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Canine visceral leishmaniasis (CVL) is caused by Leishmania infantum and is endemic in many areas of southeastern Brazil. We have hypothesized that the prevalence of infection by L. infantum in dogs housed in open-admission animal shelters is beyond the range of 3.4 - 9.6% reported among dogs domiciled in similar CVL-endemic areas. Hence, this study aimed to determine the rate of L. infantum infection among dogs maintained in shelters and to investigate the epidemiology of CVL in such environments by analyzing hematological and biochemical parameters. A total of 627 dogs from 17 different shelters across the State of Minas Gerais were screened using the Dual-Path Platform test and enzyme-linked immunosorbent assay and 211 (33.6%) were found to be seropositive in both tests. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed on skin, bone marrow and lymphoid tissues of 118 seropositive dogs with inconclusive CVL clinical diagnosis and, of these, 78 (66.1%) were PCR+ for L. infantum and 7 (5.9%) were PCR+ for L. amazonensis. One dog presented a PCR-RFLP profile that was consistent with co-infection by both parasites. Leishmania amazonensis DNA was detected in skin samples of six single-infected dogs and this constitutes a novel finding. Dogs infected only with L. amazonensis were less debilitated than those infected by L. infantum, which showed typical clinical manifestations of CVL. The co-infected dog showed only mild clinical signs. The results presented herein not only support our original hypothesis but also suggest that dogs are potential reservoirs of L. amazonensis. Public health authorities should acknowledge their responsibility towards animals in collective shelters, recognize that they are potential foci of zoonotic diseases, and establish proper functioning directives to minimize transmission to humans and to other dogs., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Glioblastoma (GB) is the most common and aggressive form of primary brain tumor, in which the presence of an inflammatory environment, composed mainly by tumor-associated macrophages (TAMs), is related to its progression and development of chemoresistance. Toll-Like Receptors (TLRs) are key components of the innate immune system and their expression in both tumor and immune-associated cells may impact the cell communication in the tumor microenvironment (TME), further modeling cancer growth and response to therapy. Here, we investigated the participation of TLR4-mediated signaling as a mechanism of induced-immune escape in GB. Initially, bioinformatics analysis of public datasets revealed that TLR4 expression is lower in GB tumors when compared to astrocytomas (AST), and in a subset of TAMs. Further, we confirmed that TLR4 expression is downregulated in chemoresistant GB, as well as in macrophages co-cultured with GB cells. Additionally, TLR4 function is impaired in those cells even following stimulation with LPS, an agonist of TLR4. Finally, experiments performed in a cohort of clinical primary and metastatic brain tumors indicated that the immunostaining of TLR4 and CD45 are inversely proportional, and confirmed the low TLR4 expression in GBs. Interestingly, the cytoplasmic/nuclear pattern of TLR4 staining in cancer tissues suggests additional roles of this receptor in carcinogenesis. Overall, our data suggest the downregulation of TLR4 expression and activity as a strategy for GB-associated immune escape. Additional studies are necessary to better understand TLR4 signaling in TME in order to improve the benefits of immunotherapy based on TLR signaling., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Natural and synthetic dyes are widely used in foodstuff, medicines and cosmetics industries to enhance and/or restore the color of the final products. This study aimed to evaluate the safety of oral consumption of one carotenoids and anacardic acids-enriched extract (CAE), obtained by green extraction from cashew apple residue fibers, a byproduct of the cashew juice industry. Presenting intense yellow color, CAE could be proposed as a new natural dye. Single and repeated-dose oral toxicity (30 days) were evaluated in female Swiss mice at doses ranging from 50 to 1000 mg/kg, while (anti)mutagenic effects were evaluated in CHO-K1 cells (in vitro Cytokinesis-Block Micronucleus assay - CBMN) and in erythrocytes collected from murine bone marrow (in vivo). CAE did not induce toxic or mutagenic effects in female mice even after 30 days of treatment, regardless of the dose used. Considering cyclophosphamide (CPA)-challenged animals treated with CAE, neither antimutagenic effect was observed nor CAE increased CPA-mutagenic effects although in vitro CBMN results indicated that CAE might increase methyl methanesulfonate-induced micronuclei (MN) frequency besides promoting reduction on CPA-induced MN frequency. The obtained results suggest that CAE may be a safe source of carotenoids with potential use as industrial dye., 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 © 2020 Elsevier Inc. All rights reserved.)

