Mating behavior and oviposition of Palpita forficifera (Lepidoptera: Crambidae).

Palpita forficifera Munroe, 1959 (Lepidoptera: Crambidae) is considered the main pest of the olive tree (Olea europaea L., Oleaceae) in Brazil and Uruguay. The aim of this work was to study the mating and oviposition behavior of P. forficifera in the field and laboratory. In the field, the sex emitting the mating pheromone was determined and in the laboratory, the rate of emergence of males and females; the age, time and duration of mating; number of copulations and oviposition time of P. forficifera were recorded. The field results showed that it was possible to capture up to five males per trap in just one night in traps with the presence of female P. forficifera. Copulation occurs between the seventh and twenty-third day of life and is most frequent during the third and sixth hours of scotophase. The average duration of the first copulation was 174 min, with 35% of couples recopulating, and there were cases of up to five copulations. Oviposition times were concentrated between 20:00 and 02:00. The results obtained provide insight into the reproductive behavior of P. forficifera and are useful for future studies aimed at identifying the sex pheromone to improve monitoring of the pest in olive orchards.

Gaze entropy metrics for mental workload estimation are heterogenous during hands-off level 2 automation.

As the level of vehicle automation increases, drivers are more likely to engage in non-driving related tasks which take their hands, eyes, and/or mind away from the driving task. Consequently, there has been increased interest in creating Driver Monitoring Systems (DMS) that are valid and reliable for detecting elements of driver state. Workload is one element of driver state that has remained elusive within the literature. Whilst there has been promising work in estimating mental workload using gaze-based metrics, the literature has placed too much emphasis on point estimate differences. Whilst these are useful for establishing whether effects exist, they ignore the inherent variability within individuals and between different drivers. The current work builds on this by using a Bayesian distributional modelling approach to quantify the within and between participants variability in Information Theoretical gaze metrics. Drivers (N = 38) undertook two experimental drives in hands-off Level 2 automation with their hands and feet away from operational controls. During both drives, their priority was to monitor the road before a critical takeover. During one drive participants had to complete a secondary cognitive task (2-back) during the hands-off Level 2 automation. Changes in Stationary Gaze Entropy and Gaze Transition Entropy were assessed for conditions with and without the 2-back to investigate whether consistent differences between workload conditions could be found across the sample. Stationary Gaze Entropy proved a reliable indicator of mental workload; 92 % of the population were predicted to show a decrease when completing 2-back during hands-off Level 2 automated driving. Conversely, Gaze Transition Entropy showed substantial heterogeneity; only 66 % of the population were predicted to have similar decreases. Furthermore, age was a strong predictor of the heterogeneity of the average causal effect that high mental workload had on eye movements. These results indicate that, whilst certain elements of Information Theoretic metrics can be used to estimate mental workload by DMS, future research needs to focus on the heterogeneity of these processes. Understanding this heterogeneity has important implications toward the design of future DMS and thus the safety of drivers using automated vehicle functions. It must be ensured that metrics used to detect mental workload are valid (accurately detecting a particular driver state) as well as reliable (consistently detecting this driver state across a population).

Detecting subtle transcriptomic perturbations induced by lncRNAs knock-down in single-cell CRISPRi screening using a new sparse supervised autoencoder neural network.

