Rita de Cássia Ribeiro Gonçalves

Introduction: Helicobacter pylori is one of the main risk factors for gastric diseases and gastric cancer due to its ability to colonize the gastric mucosa, induce chronic inflammation, and express virulence adhesins involved in bacterial persistence. Silymarin, a flavonolignan complex extracted from Silybum marianum , exhibits antioxidant, anti-inflammatory, and antimicrobial properties, suggesting potential therapeutic application against H. pylori . This study aimed to evaluate the antibacterial, anti-adhesive, anti-inflammatory, cytotoxic, and in silico interaction profiles of silymarin against H. pylori and its main adhesins BabA, SabA, and HopQ. Methods: The antibacterial activity was evaluated by broth microdilution for determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Morphological alterations were analyzed by scanning electron microscopy (SEM). Cytotoxicity was assessed by MTT assay using AGS gastric adenocarcinoma cells and RAW 264.7 macrophages. TNF-α, IL-6, and nitric oxide (NO) production were quantified in stimulated macrophages. Anti-adhesive activity was determined using FITC-labeled H. pylori adhesion assays in AGS cells. Molecular docking and physicochemical predictions were performed for the major flavonolignans present in silymarin. Results and Discussion: Silymarin exhibited MIC and MBC values of 128 μg mL ⁻¹ against H. pylori ATCC 43504, with 90.46% growth inhibition. SEM demonstrated bacterial deformation and coccoid-like morphology after treatment. Silymarin reduced TNF-α, IL-6, and NO production by 80.82%, 76.24%, and 97.50%, respectively. In adhesion assays, silymarin inhibited bacterial attachment by 25.87% ± 3.50%. Molecular docking demonstrated favorable interactions between flavonolignans and H. pylori adhesins. Isosilybinin A showed the highest affinity for BabA (fitness score = 57.75), establishing hydrogen bonds with CYS189 and SER244. Isosilychristin presented the best interaction with SabA (fitness score = 74.79), interacting with LYS152 and GLN159, while silychristin demonstrated the highest affinity for HopQ (fitness score = 66.42), interacting with SER135, ILE130, and ILE242. Silymarin demonstrated significant antibacterial, anti-inflammatory, and anti-adhesive activities against H. pylori ATCC 43504, supporting its potential as a multifunctional natural therapeutic candidate for the management of H. pylori -associated gastric diseases.

Endophytic fungi can produce metabolites with potential pharmaceutical applications through biosynthetic pathways strongly influenced by culture conditions. In this study, growth parameters (media composition and cultivation time) were systematically optimized to maximize 3-O-methylfunicone (3OMF) production by the endophytic fungus Talaromyces pinophilus J6. Chemical profiling by high-resolution mass spectrometry (HRMS) demonstrated that supplementation of potato dextrose agar (PDA) with 45% (w/w) ammonium sulfate and cultivation for 7 days significantly increased the accumulation of 3OMF compared to standard growth conditions. The optimized culture condition enabled the isolation of the target compound in sufficient yield for subsequent biological assays. The compound exhibited anti-Helicobacter pylori activity against one reference strain and three clinical isolates, with minimum inhibitory concentrations ranging from 0.165 to 0.660 mM and bactericidal concentrations up to 1.32 mM. In combination assays, 3OMF showed an additive effect with clarithromycin, indicating potential as an adjuvant compound in eradication therapies. Cytotoxicity assays of 3OMF demonstrated selective activity against gastric adenocarcinoma cells, with an IC₅₀ of 0.24 mM and selectivity index of 2.37 relative to fibroblasts. Molecular docking suggested favorable interactions between 3OMF and H. pylori β-clamp protein, suggesting a possible association with interference in bacterial DNA replication. These findings demonstrate that culture optimization is an efficient biotechnological strategy to improve 3OMF production and support the further investigation of this compound as a scaffold for anti-H. pylori applications. Graphical abstract

