequently reported to be highly expressed in many cancers, in OSCCs.MethodsSTIP1 expression was assessed in the TCGA database and in two independent cohorts by immunohistochemistry. Knockdown strategy was applied in OSCC cell lines to determine the impact of STIP1 on viability, proliferation, migration and invasion. The zebrafish model was applied for studying tumor formation and metastasis in vivo. The association of STIP1 and miR-218-5p was explored by bioinformatics and mimics transfection.ResultsSTIP1 was highly expressed in OSCCs and significantly associated with shortened survival and higher risk of recurrence. STIP1 down-regulation decreased proliferation, migration and invasion of tumor cells, and reduced the number of metastases in the Zebrafish model. STIP1 and miR-218-5p were inversely expressed, and the transfection of miR-218-5p mimics into OSCC cells decreased STIP1 levels as well as proliferation, migration and invasion.ConclusionOur findings show that STIP1 overexpression, which is inversely associated with miR-218-5p levels, contributes to OSCC aggressiveness by controlling proliferation, migration and invasion and is a determinant of poor prognosis.

Understanding macrophage biology can improve comprehension of diverse biological processes and provide insights into novel therapeutic immunomodulatory strategies. Due to limited yield and technical difficulty in isolating primary macrophages, in vitro studies commonly use monocytes as precursor cells. Monocytic cell lines are a virtually unlimited source of macrophage precursors and two of the most frequently used cell lines are THP-1 and U937. Besides a great variability in macrophage differentiation protocols there is scarce information on possible differences in the biological responses of these cell lines. In this study, we used a standardized differentiation protocol using PMA and compared the response of macrophages derived from THP-1 and U937 cells to M1-and M2-polarizing conditions. THP-1-derived macrophages are more responsive to M1 stimuli and skewed towards M1 phenotype, whereas U937-derived macrophages were more responsive to M2 stimuli and skewed towards M2 phenotype. THP-1-derived macrophages also had greater production of ROS and phagocytic activity. Under M1-polarizing conditions, macrophages derived from both THP-1 and U937 reduced phagocytosis activity and the increased production of ROS. This information should be considered to make an informed choice on the cell line used as in vitro macrophage model, according to the experimental goals and biological context.

Abstract: Modified formulations of calcium silicate repair materials with additives have been developed to enhance handling, consistency, biocompatibility and bioactivity. Considering the relevance of osteoblastic cell response to mineralized tissue repair, human osteoblastic cells (Saos-2 cells overexpressing BMP-2) were exposed to mineral trioxide aggregate (MTA) (with calcium tungstate - CaWO4), MTA HP Repair, Bio-C Repair and Bio-C Pulpo. Cell viability was assessed by 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) and neutral red (NR), and cell death, by flow cytometry. Gene expression of bone morphogenetic protein 2 (BMP-2), runt-related transcription factor 2 (RUNX-2), and alkaline phosphatase (ALP) osteogenic markers were evaluated by real-time polymerase chain reaction (RT-qPCR). ALP activity and alizarin red staining (ARS) were used to detect mineralization nodule deposition. Bioactive cements presented no cytotoxic effect, and did not induce apoptosis at the higher dilution (1:12). MTA, Bio-C Repair and Bio-C Pulpo exhibited higher ALP activity than the control group (P < 0.05) after 7 days. MTA, MTA HP and Bio-C Pulpo affected the formation of mineralized nodules (p < 0.05). Exposure to all cement extracts for 1 day increased BMP-2 gene expression. RUNX-2 mRNA was greater in MTA, MTA HP and Bio-C Repair. MTA, MTA HP and Bio-C Pulpo increased the ALP mRNA expression, compared with BMP-2 unexposed cells (P < 0.05). Calcium silicate cements showed osteogenic potential and biocompatibility in Saos-2 cells transfected BMP-2, and increased the mRNA expression of BMP-2, RUNX-2, and ALP osteogenic markers in the BMP-2 transfected system, thereby promoting a cellular response to undertake the mineralized tissue repair.

