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This qualitative study aimed to investigate the perception of undergraduate nursing students on health promotion in view of the use of psychoactive substances. Nine undergraduate students, attending the last disciplines of the first semester, participated in the study and were divided in two focal groups. The theoretical-methodological referential was based on Paulo Freire's theory of conscientization. Data were processed through content analysis, demonstrating that students find difficulties in planning intervention strategies in health promotion in view of the drug phenomenon, because the knowledge provided by the university has a biologist tendency, typical of the medical model, recognizing a gap between theory and practice. Nevertheless, they manage to reconstruct their knowledge and become aware of their role as future health promoters in the face of drug use and abuse. [ABSTRACT FROM AUTHOR]

The indiscriminate use of pesticides is one of the factors directly impacting bee populations. However, limited information is available on the pesticide effects on solitary bees, especially in Neotropical countries. In this scenario, this study evaluated the survival and histopathological effects caused by the neonicotinoid insecticide acetamiprid (7 ng/μL) and the fungicide azoxystrobin (10 ng/μL) in the midgut and parietal fat body of the solitary bee Centris analis. Female and male newly-emerged bees were orally exposed for 48 h to the pesticides, or alone or in combination, under laboratory conditions. The exposure to the insecticide reduced the survival of males, while the mixture reduced survival in both sexes. Acetamiprid promoted a reduction in the number of regenerative nests in the midgut, alterations of fat body cells by increasing carbohydrates in trophocytes, and reduction of oenocyte size, and increased the frequency of pericardial cells in the advanced activity stage. Both pesticides caused changes in HSP70 immunolabelling of midgut from males at the end of pesticide exposure. Comparatively, the effects on males were stronger than in females exposed to the same pesticides. Therefore, acetamiprid alone and in mixture with fungicide azoxystrobin can be harmful to males and females of Neotropical solitary bee C. analis showing lethal and sublethal effects at a concentration likely to be found in the environment., 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 © 2024 Elsevier Ltd. All rights reserved.)

The aim of this study was to select the best exopolysaccharide (EPS) producer among the Enterococcus strains to optimize, characterize, and evaluate its biological properties. Among the eleven strains, Enterococcus faecium KT990028 was selected, and the production conditions were optimized: 16.3 % (w/v) sucrose, 0.70 % (w/v) yeast extract, 8.3 % (w/v) reconstituted skimmed milk, at 38 °C in 15 h of incubation, producing 2.880 g/L of EPS. High performance anion exchange chromatography (HPAEC) analysis revealed that the molecular weight was 166.98 kDa. HPAEC, spectroscopy (FTIR), and nuclear magnetic resonance ( 1 H NMR) analyses revealed that the EPS was a heteropolysaccharide composed of galactose (37.74 %), rhamnose (19.79 %), arabinose (17.71 %), glucose (9.50 %), fucose (7.93 %), and mannose (7.33 %). Scanning electron microscopy showed a three-dimensional microstructure in the form of decompressed plates, with wrinkles, and pores. By means of dynamic light scattering (DLS), the EPS showed an average size varying from 135.25 ± 10.56 nm and 410.60 ± 45.20 nm, as the concentration was increased from 0.5 mg/mL to 2.0 mg/mL, respectively. X-ray diffraction revealed that the EPS has an amorphous and crystalline nature, while thermogravimetric analysis indicated stability up to 400 °C. The antioxidant effect (5 mg/mL) against DPPH, ABTS, OH, and O 2 was 64.50 ± 0.71 %, 47.50 ± 0.10 %, 68.36 ± 0.59 %, and 44.83 ± 0.86 %, respectively. It was also able to inhibit and biofilm disruption of Escherichia coli ATCC 25922 and Enterococcus faecalis ATCC 6057 and had an antimicrobial effect from 50 mg/mL for the strains of against Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Listeria monocytogenes ATCC 19117, Staphylococcus aureus ATCC 6538, and Enterococcus faecalis ATCC 6057. Cell cytotoxicity carried out using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay revealed that the EPS was safe and promoted the proliferation of Vero cells. Thus, the results indicated that the EPS from E. faecium KT990028 is a promising functional biopolymer for possible applications in the food and pharmaceutical fields., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024. Published by Elsevier B.V.)

Introduction: Lung cancer in never-smoker (LCINS) patients accounts for 20% of lung cancer cases, and its biology remains poorly understood, particularly in genetically admixed populations. We elucidated the molecular profile of driver genes in Brazilian LCINS., Methods: The mutational and gene fusion status of 119 lung adenocarcinomas from self-reported never-smoker patients, was assessed using targeted sequencing (NGS), nCounter, and immunohistochemistry. A panel of 46 ancestry-informative markers determined patients' genetic ancestry., Results: The most frequently mutated gene was EGFR (49.6%), followed by TP53 (39.5%), ALK (12.6%), ERBB2 (7.6%), KRAS (5.9%), PIK3CA (1.7%), and less than 1% alterations in RET, NTRK1, MET∆ex14, PDGFRA, and BRAF. Except for TP53 and PIK3CA, all other alterations were mutually exclusive. Genetic ancestry analysis revealed a predominance of European (71.1%), and a higher African ancestry was associated with TP53 mutations., Conclusion: Brazilian LCINS exhibited a similar molecular profile to other populations, except the increased ALK and TP53 alterations. Importantly, 73% of these patients have actionable alterations that are suitable for targeted treatments., (© The Author(s) 2024. Published by Oxford University Press.)

Bees are exposed to several threats, including pathogens (i.e. Nosema ceranae), pesticides and environmental contaminants. The new insecticide flupyradifurone, and the microplastics in the environment, have raised significant concerns on bee health. This study evaluated the simultaneous effects of microplastics, flupyradifurone, and N. ceranae on honey bee health, focusing on survival rates, N. ceranae replication, daily food consumption, and bee midgut histological alterations. Results showed a significant decrease in bee longevity across all treatments compared to the control, with the combination of flupyradifurone, microplastics, and N. ceranae having the most severe impact. Microplastics and flupyradifurone exposure also increased N. ceranae proliferation, especially in bees subjected to both stressors. Histological analysis revealed reduced regenerative cell nests in the midgut and changes in the nuclear matrix, indicating stress responses. Overall, the simultaneous presence of both biotic and abiotic stressors in nature can synergistically interact, leading to harmful effects on bees., 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 © 2024 Elsevier B.V. All rights reserved.)

Pesticide use is a major factor contributing to the global decline in bee populations. Sublethal effects, such as behavior alterations, are neglected in pesticide regulation for pollinators. However, these effects can bring important information to understanding the impacts of pesticides on bees' daily activities. In this study, we aimed to investigate the effects of the insecticide acetamiprid (7 ng/μL) and the fungicide azoxystrobin (10 ng/μL) on the behavior of the Neotropical solitary bee Centris analis. Female and male bees were exposed to these chemicals continuously for 48 h, followed by an additional 48 h without contaminated food, totaling 96 h of observation. We used five experimental groups: control, solvent control, insecticide, fungicide, and pesticide mixture (insecticide + fungicide). Behavioral alterations based on locomotion and light response were assessed by video tracking at 48 (end of pesticide exposure) and 96 h (end of bioassay). In addition, after recording bees at 96 h, the individuals were anesthetized for brain collection and histological evaluation of mushroom bodies to evaluate if pesticides can damage their neurons and impair the cognitive processes and responses of bees to sensory stimuli. Bees exposed to acetamiprid and pesticide mixture showed lethargic movements and impaired locomotion at 48 h. Notably, these behavioral effects were no longer evident after the bees consumed uncontaminated food for an additional 48 h, totaling 96 h from the start of pesticide exposure. Only fungicide exposure did not result in any behavioral or brain histological changes. Therefore, our study showed that acetamiprid at an estimated residual concentration, despite being classified as having low toxicity for bees, can cause significant initial locomotion disruption in solitary bees. These findings highlight the importance of considering sublethal effects in environmental risk assessment., 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 © 2024 Elsevier Ltd. All rights reserved.)

