Chronic oral disease burden at the first 1000 days: Intergenerational risk factors, BRISA cohort.

OBJECTIVE: To analyze multiple-causal models, including socioeconomic, obesity, sugar consumption, alcohol smoking, caries, and periodontitis variables in pregnant women with early sugar exposure, obesity, and the Chronic Oral Disease Burden in their offspring around the first 1000 days of life. METHODS: The BRISA cohort study, Brazil, had two assessments: at the 22nd-25th gestational weeks and during the child's second year (n = 1141). We proposed a theoretical model exploring the association between socioeconomic and pregnancy factors (age, smoking, alcohol, sugars, obesity, periodontitis, and caries) and child's variables (sugars and overweight) with the outcome, Chronic Oral Disease Burden (latent variable deduced from visible plaque, gingivitis, and tooth decay), using structural equation modeling. RESULTS: Caries and periodontitis were correlated in pregnant women. Addictive behaviors in the gestational period were correlated. Obesity (Standardized coefficient - SC = 0.081; p = 0.047) and added sugar consumption (SC = 0.142; p = 0.041) were observed intergenerationally in the pregnant woman-child dyads. Sugar consumption by the children (SC = 0.210; p = 0.041) increased the Chronic Oral Disease Burden. CONCLUSIONS: Poor caries and periodontal indicators were correlated in pregnant women and their offspring. Obesity and sugar consumption act intergenerationally. Oral health in early life may change life trajectory since the worst oral conditions predict main NCDs.

Prediction of deciduous teeth eruption in Brazilian children: A cross-sectional study nested in a prospective birth cohort (BRISA).

BACKGROUND: Dental eruption is part of a set of children´s somatic growth phenomena. The worldwide accepted human dental eruption chronology is still based on a small sample of European children. However, evidence points to some population variations with the eruption at least two months later in low-income countries, and local standards may be useful. So, this study aimed to predict deciduous teeth eruption from 12 months of age in a Brazilian infant population. METHODS: We developed a cross-sectional study nested in four prospective cohorts - the Brazilian Ribeirão Preto and São Luís Cohort Study (BRISA) - in a sample of 3,733 children aged 12 to 36 months old, corrected by gestational age. We made a reference curve with the number of teeth erupted by age using the Generalized Additive Models for location, scale, and shape (GAMLSS) technique. The explanatory variable was the corrected children´s age. The dependent variable was the number of erupted teeth, by gender, evaluated according to some different outcome distributional forms. The generalized Akaike information criterion (GAIC) and the model residuals were used as the model selection criterion. RESULTS: The Box-Cox Power Exponential method was the GAMLSS model with better-fit indexes. Our estimation curve was able to predict the number of erupted deciduous teeth by age, similar to the real values, in addition to describing the evolution of children's development, with comparative patterns. There was no difference in the mean number of erupted teeth between the sexes. According to the reference curve, at 12 months old, 25% of children had four erupted teeth or less, while 75% had seven or fewer and 95% had 11 or fewer. At 24 months old, 5% had less than 12, and 75% had 18 or more. At 36 months old, around 50% of the population had deciduous dentition completed (20 teeth). CONCLUSION: The adjusted age was an important predictor of the number of erupted deciduous teeth. This outcome can be a variable incorporated into children's growth and development curves, such as weight and height curves for age to help dentists and physicians in the monitoring the children's health.

Tandem Triplication 11p15.5-ICR1 (H19/IGF2) Detected by Microarray and Optical Genome Mapping in a Prenatal Beckwith-Wiedemann Case.

Optical genome mapping (OGM) appears as a new tool for matching standard cytogenetic methods (karyotype and microarray) into a single assay. The chromosomal region 11p15.5 harbours two differentially methylated regions, the imprinting centre regions 1 and 2 (ICR1, ICR2). Disturbances in both regions alter human growth and are associated with two imprinting disorders, Beckwith-Wiedemann (BWS) and Silver-Russell syndromes. Herein, we present a prenatal case with a triplication in 11p15.5, including the H19/IGF2 imprinted region, detected by microarray and OGM. A 30-year-old pregnant woman of 17 weeks of gestation was referred for prenatal karyotype and microarray study because of increased nuchal translucency, short femur, megabladder, hyperechogenic bowel, and renal ectasia. Microarray, OGM, and MS-MLPA were performed, and a tandem cis-triplication in 11p15.5 and hypermethylation of the ICR1 region, compatible with BWS was detected. OGM, with its power to detect all classes of structural variants, including copy number variants, at a higher resolution than traditional cytogenetic methods can play a significant role in prenatal care and management as a next-generation cytogenomic tool. This study further supports the hypotheses that the amplification/duplication-triplication of the H19/IGF2 region could be related to BWS if it is of paternal origin.