High dose rate brachytherapy (HDR) is an internal based radiation treatment for prostate cancer. The treatment can deliver radiation to the site of dominant tumor growth within the prostate. Imaging methods to delineate the dominant tumor are imperative to ensure the maximum success of HDR. This paper investigates the feasibility of using electrical impedance tomography (EIT) as the main imaging modality during robot-aided internal radiation therapy. A procedure utilizing brachytherapy needles in order to perform EIT for the purpose of robot-aided prostate cancer imaging is proposed. It is known that cancerous tissue exhibits different conductivity than healthy tissue. Using this information, it is hypothesized that a conductivity map of the tissue can be used to locate and delineate cancerous nodules via EIT. Multiple experiments were conducted using eight brachytherapy needle electrodes. Observations indicate that the imaging procedure is able to observe differences in tissue conductivity in a setting that approximates transperineal HDR and confirm that brachytherapy needles can be used as electrodes for this purpose. The needles can access the tissue at a specific depth that traditional EIT surface electrodes cannot. The results indicate the feasibility of using brachytherapy needles for EIT for the purpose internal radiation therapy.

The sustained increase in new cases of COVID-19 across the world and potential for subsequent outbreaks call for new tools to assist health professionals with early diagnosis and patient monitoring. Growing evidence around the world is showing that lung ultrasound examination can detect manifestations of COVID-19 infection. Ultrasound imaging has several characteristics that make it ideally suited for routine use: small hand-held systems can be contained inside a protective sheath, making it easier to disinfect than X-ray or computed tomography equipment; lung ultrasound allows triage of patients in long term care homes, tents or other areas outside of the hospital where other imaging modalities are not available; and it can determine lung involvement during the early phases of the disease and monitor affected patients at bedside on a daily basis. However, some challenges still remain with routine use of lung ultrasound. Namely, current examination practices and image interpretation are quite challenging, especially for unspecialized personnel. This paper reviews how lung ultrasound (LUS) imaging can be used for COVID-19 diagnosis and explores different image processing methods that have the potential to detect manifestations of COVID-19 in LUS images. Then, the paper reviews how general lung ultrasound examinations are performed before addressing how COVID-19 manifests itself in the images. This will provide the basis to study contemporary methods for both segmentation and classification of lung ultrasound images. The paper concludes with a discussion regarding practical considerations of lung ultrasound image processing use and draws parallels between different methods to allow researchers to decide which particular method may be best considering their needs. With the deficit of trained sonographers who are working to diagnose the thousands of people afflicted by COVID-19, a partially or totally automated lung ultrasound detection and diagnosis tool would be a major asset to fight the pandemic at the front lines.

Background. Type 2 diabetes mellitus (T2DM) and periodontitis (P) commonly occur as comorbidities, but the commonalities in the genetic makeup of affected individuals is largely unknown. Since dyslipidemia is a frequent condition in these individuals, we investigate the association of genomic variations in genes involved in lipid metabolism with periodontal, glycemic, lipid profiles, and the association with periodontitis and T2DM (as comorbidities). Methods. Based on clinical periodontal examination and biochemical evaluation, 893 subjects were divided into T2DM+P (T2DM subjects also affected by periodontitis, n=205), periodontitis (n=345), and healthy (n=343). Fourteen single-nucleotide polymorphisms (SNPs) were investigated: LDLR gene (rs5925 and rs688), APOB (rs676210, rs1042031, and rs693), ABCC8 (rs6544718 and 6544713), LPL (rs28524, rs3735964, and rs1370225), HNF1A (rs2650000), APOE (rs429358 and rs7412), and HNF4A (rs1800961). Multiple linear and logistic regressions (adjusted for covariates) were made for all populations and stratified by sex and smoking habits. Results. Individuals carrying APOB-rs1042031-CT (mainly women and never smokers) had a lower risk of developing periodontitis and T2DM (T2DM+P); altogether, this genotype was related with healthier glycemic, lipid, and periodontal parameters. Significant disease-phenotype associations with gene-sex interaction were also found for carriers of APOB-rs1676210-AG, HNF4A-rs1800961-CT, ABCC8-rs6544718-CT, LPL-rs13702-CC, and LPL-rs285-CT. Conclusions. Polymorphisms in lipid metabolism genes are associated with susceptibility to T2DM-periodontitis comorbidities, demonstrating gene-sex interaction. The APOB-rs1042031 was the most relevant gene marker related to glucose and lipid metabolism profiles, as well as with obesity and periodontitis.

