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Metabolomics uses several analytical tools to identify the chemical diversity of metabolites present in organisms. These metabolites are low molecular weight molecules (<1500 Da) classified as a final or intermediary product of metabolic processes. The application of this omics technology has become prominent in inferring physiological conditions through reporting on the phenotypic state; therefore, the introduction of metabolomics into clinical studies has been growing in recent years due to its efficiency in discriminating pathophysiological states. Regarding endocrine diseases, there is a great interest in verifying comprehensive and individualized physiological scenarios, in particular for growth hormone deficiency (GHD). The current GHD diagnostic tests are laborious and invasive and there is no exam with ideal reproducibility and sensitivity for diagnosis neither standard GH cut-off point. Therefore, this review was focussed on articles that applied metabolomics in the search for new biomarkers for GHD. The present work shows that the applications of metabolomics in GHD are still limited, since the little complementarily of analytical techniques, a low number of samples, GHD combined to other deficiencies, and idiopathic diagnosis shows a lack of progress. The results of the research are relevant and similar; however, their results do not provide an application for clinical practice due to the lack of multidisciplinary actions that would be needed to mediate the translation of the knowledge produced in the laboratory, if transferred to the medical setting.

This study aimed to assess the cytotoxicity of commercially available adhesive strategies-etch-and-rinse (Adper ™ Scotchbond ™ 1 XT, 3M ESPE, St. Paul, MN, USA, SB1), self-etch (Clearfil ™ SE Bond 2, Kuraray Noritake Dental Inc., Tokyo, Japan, CSE), and universal (Scotchbond ™ Universal, 3M Deutschland GmbH, Neuss, Germany, SBU). MDPC-23 cells were exposed to adhesives extracts in different concentrations and exposure times. To access cell metabolic activity, viability, types of cell death, and cell cycle, the MTT assay, SRB assay, double labeling with annexin V and propidium iodide, and labeling with propidium iodide/RNAse were performed, respectively. Cultures were stained with May-Grünwald Giemsa for qualitative cytotoxicity assessment. The SB1, CSE, and SBU extracts determined a significant reduction in cell metabolism and viability. This reduction was higher for prolonged exposures, even for less concentrated extracts. CSE extracts significantly reduced the cell's metabolic activity at higher concentrations (50% and 100%) from 2 h of exposure. After 24 and 96 h, a metabolic activity reduction was verified for all adhesives, even at lower concentrations. These changes were dependent on the adhesive, its concentration, and the incubation time. Regarding cell viability, SBU extracts were the least cytotoxic, and CSE was significantly more cytotoxic than SB1 and SBU. The adhesives determined a reduction in viable cells and an increase in apoptotic, late apoptosis/necrosis, and necrotic cells. Moreover, on cultures exposed to SB1 and CSE extracts, a decrease in the cells in S and G2/M phases and an increase in the cells in G0/G1 phase was observed. Exposure to SBU led to an increase of cells in the S phase. In general, all adhesives determined cytotoxicity. CSE extracts were the most cytotoxic and were classified as having a higher degree of reactivity, leading to more significant inhibition of cell growth and destruction of the cell's layers.

The aim of this study was to evaluate and compare the sealing ability of two calcium silicate-based sealers (TotalFill BC RRM Fast Set Putty and White ProRoot MTA) when used as apical plugs in immature teeth through nuclear medicine. Single-rooted extracted teeth (n = 34) had their crowns and root tip sectioned to obtain 14 mm long root segments to simulate an in vitro apexification model. Were created two experimental groups, namely MTA (n = 12) and BC (n = 12), and two control groups, PG (positive group, n = 5) and NG (negative group, n = 5). On the 4th day after placing the respective apical plug, the apical portions of the teeth were submerged in a solution of sodium pertechnetate ( 99m TcNaO 4 ) for 3 h. Statistical analysis showed a significant difference between the MTA group and the controls ( p < 0.05). The BC group had a significant difference regarding the negative control ( p < 0.001) but showed no statistical significance regarding the positive control ( p = 0.168). There was a statistically significant difference ( p = 0.009) between the BC group (7335.8 ± 2755.5) and the MTA group (4059.1 ± 1231.1), where the last showed less infiltration. Within the limitations of this study, White ProRoot MTA had a significantly better sealing ability than TotalFill BC RRM Fast Set Putty.

Hypericin (Hy) is a hydrophobic photosensitizer used in photodynamic therapy for cancer therapeutic. In this study, Hy-loaded oil-in-water (O/W) nanoemulsions (NEs) were produced by the ultrasonication method combing different biocompatible oils and surfactants to enhance Hy aqueous solubility and bioavailability. Experimental parameters were optimized by the characterization of droplet size, zeta potential, and physicochemical properties. In vitro studies based on the release profile, cytotoxicity, cell morphology, and Hy intracellular accumulation were assayed. Hy at 100 mg L -1 was incorporated into the low viscosity (~0.005 Pa s) NEs with spherical droplets averaging 20-40 nm in size and polydispersity index <0.02. Hy release from the NE was significantly higher (4-fold) than its suspension (p < 0.001). The NEs demonstrated good physical stability during storage at 5 °C for at least six months. The Hy-loaded NEs exhibited an IC50 value 6-fold lower than Hy suspension during PDT against breast cancer cell lines (MCF-7). Cell microscopy imaging confirmed the increased cytotoxic effects of Hy-loaded NEs, showing damaged and apoptotic cells. Confocal laser scanning microscopy evidenced greater Hy delivery through NE into MCF-7 cells followed by improved intracellular ROS generation. Our results suggest that the Hy-loaded NEs can improve hypericin efficacy and assist Hy-PDT's preclinical development as a cancer treatment., (Copyright © 2021 Elsevier B.V. All rights reserved.)

To increase milk production, antibiotics are administered to animals to provide weight gain and to prevent or treat diseases. The inappropriate use of these substances can lead to antibiotic resistance and allergic reactions and toxic effects to milk consumers. We describe the development of a simple, fast, portable, and low-cost microfluidic paper-based analytical device (μPAD) to quantify sulfonamides in milk using the inhibition of the colorimetric reaction between carbonic anhydrase (CA) and 4-nitrophenyl acetate. The main advantages presented by the μPAD include reproducible batch production, simple application, and precise analysis without previous treatment. The µPAD displayed good linearity (R 2  ≥ 0.986) in a wide range of sulfonamides in milk (2.5 to 40.0 µmol L -1 ), being selective for the drugs even in a highly complex matrix. We expect that this device allows in situ monitoring of milk quality, reducing the prejudicial conditions associated with high concentrations of sulfonamides in milk., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Biomaterials contact directly or indirectly with the human tissues, making it important to evaluate its cytotoxicity. This evaluation can be performed by several methods, but a high discrepancy exists between the approaches used, compromising the reproducibility and the comparison among the obtained results. In this paper, we propose a protocol to evaluate biomaterials cytotoxicity using soluble extracts, which we use for dental biomaterials. The extracts preparation is detailed, from pellets production to its extraction in a culture medium. The biomaterials cytotoxicity evaluation is based on metabolic activity using the MTT assay, cell viability using the Sulphorhodamine B (SBR) assay, cell death profile by flow cytometry, and cell morphology using May-Grünwald Giemsa. Additional to cytotoxicity evaluation, a protocol to evaluate cell function is described based on the expression of specific markers assessed by immunocytochemistry and PCR. This protocol provides a comprehensive guide for biomaterials cytotoxicity and cellular effects evaluation, using the extracts methodology, in a reproducible and robust manner.