Background: Toxoplasmosis has been of public health concern due to direct associations with socioeconomic vulnerability and inadequate living conditions., Methods: Accordingly, the present study aimed to assess antibodies against T. gondii , historical reported toxoplasmosis cases and associated socio-environmental risk factors in Pinhais, a full urban area of Curitiba, currently the eighth biggest metropolitan area of Brazil. Anti- Toxoplasma gondii antibodies were assessed by an indirect immunofluorescence reaction (RIFI). Owner and dog samples were also tested by IFAT to anti- Leishmania spp. and anti- Trypanosoma cruzi antibodies., Results: Overall, 20/135 (14.8%) persons and 13/133 (9.8%) dogs from 25 different households were considered seropositive to T. gondii . All samples were seronegative to Leishmania spp. and Trypanosoma cruzi ., Conclusions: Although no significant covariates were found in the regression model, statistically associated risk factors in the bivariate analysis included no public water use ( p = 0.016) and drinking raw milk ( p = 0.041) for owners, and obesity ( p = 0.028) and tick infestation ( p = 0.03) for dogs. In addition, a spatial cluster of T. gondii seropositivity for both owners and their dogs overlapped the location of historic reported cases of human acquired, gestational and congenital toxoplasmosis. Finally, the results herein showed tick infestation as an indicator of socio-environmental risk for T. gondii exposure in the household environment, and dogs may be used as sentinels for human toxoplasmosis cases.

microRNAs (miRNAs) are small endogenous noncoding RNAs, and alterations in their expression may contribute to oncogenesis. Discovering a unique miRNA pattern holds the potential for early detection and novel treatment possibilities in cancer. This study aimed to evaluate miRNA expression in pediatric patients with gonadal germ cell tumors (GCTs), focusing on characterizing the miRNA profiles of each histological subtype and identifying a distinct histological miRNA signature for a total of 42 samples of pediatric gonadal GCTs. The analysis revealed distinct miRNA expression profiles for all histological types, regardless of the primary site. We identified specific miRNA expression signatures for each histological type, including 34 miRNAs for dysgerminomas, 13 for embryonal carcinomas, 25 for yolk sac tumors, and one for immature teratoma, compared to healthy controls. Furthermore, we identified 26 miRNAs that were commonly expressed in malignant tumors, with six miRNAs (miR-302a-3p, miR-302b-3p, miR-371a-5p, miR-372-3p, miR-373-3p, and miR-367-3p) showing significant overexpression. Notably, miR-302b-3p exhibited a significant association with all the evaluated clinical features. Our findings suggest that miRNAs have the potential to aid in the diagnosis, prognosis, and management of patients with malignant GCTs., (© 2024 The Authors. Molecular Oncology published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

Background: Considering the challenges posed by nitrogen (N) pollution and its impact on food security and sustainability, it is crucial to develop management techniques that optimize N fertilization in croplands. Our research intended to explore the potential benefits of co-inoculation with Azospirillum brasilense and Bacillus subtilis combined with N application rates on corn plants. The study focused on evaluating corn photosynthesis-related parameters, oxidative stress assay, and physiological nutrient use parameters. Focus was placed on the eventual improved capacity of plants to recover N from applied fertilizers (AFR) and enhance N use efficiency (NUE) during photosynthesis. The two-year field trial involved four seed inoculation treatments (control, A. brasilense, B. subtilis, and A. brasilense + B. subtilis) and five N application rates (0 to 240 kg N ha -1, applied as side-dress)., Results: Our results suggested that the combined effects of microbial consortia and adequate N-application rates played a crucial role in N-recovery; enhanced NUE; increased N accumulation, leaf chlorophyll index (LCI), and shoot and root growth; consequently improving corn grain yield. The integration of inoculation and adequate N rates upregulated CO 2 uptake and assimilation, transpiration, and water use efficiency, while downregulated oxidative stress., Conclusions: The results indicated that the optimum N application rate could be reduced from 240 to 175 kg N ha -1 while increasing corn yield by 5.2%. Furthermore, our findings suggest that replacing 240 by 175 kg N ha -1 of N fertilizer (-65 kg N ha -1 ) with microbial consortia would reduce CO 2 emission by 682.5 kg CO 2   -e ha -1 . Excessive N application, mainly with the presence of beneficial bacteria, can disrupt N-balance in the plant, alter soil and bacteria levels, and ultimately affect plant growth and yield. Hence, highlighting the importance of adequate N management to maximize the benefits of inoculation in agriculture and to counteract N loss from agricultural systems intensification., (© 2024. The Author(s).)

Background: Parkinson's disease (PD) causes several motor and non-motor symptoms, resulting in negative impacts on physical, mental, emotional, and social aspects of people with PD quality of life. Dance has been considered as a potential non-pharmacological intervention to improve people with PD motor and non-motor symptoms, thereby enhancing quality of life., Purpose: To analyze the self-perceive impacts of Brazilian Dance on the quality of life (physical, mental, emotional, and social) of PwPD, both before and during the COVID-19 pandemic., Methods: Fourteen participants from the "Dança & Parkinson" project were included in this qualitative study. Data collection instruments consisted of a profile and personal data sheet; assessment of accessibility to the online dance classes; Telephone Montreal Cognitive Assessment by phone call; and semi-structured interview conducted through ZOOM video call. The participants characterization data were calculated using mean, standard deviation, and percentages with the Excel Program version 2013. Qualitative data was analyzed using the Thematic Analysis technique in the Nvivo, version 8.0, qualitative analysis of text, sound, and video program., Results: The participants reported facing various challenges in dealing with PD, which negatively impact their quality of life. However, their resilience, acceptance, and dedication to treatment play an important role in coping with the issues related to the disease. Brazilian dance, both in-person before the COVID-19 pandemic and online during the pandemic, led the participants to perceive improvements in physical, mental, emotional, and social aspects of quality of life., Conclusion: The Brazilian dance appears to have a positive impact on the physical, mental, emotional, and social aspects of the participants' quality of life, both before and during the COVID-19 pandemic., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Delabary, Loch Sbeghen, Teixeira da Silva, Espedito Guzzo Júnior and Nogueira Haas.)

Background: Immune checkpoint inhibitors (ICIs) revolutionized non-small-cell lung cancer (NSCLC) treatment. However, improving patients' selection for this therapy is needed. Gene expression profile (GEP) is a promising biomarker tool. We assessed the predictive value of 48 onco-immune GEPs in an NSCLC real-world scenario., Methods: Retrospective cohort of Brazilian NSCLC patients treated with ICIs in any line. GEP was assessed in FFPE tumor tissue using the nCounter PanCancer IO360 panel, comprising 770 cancer immune genes., Results: The median age of the 135 patients was 61 years old, most male (57.8 %), history of smoking (83.6 %), ECOG-PS 0-1 (88.7 %), clinical stage IV (91.9 %) and adenocarcinoma (65.1 %). First-line ICI in 40 % of cases, alone or in combination with chemotherapy. The median follow-up was 28 months, overall survival after starting immunotherapy (post-immunotherapy survival - PIS) was 17.8 months, and real-world progression-free survival was 5.5 months. The GEP analysis was possible in 66 patients. We found that 14 different GEPs associated with PIS, namely IDO1, PD-L2, Cytotoxicity, Cytotoxic Cells, IFN Downstream, CTLA4, PD-L1, TIGIT, Lymphoid, Immunoproteasome, Exhausted CD8, IFN Gamma, TIS and APM. TIS and IFN-γ were the most significant GEPs associated with favorable outcomes. The median PIS for patients with high TIS expression was 29.2 versus 15.5 months (HR 0.42; 95 %CI; 0.17-0.67; p<0.05) for those with low expression. Similar results were observed for IFN-γ., Conclusions: The TIS (tumor inflammation signature) and IFN-γ signatures constitute predictive biomarkers to identify patients with NSCLC patients who would possibly benefit from ICI therapies., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Inc.)