Percutaneous Nephrolithotomy is the standard surgical procedure used to remove large kidney stones. PCNL procedures have a steep learning curve; a physician needs to complete between 36 and 60 procedures, to achieve clinical proficiency. Marion Surgical K181 is a virtual reality surgical simulator, which emulates the PCNL procedures without compromising the well-being of patients. The simulator uses a VR headset to place a user in a realistic and immersive operating theater, and haptic force-feedback robots to render physical interactions between surgical tools and the virtual patient. The simulator has two modules for two different aspects of PCNL kidney stone removal procedure: kidney access module where the user must insert a needle into the kidney of the patient, and a kidney stone removal module where the user removes the individual stones from the organ. In this paper, we present user trials to validate the face and construct validity of the simulator. The results, based on the data gathered from 4 groups of users independently, indicate that Marion's surgical simulator is a useful tool for teaching and practicing PCNL procedures. The kidney stone removal module of the simulator has proven construct validity by identifying the skill level of different users based on their tool path. We plan to continue evaluating the simulator with a larger sample of users to reinforce our findings.

Abstract This study aimed to characterize the dynamics of suppressor of cytokine signaling (SOCS1) expression in a rat model of lipopolysaccharide-induced periodontitis. Wistar rats in the experimental groups were injected three times/week with LPS from Escherichia coli on the palatal aspect of the first molars, and control animals were injected with vehicle (phosphate-buffered saline). Animals were sacrificed 7, 15, and 30 days after the first injection to analyze inflammation (stereometric analysis), bone loss (macroscopic analysis), gene expression (qRT-PCR), and protein expression/activation (Western blotting). The severity of inflammation and bone loss associated with LPS-induced periodontitis increased from day 7 to day 15, and it was sustained through day 30. Significant (p < 0.05) increases in SOCS1, RANKL, OPG, and IFN-γ gene expression were observed in the experimental group versus the control group at day 15. SOCS1 protein expression and STAT1 and NF-κB activation were increased throughout the 30-day experimental period. Gingival tissues affected by experimental periodontitis express SOCS1, indicating that this protein may potentially downregulate signaling events involved in inflammatory reactions and bone loss and thus may play a relevant role in the development and progression of periodontal disease.

Chronic periodontitis (CP) is an infectious inflammatory disease that affects tooth-supporting structures and in which dental plaque bacteria, immune mechanisms and genetic predisposition play important roles. Interleukin 4 (IL-4) is a key anti-inflammatory cytokine with relevant action in imbalances in inflamed periodontal tissue. Individuals carrying the TCI/CCI genotype (S-haplotype) of the IL-4 gene are 5 times more susceptible to CP, whereas the CTI/TTD genotype (P-haplotype) confers protection against CP. Compared with the S-haplotype, subjects with the P-haplotype produce higher levels of the IL-4 protein after non-surgical periodontal therapy. The present in vitro study aimed to investigate the functionality of IL-4 haplotypes in immune cells to obtain insight into the influence of these genetic variations in regulating immune responses to CP-associated bacteria. Peripheral blood was collected from 6 subjects carrying each haplotype, and their immune cells were challenged with periodontopathogens to compare responses of the different haplotypes with regard to gene expression, protein secretion and the immunophenotype of T helper responses. We found higher IL-4 mRNA and protein levels in the P-haplotype, which also presented higher levels of anti-inflammatory cytokines. In contrast, cells from S-haplotype subjects responded with higher levels of pro-inflammatory cytokines. S-haplotype individuals exhibited significantly greater polarization toward the Th1 phenotype, whereas the P-haplotype was associated with an attenuated response to periodontopathogens, with suggestive skewing toward Th2/M2 phenotypes. In conclusion, IL-4 genetic variations associated with susceptibility to or protection against chronic periodontitis are directly associated with influencing the response of immune cells to periodontopathogens.

