Search

Random amplified polymorphic DNA (RAPD) analysis technique was undertaken in Aedes albopictus populations from three states in Brazil, Rio de Janeiro (RJ), Minas Gerais (MG) and Pernambuco (PE), to estimate the level of genetic variability and levels of genetic exchange between populations. Allele and genotype frequencies were measured on 47 RAPD loci. Average observed heterozigosity (Ho) ranged from 0.282 in MG to 0.355 in Casa Forte (PE) population. Genetic distances estimates indicated that RJ and MG were more genetically similar than populations from PE. Genetic variation observed in local Brazilian populations was attributed to genetic drift associated with restricted gene flow in recently established populations.

The natural vanilla market, which generates millions annually, is predominantly dependent on Vanilla planifolia, a species characterized by low genetic variability and susceptibility to pathogens. There is an increasing demand for natural vanilla, prized for its complex, authentic, and superior quality compared to artificial counterparts. Therefore, there is a necessity for innovative production alternatives to ensure a consistent and stable supply of vanilla flavors. In this context, vanilla crop wild relatives (WRs) emerge as promising natural sources of the spice. However, these novel species must undergo toxicity assessments to evaluate potential risks and ensure safety for consumption. This study aimed to assess the non-mutagenic and non-carcinogenic properties of ethanolic extracts from V. bahiana, V. chamissonis, V. cribbiana, and V. planifolia through integrated metabolomic profiling, in vitro toxicity assays, and in silico analyses. The integrated approach of metabolomics, in vitro assays, and in silico analyses has highlighted the need for further safety assessments of Vanilla cribbiana ethanolic extract. While the extracts of V. bahiana, V. chamissonis, and V. planifolia generally demonstrated non-mutagenic properties in the Ames assay, V. cribbiana exhibited mutagenicity at high concentrations (5000 μg/plate) in the TA98 strain without metabolic activation. This finding, coupled with the dose-dependent cytotoxicity observed in WST-1 (Water Soluble Tetrazolium) assays, a colorimetric method that assesses the viability of cells exposed to a test substance, underscores the importance of concentration in the safety evaluation of these extracts. Kaempferol and pyrogallol, identified with higher intensity in V. cribbiana, are potential candidates for in vitro mutagenicity. Although the results are not conclusive, they suggest the safety of these extracts at low concentrations. This study emphasizes the value of an integrated approach in providing a nuanced understanding of the safety profiles of natural products, advocating for cautious use and further research into V. cribbiana mutagenicity., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In this study, we employ both targeted and untargeted approaches to explore the metabolomic profiles of Vanilla spp., with a particular focus on V. cribbiana (VCR) and its comparison with V. planifolia (VP). We also examine V. bahiana and V. chamissonis using targeted approaches. Through advanced analytical techniques, our untargeted LC-HRMS approach led to the annotation of 60 metabolites, revealing a complex chemical composition with 34 novel compounds in the Vanilla genus in VCR and VP. These findings highlight significant flavoring compounds and lay the foundation for a subsequent quantitative estimation approach. Our targeted analysis, which measured key molecules, underscores VCR's potential in producing vanillin and acetovanillone at levels comparable to the commercially valuable VP and even higher levels of vanillic acid. This research enriches our understanding of flavor composition in vanilla species and emphasizes the importance of exploring wild relatives of vanilla crop for sustainable production and biodiversity conservation., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Andrea Furtado Macedo, Maria Gabriela Bello Koblitz, Rafael Garret and Joana Paula da Silva Oliveira have patent #PCT/BR2022/050468 pending to Federal University of the State of Rio de Janeiro (UNIRIO) & Federal University of Rio de Janeiro (UFRJ). If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

The Vanilla genus is crucial for global production in food, perfume, and pharmaceutical industries. However, exploitation threatens some species, leading to extinction. Traditional communities use vanilla for medicinal purposes, and there are species like Vanilla chamissonis Klotzsch and Vanilla bahiana Hoehne with potential to occupy the market. For this, methanolic extraction of these two mentioned species was conducted alongside Vanilla planifolia . Analyzes of the cell viability, mutagenic and genotoxic potential were performed. In the Ames test, the assays were performed with concentrations from 0.5 and 5000 μg/ml and on five strains. Only Vanilla planifolia exhibited mutagenicity at the highest concentration in the TA98 strain. Viability tests were performed within a dose range of 0.05-5000 µg/ml and 24, 48, and 72-hour exposures. It was possible to observe a reduction in cell viability observed only at the highest concentration, for all three species and both cell types tested. Genotoxicity induction by the extracts was assessed at concentrations from 0.5 to 500 µg/ml through the cytokinesis-block micronucleus assay. No genotoxic damage or reduction in the Nucleus Division Index (NDI). The study found no mutagenicity, cytotoxicity, or genotoxicity in the species tested, indicating potential human use for food or pharmaceutical purposes., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Throughout humanity's development, the medicinal use of plants has evolved as an observational and empirical matter; Yorubá culture uses Kalanchoe sp. as universal antidotes. They are believed to be "ewé òdúndún" or plants of goodness and are traditionally and religiously used in preparations to treat diseases and physiological disorders. Our main goals were to determine the comparative phytochemical composition panel of two Kalanchoe daigremontiana Raym.-Hamet & H. Perrier aqueous extracts through UHPLC-MS metabolomic approaches and highlight the drug-likeness properties of the most abundant compounds in the complex mixture prepared according to its traditional uses. K. daigremontiana leaf aqueous extracts were prepared by cold water maceration (Kd1) or decoction (Kd2). The metabolomics analysis of the extracts was performed through UHPLC-ESI-MS/MS acquisition in positive and negative modes. An in-house database was built with molecules found in plants from the Crassulaceae family and was applied to the annotation step. Our analysis identified 132 compounds, showcasing variations between negative (ESI(-)) and positive (ESI(+)) ionization modes. Notably, flavonoids such as quercetin and kaempferol glycosides, known for their potent antioxidant properties, were abundant in both extracts. Additionally, bufadienolides, associated with diverse biological activities, were present. Both extracts presented similar phytochemical compositions in terms of the number of compounds. Differential analysis revealed significantly higher intensities for specific molecules in both Kd1 and Kd2 extracts. Considering the relative abundance, Kd1 showed a significant amount of Fatty Acids and Steroids concerning Kd2. Additionally, the 15 most abundant compounds were analyzed using in silico pharmacokinetic and toxicity algorithms (SwissADME, pkCSM, and LAZAR) to predict their drug-likeness profile. The fifteen most abundant molecules (Corchorifatty acid F; 9,12,13-TriHOME; Acetylsagittatin A; Apiin; Kaempferol; L-Malic acid; 19-Oxodesacetylcinobufagin; Lucyoside Q; (12 S,13 S)-Epoxylinolenic acid; Isocitric acid; 4''-O-Acetylafzelin; Hellebrigenin; Senedigitalene; Blumenol C and Sativoside R2) had their ADMET in silico profiles predicted using computational tools. Importantly, our in silico assessments pinpointed six promising compounds with favorable absorption, pharmacokinetics, and drug-likeness properties, suggesting their potential as bioactive agents. This finding underscores the significance of these traditional aqueous extracts in folk medicine. In conclusion, our study not only supports the traditional use of Kalanchoe spp. but also sheds light on the safety and pharmacological potential of these extracts. By exploring different extraction methods and employing metabolomics, we have gained valuable insights into the bioactivity and safety of plant extracts, which can guide future research in this field., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Introduction: Risky drinking (RD) is associated with an increased risk of chronic and infectious diseases, injuries, and violence. This study aimed to assess changes in risky drinking (RD) in Brazil after COVID-19 outbreak, both overall and among individuals with self-reported chronic diseases and mental health disorders., Methods: We conducted three independent, anonymous web surveys in Brazil including adult participants: S1 (April/2020, n=19,257), S2 (August/2020, n=1,590), and S3 (January/2021, n=859). Participants were recruited through adapted snowball sampling and sponsored social network advertisements. RD was assessed using the Alcohol Use Disorder Identification Test-Concise, designed to identify individuals at risk of alcohol-related problems. Logistic regression analyses with bootstrapping (B=2,000) were performed, with stratification by sex, age, education, employment, household size, and the presence of chronic and mental health conditions, as well as lifestyle factors, to address sample imbalances., Results: The estimated prevalence of RD was 45.8% [95%CI 45.5, 46.1] in S1, 35.3% [95%CI 34.9, 35.6] in S2, and 33.7% [95%CI 33.3, 34.0] in S3. Participants with chronic diseases consistently presented lower RD prevalence across all three surveys, compared to those without such conditions. Conversely, individuals with mental health disorders presented higher RD prevalence than those without such diagnoses in S1 and S2, but not in S3., Discussion: Despite the decrease in RD prevalence, monitoring of alcohol consumption trends remains essential for shaping effective public health policies. Additionally, the observed variations among individuals reporting chronic and mental health disorders highlight the need for targeted interventions in future crises., Competing Interests: No conflicts of interest declared concerning the publication of this article.

