Identification of R-Spondin Gene Signature Predictive of Metastatic Progression in BRAFV600E-Positive Papillary Thyroid Cancer

Papillary thyroid carcinoma (PTC) is the most common malignancy of the thyroid gland and early stages are curable. However, a subset of PTCs shows an unusually aggressive phenotype with extensive lymph node metastasis and higher incidence of locoregional recurrence. In this study, we investigated a large cohort of PTC cases with an unusual aggressive phenotype using a high-throughput RNA sequencing (RNA-Seq) to identify differentially regulated genes associated with metastatic PTC. All metastatic PTC with mutated BRAF (V600E) but not BRAF wild-type expressed an up-regulation of R-Spondin Protein 4 (RSPO4) concomitant with an upregulation of genes involved in focal adhesion and cell-extracellular matrix signaling. Further immunohistochemistry validation confirmed the upregulation of these target genes in metastatic PTC cases. Preclinical studies using established PTC cell lines support that RSPO4 overexpression is associated with BRAF V600E mutation and is a critical upstream event that promote activation of kinases of focal adhesion signaling known to drive cancer cell locomotion and invasion. This finding opens up the potential of co-targeting B-Raf, RSPO and focal adhesion proteins as a pharmacological approach for aggressive BRAF V600E PTC

O estudo do Direito Internacional sob uma nova perspectiva: nossa experiência na Philip C. Jessup International Moot Court Competition

O Estudo do Direito Internacional sob uma Nova Perspectiva: Nossa Experiência na Philip C. Jessup International Moot Court Competitio

Antimicrobial effect of Morita-Baylis-Hillman adducts against oral pathogens and cellular viability in human leukocytes / Efeito antimicrobiano de adutos de Morita-Baylis-Hillman contra patógenos orais e viabilidade celular em leucócitos de humanos

The aim of this work was to evaluate Morita-Baylis-Hillman adduct (MBHA) antimicrobial effect against oral pathogens and related cell viability in human leukocytes. Minimum inhibitory concentration (MIC) was determined using microdilution method. Cell viability was assessed in human peripheral blood mononuclear cells (PBMCs) using the resazurin assay. For S. aureus (ATCC 15656) and S. mutans (UA 159) MIC values of 2,000 ?g/mL were reported for the A1 Morita-Baylis-Hillman adducts. The MICs of the A2 and A3 adducts were not found for the bacterial strains. MIC values for the A1 adduct was 125 ?g/mL, A2 1,000 ?g/mL and A3 to 15.6?g/mL against the C. albicans strain (ATCC 11006). PBMCs showed cell viability greater than 50 % when in contact with concentrations 10x higher than MIC of MBHA. It is concluded that MBHA A1, A2 and A3 present potential antimicrobial effects against C. albicans without presenting substantial cytotoxic effects in human cells, highlighting adduct A3 for future therapeutic applications

Síntese, caracterização, citotoxicidade e imunomodulação de vidros bioativos dopados com prata em leucócitos humanos

