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Biology subjects

Ferreira, R. S.

Publications and source records attributed to Ferreira, R. S..

7 recordsLinked to original sources

NoiTox: A Machine Learning Strategy for Peptide Toxicity Prediction Reveals Label Noise in Benchmark Datasets

Motivation: Peptide toxicity remains a major bottleneck in drug development, as small sequence variations can drastically alter cytotoxic profiles. Experimental assessment is costly and time-consuming, motivating computational predictors. Existing approaches often rely on single black-box architectures, limiting interpretability and obscuring the contribution of individual sequence features. Furthermore, benchmark datasets frequently define the negative class through annotation filters rather than direct experimental evidence, introducing structural label noise that can compromises training and evaluation. Results: We present NoiTox, a multi-model strategy for peptide toxicity prediction integrating three complementary architectures, evaluated as four model variants: NoiToxML (Bo/full), an Extremely Randomized Trees classifier with Boruta-selected or complete physicochemical, compositional, and atomic descriptors; NoiToxDP, a deep learning model leveraging ESM-2 protein language model embeddings; and NoiToxGP, a graph neural network that combines ESM-2 residue embeddings, a fully connected residue graph with attention, and the Boruta descriptors as global features. Averaged over five random seeds, the models approached the state-of-the-art predictor ToxiPep on CD-HIT-controlled test sets, and on an independent dataset the interpretable NoiToxML variants and a probability-averaging consensus of the four models exceeded it, showing that physicochemical descriptors are highly informative for toxicity prediction. Cross-referencing negative-labeled sequences against curated databases of hemolytic, cytotoxic, and neurotoxic peptides flagged a small set annotated as toxic elsewhere, consistent with label noise by omission. However, removing the sequences that all models contradicted did not improve, and for some models reduced, ranking performance, indicating that such sequences carry useful information and are better treated as candidates for curation than discarded automatically. Availability and implementation: https://github.com/goalmeida05/NoiTox.

bioinformatics↗

Discovery of Semicarbazone and Thiosemicarbazone Analogs as Competitive SARS-CoV-2 Virus Main Protease (Mpro) Inhibitors

Despite the development of vaccines and antivirals, coronavirus disease 2019 (COVID-19) continues to affect populations worldwide. Given the high mutation rate of the SARS-CoV-2 virus and reports of drug resistance, there is a continued need for new therapeutic options. SARS-CoV-2 main protease (Mpro) is essential for viral replication and is a conserved target among coronaviruses. Most known Mpro inhibitors target the active site, although allosteric sites have already been identified. In this study, we conducted a virtual screening of 2,060 compounds targeting an allosteric site of SARS-CoV-2 Mpro. From this screen, 41 computational hits and analogs were selected and evaluated using biochemical assays against SARS-CoV-2 Mpro. Among them, compound 25, a semicarbazone, demonstrated a half-maximal inhibitory concentration (IC50) of 99 M. Additionally, two thiosemicarbazone analogs (compounds 50 and 51) inhibited SARS-CoV-2 Mpro with IC50 values of 61 M and 70 M. Biochemical assays suggest that these compounds act as noncovalent competitive inhibitors of SARS-CoV-2 Mpro. Molecular dynamics simulations revealed that compound 25 is unstable at the allosteric site of SARS-CoV-2 Mpro but forms stable and favorable interactions at the active site, supporting its potential as a competitive inhibitor, a finding subsequently confirmed by biochemical assays. Our structure-based computational and biochemical approach identified semicarbazone and thiosemicarbazone scaffolds as promising candidates for the development of reversible SARS-CoV-2 Mpro inhibitors.

biochemistry↗

Metabolic and age-associated epigenetic barriers during direct reprogramming of mouse fibroblasts into induced cardiomyocytes

Heart disease is the leading cause of mortality in developed countries novel regenerative procedures are warranted for improving patients well fare. Direct cardiac conversion (DCC) can create induced cardiomyocytes (iCMs) and besides holding great promise, lacks clinical effectiveness for cardiac regeneration as metabolic and age-associated barriers remains elusive. Here, we identify by histone post-translational analysis that DCC triggers major age-dependent alterations in the epigenetic landscape. Metabolomics revealed decrease abundance of anabolic metabolites related to TCA cycle and glutaminolysis and profound mitochondrial network remodeling, with increased elongation and interconnectivity and reliance in mitochondrial respiration in iCMs. Importantly, adult-derived iCMs present increase accumulation of oxidative stress in the mitochondria and pharmacological activation of mitophagy increase DCC of adult fibroblasts in vitro. Metabolic modulation in vitro and dietary manipulations in vivo improves DCC efficiency and are accompanied by significant alterations in the histone acetylation and methylation landscape and mitochondria homeostasis. Our study provides evidence that metaboloepigenetics as a direct role in cell fate transitions driving direct cardiac conversion into iCMs, highlighting the potential use of metabolic modulation in increasing cardiac regenerative strategies.