Background: The 2022 mpox outbreak has affected disproportionately people living with HIV (PLWH) and pre-exposure prophylaxis (PrEP) users., Methods: We conducted a cross-sectional study to evaluate factors associated with laboratory diagnosis of mpox among suspected cases, and access differences between PrEP users and PLWH with confirmed diagnostic., Results: 394 mpox suspected cases were analyzed, 309 (78.4%) confirmed. Most patients with mpox were PLWH (54.4%) and 99 (32%) PrEP users. Mpox cases were likely to be between 25 and 39 years old (aOR=2.8; p=0.042), men who have sex with men/bisexual or transgender women (aOR=17.2; p< 0.001) and to have fever (aOR=4.7; p< 0.001), adenomegaly (aOR=7.2; p< 0.001) and multiple vesicular lesions (aOR=4.2; p< 0.001). Comparing PrEP users to PLWH with confirmed mpox, PrEP users had lesions predominantly with exclusive genital involvement (p=0.016); while PLWH had higher extragenital involvement (p=0.018)., Conclusions: PrEP users and PLWHA were the main epidemiological groups in our cohort. Recognizing the differences between vulnerable populations can contribute to the development public policies to control mpox in settings with reduced access to vaccines., Competing Interests: All authors declare no competing interests., (© 2023 The Authors.)

Background Considering the cross-regulation of Th1 and Th2 responses, we hypothesised that atopic diseases (Th2) inhibit the protective Th1 immune response to Mycobacterium leprae and exacerbates leprosy. Objective In this study, we aimed to evaluate the association between leprosy and atopic diseases. Methods To evaluate the association of atopic diseases with leprosy, we conducted a case-control study that included leprosy patients (n = 333) and their household contacts (n = 93). The questionnaire from the International Study of Asthma and Allergies in Childhood, which is validated in several countries for epidemiological diagnosis of atopic diseases, was applied to determine the occurrence of atopic diseases, allergic rhinitis, asthma, and atopic dermatitis among leprosy patients and the household contacts. Results Considering clinical and epidemiological data, among the leprosy group 51.6% (n = 172) were determined to have at least one atopic disease, while atopy was observed less frequently at 40.86% among household contacts (n = 38). When two or more atopic diseases were assessed, the frequency was significantly higher among the leprosy patients than in the household contacts (21.9% vs. 11.8%; P-value = 0.03). Likewise, the frequency of asthma was significantly higher among leprosy patients (21%) than in the household contacts (10.8%; P-value = 0.02). Thus, our analyses revealed an association of atopic diseases with leprosy, with a significant linear increase in the occurrence of leprosy with an increase in the number of atopic diseases (P-value = 0.01). Limitation Due to the difficulties in recruiting household contacts that have prolonged contact with patients, but are not genetically related to the patient, the household contacts group is smaller than the leprosy patient group. Conclusion The data reveal an association between atopic diseases and leprosy outcomes. This knowledge could improve the treatment of leprosy patients with co-incident atopic diseases.

Manganese (Mn) is one of the essential mineral micronutrients most demanded by cacao. Cadmium (Cd) is highly toxic to plants and other living beings. There are indications that Mn can interact with Cd and mitigate its toxicity. The objective of this study was to evaluate the action of Mn on the toxic effect of Cd in young plants of the CCN 51 cacao genotype, subjected to different doses of Mn, Cd, and Mn+Cd in soil, through physiological, biochemical, molecular, and micromorphological and ultrastructural changes. High soil Mn doses favored the maintenance and performance of adequate photosynthetic processes in cacao. However, high doses of Cd and Mn+Cd in soil promoted damage to photosynthesis, alterations in oxidative metabolism, and the uptake, transport, and accumulation of Cd in roots and leaves. In addition, high Cd concentrations in roots and leaf tissues caused irreversible damage to the cell ultrastructure, compromising cell function and leading to programmed cell death. However, there was a mitigation of Cd toxicity when cacao was grown in soils with low Cd doses and in the presence of Mn. Thus, damage to the root and leaf tissues of cacao caused by Cd uptake from contaminated soils can be attenuated or mitigated by the presence of high Mn doses in soil., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: The present study aimed to apply multivariate pattern recognition methods to predict posttraumatic stress symptoms from whole-brain activation patterns during two contexts where the aversiveness of unpleasant pictures was manipulated by the presence or absence of safety cues., Methods: Trauma-exposed participants were presented with neutral and mutilation pictures during functional magnetic resonance imaging (fMRI) collection. Before the presentation of pictures, a text informed the subjects that the pictures were fictitious ("safe context") or real-life scenes ("real context"). We trained machine learning regression models (Gaussian process regression (GPR)) to predict PTSD symptoms in real and safe contexts., Results: The GPR model could predict PTSD symptoms from brain responses to mutilation pictures in the real context but not in the safe context. The brain regions with the highest contribution to the model were the occipito-parietal regions, including the superior parietal gyrus, inferior parietal gyrus, and supramarginal gyrus. Additional analysis showed that GPR regression models accurately predicted clusters of PTSD symptoms, nominal intrusion, avoidance, and alterations in cognition. As expected, we obtained very similar results as those obtained in a model predicting PTSD total symptoms., Conclusion: This study is the first to show that machine learning applied to fMRI data collected in an aversive context can predict not only PTSD total symptoms but also clusters of PTSD symptoms in a more aversive context. Furthermore, this approach was able to identify potential biomarkers for PTSD, especially in occipitoparietal regions., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Cd contamination in cacao beans is one of the major problems faced by cocoa producing countries in Latin America. Cacao scion-rootstock combinations influence the Cd accumulation in the shoot of the plant. The objective of this work was to carry out a comparative analysis between cacao scion rootstock combinations (CCN 51/BN 34, CCN 51/PS 13.19, CCN 51/PH 16 and CCN 51/CCN 51), contrasting for tolerance to cadmium (Cd) toxicity, by means of leaf proteomic profiles, in order to elucidate molecular mechanisms involved in tolerance to Cd toxicity. Cacao scion-rootstock combinations were grown in soil with 150 mg Cd kg -1 soil, together with the control treatment. Leaf samples were collected 96 h after treatments were applied. There were alterations in the leaf proteome of the cacao scion-rootstock combinations, whose molecular responses to Cd toxicity varied depending on the combination. Leaf proteomic analyzes provided important information regarding the molecular mechanisms involved in the tolerance and intolerance of cacao scion-rootstock combinations to Cd toxicity. Enzymatic and non-enzymatic antioxidant systems, efficient for eliminating ROS, especially the expressions of APX and SOD, in addition to the increase in the abundance of metalloproteins, such as ferredoxins, rubredoxin, ALMT, Trx-1 and ABC-transporter were key mechanisms used in the Cd detoxification in cacao scion-rootstock combinations tolerant to Cd toxicity. Carboxylic acid metabolism, glucose activation and signal transduction were also important processes in the responses of cacao scion-rootstock combinations to Cd toxicity. The results confirmed CCN 51/BN 34 as a cacao scion-rootstock combination efficient in tolerance to Cd toxicity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

