Serotype-Specific Epidemiological Patterns of Inapparent versus Symptomatic Primary Dengue Virus Infections: A 17-year cohort study in Nicaragua.

Dengue is the most prevalent mosquito-borne viral disease and a major public health problem worldwide. Most primary infections with the four dengue virus serotypes (DENV1-4) are inapparent; nonetheless, whether the distribution of symptomatic versus inapparent infections by serotype varies remains unknown. Here, we present (1) the evaluation of a multiplex DENV1-4 envelope domain III multiplex microsphere-based assay (EDIII-MMBA) to serotype inapparent primary infections and (2) its application leveraging 17 years of prospective sample collection from the Nicaraguan Pediatric Dengue Cohort Study (PDCS). First, we evaluated the performance of the EDIII-MMBA with samples characterized by RT-PCR or focus reduction neutralization test. Next, we analyzed 46% (N=574) of total inapparent primary DENV infections in the PDCS with the EDIII-MMBA to evaluate the epidemiology of inapparent infections. Remaining infections were inferred using stochastic imputation, taking year and neighborhood into account. Infection incidence and percentage of inapparent, symptomatic, and severe infections were analyzed by serotype. The EDIII-MMBA demonstrated excellent overall accuracy (100%, 95.8-100%) for serotyping symptomatic and inapparent primary DENV infections when evaluated against gold-standard serotyping methods. We found that a significant majority of primary infections were inapparent, with DENV3 exhibiting the highest likelihood of symptomatic and severe primary infections (Pooled OR compared to DENV1 = 2.13, 95% CI 1.28-3.56, and 6.75, 2.01-22.62, respectively), whereas DENV2 was similar to DENV1 in both analyses. Significant within- and between-year variation in serotype distribution between symptomatic and inapparent infections and circulation of serotypes undetected in symptomatic cases were observed in multiple years. Our study indicates that case surveillance skews the perceived epidemiological footprint of DENV. We reveal a more complex and intricate pattern of serotype distribution in inapparent infections. The significant differences in infection outcomes by serotype emphasizes the need for vaccines with balanced immunogenicity and efficacy across serotypes.

Toothpaste use in Peruvian children during COVID-19 pandemic: results from a National Demographic and Health Survey.

PURPOSE: To determine the impact of COVID-19 pandemic on use of toothpaste in Peruvian children. METHODS: A national database of Peruvian children from 0 to 11 years old was used to develop a cross-sectional study, with a final sample of 51, 013 subjects. Data were obtained of results of the Demographic and Family Health Survey (ENDES); the questionnaire was self-reported. The use of toothpaste and fluoride concentration in toothpaste were dependent variables, and for the independent variable, the year was considered; in addition, other covariates were included. The statistical analyses applied were descriptive, bivariate, and multivariate tests. RESULTS: Use of toothpaste was 98.99% (n = 50,134), while fluoride toothpaste with < 1000 ppm was used by 77.29% (n = 27,366). For bivariate analysis, use of toothpaste was associated with place and area of residence, altitude, natural region, and age; for use of fluoride toothpaste with minimum 1000 ppm, there was an association with place and area of residence, natural region, wealth index, and age. In a multivariate manner, year only presented a positive association with use of fluoride toothpaste < 1000 ppm (RPa:1.04; 95%CI 1.01-1.07). CONCLUSIONS: Year 2020 of COVID-19 pandemic had a positive impact on the use of < 1000 ppm fluoride toothpaste in Peruvian children.

Architecture, Chromatin and Gene Organization of Toxoplasma gondii Subtelomeres.

