Decidual production of interferon lambda in response to ZIKV persistence: Clinical evidence and in vitro modelling.

Zika virus (ZIKV) infections during pregnancy can result in Congenital Zika Syndrome (CZS), a range of severe neurological outcomes in fetuses that primarily occur during early gestational stages possibly due to placental damage. Although some placentas can maintain ZIKV persistence for weeks or months after the initial infection and diagnosis, the impact of this viral persistence is still unknown. Here, we aimed to investigate the immunological repercussion of ZIKV persistence in term placentas. As such, term placentas from 64 pregnant women diagnosed with Zika in different gestational periods were analyzed by ZIKV RT-qPCR, examination of decidua and placental villous histopathology, and expression of inflammation-related genes and IFNL1-4. Subsequently, we explored primary cultures of term decidual Extravillous Trophoblasts (EVTs) and Term Chorionic Villi (TCV) explants, as in vitro models to access the immunological consequences of placental ZIKV infection. Placenta from CZS cases presented low IFNL1-4 expression, evidencing the critical protective role of theses cytokines in the clinical outcome. Term placentas cleared for ZIKV showed increased levels of IFNL1, 3, and 4, whether viral persistence was related with a proinflammatory profile. Conversely, upon ZIKV persistence placentas with decidual inflammation showed high IFNL1-4 levels. In vitro experiments showed that term EVTs are more permissive, and secreted higher levels of IFN-α2 and IFN-λ1 compared to TCV explants. The results suggest that, upon ZIKV persistence, the maternal-skewed decidua contributes to placental inflammatory and antiviral signature, through chronic deciduitis and IFNL upregulation. Although further studies are needed to elucidate the mechanisms underlying the decidual responses against ZIKV. Hence, this study presents unique insights and valuable in vitro models for evaluating the immunological landscape of placentas upon ZIKV persistence.

Integrated control of neglected tropical diseases in Brazil: document review of a national campaign in light of WHO recommendations.

Objective: To describe the results of a national campaign aimed at the integrated control of neglected tropical diseases in Brazil in light of the World Health Organization (WHO) official documentation related to the integration of strategies for the prevention, control, and elimination or eradication of neglected tropical diseases. Methods: A document review that included official WHO documents published between 2007 and 2020 and campaign results extracted from the official technical report produced by the Brazilian Ministry of Health. Results: The integrated control of neglected tropical diseases was gradually incorporated in the WHO documentation over time. Preventive chemotherapy through mass drug administration, intensified case management, and integrated vector management were extensively recommended as strategies for integrated control. The Brazilian campaign was carried out in four iterations between 2013 and 2017. Children aged 5 to 14 years enrolled in municipal public schools nationwide were targeted. In summary, a total of 1 074 and 73 522 new cases of leprosy and trachoma, respectively, were detected. Nearly 18 million doses of preventive chemotherapy for soil-transmitted helminthiasis were administered. More than 700 cases of schistosomiasis were diagnosed and treated. Conclusions: The integrated strategies implemented in Brazil throughout the campaign generated results aligned with the WHO recommendations for the control of neglected tropical diseases, especially those regarding mass drug administration, active case detection, and intensified case management. Therefore, the continuity of the campaign with adequate evaluation tools must be encouraged as a constant public health policy in the Brazilian government agenda.

Impact of socioeconomic status on the knowledge, attitudes, and practices about visceral leishmaniasis among dog owners.

INTRODUCTION: This cross-sectional study evaluated the impact of socioeconomic status on the knowledge, attitudes, and practices (KAP) concerning zoonotic visceral leishmaniasis (VL) among dog owners from the municipality of Rondonópolis-a highly endemic area for the disease in Central-Western Brazil. METHODOLOGY: Data were collected between 2016 and 2017 during a household survey. A probabilistic sample of 404 dog owners were interviewed assessing sociodemographic characteristics, previous occurrence of VL cases, and KAP about human VL, vector, and canine VL. Responses regarding KAP were compared among social classes, which are indicators of socioeconomic status. Correct/appropriate answers were scored, and a multivariate Poisson regression analysis evaluated the impact of social class on scoring. RESULTS: The overall KAP regarding VL was limited. Dog owners from higher social classes differed from those of the lower classes regarding the recognition of abdominal distension (p = 0.026) and skin lesions (p < 0.001) as clinical manifestations of human and canine VL, respectively, knowledge of VL transmission (p = 0.020), use of topical repellents (p < 0.001), use of insecticide-impregnated collars (p = 0.003), and previous attempts of treatment for canine VL (p = 0.005). Higher scores were associated with the upper social classes (IRR = 1.18; CI = 1.08-1.29) adjusted by the age (IRR = 1.13; CI = 1.04-1.24) and the previous occurrence of human (IRR = 1.21; CI = 1.07-1.36) and canine (IRR = 1.25; CI = 1.14-1.36) VL in the household/neighbourhood of the respondents. CONCLUSIONS: Improved KAP concerning VL was associated with better socioeconomic status of dog owners.

