Exploring Nitrogen-Functionalized Graphene Composites for Urinary Catheter Applications

Graphene has been broadly studied, particularly for the fabrication of biomedical devices, owing to its physicochemical and antimicrobial properties. In this study, the antibiofilm efficacy of graphene nanoplatelet (GNP)-based composites as coatings for urinary catheters (UCs) was investigated. GNPs were functionalized with nitrogen (N-GNP) and incorporated into a polydimethylsiloxane (PDMS) matrix. The resulting materials were characterized, and the N-GNP/PDMS composite was evaluated against single- and multi-species biofilms of Staphylococcus aureus, Pseudomonas aeruginosa, and Klebsiella pneumoniae. Both biofilm cell composition and structure were analyzed. Furthermore, the antibacterial mechanisms of action of N-GNP were explored. The N-GNP/PDMS composite showed increased hydrophobicity and roughness compared to PDMS. In single-species biofilms, this composite significantly reduced the number of S. aureus, P. aeruginosa, and K. pneumoniae cells (by 64, 41, and 29%, respectively), and decreased S. aureus biofilm culturability (by 50%). In tri-species biofilms, a 41% reduction in total cells was observed. These results are aligned with the outcomes of the biofilm structure analysis. Moreover, N-GNP caused changes in membrane permeability and triggered reactive oxygen species (ROS) synthesis in S. aureus, whereas in Gram-negative bacteria, it only induced changes in cell metabolism. Overall, the N-GNP/PDMS composite inhibited biofilm development, showing the potential of these carbon materials as coatings for UCs. (c) 2023 by the authors

Graphene-Based Coating to Mitigate Biofilm Development in Marine Environments

Due to its several economic and ecological consequences, biofouling is a widely recognized concern in the marine sector. The search for non-biocide-release antifouling coatings has been on the rise, with carbon-nanocoated surfaces showing promising activity. This work aimed to study the impact of pristine graphene nanoplatelets (GNP) on biofilm development through the representative marine bacteria Cobetia marina and to investigate the antibacterial mechanisms of action of this material. For this purpose, a flow cytometric analysis was performed and a GNP/polydimethylsiloxane (PDMS) surface containing 5 wt% GNP (G5/PDMS) was produced, characterized, and assessed regarding its biofilm mitigation potential over 42 days in controlled hydrodynamic conditions that mimic marine environments. Flow cytometry revealed membrane damage, greater metabolic activity, and endogenous reactive oxygen species (ROS) production by C. marina when exposed to GNP 5% (w/v) for 24 h. In addition, C. marina biofilms formed on G5/PDMS showed consistently lower cell count and thickness (up to 43% reductions) than PDMS. Biofilm architecture analysis indicated that mature biofilms developed on the graphene-based surface had fewer empty spaces (34% reduction) and reduced biovolume (25% reduction) compared to PDMS. Overall, the GNP-based surface inhibited C. marina biofilm development, showing promising potential as a marine antifouling coating

Vaccination of Mice with Salmonella Expressing VapA: Mucosal and Systemic Th1 Responses Provide Protection against Rhodococcus equi Infection

Conventional vaccines to prevent the pneumonia caused by Rhodococcus equi have not been successful. We have recently demonstrated that immunization with Salmonella enterica Typhimurium expressing the VapA antigen protects mice against R. equi infection. We now report that oral vaccination of mice with this recombinant strain results in high and persistent fecal levels of antigen-specific IgA, and specific proliferation of the spleen cells of immunized mice in response to the in vitro stimulation with R. equi antigen. After in vitro stimulation, spleen cells of immunized mice produce high levels of Th1 cytokines and show a prominent mRNA expression of the Th1 transcription factor T-bet, in detriment of the Th2 transcription factor GATA-3. Following R. equi challenge, a high H2O2, NO, IL-12, and IFN-γ content is detected in the organs of immunized mice. On the other hand, TNF-α and IL-4 levels are markedly lower in the organs of vaccinated mice, compared with the non-vaccinated ones. The IL-10 content and the mRNA transcription level of TGF-β are also higher in the organs of immunized mice. A greater incidence of CD4+ and CD8+ T cells and B lymphocytes is verified in vaccinated mice. However, there is no difference between vaccinated and non-vaccinated mice in terms of the frequency of CD4+CD25+Foxp3+ T cells. Finally, we show that the vaccination confers a long-term protection against R. equi infection. Altogether, these data indicate that the oral vaccination of mice with S. enterica Typhimurium expressing VapA induces specific and long-lasting humoral and cellular responses against the pathogen, which are appropriately regulated and allow tissue integrity after challenge