The occlusal matrix technique appears as an alternative to the conventional restorative technique. The aim of this study is to present treatment options for the decayed occlusal surface of posterior teeth by means of an occlusal matrix made with different materials, for an accurate reproduction of the tooth surface. This paper presents five clinical cases using five different materials, such as Bite-Perf, flowable composite, self-curing acrylic resin, silicone and thermoforming foil. Before light-curing, the last layer of composite resin, the matrix, is repositioned and firmly pressed so the replication of the original occlusal anatomy can be achieved. When using an occlusal matrix, minimal or no finishing and polishing procedures are needed and the offered aesthetics are clearly an advantage. Moreover, this technique makes up for the additional chairside work time. The occlusal matrix technique is demonstrated to be effective for direct composite resin restorations in posterior teeth, allowing a reliable and predictable reproduction of the occlusal anatomy and clinical adjustment. For this purpose, a clear material is preferable since it allows light transmission., (© 2021. The Author(s), under exclusive licence to the British Dental Association.)

Cancer cells have differential metabolic dependencies compared to their nonmalignant counterparts. However, few metabolism-targeting compounds have been successful in clinical trials. Here, we investigated the metabolic vulnerabilities of triple-negative breast cancer (TNBC), particularly those metabolic perturbations that increased mitochondrial apoptotic priming and sensitivity to BH3 mimetics (drugs that antagonize antiapoptotic proteins). We used high-throughput dynamic BH3 profiling (HT-DBP) to screen a library of metabolism-perturbing small molecules, which revealed inhibitors of the enzyme nicotinamide phosphoribosyltransferase (NAMPT) as top candidates. In some TNBC cells but not in nonmalignant cells, NAMPT inhibitors increased overall apoptotic priming and induced dependencies on specific antiapoptotic BCL-2 family members. Treatment of TNBC cells with NAMPT inhibitors sensitized them to subsequent treatment with BH3 mimetics. The combination of a NAMPT inhibitor (FK866) and an MCL-1 antagonist (S63845) reduced tumor growth in a TNBC patient-derived xenograft model in vivo. We found that NAMPT inhibition reduced NAD + concentrations below a critical threshold that resulted in depletion of adenine, which was the metabolic trigger that primed TNBC cells for apoptosis. These findings demonstrate a close interaction between metabolic and mitochondrial apoptotic signaling pathways and reveal that exploitation of a tumor-specific metabolic vulnerability can sensitize some TNBC to BH3 mimetics., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

(1) Background: Alveolar bone defects or decreased alveolar bone height and width may have different causes, such as cleft palate. Regenerative procedures in oro-dental defects are challenging due to anatomical factors and the distinct cell populations involved. The iliac crest bone graft remains the gold-standard for cleft palate closure. However, tissue regeneration approaches have been employed and their outcome reviewed, but no conclusions have been made about which one is the gold-standard. (2) Methods: this umbrella review aims to critically appraise the effectiveness of the current approaches in bone defects regeneration in non-syndromic patients with cleft palate. A search was performed in PubMed, Cochrane Library, Scopus, Web of Science and EMBASE databases. (3) Results: Systematic reviews of randomized and non-randomized controlled trials with or without meta-analysis were included. Nine articles were included in the qualitative analysis and five in the quantitative one. The included studies quality was evaluated with AMSTAR2. (4) Conclusions: The use of new regenerative strategies, such as bone morphogenic protein 2, appears to provide similar results regarding bone volume, filling, and height to the standard technique with the iliac crest bone graft.

Coronavirus disease 2019 (COVID-19), a viral disease declared a pandemic by the World Health Organization (WHO) in March 2020, has posed great changes to many sectors of society across the globe. Its virulence and rapid dissemination have forced the adoption of strict public health measures in most countries, which, collaterally, resulted in economic hardship. This article is the first in a series of 3 that aims to contextualise the clinical impact of COVID-19 for the dental profession. It presents the epidemiological conditions of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), namely, its modes of transmission, incubation, and transmissibility period, signs and symptoms, immunity, immunological tests, and risk management in dental care. Individuals in dental care settings are exposed to 3 potential sources of contamination with COVID-19: close interpersonal contacts (<1 m), contact with saliva, and aerosol-generating dental procedures. Thus, a risk management model is propsoed for the provision of dental care depending on the epidemiological setting, the patient's characteristics, and the type of procedures performed in the office environment. Although herd immunity seems difficult to achieve, a significant number of people has been infected throughout the first 9 months of the pandemic and vaccination has been implemented, which means that there will be a growing number of presumable "immune" individuals that might not require many precautions that differ from those before COVID-19. In conclusion, dental care professionals may manage their risk by following the proposed model, which considers the recommendations by local and international health authorities, thus providing a safe environment for both professionals and patients., Competing Interests: Conflict of interest None disclosed., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Viruses are the causing agents for many relevant diseases, including influenza, Ebola, HIV/AIDS, and COVID-19. Its rapid replication and high transmissibility can lead to serious consequences not only to the individual but also to collective health, causing deep economic impacts. In this scenario, diagnosis tools are of significant importance, allowing the rapid, precise, and low-cost testing of a substantial number of individuals. Currently, PCR-based techniques are the gold standard for the diagnosis of viral diseases. Although these allow the diagnosis of different illnesses with high precision, they still present significant drawbacks. Their main disadvantages include long periods for obtaining results and the need for specialized professionals and equipment, requiring the tests to be performed in research centers. In this scenario, biosensors have been presented as promising alternatives for the rapid, precise, low-cost, and on-site diagnosis of viral diseases. This critical review article describes the advancements achieved in the last five years regarding electrochemical biosensors for the diagnosis of viral infections. First, genosensors and aptasensors for the detection of virus and the diagnosis of viral diseases are presented in detail regarding probe immobilization approaches, detection methods (label-free and sandwich), and amplification strategies. Following, immunosensors are highlighted, including many different construction strategies such as label-free, sandwich, competitive, and lateral-flow assays. Then, biosensors for the detection of viral-diseases-related biomarkers are presented and discussed, as well as point of care systems and their advantages when compared to traditional techniques. Last, the difficulties of commercializing electrochemical devices are critically discussed in conjunction with future trends such as lab-on-a-chip and flexible sensors., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Aim: Evaluate the chemopreventive potential of the extract from P. polymyxa RNC-D. Methods: Concentrations of P. polymyxa RNC-D extract were tested in HepG2/C3A cells to assess their genotoxic (comet assay), mutagenic (micronucleus test) and antigenotoxic potential (comet assay) in vitro . Results: 400 and 40 μg/ml concentrations induced DNA lesions, whereas the 4 μg/ml induced a desmutagenic effect. Complementary tests indicated that the extract minimized the formation of reactive oxygen species induced by methyl methanesulfonate and normalized the loss of membrane potential. The quantification of cytokines indicated that TNF-α was immunostimulated by the extract. However, when administered in conjunction with the methyl methanesulfonate, the extract blocked the TNF-α release. Conclusion: The fermentation broth from P. polymyxa RNC-D showed an antigenotoxic effect, and thus the potential to be used as chemopreventive compound.