Objective: The aim of this study was to evaluate a possible synergism between AGE-RAGE and TLR4 signaling and the role of p38 MAPK and NF-kB signaling pathways on the modulation of the expression of inflammatory cytokines and proliferation of cells from the innate and adaptive immune response. Material and Methods: T lymphocyte (JM) and monocyte (U937) cell lines were stimulated with LPS and AGE-BSA independently and associated, both in the presence and absence of p38 MAPK and NF-kB inhibitors. Proliferation was assessed by direct counting and viability was assessed by a biochemical assay of mitochondrial function. Cytokine gene expression for RAGe, CCL3, CCR5, IL-6 and TNF-α was studied by RT-PCR and RT-qPCR. Results: RAGE mRNA expression was detected in both cell lines. LPS and AGE-BSA did not influence cell proliferation and viability of either cell line up to 72 hours. LPS and LPS associated with AGE induced expression of IL-6 and TNF-α in monocytes and T cells, respectively. Conclusions: There is no synergistic effect between RAGE and TLR signaling on the expression of IL-6, TNF-α , RAGE, CCR5 and CCL3 by monocytes and lymphocytes. Activation of RAGE associated or not with TLR signaling also had no effect on cell proliferation and survival of these cell types.

Abstract Several calcium silicate-based biomaterials have been developed in recent years, in addition to Mineral Trioxide Aggregate (MTA). The aim of this study was to evaluate the cytotoxicity, genotoxicity and apoptosis/necrosis in human osteoblast cells (SAOS-2) of pure calcium silicate-based cements (CSC) and modified formulations: modified calcium silicate-based cements (CSCM) and three resin-based calcium silicate cements (CSCR1) (CSCR 2) (CSCR3). The following tests were performed after 24 hours of cement extract exposure: methyl-thiazolyl tetrazolium (MTT), apoptosis/necrosis assay and comet assay. The negative control (CT-) was performed with untreated cells, and the positive control (CT+) used hydrogen peroxide. The data for MTT and apoptosis were submitted to analysis of variance and Bonferroni’s posttest (p < 0.05), and the data for the comet assay analysis, to the Kruskal-Wallis and Dunn tests (p < 0.05). The MTT test showed no significant difference among the materials in 2 mg/mL and 10 mg/mL concentrations. CSCR3 showed lower cell viability at 10 mg/mL. Only CSC showed lower cell viability at 50 mg/mL. CSCR1, CSCR2 and CSCR3 showed a higher percentage of initial apoptosis than the control in the apoptosis test, after 24 hours exposure. The same cements showed no genotoxicity in the concentration of 2 mg/mL, with the comet assay. CSC and CSCR2 were also not genotoxic at 10 mg/mL. All experimental materials showed viability with MTT. CSC and CSCR2 presented a better response to apoptosis and genotoxicity evaluation in the 10 mg/mL concentration, and demonstrated a considerable potential for use as reparative materials.

Recently, new treatment approaches have been developed to target the host component of periodontal disease. This review aims at providing updated information on host-modulating therapies, focusing on treatment strategies for inhibiting signal transduction pathways involved in inflammation. Pharmacological inhibitors of MAPK, NFκB and JAK/STAT pathways are being developed to manage rheumatoid arthritis, periodontal disease and other inflammatory diseases. Through these agents, inflammatory mediators can be inhibited at cell signaling level, interfering on transcription factors activation and inflammatory gene expression. Although these drugs offer great potential to modulate host response, their main limitations are lack of specificity and developments of side effects. After overcoming these limitations, adjunctive host modulating drugs will provide new therapeutic strategies for periodontal treatment.

The patient's diet has been considered an important etiological factor of dentin hypersensitivity. The frequent ingestion of acidic substances can promote the loss of dental structure or remove the smear layer. The purpose of this study was to evaluate the degree of smear layer removal and dentinal tubules exposure by different natural orange juices. Extracted human teeth were submitted to manual scaling in order to develop the smear layer. Seventy dentin samples were obtained and distributed into the following groups: Control, lime orange, lime, valência orange, navel orange, mandarin, and tangerine. Each group included 2 methods of application: Topical and topical + friction. After preparation for SEM analysis, photomicrographs were assessed by a blind calibrated examiner using an index system. The Kruskal-Wallis test indicated a significant influence of the orange juices on smear layer removal. Significant difference was observed between navel orange, valência orange, mandarin and the control group (p < 0.05). These orange juices resulted in greater removal of the smear layer and greater opening of dentinal tubules. The comparison between the application methods for each group using the Mann-Whitney test showed that friction increased smear layer removal significantly only for lime orange and lime. The data suggest that certain natural orange juices are more effective in terms of smear layer removal and dentinal tubules exposure than others.

