Multiple behavioral risk factors for non-communicable diseases among the adolescent population in Brazil: the analysis derived from the Brazilian national survey of school health 2019.

BACKGROUND: Health risk behaviors often emerge or intensify during adolescence and tend to co-occur, exposing individuals to an even greater risk for the development of non-communicable diseases (NCDs). The likelihood of exhibiting multiple health risk factors also increases throughout life and is associated with sociodemographic characteristics contributing to their escalation and severity. In this context, the objective of this study was to analyze the association between sociodemographic characteristics and multiple behavioral risk factors for non-communicable diseases among the adolescent population in Brazil. METHODS: This cross-sectional study utilized data from the Brazilian National Survey of School Health 2019. The sample comprised 121,580 adolescents aged 13 to 17. The analyzed variables included irregular intake of fruits and vegetables, regular consumption of soft drinks and treats, insufficient physical activity, sedentary lifestyle, cigarette smoking, and alcohol consumption. To analyze MBRFs, a classification ranging from zero to five was created, and associations were estimated using Odds Ratio (OR) with the respective 95% confidence interval (CI). The Backward method was employed for the multivariate regression model, utilizing ordinal logistic regression. RESULTS: Adolescents without behavioral risk factors for NCDs constituted only 3.9% (95% CI 3.7-4.1). The most prevalent categories were two and three MBRFs, accounting for 28.3% (95% CI: 27.7-28.8) and 27.0% (95% CI: 26.5-27.5), respectively. Adolescents aged 16 and 17 (ORadj: 1.39; 95% CI: 1.32-1.48), residing in the Brazilian Southeast (ORadj: 1.66; 95% CI: 1.52-1.81), and those reporting poor or very poor self-rated health (ORadj: 2.05; 95% CI: 1.87-2.25) were more likely to exhibit multiple behavioral risk factors. Conversely, male adolescents (adjusted OR: 0.65; 95% CI: 0.62-0.69), those of mixed race (adjusted OR: 0.92; 95% CI: 0.87-0.97), and residents of rural areas (adjusted OR: 0.76; 95% CI: 0.70-0.84) were less likely to manifest MBRFs for NCDs. CONCLUSION: The majority of adolescents displayed MBRFs for NCDs, positively associated with age, region, and perceived health status. This underscores the necessity for healthcare promotional interventions throughout the life cycle, as these behaviors may persist into adulthood.

Evolution of mortality rates among women of reproductive age in Brazilian municipalities: an ecological analysis using Global Burden of Disease estimates.

OBJECTIVES: Women's mortality at a reproductive age has been a global concern, and its decrease has been incorporated as a target of the UN Sustainable Development Goals. The aim of this study was to describe the spatial-temporal evolution of mortality rates among women of reproductive age in Brazilian municipalities by groups of causes and socioeconomic indicators from 2000 to 2018. STUDY DESIGN: Ecological analysis. METHODS: This work was an ecological, descriptive study that analyzed estimates of mortality rates among women of reproductive age (15-49 years) by main groups of causes of death from the Global Burden of Disease (GBD) study in three consecutive trienniums, T1 (2000-2002), T2 (2009-2011), and T3 (2016-2018). To quantify the temporal evolution in mortality rates, the present study calculated the percentage change for each triennium. The spatial analysis of mortality rates was carried out using Moran's index. The Pearson coefficient was used to analyze the correlation between the data. RESULTS: A significant decline in mortality rates was found for all groups of causes in all regions of the country. Despite the downward trend, the percentage change from 2009 to 2011 to 2016 to 2018 showed a decrease in the group of Noncommunicable Diseases (NCDs) and external causes. The decline in mortality rates of women due to external causes showed only a minimal change in the North and Northeast regions from T2 to T3, whereas a cluster of neighboring municipalities with high mortality rates persisted in the municipalities of the South region and in the state of Roraima. The ranking of the main causes of death in Brazilian municipalities showed an increase in neoplasms in detriment to cardiovascular diseases (CVDs). CONCLUSIONS: The main causes of death in women of reproductive age at a more local level could be used to recognize inequalities and to develop interventions aimed at tackling premature and preventable deaths.

Which are the clinical consequences of Molar Incisor hypomineralization (MIH) in children and adolescents? Systematic review and meta-analysis.