Some authors have been proposing the use of cavity disinfectants in order to reduce, or even eliminate, the effect of the microorganisms present in a dental cavity before a restoration is placed. The aim of this study was to evaluate the effect of different cavity disinfectants on bond strength and clinical success of composite and glass ionomer restorations on primary teeth. The research was conducted using Cochrane Library, PubMed/MEDLINE, SCOPUS, and Web of Science for articles published up to February 2021. The search was performed according to the PICO strategy. The evaluation of the methodological quality of each in vitro study was assessed using the CONSORT checklist for reporting in vitro studies on dental materials. Sixteen in vitro studies and one in situ study fulfilled the inclusion criteria and were analyzed. Chlorhexidine was the most studied cavity disinfectant, and its use does not compromise dentin bonding. Sodium hypochlorite is a promising alternative, but more research on its use is required to clearly state that it can safely be used as a cavity disinfectant for primary teeth. Although other disinfectants were studied, there is a low-level evidence attesting their effects on adhesion, therefore their use should be avoided.

The surface smoothness of composite restorations affects not only their esthetic appearance but also other properties. Thus, rough surfaces can lead to staining, plaque accumulation, gingival irritation, recurrent caries, abrasiveness, wear kinetics, and tactile perception by the patient. The aim of this study was to evaluate the influence of irrigation during the finishing and polishing of composite resin restorations. A systematic search of the PubMed, Cochrane Library, EMBASE, Web of Science, and Clinical Trials databases was conducted. Papers published up to 11 February 2021 were considered. The quality of each study was assessed using the modified Consolidated Standards of Reporting Trials checklist for reporting in vitro studies on dental materials. No clinical studies were identified. Six in vitro studies were included, reporting changes in physical and esthetic properties. After performing a methodological quality assessment of the studies, some limitations were identified, the main limitation being the heterogeneous methodology across studies. The evidence resulting from this systematic review did not favor either wet or dry finishing/polishing procedures. There is a clear need for well-designed studies focusing on the comparison of dry/wet finishing/polishing with standard protocols to evaluate the differences among different materials and methods.

Microfluidics is an essential technique used in the development of in vitro models for mimicking complex biological systems. The microchip with microfluidic flows offers the precise control of the microenvironment where the cells can grow and structure inside channels to resemble in vivo conditions allowing a proper cellular response investigation. Hence, this study aimed to develop low-cost, simple microchips to simulate the shear stress effect on the human umbilical vein endothelial cells (HUVEC). Differentially from other biological microfluidic devices described in the literature, we used readily available tools like heat-lamination, toner printer, laser cutter and biocompatible double-sided adhesive tapes to bind different layers of materials together, forming a designed composite with a microchannel. In addition, we screened alternative substrates, including polyester-toner, polyester-vinyl, glass, Permanox ® and polystyrene to compose the microchips for optimizing cell adhesion, then enabling these microdevices when coupled to a syringe pump, the cells can withstand the fluid shear stress range from 1 to 4 dyne cm 2 . The cell viability was monitored by acridine orange/ethidium bromide (AO/EB) staining to detect live and dead cells. As a result, our fabrication processes were cost-effective and straightforward. The materials investigated in the assembling of the microchips exhibited good cell viability and biocompatibility, providing a dynamic microenvironment for cell proliferation. Therefore, we suggest that these microchips could be available everywhere, allowing in vitro assays for daily laboratory experiments and further developing the organ-on-a-chip concept.

Cavity disinfection becomes an important step before a dental restorative procedure. The disinfection can be obtained cleaning the dental cavity with antimicrobial agents before the use of adhesive systems. The aim of this study was to conduct a systematic review on the effect of different cavity disinfectants on restorations' adhesion and clinical success. A search was carried out through the Cochrane Library, PubMed, and Web of Science. In vitro and in situ studies reporting results on dentin bond strength tests, and clinical studies published until August 2020, in English, Spanish and Portuguese were included. The methodological quality assessment of the clinical studies was carried out using the Revised Cochrane risk-of-bias tool. Chlorhexidine could preserve adhesion to dentin. EDTA and ethanol had positive results that should be further confirmed. Given the significant lack of scientific evidence, the use of lasers, fluoridated agents, sodium hypochlorite, or other products as cavity disinfectants should be avoided. Chlorhexidine is a safe option for cavity disinfection with adequate preservation of adhesion to dentin. Moreover, future researches should be focused on the efficacy of these disinfectants against cariogenic bacteria and their best application methods.

(1) Aim: This study aims to analyze the in vitro infiltration of a silicate root canal sealer into dentinal tubules after using different endodontic irrigating solutions. (2) Methods: Twenty-nine teeth with single roots were separated into three groups according to the final irrigation protocol: G1 n = 10) = 17% EDTA (ethylenediaminetetraacetic acid) + 3.0% sodium hypochlorite (NaOCl), G2 ( n = 10) = 17% EDTA + 2.0% chlorhexidine and G3 (Control group, n = 9) = 17% EDTA + saline solution. Root canals were filled using cold lateral compaction technique with MTA Fillapex sealer and gutta-percha. The sealer was labeled with rhodamine B. The teeth were segmented at the middle and third apical sections, which were visualized using 10× confocal laser microscopy to determine the sealer penetration percentage. (3) Results: In the apical section, no statistically significant differences were found between the groups regarding sealer penetration. In the middle section, Group 1 obtained the highest percentage, and Group 2 the lowest ( p = 0.004). Group 1 also presented statistically significant differences in the Control Group ( p = 0.031) and had close sealer penetration values. Meanwhile, the Control Group ( p = 0.023) and Group 2 ( p = 0.029) revealed a significant decrease of sealer penetration between the apical and middle sections. (4) Conclusion: The obtained results support that final irrigation with NaOCl promoted similar sealer penetration in the apical and middle sections. On the other hand, a significant decrease in the sealer penetration of the middle section was observed for the chlorhexidine and saline groups. Compared to other irrigant solutions, NaOCl promotes more uniform sealer penetration, which can correlate with better sealing and, consequently, higher endodontic treatment success.