Metrics of care and cardiovascular outcomes in patients with ST-elevation myocardial infarction treated with pharmacoinvasive strategy: a decade-long network in a populous city in Brazil.

BACKGROUND: Pharmacoinvasive strategy is an effective myocardial reperfusion therapy when primary percutaneous coronary intervention (p-PCI) cannot be performed in a timely manner. METHODS: Authors sought to evaluate metrics of care and cardiovascular outcomes in a decade-long registry of a pharmacoinvasive strategy network for the treatment of ST-elevation myocardial infarction (STEMI). Data from a local network including patients undergoing fibrinolysis in county hospitals and systematically transferred to the tertiary center were accessed from March 2010 to September 2020. Numerical variables were described as median and interquartile range. Area under the curve (AUC-ROC) was used to analyze the predictive value of TIMI and GRACE scores for in-hospital mortality. RESULTS: A total of 2,710 consecutive STEMI patients aged 59 [51-66] years, 815 women (30.1%) and 837 individuals with diabetes (30.9%) were analyzed. The time from symptom onset to first-medical-contact was 120 [60-210] minutes and the door-to-needle time was 70 [43-115] minutes. Rescue-PCI was required in 929 patients (34.3%), in whom the fibrinolytic-catheterization time was 7.2 [4.9-11.8] hours, compared to 15.7 [6.8-22,7] hours in those who had successful lytic reperfusion. All cause in-hospital mortality occurred in 151 (5.6%) patients, reinfarction in 47 (1.7%) and ischemic stroke in 33 (1.2%). Major bleeding occurred in 73 (2.7%) patients, including 19 (0.7%) cases of intracranial bleeding. C-statistic confirmed that both scores had high predictive values for in-hospital mortality, demonstrated by TIMI AUC-ROC of 0.80 [0,77-0.84] and GRACE AUC-ROC of 0.86 [0.83-0.89]. CONCLUSION: In a real world registry of a decade-long network for the treatment of ST-elevation myocardial infarction based on the pharmacoinvasive strategy, low rates of in-hospital mortality and cardiovascular outcomes were observed, despite prolonged time metrics for both fibrinolytic therapy and rescue-PCI. Register Clinicaltrials.gov NCT02090712 date of first registration 18/03/2014.

Depression and associated factors among Brazilian adults: the 2019 national healthcare population-based study.

BACKGROUND: Mental disorders represent a major public health challenge worldwide, affecting 80% of people living in low- and middle-income countries. Depression, a mental disorder, is a chronic disease of long duration that causes changes in the brain, resulting from a combination of genetic, physiologic, environmental, and behavioral factors. The aim of this study was to investigate possible factors associated with depression in Brazilian adults. METHODS: A population-based, cross-sectional study was carried out using the public domain database of the 2019 National Health Survey, conducted in Brazil. Depression was considered the dependent variable, and through hierarchical analysis, predictor variables were investigated such as, at the distal level-socioeconomic variables, at the intermediate level-variables related to lifestyle behavior, health condition, and history, and at the proximal level-demographic variables. Logistic regression analysis was used to obtain the adjusted Odds Ratio and the respective 95% confidence interval to identify possible factors associated with depression. RESULTS: The study included 88,531 participant records with 10.27% diagnosed with depression. The adjusted association measurements, after selecting the independent variables in the hierarchical analysis, showed the following factors associated with depression with differing magnitudes: age, brown and white race/skin color, female sex, poor, very poor, or regular self-reported health condition, diagnosis of cardiovascular disease, work-related musculoskeletal disorder, history of smoking habit, and macroeconomic region. CONCLUSIONS: An effective strategy for preventing and managing depression in Brazilian adults must include the control of health status and lifestyle behavior factors, with actions and programs to reduce people's exposure to these factors, understanding that socioeconomic-demographic differences of each population can potentially reduce the disease burden.

Effects of intrauterine exposure to SARS-CoV-2 on infants' development: a rapid review and meta-analysis.

