Biology subjects

da Silva, M.

Publications and source records attributed to da Silva, M..

5 recordsLinked to original sources

bioRxiv2026

Mapping virulence-associated protein interaction networks reveals regulators of thermotolerance in Cryptococcus neoformans

Protein-protein interactions (PPIs) influence critical biological processes in pathogenic microorganisms, such as the human fungal pathogen, Cryptococcus neoformans. Fungal thermotolerance and stress response pathways are key virulence determinants that directly impact pathogen adaptation and survival and the infection process. To establish a comprehensive baseline of PPIs in C. neoformans and explore these interactions to infer functional roles for uncharacterized proteins, we applied size exclusion chromatography coupled with mass spectrometry to the secreted and cellular proteomes of the fungi. As a result, 216 and 1699 unique proteins were identified across 24 secretome and proteome fractions, respectively. The predicted secretome networks included expected proteins associated with vesicles and virulence, indicating a role in extracellular defense. Whereas the cryptococcal proteome highlighted interactions among proteins with defined roles in fungal virulence for protein stability and thermotolerance, including two previously uncharacterized proteins, CNAG_00287 and CNAG_05199, putatively involved in complex formation with heat-shock proteins (HSP). Based on sequence and structure homology, we propose that CNAG_00287 is a tetratricopeptide repeat-containing co-chaperone that modulates Hsp 70 activity and CNAG_05199 functions as a Hsp70. We validated the thermotolerance role of CNAG_00287 in heat-related stress, as its absence significantly impaired fungal growth in nutrient-limited media at 37 {degrees}C. Together, this work resolves virulence-associated PPIs within C. neoformans and reveals new molecular regulators of thermotolerance that underpin fungal pathogenicity.

microbiology↗

bioRxiv2025

Differential patterns of cortical expansion in fetal and preterm brain development

The human cerebral cortex undergoes a complex developmental process during gestation, characterised by rapid cortical expansion and gyrification. This study investigates in vivo cortical growth trajectories using longitudinal MRI data from fetal and preterm cohorts. We employed anatomically constrained multimodal surface matching (aMSM) to quantify cortical surface area expansion and compare in-utero versus ex-utero cortical growth using cortical surface data from 22 to 44 weeks post-menstrual age (PMA), acquired as part of the Developing Human Connectome Project (dHCP). Our findings revealed distinct regional and temporal growth patterns during the 2nd and 3rdrd trimesters of healthy fetal cortical expansion. Ex utero brain development following preterm birth was shown to follow a modified trajectory compared to normal gestation, with potential implications for cortical organisation. Our methodology, combining biomechanically constrained surface registration with high quality fetal and neonatal imaging, provides a powerful framework for understanding early cortical development and deviations associated with preterm birth.

neuroscience↗

bioRxiv2022

Multi-lingual multi-platform investigations of online trade in jaguar parts

We conducted research to understand online trade in jaguar parts and develop tools of utility for jaguars and other species. Our research took place to identify potential trade across 31 online platforms in Spanish, Portuguese, English, Dutch, French, Chinese, and Vietnamese. We identified 230 posts from between 2009 and 2019. We screened the images of animal parts shown in search results to verify if from jaguar; 71 posts on 12 different platforms in four languages were accompanied by images identified as definitely jaguar, including a total of 125 jaguar parts (50.7% posts in Spanish, 25.4% Portuguese, 22.5% Chinese and 1.4% French). Search effort varied among languages due to staff availability. Standardizing for effort across languages by dividing number of posts advertising jaguars by search time and number individual searches completed via term/platform combinations, the adjusted rankings of posts were: Portuguese #1, Chinese 2 (time) & 3 (searches), Spanish 3 & 4; French 5 & 4; English 5 & 2, and Dutch 6. Teeth were most common; 156 posts offered at least 367 apparent teeth. From these, 95 teeth were assessed as definitely jaguar; 71 jaguar teeth could be linked to a location, with the majority of the 71 offered for sale from Mexico, China, Bolivia, and Brazil (26.8, 25.4, 16.9, and 12.7% respectively). Ranking of number of teeth was Mexico (19), China (18), Bolivia (12), Brazil (9), Peru/Ecuador (most accurate probable location) (8), Venezuela (3), Guadeloupe (1), and Uruguay (1). The second most traded item, skins and derivative items were only identified from Latin America: Brazil (7), was followed by Peru (6), Bolivia (3), Mexico (2 and 1 skin piece), and Nicaragua and Venezuela (1 each). Whether by number of posts or pieces, the ranking of parts was teeth, skins/pieces of skins, heads, and bodies. Our research presents a snapshot of online jaguar trade and methods that may have utility for many species now traded online. Our research took place within a longer-term project to assist law enforcement in host countries to better identify potential illegal trade online, with research findings informing hubs in Latin America for building such capacity.