The Asian invasive species Limnoperna fortunei (Dunker, 1857), known as the golden mussel, causes great economic and environmental damage due to its fixative capacity and accelerated proliferation. Molecular studies for the control of larval and adult forms are of great economic, scientific and technological interest. Here, we first report on the compositional analysis of the L. fortunei proteome obtained through shotgun analysis using LC-MS/MS. Among those 2790 proteins identified, many of them related to secretory processes and membrane receptors. Our second approach consisted in exposing the mollusc to the molluscicide niclosamide to evaluate the induced proteomic alterations. Exposure to niclosamide at 0.25 mg/L for 24 h resulted in a pronounced differential abundance of proteins when compared to those obtained when exposure was reduced to 4 h at 2.3 mg/L. In total, 342 proteins were found differentially expressed in the responsive individuals as revealed by label-free quantitative proteomics. Regarding the affected cell processes were: cell division and differentiation, cytoskeletal organization and compartment acidification (upregulated), and energy metabolism (downregulated). Our findings constitute the first inventory of the expressed proteome of the golden mussel and have the potential to contribute with a more rational proposition of molecular targets for control and monitoring of this species. SIGNIFICANCE: With the recent availability of transcriptomic and genomic data applied to L. fortunei the timing is right to interrogate its putative gene repertoire using proteomic techniques. These have the potential to validate the existence of the predicted genes, infer their relative abundance and quantify their levels as a response to environmental stressors or various agents. Here we provided an inventory of the golden mussel proteome and evaluated its response to the molluscicide niclosamide. The obtained results open new avenues for intervention aimed at its control or elimination, particularly by targeting the various cellular processes that were uncovered., Competing Interests: Declaration of Competing Interest 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 © 2020 Elsevier B.V. All rights reserved.)

Ion exchange chromatography is extensively used in the purification of biological compounds. Reliable mathematical models describing this chromatographic technique are available and can be used to improve the performance of this separation step. However, the use of synthetic mixtures for model development hampers the application of this approach with real cell extracts processed in downstream operations. This work presents an original approach for handling non-synthetic genuine mixtures of proteins, which was applied in the purification of an untagged recombinant pneumococcal surface protein A (PspA4Pro). First, evaluation was made of the efficiency of steric mass action (SMA) and modified Langmuir isotherms, which were separately used together with the equilibrium dispersive model (EDM). The data used for parameter estimation and model validation were obtained from anion exchange chromatography runs (employing Q-Sepharose FF), applied to real cell extracts produced by different cultivation strategies. Simulations showed that the models were able to describe the complex mixtures of unknown proteins. Next, the EDM and SMA approaches were used to separately describe the profile of PspA4Pro and the pool of protein impurities eluted together. The simulations showed that PspA4Pro tended to elute at the beginning of the peak, enabling the establishment of an alternative elution schedule that provided a 34% increase in the purity achieved using the anion exchange chromatography., 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 © 2019 Elsevier B.V. All rights reserved.)

In urban areas with intense vehicular traffic, particulate matter in suspension, especially the fraction of particles with ultra-fine diameter, has been regarded as the main problem of chronic diseases in susceptible populations, such as the elderly. This study aimed to determine the genotoxic effects of exposure to air pollution evaluating the association between the frequencies of micronuclei (MN) and binucleated (BN) cells in exfoliated oral mucosa cells of elderly population and exposure conditions, considering the influence of traffic and concentration of PM in different aerodynamic diameters. Traffic of passenger vehicles, heavy duty trucks and environmental concentrations of Particulate Matter were measured twice a day during 28 days before biological sampling of oral mucosa from 154 participants living in areas of distinct levels of urban traffic. Data from this study showed that the group of participants living near road traffic exhibited higher MN cell frequency, when compared to the other groups of subjects. In addition, a canonical correlation analysis between environmental and genotoxicity variables analysis revealed that high concentrations of the particulate matter were correlated with intense traffic and the genotoxicity in exfoliated oral cells., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The bis-(1,10-phenanthroline)copper(I) complex, [Cu(I)(phen) 2 ] + , was the first copper-based artificial nuclease reported in the literature. The biological and ligand-like properties of sulfonamides make them good candidates for fine-tuning the reactivity of the [Cu(phen) 2 ] motif with biomolecules. In this context, we developed three novel copper(II) complexes containing the sulfonamides sulfameter (smtrH) and sulfadimethoxine (sdmxH) and (N^N)-bidentate ligands (2,2'-biyridine or 1,10-phenantroline). The compounds were characterized by chemical and