Single-cell CRISPR-based transcriptome screens are potent genetic tools for concomitantly assessing the expression profiles of cells targeted by a set of guides RNA (gRNA), and inferring target gene functions from the observed perturbations. However, due to various limitations, this approach lacks sensitivity in detecting weak perturbations and is essentially reliable when studying master regulators such as transcription factors. To overcome the challenge of detecting subtle gRNA induced transcriptomic perturbations and classifying the most responsive cells, we developed a new supervised autoencoder neural network method. Our Sparse supervised autoencoder (SSAE) neural network provides selection of both relevant features (genes) and actual perturbed cells. We applied this method on an in-house single-cell CRISPR-interference-based (CRISPRi) transcriptome screening (CROP-Seq) focusing on a subset of long non-coding RNAs (lncRNAs) regulated by hypoxia, a condition that promote tumor aggressiveness and drug resistance, in the context of lung adenocarcinoma (LUAD). The CROP-seq library of validated gRNA against a subset of lncRNAs and, as positive controls, HIF1A and HIF2A, the 2 main transcription factors of the hypoxic response, was transduced in A549 LUAD cells cultured in normoxia or exposed to hypoxic conditions during 3, 6 or 24 h. We first validated the SSAE approach on HIF1A and HIF2 by confirming the specific effect of their knock-down during the temporal switch of the hypoxic response. Next, the SSAE method was able to detect stable short hypoxia-dependent transcriptomic signatures induced by the knock-down of some lncRNAs candidates, outperforming previously published machine learning approaches. This proof of concept demonstrates the relevance of the SSAE approach for deciphering weak perturbations in single-cell transcriptomic data readout as part of CRISPR-based screening.

Prevalence of urologic sequelae and bladder and bowel dysfunctions in patients with congenital Zika syndrome: A multicenter evaluation of the Zika virus bladder and bowel sequelae assistance network.

INTRODUCTION: Neurogenic bladder was first confirmed as a urological sequela of Congenital Zika Syndrome (CZS) in 2018. Further clinical-epidemiological evidence also confirmed neurogenic bowel dysfunction and cryptorchidism. To strengthen the care for these children, the Congenital Zika Virus Bladder and Bowel Sequelae Network (RASZ in Brazilian) was created, including six integrated centers in Brazil. This article represents the initial outcome of the efforts by RASZ. OBJECTIVE: To evaluate the prevalence of bladder and bowel dysfunction, cryptorchidism and other urological sequelae related to CZS in cohorts attended in six Brazilian states. STUDY DESIGN: Observational, prospective, multicenter study including children with CZS assisted in one of six RASZ collaborative centers between June 2016 and February 2023. Data were collected from patient's first assessment using the same protocols for urological and bowel evaluation. Categorical variables were analyzed by frequency of occurrence and numerical variables by mean, median, and standard deviation. The study was approved by the Research Ethics Committees of each center, all parents/caregivers provided written informed consent. RESULTS: The study included 414 children aged 2 months to 7 years (mean 2.77 years, SD 1.73), 227 (54.8 %) were male and 140 (33,8 %) referred urological and bowel symptoms on arrival. Prevalence of both urological and bowel sequelae was 66.7 %, 51 % of children aged 4 years and older had urinary incontinence (UI). UTI was confirmed in 23.4 % (two presented toxemia) and among males, 18.1 % had cryptorchidism. Renal ultrasonography, performed in 186 children, was abnormal in 25 (13.4 %), 7 had hydronephrosis. Among the 287 children who performed urodynamics, 283 (98.6 %) were altered: 232 had a lower bladder capacity, 144 a maximum bladder pressure of ≥40 cm H2O, and 127 did not satisfactorily empty their bladder. DISCUSSION: A higher prevalence of NLUTD, neurogenic bowel and cryptorchidism was confirmed in children with CZS. Early diagnosis and appropriate treatment, including a multidisciplinary approach, may reduce the risk of UTIs, UI and kidney damage. A limitation of the study was the inability of children to complete the protocol, specifically urodynamic evaluation, and ultrasonography. In both exams, the percentage of abnormal cases was higher than that expected in the normal population. CONCLUSION: A 66,7 % prevalence of combined urological sequelae and bladder-bowel dysfunction related to CZS was confirmed in patients evaluated in six Brazilian cohorts. The most frequent changes were related to NLUTD, neurogenic bowel, and cryptorchidism. Prevalence may be underestimated due to access restrictions to diagnostic tests.

Effect of whitening toothpastes and activated charcoal powder on enamel wear and surface roughness.

