https://repositorio.ufms.br/handle/123456789/11914 Thu, 09 Apr 2026 20:39:34 GMT 2026-04-09T20:39:34Z https://repositorio.ufms.br/handle/123456789/11508 Título: Mineração de um peptídeo com potencial antimicrobiano e anticâncer a partir da sequência do inibidor de tripsina de Inga laurina (ILTI) Abstract: Antimicrobial resistance represents a serious threat to public health, necessitating the urgent development of effective therapeutic alternatives. Among emerging approaches, antimicrobial peptides (AMPs) are particularly promising due to their low propensity to induce resistance, attributed to their direct mechanism of action on the cell membrane, as well as the high efficacy rates observed. This study aimed to develop a peptide with both antimicrobial and anticancer activity, employing computational tools to guide its design and evaluate its activity, toxicity, and structural conformation. The process began with the segmentation and prediction of the amino acid sequence of the ILTI (Inga laurina trypsin inhibitor) protein, using the CAMPR3, NEPTUNE, Sense the Moment, ACPred, AntiCP, Happenn, Ennact, and ToxinPred servers. The top-performing sequence, designated GWA-18, was further refined using the Joker algorithm, which rearranged hydrophobic and hydrophilic residues to generate the bioinspired peptide GWA-17. Following synthesis, the secondary structure of GWA-17 was characterized by circular dichroism, revealing an α-helical conformation in the presence of 50% trifluoroethanol and SDS. Antimicrobial activity was determined using minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays, while cell viability was assessed in BV-2 microglia cells and the tumor cell lines HeLa, B16F10-nex2, SK-mel-28, and A375. GWA-17 demonstrated significant antimicrobial activity, with MIC and MBC values ranging from 4 to 64 μM. Notably, Escherichia coli and Staphylococcus epidermidis exhibited MIC values of 4 μM and 16 μM, respectively. A membrane permeability assay, using the Sytox™ Green fluorescent probe, confirmed that GWA-17 disrupts the bacterial membrane, with noticeable effects within the first 5 minutes of incubation, suggesting a rapid and effective mechanism of action. In cell viability assays, GWA-17 reduced the viability of the tested cancer cell lines, with IC50 values of 21.63 μM for B16F10-nex2, 30.28 μM for HeLa, 51.59 μM for SK-mel-28, and 29.57 μM for A375, inducing apoptotic cell death. Additionally, IC50 values of 31.57 μM for FN1 and 32.72 μM for normal BV-2 cells were observed. Furthermore, GWA-17 exhibited a favorable safety profile, displaying minimal hemolytic activity up to 50 μM, a concentration exceeding the therapeutic range established in previous assays. These findings highlight GWA-17 as a promising bifunctional candidate, exhibiting both antibacterial and anticancer activity, particularly in resistance scenarios, with potential clinical applications in infectious disease control and cancer therapy. Tipo: Dissertação Wed, 01 Jan 2025 00:00:00 GMT https://repositorio.ufms.br/handle/123456789/11508 2025-01-01T00:00:00Z https://repositorio.ufms.br/handle/123456789/11121 Título: PEPaD: um peptídeo multifuncional com atividade antifúngica e anticâncer Abstract: Fungal infections and cancer represent two distinct threats to human health, but in some cases, they are related. Recurrent fungal infections and those caused by other specific microorganisms can induce the mutation of certain cells and consequently lead to the development of various types of cancer. On the other hand, patients undergoing cancer treatment are immunocompromised, which favors infections by microorganisms. Adding this complex link to the growing resistance of fungi and cancer cells to drugs, the development of new active substances or drugs as therapeutic alternatives becomes necessary. Through the broth microdilution technique, it was observed that PEPAD exhibited activity against 5 species of Candida fungi, with MICs ranging between 2.5 - 5 µM, and showed a rapid action time (60 minutes) against Candida tropicalis. It was also found that its likely mechanism of action is through binding with ergosterol present in the plasma membrane. Furthermore, PEPAD showed synergistic action with the antifungal drugs amphotericin B and fluconazole, significantly reducing the required concentrations of both molecules. Inhibition assays of biofilm formation and eradication of mature biofilm were also conducted, and it was found that PEPAD was able to inhibit and eradicate part of the C. tropicalis biofilm. To evaluate the