Colorectal cancer is one of the most prevalent malignancies worldwide, with the global incidence projected to reach approximately 3.2 million cases by 2040. Mesoporous silica nanoparticles have been extensively investigated as promising drug delivery systems for cancer therapy. Chalcones, key precursors of flavonoids, have demonstrated notable cytotoxic activity, and structural modifications involving diverse functional groups have been shown to further enhance their antitumor efficacy. Within this context, we evaluated a synthetic chalcone against colorectal cancer cells (HCT-116) using the MTT tetrazolium assay. The 2′-hydroxy-2-fluorochalcone was incorporated into MCM-41 (Mobil Composition of Matter No. 41) and into matrices functionalized with 3-aminopropyltriethoxysilane (APTES) and folic acid. The nanoparticles were synthesized and characterized by thermogravimetric analysis (TGA), elemental analysis (CHN), scanning electron microscopy (SEM), transmission electron microscopy (TEM), zeta potential, and nitrogen adsorption. Evaluate its cytotoxicity in HCT-116 cells and selectivity (IS) with L-929 cells. Molecular docking showed interactions between amino acid residues and functional groups of 2′-hydroxy-2-fluorochalcone that was very important to its cytotoxicity and activity about proteins: thymidylate synthase and vascular endothelial growth factor receptor 2 (VEGFR2). In vitro release assay was carried out to verify the release profile of the chalcone from the nanoparticles. In vitro cytotoxic tests showed that the MCM-APTES-AF-CH sample improved the IC ₅₀ [IC ₅₀ of 2.73 ± 0.96 μM (IS: 5.66)] against the HCT-116 cells. Hemolysis assay showed that the samples did not cause hemolysis in the concentrations tested. These findings indicate that MCM-APTES-AF-CH represents a promising nanocarrier platform for enhancing the anticancer efficacy of novel chalcone derivatives against colorectal cancer cells.

Paracoccidioidomycosis (PCM) is an infectious disease caused by dimorphic fungi of the Paracoccidioides genus and causes a series of discomforts in affected patients. This work aimed to evaluate the antifungal potential of synthetic chalcones against Paracoccidioides brasiliensis (Pb) and to determine in silico possible therapeutic targets. An in silico evaluation of a database of 21 synthesized chalcones was carried out based on pharmacokinetic parameters, enzymatic inhibition, Tanimoto similarity, and the prediction of the spectrum of activity by PASS (prediction of activity spectra of substances). The most viable chalcones from the previous evaluation were selected for the measurement of minimum inhibitory concentration (MIC) against Pb and for cytotoxicity assays pre- and post-metabolization using HEPG2 cells. After in silico evaluation, the compounds 4, 11, 12, 20, and 21 were selected to carry out the molecular docking and in vitro tests. In the docking studies, multiple hydrophobic and polar intermolecular interactions were observed, such as hydrogen bonds, with emphasis on compound 20 in the active site of thioredoxin, where it made 4 hydrogen bonds with the residues Gln43, Ala36, and Thr38. In vitro testing revealed antifungal activity, with the MICs ranging from 32 to 128 μg/mL. In cytotoxicity assays, the 5 compounds exhibited reduced IC50 values (5.51–14.85 μg/mL pre-metabolization and 10.48–35.4 μg/mL post-metabolization). The compounds 4, 11, 12, 20, and 21 have shown favorable predictions of pharmacokinetic characteristics and distinct actions compared to conventional medications, as well as antifungal activity with less toxicity after metabolization, making them the best candidates for further studies.

Introduction: Pharmacovigilance is a science related to the early detection and prevention of events associated with the use of drugs, such as adverse drug reactions (ADR). Active surveillance methods are increasingly being used to detect ADRs. Although the use of triggers is recommended, active Search tools for suspected ADR are scarce in the literature. Objective: To develop an active search tool for suspected ADR using triggers. Method: This methodological development study was conducted and divided into two stages: i) a literature review to build a list of triggers and the first version of the instrument; ii) content validation using the Delphi technique with the participation of professionals involved in Pharmacovigilance and Patient Safety at the institution. Instrument sessions were considered valid when the Content Validity Index (CVI) ≥ 0.80. Results: An online form was developed on the Google Forms platform containing three sections: review of medical records, patient and hospitalization data, and identification of suspected ADR. The final version has 28 questions, including 16 objective and 12 subjective. The material was evaluated by seven experts in two Delphi rounds, and the total CVI was 1. Conclusions: The instrument shows evidence of validity in terms of objective, structure, relevance, appearance, and content, representing another Pharmacovigilance tool for the hospital. It can be used by healthcare professionals from different backgrounds and is easily adaptable to the reality of other institutions. Future work should be carried out to validate the tool with the target audience.