Nickel is one of the most released trace elements in the environment and in the case of bioaccumulation in foods and beverages derived from cocoa beans can cause risk to human health. It is very important to understand how plants respond to toxic metals and which are the defense strategies they adopt to mitigate their effects. In the present study we used young plants of T. cacao, submitted to increasing Ni doses (0, 100, 200, 300, 400 and 500 mg Ni kg -1 soil) and evaluated them for a period of 30 days. Doses of Ni, from 300 mg of Ni kg -1 onwards in the soil, promoted changes in photosynthetic, antioxidant, osmoregulatory, transcriptomic and translocation levels, evidenced by the increase in the activity of antioxidant enzymes, proline, glycine betaine, upregulation of the metallothionein 2B gene (Mt2b), and lipid peroxidation of the cell membranes. Foliar gas exchange was severely affected at higher doses of Ni. In addition, reduced levels of stomatal conductivity and transpiration rate were observed from 300 mg Ni kg -1 dose onwards in the soil, which consequently affected CO 2 assimilation. Phytostabilization and exclusion mechanisms control the translocation of Ni from the root to the shoot and reduce harmful effects on plant metabolism. Our results highlighted the toxicity of Ni, a trace element often underestimated in T. cacao. In particular, it was noted that doses of 100 and 200 Ni kg -1 soil, although high, do not induce toxicity in T. cacao plants. But Ni toxicity is observed from 300 mg Ni kg -1 soil onwards. This study contributed to the understanding of the harmful effects of higher doses of Ni in cacao plants and the biochemical processes the plant uses to mitigate the effects of this metal., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

The zinc/iron-regulated transporter-like protein (ZIP) gene family first identified in plants is highly distributed in the plant kingdom. This family has previously been reported to transport several essential and non-essential cationic elements, including those toxic to many economically important crops such as cacao ( Theobroma cacao L.). In this article, we present a detailed study on physicochemical properties, evolution, duplication, gene structure, promoter region and TcZIP family three-dimensional protein structure. A total of 11 TcZIP genes have been identified to encode proteins from 309 to 435 aa, with localization in the plasma membrane and chloroplast, containing 6-9 putative domains (TM). Interspecies phylogenetic analysis subdivided the ZIP proteins into four groups. Segmental duplication events significantly contributed to the expansion of TcZIP genes. These genes underwent high pressure of purifying selection. The three-dimensional structure of the proteins showed that α helix conformations are predominant with several pocket sites, containing the metal binding site, with the residues leucine (LEU), alanine (ALA), glycine (GLY), serine (SER), lysine (LYS) and histidine (HIS) the most predicted. Regarding the analysis of the protein-protein interaction and enrichment of the gene ontology, four biological processes were assigned, the most important being the cation transport. These new discoveries expand the knowledge about the function, evolution, protein structures and interaction of ZIP family proteins in cacao and contribute to develop cacao genotypes enriched with important mineral nutrients as well as genotypes that bioaccumulate or exclude toxic metals., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Pacheco, Santana, Pirovani and de Almeida.)

Investigations of the compatibility between cacao genotypes of the population of the Parinari series (Pa), resulting from the reciprocal crossing of Pa 30 × Pa 169 and Pa 121 × Pa 169, allowed the verification of the occurrence of the recessive lethal single character called Luteus-Pa. These genotypes have this gene in heterozygosity, which when intercross or self-fertilize, segregate in a 3:1 ratio. Normal (NS) and mutant (MS) seedlings grow normally and, after a period of approximately 30 days of age, MS leaves begin to show a metallic yellow color, followed by necrotic spots, and death of the entire seedling, approximately 40 days after the emergency. The work evaluate the molecular, biochemical and micromorphological responses in NS and MS, with and without cotyledons, resulting from the crossing of the Pa 30 × Pa 169 cacao genotypes, aiming to elucidate the possible lethal mechanisms of the homozygous recessive Luteus-Pa. The presence of the lethal gene Luteus-Pa in the seedlings of the cacao genotypes of the population of the Parinari (Pa), with and without cotyledons, resulting from the crossing of Pa 30 × Pa 169, in addition to regulating the synthesis of proteins related to the photosynthetic and stress defense processes, promoted an increase in the synthesis of proteins involved in the glycolic pathway, induced oxidative stress, altered the mobilization of cotyledonary reserves, the integrity of cell membranes, leaf micromorphology and induced the death of seedlings, soon after depletion of protein and carbohydrate reserves, especially in the absence of cotyledons., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Masson SAS. All rights reserved.)

Vanilla is a worldwide cherished condiment, and its volatile market is associated with the so-called "vanilla crisis". Even though only two species ( Vanilla planifolia and V. × tahitensis ) are cultivated on a large scale for commercial purposes, the Vanilla genus is comprised of 140 species. The present review article discusses the facets of this crisis, and vanilla crop wild relatives (WRs) are showcased as alternatives to overcome them. Historical, taxonomic, and reproductive biology aspects of the group were covered. Emphasis was given to the metabolic characterization of the vanilla crop WRs, highlighting their main chemical classes and the potential flavor descriptors. Many of these species can produce important flavor compounds such as vanillin, vanillic acid, and acetovanillone, among others. Overall, this review compiles valuable information that can help unravel new chapters of the history of this treasured product by evidencing the biotechnological potential of vanilla crop WRs.

