Landscape dynamics and diversification of the megadiverse South American freshwater fish fauna.

Landscape dynamics are widely thought to govern the tempo and mode of continental radiations, yet the effects of river network rearrangements on dispersal and lineage diversification remain poorly understood. We integrated an unprecedented occurrence dataset of 4,967 species with a newly compiled, time-calibrated phylogeny of South American freshwater fishes-the most species-rich continental vertebrate fauna on Earth-to track the evolutionary processes associated with hydrogeographic events over 100 Ma. Net lineage diversification was heterogeneous through time, across space, and among clades. Five abrupt shifts in net diversification rates occurred during the Paleogene and Miocene (between 30 and 7 Ma) in association with major landscape evolution events. Net diversification accelerated from the Miocene to the Recent (c. 20 to 0 Ma), with Western Amazonia having the highest rates of in situ diversification, which led to it being an important source of species dispersing to other regions. All regional biotic interchanges were associated with documented hydrogeographic events and the formation of biogeographic corridors, including the Early Miocene (c. 23 to 16 Ma) uplift of the Serra do Mar and Serra da Mantiqueira and the Late Miocene (c. 10 Ma) uplift of the Northern Andes and associated formation of the modern transcontinental Amazon River. The combination of high diversification rates and extensive biotic interchange associated with Western Amazonia yielded its extraordinary contemporary richness and phylogenetic endemism. Our results support the hypothesis that landscape dynamics, which shaped the history of drainage basin connections, strongly affected the assembly and diversification of basin-wide fish faunas.

Mutations in trpγ, the homologue of TRPC6 autism candidate gene, causes autism-like behavioral deficits in Drosophila.

Autism Spectrum Disorder (ASD) is characterized by impaired social communication, restricted interests, and repetitive and stereotyped behaviors. The TRPC6 (transient receptor potential channel 6) represents an ASD candidate gene under an oligogenic/multifactorial model based on the initial description and cellular characterization of an individual with ASD bearing a de novo heterozygous mutation disrupting TRPC6, together with the enrichment of disruptive TRPC6 variants in ASD cases as compared to controls. Here, we perform a clinical re-evaluation of the initial non-verbal patient, and also present eight newly reported individuals ascertained for ASD and bearing predicted loss-of-function mutations in TRPC6. In order to understand the consequences of mutations in TRPC6 on nervous system function, we used the fruit fly, Drosophila melanogaster, to show that null mutations in transient receptor gamma (trpγ; the fly gene most similar to TRPC6), cause a number of behavioral defects that mirror features seen in ASD patients, including deficits in social interactions (based on courtship behavior), impaired sleep homeostasis (without affecting the circadian control of sleep), hyperactivity in both young and old flies, and defects in learning and memory. Some defects, most notably in sleep, differed in severity between males and females and became normal with age. Interestingly, hyperforin, a TRPC6 agonist and the primary active component of the St. John's wort antidepressant, attenuated many of the deficits expressed by trpγ mutant flies. In summary, our results provide further evidence that the TRPC6 gene is a risk factor for ASD. In addition, they show that the behavioral defects caused by mutations in TRPC6 can be modeled in Drosophila, thereby establishing a paradigm to examine the impact of mutations in other candidate genes.

Guidance provided by pharmacists to customers regarding to destination of unused household medications: disposal of household medications.

