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Objectives This article evaluated the effect of substrates rigidities on the post-fatigue fracture resistance of adhesively cemented simplified restorations in lithium disilicate glass ceramic. Methods Precrystalized computer-aided design/computer-aided manufacturing ceramic blocks were processed into disc-shaped specimens (n = 10, Ø = 10 mm), mimicking a simplified restoration at two thicknesses (0.5 and 1.0 mm). Thereafter, the discs were cemented onto different base substrates (dentin analogue [control], dentin analogue with a central core build-up of resin composite [RC], or glass ionomer cement [GIC]). The specimens were subjected to mechanical cycling in a chewing simulator (100 N, 1 × 106 cycles, 4 Hz) and then subjected to thermocycling aging (10,000 cycles, 5/37/55°C, 30 seconds). After the fatigue protocol, the specimens were loaded until failure (N) in a universal testing machine. Finite element analysis calculated the first principal stress at the center of the adhesive interface. Results The results showed that “restoration thickness,” “type of substrate,” and their interaction were statistically significant (one-way analysis of variance; p Conclusion More flexible base material reduces the fracture load and increases the stress magnitude of adhesively cemented lithium disilicate restorations regardless the ceramic thickness. Therefore, more rigid substrates are suggested to be used to prevent restoration mechanical failures.

Zika virus (ZIKV) emerged as a global public health concern due to its relationship with severe neurological disorders. Non-structural (NS) proteins of ZIKV are essential for viral replication, regulatory function, and subversion of host responses. NS2B is a membrane protein responsible for the regulation of viral protease activity. This protein has transmembrane domains critical for the localization of viral protease to the endoplasmic reticulum membrane and a hydrophilic domain essential for folding, recruitment, and protease activity. Therefore, NS2B is considered a cofactor of viral protease which processes viral polyprotein and is essential for virus replication, making it an attractive antiviral drug target. Here, we report the backbone 1H, 15N, 13C resonance assignments of the full-length NS2B by high-resolution NMR. The backbone assignment will be necessary for determining the three-dimensional structure and backbone dynamics of NS2B, interaction mapping and screening potential of antiviral drugs against ZIKV and related pathogenic flaviviruses.

To study the influence of peritoneal dialysis (PD) on the salivary metabolite profile of children and adolescents with renal failure. Healthy children/adolescents (n = 31; mean age: 12.18 ± 3.76) and children/adolescents subjected to PD (n = 12; mean age: 10.10 ± 4.25) were recruited. Oral health status assessed by the dmft/DMFT and Volpe-Manhold calculus indices. The 1H spectra were acquired in a 600-MHz Bruker nuclear magnetic resonance spectrometer and were subjected to multivariate analysis using partial least squares discriminant analysis (PLS-DA), orthogonal PLS-DA (O-PLS-DA), and univariate analysis through chi-square and t tests (SPSS 20.0, IL, USA), with a significance level of p 0.05, t test). PLS-DA and O-PLS-DA were able to distinguish both groups (ACC = 0.85, R2 = 0.80, Q2 = 0.15). Salivary metabolites decrease in creatine, propionate, and sugar levels in the PD group and an increase in creatinine, butyrate, and lactate levels when compared with the healthy group. Children and adolescents subjected to PD have a different salivary metabolic profile from that of their healthy subjects. Complications of peritoneal dialysis procedures could be monitored by proper knowledge of saliva characteristics as predictors of peritonitis-related outcome. The use of metabolomics in pediatric nephrology may be an innovative methodology for the early diagnosis and monitoring of kidney diseases.

Evaluate the cytocompatibility of Calen®/ZO, Calcicur®, Vitapex®, Endoflas®, and zinc oxide/eugenol-based (ZOE) root canal pastes (RCP) to human primary osteoblasts (HPO) through a simplified model for primary teeth. The model employed pipette tips filled with 0.037 g of paste, exposed to 185 µL of culture medium for 24 h (n = 6). Release of components was analysed by Proton Nuclear Magnetic Resonance Spectroscopy (1H-NMR). HPO were exposed to conditioned media for 24 h. Cell viability was assessed by cell density and metabolic activity, and release of interleukin 6 (IL-6), vascular endothelial growth factor (VEGF) and fibroblast growth factor (bFGF) by immunological assay. Physicochemical properties and antimicrobial efficacy were also evaluated. 1H-NMR spectra analysis showed similarity between ZOE, Endoflas®, Calcicur®, and Vitapex® compared to Calen®/ZO and positive control, which showed distinct released components. Calen®/ZO and Calcicur® exhibited high alkaline pH in all periods and showed similar solubility. Calen®/ZO, ZOE, and Vitapex® showed similar flow rate. Calen®/ZO, Calcicur®, and Vitapex® did not exhibit antimicrobial efficacy. Calen®/ZO presented cytotoxicity (p