Rationale: Re-exposing an animal to an environment previously paired with an aversive stimulus evokes large alterations in behavioral and cardiovascular parameters. Dorsal hippocampus (dHC) receives important cholinergic inputs from the basal forebrain, and respective acetylcholine (ACh) levels are described to influence defensive behavior. Activation of muscarinic M1 and M3 receptors facilitates autonomic and behavioral responses along threats. Evidence show activation of cholinergic receptors promoting formation of nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) in dHC. Altogether, the action of ACh and NO on conditioned responses appears to converge within dHC., Objectives: As answer about how ACh and NO interact to modulate defensive responses has so far been barely addressed, we aimed to shed additional light on this topic., Methods: Male Wistar rats had guide cannula implanted into the dHC before being submitted to the contextual fear conditioning (3footshocks/085 mA/2 s). A catheter was implanted in the femoral artery the next day for cardiovascular recordings. Drugs were delivered into dHC 10 min before contextual re-exposure, which occurred 48 h after the conditioning procedure., Results: Neostigmine (Neo) amplified the retrieval of conditioned responses. Neo effects (1 nmol) were prevented by the prior infusion of a M1-M3 antagonist (fumarate), a neuronal nitric oxide synthase inhibitor (NPLA), a NO scavenger (cPTIO), a guanylyl cyclase inhibitor (ODQ), and a NMDA antagonist (AP-7). Pretreatment with a selective M1 antagonist (pirenzepine) only prevented the increase in autonomic responses induced by Neo., Conclusion: The results show that modulation in the retrieval of contextual fear responses involves coordination of the dHC M1-M3/NO/cGMP/NMDA pathway., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Re-exposure of rats to a previously fear-conditioned environment arouses great alterations in behavioral and cardiovascular parameters. Pieces of works provide putative evidence for the contribution of the dorsal hippocampus (dHC) to contextual conditioning. dHC gathers massive cholinergic inputs from the basal forebrain, and dHC acetylcholine (ACh) is often described as triggering the retrieval of defensive behavior. ACh acts partially through muscarinic receptors (mAChRs) M1R and M3R subtypes. Hence, activation of mAChRs facilitates autonomic and behavioral responses associated with threats and dangers. Therefore, this study explored the likely involvement of M1R and M3R in rat dHC to establish the behavioral and autonomic changes associated with contextual fear retrieval. Male Wistar rats had stainless steel guide cannula implanted into the dHC before being submitted to contextual fear conditioning (6 footshocks, 1.5 mA, 3 s). A catheter placed within the femoral artery allowed autonomic recordings. A variety of drugs were delivered into the dHC 10 min before contextual re-exposure. The choline reuptake inhibitor hemicholinium induced a decrease of the fear conditioned responses, while did not modify it in non-conditioned animals. The non-selective mAChR antagonist atropine also reduced the fear-conditioned responses, as did the selective M1/M3 mAChRs antagonist fumarate. On the other hand, the M1 selective mAChR antagonist pirenzepine inhibited all the autonomic fear responses without affecting animal freezing. These findings support that cholinergic neurotransmission present in the dHC acts through mAChRs to coordinate the expression of fear evoked by contextual conditioning., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The tolerance of Mentha crispa L. (garden mint) cultivated in cadmium-contaminated oxisol for 120 days was analyzed using plant growth variables such as height, the number of leaves and shoots in different Cd exposure periods, as well as assessing the metal concentration absorbed and accumulated in the plant parts (root, stem, and leaves). The maximum adsorption capacity was estimated at 9220 mg kg -1 and used as a reference to establish the different Cd concentrations to be applied in the soil. M. crispa showed tolerance and revealed a reduction of height, the number of leaves and shoots, root development, and secondary toxicity signs such as chlorosis and leaf wilting. Comparing to the stems and leaves, Cd was retained mainly in the roots. PERMANOVA showed that plant growth variables and Cd concentrations in the plant's part were affected by the Cd exposure time. The canonical discriminant analysis demonstrated height as the most affected variable until 45 growing days, and different responses were observed after 75 days. However, the number of shoots was the variable most affected by higher Cd concentrations. The bioaccumulation and translocation factors for all treatments were lower than one, indicating that M. crispa can be considered as an excluder plant and applied for a phytostabilization strategy., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Lead (Pb) is a highly toxic metal for humans, animals and plants even at low concentrations in the soil. The ingestion of chocolate produced from contaminated beans can contribute to consumer exposure to Pb. While, Mn is an element essential for plants and participates as enzymatic cofactors in several metabolic pathways. The objective of this study was to evaluate the influence of Mn on mitigation of Pb toxicity in seedling of the cacao clonal CCN 51 genotype grown in soils with different doses of Pb, Mn and Mn+Pb, through physiological, biochemical, molecular and nutritional responses. It was found that the seedling of the cacao clonal CCN 51 genotype grown in soils with high Pb, Mn and Mn+Pb contents accumulated these heavy metals in the roots and leaves. Mn doses reduced the Pb uptake by root system and prevented that the Pb accumulated at toxic levels in the roots and leaves of the plants. High doses of Pb applied in soil were highly toxic to the plants, leading, in some cases, them to death. However, no Mn toxicity was observed in cocoa plants, even at high doses in the soil. Uptake of Pb and Mn by the roots and its transport into the aerial part of the plant promoted changes in photosynthesis, leaf gas exchange, respiration, carboxylation and in the instantaneous efficiency of carboxylation, reducing in the treatments with the highest concentrations of Pb, and the emission of chlorophyll fluorescence, affecting the efficiency of photosystem 2 and the production of photoassimilates. Besides that, Pb, Mn and Mn+Pb toxicities activated defense mechanisms in plants that alter the gene expression of met, psbA and psbO, increasing in plants subjected to high concentrations of Pb and the activity of the enzymes involved in the cellular detoxification of excess ROS at the leaf level. In addition, high uptake of Mn by root system was found to reduced Pb uptake in plants grown with Mn+Pb in the soil. Therefore, application of Mn in the soil can be used to mitigate the Pb toxicity in seedling of the cacao clonal CCN 51 genotype grown in contaminated soils.

Strychnos pseudoquina is a plant species whose stem bark is used as bitter tonic beverage. The phytochemical analysis, as well as quantification of phenolic constituents and antioxidant activity of ethanolic extracts from S. pseudoquina stem bark, and leaves were conducted. The extracts were tested for mutagenicity (Ames test) and DNA-damaging activity (Plasmid Cleavage test). Leaves recorded the largest amount of flavonoids. The performed high-performance liquid chromatography (HPLC) showed flavonoids such as isorhamnetin and strychnobiflavone (phytochemical markers of the investigated species) in stem barks, but not in leaves. The proanthocyanidin content and antioxidant activity were significantly higher in stem barks than in leaves. Stem bark and leaf extracts presented mutagenic activity against TA98 and TA100 strains with, and without, metabolic activation (S9). The Plasmid Cleavage test did not indicate DNA-damaging activity. Our results suggest that extracts deriving from S. pseudoquina should be used with extreme caution, mainly the stem bark extract, which is widely used in folk medicine.

Ethnopharmacological Relevance: Extracts of orchids have been traditionally used as human phytotherapeutics. Cyrtopodium flavum, for example, due to the analgesic and anti-inflammatory properties, beside the capacity of heal skin lesions has been focus of research. Also Cyrtopodium glutiniferum, an orchid found in the Brazilian southeastern rainforest, is known to synthesize anti-inflammatory glucomannans in the pseudobulbs, as other potentially therapeutic compounds., Aim of the Study: We have reported the first metabolomic analysis focused on the phenols expression of the neotropical orchid Cyrtopodium glutiniferum Raddi, besides free radical scavenging, anti-inflammatory and antiproliferative activities, and the genotoxicity properties of the aqueous extract., Material and Methods: The metabolomics of C. glutiniferum aqueous extract was performed through UHPLC-MS n acquisition. We have detected the scavenging potential of the extract using DPPH assay. The genotoxic potential was performed by Ames Test (0-5000 μg mL -1 ) and micronucleous assay (0-5000 μg mL -1 ) in RAW264.7 cells. The cytotoxic potential of the extract against RAW264.7 was tested by WST-1 assay (0-500 μg mL -1 ). And after all, the RAW264.7 cells were treated with non-cytotoxic concentrations of C. glutiniferum (0-50 μg mL -1 ) to evaluate the antiproliferative and anti-inflammatory potential, besides the mitochondrial activity., Results: From the 55 molecules identified, 45.5% belonged to the phenolic compounds database from Phenol Explorer, 29% to an in-house Orchidaceae molecules database, and 25.5% to both. Among the identified phenolic compounds, 18 subclasses were discriminated, being phenanthrenes the most abundant. Doses-dependent of C. glutiniferum extracts were able to induce DPPH free radicals scavenging and also to increase TA100 His + revertants, in metabolic environment, showing mutagenicity just in the highest concentration, of 5 mg/plate. On Eukaryotic cell models, the extract also has induced dose-response and time-response cytotoxicity against RAW264.7 macrophages, mainly after 48 h and 72 h, even though the extract has not been able to induce the increase of micronucleated cells and mitotic index alteration on Micronucleus assay. The activation and proliferation of macrophages cultures were downregulated after 24 h and 48 h by the non-cytotoxic concentrations of the extract in a dose-dependent manner., Conclusions: The Cyrtopodium glutiniferum metabolomics, anti-inflammatory and anti-proliferative properties observed in this study suggest a therapeutic efficacy of the orchid extract applied in folk medicine., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Cd is a non-essential metal and highly toxic to plants, animals and humans, even at very low concentrations. Cd has been found in cocoa beans and in their products, as in the case of chocolate. Mn plays an important role in photosynthetic and can interact with Cd and attenuate its toxic effects on plants. The objective of this work was to evaluate the mechanisms of Mn response in the mitigation of Cd toxicity in young plants of the CCN 51 cacao genotype submitted to 0.8 mmol Cd kg -1 , 1.6 mmol Mn kg -1 or the combination of 0.4 mmol Cd kg -1  + 0.8 mmol Mn kg -1 soil, together with the control treatment (without addition of Cd and Mn in soil), by means of analysis of changes in the profile of exclusive proteins (EP) and differentially accumulated proteins (DAP). Leaf and root proteins were extracted and quantified from the different treatments, followed by proteomic analysis. About eight DAP and 38 EP were identified in leaves, whereas in roots 43 DAP and 21 EP were identified. Some important proteins induced in the presence of Cd and repressed in the presence of Cd + Mn or vice versa, were ATPases, isoflavone reductase, proteasome and chaperonin. It was concluded that proteins involved in oxidoreduction and defense and stress response processes, in addition to other processes, were induced in the presence of Cd and repressed in the presence of Cd + Mn. This demonstrated that Mn was able to mitigate the toxic effects of Cd on young plants of the CCN 51 cocoa genotype.