BACKGROUND: Discarding pharmaceuticals in the garbage or into the sewage system are still the most common methods in many countries. This study aims to investigate the guidance provided by pharmacists to customers on the disposal of unused and expired household medications in São Paulo State, Brazil. METHOD: The study population consisted of 630 pharmacists from the State of São Paulo, who work in community pharmacies. They answered an online questionnaire with questions composed in three blocks: demographic, work, and academic information on the pharmacist; guidance about the disposal of household medications; and knowledge regarding the reverse logistics of these medications. An invitation to participate in the questionnaire was made via WhatsApp, individually and collectively. Inferential statistics were performed using the chi-square test and were considered significant when p < 0.05%. RESULTS: Among the participating pharmacists, the majority were women under 60 years old,56 (8.89%) stated that they never orient the customer regarding the disposal of unused and expired household medications, while 574 (91,12%) indicated that they almost provide guidance. The frequency with which they provided guidance was influenced by the number of years since graduation (p = 0.0047), the time they had worked in pharmacies and drugstores (p = 0.0007), and whether or not they had a graduate degree (p = 0.0181). Regarding the disposal of medications, among the 643 responses provided by the pharmacists,516 (80.25%) indicated that they oriented customers to return them to a pharmacy. CONCLUSION: A small number of pharmacists always orient customers on the proper disposal that should be followed for unused and expired household medications, prioritizing their return to a pharmacy. In general, these pharmacists have longer periods of work experience and higher academic qualifications. Thus, it is important to increase knowledge through professional training and further education programs.

Efficacy of Carotenoid-Loaded Gelatin Nanoparticles in Reducing Plasma Cytokines and Adipocyte Hypertrophy in Wistar Rats.

The present study investigated the effect of gelatin-based nanoparticles (EPG) loaded with a carotenoid-rich crude extract (CE) on systemic and adipose tissue inflammatory response in a model with inflammation induced by a high glycemic index and high glycemic load diet (HGLI). Nanoparticles synthesized were characterized by different physical and chemical methods. The in vivo investigation evaluated Wistar rats (n = 20, 11 days, adult male with 21 weeks) subdivided into untreated (HGLI diet), conventional treatment (nutritionally adequate diet), treatment 1 (HGLI + crude extract (12.5 mg/kg)), and treatment 2 (HGLI + EPG (50 mg/kg)) groups. Dietary intake, caloric intake and efficiency, weight, inflammatory cytokines tissue concentration, visceral adipose tissue (VAT) weight, histopathological analysis, and antioxidant activity in plasma and VAT were investigated. EPG showed the same physical and chemical characteristics as previous batches (95.2 nm, smooth surface, and chemical interactions between materials). The EPG-treated group was the only group promoting negative ∆dietary intake, ∆caloric efficiency, and ∆weight. In addition, it presented a significant reduction (p < 0.05) in IL-6 and leptin levels and a greater presence of multilocular adipocytes. The results suggest that EPG can act as a nutraceutical in adjuvant therapy for treating inflammatory diseases associated with adipose tissue accumulation.

Inhibition of receptor-binding domain-ACE2 interaction after two doses of Sinovac's CoronaVac or AstraZeneca/Oxford's AZD1222 SARS-CoV-2 vaccines.

Practical laboratory proxies that correlate to vaccine efficacy may facilitate trials, identify nonresponders, and inform about boosting strategies. Among clinical and laboratory markers, assays that evaluate antibodies that inhibit receptor-binding domain (RBD) ligation to angiotensin-converting enzyme-2 receptor (receptor-binding inhibition [RBI]) may provide a surrogate for viral neutralization assays. We evaluated RBI before and after a median of 34 days (interquartile range [IQR]: 33-40) of the second dose of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Sinovac's CoronaVac (CN) or AstraZeneca/Oxford's AZD1222 (AZ) vaccines in 166 individuals. Both vaccines elicited high inhibitory titers in most subjects, 95% (158/166), with signal inhibition above 30% and 89% (127/143) with more than fourfold increase from prevaccination titers, but titers tend to decrease over time. Both postvaccination inhibitory titers (95%, IQR 85%-97% for AZ vs. 79%, IQR 60%-96% for CN, p = 0.004) and pre/post-titer increase (AZ 76%, IQR 51%-86% for AZ vs. 47%, IQR 24%-67% for CN, p < 0.0001) were higher among AZ vaccinees. Previous serological reactivity due to natural infection was associated with high prevaccination signal inhibition titers. The study documents a robust antibody response capable of interfering with RBD-angiotensin-converting enzyme binding. Evaluation of SARS-CoV-2 infection incidence in these populations is necessary to assess its association to protection and its duration.