This study aimed to evaluate surface roughness (Sa), roughness profile (Rv), and enamel wear after brushing with different whitening toothpastes and charcoal powders. Sixty (n = 10) bovine enamel blocks (6 × 6 × 3 mm) were randomly distributed into six groups according to toothpaste type: regular toothpaste (CONT), toothpaste containing 2% hydrogen peroxide (HP), toothpaste containing titanium dioxide (TiO2), toothpaste containing charcoal (COAL), toothpaste containing charcoal and TiO2 (COAL+TiO2), and activated charcoal powder (COAL_PWD). Each block was subjected to 30,000 reciprocal cycles at a 1:3 proportion slurry. After brushing, the blocks were analyzed using an optical profilometer to determine Sa, Rv, and enamel wear. In addition, representative 3D images of each group and wear profiles were obtained. Sa was analyzed using generalized linear models followed by Bonferroni correction, whereas Rv was analyzed using one-way analysis of variance. After brushing, COAL and COAL+TiO2 showed higher Sa values than COAL_PWD. However, no significant difference was observed in Sa between whitening toothpaste and COAL_PWD, and CONT (p > 0.05). In addition, no differences were observed among the groups in Rv (p > 0.05). Conversely, enamel wear was higher for TiO2, COAL, COAL+TiO2, and COAL_PWD than for CONT. CONT showed the least enamel wear, whereas HP showed intermediate values. Representative 3D images and line profiles showed lower step-height and lower mean surface losses for the CONT and HP groups than for the other groups. Whitening toothpastes and COAL_PWD did not increase Sa or Rv compared with CONT, while CONT demonstrated lower enamel wear.

Convergent QSAR Models for the Prediction of Cruzain Inhibitors.

Chagas disease is a parasitosis caused by Trypanosoma cruzi. Cruzain, the major cysteine protease from T. cruzi, is an excellent therapeutic target in the search for antichagasic drugs. It is important in the role of cell invasion, replication, differentiation, and metabolism of the parasite. In this work, we developed and assessed multiple quantitative structure-activity relationship (QSAR) models for a set of 61 cruzain inhibitors. These models include two-dimensional (2D) QSAR, three-dimensional (3D) QSAR, such as comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA), and Hologram QSAR (HQSAR). In total, we generated 10 major and 114 minor model variations. Molecular docking was used to successfully align the molecules. All CoMFA and CoMSIA models, which incorporate multiple fields, demonstrated robustness in our analysis. Steric fields exhibited satisfactory convergence in the contour maps, while the electrostatic field converged into a single small region. The HQSAR model taking into consideration only Atoms and Connectivity, with fragment sizes ranging from two to five atoms, was considered the best of the HQSAR variations, despite exhibiting a higher level of deviance. In total, 78 model variations meet the minimum requirements to be considered acceptable. We found that using as few as five descriptors it is possible to obtain robust results with 2D-QSAR. Models such as Random Forest, Tree Ensemble, Linear Regression, and HQSAR are recommended for working with large data sets, while the 3D-QSAR models are intended to study the geometry of the ligands, to optimize them into new and better performing antichagasics. Virtual Screening of a set of hydrazones, guided by the top-performing models, identified promising candidates for experimental validation. Among them, dv007 and dv015 exhibited consistently high predicted pIC50 values (7.26 and 7.24, respectively), making them compelling candidates for further drug development.

Inhibition of SARS-CoV-2 Infection in Vero Cells by Bovine Lactoferrin under Different Iron-Saturation States.

Despite the rapid mass vaccination against COVID-19, the emergence of new SARS-CoV-2 variants of concern, such as omicron, is still a great distress, and new therapeutic options are needed. Bovine lactoferrin (bLf), a multifunctional iron-binding glycoprotein available in unsaturated (apo-bLf) and saturated (holo-bLf) forms, has been shown to exert broad-spectrum antiviral activity against many viruses. In this study, we evaluated the efficacy of both forms of bLf at 1 mg/mL against infection of Vero cells by SARS-CoV-2. As assessed with antiviral assays, an equivalent significant reduction in virus infection by about 70% was observed when either form of bLf was present throughout the infection procedure with the SARS-CoV-2 ancestral or omicron strain. This inhibitory effect seemed to be concentrated during the early steps of virus infection, since a significant reduction in its efficiency by about 60% was observed when apo- or holo-bLf were incubated with the cells before or during virus addition, with no significant difference between the antiviral effects of the distinct iron-saturation states of the protein. However, an ultrastructural analysis of bLf treatment during the early steps of virus infection revealed that holo-bLf was somewhat more effective than apo-bLf in inhibiting virus entry. Together, these data suggest that bLf mainly acts in the early events of SARS-CoV-2 infection and is effective against the ancestral virus as well as its omicron variant. Considering that there are no effective treatments to COVID-19 with tolerable toxicity yet, bLf shows up as a promising candidate.