OBJECTIVE: This systematic review was conducted to address the following research question: "What are the clinical consequences (outcome) of Molar Incisor Hypomineralization (MIH) (exposure) in children and adolescents (population/patient)?". MATERIAL AND METHODS: After defining the strategy, a search was performed in different databases (MEDLINE via Pubmed, Cochrane Library, BBO, LILACS, Scopus, Web of Science, Embase) and Grey literature in August 2023. Cross-sectional observational studies that identified clinical consequences of MIH (dental caries, post-eruptive structural loss, atypical restorations, hypersensitivity and tooth extraction) were included. The risk of bias was assessed following the Joanna Briggs Institute protocol for cross-sectional studies. Meta-analyses were conducted for each outcome, taking into account the number of patients and teeth. The effect measure considered was the prevalence; random-effects model was adopted. Heterogeneity was assessed using I2 statistics and prediction intervals (PI). RESULTS: A total of 903 studies were identified; 41 were selected for qualitative analysis and 38 for quantitative analysis. Twenty eight studies were classified as presenting uncertain risk of bias, 11 as low risk and 3 as high risk of bias. The prevalence levels, ranked from highest to lowest and considering the tooth and patient units, respectively, were: caries lesions (0.252 - 95% CI 0.158-0.375; 0.512 - 95% CI 0.385-0.639); hypersensitivity (0.286 - 95% CI 0.190-0.407; 0.417 - 95% CI 0.197-0.674), post-eruptive fracture (0.125 - 95% CI 0.099-0.158; 0.257 - 95% CI 0.145-0.412); atypical restorations (0.048 - 95% CI 0.030-0.077; 0.167 - 95% CI 0.096 - 0.274); tooth extraction (0.012 - 95% CI 0.007-0.019; 0.090 - 95% CI 0.019 - 0.331). All meta-analyses resulted in heterogeneity greater than 85%, with the exception of the outcome "tooth extraction" according to the tooth unit (I2 = 57.83). This heterogeneity may be attributed to factors such as differences in the location where the study was realized, the socioeconomic conditions of the studied population, the asymmetric nature of MIH, and patient age. CONCLUSION: The most common consequences of MIH are caries lesions, hypersensitivity, and post-eruptive breakdown. (PROSPERO:CRD42020201410).

Genetic polymorphisms associated with developmental defects of enamel: A systematic review.

BACKGROUND: Polymorphisms in genes related to enamel formation and mineralization may increase the risk of developmental defects of enamel (DDE). AIM: To evaluate the existing literature on genetic polymorphisms associated with DDE. DESIGN: This systematic review was registered in the PROSPERO (CRD42018115270). The literature search was performed in PubMed, Scopus, Web of Science, LILACS, BBO, Cochrane Library, and in the gray literature. Observational studies assessing the association between DDE and genetic polymorphism were included. The Newcastle-Ottawa Scale was used to assess the risk of bias. RESULTS: One thousand one hundred and forty-six articles were identified, and 28 met the inclusion criteria. Five studies presented a low risk of bias. Ninety-two genes related to enamel development, craniofacial patterning morphogenesis, immune response, and hormone transcription/reception were included. Molar-incisor hypomineralization (MIH) and/or hypomineralization of primary second molars (HPSM) were associated with 80 polymorphisms of genes responsible for enamel development, immune response, morphogenesis, and xenobiotic detoxication. A significant association was found between the different clinical manifestations of dental fluorosis (DF) with nine polymorphisms of genes responsible for enamel development, craniofacial development, hormonal transcription/reception, and oxidative stress. Hypoplasia was associated with polymorphisms located in intronic regions. CONCLUSION: MIH, HPSM, DF, and hypoplasia reported as having a complex etiology are significantly associated with genetic polymorphisms of several genes.

Association between dental age and malocclusions: a systematic review.

BACKGROUND: The evidence in the literature suggests that some skeletal or dental malocclusions are involved with dental development, resulting in advanced or delayed dental age (DA). The purpose of this systematic review was to investigate the association between DA and different types of malocclusions. METHODS: The search was carried out on PubMed, Scopus, Web of Science, Virtual Health Library, and in the gray literature. Observational studies that evaluated the association between DA and sagittal, vertical, or transversal malocclusions were included. The quality assessment was performed using the Newcastle-Ottawa Scale (NOS). The data from primary studies were narratively synthesized. The certainty of evidence was evaluated using the GRADE approach. The study was conducted from August 2023 to October 2023. RESULTS: One Thousand Nine Hundred Ninety-One records were identified in the initial search. Twenty (n = 20) studies were included. Most of the studies (n=15) presented a moderate quality according to NOS. Twelve studies evaluated the association between DA and sagittal discrepancies; eight studies evaluated vertical discrepancies, and only one study analyzed a transversal discrepancy. Demirjian's method for DA assessment was the most used among the studies. The primary studies observed that patients of both sexes presenting a vertical growth pattern and males with skeletal Class III malocclusion tend to have advanced DA. The study that investigated transversal malocclusion found that unilateral posterior cross-bite is associated with delayed DA. The certainty of evidence was very low for all outcomes evaluated. CONCLUSION: DA may be associated with the type of malocclusion. It is suggested that DA can be used as an initial diagnostic tool in orthodontics. Future well-designed studies should be performed in order to investigate the association between DA and different types of malocclusions in more detail. TRIAL REGISTRATION: This study was registered in the PROSPERO database (CRD42023454207).