A simple algorithm for selecting cases to investigate acute and early HIV infections in low- and middle-income countries.

We documented the outcome of an over 10-year (2011-2021) effort to diagnose acute and early HIV infections (AEHI) in an Infectious Diseases Outpatient Clinic with limited resources. Of a total of 132, 119 HIV-RNA tests were performed from 2017 to 2020, 12 cases were identified, using a simple algorithm: risk exposure of 6 weeks or less before the visit and/or symptoms compatible with acute retroviral syndrome 7-30 days after exposure and/or undetermined 3rd generation rapid diagnostic test or serology. AEHI diagnoses varied from 2.4% among asymptomatic to 25% for undetermined serology cases using this simple screening applicable to different settings.

Humoral response to spike S1 and S2 and nucleocapsid proteins on microarray after SARS-CoV-2 infection.

Many aspects of the humoral immune response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), such as its role in protection after natural infection, are still unclear. We evaluated IgA and IgG response to spike subunits 1 and 2 (S1 and S2) and Nucleocapsid proteins of SARS-COV-2 in serum samples of 109 volunteers with viral RNA detected or seroconversion with different clinical evolution (asymptomatic, mild, moderate, and severe coronavirus disease 2019), using the ViraChip® Test Kit. We observed that the quantification of antibodies to all antigens had a positive correlation to disease severity, which was strongly associated with the presence of comorbidities. Seroreversion was not uncommon even during the short (median of 77 days) observation, occurring in 15% of mild-asymptomatic cases at a median of 55 days for IgG and 46 days for IgA. The time to reach the maximal antibody response did not differ significantly among recovered and deceased volunteers. Our study illustrated the dynamic of anti-S1, anti-N, and anti-S2 IgA and IgG antibodies, and suggests that high production of IgG and IgA does not guarantee protection to disease severity and that functional responses that have been studied by other groups, such as antibody avidity, need further attention.

The economic burden of malaria: a systematic review.

BACKGROUND: Quantifying disease costs is critical for policymakers to set priorities, allocate resources, select control and prevention strategies, and evaluate the cost-effectiveness of interventions. Although malaria carries a very large disease burden, the availability of comprehensive and comparable estimates of malaria costs across endemic countries is scarce. METHODS: A literature review to summarize methodologies utilized to estimate malaria treatment costs was conducted to identify gaps in knowledge. RESULTS: Only 45 publications met the inclusion criteria. They utilize different methods, include distinct cost components, have varied geographical coverage (a country vs a city), include different periods in the analysis, and focus on specific parasite types or population groups (e.g., pregnant women). CONCLUSIONS: Cost estimates currently available are not comparable, hindering broad statements on the costs of malaria, and constraining advocacy efforts towards investment in malaria control and elimination, particularly with the finance and development sectors of the government.

Impact of pelvic floor muscle strength on female sexual function: retrospective multicentric cross-sectional study.

INTRODUCTION AND HYPOTHESIS: The female pelvic floor muscles (PFM) play an important role in sexual function. We hypothesize that there is a relation between PFM strength and sexual function in women with and without sexual dysfunction. METHODS: Retrospective multicentric cross-sectional study including women with and without sexual dysfunction [Female Sexual Function Index (FSFI) cutoff point < 26.55], analyzed by age range, considering demographic, clinical, anthropometric and PFM strength [Modified Oxford Scale (MOS)] data. Chi-square, Mann-Whitney test and linear regression (ranks) were used, with 5% significance (SAS 9.4). RESULTS: Out of 1013 medical records, 982 women with an average age of 45.76 (± 15.25) were included in the study. Of these, 679 (69.14%) presented FSFI score < 26.55, while 303 (30.86%) presented FSFI ≥ 26.55. It was identified that the higher scores were among white women, < 45 years old, single, with higher education, family income > 4 minimum wages, body mass index < 25 kg/m2, lower parity, regular physical activity practitioner and higher PFM strength (MOS: 4-5). Desire, arousal, lubrication and orgasm domains were higher among women with MOS 4-5, while satisfaction and pain domains were higher among those with MOS 3-5. CONCLUSION: Demographic, clinical and anthropometric conditions can influence both PFM strength and female sexual function. Our findings demonstrate that women with higher PFM strength present fewer complaints about sexual dysfunction.