BugSigDB captures patterns of differential abundance across a broad range of host-associated microbial signatures.

The literature of human and other host-associated microbiome studies is expanding rapidly, but systematic comparisons among published results of host-associated microbiome signatures of differential abundance remain difficult. We present BugSigDB, a community-editable database of manually curated microbial signatures from published differential abundance studies accompanied by information on study geography, health outcomes, host body site and experimental, epidemiological and statistical methods using controlled vocabulary. The initial release of the database contains >2,500 manually curated signatures from >600 published studies on three host species, enabling high-throughput analysis of signature similarity, taxon enrichment, co-occurrence and coexclusion and consensus signatures. These data allow assessment of microbiome differential abundance within and across experimental conditions, environments or body sites. Database-wide analysis reveals experimental conditions with the highest level of consistency in signatures reported by independent studies and identifies commonalities among disease-associated signatures, including frequent introgression of oral pathobionts into the gut.

Spatial and temporal dispersion of Doryctobracon areolatus (Szépligeti) (Hymenoptera: Braconidae) in orchards infested with Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae).

Doryctobracon areolatus (Szépligeti) is an endoparasitoid and promising fruit fly control agent. The objective of the study was to determine the spatial (horizontal and vertical) and temporal dispersion of D. areolatus in the field. To evaluate the horizontal and temporal dispersion, two peach orchards were selected. In each orchard, 50 points were marked at different distances from the central point, from where 4,100 couples of D. areolatus were released. Four hours after release, parasitism units (PU) (3 per point) were fixed to the trees at a height of 1.5 m from the ground. The PUs were composed of ripe apples artificially infested with second instar larvae of Anastrepha fraterculus (30 larvae/fruit). For the evaluation of vertical dispersion, in an olive orchard six points were selected (trees of ≈4 m in height). Each tree was divided into three heights in relation to the ground (1.17, 2.34, and 3.51 m). Doryctobracon areolatus was able to disperse horizontally at a distance >60 m from the release point. However, the highest parasitism rates [15-45% (area 1); 15-27% (area 2)] were observed up to 25 m. Higher percentages of parasitism and of recovered offspring occur in the first days after the release of the parasitoid (2 DAR). As for vertical dispersion, D. areolatus parasitized A. fraterculus larvae up to the highest attachment height of the evaluated PUs (3.51). The results showed the potential use of D. areolatus in the management of fruit flies in the field.

Lethal and sublethal effects of toxic bait formulations on Doryctobracon areolatus (Hymenoptera: Braconidae) and implications for integrated fruit fly management.

The use of toxic baits has become one of the main methods of management of fruit flies in Brazil. The application of toxic baits may cause side effects on the native parasitoid Doryctobracon areolatus (Hymenoptera: Braconidae). Based on the results, formulations made from the food attractants 3% Biofruit, 1.5% Ceratrap, 1.25% Flyral, 3% Isca Samaritá, 3% Isca Samaritá Tradicional, and 7% sugarcane molasses associated with the Malathion 1000 EC and the ready-to-use toxic bait Gelsura (containing the active ingredient alpha-cypermethrin) were classified as harmful (class 4) to D. areolatus (mortality > 85% at 96 HAE). In contrast, for toxic baits formulated with insecticide phosmet, the mortality ranged from 38% to 72%, classified as slightly harmful or moderately harmful. However, when phosmet was added to the 3% Samaritá Tradicional bait, the mortality was only 3.9% (class 1-harmless), similar to the toxicity observed for the Success 0.02 CB ready-to-use bait (0.24 g a.i. spinosad/l) (<5% mortality). Although toxic baits were formulated with spinosyn-based insecticides, all toxic bait formulations were classified as harmless or slightly harmful (<50% mortality) to D. areolatus, with the exception of 1.5% Ceratrap + spinetoram and 7% Sugarcane molasses + spinosad (≈ 60% mortality-moderately harmful). In addition, these formulations did not show sublethal effects in reducing the parasitism and emergence rate of the F1 generation of D. areolatus in A. fraterculus larvae. The results serve as a basis for the correct use of toxic food baits without affecting the biological control.