Produção comparativa de etanol da cana-de-açúcar, batata inglesa e beterraba por fermentação

Introduction: The search for fuels from renewable sources is extremely important for the environment, since they have less release of toxic gases during combustion. Ethanol is an example of a renewable source fuel and can be obtained through the fermentation process of yeasts of the species Saccharomyces cerevisiae and raw materials that contain sugar such as sugar cane and beet, or starchy raw materials such as potatoes. Objective: This work aims to evaluate the production of ethanol from the fermentation of sugarcane, beet and potato potatoes and to compare the ethanol content obtained by each substrate. Methods: The fermentation was carried out in conical flasks with marcerated substrates and biological yeast, in the proportion 1:10 and in the following conditions 30º C, 8 hours, under agitation. For the potato substrate, the saccharification step was added with a 1% v / v sulfuric acid solution. Then, the samples rested at room temperature for 15 hours for the measurement of ethanol, which was carried out by a densimeter and compared the alcohol content of the three fermentation processes. Results: The data obtained showed that sugarcane had an average ethanol yield of 9.83%, followed by potatoes with an average of 1.7%, and beet with an average of 1.24%. Conclusion: Thus, the sugarcane showed the best source for the production of biotenol, both because of the higher alcohol content obtained and because of the ease of production.  Introdução: A busca por combustíveis de fontes renováveis é de extrema importância para o meio ambiente, uma vez têm menor liberação de gases tóxicos durante a combustão. O etanol é um exemplo de combustível de fonte renovável e pode ser obtido pelo processo de fermentação de leveduras da espécie Saccharomyces cerevisiae e matérias-primas que possuem açúcar como cana-de-açúcar e beterraba, ou matérias-primas amiláceas como a batata. Objetivo: TEste trabalho tem por objetivo avaliar a produção de etanol a partir da fermentação do caldo de cana-de-açúcar, beterraba e batata inglesa e comparar o teor de etanol obtido por cada substrato. Métodos: Foi realizada a fermentação em erlenmeyer com substratos marcerados e fermento biológico, na proporção 1:10 e nas seguintes condições 30º C, 8 horas, sob agitação. Para o substrato batata, foi adicionada a etapa de sacarificação com uma solução de ácido sulfúrico 1 % v/v. Em seguida, as amostras repousaram em temperatura ambiente por 15 horas para a dosagem de etanol, o qual foi realizado por densímetro e comparado o teor alcóolico dos três processos de fermentação. Resultados: Os dados obtidos demonstraram que a cana-deaçúcar apresentou uma média de rendimento em etanol de 9,83%, seguido da batata com uma média de 1,7%, e a beterraba com média de 1,24%. Conclusão: Desta forma, a cana-de-açúcar mostrou melhor fonte para produção de biotenol, tanto pelo maior teor alcóolico obtido, quanto pela facilidade na produção