The purpose of this critical review is to assess if children and adolescents with hyposalivation are more affected by dental caries than those with normal flow rate.A literature search was performed using keywords and MeSH terms related to hyposalivation and dental caries in the Medline/PubMed, Web of Science, Cochrane Library, Scopus, LILACS/BBO databases and in gray literature without language or date restrictions until March 2022. Observational studies that accessed the presence of dental caries in patients up to 18 years-old with hyposalivation and compared with a control group (normal salivation rate) were considered eligible. The results from search were imported to EndNote Web, where duplicates were removed followed by title/abstract and full text analysis.A total of 12,236 non-duplicated studies were found and 14 fulfilled the criteria and were included in the present review, 9 cross-sectional and 5 cohorts. Stimulated salivary flow rates were assessed in 3644 participants, aged 3-17 years. Three cohort and three cross-sectional studies observed association between low salivary flow rates and the presence of dental caries, while the other 9 included articles did not verify this association. However, the absence of a standard criteria for the hyposalivation classification in young patients was observed and brough light to this important limitation among the studies.The salivary flow rate estimation for caries risk assessment must be the target of further studies to make possible and reliable, homogeneous, and unbiasedly assessment of the association between hyposalivation and dental caries in young patients.

Thioredoxins are ubiquous proteins with 2 cysteines at the active site. The isoform 1 of Thioredoxin from Saccharomyces cerevisae (Trx1) has six sulphur aminoacids, two cysteines and four methionines. In this work we performed the replacement of cysteines by selenocysteines by growth of a transformed celular expression vector E. coli BL21-DE3 in selenocysteine containing culture medium. The Maldi-TOF spectra of Seleno/Sulphur substituted Trx1 revealed six component peaks with 46-48 Da range between them, that is the isotopic Seleno-Sulfur difference, showing the replacement of the Cysteines and Methionines to Selenocysteines and Selenomethionines. The Maldi-TOF spectra of the peptides derived from Trypsin digestion of the purified Thioredoxin (peptide mass fingerprint) show Selenocysteine and Selenomethionine containing peptides. Therefore we are demonstrating that cystein can be replaced by selenocystein and be metabolically converted to selenomethionine during Trx1 heterologous translation. Furthermore, the Maldi-TOF spectra are showing the presence of the most abundant isotopes of selenium inserted in the peptides containing cysteine and methionine, derived from the Trx1 digestion. The one dimensional 77Se–1H heteronuclear multiple quantum coherence NMR spectroscopy (1D-HMQC) for reduced Seleno substituted Trx1 (Se-Trx1), revealed three ressonance lines for 1Hβ1 from Selenocysteines 30 and 33, between 1.6 and 2,0 ppm. The bidimensional HMQC spectra (2D-HMQC) of the reduced Se-Trx1 show the 77 Se ressonance signal in 178 ppm, coupled with 1Hβ1 and 1Hβ2 lines between 2.1 and 1.8 ppm. The 1D-HMQC for oxidized Trx1 revealed the only one broad resonance in 2.6 ppm probably relative to the 1Hβ1 prótons. The 2D-HMQC spectrum of oxidized protein shows a higher chemical shift of selenocysteine 77Se (832 ppm) if compared to reduced state (178 ppm). Together these data are showing that the protocol of Se – S substitution developed here is a efficient method to label the active site of Thioredoxin 1 with a broad band chemical shift atom 77Se. Furthermore the large spectral window of the 77Se NMR detected between reduced and oxidized states of the Thioredoxin 1 shows that this atom is an excellent probe for accessing oxidative states and probably the conformational dynamics of the active site of the Se-Trx1.

The aim of this study was to characterize the salivary metabolomic profile in adolescents with juvenile systemic lupus erythematosus (jSLE). A total of 24 adolescents with jSLE (15.92 ± 2.06 years) and 12 systemically healthy controls (15.25 ± 2.7 years) were included in the study. Participants underwent rheumatologic testing and periodontal examination, with the recording of plaque index (PI), probing depth (PD), clinical attachment level (CAL), and bleeding on probing index (BPI). Unstimulated whole saliva was collected from both groups and stored at -80 ºC. The salivary proton nuclear magnetic resonance (1H-NMR) spectra were acquired in a spectrometer operating at 500 MHz. Partial least squared discriminant analysis (PLS-DA) and orthogonal PLS-DA (O-PLS-DA) were used for statistical analysis. Mean CAL and PI were significantly increased in the group with jSLE (p < 0.01). Patients with jSLE presented a significantly different salivary metabolic profile (accuracy = 0.54; R2 = 0.86; Q2 = -0.293), significantly higher salivary levels of N-acetyl sugars, and significantly reduced levels of phenylalanine, glycine, taurine, hydroxybutyrate, and valerate compared with healthy controls (p < 0.05). It is suggested that the salivary metabolomic profile analyzed by 1H NMR in patients with jSLE presents a different fingerprint that the systemically healthy subjects. Integrating the variation of metabolites with the identification of the metabolic pathways involved seems to provide a better understanding of the influence of systemic disease on salivary metabolites.