Potential medium- and long-term neurodevelopmental sequelae of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection during pregnancy have not been ruled out. We aimed to systematically review and summarize the evidence regarding the effects of intrauterine exposure to SARS-CoV-2 on infant development and behavior. Scopus, PubMed, Web of Science, CINAHL, and PsycNet databases were searched for studies published up to February 6, 2023, investigating the effects of gestational SARS-CoV-2 on infant development and behavior. We performed narrative synthesis according to updated protocols. Studies using comparison groups and with the Ages and Stages Questionnaires-Third Edition (ASQ-3) scores available were included in a meta-analysis performed according to Cochrane protocols. We used the Newcastle-Ottawa Quality Assessment Scale to analyze the risk of bias. Heterogeneity was calculated using the I2 statistic. The search identified 2,782 studies. After removing duplicates and applying the eligibility criteria, we performed a narrative synthesis of 10 included studies and a meta-analysis of three. There was no evidence of higher developmental delay rates in infants exposed to SARS-CoV-2 during pregnancy compared to non-exposed infants. However, the exposed infants scored lower than either the non-exposed children or pre-pandemic cohorts in some domains. Pooled results from the random-effects model indicated that SARS-CoV-2-exposed infants had lower scores on fine motor (mean difference [MD] = -4.70, 95% confidence interval [CI]: -8.76; -0.63), and problem-solving (MD = -3.05, 95% CI: -5.88; -0.22) domains than non-exposed infants (heterogeneity: I2 = 69% and 88%, respectively). There was no difference between the exposed and non-exposed infants in the communication, gross motor, and personal-social ASQ-3 domains.  Conclusion: We did not find evidence confirming the association between SARS-CoV-2 gestational exposure and neurodevelopmental delays. However, the meta-analysis indicated that gestational exposure negatively affected fine motor and problem-solving skills. Robust evidence on this topic is still incipient, and the available studies present methodological inconsistencies that limit the drawing of clear-cut conclusions.   PROSPERO registration: #CRD42022308002; March 14, 2022. What is Known: • COVID-19 is associated with adverse pregnancy outcomes potentially linked to neurodevelopmental delays. • SARS-CoV-2 vertical transmission is rare; however, infections during pregnancy can be deleterious to the fetus, possibly mediated by maternal immune activation and other inflammatory mechanisms. What is New: • No evidence of increased developmental delay rates among SARS-CoV-2 gestational-exposed infants was found. However, a meta-analysis of three studies showed lower scores in fine motor and personal social ASQ-3 domains among exposed infants. • SARS-CoV-2 gestational exposure and the pandemic can affect child development via many mechanisms. Potential neurodevelopmental sequelae of SARS-CoV-2 exposure during gestation have not been ruled out.

Pre- and perinatal exposures associated with molar incisor hypomineralization: Birth cohort, Brazil.

OBJECTIVE: To analyze prenatal and perinatal stressors associated with molar incisor hypomineralization (MIH) in adolescents. METHODS: Prospective cohort study collected prenatal (socioeconomic status, maternal age, number of prenatal visits, smoking, obesity during pregnancy, abortion history, gestational hypertension) and perinatal stressors (type of delivery, gestational age, birth weight, intensive care unit-ICU at birth). The outcome was MIH at 18-19 years follow-up (n = 590). MIH was defined according to the Ghanim criteria - Model I. We performed a sensitivity analysis, including opacities demarcated in index tooth, incisive or molars, Model II. Through structural equation modeling, we analyzed direct and mediating pathways between multiple stressors with outcomes. RESULTS: MIH was observed in 15.25% (n = 90), and opacities demarcated in any index tooth were observed in 22.8% of adolescents (n = 135). In Model I, no stressor explained MIH significantly, although we watched high standardized coefficients (SC) for low birth weight (SC = 0.223, p = 0.147), lower gestational age (SC = 0.351; p = 0.254), and ICU admission (SC = 0.447, p = 0.254). In Model II, advanced maternal age (SC = 0.148; p < 0.05) and not undergoing prenatal care (SC = 0.384, p < 0.03) explained opacities demarcated in incisors or molars. CONCLUSION: Advanced maternal age and not undergoing prenatal care were associated with MIH lesion-like in incisors or molars.

Intersectoral Collaboration to Promote Child Development: The Contributions of the Actor-Network Theory.