Testing, Inequities and Vulnerability of Adolescents to Sexually Transmitted Infections.

Sexually transmitted infections (STIs) should form part of the comprehensive healthcare agenda for adolescents and young people (A&Y) and testing plays a strategic role in STI prevention. However, little attention has been paid to the social and programmatic obstacles to this practice among A&Y. The aim of this study is to describe the prevalence of testing for syphilis and HIV reported by a group of A&Y, analyzing its relationship with variables potentially indicative of inequities related to vulnerability to STIs. This was a cross-sectional survey applied to students at public high schools in three cities in the state of São Paulo, Brazil. The responses of 369 young people who reported having started sexual life were analyzed. The analysis included description of the frequencies of demographic and socioeconomic variables and their associations with testing for syphilis and HIV through the chi-square test. The concentration index (CI) was also calculated, with construction of the respective concentration curve for the associated variables. Positive associations were found between occurrence of testing and living in homes with up to three inhabitants, living in a marital situation with a regular or committed partnership, previous use of primary care services and previous use of hospitals or emergency services. The concentration curve for testing STIs showed significant contributions from the associated variables. The findings indicated that situations of inequity need to be addressed in order to reduce A&Y's vulnerability to STIs.

Interaction between the impact of the Coronavirus disease 2019 pandemic and demographic characteristics on sexual/erectile dysfunction in Latin America: cross-sectional study.

AIM: Our objective was to investigate whether there is an interaction between the COVID-19 pandemic, demographic characteristics and erectile/sexual (E/S) function in individuals from Latin America. MATERIALS AND METHODS: Cross-sectional study which included Latin American individuals over 18 years old, recruited through social media and interviewed between July and August 2020 by online surveys (Google Forms) in Portuguese and Spanish languages. The E/S function was evaluated through the following questionnaires: Simplified International Index of Erectile Function (IIEF-5) and Female Sexual Function Index (FSFI); while post-traumatic stress disorder (PTSD) triggered by the COVID-19 pandemic was assessed through the Impact of Event Scale Revised (IES-R). The data was analyzed by T Student, bivariate and multivariate logistic regression, with significance determined by the Wald test (p<0.05), using the R software v4.0. RESULTS: Out of the 2016 individuals that responded to the survey, 1986 were included and 743 of them presented E/S dysfunction. PTSD occurrence was greater among people with E/S dysfunction when compared to those without E/S dysfunction, in the total score (males: IES-R=26.54[±19.17] and females: IES-R=35.92[±19.25]) and also in the three domains. It was found that those who do not live with a partner were 74% more likely to have E/S dysfunction, but living with a partner during the pandemic had a greater impact on E/S function. CONCLUSION: A negative interaction between the impact of the COVID-19 pandemic and erectile/sexual function of the Latin American population was observed, with greater implications among the individuals who live with their partners.

The impact of the gut microbiome on memory and sleep in Drosophila.