Zika virus (ZIKV) emerged as a global public health concern due to its relationship with severe neurological disorders. Non-structural (NS) proteins of ZIKV are essential for viral replication, regulatory function, and subversion of host responses. NS2B is a membrane protein responsible for the regulation of viral protease activity. This protein has transmembrane domains critical for the localization of viral protease to the endoplasmic reticulum membrane and a hydrophilic domain essential for folding, recruitment, and protease activity. Therefore, NS2B is considered a cofactor of viral protease which processes viral polyprotein and is essential for virus replication, making it an attractive antiviral drug target. Here, we report the backbone

Made available in DSpace on 2021-06-25T10:28:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-04-01 Background: This study aimed to evaluate surface treatments, adhesives and composites for repairing silorane based restorations. Material and Methods: One hundred and twenty truncated cones (2 mm smaller diameter and 4 mm larger diameter) made of silorane composite were divided in 12 groups according with the surface treatment (diamond bur and oxide aluminum abrasion), the adhesive (Adper Scothbond Multipurpose (3M ESPE), Ecusit (Voco) and Filtek P90 Adhesive (3M ESPE)). Each group was subdivided in two according with the composite used for repair (methacrylate and silorane). The repair was made with a second truncated cone build over the first one and bond strength assssed by tensile strength. Data were submitted to ANOVA 3-way and Tukey’s test (p

This study tested the null hypothesis that antihistamine-containing syrup does not change salivary metabolites in vitro and in vivo. For the in vitro experiments, saliva from 10 volunteers was mixed with a syrup or pill suspension of loratadine (1 mg/ml Claritin®, Schering-Plough, Rio de Janeiro, Brazil). For the in vivo experiment, 10 volunteers performed a mouth rinse with 10 mL of antihistamine syrup (Claritin®; Schering-Plough, Rio de Janeiro, Brazil) for 20 seconds and then discarded the rinse water. After 20 seconds, 5 mL of unstimulated whole saliva was spit into a plastic tube kept on ice. The protein profile of in vitro and in vivo experiments was analyzed using 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The samples were also analyzed by nuclear magnetic resonance (NMR) spectroscopy, followed by Principal Component Analysis and Wilcoxon test (p < 0.05). There were differences in salivary metabolites after syrup interaction. The salivary concentrations of acetate, n-caproate, arginine, glutamate, and lysine among other metabolites were reduced with the syrup in both in vivo and in vitro experiments (p < 0.05), but no differences were observed when the pill suspension was used (p > 0.05). Similar changes in metabolite profiles were observed in both in vitro and in vivo experiments. Electrophoresis revealed no difference in the salivary protein pattern. The null hypothesis was rejected because the intake of syrup medicine changes the salivary composition and influences oral homeostasis and susceptibility to oral diseases.

Made available in DSpace on 2022-04-28T19:45:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Objective: This study compared the clinical performance of large indirect restorations (IRs) versus direct restorations (DRs) in posterior teeth. Methods: Thirty subjects received two class II restorations (n = 60), one fabricated from a precured composite block (Grandio Blocs, VOCO) for the indirect technique (IT) and the other with light-cured composite (GrandioSO, VOCO) for the direct technique (DT). For IT, the restoration was created using the computer-aided design and computer-aided manufacturer (CAD/CAM) system. For DT, the material was applied light-cured by using a layering technique. All restorations were evaluated by using the World Dental Federation criteria. Results: Twenty-three subjects attended the 2-year recall, and 46 restorations were evaluated. No significant differences were detected between the techniques for most parameters analyzed (p > 0.05). For “color match” at 7 days and 6 months, better results were observed for the DT. In relation to the overall scores, all restorations were esthetically acceptable after 2 years, while 93.3% of DT and 90% of IT showed acceptable function. For biological scores, 96.67% of DRs and 100% of IRs was acceptable after 24 months. Considering all properties, the success rates were 93.3% for DRs and 90% for IRs. Conclusions: After 2 years, both restorations presented similar and good clinical behavior for all the properties analyzed. Clinical significance: Light-cured direct posterior composite restorations may perform similarly to indirect composite restorations made with precured CAD/CAM composite blocks up to 2 years. Department of Restorative Dentistry Institute of Science and Technology São Paulo State University—UNESP Department of Restorative Dentistry Institute of Science and Technology São Paulo State University—UNESP

