RESUMO

Skin wound healing is coordinated by a delicate balance between proinflammatory and anti-inflammatory responses, which can be affected by opportunistic pathogens and metabolic or vascular diseases. Several antimicrobial peptides (AMPs) possess immunomodulatory properties, suggesting their potential to support skin wound healing. Here, we evaluated the proregenerative activity of three recently described AMPs (Clavanin A, Clavanin-MO, and Mastoparan-MO). Human primary dermal fibroblasts (hFibs) were used to determine peptide toxicity and their capacity to induce cell proliferation and migration. Furthermore, mRNA analysis was used to investigate the modulation of genes associated with skin regeneration. Subsequently, the regenerative potential of the peptides was further confirmed using an ex vivo organotypic model of human skin (hOSEC)-based lesion. Our results indicate that the three molecules evaluated in this study have regenerative potential at nontoxic doses (i.e., 200 µM for Clavanin-A and Clavanin-MO, and 6.25 µM for Mastoparan-MO). At these concentrations, all peptides promoted the proliferation and migration of hFibs during in vitro assays. Such processes were accompanied by gene expression signatures related to skin regenerative processes, including significantly higher KI67, HAS2 and CXCR4 mRNA levels induced by Clavanin A and Mastoparan-MO. Such findings translated into significantly accelerated wound healing promoted by both Clavanin A and Mastoparan-MO in hOSEC-based lesions. Overall, the data demonstrate the proregenerative properties of these peptides using human experimental skin models, with Mastoparan-MO and Clavanin A showing much greater potential for inducing wound healing compared to Clavanin-MO.

Assuntos

RESUMO

Introduction: The identification of chemical compounds that interfere with SARS-CoV-2 replication continues to be a priority in several academic and pharmaceutical laboratories. Computational tools and approaches have the power to integrate, process and analyze multiple data in a short time. However, these initiatives may yield unrealistic results if the applied models are not inferred from reliable data and the resulting predictions are not confirmed by experimental evidence. Methods: We undertook a drug discovery campaign against the essential major protease (MPro) from SARS-CoV-2, which relied on an in silico search strategy -performed in a large and diverse chemolibrary- complemented by experimental validation. The computational method comprises a recently reported ligand-based approach developed upon refinement/learning cycles, and structure-based approximations. Search models were applied to both retrospective (in silico) and prospective (experimentally confirmed) screening. Results: The first generation of ligand-based models were fed by data, which to a great extent, had not been published in peer-reviewed articles. The first screening campaign performed with 188 compounds (46 in silico hits and 100 analogues, and 40 unrelated compounds: flavonols and pyrazoles) yielded three hits against MPro (IC50 ≤ 25 µM): two analogues of in silico hits (one glycoside and one benzo-thiazol) and one flavonol. A second generation of ligand-based models was developed based on this negative information and newly published peer-reviewed data for MPro inhibitors. This led to 43 new hit candidates belonging to different chemical families. From 45 compounds (28 in silico hits and 17 related analogues) tested in the second screening campaign, eight inhibited MPro with IC50 = 0.12-20 µM and five of them also impaired the proliferation of SARS-CoV-2 in Vero cells (EC50 7-45 µM). Discussion: Our study provides an example of a virtuous loop between computational and experimental approaches applied to target-focused drug discovery against a major and global pathogen, reaffirming the well-known "garbage in, garbage out" machine learning principle.

RESUMO

In skin lesions, the development of microbial infection affects the healing process, increasing morbidity and mortality rates in patients with severe burns, diabetic foot, and other types of skin injuries. Synoeca-MP is an antimicrobial peptide (AMP) that exhibits activity against several bacteria of clinical importance, but its cytotoxicity can represent a problem for its positioning as an effective antimicrobial compound. In contrast, the immunomodulatory peptide IDR-1018 presents low toxicity and a wide regenerative potential due to its ability to reduce apoptotic mRNA expression and promote skin cell proliferation. In the present study, we used human skin cells and a 3D skin equivalent models to analyze the potential of the IDR-1018 peptide to attenuate the cytotoxicity of synoeca-MP, as well as the influence of synoeca-MP/IDR-1018 combination on cell proliferation, regenerative processes, and wound repair. We found that the addition of IDR-1018 significantly improved the biological properties of synoeca-MP on skin cells without modifying its antibacterial activity against S. aureus. Likewise, in both melanocytes and keratinocytes, the treatment with synoeca-MP/IDR-1018 combination induces cell proliferation and migration, while in a 3D human skin equivalent model, it can accelerate wound reepithelization. Furthermore, treatment with this peptide combination generates an up-regulation in the expression of pro-regenerative genes in both monolayer cell cultures and in 3D skin equivalents. This data suggests that the synoeca-MP/IDR-1018 combination possesses a good profile of antimicrobial and pro-regenerative activity, opening the door to the development of new strategies for the treatment of skin lesions.