Efecto de diferentes condiciones y modos de aplicación sobre la fuerza de unión de los adhesivos a la dentinaEffect of different conditions and modes of application on bond strength of adhesives to dentine / Effect of different conditions and modes of application on bond strength of adhesives to dentine

Objective: To evaluate the microtensile bond strength in different dentine conditions (etched-E, non-etched-N, dry-D and wet-W) of a multimode adhesive (Scotchbond Universal-SU, 3M/ESPE) and a total etching adhesive (Ambar-AB, FGM) using a sonic device (Smart Sonic Device-SD, FGM). Material and methods: In this in vitro study, ninety six sound extracted human molars were divided into 12 groups (n=8) according to different dentine conditions and adhesive systems. Enamel was removed and the middle dentine surfaces were polished. Each adhesive system was applied according to the different dentine conditions, and composite resin blocks were incrementally built up and stored for 24 hours. Specimens were sectioned into sticks and bond strength data were analyzed with the Kruskal-Wallis test and Mann-Whitney U tests. Results: No effects of sonic application and were observed. In general, AB showed lower results compared to the SU. E and N conditions did not statistically affect the bond strength of SU groups. Dry dentine presented statistically superior bond strength values when compared to wet dentine for SU/E/SD group. Conclusion: Adhesion of dry dentine with multimode adhesive system may be superior to wet dentine with sonic application. The modes of application had no influence in bond strength of studied adhesives.

Objetivo: Evaluar la resistencia de la unión microtensil en diferentes condiciones de dentina (grabado-E, sin grabado-N, seco-D y húmedo-W) de un adhesivo multimodo (Scotchbond Universal-SU, 3M/ESPE) y un adhesivo de grabado total (Ambar-AB, FGM) utilizando un dispositivo sónico (Smart Sonic Device-SD, FGM). Material y Métodos: En este estudio in vitro, noventa y seis molares humanos extraídos sanos se dividieron en 12 grupos (n=8) de acuerdo con diferentes condiciones de dentina y sistemas adhesivos. Se eliminó el esmalte y se pulieron las superficies centrales de la dentina. Cada sistema adhesivo se aplicó de acuerdo con las diferentes condiciones de dentina, y los bloques de resina compuesta se acumularon de forma incremental y se almacenaron durante 24h. Las muestras se seccionaron en barras y los datos de resistencia de la unión se analizaron con la prueba de Kruskal-Wallis y la prueba de U de Mann-Whitney. Resultado: No se observaron efectos de la aplicación sónica. En general, AB mostró resultados más bajos en comparación con el SU. Las condiciones E y N no afectaron estadísticamente la fuerza de unión de los grupos SU. La dentina seca presentó valores de fuerza de adhesión estadísticamente superiores en comparación con la dentina húmeda para el grupo SU/E/SD. Conclusión: La adhesión de la dentina seca con un sistema adhesivo multimodo puede ser superior a la dentina húmeda con aplicación sónica. Los modos de aplicación no tuvieron influencia en la resistencia de la unión de los adhesivos estudiados.