Dissertação de Mestrado em Ciências Jurídico-Forenses apresentada à Faculdade de Direito Na presente dissertação será abordada a questão da dupla descrição no registo predial. Neste sentido, importa notar que este estudo é centrado na publicitação de direitos reais sobre imóveis, mais concretamente sobre prédios (rústicos, urbanos e mistos), e o seu regime de registo. Assim sendo, com a apreciação das características fundamentais do sistema registal português será possível descortinar algumas das suas fragilidades que, direta ou indiretamente, causam duplicações de registo. Serão, desta forma, abordadas temáticas especialmente controversas, como a inexistência de um cadastro eficaz, a extrema facilidade com que se inscrevem prédios nos serviços das finanças e a consequente insuficiência das descrições prediais. Além do mais, será analisada a insegurança gerada pelo regime dos meios de suprimentos de títulos, bem como a relação entre o registo e a usucapião, sendo também aprofundada a questão do fracionamento do prédio e a usucapião parcial. De igual modo, integrará uma apreciação do regime da habilitação de herdeiros e a falta de controlo da titularidade dos prédios em caso de falecimento, assim como o infortúnio abandono do princípio da competência territorial das conservatórias.Em súmula, evidencia-se uma excessiva facilidade do acesso ao registo, uma enorme desvalorização do sistema registal e um indigno desinteresse no seu aperfeiçoamento que, mantendo-se até aos dias de hoje, potenciam situações totalmente inconcebíveis. Como é, aliás, o caso da problemática aqui a tratar: a duplicação do mesmo prédio no registo, através da qual se publicita que duas pessoas distintas são, simultaneamente, proprietárias do mesmo prédio.Não obstante ser abordado o conflito entre os diferentes titulares registais, tal como as duas doutrinas sobre os critérios de resolução do mesmo, este trabalho centra-se na necessidade de encontrar mecanismos que possibilitem eliminar existência de duplas descrições. Neste ponto, constatamos que esta é uma tarefa extremamente difícil, pois tal implica uma rigorosa revisão de vários regimes diferentes. Mais concretamente, tornar mais exigente, quer o regime da justificação quer o da habilitação de herdeiros, voltar a consagrar a regra da competência territorial das conservatórias e investir no projeto SINERGIC enquanto cadastro verdadeiramente eficaz, o que, só por si, já ajudaria a minimizar o problema das inscrições matriciais. No meu entendimento, o mais urgente é regulamentar a forma como se inscrevem os prédios nos serviços das finanças, sendo imprescindível criar um regime próprio para as inscrições matriciais, através de um conjunto de requisitos que tenham que ser cumpridos a nível nacional por todos os serviços de finanças. This dissertation addresses the matter of duplicate report in register of deeds. In this sense, it is important to note that this study is centered on the publication of real property rights, more specifically on tenements (rustic, urban and mixed), and their registration system. Therefore, analysing the fundamental characteristics of the Portuguese registration system, will possibly reveal some of its weaknesses that, directly or indirectly, cause duplication of registration. In this way, especially controversial topics will be addressed, such as the lack of an effective register, the extreme ease with which tenements are registered in the Finance service and the consequent insufficiency of land descriptions. In addition, the insecurity generated by the title deeds supply regime will be analyzed, as well as the relationship between registration and adverse possession, and the issue of fractioned tenement and partial adverse possession will also be further investigated. Likewise, it will include an assessment of the regime for the qualification of heirs and the lack of control over the ownership of tenements / land in the event of death, as well as the regretable abandoning of the principle of territorial jurisdiction of the land registration offices.In short, there is an excessive ease in access to registration, a huge devaluation of the registration system and an unworthy lack of interest in its improvement, which, even today, lead to totally inconceivable situations. Such is the case of the situation to be addressed here: the duplication of the same tenement in the Land Registry, through which it is advertised that two different people are simultaneously owners of the same building.Despite addressing the conflict between the different registrants, as well as the two doctrines on the criteria for its resolution, this work focuses on the need to find mechanisms that make it possible to eliminate the existence of duplicate report. At this point, it is an extremely difficult task, as this implies a rigorous review of several different regimes. Specifically, to make both the justification regime and the qualification of heirs more thorough, re-establish the rule of territorial jurisdiction of land registration offices and invest in the SINERGIC project as a truly effective record which, in itself, would already help to minimize the problem of main office registrations.In my understanding, the most urgent thing is to regulate the way tenements / land are registered in the Finance service, being essential to create a specific regime for matrix registrations, through a set of requirements that must be complied at a national level by all services of Finance.