An integrated intersectoral care model promises to meet complex needs to promote early child development and address health determinants and inequities. Nevertheless, there is a lack of understanding of actors' interactions in producing intersectoral collaboration networks. The present study aimed to analyze the intersectoral collaboration in the social protection network involved in promoting early child growth and development in Brazilian municipalities. Underpinned by the tenets of actor-network theory, a case study was conducted with data produced from an educational intervention, entitled "Projeto Nascente." Through document analysis (ecomaps), participant observation (in Projeto Nascente seminars), and interviews (with municipal management representatives), our study explored and captured links among actors; controversies and resolution mechanisms; the presence of mediators and intermediaries; and an alignment of actors, resources, and support. The qualitative analysis of these materials identified three main themes: (1) agency fragility for intersectoral collaboration, (2) attempt to form networks, and (3) incorporation of fields of possibilities. Our findings revealed that intersectoral collaboration for promoting child growth and development is virtually non-existent or fragile, and local potential is missed or underused. These results emphasized the scarcity of action by mediators and intermediaries to promote enrollment processes to intersectoral collaboration. Likewise, existing controversies were not used as a mechanism for triggering changes. Our research supports the need to mobilize actors, resources, management, and communication tools that promote processes of interessement and enrollment in favor of intersectoral collaboration policies and practices for child development.

Investigating oral and systemic pathways between unhealthy and healthy dietary patterns to periodontitis in adolescents: A population-based study.

AIM: To investigate pathways between unhealthy and healthy dietary patterns and periodontitis in adolescents (18-19 years of age). MATERIALS AND METHODS: This population-based study (n = 2515) modelled direct and mediated pathways (via biofilm and obesity) from patterns of healthy diet (fruits, fibre, vegetables, and dairy) and unhealthy diet (sugars, snacks, and salty/fast foods) with initial periodontitis (bleeding on probing [BoP], probing depth [PD] ≥ 4 mm, clinical attachment loss [CAL] ≥ 4 mm), moderate periodontitis (BoP, PD ≥ 5 mm, and CAL ≥ 5 mm), and European Federation of Periodontology and the American Academy of Periodontology (EFP-AAP) periodontitis definitions, adjusting for sex, socio-economic status, smoking, and alcohol, through structural equation modelling (α = 5%). RESULTS: Higher values of healthy diet were associated with lower values of initial periodontitis (standardized coefficient [SC] = -0.160; p < .001), moderate periodontitis (SC = -0.202; p < .001), and EFP-AAP periodontitis (p < .05). A higher value of unhealthy diet was associated with higher values of initial periodontitis (SC = 0.134; p = .005) and moderate periodontitis (SC = 0.180; p < .001). Biofilm mediated the association between higher values of unhealthy diet and all periodontal outcomes (p < .05). CONCLUSIONS: Our findings suggest that both healthy and unhealthy dietary patterns may contribute to reduced or increased extent and severity of periodontitis by local and systemic mechanisms, preceding the effect of other established causes such as smoking and obesity, in younger population.

Sugar intake above international recommendations and oral disease burden: A population-based study.

OBJECTIVE: To evaluate the association between added sugar intake above the daily limit for the risk of noncommunicable diseases (NCDs) and the Chronic Oral Disease Burden in adolescents. METHODS: This was a population-based study using cross-sectional data nested to RPS Cohorts Consortium, São Luís, Brazil, from the 18-19-year-old follow-up (n = 2515). High consumption of added sugars was estimated according to the limits of the World Health Organization guidelines (WHO) (≥5% of total energy/day) and the American Heart Association statement (AHA) (≥25 g/day). The Chronic Oral Disease Burden was a latent variable (number of decayed teeth, periodontal probing depth ≥4 mm, clinical attachment level ≥3 mm, and bleeding on probing). Models were adjusted for Socioeconomic Status, sex, obesity, and plaque index and analyzed through structural equation modeling. RESULTS: Adolescents had high sugar consumption according to the WHO (78.6%) and AHA (81.4%) recommendations. High sugar intake, according to WHO (SC = 0.096; p = 0.007) and AHA (SC = 0.056; p = 0.027), was associated with a heavier Chronic Oral Disease Burden. Even half of the recommended dose was sufficient to affect some oral disease indicators. CONCLUSION: Sugar intake over international statements to prevent NCDs is associated with higher Chronic Oral Disease Burden among adolescents.

Adverse pregnancy outcomes and occlusal traits in the primary dentition: A prospective cohort (BRISA).