How To Be a Successful Monopartite Begomovirus in a Bipartite-Dominated World: Emergence and Spread of Tomato Mottle Leaf Curl Virus in Brazil.

Begomoviruses are members of the family Geminiviridae, a large and diverse group of plant viruses characterized by a small circular single-stranded DNA genome encapsidated in twinned quasi-icosahedral virions. Cultivated tomato (Solanum lycopersicum L.) is particularly susceptible and is infected by >100 bipartite and monopartite begomoviruses worldwide. In Brazil, 25 tomato-infecting begomoviruses have been described, most of which are bipartite. Tomato mottle leaf curl virus (ToMoLCV) is one of the most important of these and was first described in the late 1990s but has not been fully characterized. Here, we show that ToMoLCV is a monopartite begomovirus with a genomic DNA similar in size and genome organization to those of DNA-A components of New World (NW) begomoviruses. Tomato plants agroinoculated with the cloned ToMoLCV genomic DNA developed typical tomato mottle leaf curl disease symptoms, thereby fulfilling Koch's postulates and confirming the monopartite nature of the ToMoLCV genome. We further show that ToMoLCV is transmitted by whiteflies, but not mechanically. Phylogenetic analyses placed ToMoLCV in a distinct and strongly supported clade with other begomoviruses from northeastern Brazil, designated the ToMoLCV lineage. Genetic analyses of the complete sequences of 87 ToMoLCV isolates revealed substantial genetic diversity, including five strain groups and seven subpopulations, consistent with a long evolutionary history. Phylogeographic models generated with partial or complete sequences predicted that the ToMoLCV emerged in northeastern Brazil >700 years ago, diversifying locally and then spreading widely in the country. Thus, ToMoLCV emerged well before the introduction of MEAM1 whiteflies, suggesting that the evolution of NW monopartite begomoviruses was facilitated by local whitefly populations and the highly susceptible tomato host. IMPORTANCE Worldwide, diseases of tomato caused by whitefly-transmitted geminiviruses (begomoviruses) cause substantial economic losses and a reliance on insecticides for management. Here, we describe the molecular and biological properties of tomato mottle leaf curl virus (ToMoLCV) from Brazil and establish that it is a NW monopartite begomovirus indigenous to northeastern Brazil. This answered a long-standing question regarding the genome of this virus, and it is part of an emerging group of these viruses in Latin America. This appears to be driven by widespread planting of the highly susceptible tomato and by local and exotic whiteflies. Our extensive phylogenetic studies placed ToMoLCV in a distinct strongly supported clade with other begomoviruses from northeastern Brazil and revealed new insights into the origin of Brazilian begomoviruses. The novel phylogeographic analysis indicated that ToMoLCV has had a long evolutionary history, emerging in northeastern Brazil >700 years ago. Finally, the tools used here (agroinoculation system and ToMoLCV-specific PCR test) and information on the biology of the virus (host range and whitefly transmission) will be useful in developing and implementing integrated pest management (IPM) programs targeting ToMoLCV.

Cardiovascular and respiratory evaluation in adenosine A2A receptor knockout mice submitted to short-term sustained hypoxia.

Sustained hypoxia (SH) in mice induces changes in the respiratory pattern and increase in the parasympathetic tone to the heart. Among adenosine G-protein-coupled receptors (GPCRs), the A2A receptors are especially important in mediating adenosine actions during hypoxia due to their expression in neurons involved with the generation and modulation of the autonomic and respiratory functions. Herein, we performed an in vivo evaluation of the baseline cardiovascular and respiratory parameters and their changes in response to SH in knockout mice for A2A receptors (A2A KO). SH produced similar and significant reductions in mean arterial pressure and heart rate in both wild-type (WT) and A2A KO mice when compared to their respective normoxic controls. Mice from WT and A2A KO groups submitted to normoxia or SH presented similar cardiovascular responses to peripheral chemoreflex activation (KCN). Under normoxic conditions A2A KO mice presented a respiratory frequency (fR ) significantly higher in relation to the WT group, which was reduced in response to SH. These data show that the lack of adenosine A2A receptors in mice does not affect the cardiovascular parameters and the autonomic responses to chemoreflex activation in control (normoxia) and SH mice. We conclude that the A2A receptors play a major role in the control of respiratory frequency and in the tachypnoeic response to SH in mice. NEW FINDINGS: What is the central question of this study? Are cardiovascular and respiratory parameters and their changes in response to sustained hypoxia (SH) altered in adenosine A2A receptor knockout mice? What is the main finding and its importance? Cardiovascular parameters and their changes in response to SH were not altered in A2A KO mice. The respiratory frequency in A2A KO was higher than in WT mice. In response to SH the respiratory frequency increased in WT, while it was reduced in A2A KO mice. A2A receptors play a major role in the modulation of respiratory frequency and in the tachypnoeic response to SH in mice.