The purpose of this clinical study was to investigate if periodontal disease and rheumatoid arthritis (RA) are associated. The study included 39 RA patients (test group) and 22 age- and gender-matched healthy individuals (control group). Questionnaires on general and oral health were applied and a complete periodontal exam, including visible plaque, marginal bleeding, attachment loss (AL) and number of teeth present, was also performed by a single calibrated examiner. Diabetes mellitus patients and smokers were excluded. RA patients had fewer teeth, higher prevalence of sites presenting dental plaque and a higher frequency of sites with advanced attachment loss. Although the prevalence of dental plaque was higher in the test group (Chi-square test, p = 0.0006), the percentage of sites showing gingival bleeding was not different (Fisher’s exact test, p > 0.05). Based on our results, we suggest that there is an association between periodontal disease and RA.

Several antiseptic substances have been used as adjuncts to routine mechanical procedures of oral hygiene, based on their antimicrobial effects. The objective of this study was to assess in vitro the antimicrobial efficiency of a mouthwash containing Triclosan/Gantrez and sodium bicarbonate in comparison to both positive and negative controls. Standard strain samples of Escherichia coli, Pseudomonas aeruginosa, Actinomyces viscosus and Bacillus subtilis were used. Samples of Streptococcus mutans and Gram-negative bacilli were collected from 20 volunteers (10 with a clinically healthy periodontium and 10 presenting biofilm-associated gingivitis). Evaluation of the antimicrobial activity was performed by determining the minimal inhibitory concentration (MIC). The results indicated that the test solution inhibited the growth of both Gram-negative and Gram-positive microorganisms from the volunteers’ saliva as well as that of the standard strains at the MIC dilution of 1:20, whereas the MIC dilution of 0.12% chlorhexidine against the same bacteria was 1:80. Thus, even though the tested mouthrinse solution presented an in-vitro antimicrobial activity superior to that of a placebo, it was inferior to that of chlorhexidine.Diversas substâncias antisépticas têm sido utilizadas como adjuntos aos procedimentos mecânicos rotineiros de higiene oral, com base em seus efeitos antimicrobianos. O objetivo deste estudo foi avaliar, in vitro, a eficiência antimicrobiana de um enxaguatório bucal contendo Triclosan/Gantrez e bicarbonato de sódio, em comparação a controles positivos e negativos. Linhagens padrão de Escherichia coli, Pseudomonas aeruginosa, Actinomyces viscosus e Bacillus subtilis foram utilizadas. Amostras de Streptococcus mutans e Bacilos Gram-negativos foram coletadas de 20 voluntários (10 com um periodonto clinicamente saudável e 10 com gengivite associada à presença de biofilme). A avaliação da atividade antimicrobiana foi realizada pela determinação da Concentração Inibitória Mínima (CIM). Os resultados mostraram que a solução teste inibiu o crescimento de microrganismos Gram-negativos e Gram-positivos da saliva dos voluntários, bem como das linhagens padrão na CIM de 1:20, enquanto que a CIM da diluição de clorexidina 0.12% contra as mesmas bactérias foi de 1:80. Desta forma, apesar de o enxaguatório bucal testado apresentar atividade antimicrobiana in vitro superior à do placebo, esta foi inferior à da Clorexidina.