OBJECTIVES: This study aimed to analyse the relationship between adverse pregnancy outcomes (APO) and occlusal traits in the primary dentition, checking for different mediation paths. SETTING AND SAMPLE POPULATION: Children evaluated at birth (T1), between 12 and 24 months (T2), and between 24 and 36 months (T3) were included. Two hundred and seventeen children who participated in T1 and T2 were randomly selected to perform the occlusion examination. MATERIALS AND METHODS: This is a prospective cohort study (BRISA). The theoretical model was tested by structural equation modelling (SEM), estimating standardized coefficients (Coeff.) (α = 0.05). The primary exposure was APO-a latent variable manifested from three health problems at birth: low birthweight (LBW), pre-term birth (PTB) and intrauterine growth restriction (IUGR), evaluated in T1. The outcomes were four different occlusal traits assessed in T3: overjet, anterior and posterior crossbite, and crowding. Each outcome's direct and indirect effects were tested, mediated by growth, breathing, breastfeeding, and pacifier use. RESULTS: There was no direct association between APO and any of the outcomes: overjet (Coeff. = -0.163, P = .241), anterior crossbite (Coeff. = -0.696, P = .065), posterior crossbite (Coeff. = -0.087, P = .589) and crowding (Coeff. = 0.400, P = .423). The indirect (total and specifics) effects tested also showed no association (P > .05). However, APO was associated with lower child growth in all models; breastfeeding was associated with higher child growth in all models, and pacifier use was associated with overjet (Coeff. = 0.184, P < .001) and posterior crossbite (Coeff. = 0.373, P = .011). CONCLUSION: APO was not a risk factor for overjet, crossbite and crowding in an early stage of the primary dentition by direct and indirect pathways. However, growth has been lower in children with APO and higher in children breastfed. Also, the harmful effects of using a pacifier in dental occlusion are highlighted.

Maternal care and child physical health: Impact of the exposure to adverse experiences during the first year of life of vulnerable children.

INTRODUCTION: Adverse childhood experiences (ACEs) can negatively affect children's current and future health. OBJECTIVES: This study aims to analyse the impact of ACE on the health of 12-month-old infants assessed by a Physical Health and Maternal Care Indicator (ISCM). METHODS: We conducted a retrospective cohort including 170 infants born in two public services for high-risk births in Brazil. ISCM gathers information that reflects maternal care and the child's health throughout the first year of life, such as vaccination, nutrition, growth, illnesses and accidents. The ACE impact on ISCM was analysed by multiple linear regression, and the d-Cohen test estimated its effect size. Spearman's correlation was used to analyse the cumulative ACE effect, measured by a score reflecting events such as family dysfunction, maternal mental health, poverty and exposure to violence. RESULTS: Most infants were born prematurely (71.7%), had low birthweight (64.7%) and were exposed to three ACEs on average. The ISCM was lower in children exposed to maternal depression (P < 0.001, d-Cohen = 0.08), substance abuse by family members (P = 0.02, d-Cohen = 0.6) and marital conflicts (P = 0.03, d-Cohen = 0.7). The Spearman's correlation showed that the greater the exposure to ACEs, the lower the ISCM (r = -0.40, P < 0.0001). CONCLUSION: Exposure to ACE, especially in the family environment, had negative effect on maternal care and child health. The impact could be detected in the first year of life and had cumulative effect. Our findings indicate the need for a broader approach to child health to minimize ACE's impacts.

[Factors associated with hospitalizations for primary care-sensitive conditions in Brazil: an ecological studyFactores asociados a las hospitalizaciones infantiles por afecciones que podrían tratarse en la atención primaria en Brasil: estudio ecológico]. / Fatores associados às internações infantis por condições sensíveis à atenção primária no Brasil: estudo ecológico.

Objective: To investigate whether structural aspects of primary care units (PCUs) and the work processes of primary care teams are associated with the rate of hospitalizations for primary care-sensitive conditions (HPCSC) in children younger than 5 years of age in Brazil. Method: For this longitudinal ecological study, secondary data were obtained from the Brazilian Hospital Information System and from three cycles of the National Program for Access and Quality Improvement in Primary Care (PMAQ-AB) (2012, 2014, 2017/2018). The analysis included 42 916 PCUs. A multilevel random intercept model with fixed slope was used. In the first level, the outcome (HPCSC rates) and explanatory variables (structure and process indicators) aggregated by PCU were analyzed. Social determinants (represented by a stratification criterion combining municipality population and health care management indicators) were entered in the second level. The t test with Bonferroni correction was used to compare indicator means between regions, and multilevel linear regression was used to estimate the correlation coefficients. Results: The HPCSC rate in children younger than 5 years was 62.78/100 thousand population per estimated PCU coverage area. A direct association with the outcome was observed for: participation in one or more PMAQ-AB cycles; team planning; special hours; dedicated pediatric care area; and availability of vaccines. Equipment, materials, supplies, and being a small or medium-size municipality were inversely associated with HPCSC. Conclusions: HPCSC rates in children below 5 years of age may potentially be reduced through improvements in PCU structure and process indicators and in municipal social determinants.