The gut microbiome has been proposed to influence diverse behavioral traits of animals, although the experimental evidence is limited and often contradictory. Here, we made use of the tractability of Drosophila melanogaster for both behavioral analyses and microbiome studies to test how elimination of microorganisms affects a number of behavioral traits. Relative to conventional flies (i.e. with unaltered microbiome), microbiologically sterile (axenic) flies displayed a moderate reduction in memory performance in olfactory appetitive conditioning and courtship assays. The microbiological status of the flies had a small or no effect on anxiety-like behavior (centrophobism) or circadian rhythmicity of locomotor activity, but axenic flies tended to sleep for longer and displayed reduced sleep rebound after sleep deprivation. These last two effects were robust for most tests conducted on both wild-type Canton S and w1118 strains, as well for tests using an isogenized panel of flies with mutations in the period gene, which causes altered circadian rhythmicity. Interestingly, the effect of absence of microbiota on a few behavioral features, most notably instantaneous locomotor activity speed, varied among wild-type strains. Taken together, our findings demonstrate that the microbiome can have subtle but significant effects on specific aspects of Drosophila behavior, some of which are dependent on genetic background.

Vigna spp. Root-Nodules Harbor Potentially Pathogenic Fungi Controlled By Co-habiting Bacteria.

This study aimed to isolate, identify, and evaluate the pathogenicity of nodule-borne fungi of asymptomatic Vigna spp. plants, grown in soils from preserved tropical dry forests (Caatinga) areas and identify the occurrence of co-habiting bacteria from these plants, and which have potential to control the co-occurring pathogenic fungi. Fungi and bacteria were isolated from three Vigna species (V. unguiculata, V. radiata, and V. mungo), grown in soil samples collected in five preserved Caatinga areas (Northeastern, Brazil). All fungi and selected bacteria were phylogenetically characterized by the sequencing of ITS1-5.8S-ITS2, and the 16S rRNA gene, respectively. The pathogenicity of fungi in cowpea seeds germination was evaluated throughout the inoculation experiment in Petri dishes and pots containing sterile substrate. The potential of nodule-borne bacteria to control pathogenic fungi in cowpea was assessed in a pot experiment with a sterilized substrate by the co-inoculation of fungi and bacteria isolated from the respective individual plants and soils. The 23 fungal isolates recovered were classified within the genera Fusarium, Macrophomina, Aspergillus, Cladosporium, and Nigrospora. The inoculation of fungi in cowpea seeds reduced the emergence of seeds in Petri dishes and pots. Twenty-four bacteria (Agrobacterium sp., Bradyrhizobium sp., Bacillus sp., Enterobacter sp., Pseudomonas sp., Paraburkholderia sp., and Rhizobium sp.) inhibited the harmful effects of Macrophomina sp. and Fusarium sp., increasing the germination and emergency of potted cowpea plants, highlighting the strains Agrobacterium sp. ESA 686 and Pseudomonas sp. ESA 732 that controlled, respectively, the Fusarium sp. ESA 771 and Macrophomina sp. ESA 786 by 100 and 84.6% of efficiency.

In Silico Study, Physicochemical, and In Vitro Lipase Inhibitory Activity of α,ß-Amyrenone Inclusion Complexes with Cyclodextrins.

α,ß-amyrenone (ABAME) is a triterpene derivative with many biological activities; however, its potential pharmacological use is hindered by its low solubility in water. In this context, the present work aimed to develop inclusion complexes (ICs) of ABAME with γ- and ß-cyclodextrins (CD), which were systematically characterized through molecular modeling studies as well as FTIR, XRD, DSC, TGA, and SEM analyses. In vitro analyses of lipase activity were performed to evaluate possible anti-obesity properties. Molecular modeling studies indicated that the CD:ABAME ICs prepared at a 2:1 molar ratio would be more stable to the complexation process than those prepared at a 1:1 molar ratio. The physicochemical characterization showed strong evidence that corroborates with the in silico results, and the formation of ICs with CD was capable of inducing changes in ABAME physicochemical properties. ICs was shown to be a stronger inhibitor of lipase activity than Orlistat and to potentiate the inhibitory effects of ABAME on porcine pancreatic enzymes. In conclusion, a new pharmaceutical preparation with potentially improved physicochemical characteristics and inhibitory activity toward lipases was developed in this study, which could prove to be a promising ingredient for future formulations.