COVID-19 is an emergent, worldwide public health concern. Joint efforts have been made by scientific communities of various fields to better understand the mechanisms of action of SARS-CoV-2. The need to understand the pathophysiological fingerprint and pathways of this disease make metabolomics-related approaches an indispensable tool for properly answering concerns relating to disease course. Determination of the metabolomic profile may help to explain the heterogeneous spectra of COVID-19 clinical phenotypes and be useful in monitoring disease progression as well as therapeutic treatments. In this sense, saliva has proven to be a strategic biofluid, owing not only to its appeal as a noninvasive sampling method but also due to the capacity of the virus to invade epithelial cells of the oral mucosa and salivary gland ducts via ACE2 receptors. Accordingly, important changes in metabolism have been described relating to COVID-19, indicating that metabolomics may open new avenues for understanding the pathophysiology of this disease, especially via longitudinal study designs. Thus, we discuss the importance of comprehending the SARS-CoV-2 salivary metabolomic fingerprint and also highlight the situation of Brazil on the frontlines of the war against COVID-19.

Sugar cane defensin 5 (Sd5) is a small antifungal protein, whose structure is held together by four conserved disulfide bridges. Sd5 and other proteins sharing a cysteine-stabilized α-β (CSαβ) fold lack a regular hydrophobic core. Instead, they are stabilized by tertiary contacts formed by surface-exposed hydrophilic and hydrophobic residues. Despite excessive cross-links, Sd5 exhibits complex millisecond conformational dynamics involving all secondary structure elements. We used Carr-Purcell-Meiboom-Gill (CPMG) NMR relaxation dispersion (RD) measurements performed at different temperatures and denaturant concentrations to probe brief excursions of Sd5 to a sparsely populated "excited" state. Temperature-dependent CPMG RD experiments reveal that the excited state is enthalpically unfavorable, suggesting a rearrangement of stabilizing contacts formed by surface-exposed side chains and/or secondary structure, while the experiments performed at different denaturant concentrations suggest a decrease in accessible surface area of Sd5 in the excited state. The measured backbone

Background The N-terminal domain of Tetracenomycin aromatase/cyclase (TcmN), an enzyme derived from Streptomyces glaucescens, is involved in polyketide cyclization, aromatization, and folding. Polyketides are a diverse class of secondary metabolites produced by certain groups of bacteria, fungi, and plants with various pharmaceutical applications. Examples include antibiotics, such as tetracycline, and anticancer drugs, such as doxorubicin. Because TcmN is a promising enzyme for in vitro production of polyketides, it is important to identify conditions that enhance its thermal resistance and optimize its function. Methods TcmN unfolding, stability, and dynamics were evaluated by fluorescence spectroscopy, circular dichroism, nuclear magnetic resonance 15N relaxation experiments, and microsecond molecular dynamics (MD) simulations. Results TcmN thermal resistance was enhanced at low protein and high salt concentrations, was pH-dependent, and denaturation was irreversible. Conformational dynamics on the μs-ms timescale were detected for residues in the substrate-binding cavity, and two predominant conformers representing opened and closed cavity states were observed in the MD simulations. Conclusion Based on the results, a mechanism was proposed in which the thermodynamics and kinetics of the TcmN conformational equilibrium modulate enzyme function by favoring ligand binding and avoiding aggregation. General significance Understanding the principles underlying TcmN stability and dynamics may help in designing mutants with optimal properties for biotechnological applications.

Ixolaris is an anticoagulant protein identified in the tick saliva of Ixodes scapularis. Ixolaris contains 2 Kunitz like domains and binds to Factor Xa or Factor X as a scaffold for inhibition of the Tissue Factor (TF)/Factor VIIa (FVIIa). In contrast to tissue factor pathway inhibitor (TFPI), however, Ixolaris does not bind to the active site cleft of FXa. Instead, complex formation is mediated by the FXa heparin-binding exosite. Due to its potent and long-lasting antithrombotic activity, Ixolaris is a promising agent for anticoagulant therapy. Although numerous functional studies of Ixolaris exist, three-dimensional structure of Ixolaris has not been obtained at atomic resolution. Using the pET32 vector, we successfully expressed a TRX-His6-Ixolaris fusion protein. By combining Ni-NTA chromatography, enterokinase protease cleavage, and reverse phase HPLC (RP-HPLC), we purified isotopically labeled Ixolaris for NMR studies. 1D 1H and 2D 15N-1H NMR analysis yielded high quality 2D 15N-1H HSQC spectra revealing that the recombinant protein is folded. These studies represent the first steps in obtaining high-resolution structural information by NMR for Ixolaris enabling the investigation of the molecular basis for Ixolaris-coagulation factors interactions.

Ixolaris is a two-Kunitz tick salivary gland protein identified in Ixodes scapularis that presents sequence homology to TFPI (tissue factor pathway inhibitor). It binds to the coagulation enzyme factor Xa (FXa) or to its zymogen form, FX, and further inhibits tissue factor/FVIIa complex (extrinsic Xnase compex). Differently from TFPI, Ixolaris does not bind to the active site cleft of FXa. Instead, complex formation is mediated by the FXa heparin-binding exosite, which may also results in decreased FXa activity into the prothrombinase complex. The Ixolaris-FXa/FX complex formation has been characterized by using a combination of biophysical and biochemical technics although no structural data is currently available. In this study, we reported the NMR chemical shift assignment of Ixolaris, as a first step to further establishing the structure, dynamics and function relationship for this protein.