Objetivo: Determinar la asociación de la estructura de las unidades básicas de salud (UBS) y del proceso de trabajo de los equipos de atención primaria con los determinantes sociales y las tasas de hospitalización por afecciones que podrían tratarse en la atención primaria en menores de 5 años en Brasil. Métodos: Estudio longitudinal ecológico, con datos secundarios del Sistema de Informaciones Hospitalarias y los tres ciclos del Programa Nacional de Mejora del Acceso y de la Calidad de la Atención Básica (PMAQ-AB) (2012, 2014, 2017-2018). Se analizaron 42.916 UBS. Se aplicó un modelo multinivel con pendiente fija e intercepción aleatoria. En el primer nivel, se analizaron el desenlace (tasas de hospitalización por afecciones que podrían tratarse en los servicios de atención primaria) y las variables explicativas (indicadores estructurales y procedimentales) consolidados por UBS. En el segundo nivel, se incluyeron determinantes sociales municipales (representados por un criterio de estratificación que combina el tamaño del municipio con indicadores que influyen en la gestión de salud). Se utilizó la prueba de la t con la corrección de Bonferroni para comparar las medias de los indicadores entre las regiones y la regresión lineal multinivel para estimar los coeficientes de correlación. Resultados: La tasa de hospitalización por afecciones que podrían tratarse en los servicios de atención primaria en menores de 5 años fue de 62,78/100 mil habitantes por área estimada de cobertura de UBS. Los siguientes factores presentaron una asociación directa con el desenlace: participación en uno o más ciclos del PMAQ-AB; planificación del equipo; horario especial; dependencias de atención infantil en la unidad; y disponibilidad de vacunas. La variable relativa a equipos, materiales e insumos y la clasificación como municipio pequeño o mediano se asociaron inversamente con las hospitalizaciones por afecciones que podrían tratarse en los servicios de atención primaria. Conclusiones: Las hospitalizaciones de menores de 5 años por afecciones que podrían tratarse en los servicios de atención primaria pueden reducirse mejorando los indicadores estructurales y procedimentales de las UBS y los determinantes sociales municipales.

Lessons from a Single Amino Acid Substitution: Anticancer and Antibacterial Properties of Two Phospholipase A2-Derived Peptides.

The membrane-active nature of phospholipase A2-derived peptides makes them potential candidates for antineoplastic and antibacterial therapies. Two short 13-mer C-terminal fragments taken from snake venom Lys49-PLA2 toxins (p-AppK and p-Acl), differing by a leucine/phenylalanine substitution, were synthesized and their bioactivity was evaluated. Their capacity to interfere with the survival of Gram-positive and Gram-negative bacteria as well as with solid and liquid tumors was assessed in vitro. Toxicity to red blood cells was investigated via in silico and in vitro techniques. The mode of action was mainly studied by molecular dynamics simulations and membrane permeabilization assays. Briefly, both peptides have dual activity, i.e., they act against both bacteria, including multidrug-resistant strains and tumor cells. All tested bacteria were susceptible to both peptides, Pseudomonas aeruginosa being the most affected. RAMOS, K562, NB4, and CEM cells were the main leukemic targets of the peptides. In general, p-Acl showed more significant activity, suggesting that phenylalanine confers advantages to the antibacterial and antitumor mechanism, particularly for osteosarcoma lines (HOS and MG63). Peptide-based treatment increased the uptake of a DNA-intercalating dye by bacteria, suggesting membrane damage. Indeed, p-AppK and p-Acl did not disrupt erythrocyte membranes, in agreement with in silico predictions. The latter revealed that the peptides deform the membrane and increase its permeability by facilitating solvent penetration. This phenomenon is expected to catalyze the permeation of solutes that otherwise could not cross the hydrophobic membrane core. In conclusion, the present study highlights the role of a single amino acid substitution present in natural sequences towards the development of dual-action agents. In other words, dissecting and fine-tuning biomembrane remodeling proteins, such as snake venom phospholipase A2 isoforms, is again demonstrated as a valuable source of therapeutic peptides.