Effect of whitening toothpastes and activated charcoal powder on enamel wear and surface roughness

Abstract This study aimed to evaluate surface roughness (Sa), roughness profile (Rv), and enamel wear after brushing with different whitening toothpastes and charcoal powders. Sixty (n = 10) bovine enamel blocks (6 × 6 × 3 mm) were randomly distributed into six groups according to toothpaste type: regular toothpaste (CONT), toothpaste containing 2% hydrogen peroxide (HP), toothpaste containing titanium dioxide (TiO2), toothpaste containing charcoal (COAL), toothpaste containing charcoal and TiO2 (COAL+TiO2), and activated charcoal powder (COAL_PWD). Each block was subjected to 30,000 reciprocal cycles at a 1:3 proportion slurry. After brushing, the blocks were analyzed using an optical profilometer to determine Sa, Rv, and enamel wear. In addition, representative 3D images of each group and wear profiles were obtained. Sa was analyzed using generalized linear models followed by Bonferroni correction, whereas Rv was analyzed using one-way analysis of variance. After brushing, COAL and COAL+TiO2 showed higher Sa values than COAL_PWD. However, no significant difference was observed in Sa between whitening toothpaste and COAL_PWD, and CONT (p > 0.05). In addition, no differences were observed among the groups in Rv (p > 0.05). Conversely, enamel wear was higher for TiO2, COAL, COAL+TiO2, and COAL_PWD than for CONT. CONT showed the least enamel wear, whereas HP showed intermediate values. Representative 3D images and line profiles showed lower step-height and lower mean surface losses for the CONT and HP groups than for the other groups. Whitening toothpastes and COAL_PWD did not increase Sa or Rv compared with CONT, while CONT demonstrated lower enamel wear.

Bovine Lactoferrin Induces Cell Death in Human Prostate Cancer Cells.

Bovine lactoferrin (bLf) is a multifunctional protein widely associated with anticancer activity. Prostate cancer is the second most frequent type of cancer worldwide. This study was aimed at evaluating the influence of bLf on cell viability, cell cycle progression, reactive oxygen species (ROS) production, and rate of apoptosis in the human prostate cancer cell line (DU-145). MTT assay and trypan blue exclusion were used to analyze cell viability. Morphological changes were analyzed through optical microscopy after 24 h and 48 h of bLf treatment. FITC-bLf internalization and cellular damage were observed within 24 h by confocal fluorescence microscopy. Cell cycle analyses were performed by flow cytometry and propidium iodide. For caspases 3/7 activation and reactive oxygen species production evaluation, cells were live-imaged using the high-throughput system Operetta. The cell viability assays demonstrated that bLf induces cell death and morphological changes after 24 h and 48 h of treatment compared to control on DU-145 cells. The bLf internalization was detected in DU-145 cells, G1-phase arrest of the cell cycle, caspase 3/7 activation, and increased oxidative stress on bLf-treated cells. Our data support that bLf has an important anticancer activity, thus offering new perspectives in preventing and treating prostate cancer.

The effect of information from dash-based human-machine interfaces on drivers' gaze patterns and lane-change manoeuvres after conditionally automated driving.

The goal of this paper was to measure the effect of Human-Machine Interface (HMI) information and guidance on drivers' gaze and takeover behaviour during transitions of control from automation. The motivation for this study came from a gap in the literature, where previous research reports improved performance of drivers' takeover based on HMI information, without considering its effect on drivers' visual attention distribution, and how drivers also use the information available in the environment to guide their response. This driving simulator study investigated drivers' lane-changing behaviour after resumption of control from automation. Different levels of information were provided on a dash-based HMI, prior to each lane change, to investigate how drivers distribute their attention between the surrounding environment and the HMI. The difficulty of the lane change was also manipulated by controlling the position of approaching vehicles in drivers' offside lane. Results indicated that drivers' decision-making time was sensitive to the presence of nearby vehicles in the offside lane, but not directly influenced by the information on the HMI. In terms of gaze behaviour, the closer the position of vehicles in the offside lane, the longer drivers looked in that direction. Drivers looked more at the HMI, and less towards the road centre, when the HMI presented information about automation status, and included an advisory message indicating it was safe to change lane. Machine learning techniques showed a strong relationship between drivers' gaze to the information presented on the HMI, and decision-making time (DMT). These results contribute to our understanding of HMI design for automated vehicles, by demonstrating the attentional costs of an overly-informative HMI, and that drivers still rely on environmental information to perform a lane-change, even when the same information can be acquired by the HMI of the vehicle.