Angiostrongylus cantonensis is the main cause of human eosinophilic meningitis. Humans are accidental hosts, becoming infected due to ingestion of raw intermediate (snails and slugs) or paratenic hosts. Once ingested, the larvae migrate towards the brain where they die, causing the disease. To develop better mollusk control strategies, it is important to first understand what happens in the snail during infection, therefore our purpose was to characterize proteomic, metabolic and immunologic changes in Biomphalaria glabrata 24 h after infection with A. cantonensis. For this purpose, proteins were extracted from infected and uninfected snails and analyzed through mass spectrometry. Hemolymph was also collected, the number of hemocytes was counted and urea, nitric oxide, calcium, glycogen levels as well as alanine and aspartate aminotransferases activities were assessed. The cephalopodal region and gonad-digestive gland complex were dissected and their glycogen content was measured. After infection with A. cantonensis, we observed an increase of hemocytes and granulocytes as well as an increase in hemoglobin type 2 proteins. Temptin-like protein was also found up-regulated in infected snails. Several proteins with structural function (such as myosin heavy chain - striated muscle - like and protein LOC106059779 with ADAM/reprosolin domain) were also differentially expressed, suggesting loss/damage of internal tissues. Increase in phosphoglycerate mutase indicates an increase in glycolysis, possible to compensate the increase in energetic needs. Consequently, there is a decrease in glycogen reserves, particularly in the gonad - digestive gland complex., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Dental caries is a disease of dental hard tissues, considered the most common non-communicable disease worldwide. Conventional treatments for caries removal are often associated with pain and fear, so different therapeutic approaches have been developed towards more conservative and pleasant treatments. This systematic review aimed to assess the efficacy and patient's acceptance of alternative methods for caries removal compared to conventional methods. The Cochrane Library, Embase, Medline/Pubmed, Web of Science, and Clinical Trials databases were searched. Clinical trials of primary dental caries treated with alternative methods were included. The last search was performed on 5 August 2020. The Population, Intervention, Comparison, and Outcome (PICO) strategy was followed. Thirty-seven clinical trials were included, reporting caries removal using alternative (chemomechanical-Brix 3000, Carie-care, Carisolv and Papacarie, laser-Er:YAG (Erbium-doped Yttrium Aluminium Garnet) and Er,Cr:YSGG (Erbium, Chromium-doped Yttrium, Scandium, Gallium and Garnet), and a system combining air and sono-abrasion-Vector ® System) and conventional methods. Alternative methods tended to prolong treatment time and lessen anesthesia need. All treatments were effective in reducing cariogenic flora, and the restoration's performance did not differ significantly. Chemomechanical solutions seemed to be the best option towards minimally invasive treatments, with good control during application and action and good treatment experiences for patients. Papacarie was demonstrated to be an effective method for caries removal with less pain, and superior acceptance by patients when compared to conventional treatments., Competing Interests: The authors declare no conflict of interest.

The sealers used for root canal treatment should be biocompatible for the peri-radicular tissues, to evaluate the cytotoxic effects of GuttaFlow ® bioseal sealer and to compare them with AH26 ® epoxy resin. Culture media were conditioned with the GuttaFlow ® bioseal and AH26 ® pellets. MDPC-23 odontoblast cell cultures were treated with conditioned medium and serial dilutions. To evaluate the metabolic activity and cellular viability, the MTT and SRB assays were performed. To determine the production of reactive oxygen species, the DHE and DCF-DA probes were used. Cell cycle and cell-death types were assessed by cytometry, and to evaluate the mineralization capacity, the Alizarin Red S coloration was used. Statistical analysis was performed using analysis of variance (ANOVA) when normality was found and Kruskal-Wallis on the opposite case. For the comparison with normality values, the Student t -test was used. Cells exposed to the GuttaFlow ® bioseal conditioned medium maintained high metabolic activities, except at higher concentrations. Likewise, viability was maintained, but a significant decrease was observed after exposure to the highest concentration ( p < 0.001), associated with cell death by late apoptosis and necrosis. When cell cultures were exposed to AH26 ® , metabolic activity was highly compromised, resulting in cell death. An imbalance in the production of peroxides and superoxide anion was observed. GuttaFlow ® bioseal showed higher biocompatibility than AH26 ® .

Cri Du Chat (CDC) syndrome is a rare genetic condition caused by the deletion of genetic material on the small arm (the p arm) of chromosome 5. A high-pitched cry that sounds like that of a cat, dysmorphic characteristics, and cytogenetic methods are often used for diagnosing the syndrome. In this study, we applied GC-MS analysis for determining organic acids in urine from 17 control volunteers without CDC syndrome, and from 16 individuals with the CDC syndrome in order to determine the profile of organic acids and biochemical pathways alterations resulting from this genetic condition. First, performing multivariate data analysis selected the best method for extracting organic acids with greater signal intensities and good reproducibility. After selection, multivariate (PLS-DA) and univariate (Mann-Whitney test) data analysis discriminated the metabolites responsible for separation between groups. Nine organic acid metabolites had values of VIP ≥ 1.0 and p-values ≤ 0.05, with highest intensities in the samples from CDC individuals, indicating the strongest discriminative power (tricarballylic acid, indoleacetic acid, anthranilic acid, 4-hydroxyphenylacetic acid, 4-hydroxybenzoic acid, 4-hydroxyhippuric acid, pantothenic acid, homovanillic acid, and vanillylmandelic acid). These metabolites are involved in several biochemical pathways like in the tyrosine and phenylalanine metabolism, as well as the tryptophan metabolism, which could be associated (i) to some neuropsychiatric alterations commonly observed in CDC individuals, (ii) to exogenous compounds related to transformation products by intestinal microbial, and (iii) to a possible deficiency in enzyme activity due to the syndrome., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Extracellular vesicles (EVs) are a frontier class of circulating biomarkers for the diagnosis and prognosis of different diseases. These lipid structures afford various biomarkers such as the concentrations of the EVs ( C V ) themselves and carried proteins ( C P ). However, simple, high-throughput, and accurate determination of these targets remains a key challenge. Herein, we address the simultaneous monitoring of C V and C P from a single impedance spectrum without using recognizing elements by combining a multidimensional sensor and machine learning models. This multidetermination is essential for diagnostic accuracy because of the heterogeneous composition of EVs and their molecular cargoes both within the tumor itself and among patients. Pencil HB cores acting as electric double-layer capacitors were integrated into a scalable microfluidic device, whereas supervised models provided accurate predictions, even from a small number of training samples. User-friendly measurements were performed with sample-to-answer data processing on a smartphone. This new platform further showed the highest throughput when compared with the techniques described in the literature to quantify EVs biomarkers. Our results shed light on a method with the ability to determine multiple EVs biomarkers in a simple and fast way, providing a promising platform to translate biofluid-based diagnostics into clinical workflows.