Melipona scutellaris is a Brazilian stingless bee that is important for pollinating wild flora and agriculture crops. Fungicides have been widely used in agriculture, and floral residues can affect forager bees. The goal of our study was to evaluate the effects of sublethal concentrations of pyraclostrobin on the midgut ultrastructure of M. scutellaris forager workers. The bees were collected from three non-parental colonies and kept under laboratory conditions. The bees were orally exposed continuously for five days to pyraclostrobin in syrup at concentrations of 0.125 ng a.i./µL (FG1) and 0.005 ng a.i./µL (FG2). The control bees (CTL) were fed a no-fungicide sucrose solution, and the acetone solvent control bees (CAC) received a sucrose solution containing acetone. At the end of the exposure, the midguts were sampled, fixed in Karnovsky solution, and routinely processed for transmission electron microscopy. Ultrastructural analysis demonstrated that both the fungicide concentrations altered the midgut, such as cytoplasmic vacuolization (more intense in FG1), the presence of an atypical nuclear morphology, and slightly dilated mitochondrial cristae in the bees from the FG1 and FG2 groups (both more intense in FG1). Additionally, there was an alteration in the ultrastructure of the spherocrystals (FG1), which could be the result of cellular metabolism impairment and the excretion of toxic metabolites in the digestive cells as a response to fungicide exposure. The results indicate that ingested pyraclostrobin induced cytotoxic effects in the midgut of native stingless bees. These cellular ultrastructural responses of the midgut are a prelude to a reduced survival rate, as observed in previous studies.

Currently, only Apis mellifera is used in environmental regulation to evaluate the hazard of pesticides to pollinators. The low representativeness of pollinators and bee diversity in this approach may result in insufficient protection for the wild species. This scenario is intensified in tropical environments, where little is known about the effects of pesticides on solitary bees. We aimed to calculate the medium lethal dose (LD50) and medium lethal concentration (LC50) of the insecticide dimethoate in the Neotropical solitary bee Centris analis, a cavity-nesting, oil-collecting bee distributed from Brazil to Mexico. Males and females of C. analis were exposed orally to dimethoate for 48 h under laboratory conditions. Lethality was assessed every 24 h until 144 h after the beginning of the test. After the LD50 calculation, we compared the value with available LD50 values in the literature of other bee species using the species sensitivity distribution curve. In 48 h of exposure, males showed an LD50 value 1.33 times lower than females (32.78 and 43.84 ng active ingredient/bee, respectively). Centris analis was more sensitive to dimethoate than the model species A. mellifera and the solitary bee from temperate zones, Osmia lignaria. However, on a body weight basis, C. analis and A. mellifera had similar LD50 values. Ours is the first study that calculated an LD50 for a Neotropical solitary bee. Besides, the results are of crucial importance for a better understanding of the effects of pesticides on the tropical bee fauna and will help to improve the risk assessment of pesticides to bees under tropical conditions, giving attention to wild species, which are commonly neglected. Environ Toxicol Chem 2023;42:2758-2767. © 2023 SETAC., (© 2023 SETAC.)

Leptospirosis has been a neglected, widespread and reemerging zoonotic disease of global importance. The One Health holistic approach combining human, animal and environmental health has been important for control and prevention of zoonotic disease. An urban municipality in Southern Brazil with a high prevalence of leptospirosis was selected by convenience, with asymptomatic human and canine individuals serologically tested using MAT against 30 Leptospira spp. serovars. Epidemiological questionnaires were assessed along with the historical national database, with associated risk factors to dog and human seropositivity analyzed using multiple logistic regression. The spatial distribution of retrospective human leptospirosis cases was analyzed using a Kernel map and overlapped dog seropositivity and historical flooding areas, demonstrating that domestic and asymptomatic dogs may be important environmental sentinels for leptospirosis in such urban areas, even in the absence of human seropositivity and low canine seropositivity. Associated risk factors for leptospirosis in dogs and humans according to multiple regression included healthy dogs ( p = 0.02, odds ratio-OR = 0.17, confidence interval of 95%CI 0.03-0.71) with indoor access ( p = 0.059, OR = 5.12, 95%CI 1.10-37.7) and human males ( p = 0.042, OR = 2.44, 95%CI 1.08-6.11) with presenting calf pain ( p = 0.005, OR = 3.14, 95%CI 1.46-7. 14), jaundice ( p < 0.001, OR = 4.61, 95%CI 2.11-10.10) and renal failure ( p = 0.008, OR = 4.49, 95%CI 1.49-13.76). The greater the rain precipitation (above 3 mm of average annual precipitation), the higher the number of leptospirosis cases in humans. In conclusion, dog active serosurvey and rain precipitation should be systematically reported and plotted altogether as a basis for the early detection and monitoring of human risk areas. Such findings may serve as a basis for public health policies in Brazil and other endemic countries worldwide.

Salmonella is present in the poultry production chain and is a major challenge in terms of food safety and animal health. The early Salmonella detection is one of the main tools to control and prevent the transmission of this pathogen. Microbiological isolation and serotyping to identify and differentiate Salmonella serovars are laborious processes, time-consuming, and expensive. Therefore, molecular diagnostic methods can be rapid and efficient alternatives to the detection of this pathogen. Thus, the aim herein was to standardize and evaluate the use of loop-mediated isothermal amplification (LAMP) in comparison with real-time PCR (qPCR) for detection of Salmonella associated with a multiplex qPCR for simultaneous identification and differentiation of S. Enteritidis, S. Typhimurium, S. Pullorum, and S. Gallinarum. The LAMP, qPCR, and multiplex qPCR assays were comparable in specificity. The three techniques were evaluated for specificity for 16 different serovars of Salmonella and for 37 strains of the serovars of interest. The limit of detection and the efficiency of the LAMP, qPCR, and multiplex qPCR reactions were determined. The techniques were applied to 33 samples of chicken carcasses and compared to the results of conventional microbiology for validation. As results, LAMP was specific in the detection of different Salmonella serovars but presented lower limit of detection ranging from 10 1 to 10 4 CFU/reaction. In comparison, qPCR could detect less cells (10 0 to 10 2 CFU/reaction), reaching equal specificity and better repeatability in the assays. The qPCR multiplexing for identification of the different serovars also showed good specificity, with the detection threshold between entre 10 1 and 10 2 CFU/reaction. The results obtained in the analyses on poultry carcasses suggested a correspondence between the results obtained in molecular methods and in conventional microbiology. Thus, the proposed assays are promising for the diagnosis of Salmonella in poultry carcasses, already proved to be faster and more efficient than conventional diagnostics techniques, being of great interest for poultry production, animal, and public health., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Introduction and Aims: The intensive cropping system and imbalance use of chemical fertilizers to pursue high grain production and feed the fast-growing global population has disturbed agricultural sustainability and nutritional security. Understanding micronutrient fertilizer management especially zinc (Zn) through foliar application is a crucial agronomic approach that could improve agronomic biofortification of staple grain crops. The use of plant growth-promoting bacteria (PGPBs) is considered as one of the sustainable and safe strategies that could improve nutrient acquisition and uptake in edible tissues of wheat to combat Zn malnutrition and hidden hunger in humans. Therefore, the objective of this study was to evaluate the best-performing PGPB inoculants in combination with nano-Zn foliar application on the growth, grain yield, and concentration of Zn in shoots and grains, Zn use efficiencies, and estimated Zn intake under wheat cultivation in the tropical savannah of Brazil., Methods: The treatments consisted of four PGPB inoculations (without inoculation, Azospirillum brasilense, Bacillus subtilis , and Pseudomonas fluorescens , applied by seeds) and five Zn doses (0, 0.75, 1.5, 3, and 6 kg ha -1 , applied from nano ZnO in two splits by leaf)., Results: Inoculation of B. subtilis and P. fluorescens in combination with 1.5 kg ha -1 foliar nano-Zn fertilization increased the concentration of Zn, nitrogen, and phosphorus in the shoot and grain of wheat in the 2019 and 2020 cropping seasons. Shoot dry matter was increased by 5.3% and 5.4% with the inoculation of P. fluorescens , which was statistically not different from the treatments with inoculation of B. subtilis as compared to control. The grain yield of wheat was increased with increasing nano-Zn foliar application up to 5 kg Zn ha -1 with the inoculation of A. brasilense in 2019, and foliar nano-Zn up to a dose of 1.5 kg ha -1 along with the inoculation of P. fluorescens in the 2020 cropping season. The zinc partitioning index was increased with increasing nano Zn application up to 3 kg ha -1 along with the inoculation of P. fluorescens . Zinc use efficiency and applied Zn recovery were improved at low doses of nano-Zn application in combination with the inoculation of A. brasilense, B. subtilis , and P. fluorescens , respectively, as compared to control., Discussion: Therefore, inoculation with B. subtilis and P. fluorescens along with foliar nano-Zn application is considered a sustainable and environmentally safe strategy to increase nutrition, growth, productivity, and Zn biofortification of wheat in tropical savannah., Competing Interests: All the authors are aware of the manuscript publication and they have no financial, commercial, or other conflict of interest., (Copyright © 2023 Jalal, Oliveira, Fernandes, da Silva, da Costa, de Souza, Leite, Biagini, Galindo and Teixeira Filho.)