Minicircle-based expression of vascular endothelial growth factor in mesenchymal stromal cells from diverse human tissues.

BACKGROUND: Mesenchymal stromal cells (MSC) have been exploited for the treatment of ischemic diseases given their angiogenic potential. Despite bone marrow (BM) being the most studied tissue source, cells with similar intrinsic properties can be isolated from adipose tissue (AT) and umbilical cord matrix (UCM). The present study aims to compare the angiogenic potential of MSC obtained from BM, AT and UCM that were genetically modified with vascular endothelial growth factor (VEGF)-encoding minicircle (MC) vectors. The overexpression of VEGF combined with the intrinsic properties of MSC could represent a promising strategy towards angiogenic therapies. METHODS: We established a microporation-based protocol to transfect human MSC using VEGF-encoding MC (MC-VEGF). VEGF production levels were measured by an enzyme-linked immunosorbent assay and a quantitative polymerase chain reaction. The in vitro angiogenic potential of transfected cells was quantified using cell tube formation and migration functional studies. RESULTS: MSC isolated from BM, AT or UCM showed similar levels of VEGF secretion after transfection with MC-VEGF. Those values were significantly higher when compared to non-transfected cells, indicating an effective enhancement of VEGF production. Transfected cells displayed higher in vitro angiogenic potential than non-transfected controls, as demonstrated by functional in vitro assays. No significant differences were observed among cells from different sources. CONCLUSIONS: Minicircles can be successfully used to transiently overexpress VEGF in human MSC, regardless of the cell tissue source, representing an important advantage in a clinical context (i.e., angiogenic therapy) because a standard protocol might be applied to MSC of different tissue sources, which can be differentially selected according to the application (e.g., autologous versus allogeneic settings).

Recombination efficiency measurement by real-time PCR: A strategy to evaluate ParA-mediated minicircle production.

Minicircles (MCs) are DNA molecules that are produced in Escherichia coli by replicating a parental plasmid (PP) and inducing its site-specific intramolecular recombination into miniplasmid (MP; containing the prokaryotic backbone) and MC molecules (comprised by the eukaryotic cassette). The determination of the recombination efficiency and the monitoring of PP, MC and MP species during processing and in the final product are critical aspects of MC manufacturing. This work describes a real-time PCR method for the specific identification of PP, MP or MC that uses sets of primers specific for each species. The method was evaluated using artificial mixtures of (i) PP and MP, (ii) PP and MC and (iii) MP and MC that were probed for all three DNA molecules. The ratio of molecules of each DNA species in these mixtures were determined with differences lower than 10% relatively to the expected ratio of the species in 90% of the mixtures. Next, the recombination efficiency was successfully estimated by analysing pre-purified DNA samples obtained from cell cultures. A standard deviation < 2% was obtained between replicas and results closely correlated with those obtained by densitometry analysis of agarose gels. Further optimization is required to determine recombination efficiency directly from whole cells.

Low bone mineral density is associated with severe periodontitis at the end of the second decade of life: A population-based study.

AIM: To evaluate the association between low bone mineral density (BMD) and severe periodontitis at the end of the second decade of life. MATERIALS AND METHODS: This population-based study analysed 2032 youngers (18-19 years old) of the RPS cohort. BMD of lumbar spine (BMD-LS) and of the whole body (BMD-WB) were assessed by dual x-ray emission densitometry. Low BMD-LS (Z-score ≤ -2) and low BMD-WB (Z-score ≤ -1.5) were correlated with severe periodontitis. The extent of periodontal disease was also evaluated as the following outcomes: proportions of teeth affected by clinical attachment loss ≥5 mm and probing depth ≥5 mm. Multivariate models by sex, education, family income, risk of alcohol dependence, smoking, plaque, bleeding index, and body mass index were estimated through logistic regression (binary outcomes) and Poisson regression (continuous outcomes). RESULTS: The prevalence of severe periodontitis was 10.97%. Low BMD-LS (odds ratio [OR] = 2.08, confidence interval [CI] = 1.12-3.85, p = .01) and low BMD-WB (OR = 1.34, CI = 1.001-1.81, p = .04) were associated with severe periodontitis in the final multivariate models. Low BMD-LS and BMD-WB were also associated with a greater extent of periodontitis (p < .05). CONCLUSIONS: Low BMD was found to be associated with the severity and extent of periodontitis in adolescents. Adolescents at peak bone mass age presenting low BMD are more likely to be affected by severe periodontitis.