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Aims: We synthetized 10 hydroxylated and methoxylated chalcones and evaluated them targeting MMP-9 inhibition, looking for the rate of adhesion of H. pylori in gastric cells, and then, reduction of the inflammatory response as alternative therapeutic agents for controlling the infection. Background: Helicobacter pylori is a Gram-negative bacterium that chronically infects the human stomach, a risk factor for the development of inflammatory gastrointestinal diseases, including cancer, and is classified as a group I carcinogen. It is estimated that it infects around 45% of the global population and that the persistence of the infection is related to the adhesion of the bacteria in the gastric epithelium. The progression of gastric lesions to cancer is connected to the activation of the NF-κB and MAPK pathways, especially in cagA+ strains, which are related to increased expression of MMP-9. The activation of these metalloproteinases (MMPs) contributes to the adhesion of the bacterium in gastric cells and the evolving stages of cancer, such as enabling metastasis. Due to the increasing resistance to the current therapy protocols, the search for alternative targets and candidate molecules is necessary. In this way, controlling adhesion seems to be a suitable option since it is a crucial step in the installation of the bacterium in the gastric environment. Objective: Synthetize ten hydroxylated and methoxylated chalcones. Assess their anti-H. pylori potential, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC). Evaluate their cytotoxicity in AGS cells and selectivity with L-929 cells. Analyze the results and correlate them with in silico predictions to evaluate potential anti-adhesive properties for the chalcones against H. pylori. Method: The chalcones were synthetized by Claisen-Schmidt condensation using Ba(OH)₂ or LiOH as catalysts. Predictive in silico assays in PASS Online, tanimoto similarity, ADME properties and molecular docking in MMP-9 (PDB code: 6ESM) were performed. The in vitro assays carried out were the cell viability in gastric adenocarcinoma cells (AGS) and fibroblasts (L-929) by the MMT method and anti-H. pylori, by the broth microdilution method, through the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Results: Ten chalcones were synthesized through Claisen-Schimdt condensation with yields of 10 to 52% and characterized by ¹H and ¹³C nuclear magnetic resonance (NMR) and mass spectrometry (MS). in silico data revealed the possibility of anti-H. pylori, anti-inflammatory, and MMP-9 inhibition for the chalcones. Chalcone 9 showed the best growth inhibition values for MIC and MBC, at 1 μg/mL and 2 μg/mL, respectively. Chalcones 14 and 15 likewise demonstrated excellent inhibitory results, being 2 μg/mL for both MIC and MBC. Additionally, 15 had the best MMP-9 inhibition score. Despite not corroborating the in silico findings, chalcones 10, 13, and 18 showed good cytotoxicity and the best selectivity indices. Conclusion: All compounds exhibited strong activity against H. pylori, specially 15. The predicted MMP-9 inhibition by molecular docking added to the reasonable SI and CI₅₀ values for 15 and the satisfactory reduction in the rate of survival of the bacteria, reveals that it may be acting synergically to reduce the inflammatory response and the possibilities for developing a tumor by inhibiting both bacteria and malignant cells.

Aims: This study aimed to evaluate whether pharmacotherapeutic follow-up in patients with T2DM in primary care interferes in metabolic control, cardiovascular risk, medication adherence and quality of life. Methods: A prospective clinical study was conducted at two Primary Health Units in Vitória, Espírito Santo, Brazil with 75 patients with T2DM between 40 and 70 years old. The parameters of metabolic control evaluated included fasting blood glucose, HbA1c, triglyceride/HDL-c and total cholesterol/HDL-c ratio. The cardiovascular risk was calculated based on the Framingham risk score. Adherence to medication was measured using the Brief Medication Questionnaire and quality of life was evaluated by applying the World Health Organization Quality of Life-Bref. Results: After the follow-up, there was a significant decreasing in cardiovascular risk (p=0.048) and total cholesterol/HDL-c ratio (p=0.024) and a discrete improvement in fast glucose and HbA1c levels. The quality of life scores increased for all domains (p<0.0001) and the treatment adherence also improved with 12.00% of the patients classified as low adherence in the final time, against 41.33% before the meetings. Conclusion: These results show the proposed pharmacotherapeutic follow-up influenced positively cardiovascular risk, adherence to therapy and quality of life in all domains, and, therefore, may contribute to delay the onset of the main chronic complications of the disease.

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