Objectives: Evatuate if Bacillus Calmette-Guérin (BCG) vaccine could be used as a tool against SARS-CoV-2 based on the concept of trained immunity., Methods: A multicenter, double-blinded, randomized clinical trial recruited health care workers (HCWs) in Brazil. The incidence rates of COVID-19, clinical manifestations, absenteeism, and adverse events among HCWs receiving BCG vaccine (Moreau or Moscow strains) or placebo were compared. BCG vaccine-mediated immune response before and after implementing specific vaccines for COVID-19 (CoronaVac or COVISHIELD) was analyzed. Cox proportional hazard and linear mixed effect modeling were used., Results: A total of 264 volunteers were included for analysis (BCG = 134 and placebo = 130). The placebo group presented a COVID-19 cumulative incidence of 0.75% vs 0.52% of BCG. The Moreau strain also presented a higher incidence rate (1.60% × 0.22%). BCG did not show a protective hazard ratio against COVID-19. In addition, the log (immunoglobulin G) level against SARS-CoV-2 presented a higher increase in the BCG group, whether or not participants had COVID-19, but also without statistical significance., Conclusion: Our results suggest that BCG has a tendency of protection against SARS-CoV-2 and higher immunoglobulin G levels than placebo. The clinical trial was registered at https://clinicaltrials.gov/ (NCT04659941)., Competing Interests: Declaration Of Competing Interest The authors have no competing interests to declare., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Leptospirosis has been described as a disease neglected worldwide. Affecting humans and animals, the disease is often related to poor environmental conditions such as lack of sanitation and presence of synanthropic rodents. Despite being considered as a One Health issue, no study has focused on comparing owner-dog seroprevalence between islands and seashore mainland. Accordingly, the present study assessed anti- Leptospira spp. antibodies by applying microscopic agglutination test (MAT) methods to Leptospira and assessing associated risk factors via univariate and multivariate logistic regression analysis of owners and their dogs in islands and seashore mainland of southern Brazil. No anti- Leptospira spp. Seropositivity was found in 330 owner serum samples, while dogs presented an overall seroprevalence of 5.9%. All seropositive dogs reacted to serogroups of Leptospira interrogans , including 66.7% of Pyrogenes, 44.4% Canicola, 22.2% Icterohaemorrhagiae, 16.7% Australis; six reacted to more than one serogroup. No association was found among seropositivity and epidemiological variables, except that neighborhood dogs were less likely to be seropositive. Although no seropositivity was observed in owners, seropositivity in dogs had the potential to indicate such species as being sentinels for environmental exposure and potential human risk of infection.

The performance of fluorescence immunostaining is physically limited by the brightness of organic dyes, whereas fluorescence labeling with multiple dyes per antibody can lead to dye self-quenching. The present work reports a methodology of antibody labeling by biotinylated zwitterionic dye-loaded polymeric nanoparticles (NPs). A rationally designed hydrophobic polymer, poly(ethyl methacrylate) bearing charged, zwitterionic and biotin groups (PEMA-ZI-biotin), enables preparation of small (14 nm) and bright fluorescent biotinylated NPs loaded with large quantities of cationic rhodamine dye with bulky hydrophobic counterion (fluorinated tetraphenylborate). The biotin exposure at the particle surface is confirmed by Förster resonance energy transfer with dye-streptavidin conjugate. Single-particle microscopy validates specific binding to biotinylated surfaces, with particle brightness 21-fold higher than quantum dot-585 (QD-585) at 550 nm excitation. The nanoimmunostaining method, which couples biotinylated antibody (cetuximab) with bright biotinylated zwitterionic NPs through streptavidin, significantly improves fluorescence imaging of target epidermal growth factor receptors (EGFR) on the cell surface compared to a dye-based labeling. Importantly, cetuximab labeled with PEMA-ZI-biotin NPs can differentiate cells with distinct expression levels of EGFR cancer marker. The developed nanoprobes can greatly amplify the signal from labeled antibodies, and thus become a useful tool in the high-sensitivity detection of disease biomarkers., (© 2023 The Authors. Small Methods published by Wiley-VCH GmbH.)

Despite being an important public health issue, particularly due to rabies, dog bites and associated risk factors have rarely been assessed by health services from a One Health perspective. Accordingly, the present study aimed to assess dog biting and associated demographic and socioeconomic risk factors in Curitiba, the eighth-largest Brazilian city with approximately 1.87 million people, based on the post-exposure prophylaxis (PEP) rabies reports between January/2010 and December/2015. The total of 45,392 PEP reports corresponded to an average annual incidence of 4.17/1000 habitants, mainly affecting white (79.9%, 4.38/1000 population), males (53.1%, 4.81/1000 population), and children aged 0-9 years (20.1%, 6.9/1000 population), with severe accidents associated with older victims ( p < 0.001) and mainly caused by dogs known to the victims. An increase of USD 100.00 in the median neighborhood income was associated with a 4.9% (95% CI: 3.8-6.1; p < 0.001) reduction in dog bites. In summary, dog biting occurrence was associated with victims' low income, gender, race/color, and age; severe accidents were associated with elderly victims. As dog bites have been described as multifactorial events involving human, animal, and environmental factors, the characteristics presented herein should be used as a basis to define mitigation, control, and prevention strategies from a One Health perspective.