Cacao is a crop of global relevance that faces constant demands for improved bean yield. However, little is known about the genomic regions controlling the crop yield and genes involved in cacao bean filling. Hence, to identify the quantitative trait loci (QTL) associated with cacao yield and bean filling, we performed a QTL mapping in a segregating mapping population comprising 459 trees of a cross between 'TSH 1188' and 'CCN 51'. All variables showed considerable phenotypic variation and had moderate to high heritability values. We identified 24 QTLs using a genetic linkage map that contains 3526 single nucleotide polymorphism (SNP) markers. Haplotype analysis at the significant QTL region on chromosome IV pointed to the alleles from the maternal parent, 'TSH 1188', as the ones that affect the cacao yield components the most. The recombination events identified within these QTL regions allowed us to identify candidate genes that may take part in the different steps of pod growth and bean filling. Such candidate genes seem to play a significant role in the source-to-sink transport of sugars and amino acids, and lipid metabolism, such as fatty acid production. The SNP markers mapped in our study are now being used to select potential high-yielding cacao varieties through marker-assisted selection in our existing cacao-breeding experiments., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© The Author(s) 2020.)

Background: Fatigue is a common symptom in patients with inflammatory bowel diseases (IBD). A translated and culturally adapted, instrument with robust psychometric for measuring fatigue in Brazilian patients with IBD is needed., Objective: To translate and cross-culturally adapt the inflammatory Bowel Disease Fatigue Scale (IBD-F) into Brazilian-Portuguese and to test its measurement properties in Brazilian patients with IBD., Methods: Data from 123 patients with IBD were collected. In addition to IBD-F, the Functional Assessment of Chronic Illness Therapy-Fatigue (FACIT-F) was used. The measurement properties tested were: internal consistency, reproducibility (reliability and agreement), construct validity, internal and external responsiveness, and ceiling and floor effects., Results: The Brazilian-Portuguese version of the IBD-F showed excellent internal consistency (Cronbach's alpha of 0.95), excellent reproducibility (ICC=0.97) and a minimal detectable change of 6.0 points. The construct validity was demonstrated with a good correlation between the IBD-F and FACIT-F (r=- 0.46). Effect sizes used for measuring internal responsiveness were moderate among those with Crohn's (0.66) disease and low in patients with ulcerative colitis (0.24). The Brazilian-Portuguese version of the IBD-F presented with high external responsiveness for Crohn's disease (0.84) and with low external responsiveness for ulcerative colitis (0.33). The area under the curve considered for responsiveness was 0.84. Twenty-five percent of floor effects and no ceiling effect were recorded., Conclusion: The Brazilian-Portuguese version of IBD-F has adequate measurement properties and its use can be recommended in clinical practice and research.

This paper aims to discuss the experience of relatives of children and adolescents with rare diseases as a moral experience. Moral experience is characterized by suffering that is socially interpreted as a catastrophic event, mobilizing resources for signification and meaning that allow the reconstruction of identity, the appreciation of itineraries from a rare diagnosis, as well as the search for peers. Thus, the construction of relationships of recognition, alterity, and belonging is fundamental. From a symbolic interactionist perspective, the results show two significant cores: (1) shock as a surprise in the face of an unexpected diagnosis, leading to the search for peers and promotion of social recognition; (2) the cost involved with the course of a rare disease that implies a care work and the acquisition of associative capital as a possibility of strengthening and building the social capital of health care.

Only a few cultivated species of Vanilla are used to produce vanilla, despite the high demand, predatory exploitation, and low genetic variability that threaten the production of natural vanilla. Vanilla bahiana pods from the Atlantic Forest may be an alternative source of natural vanilla. This study applied bottom-up and shotgun proteomics analysis to identify proteins related to flowering, fruiting, and vanilla-flavor production. Extraction solutions, including Tris-HCl buffer, β-mercaptoethanol and SDS, were assayed. SDS proved to be feasible for extraction of Vanilla fruit proteins and could be an alternative to the phenol method of protein extraction. Progenesis QI for Proteomics (QIP) software loaded with an Orchidaceae database identified 2326 proteins in our samples. Among these, 75 were highlighted as useful for the synthesis of compounds related to vanilla flavor, such as vanillin synthase, which was successfully extracted with 1% SDS, which also improved the variety of the extracted proteins. The proteins identified in V. bahiana pods indicate the enzymatic potential of this species, as further validated by quantifying the vanilla in the samples., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Based on interviews with physiotherapists, we explored the arguments produced by them about their actions and the relationships of a change that has been taking place in the profile of illness in children, where an increased prevalence of complex chronic conditions has been observed. Structured in a qualitative approach, data were generated based on two provocative affirmations that generated arguments by the professionals. Data analysis revealed statements that can be referred to as results of power struggles over the performance in this new field of knowledge. Physiotherapists seek to affirm themselves through their knowledge and by approaching other professional categories, especially Medicine, in order to feel recognized and valued. However, we see that these professionals use a simpler discourse with family and caregivers, creating a bond of trust and identification with them, which facilitates their care and access to children. We point out the need for the hospital environment to be recognized as a place for the action of other categories, besides Medicine and Nursing, and the valuation in pediatric physiotherapy.

The objective of this study was to evaluate Cr toxicity in young plants of the CCN 51 Theobroma cacao genotype at different concentrations of Cr 3+ in the soil (0, 100, 200, 400 and 600 mg kg -1 ) through physiological, ultrastructural, antioxidant and molecular changes. Doses of 400 and 600 mg Cr 3+ kg -1 soil severely affected foliar gas exchange, promoted by damages in photosynthetic machinery evidenced by the decrease in CO 2 fixation. Decreased expression of psbA and psbO genes, changes in enzymatic activity and lipid peroxidation also affected leaf gas exchange. A hormesis effect was observed at 100 mg Cr 3+ kg -1 soil for the photosynthetic activity. As a metal exclusion response, the roots of the cocoa plants immobilized, on average, 75% of the total Cr absorbed. Ultrastructural changes in leaf mesophyll and roots, with destruction of mitochondria, plasmolysis and formation of vesicles, were related to the oxidative stress promoted by excess ROS. The activity of the antioxidant enzymes SOD, APX, GPX and CAT and the amino acid proline coincided with the greater expression of the sod cyt gene demonstrating synchronicity in the elimination of ROS. It was concluded, therefore, that the tolerance of the cocoa plants to the toxicity of Cr 3+ depends on the concentration and time of exposure to the metal. Higher doses of Cr 3+ in the soil promoted irreversible damage to the photosynthetic machinery and the cellular ultrastructure, interfering in the enzymatic and non-enzymatic systems related to oxidative stress and gene expression. However, the low mobility of the metal to the leaf is presented as a strategy of tolerance to Cr 3+ ., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Theobroma cacao is cultivated in the shade, in a so-called 'Cabruca' system, in intercropped with Erithryna or other tree species of economic value, and in full sun as a monoculture in irrigated or chemically-irrigated systems. Since it is a species quite intolerant to wind, it is practically impossible to implant cacao crops under full exposure to the sun, or in areas of frequent winds, without the protection of windbreaks, using arboreal species around the area of culture in the form of box. Wind can cause mechanical stimuli in plants, affecting their growth and development. The objective of this work was to evaluate the photosynthetic changes in mature leaves and the molecular, biochemical and ultrastructural changes in young and mature leaves of the CCN 51 cloned genotype of T. cacao subjected to intermittent (IW) and constant (CW) wind, with velocities of 2.5, 3.5 and 4.5 m s-1, during 3, 6 and 12 h of exposure. It was verified that CW and IW, considering different exposure times, interfered directly in stomatal conductance (gs), transpiration (E) and water use efficiency (WUE), causing a reduction of the photosynthetic rate (A) in mature leaves. In addition, the pulvinus and blade of young and mature leaves, exposed to IW and CW with different exposure times (3 and 12 h), showed marked macroscopic and microscopic mechanical injuries resulting from the constant leaf movement. At both speeds, there was rupture of the cell nuclear membrane in pulvinus and the mesophyll tissues, mainly in the young leaves. On the other hand, in young and mature leaves exposed to CW and IW at different speeds and exposure times, there was lipid peroxidation, increased activity of guaiacol (GPX) and ascorbate (APX) peroxidases in most treatments; and altered expression of transcripts of psba and psbo genes related to the phothosynthetic apparatus and Cu-Zn-sod and per genes related to antioxidative enzymes at the rate of 4.5 m s-1. Younger leaves were more intolerant to mechanical stress caused by the wind, since presented greater macro and microscopic damages and, consequently, greater molecular, biochemical and ultrastructural changes. High wind speeds can seriously compromise the development of young leaves of T. cacao plants and affect their productivity., Competing Interests: The authors have declared that no competing interests exist.