The aim of the present study was to evaluate the periodontal conditions of anterior teeth that presented pathologic migration in patients with chronic periodontitis and to compare periodontal destruction in migrated versus non-migrated teeth. The sample included 32 patients of both sexes (mean age: 46.0 ± 11.6 years) diagnosed with generalized chronic periodontitis and selected on the basis of the presence of pathologic migration in one or more anterior teeth. This migration was classified according to the following categories: facial flaring, diastema, proximal tilting, rotation or extrusion. The periodontal parameters recorded were clinical attachment loss (CAL) and percentage of radiographic bone loss (BL). Mean CAL of 5.50 ± 2.20 mm and mean BL of 41.90 ± 15.40% were found in 115 teeth assessed. The most frequent type of migration was facial flaring (34.80%), followed by diastema (27.00%). Extrusion was hardly observed in the sample (4.30%). However, greater severity of BL and CAL were observed in teeth with this type of migration (59.44% and 8.42 mm, respectively), and in teeth with facial flaring (45.17% of BL and 6.07 mm of CAL). Kruskal-Wallis test indicated that BL presented by teeth with extrusion or facial flaring was greater than that observed in rotated or tilted teeth (p < 0.05), while there was no difference between groups regarding CAL (p = 0.11). It was observed that anterior teeth with pathologic migration presented greater CAL and BL (5.1 mm and 40%) than non-migrated teeth (4.1 and 31%). The study indicated that the most prevalent kind of pathologic migration is facial flaring, which was associated to higher level of bone loss.O objetivo deste estudo foi avaliar as condições clínicas periodontais de dentes anteriores com migração patológica (MDP) em pacientes com periodontite crônica generalizada e comparar a severidade de destruição periodontal entre dentes migrados e não-migrados. Foram selecionados 32 pacientes, de ambos os sexos, apresentando média de idade de 46,0 anos (± 11,6), com perda clínica de inserção em dentes anteriores e presença de algum tipo de MDP, a saber: vestibularização, diastema, inclinação proximal, giroversão ou extrusão. Os parâmetros avaliados foram a perda clínica de inserção (PIC) e o percentual de perda óssea radiográfica (PO). Os resultados mostraram, em média, uma PIC de 5,50 mm (± 2,20 mm) e uma PO de 41,90% (± 15,40%) do comprimento radicular, em 115 dentes selecionados. Os tipos mais freqüentes de migração foram vestibularização (34,80%) e presença de diastemas (27,00%). A extrusão foi a menos freqüente (4,30%). Maiores valores de PO e PIC foram registrados nos dentes com extrusão (59,44% e 8,42 mm) e vestibularização (45,17% e 6,07 mm). Esses valores de PO foram superiores aos observados nos dentes com giroversão ou inclinação proximal (p < 0,05 - Kruskal-Wallis). A PIC não apresentou diferenças significativas entre os diferentes tipos de migração (p = 0,11). Constatou-se que os dentes anteriores com MDP apresentaram maior PIC e PO (5,1 mm e 40%) quando comparados aos não-migrados (4,1 mm e 31%). Pôde-se concluir também que o tipo de MDP mais prevalente foi a vestibularização, que esteve relacionada a maiores níveis de perda óssea.

Dentin hypersensitivity is a common painful condition observed in clinics. Dietary habits have been much associated with its development and persistence during and following periodontal treatment. The aim of this in vitro study was to evaluate the influence of vinegars on the removal of smear layer and exposure of dentinal tubules. Extracted human teeth were submitted to manual scaling with Gracey curettes in order to remove the cementum as well as to form a smear layer. Dentin samples with 3 mm² were obtained and distributed into six experimental groups: one control and five types of vinegars (alcohol, apple, rice, white wine and balsamic). Each group included two methods of vinegar application: topical and friction. After routine preparation for SEM analysis, photomicrographs were assessed by a calibrated and blind examiner using an appropriate index system. Kruskal-Wallis test indicated a significant influence of vinegars on smear layer removal. There was a statistically significant difference between groups treated with apple, white and rice vinegars and the control group (p < 0.05). Nevertheless, Mann-Whitney test indicated that removal of smear layer did not vary with the method of application (topical versus friction) for any of the tested substances. We can conclude that the contact of vinegar may remove smear layer and expose dentinal tubules, regardless of the type of application. However, balsamic vinegar was associated with less removal of smear layer after both methods of application.A hipersensibilidade dentinária cervical é uma condição dolorosa muito comum nos consultórios. A dieta tem sido bastante associada ao seu aparecimento, assim como a sua persistência após o tratamento periodontal. o objetivo deste trabalho foi avaliar in vitro a influência dos vinagres na remoção de "smear layer" e exposição dos túbulos dentinários. Dentes de humanos foram instrumentados com curetas Gracey para a remoção do cemento e formação de "smear layer". Foram obtidas amostras de dentina com 3 mm², divididas entre o grupo controle (água destilada) e cinco grupos de vinagre: branco, maçã, arroz, vinho branco e balsâmico. Cada grupo incluiu duas formas de aplicação da substância, tópica ou por fricção. Após o preparo para observação em MEV (microscopia eletrônica de varredura), as fotomicrografias foram avaliadas por um examinador previamente calibrado utilizando um índice apropriado. O teste de Kruskal-Wallis indicou influência significativa dos vinagres na remoção de "smear layer". Foi constatada uma diferença estatística significante entre os grupos maçã, branco e arroz e o grupo controle (p < 0,05). O teste de Mann-Whitney, porém, indicou que a remoção de "smear layer" não variou para nenhuma das substâncias segundo a forma de aplicação. Conclui-se que os vinagres podem remover "smear layer" da superfície radicular e expor túbulos dentinários, não sendo influenciados pelo tipo de aplicação. Dentre os tipos de vinagres testados, o balsâmico esteve associado a menor remoção de "smear layer" após ambas as formas de aplicação.