Subtelomeres (ST) are chromosome regions that separate telomeres from euchromatin and play relevant roles in various biological processes of the cell. While their functions are conserved, ST structure and genetic compositions are unique to each species. This study aims to identify and characterize the subtelomeric regions of the 13 Toxoplasma gondii chromosomes of the Me49 strain. Here, STs were defined at chromosome ends based on poor gene density. The length of STs ranges from 8.1 to 232.4 kbp, with a gene density of 0.049 genes/kbp, lower than the Me49 genome (0.15 kbp). Chromatin organization showed that H3K9me3, H2A.X, and H3.3 are highly enriched near telomeres and the 5' end of silenced genes, decaying in intensity towards euchromatin. H3K4me3 and H2A.Z/H2B.Z are shown to be enriched in the 5' end of the ST genes. Satellite DNA was detected in almost all STs, mainly the sat350 family and a novel satellite named sat240. Beyond the STs, only short dispersed fragments of sat240 and sat350 were found. Within STs, there were 12 functional annotated genes, 59 with unknown functions (Hypothetical proteins), 15 from multigene FamB, and 13 from multigene family FamC. Some genes presented low interstrain synteny associated with the presence of satellite DNA. Orthologues of FamB and FamC were also detected in Neospora caninum and Hammondia hammondi. A re-analysis of previous transcriptomic data indicated that ST gene expression is strongly linked to the adaptation to different situations such as extracellular passage (evolve and resequencing study) and changes in metabolism (lack of acetyl-CoA cofactor). In conclusion, the ST region of the T. gondii chromosomes was defined, the STs genes were determined, and it was possible to associate them with high interstrain plasticity and a role in the adaptability of T. gondii to environmental changes.

Open chromatin analysis in Trypanosoma cruzi life forms highlights critical differences in genomic compartments and developmental regulation at tDNA loci.

BACKGROUND: Genomic organization and gene expression regulation in trypanosomes are remarkable because protein-coding genes are organized into codirectional gene clusters with unrelated functions. Moreover, there is no dedicated promoter for each gene, resulting in polycistronic gene transcription, with posttranscriptional control playing a major role. Nonetheless, these parasites harbor epigenetic modifications at critical regulatory genome features that dynamically change among parasite stages, which are not fully understood. RESULTS: Here, we investigated the impact of chromatin changes in a scenario commanded by posttranscriptional control exploring the parasite Trypanosoma cruzi and its differentiation program using FAIRE-seq approach supported by transmission electron microscopy. We identified differences in T. cruzi genome compartments, putative transcriptional start regions, and virulence factors. In addition, we also detected a developmental chromatin regulation at tRNA loci (tDNA), which could be linked to the intense chromatin remodeling and/or the translation regulatory mechanism required for parasite differentiation. We further integrated the open chromatin profile with public transcriptomic and MNase-seq datasets. Strikingly, a positive correlation was observed between active chromatin and steady-state transcription levels. CONCLUSION: Taken together, our results indicate that chromatin changes reflect the unusual gene expression regulation of trypanosomes and the differences among parasite developmental stages, even in the context of a lack of canonical transcriptional control of protein-coding genes.

Dilataciones quísticas congénitas de las vías biliares (segunda parte). Vías biliares extrahepáticas / Congenital bile duct cystic dilatations (part 2). Extrahepatic bile ducts

RESUMEN Las dilataciones quísticas congénitas de la vía biliar extrahepática son infrecuentes, principalmente en Occidente, y afectan sobre todo a niños pero son poco comunes en adultos. El diagnóstico requiere un alto índice de sospecha y suele llevarse a cabo con estudios colangiográficos de los cuales la CRMN es, en la actualidad, el de mayor utilidad. Es conocido el aumento de malignización de estas dilataciones, por lo que el tratamiento completo de la bolsa quística es obligatorio.

ABSTRACT Congenital dilatation of extrahepatic bile ducts is rare in the Western countries and is more common in children than in adults. The diagnosis requires high level of suspicion and is made by cholangiography tests, among which MRCP is the most useful nowadays. Malignant transformation of these cystic dilatations is well-known; therefore, complete resection of the cystic pouch is mandatory.

Evaluation of Toxoplasma gondii recombinant antigens for early diagnosis of congenital toxoplasmosis.

The performance of Toxoplasma rGra8, rMic1, and the chimeric rGra4-Gra7 antigens for early congenital toxoplasmosis (CT) diagnosis was evaluated. Sera from CT patients showed high IgG reactivity to rMic1, rGra8, and rGra4-Gra7. The seroreactivity of samples from uninfected infants was lost within 2 months of age.