Production and Characterization of Graphene Oxide Surfaces against Uropathogens

Graphene and its functionalized derivatives have been increasingly applied in the biomedi-cal field, particularly in the production of antimicrobial and anti-adhesive surfaces. This study aimed to evaluate the performance of graphene oxide (GO)/polydimethylsiloxane (PDMS) composites against Staphylococcus aureus and Pseudomonas aeruginosa biofilms. GO/PDMS composites containing different GO loadings (1, 3, and 5 wt.%) were synthesized and characterized regarding their morphol-ogy, roughness, and hydrophobicity, and tested for their ability to inhibit biofilm formation under conditions that mimic urinary tract environments. Biofilm formation was assessed by determining the number of total and culturable cells. Additionally, the antibacterial mechanisms of action of GO were investigated for the tested uropathogens. Results indicated that the surfaces containing GO had greater roughness and increased hydrophobicity than PDMS. Biofilm analysis showed that the 1 wt.% GO/PDMS composite was the most effective in reducing S. aureus biofilm formation. In oppo-sition, P. aeruginosa biofilms were not inhibited by any of the synthesized composites. Furthermore, 1% (w/v) GO increased the membrane permeability, metabolic activity, and endogenous reactive oxygen species (ROS) synthesis in S. aureus. Altogether, these results suggest that GO/PDMS com-posites are promising materials for application in urinary catheters, although further investigation is required

How do Graphene Composite Surfaces Affect the Development and Structure of Marine Cyanobacterial Biofilms?

The progress of nanotechnology has prompted the development of novel marine antifouling coatings. In this study, the influence of a pristine graphene nanoplatelet (GNP)-modified surface in cyanobacterial biofilm formation was evaluated over a long-term assay using an in vitro platform which mimics the hydrodynamic conditions that prevail in real marine environments. Surface characterization by Optical Profilometry and Scanning Electron Microscopy has shown that the main difference between GNP incorporated into a commercially used epoxy resin (GNP composite) and both control surfaces (glass and epoxy resin) was related to roughness and topography, where the GNP composite had a roughness value about 1000 times higher than control surfaces. The results showed that, after 7 weeks, the GNP composite reduced the biofilm wet weight (by 44%), biofilm thickness (by 54%), biovolume (by 82%), and surface coverage (by 64%) of cyanobacterial biofilms compared to the epoxy resin. Likewise, the GNP-modified surface delayed cyanobacterial biofilm development, modulated biofilm structure to a less porous arrangement over time, and showed a higher antifouling effect at the biofilm maturation stage. Overall, this nanocomposite seems to have the potential to be used as a long-term antifouling material in marine applications. Moreover, this multifactorial study was crucial to understanding the interactions between surface properties and cyanobacterial biofilm development and architecture over time

Behavioral alterations and Fos protein immunoreactivity in brain regions of bile duct-ligated cirrhotic rats

Hepatic encephalopathy (HE) encompasses a variety of neuropsychiatric symptoms, including anxiety and psychomotor dysfunction. Although HE is a frequent complication of liver cirrhosis, the neurobiological substrates responsible for its clinical manifestations are largely unclear. In the present study, male Wistar rats were bile duct-ligated (BDL), a procedure which induces liver cirrhosis, and on the 21st day after surgery tested in the elevated plus-maze (EPM) and in an open field for anxiety and locomotor activity measurements. Analysis of Fos protein immunoreactivity (Fos-ir) was used to better understand the neurobiological alterations present in BDL animals. Plasma levels of ammonia were quantified and histopathological analysis of the livers was performed. BDL rats showed a significant decrease in the percentage of entries and time spent in the open arms of the EPM, an anxiogenic effect. These animals also presented significant decreases in Fos-ir in the lateral septal nucleus and medial amygdalar nucleus. Their ammonia plasma levels were significantly higher when compared to the sham group and the diagnosis of cirrhosis was confirmed by histopathological analysis. These results indicate that the BDL model induces anxiogenic results, possibly related to changes in the activation of anxiety-mediating circuitries and to increases in ammonia plasma levels.A Encefalopatia hepática (HE) engloba uma variedade de sintomas neuropsiquiátricos, incluindo ansiedade e disfunção psicomotora. Embora seja uma complicação frequente da cirrose hepática, os substratos neurobiológicos responsáveis por suas manifestações clínicas são em grande parte desconhecidos. No presente estudo, ratos Wistar machos foram submetidos ao procedimento cirúrgico de ligação e secção do ducto biliar (BDL; bile-duct ligation), para indução da cirrose hepática e, no 21º dia após a cirurgia, submetidos aos testes comportamentais no labirinto em cruz elevado (LCE) e campo aberto para avaliação da ansiedade e atividade locomotora. A análise da imunorreatividade à proteína Fos (Fos-ir) foi utilizada para melhor compreender as alterações neurobiológicas presentes nos animais do grupo BDL. Foi realizada a quantificação da concentração de amônia plasmática e análise histopatológica dos fígados. Os ratos do grupo BDL mostraram diminuição significativa na porcentagem de entradas e tempo gasto nos braços abertos do LCE, caracterizando efeito ansiogênico. Estes animais também apresentaram redução significativa na Fos-ir no núcleo septal lateral e núcleo medial da amígdala. A concentração plasmática de amônia foi significativamente mais elevada que a do grupo sham e o diagnóstico de cirrose foi confirmado por análise histopatológica. Estes resultados indicam que o modelo de HE induzido por BDL induz efeito ansiogênico possivelmente relacionado à ativação de circuitos mediadores da ansiedade e à hiperamonemia.Universidade Federal de São Paulo (UNIFESP) Departamento de BiociênciasUniversidade de São Paulo Instituto de Ciências Biomédicas Departamento de AnatomiaUNIFESP, Depto. de BiociênciasSciEL