Oxidation Products from the Neolignan Licarin A by Biomimetic Reactions and Assessment of in vivo Acute Toxicity.

Licarin A, a dihydrobenzofuranic neolignan presents in several medicinal plants and seeds of nutmeg, exhibits strong activity against protozoans responsible for Chagas disease and leishmaniasis. From biomimetic reactions by metalloporphyrin and Jacobsen catalysts, seven products were determined: four isomeric products yielded by epoxidation from licarin A, besides a new product yielded by a vicinal diol, a benzylic aldehyde, and an unsaturated aldehyde in the structure of the licarin A. The incubation with rat and human liver microsomes partially reproduced the biomimetic reactions by the production of the same epoxidized product of m/z 343 [M + H]+. In vivo acute toxicity assays of licarin A suggested liver toxicity based on biomarker enzymatic changes. However, microscopic analysis of tissues sections did not show any tissue damage as indicative of toxicity after 14 days of exposure. New metabolic pathways of the licarin A were identified after in vitro biomimetic oxidation reaction and in vitro metabolism by rat or human liver microsomes.

Tracing the relationship among HIV-1 sub-subtype F1 strains: a phylodynamic perspective.

BACKGROUND: The human immunodeficiency virus type 1, F1 sub-subtype (HIV-1 F1) circulates in three continents: Africa, Europe, and South America. In Brazil, this sub-subtype co-circulates with subtypes B and C and several recombinant forms, mainly BF1 variants. OBJECTIVES: This study aimed to reconstruct the dynamic history of HIV-1 F1 in Brazil. METHODS: HIV-1 near full-length genome and pol gene nucleotide sequences available in public databases were assembled in two datasets (POL671 and NFLG53) to cover the largest number of F1 sub-subtype sequences. Phylodynamic and temporal analyses were performed. FINDINGS: Two main strains of the F1 sub-subtype are circulating worldwide. The first (F1.I) was found among Brazilian samples (75%) and the second (F1.II) among Romanian (62%) and other European and African isolates. The F1 subtype epidemic in Brazil originated from a single entry into the country around 1970. This ancestral sample is related to samples isolated in European countries (France, Finland, and Belgium), which are possibly of African origin. Moreover, further migration (1998 CI: 1994-2003) of strains from Brazil to Europe (Spain and the UK) was observed. Interestingly, all different recombinant BF patterns found, even those from outside Brazil, present the same F1 lineage (F1.I) as an ancestor, which could be related to the acquisition of adaptive advantages for the recombinant progenies. MAIN CONCLUSIONS: These findings are important for the understanding of the origin and dynamics of the F1 sub-subtype and a consequent better and greater understanding of the HIV-1 F1 and BF epidemic that still spreads from Brazil to other countries.

Phenotypic plasticity in Copaifera langsdorffii Desf. in different forest fragments in São Paulo state, Brazil.

Forest fragments are susceptible to environmental shifts and this demands high phenotypic plasticity of the species growing in these areas. In this context, the objective of the present work was to study the phenotypic plasticity of copaíba (Copaifera langsdorffii Desf.) based on morphological and anatomical metrics of the leaflets of plants from six forest fragments. The leaflets of C. langsdorffii individuals of the different fragments did not show qualitative differences, nonetheless, they demonstrated quantitative plasticity. Stomatal density (p = 0.017), specific leaf area (p = 0.009), palisade parenchyma (p = 0.008) and relative water content (p = 0.002), indicated a high luminous, water and nutritional influence on the development of leaflets. Based on the dry mass of the leaflets and the thickness of the palisade parenchyma, the principal component analysis explained 57.43% of the differences found between the variables. The data presented here provides evidence of the phenotypic plasticity of C. langsdorffii which, although occurring in similar soils, showed significant quantitative differences in its morphoanatomical characters.