Pensar é uma forma de resistir ao obscurantismo / Thinking is a way of resisting obscurantism / Pensar es una manera de resistirse al oscurantismo / Penser, c'est une manière de résister à l'obscurantisme

Neste texto, traço a relação do tipo de interpretação da obra de arte proposto por J. A. Frayze-Pereira com o que se passa numa sessão de análise. Na sessão, a "obra" do paciente é a narração de seus conteúdos mentais em busca de uma forma simbólica. Nesse processo, sugiro que o que é evocado no analista, que se transforma numa interpretação ou comentário, é central na constituição de seu significado e parte dele. Daí considerarmos a transferência uma poiesis.

In this text I outline the relationship between the type of interpretation of the work of art proposed by the author, and what happens in an psychoanalytic session. In analytic sessions the patient's "work" (oeuvre) is his narrative of his emotional experience in search of a symbolic form. In this process I suggest that what is evoked in the analyst, which becomes an interpretation or comment, is the core to the formation of its meaning and part of it. Hence, we consider the transference as a process of poiesis.

En este texto trazo la relación entre el tipo de interpretación de la obra de arte, propuesta por el autor, con lo que sucede en una sesión de análisis. En la sesión, el "trabajo" del paciente es la narración de sus contenidos mentales en busca de una forma simbólica. En este proceso sugiero que lo evocado en el analista, que se convierte en interpretación o comentario, es central para la constitución de su significado y parte de él. Por tanto, consideramos la transferencia como una poiesis.

Dans ce texte, je trace la relation entre le type d'interprétation de l'Œuvre d'art, proposé par l'auteur, avec ce qui se passe lors d'une séance d'analyse. Dans la séance, le « travail ¼ du patient est la narration de son contenu mental à la recherche d>une forme symbolique. Dans ce processus, je suggère que ce qui est évoqué chez l>analyste, qui devient une interprétation ou un commentaire, est au cŒur de la constitution de son sens et d>une partie de celuici. Nous considérons donc le transfert comme une poïétique - poiesis en grec.

Nucleosome landscape reflects phenotypic differences in Trypanosoma cruzi life forms.

Trypanosoma cruzi alternates between replicative and nonreplicative life forms, accompanied by a shift in global transcription levels and by changes in the nuclear architecture, the chromatin proteome and histone posttranslational modifications. To gain further insights into the epigenetic regulation that accompanies life form changes, we performed genome-wide high-resolution nucleosome mapping using two T. cruzi life forms (epimastigotes and cellular trypomastigotes). By combining a powerful pipeline that allowed us to faithfully compare nucleosome positioning and occupancy, more than 125 thousand nucleosomes were mapped, and approximately 20% of them differed between replicative and nonreplicative forms. The nonreplicative forms have less dynamic nucleosomes, possibly reflecting their lower global transcription levels and DNA replication arrest. However, dynamic nucleosomes are enriched at nonreplicative regulatory transcription initiation regions and at multigenic family members, which are associated with infective-stage and virulence factors. Strikingly, dynamic nucleosome regions are associated with GO terms related to nuclear division, translation, gene regulation and metabolism and, notably, associated with transcripts with different expression levels among life forms. Finally, the nucleosome landscape reflects the steady-state transcription expression: more abundant genes have a more deeply nucleosome-depleted region at putative 5' splice sites, likely associated with trans-splicing efficiency. Taken together, our results indicate that chromatin architecture, defined primarily by nucleosome positioning and occupancy, reflects the phenotypic differences found among T. cruzi life forms despite the lack of a canonical transcriptional control context.

O potencial traumático da pandemia de covid-19 / The trauma potential from covid-19 pandemic / El potencial traumático de la pandemia de covid-19 / Le potentiel traumatique de la pandémie du covid-19

Os autores discutem o potencial traumático desta pandemia, indicando que nem todos os afetados por ela se traumatizarão. A seguir comentam como a experiência da pandemia poderá assumir várias formas simbólicas de acordo com o nível de ansiedade e medo que provocar em cada indivíduo, que será capaz ou não de mobilizar sua capacidade de representação simbólica para integrar em seu psiquismo a experiência dos diversos medos estimulados. São dados exemplos clínicos de algumas dessas expressões simbólicas.