Assuntos

RESUMO

The invasive nature of Toxoplasma gondii is closely related to the properties of its cytoskeleton, which is constituted by a group of diverse structural and dynamic components that play key roles during the infection. Even if there have been numerous reports about the composition and function of the Toxoplasma cytoskeleton, the ultrastructural organization of some of these components has not yet been fully characterized. This study used a detergent extraction process and several electron microscopy contrast methods that allowed the successful isolation of the cytoskeleton of Toxoplasma tachyzoites. This process allowed for the conservation of the structures known to date and several new structures that had not been characterized at the ultrastructural level. For the first time, characterization was achieved for a group of nanofibers that allow the association between the polar apical ring and the conoid as well as the ultrastructural characterization of the apical cap of the parasite. The ultrastructure and precise location of the peripheral rings were also found, and the annular components of the basal complex were characterized. Finally, through immunoelectron microscopy, the exact spatial location of the subpellicular network inside the internal membrane system that forms the pellicle was found. The findings regarding these new structures contribute to the knowledge concerning the biology of the Toxoplasma gondii cytoskeleton. They also provide new opportunities in the search for therapeutic strategies aimed at these components with the purpose of inhibiting invasion and thus parasitism.

Assuntos

RESUMO

Glyphosate [N-(phosphonomethyl)glycine] is the active ingredient in widely used broad-spectrum herbicides. Even though the toxicity mechanism of this herbicide in vertebrates is poorly understood, evidence suggests that glyphosate is an endocrine disruptor capable of producing morphological anomalies as well as cardiotoxic and neurotoxic effects. We used the zebrafish model to assess the effects of early life glyphosate exposure on the development of cartilage and bone tissues and organismal responses. We found functional alterations, including a reduction in the cardiac rate, significant changes in the spontaneous tail movement pattern, and defects in craniofacial development. These effects were concomitant with alterations in the level of the estrogen receptor alpha osteopontin and bone sialoprotein. We also found that embryos exposed to glyphosate presented spine deformities as adults. These developmental alterations are likely induced by changes in protein levels related to bone and cartilage formation.

Assuntos

RESUMO

The Mapocho River's upper basin (Chilean Central Andes) was studied as a proxy of a high-mountainous hydrothermally-altered (HMHA) system comprised by three sub-basins developed over very different rocks and submitted to different anthropic pressure: 1) a natural acid rock drainage (i.e., Yerba Loca), 2) a creek with mining activity in its headwaters and hydrochemically classified as non-affected by acid mine drainage (i.e., San Francisco), and 3) a low metal concentration creek (i.e., Molina). In general terms, the geochemical composition of the clastic sediments was consistent with the geochemistry inferred from the mineralogical study. However, sediments with a smaller grain size showed higher concentrations than the bigger grain size counterparts for elements such as Fe, S, Cu and As. This behavior was particularly evident in the Yerba Loca basin and it was attributed to the seasonal appearance of Fe- and Al-rich precipitates as constituents of the finer sediments. Different methodologies for the calculation of geochemical backgrounds (Tukey's inner fence, TIF; Median + 2*Median Absolute Deviation, MAD; and 95th percentile) were tested. Results suggest that the 95th percentile-method was the most appropriate for this type of mountainous systems. Using the selected methodology, three different geochemical backgrounds were calculated: 1) Yerba Loca basin, 2) Molina basin, and 3) Mapocho Upper basin. When the generated background levels were compared with the Consensus-Based (CB) Sediment Quality Guidelines; Fe, Mn, Zn, Pb, Cu, Cr, Ni and As showed background values that were consistently higher than the values set by the CB Threshold Effect Concentration and, even higher than the CB Probable Effect Concentration for Fe (MUBBackground: 6.78 wt% vs CB PEC: 4.00 wt%; and Cu (MUBBacground: 3387 mg kg-1 vs CB PEC: 149 mg kg-1). The present study clearly states the paramount importance of having a solid geochemical background before any attempt of a sediment risk assessment is made at HMHA regions.