Bioactive glasses (BG) has application in tissue engineering for bone regeneration, coating for implants and stimulatory of wound healing. BG can be conjugated to antimicrobial ions like silver (Ag) giving it antibacterial activity. Once the immunomodulatory activity of bioactive glasses doped with silver was not investigated before, the aim of this work was to evaluate the cytotoxic and immunomodulatory effect of BG and silver-doped bioactive silica (BGAg). BG and BGAg samples belonging to the system 58SiO2∙(36-x)CaO·6P2O5·xAg2O, where x = 0 and 1 mol%, respectively, were synthesized via sol–gel method, and characterized by Scanning Electron Microscopy (SEM), Atomic Force Microscopy (AFM), Surface-enhanced Raman Spectroscopy (Raman-SERS), Differential Thermal Analysis (DTA), Thermal Gravimetric Analysis (TGA) and X-ray diffraction (XRD). Cytotoxicity, cytokine modulation and oxidative stress were performed in human peripheral blood mononuclear cells (PBMCs) and polymorphonuclear neutrophils (PMNs) cultures. Microscopy characterization showed particles ≤74 μm with irregular and porous surface. RAMAN-SERS analyses confirmed silver inclusion within BGAg samples. PBMC viability in presence of BG or BGAg was dose-dependent. Samples of the BGAg presented high toxicity (LC50 = 0.005%) relative to BG (LC50 = 0.106%) against PBMCs. The glasses did not changed cytokines and Reactive oxygen species (ROS) basal secretion profiles. Therefore, BG samples decrease ROS’s levels when coincubated with positive control. Overall, Ag-modified glass particles had impact on the morphology or physical properties, although ion incorporation has increased its cytotoxicity.NenhumaO vidro bioativo (BG) possui aplicações no campo da engenharia tecidual, como o preenchimento para regeneração óssea, estimulante da regeneração e revestimento de implantes. O BG pode ser conjugado à prata (Ag), o que pode lhe conferir propriedades antimicrobianas. Considerando que a atividade imunomoduladora do BG associado a prata (BGAg) não tenha sido avaliada, o objetivo deste trabalho foi avaliar a citotoxicidade e atividade imunomoduladora do BG e do BGAg. Amostras pertencentes ao sistema 58SiO2∙(36-x)CaO·6P2O5·xAg2O, onde x = 0 ou 1 mol%, respectivamente, foram sintetizadas pelo método sol–gel, e caracterizadas por Microscopia Eletrônica de Varredura (SEM), Microscopia de Força Atômica (AFM), Análise Termogravimétrica (TGA), Difração de raios X (XRD) e Espectroscopia Raman Amplificada por Superfície (Raman-SERS). A citotoxicidade foi avaliada em monócitos (PBMCs) e polimorfonucleares (PMNs) humanos de sangue periférico. A caracterização microscópica mostrou partículas ≤74 μm com aspecto irregular e superfície porosa. As análises RAMAN-SERS confirmaram a inclusão de prata nas micropartículas de sílica. A viabilidade dos PBMCs na presença do BG e do BGAg mostrou-se ser dependente da dose. As amostras do BGAg apresentaram maior toxicidade (LC50 = 0,005%) quando comparadas com as amostras do BG (LC50 = 0,106%). Os vidros bioativos não alteraram o perfil basal de liberação de citocinas e espécies reativas de oxigênio (ROS), embora as amostras BG tenham sido capazes de reduzir os níveis de ROS induzidos por zymozan. Portanto, as partículas de vidro bioativo dopado com prata não difere na morfologia ou propriedades físicas, apesar de que a incorporação dessa modificação ter aumentado a citotoxicidade desse material, com potencial imunomodulador de resposta imune inata

In Vitro Effect of Cinnamomum zeylanicum Blume Essential Oil on Candida spp. Involved in Oral Infections

The present study demonstrates the antifungal potential of chemically characterized essential oil (EO) of Cinnamomum zeylanicum Blume on Candida spp. biofilm and establishes its mode of action, effect on fungal growth kinetics, and cytotoxicity to human cells. The minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) values varied from 62.5 to 1,000 μg/mL, and the effect seems to be due to interference with cell wall biosynthesis. The kinetics assay showed that EO at MICx2 (500 μg/mL) induced a significant (p 0.05) in the viability of human red blood cells at concentrations up to 1,000 μg/mL. Phytochemical analysis identified eugenol as the main component (68.96%) of the EO. C. zeylanicum Blume EO shows antifungal activity, action on the yeast cell wall, and a deleterious effect on Candida spp. biofilms. This natural product did not show evidence of cytotoxicity toward human cells

Anticandida and antibiofilm activities of extract from Schinopsis brasiliensis Engl. against Candida spp.