Background: Our aim was to compare salivary levels of secretory immunoglobulin A (s-IgA) in children with early childhood caries (ECCG) and those who are caries-free (CFG) and verify these levels in a follow-up period after restorative treatment. Materials and methods: We selected 46 systemically healthy children in the complete primary dentition period, who were allocated into two groups: CFG (n = 23) and ECCG (dmf-s > 0; n = 23). Unstimulated whole saliva was obtained at baseline from both groups and during the follow-up period (7 days, 1, 2 and 3 months) in the ECCG group. The s-IgA was measured using an ELISA assay, and total protein was assessed using the Bradford method. We also evaluated the flow rate (mL/min), Streptococcus mutans and Lactobacillus spp. counting using selective media plaques. The data were submitted to statistical analysis using the software SPSS 20.0 (SPSS Inc, IL, USA) with a confidence interval set at 95%. Results: Salivary s-IgA levels were higher in baseline of ECCG than in CFG (p0.05). However, we observed two different changes in s-IgA levels among participants: one group presented s-IgA reduction, and the other group demonstrated its maintenance. It was shown that patients from the ECCG group who presented a reduction in s-IgA levels during follow-up also showed a decrease in Streptococcus mutans and Lactobacillus spp. count (p0.05). Conclusions: The present data support the idea that children with early childhood caries present higher levels of s-IgA in saliva than caries-free children. The restorative dental treatment does not have a significant influence on salivary levels of this immunoglobulin during the follow-up period.

Introduction: Puerperal woman and new-born children are vulnerable and frequently have neglected health conditions. Aim: The objective is to describe the oral health and saliva profiles of women and their babies during the breastfeeding period, including breast milk. Materials and Methods: Forty-seven mothers were interviewed and demographic data were recorded. The mother-baby pairs underwent intraoral examination. The mothers were submitted to examination of oral mucosa, oral hygiene status by O’Leary, periodontal condition and caries (DMF-T: decayed, missing and filled teeth) in order to establish buccal conditions. The babies had their oral mucosa and teeth examined. Salivary samples of babies and mothers as well the breastmilk of mothers were collected and analysed by 1 H-NMR through a 600 MHz spectrometer. The data were analysed in a statistical program SPSS.21 (IBM Statistics). Results: The mothers’ mean age was 27-years-old and 53.9% of mothers were overweight. The oral condition revealed poor oral health: DMF-T=8.20, 72.4% had gingivitis and 62% had dental plaque. The babies presented 4.18% cases of oral candidiasis and 2.08% cases of Bohn nodules, with no caries. The salivary 1 H-NMR spectra from babies with more than six months of age showed increased levels of lactate, ethanol, acetate, propionate, N-butyrate and N-acetyl sugars and reduced levels of other sugars. The 1 H-NMR analysis of salivary samples from the mothers showed metabolites such as propionate, ethanol, lactate, acetate, butyrate and N-acetyl and sugar region. The 1 H-NMR breast milk demonstrated high quantity of lactose in a region of spectra characteristic from sugars. It was concluded that the mothers had low levels caries activity; however, though they had past dental caries history. This may have an impact on the oral health of their children. Conclusion: Our study focused on the oral health and saliva profiles of women and their babies during breastfeeding period. This data could design a preventive programme that would improve the oral health and quality of life.