Increase in the Antioxidant and Anti-Inflammatory Activity of Euterpe oleracea Martius Oil Complexed in ß-Cyclodextrin and Hydroxypropyl-ß-Cyclodextrin.

Reactive oxygen species (ROS) are aerobic products generated during cellular respiration, but in the case of oxidative stress, they become key factors in the development of inflammatory processes and chronic diseases such as diabetes and rheumatoid arthritis. In this work, Euterpe oleracea oil (EOO), as well as the complexes produced by slurry (S) and kneading (K), were analyzed for antioxidant capacity in vitro, while only the ß-cyclodextrin complex obtained by kneading (EOO-ßCD-K), which showed better complexation, was selected for anti-inflammatory assays in vivo. In the scavenging activity of OH·, the hydroxypropyl-ß-cyclodextrin complex obtained by kneading (EOO-HPßCD-K) exhibited an activity 437% higher than the pure oil. In the paw edema assay, EOO-ßCD-K reduced edema by 200% and myeloperoxidase (MPO) activity by 112%. In an air pouch model, this treatment showed a reduction in leukocyte, MPO, and Interleukin-1ß (IL-1ß) levels; meanwhile those of glutathione and IL-10 were increased, demonstrating its ability to potentiate the anti-inflammatory effect of EOO.

Meta-analysis of the differential effects of habitat fragmentation and degradation on plant genetic diversity.

Genetic diversity is a key factor for population survival and evolution. However, anthropogenic habitat disturbance can erode it, making populations more prone to extinction. Aiming to assess the global effects of habitat disturbance on plant genetic variation, we conducted a meta-analysis based on 92 case studies obtained from published literature. We compared the effects of habitat fragmentation and degradation on plant allelic richness and gene diversity (equivalent to expected heterozygosity) and tested whether such changes are sensitive to different life-forms, life spans, mating systems, and commonness. Anthropogenic disturbance had a negative effect on allelic richness, but not on gene diversity. Habitat fragmentation had a negative effect on genetic variation, whereas habitat degradation had no effect. When we examined the individual effects in fragmented habitats, allelic richness and gene diversity decreased, but this decrease was strongly dependent on certain plant traits. Specifically, common long-lived trees and self-incompatible species were more susceptible to allelic richness loss. Conversely, gene diversity decreased in common short-lived species (herbs) with self-compatible reproduction. In a wider geographical context, tropical plant communities were more sensitive to allelic richness loss, whereas temperate plant communities were more sensitive to gene diversity loss. Our synthesis showed complex responses to habitat disturbance among plant species. In many cases, the absence of effects could be the result of the time elapsed since the disturbance event or reproductive systems favoring self-pollination, but attention must be paid to those plant species that are more susceptible to losing genetic diversity, and appropriate conservation should be actions taken.