Improving accuracy of early dental carious lesions detection using deep learning-based automated method.

OBJECTIVE: To investigate the effectiveness of a convolutional neural network (CNN) in detecting healthy teeth and early carious lesions on occlusal surfaces and to assess the applicability of this deep learning algorithm as an auxiliary aid. MATERIALS AND METHODS: A total of 2,481 posterior teeth (2,459 permanent and 22 deciduous teeth) with varying stages of carious lesions were classified according to the International Caries Detection and Assessment System (ICDAS). After clinical evaluation, ICDAS 0 and 2 occlusal surfaces were photographed with a professional digital camera. VGG-19 was chosen as the CNN and the findings were compared with those of a reference examiner to evaluate its detection efficiency. To verify the effectiveness of the CNN as an auxiliary detection aid, three examiners (an undergraduate student (US), a newly graduated dental surgeon (ND), and a specialist in pediatric dentistry (SP) assessed the acquired images (Phase I). In Phase II, the examiners reassessed the same images using the CNN-generated algorithms. RESULTS: The training dataset consisted of 8,749 images, whereas the test dataset included 140 images. VGG-19 achieved an accuracy of 0.879, positive agreement of 0.827, precision of 0.949, negative agreement 0.800, and an F1-score of 0.887. In Phase I, the accuracy rates for examiners US, ND, and SP were 0.543, 0.771, and 0.807, respectively. In Phase II, the accuracy rates improved to 0.679, 0.886, and 0.857 for the respective examiners. The number of correct answers was significantly higher in Phase II than in Phase I for all examiners (McNemar test;P<0.05). CONCLUSIONS: VGG-19 demonstrated satisfactory performance in the detection of early carious lesions, as well as an auxiliary detection aid. CLINICAL RELEVANCE: Automated detection using deep learning algorithms is an important aid in detecting early caries lesions and improves the accuracy of the disease detection, enabling quicker and more reliable clinical decision-making.

Breastfeeding and Cleft Lip and Palate: A Systematic Review and Meta-Analysis.

OBJECTIVE: To evaluate the association between cleft lip and/or cleft palate (CL/P) and breastfeeding (BF). DESIGN: A systematic review and meta-analysis were performed based on studies published in PubMed, Scopus, Web of Science, Cochrane Library, LILACS, BBO, and Embase databases, and in the gray literature. The search occurred in September 2021 and was updated in March 2022. Observational studies evaluating the association between BF and CL/P were included. Risk of bias was analyzed using the Newcastle-Ottawa Scale. A random-effects meta-analysis was conducted. Certainty of evidence was evaluated using the GRADE approach. MAIN OUTCOME MEASURE(S): Frequency of BF in relation to the presence or absence of CL/P, as well as to the type of CL/P. The association between cleft type and BF challenges was also evaluated. RESULTS: From a total of 6863 studies identified, 29 were included in the qualitative review. Risk of bias was moderate and high in most studies (n = 26). There was a significant association between the presence of CL/P and absence of BF (OR = 18.08; 95% CI 7.09-46.09). Individuals with cleft palate with or without cleft lip (CP ± L) had a significantly lower frequency of BF (OR = 5.93; 95% CI 4.30-8.16) and a significantly higher frequency of BF challenges (OR = 13.55; 95% CI 4.91-37.43) compared to individuals with CL. Certainty of the evidence was low or very low in all analyses. CONCLUSION: The presence of clefts, especially those with palate involvement, is associated with higher chances of absence of BF.

Knowledge, attitudes and practices of undergraduate students and dentists about dental prenatal care.

INTRODUCTION: Prenatal care is a fundamental moment for health promotion to be carried out, since at that moment women are more receptive to new knowledge, with the aim of providing better healthcare for the baby. This study aimed to evaluate the knowledge, attitudes and practices of undergraduate students and professionals, both from the public and from private sectors in Brazil, regarding dental prenatal care. MATERIALS AND METHODS: This study corresponded to an online questionnaire survey, via Google Forms, consisting of 17 general questions for undergraduates (n = 103) and 16 for professionals (n = 227) for demographic and school mapping, and 18 specific questions about the importance and protocol of dental prenatal care. T-test, Mann-Whitney U-test and multiple linear regression were adopted (p < .05). RESULTS: The average total score of the questionnaire for students and professionals was 12.40 and 15.65, respectively (p < .0001), indicating moderate knowledge. Professionals showed a higher prevalence of moderate (77%) and high (2%) knowledge of the subject when compared to undergraduate students (51% and 0% respectively). The graduation period in which the students were enrolled was a predictor of the total score of the questionnaire [F (1, 101) = 21.21; p < .0001; R2  = .165]. The female gender and the lower weekly workload were the main predictors of the total score of the questionnaire for professionals [F (3, 223) = 6.74; p < .0001; R2  = .083]. CONCLUSIONS: Although professionals have greater knowledge about dental prenatal care than students, there are still deficiencies in the knowledge and practices of them in respect of dental management during pregnancy. Higher education institutions need to change the teaching-learning plan regarding the holistic dental approach for women during pregnancy.