Epithelial to Mesenchymal Transition (EMT) is a normal cellular process that is also triggered during cancer progression and metastasis. EMT induces cellular and microenviromental changes, resulting in loss of epithelial features and acquisition of mesenchymal phenotypes. The growth factor TGFβ and the transcription factor SNAIL1 (SNAIL) have been described as inducers of EMT. Here, we carried out an EMT model with non-tumorigenic cell line MCF-10A induced with the TGFβ2 specific isoform of TGF protein family. The model was validated by molecular, morphological and functional experiments and showed correlation with the up-regulation of SNAIL. In order to identify additional regulators of EMT in this non-tumorigenic model, we explored quantitative proteomics, which revealed the Ubiquitin carboxyl-terminal hydrolase 47 (USP47) as one of the top up-regulated proteins. USP47 has a known role in cell growth and genome integrity, but not previously correlated to EMT. After validating USP47 alterations using MRM and antibody-based assays, we demonstrated that the chemical inhibition of USP47 with the inhibitor P5091 reduced expression of EMT markers and reverted morphological changes in MCF-10A cells undergoing EMT. These results support the involvement of USP47 in our EMT model as well as potential applications of deubiquitinases as therapeutic targets for cancer progression management. BIOLOGICAL SIGNIFICANCE: Metastasis is responsible for most cancer-associated mortality. Additionally, metastasis requires special attention, as the cellular transformations make treatment at this stage very difficult or occasionally impossible. Early steps in cancer metastasis involve the ability to detach from the solid tumor mass and invade the surrounding stromal tissues through cohesive migration, or a mesenchymal or amoeboid invasion. One of the first steps for metastatic cascade is denominated epithelial to mesenchymal transition (EMT), which can be triggered by different factors. Here, our efforts were directed to better understand this process and identify new pathways that contributes for acquisition of EMT, mainly focused on post translational modifications related to ubiquitin proteasome system. Our model of EMT induction by TGFβ2 mimics early stage of metastatic cancer in epithelial breast cells and a proteomic study carried out for such model demonstrates that the deubiquitinase enzyme USP47 acts in SNAIL stabilization, one of the most important transcription factors for EMT phenotype acquisition and consequent metastasis. In addition, the inhibiton of USP47 with P5091, reverted the EMT phenotype. Together the knowledge of such processes of cancer progression and regulation can help in designing new strategies for combined therapies for control of cancer in early stages., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Bisphosphonate-associated osteonecrosis of the jaw (BRONJ), a post-surgical non-healing wound condition, is one of the most common side effects in patients treated with nitrogen-containing bisphosphonates. Its physiopathology has been related with suppression of bone turnover, of soft tissue healing and infection. Biphasic calcium phosphates (BCP) are used as a drug delivery vehicle and as a bone substitute in surgical wounds. Due to their capacity to adsorb zoledronate, it was hypothesized these compounds might have a protective effect on the soft tissues in BRONJ wounds. To address this hypothesis, a reproducible in vivo model of BRONJ in Wistar rats was used. This model directly relates chronic bisphosphonate administration with the development of osteonecrosis of the jaw after tooth extraction. BCP granules were placed in the alveolus immediately after tooth extraction in the test group. The animals were evaluated through nuclear medicine, radiology, macroscopic observation, and histologic analysis. Encouragingly, calcium phosphate ceramics were able to limit zoledronate toxicity in vivo and to favor healing, which was evidenced by medical imaging (nuclear medicine and radiology), macroscopically, and through histology. The studied therapeutic option presented itself as a potential solution to prevent the development of maxillary osteonecrosis.

The oxidation of paper by periodate was investigated and systematically characterized by Fourier-transform infrared (FTIR) spectroscopy, scanning electron microscopy, X-ray diffraction, goniometry, and dynamic mechanical analysis. For the first time, in situ FTIR microscopy analysis was performed, yielding chemical images of carbonyl groups on the cellulose fibers. The enhancement of protein immobilization on oxidized paper was quantified by a colorimetric assay with Ponceau dye, demonstrating that 0.5-h oxidation suffices to functionalize the paper-based devices. The oxidized paper was applied as a sensor for protein quantification in urine, a test able to detect levels of proteinuria and even microalbuminuria. The quantification was based on the capture of proteins through covalent bonds formed with the carbonyl groups on the oxidized paper followed by the staining of the region with Ponceau dye. There is a linear dependency between human serum albumin (HSA) concentration and the length of the stained blot from 0.1 to 3 mg mL -1 . This method correlated linearly with a reference method showing a higher sensitivity (0.866 cm mL mg -1 ) than the latter. The limit of quantification was 0.1 mg mL -1 , three times lower than that of the commercial strip. Graphical abstract Paper oxidation with periodate and extensive characterization, including microspectroscopy. The conversion of cellulose hydroxyl groups to aldehyde enhances covalent immobilization of protein on paper for application as analytical device. The oxidized paper determined protein in urine, suitable for proteinuria diagnosis.

The aim of this study was to evaluate the cytotoxic effects of an enzymatic mouthwash and of a chlorhexidine mouthwash on human gingival fibroblasts. The metabolic activity of the fibroblasts exposed to each mouthwash was assessed by the MTT assay and the protein content was assessed by the SRB assay. The flow cytometry was used to evaluate the cell cycle and the types of cell death. The oxidative status was evaluated through the DCF and the DHE probes and the intracellular GSH concentration and the mitochondrial membrane potential through JC-1. The cytotoxicity of both mouthwashes was found to be dependent on the exposure time and on the concentration. However, the cytotoxicity of the enzymatic mouthwash was found to be lower than that of the chlorhexidine mouthwash. A trend towards increased oxidative stress was observed for both mouthwashes. After exposing the fibroblasts to the mouthwashes, a G2/M phase block was observed and cell death occurred predominantly by necrosis. The effects of chlorhexidine on fibroblasts were identified at lower concentrations than those used in clinical practice. Therefore, the use of chlorhexidine as an antiseptic in surgical and postoperative situations should be limited. In order to clarify the clinical significance of the enzymatic mouthwash cytotoxicity new clinical studies will be necessary.