Abstract The pathogenic nature of infections caused by Candida spp. underscores the necessity for novel therapeutic agents. Extracts of Schinopsis brasilienses Engl are \ a promising source of agents with antifungal effects. This study aimed to assess the antifungal potential of the leaf extract of S. brasilienses. The antifungal activity was evaluated by determining the minimum inhibitory concentrations and fungicide concentrations (MIC and MFC). The antibiofilm potential was assessed by counting colony-forming units/mL. The study examined the inhibition kinetics of fungal growth and potential synergism between gallic acid or the extract and nystatin using the Checkerboard method. Cytotoxicity was evaluated through the MTT assay. The extract exhibited antifungal effect against all tested strains, with MIC and MFC ranging from 31.25–250 μg/mL. Gallic acid, the main isolated compound, displayed a MIC of 2000 μg/mL. The extract of S. brasilienses at 31.25 μg/mL inhibited the formation of biofilm by C. albicans and significantly reduced the mass of mature biofilm after 24 and 48 h (p < 0. 05). At a concentration of 125 μg/mL, the extract demonstrated significant inhibition of fungal growth after 6 hours. The combination of gallic acid or extract with nystatin did not exhibit synergistic or antagonistic effect. Furthermore, the extract did not induce cytotoxicity to a human cell line. The extract of S. brasiliensis demonstrates antifungal activity against Candida, generally exhibiting fungicidal action and capacity to inhibit biofilm formation as well as reduce mature biofilms. Additionally, the extract showed low cytotoxicity to human cells

Antibiofilm activity and mechanism of action of the disinfectant chloramine T on candida spp., and its toxicity against human cells

We evaluated the antifungal and anti-biofilm activity, mechanism of action and cytotoxicity of chloramine T trihydrate (CAT) against Candida spp. The Minimum Inhibitory and Fungicidal Concentrations (MIC/MFC) of CAT were determined. Changes in CAT-treated C. albicans growth kinetics and micromorphology were evaluated, as well as the mechanism of action, and its effects on biofilm. Cytotoxicity was assessed by the hemolysis method. The data were analyzed by inferential statistics (p ≤ 0.05). CAT showed antifungal activity against all strains, with MIC values ranging between 1.38 and 5.54 mmol/L (MIC75%: 2.77 mmol/L). CAT demonstrated an immediate and sustained action on C. albicans growth kinetics, particularly at 2 × MIC. This compound likely acts on the cell wall and membrane permeability simultaneously and was found to cause changes in C. albicans micromorphology. Tha antibiofilm activity of CAT was similar to that of sodium hypochlorite (p > 0.05) against mature biofilms. CAT was more effective than NaOCl in reducing mature biofilm upon 1-min exposure at 2 × MIC (24 h) and 4 × MIC (48 h) (p < 0.05). Toxicological analysis revealed that CAT had hemolytic activity between 61 and 67.7% as compared to 100% by NaOCl. CAT has antifungal and anti-biofilm properties, probably acting on both cell wall and membrane permeability, and showed low toxicity in vitro229CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQSem informaçã

Antibiofilm Activity and Mechanism of Action of the Disinfectant Chloramine T on Candida spp., and Its Toxicity against Human Cells

We evaluated the antifungal and anti-biofilm activity, mechanism of action and cytotoxicity of chloramine T trihydrate (CAT) against Candida spp. The Minimum Inhibitory and Fungicidal Concentrations (MIC/MFC) of CAT were determined. Changes in CAT-treated C. albicans growth kinetics and micromorphology were evaluated, as well as the mechanism of action, and its effects on biofilm. Cytotoxicity was assessed by the hemolysis method. The data were analyzed by inferential statistics (p ≤ 0.05). CAT showed antifungal activity against all strains, with MIC values ranging between 1.38 and 5.54 mmol/L (MIC75%: 2.77 mmol/L). CAT demonstrated an immediate and sustained action on C. albicans growth kinetics, particularly at 2 × MIC. This compound likely acts on the cell wall and membrane permeability simultaneously and was found to cause changes in C. albicans micromorphology. Tha antibiofilm activity of CAT was similar to that of sodium hypochlorite (p &gt; 0.05) against mature biofilms. CAT was more effective than NaOCl in reducing mature biofilm upon 1-min exposure at 2 × MIC (24 h) and 4 × MIC (48 h) (p &lt; 0.05). Toxicological analysis revealed that CAT had hemolytic activity between 61 and 67.7% as compared to 100% by NaOCl. CAT has antifungal and anti-biofilm properties, probably acting on both cell wall and membrane permeability, and showed low toxicity in vitro

O estudo do Direito Internacional sob uma nova perspectiva: nossa experiência na Philip C. Jessup International Moot Court Competition

- DOI: 10.5102/rdi.v17i2.729