Several studies have identified misconceptions about human papillomavirus (HPV), which can lead to negligent behavior and risk to the health of the population. This article aims to assess the knowledge of men and women about HPV in the city of Ipatinga, Minas Gerais, and evaluate socioeconomic factors and preventive measures associated with such knowledge. It involved a cross-sectional study with 591 subjects living in the city, by means of stratified sampling by quotas proportional to the number of individuals by sex and age in each of the eight administrative regions of the city. Pearson's chi-square test or the t-Student test and the binary logistic regression model were used to assess the factors associated with knowledge about HPV. Less than half (40.1%) of the respondents said they knew what HPV was. Factors associated with having knowledge about HPV were being female, having secondary or tertiary education, using the private health service, having heard about or seen a campaign about HPV and knowing of the existence of a vaccine against HPV. There is a great deficit of knowledge about HPV and vague information of what is known, favoring actions with a potential risk to personal health, including that of the partner.

Spontaneous heterotopic pregnancy is a potentially life-threatening condition rarely considered when a patient with an intrauterine pregnancy is asymptomatic or presents with complaints such as abdominal pain. An advanced abdominal pregnancy as the form of the ectopic component, outside the context of assisted reproduction, is an even more unusual and perilous diagnosis with very few cases reported in the literature. We report such a case in a 34-year-old multipara, with no known risk factors for ectopic pregnancy. The pregnancy was initially misdiagnosed and managed as a dichorionic gestation, complicated with fetal demise of one of the twins. The correct diagnosis was only made after term delivery of the intrauterine pregnancy. The abdominal pregnancy was subsequently managed via laparotomy with no complications., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Ltd (unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.)

Selecting parents and evaluating progenies is a very important step in breeding programs and involves approaches such as understanding the initial stages of growth and characterizing the variability among genotypes for different parameters, such as physiological, growth, biomass partitioning and nutrient translocation to the aerial part. In these cases, facilitating tools can be used to understand the involved gene dynamics, such as diallel crosses and genetic and phenotypic correlations. Our main hypothesis is that the contrasting phenotypes of these parental genotypes of cocoa used are due to genetic factors, and progenies derived from crosses of these parental genotypes are useful for breeding programs related to plant architecture, physiological parameters and translocation of mineral nutrients. We aimed to evaluate the combining abilities in progenies of cacao (Theobroma cacao L) originating from contrasting parents for canopy vigor. Emphasis was given to the evaluation of morphological and physiological parameters and the phenotypic and genotypic correlations to understand the dynamics of the action of the genes involved, as well as in expression profile from genes of gibberellins biosynthesis pathway in the parents. Fifteen F1 progenies were obtained from crosses of six clones (IMC 67, P4B, PUCALA, SCA 6, SCA 24 and SJ 02) that were evaluated in a randomized complete block design with four replicates of 12 plants per progeny, in a balanced half table diallel scheme. It is possible to identify and select plants and progenies of low, medium and high height, as there is expressive genetic variability for the evaluated parameters, some of these on higher additive effects, others on larger nonadditive effects and others under a balance of these effects. Most physiological parameters evaluated show that for selection of plants with the desired performance, no complex breeding methods would be necessary due to the high and medium heritability observed. Strong genetic components were observed from many of the correlations, which indicate the possibility to formulate selection indices for multi-traits, such as dwarfism or semidwarfism, tolerance to increase of leaf sodium concentrations and maintenance of the photosynthetic apparatus integrity under these conditions. Additionally, plants with higher carbon fixation, better water use, higher carboxylation efficiency and greater magnesium accumulation in leaves can be selected.

Cadmium (Cd) is a metal known for its genotoxicity and cytotoxicity, much concerned for its potential environmental and human health impacts. This study evaluates the toxic effect of Cd in Calophyllum brasiliense plants. The plants were cultivated for 30 days in full nutrient solution in order to adapt, and for 15 days in nutrient solution without Cd or with 4, 8, 16, and 32 μmol Cd L -1 . Anatomical analysis of the leaf showed no significant effects of Cd on epidermal thickness in abaxial and adaxial sides, palisade, and spongy parenchyma. Contrastingly, changes were noticed in the ultrastructural level in the leaf mesophyll cells as rupture of the membrane of chloroplasts and disorganization of the thylakoid membranes, in starch grains and in mitochondria with rupture of the membrane and invagination of the nuclear membrane. Electron dense materials into cells of the cortex and vascular bundle were also observed. In the cells of the root system, the observed ultrastructural changes were disruption of the cell wall and electron dense material deposition in the cortex cells and vascular region. Cd accumulated in roots with low translocation into shoot. Cd toxicity also affected the photosynthetic activity, inducing stomatal closure and photosynthetic assimilation reduction and the instantaneous carboxylation efficiency, drastically reducing the leaf transpiration. The nutrient content in the stem and root was variable, according to Cd increase in nutrient solution. Based on the experimental evidence, it can be concluded that C. brasiliense has potential to bioconcentrate high Cd levels in the root system.

The incorporation of computer-aided design/computer-aided manufacturing (CAD/CAM) technology into complete denture fabrication brings about several advantages to the fabrication process, providing better predictability of the desired outcomes and high accuracy of denture fit, mainly because the milling of prepolymerized acrylic resin eliminates the shrinkage of the acrylic base. Also, there is a decrease in the porosity when compared to a conventionally processed denture, and consequently there is a decrease in the retention of Candida albicans on the denture base. The presented workflow for complete denture fabrication presents a totally wax-free manufacturing process, combining rapid prototyping (RP) and rapid milling. With the presented technique, the maxillomandibular relation (MMR) and the ideal setup of the tooth arrangement are developed by using occlusion rims and trial setup made with RP. For the definitive final denture, the denture base and the basal surfaces of the conventional denture teeth were milled according to the individual clinical situation. Posteriorly, the teeth were adapted and bonded into the milled sockets of the milled base., Competing Interests: The authors declare that there are no competing interests regarding the publication of this paper.