Higher sugar intake is associated with periodontal disease in adolescents.

OBJECTIVES: Analyze the association between higher added sugar exposure and periodontal disease in adolescents (18-19 years old). MATERIALS AND METHODS: This was a cross-sectional study nested to RPS Cohorts Consortium, São Luís, Brazil (n = 2515). The exposure was percentage of daily calories from added sugar (≥ 10%), estimated from a quantitative food frequency. The outcome was periodontal disease estimated by the number of teeth affected by bleeding on probing, periodontal probing depth ≥ 4 mm, and clinical attachment level ≥ 4 mm at the same site. A theoretical model was depicted in a directed acyclic graph to identify the minimal sufficient adjustment set: household income, adolescent's educational level, sex, alcohol use, and smoking. Periodontal disease was categorized into < 2 teeth affected, 2 to 3 teeth affected, and ≥ 4 teeth affected to estimate prevalence ratios (PR) by multinomial logistic regression. To test for consistency, means ratio (MR) were estimated using zero-inflated Poisson. RESULTS: High sugar intake was associated with ≥ 4 teeth affected by periodontal disease (PR = 1.42; 95% confidence interval (CI) = 1.03-1.94; p = 0.030); consistency Poisson analysis reinforced these results (MR = 1.15; 95% CI = 1.03-1.29; p = 0.011). CONCLUSION: High level of added sugar intake was associated with greater extent of periodontal disease in adolescents. CLINICAL RELEVANCE: High sugar intake was associated with periodontal disease in adolescents, supporting the integrated hypothesis of dental caries and periodontal disease and giving impetus to future clinical investigation on the effect of restriction of added sugar consumption in periodontal parameters, which potentially may change traditional treatment protocols of periodontal disease.

Optical Gas Sensing with Liquid Crystal Droplets and Convolutional Neural Networks.

Liquid crystal (LC)-based materials are promising platforms to develop rapid, miniaturised and low-cost gas sensor devices. In hybrid gel films containing LC droplets, characteristic optical texture variations are observed due to orientational transitions of LC molecules in the presence of distinct volatile organic compounds (VOC). Here, we investigate the use of deep convolutional neural networks (CNN) as pattern recognition systems to analyse optical textures dynamics in LC droplets exposed to a set of different VOCs. LC droplets responses to VOCs were video recorded under polarised optical microscopy (POM). CNNs were then used to extract features from the responses and, in separate tasks, to recognise and quantify the vapours exposed to the films. The impact of droplet diameter on the results was also analysed. With our classification models, we show that a single individual droplet can recognise 11 VOCs with small structural and functional differences (F1-score above 93%). The optical texture variation pattern of a droplet also reflects VOC concentration changes, as suggested by applying a regression model to acetone at 0.9-4.0% (v/v) (mean absolute errors below 0.25% (v/v)). The CNN-based methodology is thus a promising approach for VOC sensing using responses from individual LC-droplets.

An Overview of Pedagogical Approaches to Caries-Control Medications in Canadian Dental and Dental Hygiene Programs.

OBJECTIVE: To investigate pedagogical approaches and perceived barriers to teaching about caries-control medications, particularly silver diamine fluoride (SDF), in Canadian undergraduate dental and dental hygiene programs. METHODS: In summer 2018, a 9-item questionnaire was distributed to all 10 dental schools and 32 dental hygiene programs in Canada. It enquired about the types of caries-control medications used, teaching methods and perceived barriers to instruction on managing active caries with SDF. METHODS: In summer 2018, a 9-item questionnaire was distributed to all 10 dental schools and 32 dental hygiene programs in Canada. It enquired about the types of caries-control medications used, teaching methods and perceived barriers to instruction on managing active caries with SDF. RESULTS: The response rate was 80% (n = 8) from dental schools and 72% (n = 23) from dental hygiene programs. All curricula included information about conventional caries-control medications: fluoride, silver nitrate and povidone iodine. In all programs, instruction regarding SDF was predominantly didactic: 93% of programs presented lectures on SDF and 30% of programs included clinical teaching and use of SDF in primary dentition only. The lack of consensus on clinical protocols outlining the number and frequency of SDF applications to arrest caries was cited by 43% of the programs as a barrier to clinical teaching. CONCLUSION: There is some variation across Canada in pedagogical approaches to caries-control medications and the inclusion of SDF in curricula. Poorly defined clinical protocols were reported as the main barrier to didactic and clinical use of SDF in undergraduate dental education programs.