cell biology↗

Prioritizing virtual screening with interpretable interaction fingerprints

Machine learning-based drug discovery success depends on molecular representation. Yet traditional molecular fingerprints omit both the protein and pointers back to structural information that would enable better model interpretability. Therefore, we propose LUNA, a Python 3 toolkit that calculates and encodes protein-ligand interactions into new hashed fingerprints inspired by Extended Connectivity Finger-Print (ECFP): EIFP (Extended Interaction FingerPrint), FIFP (Functional Interaction FingerPrint), and Hybrid Interaction FingerPrint (HIFP). LUNA also provides visual strategies to make the fingerprints interpretable. We performed three major experiments exploring the fingerprints use. First, we trained machine learning models to reproduce DOCK3.7 scores using 1 million docked Dopamine D4 complexes. We found that EIFP-4,096 performed (R2 = 0.61) superior to related molecular and interaction fingerprints. Secondly, we used LUNA to support interpretable machine learning models. Finally, we demonstrate that interaction fingerprints can accurately identify similarities across molecular complexes that other fingerprints over-look. Hence, we envision LUNA and its interface fingerprints as promising methods for machine learning-based virtual screening campaigns. LUNA is freely available at https://github.com/keiserlab/LUNA.

bioinformatics↗

SEEKING CANDIDATE MOLECULES AS PROGNOSTIC HEALING MARKERS IN CHRONIC VENOUS ULCERS

Seeking and identifying biomarker molecules in inflammatory exudate of chronic venous ulcers (CVUs) can aid health professionals in the healing prognosis. The therapeutic failure or cure is related to the quantitative expression of determinate proteins. This work aimed to identify the proteins expressed in inflammatory exudates from CVUs and correlate them with reduction or increase in the wound size. For 90 days, 28 participants that received standard treatment for 37 CVUs were monitored. The inflammatory exudates were collected before treatment initiation (T=0) and analyzed via the Label-free Shotgun. After 90 days the wound area was reduced in 25 (67.6%) of them. Mass spectrometry analysis of all the inflammatory exudates showed four proteins differentially expressed and related to favorable or unfavorable evolution of the healing process. Complement C3 and ceruloplasmin were identified in all the lesions analyzed and were expressed differentially in lesions that presented diminished area in the studied period. Apoliprotein A1 and neutrophil defensin-1 presented differential expression in ulcers that either did not diminish or augmented their wound area through 90 days. These results suggest that Complement C3, Ceruloplasmin, Apoliprotein A1 and Neutrophil-defensin-1 proteins are potential candidate molecules for prognostic healing markers in chronic venous ulcers.

biochemistry↗

Improvement of bone regeneration using fibrin biopolymer combined with differentiated stem cells

1.Bone tissue repair remains a challenge on tissue engineering. New approaches are highly expected to regenerate fractures, bone infections, cancers and congenital skeletal abnormalities. Lately, osteoconductive biomaterials have been used with osteoprogenitor cellsas bone substitutes to accelerate bone formation. Fibrin scaffold serves as a provisional platform promoting cell migration and proliferation, angiogenesis, connective tissue formation and growth factors stimulation. When combined with mesenchymal stem cells (MSCs) maintain cell viability that exerts an immunomodulatory effect by modifying inflammatory environment through expression of pro and anti-inflammatory cytokines. We evaluated a unique heterologous fibrin biopolymer as scaffold to MSCs on bone regeneration of rat femurs. A critical-size bone defect was made in the femur and treated with fibrin biopolymer(FBP); FBP + MSC; and FBP + MSC differentiated in bone lineage (MSC-D). Bone repair was analyzed 03, 21 and 42 days later by radiographic, histological and scanning electron microscopy (SEM) imaging. The FBP+MSC-D association was the most effective treatment, since newly formed bone was more abundant and early matured in just 21 days. Our results demonstrate that FBP isolated was able to promote bone repair although cells play a crucial role on the type and quantity of bone tissue formed. We have not observed surgical site infection, inflammatory response, fractures or loss of function related with FBP. Thus, this approach can be safely expanded for clinical trials as an effort to overcome current method limitations and improve overall bone regeneration process.

bioengineering↗

A method for faster purification of serine proteinases from Bothrops alternatus and Bothrops moojeni snake venoms

Snake venoms are important sources of substances with a variety of pharmacological activities. Among the different proteins present in these venoms, snake venom serine proteinases (SVSPs) have important effects on the hemostatic system that influence the hemodynamic properties of blood. Bothrops genus snakes presented their venom richly composed of SVSPs thrombin-like, and the isolation of these enzymes is of great interest. In 1994, the Center for the Study of Venoms and Venomous Animals (CEVAP) - UNESP standardized the fibrin sealant derived from snake venom, replacing the bovine thrombin by gyroxin thrombin-like enzyme from Crotalus durissus terrificus (Rattlesnake) and human plasma fibrinogen by buffaloes cryoprecipitate. Despite chromatographic techniques for the purification of gyroxin be well grounded in the literature, that income is considered low. Thus, in addition to gyroxin, other thrombin-like enzymes could be employed in the composition of the new fibrin sealant after being standardized to the purifying and chromatographic performance and widely evaluated for biological activities. Therefore, it is extremely important that in our lab is deployed, standardized and validated a method for the chromatographic purification of other thrombin-like enzymes such as found in Bothrops snake venoms. Thus a two-step chromatographic procedure was developed to routinely purify serine proteinases from Bothrops alternatus and B. moojeni snakes venoms to provide new enzymes for improving the CEVAPs heterologous fibrin sealant.

biochemistry↗