ecology↗

bioRxiv2019

Drug reformulation for a neglected disease. The NANOHAT project to develop a safer more effective sleeping sickness drug.

Human African trypanosomiasis (HAT or sleeping sickness) is caused by the parasite Trypanosoma brucei sspp. The disease has two stages, a haemolymphatic stage after the bite of an infected tsetse fly, followed by a central nervous system stage where the parasite penetrates the brain, causing death if untreated. Treatment is stage-specific, due to the blood-brain barrier, with less toxic drugs such as pentamidine used to treat stage 1. The objective of our research programme was to develop an intravenous formulation of pentamidine which increases CNS exposure by some 10-100 fold, leading to efficacy against a model of stage 2 HAT. This target candidate profile is in line with drugs for neglected diseases inititative recommendations. To do this, we evaluated the physicochemical and structural characteristics of formulations of pentamidine with Pluronic micelles (triblock-copolymers of polyethylene-oxide and polypropylene oxide), selected candidates for efficacy and toxicity evaluation in vitro, quantified pentamidine CNS delivery of a sub-set of formulations in vitro and in vivo, and progressed one pentamidine-Pluronic formulation for further evaluation using an in vivo single dose brain penetration study. Screening pentamidine against 40 CNS targets did not reveal any major neurotoxicity concerns, however, pentamidine had a high affinity for the imidazoline2 receptor. The reduction in insulin secretion in MIN6 {beta}-cells by pentamidine maybe secondary to pentamidine-mediated activation of {beta}-cell imidazoline receptors and impairment of cell viability. Pluronic F68 (0.01%w/v)-pentamidine formulation had a similar inhibitory effect on insulin secretion as pentamidine alone and an additive trypanocidal effect in vitro. However, all Pluronics tested (P85, P105 and F68) did not significantly enhance brain exposure of pentamidine. These results are relevant to further developing block-copolymers as nanocarriers, improving BBB drug penetration and understanding the side effects of pentamidine.

pharmacology and toxicology↗

bioRxiv2018

Transcription activity contributes to the activation of non-constitutive origins to maintain the robustness of S phase duration in African trypanosomes

The cosynthesis of DNA and RNA potentially generates conflicts between replication and transcription, which can lead to genomic instability. In trypanosomatids, eukaryotic parasites that perform polycistronic transcription, this phenomenon and its consequences have not yet been investigated. Here, using equations and computational analysis we demonstrated that the number of constitutive origins mapped in the Trypanosoma brucei genome is close to the minimum required to complete replication within S phase duration. However, taking into account the location of these origins in the genome, the replication in due time becomes virtually impossible, making it necessary to activate non-constitutive origins. Moreover, computational and biological assays pointed to transcription being responsible for activating non-constitutive origins. Together, our results suggest that transcription action through conflicts with replication contributes to the firing of non-constitutive origins, maintaining the robustness of S phase duration. The usage of this entire pool of origins seems to be of paramount importance for the survival of this parasite that infects million people around the world since it contributes to the maintenance of the replication of its DNA.

molecular biology↗