The clinical spectrum and immunopathological mechanisms underlying ZIKV-induced neurological manifestations.

Since the 2015 to 2016 outbreak in America, Zika virus (ZIKV) infected almost 900,000 patients. This international public health emergency was mainly associated with a significant increase in the number of newborns with congenital microcephaly and abnormal neurologic development, known as congenital Zika syndrome (CZS). Furthermore, Guillain-Barré syndrome (GBS), a neuroimmune disorder of adults, has also been associated with ZIKV infection. Currently, the number of ZIKV-infected patients has decreased, and most of the cases recently reported present as a mild and self-limiting febrile illness. However, based on its natural history of a typical example of reemerging pathogen and the lack of specific therapeutic options against ZIKV infection, new outbreaks can occur worldwide, demanding the attention of researchers and government authorities. Here, we discuss the clinical spectrum and immunopathological mechanisms underlying ZIKV-induced neurological manifestations. Several studies have confirmed the tropism of ZIKV for neural progenitor stem cells by demonstrating the presence of ZIKV in the central nervous system (CNS) during fetal development, eliciting a deleterious inflammatory response that compromises neurogenesis and brain formation. Of note, while the neuropathology of CZS can be due to a direct viral neuropathic effect, adults may develop neuroimmune manifestations such as GBS due to poorly understood mechanisms. Antiganglioside autoantibodies have been detected in multiple patients with ZIKV infection-associated GBS, suggesting a molecular mimicry. However, further additional immunopathological mechanisms remain to be uncovered, paving the way for new therapeutic strategies.

Combining surface-sensitive microscopies for analysis of biological tissues after neural device implantation.

In order to address the complexity of chemical analysis of biological systems, time-of-flight secondary ion mass spectrometry (ToF-SIMS), x-ray photoelectron spectroscopy (XPS), and x-ray photoemission electron microscopy (XPEEM) were used for combined surface imaging of a biological tissue formed around a surface neural device after implantation on a nonhuman primate brain. Results show patterns on biological tissue based on extracellular matrix (ECM) and phospholipid membrane (PM) molecular fragments, which were contrasted through principal component analysis of ToF-SIMS negative spectrum. This chemical differentiation may indicate severe inflammation on tissue with an early case of necrosis. Quantification of the elemental composition and the chemical bonding states on both ECM-rich and PM-rich features was possible through XPS analysis from survey and high-resolution spectra, respectively. Variable amounts of carbon (68%-80.5%), nitrogen (10%-2.4%), and oxygen (20.8%-16.5%) were detected on the surface of the biological tissue. Chlorine, phosphorous sodium, and sulfur were also identified in lower extends. Besides that, analysis of the C 1s high-resolution spectra for the same two regions (ECM and PM ones) showed that a compromise between C-C (41.8 at. %) and C-N/C-O (35.6 at. %) amounts may indicate a strong presence of amino acids and proteoglycans on the ECM fragment-rich region, while the great amount of C-C (70.1 at. %) on the PM fragment-rich region is attributed to the large chains of fatty acids connected to phospholipid molecules. The micrometer-scale imaging of these chemical states on tissue was accomplished through XPEEM analysis. The C-C presence was found uniformly distributed across the entire analyzed area, while C-N/C-O and C=O were in two distinct regions. The combination of ToF-SIMS, XPS, and XPEEM is shown here as a powerful, noninvasive approach to map out elemental and chemical properties of biological tissues, i.e., identification of chemically distinct regions, followed by quantification of the surface chemical composition in each distinct region.

Mitochondrial Respiratory Inhibition Promoted by Pyraclostrobin in Fungi is Also Observed in Honey Bees.

There is no use restriction associated with bees for many fungicides used in agriculture; however, this does not always mean that these pesticides are harmless for these nontarget organisms. We investigated whether the fungicide pyraclostrobin, which acts on fungal mitochondria, also negatively affects honey bee mitochondrial bioenergetics. Honey bees were collected from 5 hives and anesthetized at 4 °C. The thoraces were separated, and mitochondria were isolated by grinding, filtering, and differential centrifugation. An aliquot of 0.5 mg of mitochondrial proteins was added to 0.5 mL of a standard reaction medium with 4 mM succinate (complex II substrate) plus 50 nM rotenone (complex I inhibitor), and mitochondrial respiration was measured at 30 °C using a Clark-type oxygen electrode. Mitochondrial membrane potential was determined spectrofluorimetrically using safranin O as a probe, and adenosine triphosphate (ATP) synthesis was determined by chemiluminescence. Pyraclostrobin at 0 to 50 µM was tested on the mitochondrial preparations, with 3 repetitions. Pyraclostrobin inhibited mitochondrial respiration in a dose-dependent manner at concentrations of 10 µM and above, demonstrating typical inhibition of oxidative phosphorylation. Pyraclostrobin also promoted a decline in the mitochondrial membrane potential at doses of 5 µM and above and in ATP synthesis at 15 µM and above. We conclude that pyraclostrobin interferes with honey bee mitochondrial function, which is especially critical for the energy-demanding flight activity of foraging bees. Environ Toxicol Chem 2020;39:1267-1272. © 2020 SETAC.