anticancer potential and toxicity of the peptide, cell viability assays were performed using the colorimetric method with MTT. The IC50 found for murine melanoma cells (B16F10-Nex2) was 7.4 µM, whereas for healthy murine macrophage (RAW 264.7) and human fibroblast (FN1) cells, it was not possible to find the IC50 up to the highest tested concentration (16 µM), suggesting high selectivity and low toxicity. After treatment with PEPAD in murine melanoma cells, no morphological alterations were observed in the cell membrane, but changes in nuclear chromatin were noted, suggestive of apoptosis. Through a scratch assay performed on a cell layer, it was observed that PEPAD delayed the cell migration of melanoma, indicating a potential anti-metastatic effect. To evaluate mitochondrial potential and nuclear morphology, the fluorescent probes MitoTracker Deep Red and NucBlue were used. It was observed that the cell nuclei remained intact; however, mitochondrial swelling was observed. Caspase activation was analyzed using the CaspACETM FITC-VAD-FMK marker, revealing active caspases through the binding of the fluorescent marker to activated caspases in cells undergoing apoptotic death. Flow cytometry experiments were also conducted to confirm the type of cell death. The results support previous findings, indicating that PEPAD induces cell death by apoptosis. Finally, ELISA assays were conducted, and it was found that PEPAD was able to induce the release of calreticulin and HMGB1, which are molecules known as Damage-Associated Molecular Patterns (DAMPs), a typical effect of immunogenic cell death. The results suggest the high potential of PEPAD to be used as a prototype in the development of new antifungal and anticancer agents, given its selective characteristics against microorganisms and cancer cells. Tipo: Tese Mon, 01 Jan 2024 00:00:00 GMT https://repositorio.ufms.br/handle/123456789/11121 2024-01-01T00:00:00Z https://repositorio.ufms.br/handle/123456789/9196 Título: Utilização de fungos filamentosos na biorremediação da 3,4-dicloroanilina e atrazina Abstract: Aromatic amines (AA) are one of the classes of compounds most used by industry, in the manufacture of various products, and are among the pollutants most found in both soil and water. The majority of pollutants in this class are derived from dyes and pesticides such as atrazine (ATZ) and 3,4-dichloroaniline (3,4-DCA). One of the alternatives for decontaminating soil and water containing toxic AA is bioremediation, a technique that uses plants, microorganisms or their enzymes to treat polluted sites and recover their original condition. There are few studies using filamentous fungi in AA bioremediation processes, and N-acetylation can be used as a tool, which is an enzymatic process catalyzed by the enzyme arylamine N-acetyltransferase (NAT) found in fungi and other organisms. Thus, the present project aimed to evaluate the capacity of filamentous fungi from the Pantanal and Cerrado mycobiota in the bioremediation of ATZ and 3,4-DCA. A selection was made from 13 strains of filamentous fungi from the genera Aspergillus, Chaetomium, Cladosporium, Mucor and Trichoderma to evaluate the ability to biotransform AA, the 4 strains selected for 3,4-DCA and 9 strains for ATZ. They were subjected to biotransformation tests, where the strains tolerated a concentration of 500 μM for 3,4-DCA, promoting biotransformation into N-acetylated 3,4-DCA (3,4-DCA-Ac) and reduced phytotoxicity in experiments with Lactuca sativa seeds. The conversion of AA into N-acetylated products (AA-Ac) also proved to be efficient in reducing cytotoxic and genotoxic effects in tests carried out with the human keratinocyte cell line (HaCaT). In biotransformation with ATZ, the strains tolerated a concentration of 232 mM, except for A. terreus 31, which had a maximum tolerated concentration of 1 mM. When comparing morphological aspects of the liquid culture medium, mycelial growth and pHs, the strains did not show morphological differences in biomass between control and samples with ATZ, they showed total consumption of ATZ through CCD monitoring and the medium was acidified, changing the pH. In bioremediation tests in plates and soil, the strains reduced the phytotoxic effects of ATZ through the absorption mechanism, confirmed by the higher percentage of ATZ extracted from the mycelial mass after 5 days of experimentation in analysis by HPLC-HRMS ESI/TOF, with the exception of Trichoderma harzianum 27, where the highest percentage was found in the extracellular environment. However, among