(1) Background: During adolescence, there are significant changes in food consumption, such as reducing the consumption of in natura or minimally processed foods and increasing the consumption of ultra-processed foods. Thus, eating habits can influence sleep duration and, consequently, affect the quality of life of young people. This study thus aims to estimate the association of consumption of in natura or minimally processed, processed, and ultra-processed foods with sleep durations in adolescents. (2) Methods: This is a cross-sectional study including 964 adolescents (18 to 19 years old) from the 1997 to 1998 birth cohort in São Luís, Maranhão. Food consumption was assessed using the food frequency questionnaire (FFQ) and stratified based on the NOVA classification. Sleep duration was verified using accelerometry in hours. The analysis of the association between the consumption of in natura or minimally processed, processedand ultra-processed foods with sleep durations in adolescents used crude and adjusted linear regression (by gender, age, skin color, education, economic class, work, consumption of alcohol, smoking, screen time, physical activity, use of illicit drugs, anxiety, depressive symptoms, and lean and fat mass). A directed acyclic graph (DAG) was used to determine the minimum set of adjustment factors. (3) Results: Of the 964 individuals evaluated, 52.0% were female. The mean sleep duration was 6 h (± 0.95). In the crude and adjusted analyses, no association was observed between food consumption according to the degree of processing and adolescent sleep durations. (4) Conclusion: There was no association between the consumption of in natura or minimally processed, processed, and ultra-processed foods with sleep durations.

Background: Targeted and immunotherapies are currently moving toward early-stage settings for patients with non-small cell lung cancer (NSCLC). Predictive biomarkers data are scarce in this scenario. We aimed to describe the frequency of EGFR mutations and PD-L1 expression levels in early-stage non-squamous patients with NSCLC from a large, single Brazilian oncology center., Methods: We retrospectively evaluated patients with NSCLC diagnosed at an early-stage (IB to IIIA-AJCC seventh edition) at Barretos Cancer Hospital (n = 302). EGFR mutational status was assessed in FFPE tumor tissues using distinct methodologies (NGS, Cobas, or Sanger sequencing). PD-L1 expression was evaluated by immunohistochemistry (clone 22C3) and reported as Tumor Proportion Score (TPS), categorized as <1%, 1-49%, and ≥50%. We evaluated the association between EGFR mutational status and PD-L1 expression with sociodemographic and clinicopathological parameters by Fisher's test, qui-square test, and logistic regression. Survival analysis was assessed by the Kaplan-Meier method and Cox regression model., Results: EGFR mutations were detected in 17.3% (n = 48) of cases and were associated with female sex, never smokers, and longer overall and event-free survival. PD-L1 positivity was observed in 36.7% (n = 69) of cases [TPS 1-49% n = 44(23.4%); TPS ≥50% n = 25(13.3%)]. PD-L1 positivity was associated with smoking, weight loss, and higher disease stages (IIB/IIIA)., Conclusion: The frequencies of EGFR mutations and PD-L1 positivity were described for early-stage non-squamous patients with NSCLC. These results will be essential for guiding treatment strategies with the recent approvals of osimertinib and immunotherapy in the adjuvant setting., (© The Author(s) 2022. Published by Oxford University Press.)

Although toxoplasmosis has been considered among the most neglected zoonoses worldwide, no study has focused on the frequency and associated risk factors of owners and their dogs living on an island and mainland seashore areas. Accordingly, anti- Toxoplasma gondii IgG antibodies were screened by indirect fluorescent antibody test (IFAT) in owners and dogs from three oceanic islands and two nearby mainland harbor areas, with associated risk factors for toxoplasmosis assessed by univariable and multivariable logistic regression models. Overall, anti- T. gondii seropositivity was observed in 59/328 (18.0%) owners and 66/283 (23.3%) dogs, with no statistical difference between owners ( p = 0.360) and dogs ( p = 0.655) from islands and mainland areas. Consumption of local water springs ( p = 0.016; OR = 2.11) was an associated risk factor for T. gondii seropositivity, and owners with the habit of spring water intake were twice more likely seropositive ( p = 0.014; OR = 2.14). Presence of anti- T. gondii antibodies in dogs was associated with seropositive owners ( p = 0.008; OR = 2.81), household consumption of beef meat ( p = 0.042; OR = 1.7) and chicken ( p = 0.026; OR = 2.9). Despite being lower than the worldwide prevalence, toxoplasmosis seropositivity in owners and their dogs in southern Brazil was influenced by the positive owner, water source, and meat consumption, and not by inhabiting islands or seashore mainland areas, presence of dogs, cats, or both. In addition, drinking water quality should always be considered a critical risk factor for toxoplasmosis on islands.

Although leptospirosis has been described as a worldwide bacterial zoonosis primarily affecting vulnerable populations, to date no study has focused on concomitant serosurvey of homeless persons and their dogs. The aim of the present study was, to use a One Health approach to serologically assess homeless persons and their dogs in 3 major cities of south Brazil (São Paulo, Curitiba, and Foz do Iguaçu). Environmental information was obtained with an epidemiological questionnaire given to all participants. A total of 200 human and 75 dog samples were tested for anti- Leptospira spp. antibodies to thirty different serovars using the microscopic agglutination test. None of the homeless persons were positive while 5 of the 75 (6.7%) dogs were positive. Among homeless population, 89% (177 of 200) were male, 61% (122 of 200) self-declared Non-white, and 67% (134 of 200) were educated up to the 8th school grade. Lower exposure of homeless persons to Leptospira spp. in the present study when compared to other vulnerable populations (slum and low-income residents) may be result of less direct exposure as they are able to rapidly change locations in response to flooding events. In addition, these results may reflect the effectiveness of a specific healthcare service provided to people living in the streets in the 3 cities. While dogs may be used as environmental sentinels for leptospirosis, the low seropositivity results found in this report may indicate low transmission risk to homeless owners in direct daily contact with their dogs., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors. Published by Elsevier B.V.)

This study was designed to investigate the effects of Azospirillum brasilense and Bradyrhizobium sp. co-inoculation coupled with N application on soil N levels and N in plants (total N, nitrate N-NO 3 - and ammonium N-NH 4 + ), photosynthetic pigments, cowpea plant biomass and grain yield. An isotopic technique was employed to evaluate 15 N fertilizer recovery and derivation. Field trials involved two inoculations-(i) single Bradyrhizobium sp. and (ii) Bradyrhizobium sp. + A. brasilense co-inoculation-and four N fertilizer rates (0, 20, 40 and 80 kg ha -1 ). The co-inoculation of Bradyrhizobium sp. + A. brasilense increased cowpea N uptake (an increase from 10 to 14%) and grain yield (an average increase of 8%) compared to the standard inoculation with Bradyrhizobium sp. specifically derived from soil and other sources without affecting 15 N fertilizer recovery. There is no need for the supplementation of N via mineral fertilizers when A. brasilense co-inoculation is performed in a cowpea crop. However, even in the case of an NPK basal fertilization, applied N rates should remain below 20 kg N ha -1 when co-inoculation with Bradyrhizobium sp. and A. brasilense is performed.

Changes in root growth and metabolism of P in tomato cultivars are necessary in acidic soils in tropical and subtropical regions in response to P deficiency. This increase in the efficiency of phosphorus absorption by tomatoes can significantly reduce the doses of phosphate fertilizers used, as well as, possibly, the more immediate use of P fixed in the soil matrix, with favorable effects on agricultural sustainability, promoting the use of marginal areas in terms of soil fertility, and the national fertilizer economy. The tested hypothesis was that there would be no difference in the uptake and utilization of P by tomato cultivars; therefore, this study investigated the variation in the dynamics of absorption and efficiency of P-use through changes in the root, stem, leaf, gas exchange, and P-use efficiency in tomato cultivars contrasting P-absorption. The experimental design comprised a factorial scheme consisting of two cultivars that were tolerant and sensitive to P deficiency and three P concentrations (control, moderate deficiency, and severe deficiency). P limitation in the tolerant cultivar promoted high dry matter concentration (root, stem, and leaf), leaf area, root volume, nutrient translocation, rate of leaf gas exchange, and efficiency under P-deficiency stress. It was concluded from the research that the variation in the dynamics of absorption and efficiency of P use of the tolerant cultivar increased the production of roots, leaves, and leaf gas exchange under P stress conditions., (© 2022. The Author(s).)