Tooth whitening procedures are increasing; however, side effects can occur, such as damage to pulp cells, by the whitening products. This study aims to assess the cellular effects promoted by a whitening product, namely, the oxidative stress fostered by the active agent hydrogen peroxide, with and without photoactivation. Additionally, if cellular recovery occurred, we intended to determine the time point where cells recover from the tooth whitening induced damage. Human fibroblasts were exposed to hydrogen peroxide, Zoom ® , Zoom ® + irradiation, and irradiation alone. The following analysis was performed: metabolic activity evaluation by the MTT assay; cell viability, mitochondrial membrane potential, peroxides production, superoxide radical production, and reduced glutathione expression by flow cytometry. We determined the IC 50 value for all groups, and a dose-dependent cytotoxic effect was verified. At the times analyzed, hydrogen peroxide groups showed no metabolic activity recovery while a cell recovery was observed after 24 h (Zoom ® ) and 48 h (Zoom ® + irradiation). Cell death was seen in hydrogen peroxide and Zoom ® + irradiation groups, mainly by apoptosis, and the irradiation had a cytotoxic effect per se. This in vitro study supports that whitening products with moderate hydrogen peroxide (HP) concentration have a temporary effect on cells, allowing a cellular recovery.

Background: Indirect restorations using composites with ceramic fillings can be an alternative to ceramic veneering and direct composite restorations for the treatment of posterior teeth. The aim of this study was the evaluation of the mechanical and tribological properties of a ceromer., Materials and Methods: Sixty specimens were produced and divided into two groups: one control group not submitted to thermocycling (n = 20) and one test group submitted to 5000 cycles of thermocycling (n = 40). The studied parameters were microhardness, surface roughness and the coefficient of friction (scratch test)., Results: The ceromer exhibits a reduction of polymerization shrinkage, higher wear, and fracture resistance than the composite resins. The studied ceromer presented good mechanical properties, even after being submitted to thermocycling. Roughness was the property most affected, increasing 25.8%, microhardness decreased by 10.5% and the coefficient of friction increased by 4.2%., Conclusions: In certain situations, ceromers can be an alternative to composite resins and ceramics, providing an aesthetic, conservative and longevity option.

(1) Aim: To perform a systematic review of the literature on the biocompatibility of root canal sealers that encompasses the various types of sealers that are commercially available as well as both in vitro and in vivo evidence. (2) Methods: This systematic review has been registered in PROSPERO (ID 140445) and was carried out according to PRISMA guidelines using the following databases: PubMed, Cochrane Library, ClinicalTrials.gov, Science Direct, and Web of Science Core Collection. Studies published between 2000 and 11 June 2019 that evaluated cytotoxicity (cell viability/proliferation) and biocompatibility (tissue response) of root canal sealers were included. (3) Results: From a total of 1249 studies, 73 in vitro and 21 in vivo studies were included. In general, studies suggest that root canal sealers elicit mild to severe toxic effects and that several factors may influence biocompatibility, e.g., material setting condition and time, material concentration, and type of exposure. Bioactive endodontic sealers seem to exhibit a lower toxic potential in vitro. (4) Conclusions: The available evidence shows that root canal sealers exhibit variable toxic potential at the cellular and tissue level. However, the methodological heterogeneity among studies included in this systematic review and the somewhat conflicting results do not allow a conclusion on which type of sealer presents higher biocompatibility. Further research is crucial to achieve a better understanding of the biological effects of root canal sealers.

Mansonic schistosomiasis is a neglected disease transmitted by Biomphalaria spp. snails. Understanding what happens inside the intermediate host is important to develop more efficient ways of reducing schistosomiasis prevalence. Our purpose was to characterize metabolic and immunological changes in Biomphalaria glabrata 24 h after exposure to Schistosoma mansoni. For this purpose, proteins were extracted from snails' whole tissue with Tris-Urea buffer and digested with tripsin. Mass spectrometry was performed and analyzed with MaxQuant and Perseus software. Also, the hemolymph of five snails 24 h post exposure was collected, and the numbers of hemocytes, levels of urea, uric acid, nitric oxide, calcium, glycogen and alanine and aspartate aminotransferases activities were assessed. Snails were also dissected for measurement of glycogen content in the cephalopodal region and gonoda-digestive gland complex. Globin domain proteins were found to be up-regulated; also the number of circulating hemocytes was significantly higher after 24 h of exposure to the parasite. NO levels were higher 24 h post exposure. Several proteins associated with energy metabolism were found to be up-regulated. Glycogen analysis showed a significant decrease in the gonad-digestive gland complex glycogen content. We found several proteins which seem to be associated with the host immune response, most of which were up-regulated, however some were down-regulated, which may represent an important clue in understanding B. glabrata - S. mansoni compatibility., (Copyright © 2019 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Introduction: The increase in the use of electronic cigarettes (e-cigs) in young people and the lack of knowledge of the health effects of smoking in the short and long term are worrying. Although the oral cavity is the first to interact directly with the e-cig aerosol, studies on potential oral cavity lesions are still limited and there is some controversy about safety., Objective: To perform a systematic review to evaluate the adverse effects of e-cigs on oral health., Materials and Methods: The research was conducted using Cochrane Library, Embase, PubMed, and Web of Science. The research was limited to articles in English, Portuguese, and Spanish, published between January 2003 and November 2018. The research question was formulated according to the population, intervention, comparison, outcome (PICO) strategy. The quality of the methodology of each study was evaluated following the guidelines described in the Risk Of Bias In Non-randomized Studies - of Interventions (ROBINS-I) tool., Results: The initial search resulted in 432 articles, of which only eight were included for analysis. Periodontal and peri-implant clinical and radiographic parameters (plaque index, clinical attachment loss, probing depth, peri-implant bone loss, and radiographic bone level) are worse, and proinflammatory cytokine levels are higher among electronic and conventional cigarette smokers than among nonsmokers. Bleeding on probing was higher in nonsmokers than in conventional cigarette smokers and e-cig users. Nine different lesions of the oral mucosa were detected, with nicotinic stomatitis, hairy tongue, and angular cheilitis being more prevalent in e-cig consumers., Conclusion: The results suggest that e-cigs are less harmful than conventional cigarettes. However, there is also a greater susceptibility of e-cig consumers to developing alterations in oral biological tissues than ex-smokers or nonsmokers. There is still a clear need for the development of new studies., (Copyright © 2019 Elsevier Inc. All rights reserved.)