Hyperendemicity, heterogeneity and spatial overlap of leprosy and cutaneous leishmaniasis in the southern Amazon region of Brazil.

Neglected tropical diseases characterized by skin lesions are highly endemic in the state of Mato Grosso, Brazil. We analyzed the spatial distribution of leprosy and Cutaneous Leishmaniasis (CL) and identified the degree of overlap in their distribution. All new cases of leprosy and CL reported between 2008 and 2017 through the national reporting system were included in the study. Scan statistics together with univariate Global and Local Moran's I were employed to identify clusters and spatial autocorrelation for each disease, with the spatial correlation between leprosy and CL measured by bivariate Global and Local Moran's I. Finally, we evaluated the demographic characteristics of the patients. The number of leprosy (N = 28,204) and CL (N = 24,771) cases in Mato Grosso and the highly smoothed detection coefficients indicated hyperendemicity and spatial distribution heterogeneity. Scan statistics demonstrated overlap of high-risk clusters for leprosy (RR = 2.0; P <0.001) and CL (RR = 4.0; P <0.001) in the North and Northeast mesoregions. Global Moran's I revealed a spatial autocorrelation for leprosy (0.228; P = 0.001) and CL (0.311; P = 0.001) and a correlation between them (0.164; P = 0.001). Both diseases were found to be concentrated in urban areas among men aged 31-60 years, of brown-skinned ethnicity and with a low educational level. Our findings indicate a need for developing integrated and spatially as well as socio-demographically targeted public health policies.

Different aspects of platelet evaluation in dengue: Measurement of circulating mediators, ability to interact with the virus, the degree of activation and quantification of intraplatelet protein content.

Platelets play a role in hemostasis, coagulation, angiogenesis, inflammation and immune response is one of the most affected cells in dengue. Here we describe some aspects of platelets by observing their specific circulating mediators, the ability to interact with the virus and morphological consequences of this interaction, activation markers and intraplatelet protein contents in dengue. We conducted this study using dengue-patients as well as healthy donors. Immunoenzymatic assay, flow cytometry, transmission electron microscopy and intraplatelet proteins expression assays were carried out. Briefly, we found an increase in sCD62L, NO or TBX2 ratio in platelet count, mostly in patients with the worse clinical outcome. After in vitro DENV infection or during natural infection, platelets underwent morphological alteration with increased expression of platelet activation markers, particularly in natural infections. Analysis of intraplatelet protein contents revealed different angiogenic and inflammatory profiles, maintaining or not extracellular matrix integrity between DF and DFWS patients. Thus, platelets are frequently affected by dengue, either by altering their own functionality, as "carrier" of the virus, or as an antiviral and mediator-secreting effector cell. Thus, strategies aimed at recovering platelet amounts in dengue seem to be essential for a better clinical outcome of the patients.

Is zebrafish (Danio rerio) a tool for human-like metabolism study?

One of the greatest challenges in anti-doping science is the large number of substances available and the difficulty in finding the best analytical targets to detect their misuse. Therefore, metabolism studies involving prohibited substances are fundamental. However, metabolism studies in humans could face an important ethical bottleneck, especially for non-approved substances. An emerging model for metabolism assessment is the zebrafish, due to its genetic similarities with humans. In the present study, the ability of adult zebrafish to produce metabolites of sibutramine and stanozolol, substances with a well-known metabolism that are widely used as doping agents in sports, was evaluated. They represent 2 of the most abused classes of doping agents, namely, stimulants and anabolic steroids. These are classes that have been receiving attention because of the upsurge of synthetic analogues, for which the side effects in humans have not been assessed. The samples collected from the zebrafish tank water were hydrolysed, extracted by solid-phase extraction, and analysed by liquid chromatography with high resolution mass spectrometry (LC-HRMS). Adult zebrafish could produce several sibutramine and stanozolol metabolites, including demethylated, hydroxylated, dehydroxylated, and reduced derivatives, all of which have already been detected in human urine. This study demonstrates that adult zebrafish can absorb, oxidise, and excrete several metabolites in a manner similar to humans. Therefore, adult zebrafish seem to be a very promising tool to study human-like metabolism when aiming to find analytical targets for doping control. Copyright © 2017 John Wiley & Sons, Ltd.