Cadmium and lead transfer factors to kale plants (Brassica oleracea var. acephala) grown in mountain agroecosystem and its risk to human health.

The aim of the present study is to assess soil Cd and Pb contamination in kale (Brassica oleracea var. acephala) production sites in mountain agroecosystem, as well as its potential risk to human health. The study was carried out within 24 kale-production sites located in mountainous region of Rio de Janeiro (Brazil). Soil, plant, and fertilizer samples were collected in all assessed properties. Pseudo-total and bioavailable soil Cd and Pb content and their concentration and amount accumulated in plants were analyzed. The recorded results were used to calculate the pollution index. Risks to human health associated with kale consumption were assessed by comparing the limits set by the Brazilian regulating agency and by calculating estimated daily intake (EDI), noncarcinogenic target hazard quotient (THQ), and carcinogenic risk factor (CRF). Soil Cd and Pb enrichment was observed. Inappropriate management adopted in the assessed sites and terrain slope and the intensive use of mineral (phosphate and potassium) and organic (poultry litter) fertilizers were the main factors contributing to increase soil Cd and Pb pseudo-total and bioavailability contents. Most Cd and Pb contents in leaves, in natura, exceed the maximum values set by the Brazilian regulating agency. The present EDI, THQ, and CRF results recorded for these metals were within the tolerable ranges.

Two-year randomized clinical trial of different restorative techniques in non-carious cervical lesions and MMP activity in gingival crevicular fluid.

OBJECTIVES: To evaluate different restorative techniques for non-carious cervical lesions (NCCLs) and the activity of matrix metalloproteinases (MMPs) in gingival crevicular fluid. MATERIALS AND METHODS: Two hundred restorations were performed in 50 patients using resin composite restorative system without (I) and with selective enamel conditioning (II) and resin-modified glass-ionomer cement without (III) and with EDTA pretreatment (IV). Gingival crevicular fluid samples were collected in 15 patients. Restorations were evaluated using USPHS criteria at baseline and after 2 years. Percentages of MMP activity were assessed by zymography as a surrogate outcome. Equality tests of two proportions, logistic regression analysis, survival analysis, ANOVA repeated measures, and Fisher tests were used. RESULTS: No differences in clinical performance were found among groups. Group I had lower retention at 2 years than at baseline. Decreased alpha scores for marginal integrity and marginal discoloration were observed for all groups after 2 years. MMP-2 decreased after 1 year, and its activity increased back to the initial level after 2 years, mainly for groups I, II, and III. MMP-9 increased after 1 year, and it was reduced to the initial level after 2 years, mainly for group I. CONCLUSIONS: All restorative techniques performed similarly in NCCLs after 2 years with initial marginal defect alterations. MMP-2 reestablished its initial levels after 2 years, and MMP-9 had few alterations over time in crevicular fluid. Clinical relevance The different restorative techniques are equally successful in NCCLs after 2 years of clinical functioning and have similar effects on MMPs present in crevicular fluid.

Sodium trimetaphosphate combined to calcium as a strategy to improve dentin-bonding interface / Trimetafosfato de sódio associado ao cálcio como uma estratégia para melhorar a interface adesiva da dentina