Tetracycline-based matrix metalloproteinase- (MMP-) inhibitors are currently approved for two inflammatory diseases, periodontitis and rosacea. The current study addresses the therapeutic potential of a novel pleiotropic MMP-inhibitor not based on an antibiotic. To induce experimental periodontitis, endotoxin (LPS) was repeatedly injected into the gingiva of rats on one side of the maxilla; the contralateral (control) side received saline injections. Two groups of rats were treated by daily oral intubation with a chemically modified curcumin, CMC 2.24, for two weeks; the control groups received vehicle alone. After sacrifice, gingiva, blood, and maxilla were collected, the jaws were defleshed, and periodontal (alveolar) bone loss was quantified morphometrically and by μ-CT scan. The gingivae were pooled per experimental group, extracted, and analyzed for MMPs (gelatin zymography; western blot) and for cytokines (e.g., IL-1β; ELISA); serum and plasma samples were analyzed for cytokines and MMP-8. The LPS-induced pathologically excessive bone loss was reduced to normal levels based on either morphometric (P=0.003) or μ-CT (P=0.008) analysis. A similar response was seen for MMPs and cytokines in the gingiva and blood. This initial study, on a novel triketonic zinc-binding CMC, indicates potential efficacy on inflammatory mediators and alveolar bone loss in experimental periodontitis and warrants future therapeutic and pharmacokinetic investigations.

The present study aimed to evaluate in vitro whether biomechanical loading modulates proinflammatory and bone remodeling mediators production by periodontal ligament (PDL) cells in the presence of bacterial challenge. Cells were seeded on BioFlex culture plates and exposed to Fusobacterium nucleatum ATCC 25586 and/or cyclic tensile strain (CTS) of low (CTSL) and high (CTSH) magnitudes for 1 and 3 days. Synthesis of cyclooxygenase-2 (COX2) and prostaglandin E2 (PGE2) was evaluated by ELISA. Gene expression and protein secretion of osteoprotegerin (OPG) and receptor activator of nuclear factor kappa-B ligand (RANKL) were evaluated by quantitative RT-PCR and ELISA, respectively. F. nucleatum increased the production of COX2 and PGE2, which was further increased by CTS. F. nucleatum-induced increase of PGE2 synthesis was significantly (P

SOCS3 is an inducible endogenous negative regulator of JAK/STAT pathway, which is relevant in inflammatory conditions. We used a model of LPS-induced periodontal disease in rats to correlate SOCS3 expression with the inflammatory status. In vitro we used a murine macrophage cell line to assess the physical interaction between SOCS3 and STAT3 by coimmunoprecipitation. 30 ug of LPS from Escherichia coli were injected in the gingival tissues on the palatal aspect of first molars of the animals 3x/week for up to 4 weeks. Control animals were injected with the vehicle (PBS). The rats were sacrificed at 7, 15, and 30 days. Inflammation and gene expression were assessed by stereometric analysis, immunohistochemistry, RT-qPCR, and western blot. LPS injections increased inflammation, paralleled by an upregulation of SOCS3, of the proinflammatory cytokines IL-1β, IL-6, and TNF-α and increased phosphorylation of STAT3 and p38 MAPK. SOCS3 expression accompanied the severity of inflammation and the expression of proinflammatory cytokines, as well as the activation status of STAT3 and p38 MAPK. LPS stimulation in a macrophage cell line in vitro induced transient STAT3 activation, which was inversely correlated with a dynamic physical interaction with SOCS3, suggesting that this may be a mechanism for SOCS3 regulatory function.