Submitted by Ana Paula Valente Pinho Mafetano (anapmafetano@gmail.com) on 2019-01-24T16:42:53Z No. of bitstreams: 1 Dissertaçao Ana Paula Mafetano Correta com ficha.pdf: 62624817 bytes, checksum: ce24e752ac7f6cc3f0d4bd71524a268c (MD5) Approved for entry into archive by Silvana Alvarez null (silvana@ict.unesp.br) on 2019-01-25T16:43:59Z (GMT) No. of bitstreams: 2 mefetano_apvp_me_sjc_par.pdf: 373599 bytes, checksum: 874e4f46847780867c7b9a5436942d73 (MD5) mefetano_apvp_me_sjc_int.pdf: 62624817 bytes, checksum: ce24e752ac7f6cc3f0d4bd71524a268c (MD5) Made available in DSpace on 2019-01-25T16:43:59Z (GMT). No. of bitstreams: 2 mefetano_apvp_me_sjc_par.pdf: 373599 bytes, checksum: 874e4f46847780867c7b9a5436942d73 (MD5) mefetano_apvp_me_sjc_int.pdf: 62624817 bytes, checksum: ce24e752ac7f6cc3f0d4bd71524a268c (MD5) Previous issue date: 2018-12-18 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) O objetivo deste estudo foi avaliar a influência do tipo de agente espessante e da viscosidade do gel de peróxido de hidrogênio (PH) a 35%, na eficácia clareadora e microdureza do esmalte dental. Além disso, a decomposição da molécula de peróxido de hidrogênio dentro do gel clareador também foi verificada. Foram utilizados 110 dentes incisivos bovinos dos quais foram obtidos 324 espécimes em forma de discos de esmalte- dentina, com 4mm de diâmetro, utilizando-se broca trefina. A leitura da cor dos referidos espécimes foi realizada em um espectrofotômetro de reflectância, enquanto a microdureza do esmalte foi analisada através de um microdurômetro com indentador Knoop. Para verificação da concentração de peróxido foi utilizado o método de titulação utilizando o permanganato de potássio. Géis clareadores experimentais foram preparados, contendo cinco diferentes tipos de agentes espessantes (Ar - Aristoflex – Clariant, Ca - Carbopol 980 – Lubrizol, CMC – Carboximetilcelulose, Ae – Aerosil 200 - Evonik e Sa - Salcare – BASF), os quais foram adicionados em quantidades suficientes para produzir géis com viscosidades baixa (B - 50.000 cPs), média (M - 250.000 cPs) e alta (A – 1.000.000 cPs). Os espécimes foram divididos em dezoito grupos: CP (Controle positivo) - de peróxido sem adição de agente espessante, CN (Controle Negativo) –água ultra pura, CC (Controle Comercial) – gel de PH a 35% Whiteness HP-FGM, Ar/B, Ar/M, Ar/A; Ca/B, Ca/M e Ca/A; CMC/B, CMC/M, CMC/A, Ae/B, Ae/M, Ae/A, Sa/B, Sa/M e Sa/A. Os agentes clareadores foram aplicados sobre a superfície do esmalte por um total de 60 minutos. A cada 20 minutos, o agente clareador era trocado e uma amostra deste era coletada e as concentrações de peróxido de hidrogênio mensuradas. A microdureza do esmalte foi avaliada imediatamente após, sendo então as amostras imersas em saliva artificial por 7 dias, seguido pela leitura final da cor. Os dados foram analisados estatisticamente, empregando um nível de significância de 5%. Quanto ao percentual de redução da microdureza a ANOVA 2 fatores mostrou diferenças significativas para o tipo de espessante e viscosidade. Os resultados do testede Tukey foram: espessante – Ae 11,73(3,98)a, As 14,71(3,79)b, CMC 17,00(4,01)c, Ar 22,13(4,43)d e Ca 28,06(2,59); viscosidade – alta 15,39(8,52)a, média 18,56(5,42)b e baixa 22,22(4,40)c. Quanto ao DEab e DE00 não mostrou diferenças significativas. Com relação à concentração do peróxido de hidrogênio, as maiores reduções occorreram para o espessante Salcare. Concluiu-se que o tipo de espessante e viscosidade dos géis não influenciaram o efeito clareador, porém influenciaram na microdureza do esmalte. O tipo de espessante e viscosidade também influenciaram a concentração final do peróxido de hidrogênio, e o espessante. Sa apresentou o maior percentual de redução de peróxido. Todos os grupos apresentaram diferenças significativas e relação ao grupo CN. O grupo CA/A não apresentou diferença significativa em relação ao grupo CP e apenas o grupo Sa/M não apresentou diferença significativa em relação ao grupo CC. The aim of this study was to evaluate of the type of the agent thickener and viscosity of 35% hydrogen peroxide gel, in the bleaching efficacy and microhardness of dental enamel. In addition, the decomposition of the hydrogen peroxide molecule within the bleaching gel was also verified. We used 110 bovine incisor teeth of which 324 specimens were obtained in the form of enamel-dentin discs, with 4 mm diameter, using a trefin drill. The color reading of the aforementioned specimens was performed in a reflectance spectrophotometer, while the microhardness of the enamel was analyzed through a microdurometer with Knoop indenter. To verify the peroxide concentration, the titration method was used using potassium permanganate 0,1 N. Experimental bleaching gels were prepared, containing five different types of thickened agents (Ar – Aristoflex – Clariant, Ca – Carbopol 980 – Lubrizol, CMC – Carboxymethylcellulose, Ae – Aerosil 200 – Evonik and Sa – Salcare – BASF), which were added in sufficient quantities to produce gels with low viscosities (L – 50.000 cPs), Medium (M – 250.000 cPs) and High (H – 1.000.000 cPs). The specimens were divided into eighteen groups/subgroups: PC (positive control) – was applied a peroxide solution without adding thickened agent, NC (Negative control) – was applied ultrapure water, CC (Comercial control) and fifteen more groups, containing hydrogen peroxide at 35% associated with a specific thickened agent, in different viscosities, these being: Ar/L, Ar/Mand Ar/H (Aristoflex in low, medium and high viscosity); Ca/L, Ca/M and Ca/H (Carbopol in low, medium and high viscosity); CMC/L, CMC/M and CMC/H (Carboxymethylcellulose in low, medium and high viscosity); Ae/L, Ae/M and Ae/H medium and high viscosity). The bleaching agents were applied on the enamel surface for 60 minutes. Every 20 minutes, the bleaching agent was changed and a sample was collected and the hydrogen peroxide concentrations measued. The enamel microhardness was immersed in artificial saliva for 7 days, followed by the final color reading. Data were analyzed statistically, employing a significance level of 5%. Regarding the percentage of microhardness reduction the ANOVA 2 factors showed significant differences for the type of thickener and viscosity. The Tukey test results were: thickener – Ae 11.73(3,98)a, As 14.71(3.79)b, CMC 17.00(4.01)c, Ar 22.13(4.43) and Ca 28.06(2.59); viscosity – high 15.39(8.52)a, average 18.56(5.42)b and low 22.22(4.40)c. Regarding DEab and DE00 showed no significant differences. Regarding the concentration of hydrogen peroxide, the greatest reductions occurred for the Salcare thickener. It was concluded that the type of thickener and viscosity of the gels did not influence the bleaching effect, but influence the microhardness of the enamel. The type of thickener and viscosity also influenced the final concentration of hydrogen peroxide, and the thickener. Sa presented the highest percentage of peroxide reduction. All groups presented significant difference in relation to the CP group and only the Sa/M group did not present a significant difference in relation to the CC group.