When exposure of the pulp to external environment occurs, reparative dentinogenesis can be induced by direct pulp capping to maintain pulp tissue vitality and function. These clinical situations require the use of materials that induce dentin repair and, subsequently, formation of a mineralized tissue., Objective: This work aims to assess the effect of tricalcium silicate cements and mineral trioxide aggregate cements, including repairing dentin formation and inflammatory reactions over time after pulp exposure in Wistar rats., Methodology: These two biomaterials were compared with positive control groups (open cavity with pulp tissue exposure) and negative control groups (no intervention). The evaluations were performed in three stages; three, seven and twenty-one days, and consisted of an imaging (nuclear medicine) and histological evaluation (H&E staining, immunohistochemistry and Alizarin Red S)., Results: The therapeutic effect of these biomaterials was confirmed. Nuclear medicine evaluation demonstrated that the uptake of 99mTc-Hydroxymethylene diphosphonate (HMDP) showed no significant differences between the different experimental groups and the control, revealing the non-occurrence of differences in the phosphocalcium metabolism. The histological study demonstrated that in mineral trioxide aggregate therapies, the presence of moderate inflammatory infiltration was found after three days, decreasing during follow-ups. The formation of mineralized tissue was only verified at 21 days of follow-up. The tricalcium silicate therapies demonstrated the presence of a slight inflammatory infiltration on the third day, increasing throughout the follow-up. The formation of mineralized tissue was observed in the seventh follow-up day, increasing over time., Conclusions: The mineral trioxide aggregate (WhiteProRoot®MTA) and tricalcium silicate (Biodentine™) present slight and reversible inflammatory signs in the pulp tissue, with the formation of mineralized tissue. However, the exacerbated induction of mineralized tissue formation with the tricalcium silicate biomaterial may lead to the formation of pulp calcifications.

(1) Background: Recently, tricalcium silicate cements, such as Biodentine™, have emerged. This biomaterial has a calcium hydroxide base and characteristics like mineral aggregate trioxide cements, but has tightening times that are substantially more suitable for their application and other clinical advantages. (2) Methods: A retrospective clinical study was conducted with 20 patients, which included a clinical evaluation of the presence or absence of pulp inflammation compatible symptoms, radiographic evaluation of the periapical tissues, and structural alterations of the coronary restoration that supports pulp capping therapies with Biodentine™ and WhiteProRoot ® MTA. (3) Results: This clinical study revealed similar success rates between mineral trioxide cement and tricalcium silicates cements at 6 months, with 100% and 95% success rates, respectively. There were no statistically significant differences between both biomaterials and between these and the various clinical circumstances, namely the absolute isolation of the operating field, exposure size, the aetiology of exposure, and even the type of restorative material used. (4) Conclusions: Biodentine™ demonstrated a therapeutic effect on the formation of a dentin bridge accompanied by slight inflammatory signs, with a high clinical success rate, indicating the possibility of its effective and safe use in dental pulp direct capping in humans, similar to the gold standard material.

Objectives: To compare the treatments used to treat dentin hypersensitivity (DH), based on its efficacy and effect duration., Methods: Medline/PubMed, Cochrane Library, EMBASE and ClinicalTrials were searched for articles published between 1 January 2008 and 14 November 2018, in English, Portuguese or Spanish, reporting clinical trials, completed and with results. This systematic review protocol was registered in PROSPERO, number CRD42019121986., Results: Seventy-four randomised clinical trials were included in the systematic review, reporting patients from 16 to 65 years old, with a clinical diagnosis of DH, that evaluate the efficacy of a desensitising product, compared to pre-treatment, used the evaporative method stimulation and the visual analogue scale. These studies evaluated 5366 patients and at least 9167 teeth. Seven follow-up periods were considered corresponding to an immediate, medium or long-time effect. Sixty-six studies were included in the quantitative synthesis. Glutaraldehyde with HEMA, glass ionomer cements and Laser present significant immediate (until 7 days) DH reduction. Medium-term (until 1 month) reduction was observed in stannous fluoride, glutaraldehyde with HEMA, hydroxyapatite, glass ionomer cements and Laser groups. Finally, long-term significant reduction was seen at potassium nitrate, arginine, glutaraldehyde with HEMA, hydroxyapatite, adhesive systems, glass ionomer cements and LASER., Conclusions: All active ingredients show efficacy in DH reduction in different follow-up times. Only in-office treatments are effective in immediate DH reduction, maintaining its efficacy over time. For long-time effects, at-home treatments can also be used. More standardised evaluation protocols should be implemented to increase the robustly of the results., (© 2019 John Wiley & Sons Ltd.)

This work describes a method to fabricate three-dimensional paper microfluidic devices in one step, without the need of stacking layers of paper, glue, or tape. We used a nontransparent negative photoresist that allows patterning selectively (vertically) the paper, creating systems of two or three layers, including channels. To demonstrate the capabilities of this methodology, we designed, fabricated, and tested a six-level diluter. The performance of the device was also simulated using a simple numerical model implemented in the program PETSc-FEM. The resulting μPAD is small (1.6 cm × 2.2 cm), inexpensive, requires low volumes of sample (5 μL), and is able to perform mixing and dilution in 2 min.

Background: Traditional medicine has an important role in local communities, who use plants in the treatment of various diseases. The research of traditional uses of medicinal plants allows us to document and analyze ethnopharmacological practices. This paper reports on an ethnobotanical survey that was conducted in the Protected Landscape of the "Serra de Montejunto", a Portuguese area in the west of the Iberian Peninsula, where these studies were nonexistent., Methods: The information was obtained through semi-structured ethnobotanical interviews with 78 informants, who were selected from several zones from the study area to have a representative of the entire landscape, during 2014. Local medicinal uses of plants were identified and grouped into 10 categories through data analysis, in quantitative indices such as the relative frequency citation (RFC), the cultural importance index (CI), and the informant consensus factor (F IC ). These were used to evaluate the importance of medicinal plants to the locals., Results: In the fieldwork, we found 105 taxa used as medicinal plants which belong to 46 families, where Rosaceae, Asteraceae, Fabaceae, and Lamiaceae are the ones with more diversity. The plants were grouped into 10 categories, where the digestive category is the most cited, with 54 taxa, and the ophthalmological category is the less cited, with only one taxon. Leaves and aerial parts are the components most used. Infusion is the most reported form of preparation, along with the oral administration. Most plants referred in this study are still in use today; only 17 are no longer used at the present time because habits have changed. A catalog of medicinal plants was also drawn up., Conclusion: This work enabled us to explore once more our experiences and memories as well as the ancestral use of plants with the goal of expanding ethnopharmacological knowledge. The absence of ethnobotanical studies in this region led us to gather information about useful plants and their applications and benefits. This research helps in the conservation effort of the collective knowledge of medicinal plants for future generations. However, a detailed analysis by body system is still required.