Angiogenesis in patient-derived xenografts of odontogenic myxoma.

Previously, by employing 3D organotypic tissue culture and patient-derived xenograft (PDX) model, oral myxoma response to a MAPK/MEK inhibitor was observed. Gross examination of the tumour fragments obtained after 55 days of PDX grafting revealed increased capsule vascularization. Microscopic analyses showed blood capillaries intermixed with myxoma cells, but the origin of these capillaries, from mice or humans, was not established. This study aimed to investigate whether the endothelial cells observed in the myxoma PDX model are derived from the mouse or from the primary human tumour. Immunohistochemistry was performed on five tumour fragments from the PDX of myxoma after 55 days of implantation in mice. Immunopositivity for antibodies against human (HLA-ABC) and mouse (H2 Db/H2-D1) major histocompatibility complex class I (MHCI) was assessed in the endothelial cells. The endothelial cells in the PDX fragments revealed a membrane staining for the human MHCI protein in the PDX tumour and adjacent connective tissue capsule, indicating that capillaries were derived from the human tumour fragment. Considering the probable human origin of the endothelial cells from capillary blood vessels in the myxoma PDX, we conclude that this PDX model is an interesting model to study myxoma angiogenesis.

Central giant cell granulomas of the jaws stromal cells harbour mutations and have osteogenic differentiation capacity, in vivo and in vitro.

BACKGROUND: Central giant cell granulomas (CGCG) of the jaws are osteolytic lesions that may behave aggressively and respond poorly to surgery. Microscopically, in addition to giant cells, there is a mononuclear cell population composed of macrophage/monocytic cells and spindle-shaped cells of mesenchymal origin. Seventy two percent of these tumours harbour mutually exclusive TRPV4, KRAS and FGFR1 mutations. We aimed to assess the mutational status of mononuclear and giant cells and the osteogenic potential of stromal cells in vitro and in vivo. METHODS AND RESULTS: We screened CGCG for signature mutations and used laser-capture microdissection to demonstrate that the mutations are restricted to the mononuclear cells. Additionally, we established CGCG primary cell culture and observed that the cells retained the mutations throughout passages. By flow cytometry, we observed predominance of CD14- CD51- CD61- cells, consistent with the expected profile for stromal cells. Considering the mesenchymal origin of stromal cells, we assessed the osteogenic differentiation potential of CGCG cells in culture by cytochemistry (von Kossa and alizarin red staining), alkaline phosphatase (ALP) activity assay and gene expression of osteogenic markers. CGCG cells presented self-capacity to increase ALP levels in a time-dependent manner and under osteogenic induction presented increasing number of calcium deposits, and overall higher expression of osteocalcin, RUNX2, ALPL and osteopontin than cells without osteogenic induction. A patient-derived xenograft model for CGCG was established, and osteoid material deposition was observed. CONCLUSION: Collectively, the results confirm that the signature mutations are restricted to stromal cells in CGCG, and the in vitro and in vivo results support that these cells have the capacity to differentiate into osteoblasts, in line with the bone formation often observed in the stroma of these lesions.

Cytotoxic Cyclotides from Anchietea pyrifolia, a South American Plant Species.

Cyclotides are mini-proteins with potent bioactivities and outstanding potential for agricultural and pharmaceutical applications. More than 450 different plant cyclotides have been isolated from six angiosperm families. In Brazil, studies involving this class of natural products are still scarce, despite its rich floristic diversity. Herein were investigated the cyclotides from Anchietea pyrifolia roots, a South American medicinal plant from the family Violaceae. Fourteen putative cyclotides were annotated by LC-MS. Among these, three new bracelet cyclotides, anpy A-C, and the known cycloviolacins O4 (cyO4) and O17 (cyO17) were sequenced through a combination of chemical and enzymatic reactions followed by MALDI-MS/MS analysis. Their cytotoxic activity was evaluated by a cytotoxicity assay against three human cancer cell lines (colorectal carcinoma cells: HCT 116 and HCT 116 TP53-/- and breast adenocarcinoma, MCF 7). For all assays, the IC50 values of isolated compounds ranged between 0.8 and 7.3 µM. CyO17 was the most potent cyclotide for the colorectal cancer cell lines (IC50, 0.8 and 1.2 µM). Furthermore, the hemolytic activity of anpy A and B, cyO4, and cyO17 was assessed, and the cycloviolacins were the least hemolytic (HD50 > 156 µM). This work sheds light on the cytotoxic effects of the anpy cyclotides against cancer cells. Moreover, this study expands the number of cyclotides obtained to date from Brazilian plant biodiversity and adds one more genus containing these molecules to the list of the Violaceae family.