Objective: The aim of this study was to evaluate the effect of STMP as biomimetic analog of dentin matrix on the dentin bond strength submitted to artificial cariogenic challenge over time. Material and Methods: The total number of teeth used in the experiment was 60 teeth, which were divided into 6 groups (n = 10). Of these total amount, 10 teeth were not submitted to the artificial cariogenic challenge (ACC), serving as control group (Sound Dentin - SD) while the other 50 were submitted to an ACC (7d/37ºC), being treated with treatment solutions according to each group: SD- deionized water/sound dentin, CD- deionized water/ artificial caries dentin, GIII- STMP, GIV- STMP + Ca(OH)2, GV- STMP + NaF, and GVI- NaF. After treatments (24h), the specimens were restored (Adper Single Bond Universal + Filtek Z250), to obtain resin­dentin sticks with a cross sectional area of 0.8mm2, approximately. Two-third of these sticks were stored in artificial saliva (37°C) for analyzes after 6 and 12 months. The 1/3 remains were subjected to µTBS test (baseline). Data were analyzed by two-way ANOVA and Tukey tests (p<0.05). Results: In general, the highest µTBS values were obtained in sound condition (SD), while the artificial caries condition (CD) determined minimum values. Groups treated with NaF (with or without STMP- GV and GVI) were not able to improve adhesion over time. Only the use of STMP + Ca(OH)2(GIV) improved the µTBS compared to the others caries-challenged dentin after 1 year. The adhesive failure pattern was predominant in all time. Conclusion: The use of the STMP associated with Ca(OH)2 seems to be a viable therapeutic strategy conciliating the biomimetizing capacity to the adhesive process satisfactorily even its performance is not superior to initial condition (AU)

Objetivo: O objetivo deste estudo foi avaliar o efeito do STMP como análogo biomimético da matriz dentinária na resistência de união à dentina submetida a desafio cariogênico artificial ao longo do tempo. Material e Métodos:foram utilizados um total de 60 dentes neste experimento, os quais foram divididos em 6 grupos (n = 10). Desse total, 10 dentes não foram submetidos ao desafio cariogênico artificial (DCA), servindo como grupo controle (Dentina Hígida - DH) enquanto os outros 50 foram submetidos ao DCA (7d / 37ºC), sendo tratados com soluções de tratamento específicas para cada grupo: DH- água deionizada / dentina hígida, DC- água deionizada / dentina submetida ao DCA, GIII- STMP, GIV- STMP + Ca(OH)2, GV- STMP + NaF e GVI- NaF. Após os tratamentos (24h), os corpos-de-prova foram restaurados (Adper Single Bond Universal + Filtek Z250), para obtenção de palitos de resina-dentina com área transversal de aproximadamente 0,8mm2. Dois terços desses palitos foram armazenados em saliva artificial (37°C) para análises após 6 e 12 meses. Os outros 1/3 foram submetidos ao teste µTBS (baseline). Os dados foram analisados por ANOVA a dois fatores e testes de Tukey (p <0,05). Resultados:Em geral, os maiores valores de µTBS foram obtidos em condição hígidas (DH), enquanto a condição subtmetidas ao DCA determinou os menores valores. Os grupos tratados com NaF (com ou sem STMP associado -GV e GVI) não foram capazes de melhorar a resistência de união, ao longo do tempo. Somente o uso de STMP + Ca (OH)2(GIV) melhorou o µTBS em comparação com as outras condições desafiadas por cárie após 1 ano. O padrão de falha adesiva foi predominante em todos os tempos. Conclusão: O uso do STMP associado ao Ca (OH)2 parece ser uma estratégia terapêutica viável conciliando a capacidade biomimetizante ao processo adesivo de forma satisfatória mesmo que seu desempenho não seja superior à condição inicial.(AU)

Influence of iron binding in the structural stability and cellular internalization of bovine lactoferrin.

Lactoferrin (Lf) is an iron-binding glycoprotein and a component of many external secretions with a wide diversity of functions. Structural studies are important to understand the mechanisms employed by Lf to exert so varied functions. Here, we used guanidine hydrochloride and high hydrostatic pressure to cause perturbations in the structure of bovine Lf (bLf) in apo and holo (unsaturated and iron-saturated, respectively) forms, and analyzed conformational changes by intrinsic and extrinsic fluorescence spectroscopy. Our results showed that the iron binding promotes changes on tertiary structure of bLf and increases its structural stability. In addition, we evaluated the effects of bLf structural change on the kinetics of bLf internalization in Vero cells by confocal fluorescence microscopy, and observed that the holo form was faster than the apo form. This finding may indicate that structural changes promoted by iron binding may play a key role in the intracellular traffic of bLf. Altogether, our data improve the comprehension of bLf stability and uptake, adding knowledge to its potential use as a biopharmaceutical.