(1) Background: Bisphosphonate-related osteonecrosis of the jaw (BRONJ) is one of the most often seen side effects in patients treated with nitrogen-containing bisphosphonates (BPs), a post-surgical non-healing wound condition. Since calcium phosphate (CP) compounds are able to adsorb zoledronate (ZOL) when used as a drug delivery vehicle, we aimed to verify if these ceramics might have a potential protective effect for soft tissues surrounding surgical osseous wounds. (2) Methods: The chemical reaction between ZOL and CP compounds was evaluated through ultraviolet-visible spectroscopy and elemental analysis. A primary culture of human gingival fibroblasts (HGF) was established as a model to evaluate the cytotoxicity of the association of ZOL (5-500 μM) and of ZOL/biphasic calcium phosphates (BCP). Metabolic activity, cell viability, types of cell death, the cell cycle through, and the migration ability of human gingival fibroblasts were evaluated. (3) Results: ZOL was adsorbed by biphasic calcium phosphate compounds in an aqueous solution. The HGF were sensitive to ZOL toxicity; nevertheless, ZOL/BCP showed a significant protective effect regarding metabolic activity, cell viability, and cell migration. (4) Conclusions: BCP interaction with ZOL reduces or abolishes its toxicity in HGF. This finding represents a potential solution for BRONJ in the case of patients undergoing therapy with ZOL.

(1) Background: Bisphenol A (BPA) based dental resins are commonly used in preventive and reparative dentistry. Since some monomers may remain unpolymerized in the application of dental resin, they dissolve in the saliva. (2) Methods: The literature search was carried out in Pubmed, Cochrane and Embase databases. Randomized controlled trials, cohort studies and case-control studies that evaluated BPA levels in human urine, saliva and/or blood were included. (3) Results: The initial search had 5111 results. A total of 20 studies were included in the systematic review. Most studies showed an increase of the levels of bisphenol A 1 h after treatments with composite resins and dental sealants. One week after treatments the levels were decreased. (4) Conclusions: Some clinical precautions should be taken to decrease the release of BPA, namely the use of rubber dam, the immediate polishing of all resins used, or the use of glycerin gel to avoid non-polymerization of the last resin layer, and mouthwash after treatment. Another preventive measure in addition to the above-mentioned is the use of the smallest possible number of restorations or sealants, a maximum of four per appointment. These measures are even more important in children, adolescents and pregnant women.

(1) Background: When pulp exposure occurs, reparative dentinogenesis can be induced by direct pulp capping to maintain the vitality and function of the tissue. The aim of this work was to assess the cytotoxicity and bioactivity of three different direct pulp capping materials, calcium hydroxide (Life ® ), mineral trioxide aggregate (WhiteProRoot ® MTA) and calcium silicate (Biodentine ™ ), in an odontoblast-like mouse cell line (MDPC-23). (2) Methods: Metabolic activity was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test (MTT)assay, viability by the sulforhodamine B (SRB) assay, and the type of death and cell cycle analysis by flow cytometry. Alkaline phosphatase was evaluated by polymerase chain reaction (PCR), and dentin sialoprotein expression was assessed by immunocytochemistry. Mineralization was determined by the Alizarin Red S colorimetric assay and quantified by spectrophotometry. (3) Results: Life ® induced a decrease in metabolic activity and viability, which is associated with an increase cell death. WhiteProRoot ® MTA and Biodentine™ induced similar effects in cytotoxicity assays, with an increase in the expression of dentin sialoprotein (DSP) and formation of mineralized deposits, especially with Biodentine™. (4) Conclusions: The results of WhiteProRoot ® MTA confirm its indication for these therapies, justifying its recognition as the "gold standard". Biodentine™ may be an alternative, since they promote the same cellular response that mineral trioxide aggregate (MTA) does.

The incorporation of functional monomers in dental adhesive systems promotes chemical interaction with dental substrates, resulting in higher adhesion forces when compared to micromechanical adhesion only. The 10-MDP monomer, whose chemical structure allows for a polar behavior which is favorable to adhesion, also promotes the protection of collagen fibers through the formation of MDP-calcium salts. This systematic review aimed to characterize the interface created by 10-MDP containing adhesive systems through an evaluation of the following parameters: Formation of nano-layered structures, capacity to produce an acid-base resistant zone, and adhesion stability. The research was conducted using PubMed, Cochrane Library, Web of Science and Embase, limited to English, Spanish, and Portuguese articles. The research was done according to the PICO strategy. The 10-MDP monomer has the capacity to produce an acid-base resistant zone on the adhesive interface, which increases the response to acid-base challenges. The adhesion established by these systems is stable over time. To have the best of these adhesive solutions, a scrubbing technique must be used to apply the adhesive system on dental substrates, in order to improve monomers infiltration and to create a stable bond. Time must be given for the solution to infiltrate, hybridize and form the MDP-Ca, improving adhesive stability.

Depression is the largest cause of disability worldwide, affecting 350 million people. Notwithstanding that clinical trials demonstrate antidepressants efficacy, the efficient response can vary individually concerning therapeutic dosage. Although important, plasma levels monitoring remains an analytical challenge whereas clean-up and pre-concentration represent critical steps. Therefore, this study aims to develop, optimize and validate a method for fluoxetine determination in human plasma, employing a laboratory-made device consisting of a PDMS stir bar sorptive for extraction, coupled with high-performance liquid chromatography-fluorescence detection (SBSE-HPLC-FD). Optimization involved sorption-desorption steps. For sorption, temperature and time were assessed by factorial and central composite design approaches, taking into account the desirability and the response surface results, with stirring speed also examined. For desorption kinetics and ultrasonic and magnetic stirring mode were evaluated. The proposed method after validation was robust, linear (25.00-1000.00 ng mL -1 , R 2  > 0.98) and presented good intra- (RSD 4.18%) and inter-day-assay (RSD 11.60%) precision and accuracy (recovery 109.60%), allowing reliable quantitation without interference. The method was successfully applied to real samples. SBSE-HPLC-FD could represent a feasible alternative with good cost-benefit for low-volume samples and therapeutic drug monitoring, as well as contributing to correlation studies between plasma fluoxetine levels and clinical response, which is still little studied., (© 2018 John Wiley & Sons, Ltd.)

The present work aims at evaluating the potential gains derived from partially replacing calcium in resorbable β-tricalcium phosphate (β-TCP) by two different molar percentages of strontium (5, 10) and zinc (1, 2), concomitantly with a fixed molar percentage (0.5) of manganese. Synthetic granular composite bone filling grafts consisting of doped β-TCP and an alkali-free bioactive glass were prepared and implanted in ~4 mm diameter bone defects drilled in the calvaria of Wistar rats used as animal models. The animals were sacrificed after 9 weeks of implantation and the calvaria was excised. Non-manipulated bone was used as positive control, while empty defects were used as a negative control group. The von Kossa staining revealed an enhanced new bone formation with increasing doping levels, supporting the therapeutic effects exerted by the doping elements. The percentage of newly formed bone was similar when the defects were filled with autologous bone, BG (previous results) or 3TCP2/7BG, which indicates that the latter two are excellent candidates for replacement of autologous bone as bone regeneration material. This finding confirms that doping with suitable doses of therapeutic ions is a good strategy towards transposing the bone graft materials to biomedical applications in humans.