Measurements of CFTR-Mediated Cl- Secretion in Human Rectal Biopsies Constitute a Robust Biomarker for Cystic Fibrosis Diagnosis and Prognosis

BACKGROUND: Cystic Fibrosis (CF) is caused by ∼1,900 mutations in the CF transmembrane conductance regulator (CFTR) gene encoding for a cAMP-regulated chloride (Cl(-)) channel expressed in several epithelia. Clinical features are dominated by respiratory symptoms, but there is variable organ involvement thus causing diagnostic dilemmas, especially for non-classic cases. METHODOLOGY/PRINCIPAL FINDINGS: To further establish measurement of CFTR function as a sensitive and robust biomarker for diagnosis and prognosis of CF, we herein assessed cholinergic and cAMP-CFTR-mediated Cl(-) secretion in 524 freshly excised rectal biopsies from 118 individuals, including patients with confirmed CF clinical diagnosis (n=51), individuals with clinical CF suspicion (n=49) and age-matched non-CF controls (n=18). Conclusive measurements were obtained for 96% of cases. Patients with "Classic CF", presenting earlier onset of symptoms, pancreatic insufficiency, severe lung disease and low Shwachman-Kulczycki scores were found to lack CFTR-mediated Cl(-) secretion (<5%). Individuals with milder CF disease presented residual CFTR-mediated Cl(-) secretion (10-57%) and non-CF controls show CFTR-mediated Cl(-) secretion ≥ 30-35% and data evidenced good correlations with various clinical parameters. Finally, comparison of these values with those in "CF suspicion" individuals allowed to confirm CF in 16/49 individuals (33%) and exclude it in 28/49 (57%). Statistical discriminant analyses showed that colonic measurements of CFTR-mediated Cl(-) secretion are the best discriminator among Classic/Non-Classic CF and non-CF groups. CONCLUSIONS/SIGNIFICANCE: Determination of CFTR-mediated Cl(-) secretion in rectal biopsies is demonstrated here to be a sensitive, reproducible and robust predictive biomarker for the diagnosis and prognosis of CF. The method also has very high potential for (pre-)clinical trials of CFTR-modulator therapies.This work was supported by grants TargetScreen2 (EU/FP6/LSH/2005/037365), PIC/IC/83103/2007; PTDC/MAT/118335/2010; PEstOE/BIA/UI4046/2011 (to BioFIG) and PEstOE/MAT/UI0006/2011 (to CEAUL) from FCT (Portugal); and FAPESP (SPRF, Brazil), CNPq (40.8924/2006/3, Brazil) and Mukoviszidose e.V. S02/10 (Germany). MS and IU are recipients of SFRH/BD/35936/2007 and SFRH/BD/69180/2010 PhD fellowships (FCT, Portugal), respectively. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Educação Ambiental envolvendo o descarte de pilhas e baterias: uma ação do PIBID de Química na comunidade do Salobrinho

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