RESUMO

The Excretory/Secretory (ES) proteins of parasites are involved in invasion and colonization of their hosts. In addition, since ES proteins circulate in the extracellular space, they can be more accessible to drugs than other proteins, which makes ES proteins optimal targets for the development of new and better pharmacological strategies. Monogeneans are a group of parasitic Platyhelminthes that includes some pathogenic species problematic for finfish aquaculture. In the present study, 8297 putative ES proteins from four monogenean species which genomic resources are publicly available were identified and functionally annotated by bioinformatic tools. Additionally, for comparative purposes, ES proteins in other parasitic and free-living platyhelminths were identified. Based on data from the monogenean Gyrodactylus salaris, 15 ES proteins are considered potential drug targets. One of them showed homology to 10 cathepsins with known 3D structure. A docking molecular analysis uncovered that the anthelmintic emodepside shows good affinity to these cathepsins suggesting that emodepside can be experimentally tested as a monogenean's cathepsin inhibitor.

Assuntos

RESUMO

Glyphosate, the most commonly used pesticide worldwide, blocks aromatic amino acid biosynthetic pathways and inhibits growth in plants. Although the specific mode of action of glyphosate in animals remains unclear, adverse effects during embryonic development have been reported, including epiboly delays, morphological alterations, and changes in central nervous system development and cardiogenesis. In this study, we suggest a possible toxicity mechanism for this herbicide related to changes in microtubule stability, which could alter the distribution and dynamics of cytoskeleton components. Using zebrafish embryos to evaluate in vivo effects of glyphosate exposure (5, 10, and 50 µg/ml), we found significant reductions in the levels of acetylated α-tubulin (50 µg/ml) and in the polymeric tubulin percentage in zebrafish embryos that had been exposed to 10 and 50 µg/ml glyphosate, without any changes in either the expression patterns of α-tubulin or the stability of actin filaments. These results indicate that high concentrations of glyphosate were associated with reduced levels of acetylated α-tubulin and altered microtubule stability, which may explain some of the neurotoxic and cardiotoxic effects that have been attributed to this herbicide.

RESUMO

OBJECTIVE: To analyze the association between newborn and maternal characteristics and the risk for cesarean section (CS) due to cephalopelvic disproportion (CPD) and non-CPD causes compared to vaginal deliveries (VD) in a sample of infants and mothers from Merida, Yucatan, Mexico. METHODS: The final sample consisted of 3453 single, live, and term infants born between January 2016 and May 2017 at the Maternal-Infant Hospital in Merida and their mothers (aged ≥19 years). The mode of delivery was established as the dependent variables: (a) VD, (b) CS due to CPD, and (c) non-CPD CS. Independent variables were maternal height and weight, the number of previous VD, newborn weight, and neonatal birthweight (BW) index/maternal height index. A multinomial regression model was used to analyze the association between newborn and maternal characteristics and outcome variable. RESULTS: By mode of delivery, 2124 (62%) births corresponded to VD, 1042 (30%) to non-CPDCS, and 287 (8%) to CS due to CPD. Mothers who had CS due to CPD weighed more at the end of their pregnancy and were shorter. Maternal age and weight increased the risk for having CS due to CPD compared to VD and maternal height, and the number of previous VD reduces the risk for experiencing CS due to CPD compared to vaginal births. The relative risk ratio for higher neonatal BW/maternal height index was significant for CS due to CPD and non-CPD CS. CONCLUSION: According to our results from a public hospital in Merida, Mexico, CPD is a result of the interrelation of maternal and fetal size, rather than an independent result of maternal height or BW.

Assuntos

RESUMO

An integral bioprocess to produce lactic acid (LA) from banana peel (BP) was studied. Oxidases produced by Trametes versicolor and hydrolases produced by Aspergillus flavipes and Aspergillus niger saccharified BP at optimal conditions: 230 rpm, 66 g/L BP, and 73.5% (v/v) of enzymatic crude extract (using equal quantities of the enzymatic extracts). At bioreactor scale (1 L), the joint action of oxidases and hydrolases released 18 g/L of reducing sugars (RS) after 24 h (60% corresponded to glucose), consuming the BP polysaccharides. Lactobacillus delbrueckii fermented the released RS, producing 10 g/L of LA; while in the sequential fermentation (inoculating L. delbrueckii after saccharification), 28 g/L of LA were produced, observing an apparent decrease in feedback inhibition of hydrolases below 1.5 g/L of RS. This process is susceptible for upscaling to produce high LA concentrations and represents a platform to utilize agroindustrial wastes to obtain value-added products.

Assuntos