The authors discuss the traumatic potential of this pandemic, showing that not everyone affected by it will be traumatized. After that, they comment on how the experience of the pandemic can take different symbolic forms according to the level of anxiety and fear that it may bring to each individual who will be able or not to mobilize their capacity for symbolic representation to integrate the experience of the different fears brought to their psyche. Three clinical examples of some of these symbolic expressions are given.

Los autores discuten el potencial traumático de esta pandemia, indicando que no todos los afectados por ella quedarán traumatizados. Hacen referencia a cómo la experiencia de la pandemia puede asumir diferentes formas simbólicas según el nivel de ansiedad y miedo que puede causar en cada individuo y que podrá o no movilizar su capacidad de representación simbólica para integrar la experiencia de lo diferente en su psique. Se dan ejemplos clínicos de algunas de esas expresiones simbólicas.

Les auteurs discutent du potentiel traumatique de cette pandémie, ce qui indique que ni tous ceux qui sont atteints ne resteront traumatisés. Ensuite ils commentent comment la pandémie pourra assumer plusieurs formes symboliques selon le niveau d'anxiété et de peur qu'elle provoque chez chaque individu, lequel sera capable, ou non, de mobiliser sa capacité de représentation symbolique pour intégrer dans son psychisme l'expérience des craintes stimulées. On donne des exemples cliniques de quelques-unes de ces expressions symboliques.

Everolimus for cardiac rhabdomyomas in children with tuberous sclerosis. The ORACLE study protocol (everOlimus for caRdiac rhAbdomyomas in tuberous sCLErosis): a randomised, multicentre, placebo-controlled, double-blind phase II trial.

INTRODUCTION: Tuberous sclerosis complex is a rare genetic disorder leading to the growth of hamartomas in multiple organs, including cardiac rhabdomyomas. Children with symptomatic cardiac rhabdomyoma require frequent admissions to intensive care units, have major complications, namely, arrhythmias, cardiac outflow tract obstruction and heart failure, affecting the quality of life and taking on high healthcare cost. Currently, there is no standard pharmacological treatment for this condition, and the management includes a conservative approach and supportive care. Everolimus has shown positive effects on subependymal giant cell astrocytomas, renal angiomyolipoma and refractory seizures associated with tuberous sclerosis complex. However, evidence supporting efficacy in symptomatic cardiac rhabdomyoma is limited to case reports. The ORACLE trial is the first randomised clinical trial assessing the efficacy of everolimus as a specific therapy for symptomatic cardiac rhabdomyoma. METHODS: ORACLE is a phase II, prospective, randomised, placebo-controlled, double-blind, multicentre protocol trial. A total of 40 children with symptomatic cardiac rhabdomyoma secondary to tuberous sclerosis complex will be randomised to receive oral everolimus or placebo for 3 months. The primary outcome is 50% or more reduction in the tumour size related to baseline. As secondary outcomes we include the presence of arrhythmias, pericardial effusion, intracardiac obstruction, adverse events, progression of tumour reduction and effect on heart failure. CONCLUSIONS: ORACLE protocol addresses a relevant unmet need in children with tuberous sclerosis complex and cardiac rhabdomyoma. The results of the trial will potentially support the first evidence-based therapy for this condition.

Toxoplasma gondii seropositivity associated to peri-urban living places in pregnant women in a rural area of Buenos Aires province, Argentina.