Objective: To evaluate dimensional alterations of the alveolar ridge that occurred following tooth extraction at sites grafted with Bio-Oss(®) Collagen., Material and Methods: Twenty-eight subjects with maxillary incisors, canines, and premolars scheduled for extraction were included. The tooth was carefully removed. The patients were randomly assigned to a test or a control group. In the test group patients, Bio-Oss(®) Collagen was placed in the fresh extraction socket while in the controls no grafting was performed. Radiographic examination (cone beam computed tomograms, CBCT) was performed immediately after tooth extraction and socket treatment. Four months later, a new CBCT was obtained. In the radiographs, (i) the distance (mm) between base of the alveolar process (apex) and the buccal and palatal crests was determined, (ii) the outer profile of alveolar process of the experimental sites was outlined, and the cross section of the area (mm(2) ) determined., Results: After 4 months of healing, the buccal and to a less extent also the palatal bone plate had become markedly reduced in height. The placement of a biomaterial in the socket failed to prevent resorption of the buccal and palatal bone walls. The cross-sectional area of the control ridge was reduced about 25% and of the test ridge with 3%., Conclusion: The placement of a xenograft in fresh extraction sockets markedly counteracted the reduction in the hard tissue component of the edentulous sites., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Aedes aegypti has developed evolution-driven adaptations for surviving in the domestic human habitat. Several trap models have been designed considering these strategies and tested for monitoring this efficient vector of Dengue. Here, we report a real-scale evaluation of a system for monitoring and controlling mosquito populations based on egg sampling coupled with geographic information systems technology. The SMCP-Aedes, a system based on open technology and open data standards, was set up from March/2008 to October/2011 as a pilot trial in two sites of Pernambuco -Brazil: Ipojuca (10,000 residents) and Santa Cruz (83,000), in a joint effort of health authorities and staff, and a network of scientists providing scientific support. A widespread infestation by Aedes was found in both sites in 2008-2009, with 96.8%-100% trap positivity. Egg densities were markedly higher in SCC than in Ipojuca. A 90% decrease in egg density was recorded in SCC after two years of sustained control pressure imposed by suppression of >7,500,000 eggs and >3,200 adults, plus larval control by adding fishes to cisterns. In Ipojuca, 1.1 million mosquito eggs were suppressed and a 77% reduction in egg density was achieved. This study aimed at assessing the applicability of a system using GIS and spatial statistic analysis tools for quantitative assessment of mosquito populations. It also provided useful information on the requirements for reducing well-established mosquito populations. Results from two cities led us to conclude that the success in markedly reducing an Aedes population required the appropriate choice of control measures for sustained mass elimination guided by a user-friendly mosquito surveillance system. The system was able to support interventional decisions and to assess the program's success. Additionally, it created a stimulating environment for health staff and residents, which had a positive impact on their commitment to the dengue control program.

Hesperidin (hesperitin-7-O-rutinoside) and hesperitin (hesperitin-7-O-glucoside) show anti-inflammatory, antimicrobial, antioxidant, and anticarcinogenic effects and prevent bone loss. However, hesperidin has a low bioavailability compared to hesperitin due to the rutinoside moiety attached to the flavonoid. The removal of the rhamnose group to yield the corresponding flavonoid glucoside (hesperetin-7-glucoside) improved the bioavailability of the aglycone, hesperetin, in humans. In line with these assumptions, the aim of this work was the enzymatic production of hesperitin from hesperidin with hesperidinase. Despite the low hesperidin solubility in the reaction medium, the enzymatic bioconversion was carried with hesperidin soluble at lower concentrations (≤0.05 mg ml(-1)) and insoluble for high concentrations (>0.1-50 mg ml(-1)). A twofold increase in maximum reaction rates overtook the expected values, pointing to the enzyme ability to degrade insoluble hesperidin. To improve the bioprocess, hesperidinase was tested soluble and immobilized in calcium alginate (2%), k-carrageenan (2%), and chitosan (2%) beads. The immobilization was carried out by adsorption and encapsulation. Chitosan was cross-linked with glutaraldehyde (1% and 2%) and sodium sulfate (13.5% and 15%) in acetate buffer (0.02 M, pH 4.0). The relation between bioprocessing conditions and hesperidinase stability was studied. A residual activity of 193% was obtained with immobilized hesperidinase compared to the soluble form. A half-life of 770 min was attained with hesperidinase encapsulated in calcium alginate beads. The results presented in this work highlight the potential of hesperidinase encapsulation towards hesperitin production with insoluble substrate. To our knowledge, this work presents for the first time the potential of hesperidinase encapsulation on hydrogels for hesperitin production. This is an important achievement for pharmaceutical and nutraceutical applications of hesperitin because this compound presents a higher bioavailability compared to hesperidin., (Copyright © 2012 John Wiley & Sons, Ltd.)

Coutarea hexandra is a species commonly known in Brazil as quina, and its bark is used in folk medicine. In this study, we assess the mutagenic and DNA-damaging effects of ethanol extracts from C. hexandra stem bark (SCH) and leaves (LCH) by employing the Ames test on the TA98 and TA100 strains of Salmonella typhimurium in addition to a plasmid treatment test. Furthermore, we performed a phytochemical analysis by TLC and HPLC, a quantification of the phenolic constituents and an assessment of the antioxidative activity. SCH and LCH showed mutagenic action in the Ames test for TA98 strains after metabolic activation. LCH also showed mutagenicity for the TA100 strain after metabolic activation. The findings from the plasmid treatment test did not indicate any DNA-damaging activity for either of the extracts with the tested dosages. SCH showed greater flavonoid content and greater antioxidative potential in relation to LCH. This study suggests that caution is advisable in the use of this plant. However, in vivo studies should be conducted to confirm these data., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Objective: : The purpose of this study was to assess the effect of cervical conization on subsequent pregnancy and delivery outcomes., Materials and Methods: : A retrospective case-control study was performed in patients who underwent conization from 2000 to 2005 and had a subsequent delivery. Two case controls were matched for each case studied. The obstetric outcomes of the 2 groups were compared. A subsequent analysis comparing the 2 excisional techniques used was performed., Results: : The study population group included 87 women (29 cases and 58 controls). Overall, the mean gestation age at delivery (38 vs 39 wk, p = .003), prevalence of preterm birth (20.7% vs 5.2%, p = .025), mean birth weight (3,035 vs 3279 g, p = .018), and low birth weight (20.7% vs 1.7%, p = .02) were statistically different among the cases and controls.In the study group (n = 29), large loop excision of the transformation zone was used in 62% (n = 18) and laser was used in 38% (n = 11) of the cases. There was no significant difference in the medium depth of the excised tissue, the prevalence of preterm birth, low birth weight, and the cesarean delivery rate between the 2 techniques., Conclusions: : Despite the small number of cases, this study indicates that excision of the transformation zone is associated with an increased risk of overall preterm delivery and low-birth weight infants in subsequent pregnancies. No significant difference was found between the 2 cervical excision procedures.

The Amyotrophic lateral sclerosis (ALS) is the most common form of motor neuron disease in the adulthood, and it is characterized by rapid and progressive compromise of the upper and lower motor neurons. The majority of the cases of ALS are classified as sporadic and, until now, a specific cause for these cases still is unknown. To present the different hypotheses on the etiology of ALS. It was carried out a search in the databases: Bireme, Scielo and Pubmed, in the period of 1987 to 2011, using the following keywords: Amyotrophic lateral sclerosis, motor neuron disease, etiology, causes and epidemiology and its similar in Portuguese and Spanish. It did not have consensus as regards the etiology of ALS. Researches demonstrates evidences as regards intoxication by heavy metals, environmental and occupational causes, genetic mutations (superoxide dismutase 1), certain viral infections and the accomplishment of vigorous physical activity for the development of the disease. There is still no consensus regarding the involved factors in the etiology of ALS. In this way, new research about these etiologies are necessary, for a better approach of the patients, promoting preventive programs for the disease and improving the quality of life of the patients.

Objective: In this study, we aimed to quantify posture and body image in patients with major depressive disorder during episodes and after drug treatment, comparing the results with those obtained for healthy volunteers., Method: Over a 10-week period, we evaluated 34 individuals with depression and 37 healthy volunteers. Posture was assessed based on digital photos of the subjects; CorelDRAW software guidelines and body landmarks were employed. Body image was evaluated using the Body Shape Questionnaire., Results: During depressive episodes (in comparison with the post-treatment period), patients showed increased head flexion (p<0.001), increased thoracic kyphosis (p<0.001), a trend toward left pelvic retroversion (p=0.012) and abduction of the left scapula (p=0.046). During remission, patient posture was similar to that of the controls. At week 1 (during the episode), there were significant differences between the patients and the controls in terms of head flexion (p<0.001) and thoracic kyphosis (p<0.001); at weeks 8-10 (after treatment), such differences were seen only for shoulder position. The mean score on the Body Shape Questionnaire was 90.03 during the depressive episode, compared with 75.82 during remission (p=0.012) and 62.57 for the controls., Conclusion: During episodes of depression, individuals with major depressive disorder experience changes in posture and mild dissatisfaction with body image. The findings demonstrate that the negative impact of depression includes emotional and physical factors.