Background: Individuals with hoarding disorder (HD) presented a persistent difficulty in detaching from objects and/or animals. Unhealthy conditions, frequently found in cases of animal HD (AHD), may favour environmental contamination and the spread of zoonotic pathogens. Despite that, only one study of zoonotic diseases in individuals with AHD and their companion animals has been conducted to date., Objectives: This study aimed to assess the seroprevalence of anti-Leptospira spp. antibodies in individuals with AHD and their dogs in a major city of Southern Brazil., Methods: Blood samples were obtained from 264 dogs (21 households) and 19 individuals with AHD (11 households) and tested by microscopic agglutination test., Results: All human samples were seronegative. Seropositivity was found in 16/264 (6.1%; CI 95% 3.3-9.6%) dogs from 11/21 (52.38%) households, with titres ranging from 100 up to 400, and Copenhageni (10/16; 62.5%) was the most frequent serovar. Surprisingly, seropositivity of hoarded dogs found herein was among the lowest reportedly observed in other dog populations of Brazil. Two epidemiological variables were significantly associated with seropositivity in dogs: the presence of cat hoarding (p = 0.004) and the report of flood occurrence in the household (p = 0.031)., Conclusions: No individuals with AHD were seropositive, and besides the lower seroprevalence of dogs, they probably had contact with Leptospira spp. at some point in their life. Since dogs can be considered potential sentinels in leptospirosis, public health programs must become aware of the risk of leptospirosis cases in households of individuals with AHD and nearby communities., (© 2022 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Pleomorphic hyalinizing angiectatic tumor (PHAT) of soft tissues is a rare, non-metastatic tumor of unknown etiology and uncertain behavior, which may recur locally. There are few reports on this condition, and due to the rarity of the disease, its lineage has not yet been fully elucidated. The present study aims to report the case of an unusual entity observed for the first time in vulval topography. A female patient, 83 years old, presented with a tumor in the vulvar region that had evolved for approximately 4 months. Magnetic resonance imaging showed an expansive perineal formation of 8.5 × 3.5 cm, and a hemivulvectomy with a flap rotation was performed. The review of the slides revealed a mesenchymal lesion without significant atypia, which was richly vascularized. In the areas of interest, the immunohistochemical (IHC) study demonstrated positivity for CD34, estrogen, and progesterone receptors; it was negative for the other tested markers. Morphological findings associated with the IHC staining panel supported the diagnosis of PHAT. The main morphological features of PHAT are clusters of ectatic vessels of different sizes that show deposits of subendothelial and intraluminal fibrin. Fusiform and pleomorphic cells randomly arranged in leaves or long fascicles intermingle these vessels. It is essential to recognize this entity and consider it among the differential diagnoses of a mesenchymal lesion, given the wide variety of entities that comprise this group of lesions., Competing Interests: Conflict of interest: None., (Copyright © 2022 The Author(s).)

COVID-19 has posed a worldwide public health challenge affecting millions of people in different countries. Rapid and efficient detection of SARS-CoV-2 is essential for pandemic control. Reverse Transcription quantitative PCR (RT-qPCR) of nasopharyngeal swabs is the gold standard method for the virus detection, but the high demand for tests has substantially increased the costs and reduced the availability of reagents, including genetic material purification kits. Thus, the present study aimed to compare two bead-based RNA extraction methods (an in-house and a commercial kit) from nasopharyngeal swabs and RT-qPCR detection of SARS-CoV-2. Twenty-five positive and five negative nasopharyngeal swab samples were subjected to extraction of nucleic acids using both methods in an automated platform. Both protocols revealed a high correlation between Cycle Quantifications (Cqs) (r = 0.99, p < 0.0001). In addition, the in-house kit was 89.5 % cheaper when compared to the mean cost of commercial RNA extraction kits. The results show that the in-house protocol is an affordable and reliable option for RNA extraction for SARS-CoV-2 detection from nasopharyngeal swabs., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Introduction: Lymph node fine needle aspiration (LN-FNA) is a minimally invasive method of evaluating lymphadenopathy. Nonetheless, its use is not widely accepted due to the lack of guidelines and a cytopathological categorisation that directly relates to management. We report our experience with LN FNA at a large Cancer Center in Latin America., Methods: We retrospectively collected cytological cases of lymph node FNA from the department of pathology at AC Camargo Cancer Center performed over a 2-year period. Data extracted included LN location, age, sex and final cytological diagnosis. Patients that had undergone neoadjuvant chemotherapy and/or cases for which the surgery specimen location was not clearly reported were excluded. For those cases with surgical reports, risk of malignancy was calculated for each diagnostic category, along with overall performance of cytology. False positive cases were reviewed to assess any possible misinterpretation or sampling errors., Results: A total of 1730 LN-FNA were distributed as follows: 62 (3.5%) non-diagnostic (ND); 1123 (64.9%) negative (NEG), 19 (1.1%) atypical (ATY), 53 (3.1%) suspicious for malignancy (SUS), and 473 (27.3%) positive (POS). Surgical reports were available for 560 cases (32.4%). Risk of malignancy (ROM) for each category was 33.3% for ND, 29.9% for NEG, 25% for ATY, 74.2% for SUS and 99.6% for POS. Overall sensitivity, specificity, negative predictive value (NPV) and positive predictive value (PPV) were 78.5%, 99.4%, 70.2% and 99.6%, respectively., Conclusion: Lymph node FNA is a very specific and accurate exam, which is reliable in the detection of lymph node metastasis and other causes of lymphadenopathy., (© 2021 John Wiley & Sons Ltd.)

The growing demand for excellent-quality coffees allied with their symbolic aestheticization that add value to the products favor the adulteration practices and consequently economic losses. So, this work proposes the suitability of NIR spectroscopy and Digital Images (from CACHAS) coupled with one-class classification methods for the non-destructive authentication of Gourmet ground roasted coffees. For this, Gourmet coffees (n = 44) were discriminated from Traditional (n = 36) and Superior (n = 10) by directly analyzing their powder without any sample preparation. Then, OC-PLS and dd-SIMCA were used to construct the models. dd-SIMCA using offset correction for NIR and RGB histogram for CACHAS achieved the best results, correctly recognizing all the 90 samples in both the training and test sets. Therefore, the proposed methodologies can be useful for both the consumers and regulatory agencies because it confirms the elevated standards of excellence of Brazilian specialty coffees, preventing fraudulent labeling, besides following the Principles of Green Analytical Chemistry., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: Gene fusions have been successfully employed as therapeutic targets for lung adenocarcinoma. However, tissue availability for molecular testing of multiples alterations is frequently unfeasible. We aimed to detect the presence of ALK, RET, and ROS1 rearrangements by a RNA-based single assay in Brazilian lung adenocarcinomas and to associate with clinicopathological features and genetic ancestry., Methods: From a FFPE series of 444 molecularly characterized lung adenocarcinomas, 253 EGFR/KRAS wild-type cases were eligible for gene rearrangement analysis. Following RNA isolation, ALK, RET, and ROS1 rearrangements were simultaneously analyzed employing the ElementsXT Custom panel (NanoString Technologies). Rearrangements were further associated with clinicopathological features and genetic ancestry of the patients., Results: The NanoString platform was performed in subset of 142 cases. Gene fusion results were conclusive for 94.4% (n=134) cases (failure rate =5.6%). ALK rearrangements were observed in 21 out of 134 cases, and associated with younger, never smokers, metastatic disease, and metastases in the central nervous system. RET and ROS1 fusions were detected in two and one out of 134 cases, respectively. Genetic ancestry was not associated with gene fusions. Overall, considering all cases for which a molecular analysis was conclusive ( EGFR/KRAS/ALK/RET/ROS1 ), ALK fusions frequency was observed in 6.5% (21/325), RET in 0.6% (2/325), and ROS1 in 0.3% (1/325)., Conclusions: This study successfully used a RNA-based single assay for the simultaneous analysis of ALK , RET , and ROS1 fusions employing routine biopsies from Brazilian patients lung adenocarcinoma allowing an extensive molecular testing for actionable rearrangements contributing to guide clinical strategies., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at http://dx.doi.org/10.21037/tlcr-20-740). The authors have no conflicts of interest to declare., (2021 Translational Lung Cancer Research. All rights reserved.)