Infection with Toxoplasma gondii is very common in humans throughout the world, the intake of raw or undercooked meat with tissue cysts and fruits, vegetables and water contaminated with parasite oocysts being the main routes of infection. Here, we analyzed the seroprevalence of anti-T. gondii antibodies in pregnant females (age 13-44 years; n = 920) between April 2014 and December 2017 from Chascomús (Argentina), a city immersed in a rural area. Altogether 320 tested positive for immunoglobulin G antibodies, yielding an overall seroprevalence of 34.8% (CI 95%: 31.7-37.9). No association was observed between seropositivity and age. In addition, by using the QGIS 3.2.1 software we analyzed the geographical distribution of 769 (83.6%) pregnant females in two main areas of the city: Urban (n = 157) and Peri-urban (n = 612) with a seroprevalence of 26.8% (CI 95%: 19.8-33.7) and 36.4% (CI 95%: 32.6-40.3) respectively, and this difference was statistically significant (p = 0.023). Furthermore, we assessed through a questionnaire survey, between April 2016 to December 2017, possible risk factors such as activity (urban and rural), home water supply, animal husbandry, presence of cats as pets, gardening and consumption of meat and its derivatives (pork, sheep meat and sausages) and their frequencies (consumption per week), not finding significant association with seropositivity. Significant differences was found when the seroprevalence was analyzed between the urban and peri-urban neighborhoods of the city of Chascomús. The higher seroprevalence in peri-urban neighborhoods could be due to an unfavorable socioeconomic situation and/or to undeveloped peri-urban environments, which is a risk factor that should be taken into account when planning the health care of pregnant females.

Association between high-fat diet feeding and male fertility in high reproductive performance mice.

The increasing worldwide prevalence of metabolic syndrome (MetS), especially in younger populations, is a risk factor for fertility disorders. However, a direct correlation of MetS with male infertility still remains unclear. In this work, we evaluated whether MetS has a negative impact on fertility of hybrid male mice with high reproductive performance. To induce a MetS-like condition, (C57BL/6xBALB/c) F1 male mice were fed a high-fat diet (HFD, 30% fat) for 19 weeks, while controls received a normal-fat diet (NFD, 6% fat). HFD-fed animals exhibited increased body weight, hypercholesterolemia, hyperglycemia and glucose intolerance. In vivo fertilisation assays performed along the treatment period showed no differences in fertilisation nor in vitro embryo development rates between groups. While testicular weight and morphology were similar in both groups, HFD-fed mice presented lighter epididymides and higher amounts of gonadal fat. Moreover, sperm count was lower in HFD-fed mice, despite normal sperm viability, morphology, motility or acrosome reaction. Finally, no differences were observed in in vitro fertilisation rates between groups. In summary, although HFD feeding altered some reproductive parameters, it did not impair male fertility in high performance breeders suggesting the possibility that a fertility impairment could be the result of the cumulative combination of environmental and/or genetic factors.

Raman Spectroscopy and Chemometric Modeling to Predict Physical-Chemical Honey Properties from Campeche, Mexico.

In this work, 10 chemometric models based on Raman spectroscopy were constructed to predict the physicochemical properties of honey produced in the state of Campeche, Mexico. The properties of honey studied were pH, moisture, total soluble solids (TSS), free acidity, lactonic acidity, total acidity, electrical conductivity, Redox potential, hydroxymethylfurfural (HMF), and ash content. These proprieties were obtained according to the methods described by the Association of Official Analytical Chemists, Codex Alimentarius, and the International Honey Commission. For the construction of the chemometric models, 189 honey samples were collected and analyzed in triplicate using Raman spectroscopy to generate the matrix data [X], which were correlated with each of the physicochemical properties [Y]. The predictive capacity of each model was determined by cross validation and external validation, using the statistical parameters: standard error of calibration (SEC), standard error of prediction (SEP), coefficient of determination of cross-validation (R2cal), coefficient of determination for external validation (R2val), and Student's t-test. The statistical results indicated that the chemometric models satisfactorily predict the humidity, TSS, free acidity, lactonic acidity, total acidity, and Redox potential. However, the models for electric conductivity and pH presented an acceptable prediction capacity but not adequate to supply the conventional processes, while the models for predicting ash content and HMF were not satisfactory. The developed models represent a low-cost tool to analyze the quality of honey, and contribute significantly to increasing the honey distribution and subsequently the economy of the region.

Experimental evidences that P21 protein controls Trypanosoma cruzi replication and modulates the pathogenesis of infection.