the 9 strains tested to bioremediate ATZ, the fungi Aspergillus niger and Mucor sp. showed the best results in reducing the phytotoxicity and absorption capacity of ATZ, proving to be promising candidates for moving forward with ATZ bioremediation studies. Tipo: Tese Mon, 01 Jan 2024 00:00:00 GMT https://repositorio.ufms.br/handle/123456789/9196 2024-01-01T00:00:00Z https://repositorio.ufms.br/handle/123456789/9161 Título: Desenvolvimento de gel de carboidrato contendo beterraba e cajá: efeitos em parâmetros cardiorrespiratórios e na percepção subjetiva ao esforço de ciclistas Abstract: When exercising, as energy stores are reduced, there is less aerobic resistance, causing fatigue and hypoglycemia, especially in long training sessions, such as cycling, requiring the need for carbohydrate supplementation. In this sense, the use of a carbohydrate gel supplement becomes interesting due to its practicality in consuming during training and competitions. Aiming for a more natural composition, two constituents are of interest, beetroot and cajá. Because it is rich in nitrate, and under appropriate pH conditions, it gives a natural purplish-red color, and cajá, a fruit found in the Cerrado, rich in antioxidants, with an ideal pH for the stability of deficiency pigments, has there is synergy between the ingredients, favoring the achievement of a “clean label” product. The objective of this study was to develop a carbohydrate gel supplement contained in cajá and carry out nutritional intervention with cyclists, evaluating cardiorespiratory parameters and subjective perception of exertion. The study is characterized as a planned clinical trial, of the crossover type, double-blind, with non-probabilistic sampling, for convenience, approved by the ethics committee of the Federal University of Mato Grosso do Sul. Gels containing cajá pulp were prepared, with levels of carbohydrate of 65% (F1 and F2) and 70% (F3 and F4), varying the content of cajá pulp and pulp between 1.7 and 2.5%, maltodextrin between 50 and 60% and sugar between 10 and 15%. Physicochemical and microbiological characterization was carried out. Sensory characterization occurred in three stages, in the 1st stage F1, F2, F3 and F4 were compared; in the 2nd stage, F1 and F2 were compared (more accepted in the 1st stage) and in the 3rd, F1 and the standard gel were compared. In the nutritional intervention, the F1 gel (65% carbohydrates) and the standard gel were applied, in two controlled 50-minute training sessions, offering 30 g of gel every 20 minutes of training. The training sessions took place a week apart. Heart rate and blood pressure assessments were carried out before and after training, as well as assessment of subjective perception of exertion (RPE). The collected data were analyzed considering a 5% level of significance. The standard gel presented higher moisture, ash, protein and lower carbohydrate content than formulations F2, F3 and F4, not differing from F1. The results of the sensory analyzes showed that the formulated gels were well accepted, without showing any statistical difference (p>0.05) between them. The F1 gel (69.13±2.68 g of carbohydrates) was used in the intervention as it had carbohydrate levels closer to the standard gel (66.68±0.70 g). Microbiological analyzes of Salmonella sp., Enterobacteria, molds and yeasts were absent or lower than the reference standard. Ten male cyclists participated in the study, with a mean age of 39,50 ± 11,42 years. There was a statistically significant difference between HR before and after training 1 (p=0.0001) and training 2 (p=0.002). Only in training 2, SBP (p=0.012) and DBP (p=0.048) decreased. There was no statistically significant difference between having consumed gel A (standard) or B (developed gel) for the RPE, HR and BP values before and after training 1 and training 2. For maximum and average HR, there was also no difference between having consumed gel A or B, for both training sessions. It is concluded that both gels were similar in terms of their effects on cardiovascular parameters, demonstrating that the addition of beetroot and cajá, although in low quantities, were efficient when compared to the standard gel, in addition to contributing to obtaining a more natural supplement and innovative, with fruit from the Cerrado, positively impacting the regional bioeconomy. Key words: Cerrado Fruits; Nutritional supplementation; Athletes; Sports nutrition. Tipo: Dissertação Mon, 01 Jan 2024 00:00:00 GMT https://repositorio.ufms.br/handle/123456789/9161 2024-01-01T00:00:00Z