Organotin compounds are applied in several industrial reactions and can present antifungal and antibacterial activities. Incorrect handling and storage practices of biodiesel and diesel-biodiesel blends can lead to microbial development, impacting its final quality. Concerning this problem, this work investigated the antimicrobial action of two organotin catalysts used in biodiesel production with four isolated microroorganisms (Bacillus pumilus, Pseudomonas aeruginosa, Pseudallescheria boydii, and Aureobasidium pullulans) and a pool of microorganisms (ASTM E1259 standard practice). Samples of soybean biodiesel with different concentrations of dibutyl tin dilaurate (catalyst 1) and di-n-butyl-oxo-stannane (catalyst 2) were prepared and added of mineral medium. The pool of microorganisms was inoculated and incubated at 30 °C and final biomass was weighted after 14 days. Thereafter, soybean biodiesel with catalyst 2 was used. Fungal biomass was weighted, and plate count was used to assess bacterial growth. Results show that catalysts 1 and 2 presented no inhibitory activity on the pool of microorganisms evaluated. A slight inhibitory activity was observed for B. pumilus and A. pullulans growth, but not for P. boydii, P. aeruginosa, or the pool of microorganisms. All experiment exhibited acidification higher than sterile control. Infrared analysis show less microbiological degradation products in the tin-protected fuel with ASTM inoculum. These results suggest that these tin-based catalysts show no toxic effect on native microbial population and a slight effect on some isolated microbial population in laboratory scale and for the first time shows that these organotin compounds can be employed safely as biodiesel catalyst. Graphical abstract.

Introduction: We analyzed the prevalence of non-small-cell lung cancer (NSCLC) with a programmed cell death ligand 1 (PD-L1) tumor proportion score (TPS) of ≥ 50% and compared the results with the existing data from clinical trials and databases from other countries., Materials and Methods: The Latin American Cooperative Oncology Group and Grupo Brasileiro de Oncologia Torácica performed a retrospective, cross-sectional study from August 2017 to April 2018. PD-L1 expression was collected from pathology reports from 5 laboratories in Brazil. All tests were sponsored by the pharmaceutical industry on request from the treating medical oncologist. PD-L1 expression was assessed by immunohistochemistry. The variables were summarized as absolute and relative frequencies or the median and interquartile range. Pearson's χ 2 test was used to compare the TPS categories stratified by sex, age, and histologic type. All analyses were performed with SAS, version 9.4, and were deemed statistically significant at P < .05., Results: A total of 1512 patients were included in the present study. Their median age was 66 years. Most patients were men (56.02%), and the most common histologic type was adenocarcinoma (58.04%); 109 tumors (11.31%) had EGFR mutations and 34 (3.64%) had ALK gene rearrangements. Overall, 56.54% had a PD-L1 TPS < 1%, 25.63% a TPS of 1% to 49%, and 17.83% a TPS of ≥ 50%. The factors associated with PD-L1 expression were histologic type (with adenocarcinoma samples having a greater proportion of TPS < 1%) and the laboratory that performed the test., Conclusion: The prevalence of high PD-L1 expression among the Brazilian NSCLC samples was lower than previously described in other countries, which could affect the number of patients who might be candidates for immunotherapy alone., (Copyright © 2020 Elsevier Inc. All rights reserved.)

During drier periods estuarine salinization can stimulate the colonization of marine organisms and further bioinvasion, therefore, may cause future ecological change. In this sense, we applied the Rapid Assessment Surveys (RAS) on natural and artificial hard substrates during the most intense drought period (between 2014 and 2016) since 1950, in a tropical estuary in Eastern Brazil. Through the estuary salinization the ascidian Didemnum psammatodes overgrow the dominant native taxa; this dominance taxa shift provided suitable habitat favorable for invasion of Ophiothela mirabilis across the estuary. Even the invasion starts on artificial substrates, the higher resilience of natural substrates were not enough to enable the establishment of invasive species. This baseline is the first evidence of bioinvasion influenced by climate change drought periods in tropical estuaries in South America, which is a great advance for the development of management strategies to mitigate this and future climate change scenarios., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Objectives: The aim of this study was to evaluate and compare the long-term clinical outcomes and quality of life of cats having undergone perineal urethrostomy (PU) or prepubic urethrostomy (PPU)., Methods: This clinical study followed 28 cats (PU, n = 22; PPU, n = 6) that underwent a urethrostomy, with a minimum of 1 year postoperative follow-up. Medical records, pet owner surveys and urologic laboratory tests were used for assessment. Urologic laboratory tests included serum symmetric dimethylarginine (SDMA), serum creatinine, urinalysis, urine specific gravity (USG), urine protein:creatinine (UPC) ratio and urine culture., Results: The main indications for urethrostomy were multiple catheterizations and PU stricture. The overall complication rates of PU and PPU were 31.8% and 83.3%, respectively. Recurrent urinary tract infection (UTI) and urine scald dermatitis were less frequent in PU than in PPU cats (UTI 22.7% vs 66.6%; dermatitis 4.5% vs 83.3%). Bacteriuria was present in 77.2% and 100% of PU and PPU cats, respectively. Owner satisfaction rates were excellent in 81.8% of PU and 33.3% of PPU cases., Conclusions and Relevance: A proportion of cats that underwent urethrostomy showed bacteriuria, recurrent UTIs and increased levels of SDMA. PPU is important as a salvage procedure; however, it should be limited to cases in which standard techniques for PU cannot be performed, owing to the potential for recurrent complications and lower owner satisfaction.

Background: Molecular tests can allow the rapid detection of tuberculosis (TB) and multidrug-resistant TB (MDR-TB). TB-SPRINT 59-Plex Beamedex® is a microbead-based assay developed for the simultaneous spoligotyping and detection of MDR-TB. The accuracy and cost evaluation of new assays and technologies are of great importance for their routine use in clinics and in research laboratories. The aim of this study was to evaluate the performance of TB-SPRINT at three laboratory research centers in Brazil and calculate its mean cost (MC) and activity-based costing (ABC)., Methods: TB-SPRINT data were compared with the phenotypic and genotypic profiles obtained using Bactec™ MGIT™ 960 system and Genotype® MTBDRplus, respectively., Results: Compared with MGIT, the accuracies of TB-SPRINT for the detection of rifampicin and isoniazid resistance ranged from 81 to 92% and 91.3 to 93.9%, respectively. Compared with MTBDRplus, the accuracies of TB-SPRINT for rifampicin and isoniazid were 99 and 94.2%, respectively. Moreover, the MC and ABC of TB-SPRINT were USD 127.78 and USD 109.94, respectively., Conclusion: TB-SPRINT showed good results for isoniazid and rifampicin resistance detection, but still needs improvement to achieve In Vitro Diagnostics standards.