Ixolaris is a potent tick salivary anticoagulant that binds coagulation factor Xa (FXa) and zymogen FX, with formation of a quaternary tissue factor (TF)/FVIIa/ FX(a)/Ixolaris inhibitory complex. Ixolaris blocks TF-induced coagulation and PAR2 signaling and prevents thrombosis, tumor growth, and immune activation. We present a high-resolution structure and dynamics of Ixolaris and describe the structural basis for recognition of FX. Ixolaris consists of 2 Kunitz domains (K1 and K2) in which K2 is strikingly dynamic and encompasses several residues involved in FX binding. This indicates that the backbone plasticity of K2 is critical for Ixolaris biological activity. Notably, a nuclear magnetic resonance–derived model reveals a mechanism for an electrostatically guided, high-affinity interaction between Ixolaris and FX heparin-binding (pro)exosite, resulting in an allosteric switch in the catalytic site. This is the first report revealing the structure-function relationship of an anticoagulant targeting a zymogen serving as a scaffold for TF inhibition.

Fag s 1 is a member of the Pathogen Related protein family 10 (PR-10) and can elicit cross-reaction with IgE antibodies produced against the birch pollen allergen Bet v 1. The Nuclear Magnetic Resonance (NMR) structure of Fag s 1 is presented along with its dynamic properties. It shares 66% identity with Bet v 1 and exhibits the expected three α-helices and seven β-sheets arranged as a semi-beta barrel and exposing the residues mapped as the Bet v 1 IgE epitope. The structural dynamics of Fag s 1 were monitored on the fast and intermediate timescales, using relaxation rates. The complex dynamics of Fag s 1 are closely related to the internal cavity, and they modulate IgE and ligand binding.

The water cavity of yeast thioredoxin 1 (yTrx1) is an ancestral, conserved structural element that is poorly understood. We recently demonstrated that the water cavity is involved in the complex protein dynamics that are responsible for the catalytically relevant event of coupling hydration, proton exchange, and motion at the interacting loops. Its main feature is the presence of the conserved polar residue, Asp24, which is buried in a hydrophobic cavity. Here, we evaluated the role of the solvation of Asp24 as the main element that is responsible for the formation of the water cavity in thioredoxins. We showed that the substitution of Asp24 with a hydrophobic residue (D24A) was not sufficient to completely close the cavity. The dynamics of the D24A mutant of yTrx1 at multiple time scales revealed that the D24A mutant presents motions at different time scales near the active site, interaction loops, and water cavity, revealing the existence of a smaller dissected cavity. Molecular dynamics simulation, along with experimental molecular dynamics, allowed a detailed description of the water cavity in wild-type yTrx1 and D24A. The cavity connects the interacting loops, the central β-sheet, and α-helices 2 and 4. It is formed by three contiguous lobes, which we call lobes A-C. Lobe A is hydrophilic and the most superficial. It is formed primarily by the conserved Lys54. Lobe B is the central lobe formed by the catalytically important residues Cys33 and Asp24, which are strategically positioned. Lobe C is the most hydrophobic and is formed by the conserved cis-Pro73. The central lobe B is closed upon introduction of the D24A mutation, revealing that independent forces other than solvation of Asp24 maintain lobes A and C in the open configuration. These data allow us to better understand the properties of this enzyme.