Food promoted on an online food delivery platform in a Brazilian metropolis during the COVID-19 pandemic: a longitudinal analysis.

OBJECTIVE: To analyse food advertised on an online food delivery (OFD) platform during 16 weeks of the COVID-19 pandemic in Brazil. DESIGN: Longitudinal study. We randomly selected foods advertised on the OFD app's home page, classifying the food items into water; natural juices and smoothies; vegetables; fruits; traditional meals and pasta; ultra-processed beverages; ice cream and candies, and salty packaged snacks; sandwiches; savoury snacks; and pizza. We also registered the marketing strategies used to promote the food items, such as photos, discounts, 'combo deals', and messages on healthiness, value for the money, tastiness, and pleasure. SETTING: Belo Horizonte, Minas Gerais, Brazil. PARTICIPANTS: 1,593 food items. RESULTS: In general, the OFD platform most commonly promoted traditional meals and pasta, ultra-processed beverages, and sandwiches-these food groups were offered 20-25% of the time during the 16 weeks. There were no promotions for water during the whole period, and the least common promotions were those for natural juices and smoothies, vegetables, and fruits (<5%). The most common food promotion strategies were photos (>80%) and discounts (>95%), while approximately 30% of the offers featured combo deals. Messages on tastiness, pleasure, and value for the money varied from 15% to 40%. CONCLUSION: Although the OFD platform offered traditional meals and pasta, most of the foods and beverages advertised were unhealthy and promoted with persuasive strategies. This finding highlights a growing public health concern: an increase in unhealthy eating patterns during the pandemic.

Oxygen-dependent regulation of permeability in low resistance intestinal epithelial cells infected with Giardia lamblia.

Intestinal epithelial cells (IECs) reside in a highly anaerobic environment that is subject to daily fluctuations in partial oxygen pressure (pO2), depending on intestinal tissue perfusion. This condition, known as physiological hypoxia, has a major impact on the maintenance of gut homeostasis, such as effects on the integrity and function of the intestinal epithelial barrier. Giardia lamblia is a microaerophilic protozoan parasite that infects and colonizes the small intestine of its host, causing watery diarrhea. The disease, known as giardiasis, is associated with enhanced intestinal permeability and disruption or reorganization of tight junction (TJ) proteins between IECs. Given the central role of oxygen in gut homeostasis, in this study, we aimed to evaluate whether pO2 affects intestinal permeability (flux of ions and macromolecules) and TJ protein expression in human IECs during G. lamblia infection. Using human cell lines HuTu-80 and Caco-2 as models of "loose" (low resistance) and "tight" (high resistance) intestines, respectively, we elucidated that low pO2 drives intestinal barrier dysfunction in IECs infected with trophozoites through dephosphorylation of protein kinase C (PKC α/ß II). Additionally, we demonstrated that IECs infected with trophozoites in the presence of a pharmacological PKC activator (phorbol 12-myristate 13-acetate) partially restored the barrier function, which was correlated with increased protein expression levels of zonula occludens (ZO)-2 and occludin. Collectively, these results support the emerging theory that molecular oxygen impacts gut homeostasis during Giardia infection via direct host signaling pathways. These findings further our knowledge regarding Giardia-host interactions and the pathophysiological mechanisms of human giardiasis.

Effect of milk replacer feeding rate and frequency of preweaning dairy calves in the southeastern United States: Glucose metabolism.