Introduction: This study analyses the cases of all bites (including dry bites) caused by Bothropoides jararaca attended at the Vital Brazil Hospital of the Butantan Institute, State of São Paulo, Brazil., Methods: A retrospective study was conducted of patients bitten by Bothropoides jararaca (n=792) from January 1990 to December 2004. The characteristics of the snake specimen, data related to the accident and clinical manifestations on admission were obtained from patient medical records., Results: The majority of the cases in this study were caused by female and juvenile snakes. No stomach contents were found in 93.4% of the snake specimens after dissection. No statistical difference was observed between the occurrence of dry bites and the maturity or sex of the snake. The median SVL of snakes in mild and moderate cases was 40.5 cm and in severe cases, SVL increased to 99 cm. Necrosis was more common in the digits of the feet and hands (4.8%) compared to the other body regions (1.8%). A significant difference was verified between severity and a time interval greater than six hours from the bite to hospital admission. A significant association was verified between gingival bleeding and abnormal blood coagulability. In accidents caused by adult snakes, necrosis was more frequent (7.2%) compared to accidents caused by juvenile snakes (1%)., Conclusions: In this work, the association between certain epidemiological data and the evolution of biological parameters in the clinical course of Bothrops sensu latu accidents were highlighted, contributing to the improvement of snake bite assistance.

The authors report a clinical case of warfarin hypersensitivity noticed when a patient, with thromboembolic risk, was submitted to warfarin. She was found to be heterozigotic to -1639G>A (gene VKORC1) and homozygotic to 1075A>C (genotype CYP2C9*3/*3). In this article the warfarin pharmacodinamics and its role over Vitamin K is revisited. It includes some suggested indications for investigating genetic polymorphisms, in order to avoid complications associated with warfarin.

Background & Objectives: Dengue is currently one of the most important arthropod-borne diseases and may be caused by four different dengue virus serotypes (DENV-1 to DENV-4), transmitted mainly by Aedes aegypti (Diptera: Culicidae) mosquitoes. With the lack of a dengue vaccine, vector control strategies constitute a crucial mode to prevent or reduce disease transmission. In this context, DENV detection in natural Ae. aegypti populations may serve as a potential additional tool for early prediction systems of dengue outbreaks, leading to an intensification of vector control measures, aimed at reducing disease transmission. In Brazil, this type of surveillance has been performed sporadically by a few groups and has not been incorporated as a routine activity in control programs. This study aimed at detecting DENV in natural Ae. aegypti from Recife, Pernambuco, to check the circulating serotypes and the occurrence of transovarial transmission in local mosquito populations., Methods: From January 2005 to June 2006, mosquitoes (adults and eggs) were collected in houses where people with clinical suspicion of dengue infection lived at. RNA was extracted from pooled mosquitoes and RT-PCR was performed in these samples for detection of the four DENV serotypes., Results & Conclusion: Out of 83 pools of adult mosquitoes collected in the field, nine were positive for DENV: five for DENV-1, two for DENV-2 and two for DENV-3. From 139 pools of adult mosquitoes reared from collected eggs, there were 17 positive pools: three for DENV-1, 10 for DENV-2, and four for DENV-3. These results are discussed in the paper in regard to the local dengue epidemiological data. The conclusions clearly point to the informative power and sensitivity of DENV entomological surveillance and to the importance of including mosquito immature forms in this strategy.

Insecticide resistance is one of the main problems in vector control programs. Because insects have developed resistance to all classes of available chemical insecticides, a proper surveillance and management of resistance in areas where these compounds are being utilized is crucial for the success of control programs. Since the mechanisms and molecular bases of resistance are various, they must be characterized to allow efficient monitoring strategies. Here we report the establishment of an Aedes aegypti strain resistant to temephos, named RecR, selected under laboratory conditions. The parental A. aegypti population was obtained from eggs collected in an area where temephos had been used for 8 years, and presented a baseline resistance ratio (RR) of 7. After 17 generations under selective pressure, the RR has increased to 180. Biochemical assays indicate that metabolic mechanisms are involved on temephos resistance in the selected strain. These experiments showed that, compared to the susceptible colony Rockefeller, RecR present higher activity of glutathione S-transferases (GSTs), alpha- and beta-esterases, and, to a lesser degree, mixed function oxidases (MFO). At the 14th or 17th generations, there was no cross resistance of these insects to deltamethrin, cypermethrin and malathion, while a low resistance level (RR=3) was observed for pyriproxyfen, a juvenile hormone analogue. Experiments on resistance reversal, performed through three different field simulated schemes using the resistant strain, showed that temephos susceptibility can be recovered. The establishment of an A. aegypti colony resistant to temephos is extremely valuable for a deeper understanding of resistance mechanisms and thus for further improvements in control strategies against this vector. With the urgent need on improving methodologies to monitor resistance, molecular studies such as microarrays, and resistant colonies such as RecR will certainly hasten such studies., (Copyright 2009 Elsevier B.V. All rights reserved.)

Aedes aegypti is a very efficient disseminator of human pathogens. This condition is the result of evolutionary adaptations to frequent haematophagy, as well as to the colonization of countless types of habitats associated with environmental and cultural factors that favor the proliferation of this mosquito in urban ecosystems. Studies using sensitive methods of monitoring demonstrate that the methods of surveillance used in the Brazilian program do not show the high degrees of the infestation of cities by this vector. To increase the capacity of the health sector, new tools are needed to the practice of surveillance, which incorporate aspects of the vector, place and human population. We describe here the SMCP-Aedes - Monitoring System and Population Control of Aedes aegypti, aiming to provide an entomological surveillance framework as a basis for epidemiological surveillance of dengue. The SMCP-Aedes is uphold in the space technology information, supported by the intensive use of the web and free software to collect, store, analyze and disseminate information on the spatial-temporal distribution of the estimated density for the population of Aedes, based on data systematically collected with the use of ovitraps. Planned control interventions, intensified where and when indicated by the entomological surveillance, are agreed with the communities, relying on the permanent social mobilization.

The Cry48Aa/Cry49Aa mosquitocidal two-component toxin was recently characterized from Bacillus sphaericus strain IAB59 and is uniquely composed of a three-domain Cry protein toxin (Cry48Aa) and a binary (Bin) toxin-like protein (Cry49Aa). Its mode of action has not been elucidated, but a remarkable feature of this protein is the high toxicity against species from the Culex complex, besides its capacity to overcome Culex resistance to the Bin toxin, the major insecticidal factor in B. sphaericus-based larvicides. The goal of this work was to investigate the ultrastructural effects of Cry48Aa/Cry49Aa on midgut cells of Bin-toxin-susceptible and -resistant Culex quinquefasciatus larvae. The major cytopathological effects observed after Cry48Aa/Cry49Aa treatment were intense mitochondrial vacuolation, breakdown of endoplasmic reticulum, production of cytoplasmic vacuoles, and microvillus disruption. These effects were similar in Bin-toxin-susceptible and -resistant larvae and demonstrated that Cry48Aa/Cry49Aa toxin interacts with and displays toxic effects on cells lacking receptors for the Bin toxin, while B. sphaericus IAB59-resistant larvae did not show mortality after treatment with Cry48Aa/Cry49Aa toxin. The cytopathological alterations in Bin-toxin-resistant larvae provoked by Cry48Aa/Cry49Aa treatment were similar to those observed when larvae were exposed to a synergistic mixture of Bin/Cry11Aa toxins. Such effects seemed to result from a combined action of Cry-like and Bin-like toxins. The complex effects caused by Cry48Aa/Cry49Aa provide evidence for the potential of these toxins as active ingredients of a new generation of biolarvicides that conjugate insecticidal factors with distinct sites of action, in order to manage mosquito resistance.