P21 is a protein secreted by Trypanosoma cruzi (T. cruzi). Previous studies have shown a spectrum of biological activities performed by P21 such as induction of phagocytosis, leukocyte chemotaxis and inhibition of angiogenesis. However, the activity of P21 in T. cruzi infection remains unknown. Here, we reported the role of P21 in mice harboring late T. cruzi infection. Treatment with recombinant P21 protein (rP21) reduced parasite load and angiogenesis, and induced fibrosis in the cardiac tissue of infected mice. In addition, rP21 reduced the growth of epimastigotes, inhibited intracellular replication of amastigotes and modulated the parasite cell cycle. Our data suggest that P21 controls parasite replication in the host, supporting the survival of both parasite and host.

Surface-enhanced raman spectroscopy: A non invasive alternative procedure for early detection in childhood asthma biomarkers in saliva.

The early detection of bronchial inflammation in asthma, through a non-invasive, simple method and under a subclinical state, could lead to a more effective control of this condition. The aim of this study was to identify biomarkers of bronchial inflammation in the saliva of children with asthma through immunoassay and Surface Enhanced Raman Spectroscopy (SERS). We conducted an analytical cross-sectional study in 44 children ages 6-12; the diagnosis of asthma was made according to Global Initiative for Asthma (GINA) standards. The children's saliva was analyzed by immunoassay for the quantification of 37 cytokines, as well as SERS analysis in a confocal Raman microscope at 785 nm. We found a significant association between bronchial obstruction and IL-8 (p = 0.004), IL-10 (p = 0.008) and sCD163 (p = 0.003). The Raman spectra showed significant amplification in the region of 760 to 1750 cm-1. The Principal Component Analysis and Linear Discriminant Analysis (PCA-LDA) method has a sensitivity of 85%, specificity of 82% and an accuracy of 84% for the diagnosis of asthma. These results demonstrate the presence of a subclinical inflammatory state, suggestive of bronchial remodeling in the population studied. The SERS method is a potential tool for identifying bronchial inflammation and its endotype, allowing for a highly sensitive and specific diagnosis.

Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruzi.

DNA polymerase theta (Polθ), a member of the DNA polymerase family A, exhibits a polymerase C-terminal domain, a central domain, and an N-terminal helicase domain. Polθ plays important roles in DNA repair via its polymerase domain, regulating genome integrity. In addition, in mammals, Polθ modulates origin firing timing and MCM helicase recruitment to chromatin. In contrast, as a model eukaryote, Trypanosoma cruzi exhibits two individual putative orthologs of Polθ in different genomic loci; one ortholog is homologous to the Polθ C-terminal polymerase domain, and the other is homologous to the Polθ helicase domain, called Polθ-polymerase and Polθ-helicase, respectively. A pull-down assay using the T. cruzi component of the prereplication complex Orc1/Cdc6 as bait captured Polθ-helicase from the nuclear extract. Orc1/Cdc6 and Polθ-helicase directly interacted, and Polθ-helicase presented DNA unwinding and ATPase activities. A T. cruzi strain overexpressing the Polθ-helicase domain exhibited a significantly decreased amount of DNA-bound MCM7 and impaired replication origin firing. Taken together, these data suggest that Polθ-helicase modulates DNA replication by directly interacting with Orc1/Cdc6, which reduces the binding of MCM7 to DNA and thereby impairs the firing of replication origins.

High species richness of native pollinators in Brazilian tomato crops / Alta riqueza de espécies de polinizadores nativos em plantações Brasileiras de tomateiro

Abstract Pollinators provide an essential service to natural ecosystems and agriculture. In tomatoes flowers, anthers are poricidal, pollen may drop from their pore when flowers are shaken by the wind. However, bees that vibrate these anthers increase pollen load on the stigma and in fruit production. The present study aimed to identify the pollinator richness of tomato flowers and investigate their morphological and functional traits related to the plant-pollinator interaction in plantations of Central Brazil. The time of anthesis, flower duration, and the number and viability of pollen grains and ovules were recorded. Floral visitors were observed and collected. Flower buds opened around 6h30 and closed around 18h00. They reopened on the following day at the same time in the morning, lasting on average 48 hours. The highest pollen availability occurred during the first hours of anthesis. Afterwards, the number of pollen grains declined, especially between 10h00 to 12h00, which is consistent with the pollinator visitation pattern. Forty bee species were found in the tomato fields, 30 of which were considered pollinators. We found that during the flowering period, plants offered an enormous amount of pollen to their visitors. These may explain the high richness and amount of bees that visit the tomato flowers in the study areas. The period of pollen availability and depletion throughout the day overlapped with the bees foraging period, suggesting that bees are highly effective in removing pollen grains from anthers. Many of these grains probably land on the stigma of the same flower, leading to self-pollination and subsequent fruit development. Native bees (Exomalopsis spp.) are effective pollinators of tomato flowers and are likely to contribute to increasing crop productivity. On the other hand, here tomato flowers offer large amounts of pollen resource to a high richness and amount of bees, showing a strong plant-pollinator interaction in the study agroecosystem.