Suppressor of cytokine signaling 3 (SOCS3) is an inducible endogenous negative regulator of signal transduction and activator of transcription 3 (STAT3). Epigenetic silencing of SOCS3 has been shown in head and neck squamous cell carcinoma (HNSCC), which is associated with increased activation of STAT3. There is scarce information on the functional role of the reduction of SOCS3 expression and no information on altered subcellular localization of SOCS3 in HNSCC.We assessed endogenous SOCS3 expression in different HNSCC cell lines by RT-qPCR and western blot. Immunofluorescence and western blot were used to study the subcellular localization of endogenous SOCS3 induced by IL-6. Overexpression of SOCS3 by CMV-driven plasmids and siRNA-mediated inhibition of endogenous SOCS3 were used to verify the role of SOCS3 on tumor cell proliferation, viability, invasion and migration in vitro. In vivo relevance of SOCS3 expression in HNSCC was studied by quantitative immunohistochemistry of commercially-available tissue microarrays. Endogenous expression of SOCS3 was heterogeneous in four HNSCC cell lines and surprisingly preserved in most of these cell lines. Subcellular localization of endogenous SOCS3 in the HNSCC cell lines was predominantly nuclear as opposed to cytoplasmic in non-neoplasic epithelial cells. Overexpression of SOCS3 produced a relative increase of the protein in the cytoplasmic compartment and significantly inhibited proliferation, migration and invasion, whereas inhibition of endogenous nuclear SOCS3 did not affect these events. Analysis of tissue microarrays indicated that loss of SOCS3 is an early event in HNSCC and was correlated with tumor size and histological grade of dysplasia, but a considerable proportion of cases presented detectable expression of SOCS3.Our data support a role for SOCS3 as a tumor suppressor gene in HNSCC with relevance on proliferation and invasion processes and suggests that abnormal subcellular localization impairs SOCS3 function in HNSCC cells.

The 5-year survival rate for oral cavity cancer is poorer than for breast, colon or prostate cancer, and has improved only slightly in the last three decades. Hence, new therapeutic strategies are urgently needed. Here we demonstrate by tissue micro array analysis for the first time that RNA-binding protein La is significantly overexpressed in oral squamous cell carcinoma (SCC). Within this study we therefore addressed the question whether siRNA-mediated depletion of the La protein may interfere with known tumor-promoting characteristics of head and neck SCC cells. Our studies demonstrate that the La protein promotes cell proliferation, migration and invasion of lymph node-metastasized hypopharyngeal SCC cells. We also reveal that La is required for the expression of β-catenin as well as matrix metalloproteinase type 2 (MMP-2) within these cells. Taken together these data suggest a so far unknown function of the RNA-binding protein La in promoting tumor progression of head and neck SCC.

Mechanical instrumentation of the root surface causes the formation of a smear layer, which is a physical barrier that can affect periodontal regeneration. Although different procedures have been proposed to remove the smear layer, there is no information concerning how long the smear layer persists on root surfaces after instrumentation in vivo. This study assessed the presence of the smear layer on root surfaces over a 28-day period after subgingival instrumentation with hand instruments. Fifty human teeth that were referred for extraction because of advanced periodontal disease were scaled and root planed (SRP) by a single experienced operator. Ten teeth were randomly assigned to be extracted 7, 14, 21, and 28 days after SRP. Another 10 teeth were extracted immediately after instrumentation (Day 0, control group). The subgingival area of the instrumented roots was evaluated with scanning electron microscopy. Representative photomicrographs were assessed by a blinded and calibrated examiner according to a scoring system. A rapid and significant (p < 0.05, Z test) initial reduction in the amount of smear layer was observed at 7 days, and a further significant (p < 0.05) decrease was observed 28 days after SRP. Interestingly, even 28 days after SRP, the smear layer was still present on root surfaces. This study showed that the physiological elimination of the smear layer occurred in a biphasic manner: a rapid initial reduction was observed 7 days after instrumentation, which was followed by a slow process leading to a significant decrease 28 days after instrumentation.