The objective of this experiment was to examine the effect of milk replacer (MR) feeding rate (FR) and frequency (FF) on glucose metabolism before and after weaning during summer and winter in the subtropical climate of the southeastern United States. Holstein calves (n = 48/season) were enrolled at 8 d of age (DOA) in the summer (June to August, body weight = 40.6 ± 0.7 kg) and winter (November to January, body weight = 41.9 ± 0.8 kg). In each season, calves were randomly assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement including 2 FR [0.65 (low) or 0.76 kg of solids/d (high) of a 26% CP and 17% fat MR] and 2 FF [2× (0700 and 1600 h) or 3× (0700, 1600, and 2200 h)]. Calves were managed similarly and housed in polyethylene hutches bedded with sand. Milk replacer (12.5%) was fed based on treatments until 42 DOA when FR was reduced by half and offered 1×/d (0700 h) for 7 d. Plasma was collected weekly at 1400 h for analyses of glucose and insulin concentrations in all calves. Pre- and postprandial glucose and insulin concentrations of a subset of calves (n = 10/treatment per season) were measured on 20 DOA. A subset of calves (n = 8/treatment per season) was subjected to an intravenous glucose tolerance test (GTT) on 27 and 57 DOA and insulin challenge on 28 and 58 DOA at 1030 h. Average ambient temperature was 26.1 ± 2.2°C in summer and 12.9 ± 5.4°C in winter. During the preweaning period in both seasons, feeding high increased plasma glucose concentrations compared with low, and increasing FF reduced basal insulin concentrations. Compared with 2×, feeding 3× did not affect postprandial glucose but lowered insulin in the summer, whereas in the winter, increased glucose from 30 to 180 min but lowered insulin from 240 to 420 min after MR feeding. Following GTT before weaning in both seasons, 3× reduced insulin increment and area under the curve compared with 2× without affecting glucose disposal. After weaning, treatment did not affect glucose disposal or insulin responses after GTT during winter, but calves fed 3× had faster glucose disposal and stronger insulin responses than 2× during summer. In both summer and winter, preweaned calves fed 3× had greater decrement and area under the curve of plasma glucose after insulin challenge, suggesting enhanced peripheral tissue insulin response compared with 2×. This effect persisted after weaning only during summer. Increasing FR had no effect on metabolic responses in both seasons. In conclusion, increasing MR FF from 2 to 3 times per day reduced insulin secretion but enhanced insulin response on peripheral tissues of preweaned calves regardless of season.

Effects of milk replacer feeding rate and frequency of preweaning dairy calves in the southeastern United States: Performance, abomasal emptying, and nutrient digestibility.

To evaluate the effects of milk replacer (MR) feeding rate (FR) and frequency (FF) on performance, abomasal emptying, and nutrient digestibility in the southeastern United States, Holstein calves (n = 48/season) were enrolled at 8 d of age (DOA) during summer [June to August, body weight (BW; mean ± SD) = 40.71 ± 4.35 kg] and winter (November to January, BW = 42.03 ± 3.83 kg). Within season, calves were randomly assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement including 2 FR [0.65 (low) or 0.76 kg of solid per day (high) of a 26% crude protein and 17% fat MR], and 2 FF [2× (0700 and 1600 h) or 3× (0700, 1600, and 2200 h) daily]. Calves were housed in polyethylene hutches and managed similarly throughout the trial. Milk replacer (12.5% solids) was fed to calves based on their respective treatments until 42 DOA, when MR allowance was reduced by 50% and offered once a day (0700 h) for the following 7 d until weaning. Calves remained on trial until 63 DOA. Calf starter and water were offered ad libitum. Ambient temperature and relative humidity inside and outside hutches were measured hourly. Starter and MR intakes were recorded daily. Respiration rate and rectal temperature were recorded 3 times a week. Structural growth and BW were measured weekly. Acetaminophen (50 mg/kg of BW) mixed with MR was fed to a subset of calves (0700 h, n = 10/treatment per season) on 20 DOA. Plasma was collected at 15, 30, 60, 90, 120, 150, 180, 210, 240, 270, 300, 330, 360, 420, and 480 min after feeding, to analyze acetaminophen. The acetaminophen concentration-time curve was modeled to the first derivative of Siegel's modified power exponential equation, and the time for plasma acetaminophen to reach maximum (Tmax) was calculated to evaluate abomasal emptying rate. During the pre- (14.9-17.9 DOA) and postweaning (51.0-54.0 DOA) periods, a subset (n = 8/treatment per season) of calves was used to determine the apparent digestibility of nutrients, using chromic oxide as the external marker. Feeding 3× reduced preweaning respiration rate during summer and reduced rectal temperature during winter. Increasing FR improved BW gain and structural growth. Feeding more times per day tended to improve growth during winter but not summer. We found no effect of treatment on nutrient digestibility. Increasing FR had no effect on Tmax during winter but tended to delay Tmax of plasma acetaminophen during summer. Regardless of season, increasing FF lowered Tmax of plasma acetaminophen. In conclusion, increasing FF accelerated abomasal emptying and might reduce heat load of preweaning dairy calves but improved growth only during winter. Increased MR allowance improved growth in both seasons but delayed abomasal emptying only under heat stress conditions.