Resumo Polinizadores fornecer um serviço essencial para os ecossistemas naturais e para agricultura. Em tomateiros, as anteras são poricidas e o pólen pode sair a partir dos poros quando as flores são agitadas pelo vento. No entanto, as abelhas que vibram as anteras aumentam a carga de pólen no estigma e na produção de frutos. O presente estudo teve como objetivo identificar a riqueza dos polinizadores das flores de tomate e investigar suas características morfológicas e funcionais relacionadas com a interação planta-polinizador em plantações do Brasil central. Foram registrados o tempo de antese, duração flor, bem como o número e viabilidade de grãos de pólen e óvulos. Os visitantes florais foram observados e coletados. Os botões florais abriram-se em torno 06h30 e fechou em torno de 18h00. As flores reabrem no dia seguinte ao mesmo tempo na parte da manhã, com longevidade média de 48 horas. A maior disponibilidade de pólen ocorreu durante as primeiras horas da antese. Depois disso, o número de grãos de pólen diminuiu, especialmente entre as 10h00 às 12h00, o que é consistente com os padrões de visitação de polinizadores. Quarenta espécies de abelhas foram encontradas nos campos de tomate, 30 das quais foram consideradas polinizadores. Durante o período de floração, as plantas oferecem enorme quantidade de pólen para os seus visitantes. Isto pode explicar a alta riqueza e quantidade de abelhas que visitam as flores de tomate nas áreas de estudo. O período de disponibilidade e redução de pólen durante todo o período do dia é sobreposto com o período de alimentação das abelhas, o que sugere que elas são altamente eficazes na remoção de grãos de pólen das anteras. Muitos desses grãos provavelmente são depositados no estigma da mesma flor, levando à auto-polinização e o desenvolvimento de frutos. Abelhas nativas (Exomalopsis spp.) são polinizadores efetivos de flores de tomate, podendo contribuir para o aumento da produtividade das culturas. Por outro lado, as flores de tomate oferecem grandes quantidades de pólen de recursos para uma alta riqueza e quantidade de abelhas, que mostram um estudo forte interação planta-polinizador nos agroecossistemas.

Single molecule force spectroscopy reveals the effect of BiP chaperone on protein folding.

BiP (Immunoglobulin Binding Protein) is a member of the Hsp70 chaperones that participates in protein folding in the endoplasmic reticulum. The function of BiP relies on cycles of ATP hydrolysis driving the binding and release of its substrate proteins. It still remains unknown how BiP affects the protein folding pathway and there has been no direct demonstration showing which folding state of the substrate protein is bound by BiP, as previous work has used only peptides. Here, we employ optical tweezers for single molecule force spectroscopy experiments to investigate how BiP affects the folding mechanism of a complete protein and how this effect depends on nucleotides. Using the protein MJ0366 as the substrate for BiP, we performed pulling and relaxing cycles at constant velocity to unfold and refold the substrate. In the absence of BiP, MJ0366 unfolded and refolded in every cycle. However, when BiP was added, the frequency of folding events of MJ0366 significantly decreased, and the loss of folding always occurred after a successful unfolding event. This process was dependent on ATP and ADP, since when either ATP was decreased or ADP was added, the duration of periods without folding events increased. Our results show that the affinity of BiP for the substrate protein increased in these conditions, which correlates with previous studies in bulk. Therefore, we conclude that BiP binds to the unfolded state of MJ0366 and prevents its refolding, and that this effect is dependent on both the type and concentration of nucleotides.