Background: Corynebacterium striatum is an emerging multidrug-resistant (MDR) pathogen associated with immunocompromised and chronically ill patients, as well as nosocomial outbreaks. In this study, we characterized 23 MDR C. striatum isolated of bloodstream and catheter-related infections from a hospital of Rio de Janeiro., Methods: C. striatum isolates were identified by 16S rRNA and rpoB genes sequencing. The dissemination of these isolates was accomplished by pulsed-field gel electrophoresis (PFGE). All isolates were submitted to antimicrobial susceptibility testing by disk diffusion and by minimum inhibitory concentration using E-test strips methods. Antimicrobial resistance genes were detected by polymerase chain reaction. Quantitative tests were performed on four different abiotic surfaces and the ability to produce biofilm on the surface of polyurethane and silicone catheter was also demonstrated by scanning electron microscopy., Results: Eleven PFGE profiles were found. The PFGE profile I was the most frequently observed among isolates. Five different MDR profiles were found and all PFGE profile I isolates presented susceptibility only to tetracycline, vancomycin, linezolid and daptomycin. Only the multidrug-susceptible isolate did not show mutations in the quinolone-resistance determinant region (QRDR) of the gyrA gene and was negative in the search of genes encoding antibiotic resistance. The other 22 isolates were positive to resistance genes to aminoglycoside, macrolides/lincosamides and chloramphenicol and showed mutations in the QRDR of the gyrA gene. Scanning electron microscopy illustrated the ability of MDR blood isolate partaker of the epidemic clone (PFGE profile I) to produce mature biofilm on the surface of polyurethane and silicone catheter., Conclusions: Genotyping analysis by PFGE revealed the permanence of the MDR PFGE profile I in the nosocomial environment. Other new PFGE profiles emerged as etiologic agents of invasive infections. However, the MDR PFGE profile I was also found predominant among patients with hematogenic infections. The high level of multidrug resistance associated with biofilm formation capacity observed in MDR C. striatum is a case of concern.

Therapeutic monoclonal antibodies benefit to patients and the conjugation to gold nanoparticles (AuNPs) might bring additional activities to these macromolecules. However, the behavior of the conjugate will largely depend on the bulkiness of the AuNP and small sizes are moreover preferable for diffusion. Water-soluble thiolate-protected AuNPs having diameters of 2-3 nm can be synthesized with narrow polydispersity and can selectively react with incoming organic thiols via a S N 2-like mechanism. We therefore synthesized a mixed thionitrobenzoic acid- , thioaminobenzoic acid-monolayered AuNP of 2.4 nm in diameter and developed a site-selective conjugation strategy to link the AuNP to Cetuximab, an anti-epidermal growth factor receptor (EGFR) antibody used in clinic. The water-soluble 80 kDa AuNP was fully characterized and then reacted to the hinge area of Cetuximab, which was selectively reduced using mild concentration of TCEP. The conjugation proceeded smoothly and could be analyzed by polyacrylamide gel electrophoresis, indicating the formation of a 1:1 AuNP-IgG conjugate as the main product. When added to EGFR expressing glioblastoma cells, the AuNP-Cetuximab conjugate selectively bound to the cell surface receptor, inhibited EGFR autophosphorylation and entered into endosomes like Cetuximab. Altogether, we describe a simple and robust protocol for a site-directed conjugation of a thiolate-protected AuNP to Cetuximab, which could be easily monitored, thereby allowing to assess the quality of the product formation. The conjugated 2.4 nm AuNP did not majorly affect the biological behavior of Cetuximab, but provided it with the electronic properties of the AuNP. This offers the ability to detect the tagged antibody and opens application for targeted cancer radiotherapy.

In the present study, we propose an alternative technique called cytoplast fusion to improve the maturation rate and developmental competence of growing oocytes collected from early antral follicles in pigs. We examined whether the fusion of a growing oocyte with the cytoplast from a fully-grown oocyte (CFR group) could better promote maturation and developmental competence of the growing oocyte compared to germinal vesicle (GV) transfer (GVTR group). After 44 h of in vitro maturation (IVM), most growing oocytes (GR group) were still arrested at the GV stage (64.0 ± 5.1%); this number was significantly higher (P < 0.01) than that of the other groups. No matured oocyte was observed in the GR group. The maturation rate of GVTR oocytes was significantly improved (18.8 ± 3.5%) compared with that of growing oocytes. The proportion of oocytes that reached the metaphase-II (M-II) stage in the CFR group (37.8 ± 2.0%) was significantly higher (P < 0.05) than that in the GVTR group, although still lower than that in the control group (75.2 ± 4.4%). No blastocyst was derived from growing oocytes. Among in vitro fertilized GVTR oocytes, 3.0 ± 1.9% developed into blastocysts; however, this percentage showed an insignificant increase compared with the GR group. On the other hand, the percentage of CFR embryos that developed into blastocysts (12.0 ± 4.3%) was significantly higher than that of GR embryos (0.0%), although still lower than that of control embryos (27.0 ± 5.5%). Total cell number in blastocysts in the GVTR group (23.3 ± 6.9) was significantly lower (P < 0.05) than that in the control group (50.4 ± 5.0). Meanwhile, the total cell number in blastocysts derived from CFR oocytes (36.3 ± 4.8) was comparable to that of the control group. In summary, cytoplast fusion significantly improves maturation rate and developmental competence of growing oocytes compared with GV transfer.

Background: Cryptococcus gattii-induced cryptococcosis is an emerging infectious disease of humans and animals with worldwide distribution and public health importance due to its significant morbidity and mortality rate. The present study aimed to report a case of pulmonary infection by C. gattii molecular type VGII in State of São Paulo, Brazil., Case Presentation: A 5-year-old goat showing intermittent dry cough, ruminal tympany, anorexia, fever, tachycardia and tachypnea was presented for necropsy at the Veterinary Hospital of the School of Veterinary Medicine and Animal Sciences, São Paulo University, São Paulo, Brazil. Postmortem examination revealed numerous 2.0-6.0 cm diameter yellow gelatinous pulmonary masses. Tissues were evaluated by a combination of pathological, mycological, and molecular diagnostic techniques. Microscopically, pneumonia granulomatous, multifocal to coalescing, moderate, with many intralesional carminophilic yeasts was observed. The immunohistochemistry and mycological culture confirmed Cryptococcus spp. Internal transcribed spacers and orotidine monophosphate pyrophosphorylase nucleotide differentiation demonstrated that the isolate corresponds to the C. gattii VGII molecular subtype., Conclusions: To our knowledge, this is the first report of a pulmonary infection in a goat linked to C. gattii molecular type VGII in Southeastern Brazil. Our findings emphasize the need for an active surveillance program for human and animal new infections to improve the current public health policies due to expansion of the epidemiological niche of this important microorganism.

The effects of time of feed delivery (14.00 hours; 14.00 and 20.00 hours; 16.00 and 18.00 hours) on the physiological parameters, ingestive behavior, nutrient intake and production of lactating cows maintained in a semi-arid region during the hot season were evaluated. Regardless of treatment, all animals received the first feeding supply at 06.00 hours. Eight cows with an average body weight of 600 kg, average milk yield of 20 kg/day and 80 days in milk were utilized. The rectal temperature, respiratory rate and sweating rate were not affected (P > 0.05), with average values of 38.5°C, 53.8 movements/min and 104 g/m 2 /h respectively. There was no effect (P > 0.05) on the eating time (314 min/day), ruminating time (564 min/day), drinking time (61 min/day) and idle time (502 min/day). Similarly, the intake of nutrients and performance of lactating cows were not affected (P > 0.05), with average dry matter intake of 19.8 kg/day, 4% fat-corrected milk of 20.6 kg/day and milk fat concentration of 4.03 g/100 g. Since the behavior and performance have not altered, any times of feed delivery evaluated could be used to crossbred Holstein × Zebu cows maintained on a feedlot in semi-arid regions during the hot season., (© 2016 Japanese Society of Animal Science.)