spectroscopic techniques and single-crystal X-ray crystallography. When targeting plasmid DNA, the phen-containing compounds [Cu(smtr - ) 2 (phen)] (1) and [Cu(sdmx - ) 2 (phen)] (2) demonstrated nuclease activity even in the absence of reducing agents. Addition of ascorbic acid resulted in a complete cleavage of DNA by 1 and 2 at concentrations higher than 10 μM. Experiments designed to evaluate the copper intermediates involved in the nuclease effect after reaction with ascorbic acid identified at least the [Cu(I)(N^N) 2 ] + , [Cu(I)(sulfa)(N^N)] + and [Cu(I)(sulfa) 2 ] + species. The compounds interact with DNA via groove binding and intercalation as verified by fluorescence spectroscopy, circular dichroism (CD) and molecular docking. The magnitude and preferred mode of binding are dependent on the nature of both N^N ligand and the sulfonamide. The potent nuclease activity of compounds 1 and 2 are well correlated with their antiproliferative and anti-M. tuberculosis profiles. The results presented here demonstrated the potential for further development of copper(II)-sulfonamide-(N^N) complexes as multipurpose metallodrugs., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Intestinal endoparasites in cats have received increasing attention worldwide due to the increase in the population of cats and their intimate relationship with people. However, concern about the zoonotic potential of these agents is still small. The aim of this study was evaluating the occurrence of intestinal endoparasites, the most prevalent ones, as well as the profile of affected cats. We conducted a survey of the medical records of cats treated at Small Animal Medical Clinic, in the Veterinary Hospital of the University of Veterinary Medicine and Animal Sciences-campus Botucatu, São Paulo, Brazil. Medical records between January 2011 and September 2015 were evaluated, including all cats presenting with gastrointestinal clinical disorders and had positive parasitological stool sample tests (Faust and Willis techniques) for any type of endoparasite. We evaluated the data obtained from these cats, such as sex, breed, age, diet, street access, history of parasitic diseases and season of the year. From the 1725 cats evaluated during this period, there were a total of 60 positive cases. Among these cats, 60% were males and 40% were females. Regarding breeds, 68.3% were mixed breeds, 21.7% Persian, 8.3% Siamese, and 1.7% Himalayan. The ages ranged from 1-11 years old, with an average age of 2 years and 5 months. However, cats up to 6-months old were the most frequently affected. The highest incidence of parasitized cat was for Giardia sp. (65.0%), followed by Ancylostoma spp. (40.0%), Dipylidium caninum and Toxocara cati (8.3% each). It was observed that 65% of cats presented single infections and 35% mixed infections. This survey analyzed only positive cases of gastrointestinal endoparasites, so it diverges in some respects from other results in the literature. Asymptomatic cats were not considered, but they may be carriers that eliminate parasites in the environment. However, all intestinal endoparasites isolated in cats in this study represent a potential risk of zoonotic infections, which highlights the importance of this data for the adoption of preventive measures, especially in young male cats, which were the most frequently affected in this study and in others., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The use of clays for therapeutic practice is widespread in almost all regions of the world. In this study the physicochemical and microbiological healing characteristics of a clay from Ocara, Brazil, popularly used for therapeutic uses, were analyzed. The presence of Ca, Mg, Al, Fe, and Si was observed, which initially indicated that the clay had potential for therapeutic use. The average particle size of the clay (26.3 μm) can induce the microcirculation of the skin and the XRD analysis shows that the clay is formed by kaolinite and illite, a swelling clay. During the microbiological evaluation there was the need to sterilize the clay for later incorporation into the pharmaceutical formula. The accelerated stability test at 50°C for 3 months has showed that the pharmaceutical formula remained stable with a shelf life of two years. After the stability test the wound-healing capacity of the formulation in rats was evaluated. It was observed that the treatment made with the formulation containing the Ocara clay showed the best results since the formula allowed greater formation of collagen fibers and consequent regeneration of the deep dermis after seven days of treatment and reepithelialization and continuous formation of granulation tissue at the 14th day., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Electric force microscopy (EFM) was employed in the electrical characterization of a blend of thermoplastic polyurethane (TPU) and poly(2-methoxy-5-(2'-ethyl-hexyloxy)-1,4-phenylenevinylene) (MEH-PPV) conjugated polymer. Although qualitative EFM interpretation is straightforward, its quantitative analysis always relies on approximated models. The extraction of physically reasonable parameters is normally assumed as a proof of validity of the theoretical model employed. In order to gather information about electric properties of this blend and to test the EFM technique itself, two distinct and discordant models were developed in this work to fit experimental EFM data. Even though MEH-PPV is regarded as a conductor in one model and as a dielectric in the other, both models yielded coherent and reasonable electrical properties for this blend. Such unexpected results are used to discuss the robustness or frailty of EFM in the analysis of complex materials.