Meta-Análisis de los Efectos Diferenciales de la Fragmentación y Degradación del Hábitat sobre la Diversidad Genética de las Plantas Resumen La diversidad genética es un factor clave para la supervivencia y evolución de las poblaciones. Sin embargo, la perturbación antropogénica de los hábitats puede dañar esta diversidad, volviendo a las poblaciones más susceptibles a la extinción. Con el objetivo de evaluar los efectos globales de la perturbación del hábitat sobre la variación genética de las plantas, realizamos un meta-análisis basado en 92 estudios de caso obtenidos de la literatura publicada. Comparamos los efectos de la degradación y fragmentación del hábitat sobre la riqueza de alelos y la diversidad de genes (equivalente a la heterocigosidad esperada) de las plantas y probamos si dichos cambios son sensibles para diferentes formas de vida, tiempos de vida, sistemas de apareamiento y preponderancia. La perturbación antropogénica tuvo un efecto negativo sobre la riqueza de alelos, pero no sobre la diversidad genética. La fragmentación del hábitat tuvo un efecto negativo sobre la variación genética, mientras que la degradación del hábitat no tuvo efecto. Cuando examinamos los efectos individuales en los hábitats fragmentados, la riqueza de alelos y la diversidad genética disminuyeron, pero esta disminución estuvo vinculada fuertemente con ciertas características de las plantas. Específicamente, los árboles de larga vida y las especies auto-incompatibles fueron más susceptibles a la pérdida de la riqueza de alelos. De manera contraria, la diversidad genética disminuyó en especies comunes de vida corta (hierbas) con reproducción auto-compatibles. En un contexto geográfico más amplio, las comunidades de plantas tropicales fueron más sensibles a la pérdida de la riqueza de alelos, mientras que las comunidades de plantas de zonas templadas fueron más sensibles a la pérdida de diversidad de genes. Nuestra síntesis mostró respuestas complejas a la perturbación del hábitat entre las especies de plantas. En muchos casos, la ausencia de efectos podría ser el resultado del tiempo transcurrido desde el evento de perturbación o los sistemas reproductivos que favorecen la auto-polinización, pero se le debe prestar atención a aquellas especies de plantas que son más susceptibles a la pérdida de la diversidad genética, para así poder realizar las acciones de conservación apropiadas.

Toxoplasma gondii infection damages the perineuronal nets in a murine model.

BACKGROUND: Behavioral and neurochemical alterations associated with toxoplasmosis may be influenced by the persistence of tissue cysts and activation of an immune response in the brain of Toxoplasma gondii-infected hosts. The cerebral extracellular matrix is organised as perineuronal nets (PNNs) that are both released and ensheath by some neurons and glial cells. There is evidences to suggest that PNNs impairment is a pathophysiological mechanism associated with neuropsychiatric conditions. However, there is a lack of information regarding the impact of parasitic infections on the PNNs integrity and how this could affect the host's behavior. OBJECTIVES: In this context, we aimed to analyse the impact of T. gondii infection on cyst burden, PNNs integrity, and possible effects in the locomotor activity of chronically infected mice. METHODS: We infected mice with T. gondii ME-49 strain. After thirty days, we assessed locomotor performance of animals using the open field test, followed by evaluation of cysts burden and PNNs integrity in four brain regions (primary and secondary motor cortices, prefrontal and somesthetic cortex) to assess the PNNs integrity using Wisteria floribunda agglutinin (WFA) labeling by immunohistochemical analyses. FINDINGS AND MAIN CONCLUSIONS: Our findings revealed a random distribution of cysts in the brain, the disruption of PNNs surrounding neurons in four areas of the cerebral cortex and hyperlocomotor behavior in T. gondii-infected mice. These results can contribute to elucidate the link toxoplasmosis with the establishment of neuroinflammatory response in neuropsychiatric disorders and to raise a discussion about the mechanisms related to changes in brain connectivity, with possible behavioral repercussions during chronic T. gondii infection.

Extracellular vesicles isolated from Toxoplasma gondii induce host immune response.

This study established a protocol to purify Toxoplasma gondii tachyzoite microvesicles and exosomes, called as extracellular vesicles (EVs). In addition, the investigations were conducted to determine the kinetic of EV release by tachyzoites and whether EV proteins are able to modulate the host immune response. The particle size and concentration released by tachyzoites in culture medium at different incubation-period were characterized by nanoparticle tracking analysis. Tachyzoites (1 × 106 ) released around 4.37 ± 0.81 × 108 EVs/mL/h, with size varying between 138.2 and 171.9 nm. EVs released into the medium were purified by gel-exclusion chromatography and screened by ELISA, using a pool of human positive sera for toxoplasmosis. EV-fractions contained high concentration of proteins, and EVs were analyzed by scanning and transmission electron microscopies. Tachyzoites released EVs into the culture medium throughout all membrane surface, and these vesicles contain small RNAs/miRNA. Pooled sera from chronically infected human or mice (infected with 2 different T. gondii strains) recognized distinct EV electrophoretic patterns in immunoblotting. T. gondii EVs significantly induced IL-10, TNF-α and iNOS in murine macrophages. In conclusion, this study shows that T. gondii secrete/excrete EVs (microvesicles and exosomes) contain miRNA and they were immunologically recognized by host immune response.

Gastroprotective and Antioxidant Activity of Kalanchoe brasiliensis and Kalanchoe pinnata Leaf Juices against Indomethacin and Ethanol-Induced Gastric Lesions in Rats.

Kalanchoe brasiliensis and Kalanchoe pinnata are used interchangeably in traditional medicine for treating peptic ulcers and inflammatory problems. In this context, this study aims to characterize the chemical constituents and evaluate the gastroprotective activity of the leaf juices of the two species in acute gastric lesions models. Thin Layer Chromatography (TLC) and Ultra High Performance Liquid Chromatography coupled to Mass Spectrometer (UHPLC-MS) were performed for chemical characterization. Wistar rats were pre-treated orally with leaf juices (125, 250 and 500 mg/kg) or ranitidine (50 mg/kg). The peaks observed in the chromatogram of K. brasiliensis showed similar mass spectra to flavonoid glycosides derived from patuletin and eupafolin, while K. pinnata showed mass spectra similar to compounds derived from quercetin, patuletin, eupafolin and kaempferol. K. brasiliensis at all doses and K. pinnata at doses of 250 mg/kg and 500 mg/kg significantly reduced the lesions in the ethanol induction model. In the indomethacin induction model, both species showed significant results at doses of 250 and 500 mg/kg. Also, the pre-treatment with leaf juices increased the antioxidant defense system, glutathione (GSH), whereas malondialdehyde (MDA), myeloperoxidase (MPO), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) levels were significantly decreased. Treatment with leaf juices led to the upregulation of zone occludes-1 (ZO-1) and the downregulation of inducible nitric oxide synthase (iNOS) and factor nuclear-κβ transcription (NF-κB-p65), while also showing a cytoprotective effect and maintaining mucus production. These findings show that the leaf juices of the two species showed gastroprotective effects on ethanol and gastric indomethacin injury which were a consequence of gastric inflammation suppression, antioxidant activity and the maintenance of cytoprotective defenses and mucosal structure architecture.

Intestinal Anti-Inflammatory Activity of the Aqueous Extract from Ipomoea asarifolia in DNBS-Induced Colitis in Rats.

Inflammatory bowel disease is triggered by an uncontrolled immune response associated with genetic, environmental, and intestinal microbiota imbalance. Ipomoea asarifolia (IA), popularly known as "salsa" or "brave salsa", belongs to the Convolvulaceae family. The aim of this approach was to study the preventive effect of IA aqueous extract in 2,4-dinitrobenzene sulfonic acid (DNBS)-induced colitis in rats. Rats pretreated with IA extract or sulfasalazine (SSZ) received intracolonic instillation of DNBS in 50% ethanol (v/v). IA extract presented a protective effect against intestinal inflammation, with improvement in the disease activity index and macroscopic damage. IA or SSZ significantly reduced myeloperoxidase activity, and also down-regulation of the gene expression of JNK1, NF-κß-p65, STAT3, and decreased levels of TNFα, IL-1ß, and increased IL-10, associated with a significant improvement of oxidative stress, in addition to a reduction in MDA and an increase of glutathione in colonic tissue. The protective effect of the extract was also confirmed in histological evaluation, showing preservation of the colonic cytoarchitecture. Immunohistochemical analysis revealed down-regulation of NF-κß-p65, iNOS, IL-17, and up-regulation of SOCs-1 and MUC-2. IA extract presents antioxidant and anti-inflammatory intestinal properties, and proved to be a potential application for preventing damage induced by DNBS.