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2022 ◽  
Vol 8 (1) ◽  
pp. 57
Author(s):  
Nórida Vélez ◽  
Lucía Monteoliva ◽  
Zilpa-Adriana Sánchez-Quitian ◽  
Ahinara Amador-García ◽  
Rocío García-Rodas ◽  
...  

In fungi, metals are associated with the expression of virulence factors. However, it is unclear whether the uptake of metals affects their pathogenicity. This study aimed to evaluate the effect of iron/copper in modulating pathogenicity and proteomic response in two clinical isolates of C. neoformans with high and low pathogenicity. Methods: In both isolates, the effect of 50 µM iron and 500 µM copper on pathogenicity, capsule induction, and melanin production was evaluated. We then performed a quantitative proteomic analysis of cytoplasmic extracts exposed to that combination. Finally, the effect on pathogenicity by iron and copper was evaluated in eight additional isolates. Results: In both isolates, the combination of iron and copper increased pathogenicity, capsule size, and melanin production. Regarding proteomic data, proteins with increased levels after iron and copper exposure were related to biological processes such as cell stress, vesicular traffic (Ap1, Vps35), cell wall structure (Och1, Ccr4, Gsk3), melanin biosynthesis (Hem15, Mln2), DNA repair (Chk1), protein transport (Mms2), SUMOylation (Uba2), and mitochondrial transport (Atm1). Increased pathogenicity by exposure to metal combination was also confirmed in 90% of the eight isolates. Conclusions: The combination of these metals enhances pathogenicity and increases the abundance of proteins related to the main virulence factors.


Author(s):  
Lian-Tao Han ◽  
Yu-Juan Wu ◽  
Tong-Bao Liu

The ubiquitin-proteasome system (UPS) is the major protein turnover mechanism that plays an important role in regulating various cellular functions. F-box proteins are the key proteins of the UPS, responsible for the specific recognition and ubiquitination of downstream targets. Our previous studies showed that the F-box protein Fbp1 plays an essential role in the virulence of C. neoformans. However, the molecular mechanism of Fbp1 regulating the virulence of C. neoformans is still unclear. In this study, we analyzed the potential Fbp1 substrates using an iTRAQ-based proteomic approach and identified the zinc-binding protein Zbp1 as a substrate of Fbp1. Protein interaction and stability assays showed that Zbp1 interacts with Fbp1 and is a downstream target of Fbp1. Ubiquitination analysis in vivo showed that the ubiquitination of Zbp1 is dependent on Fbp1 in C. neoformans. Subcellular localization analysis revealed that the Zbp1 protein was localized in the nucleus of C. neoformans cells. In addition, both deletion and overexpression of the ZBP1 gene led to the reduced capsule size, while overexpression has a more significant impact on capsule size reduction. Fungal virulence assays showed that although the zbp1Δ mutants are virulent, virulence was significantly attenuated in the ZBP1 overexpression strains. Fungal load assay showed that the fungal burdens recovered from the mouse lungs decreased gradually after infection, while no yeast cells were recovered from the brains and spleens of the mice infected by ZBP1 overexpression strains. Thus, our results revealed a new determinant of fungal virulence involving the post-translational regulation of a zinc-binding protein.


2021 ◽  
Vol 140 (1) ◽  
Author(s):  
Alexander Ziegler ◽  
Abigail Miller ◽  
Nina Nagelmann

AbstractThe finned or dumbo octopods (Octopoda: Cirrata) constitute a cephalopod (Mollusca: Cephalopoda) taxon almost exclusively comprising organisms that inhabit the deep-sea. This renders studying the general biology of adult cirrate specimens a difficult task, but even more so when it comes to gathering knowledge on their early life stages. During his comprehensive research exploits on cephalopod development, the late Sigurd von Boletzky (1942–2020) also sought to shed light on specific adaptations that eggs and embryos of these enigmatic deep-sea octopods might show. Based on his seminal work, the present article sets out to provide additional data on a broad range of early cirrate life stages, including egg capsules without any obvious embryonic stage as well as those containing embryos. These previously unreported specimens obtained from museum collections were analyzed using conventional morphological as well as modern three-dimensional imaging techniques. The present overview includes specimens from four of the five oceans, i.e., the Atlantic, Pacific, Indian, and Southern Ocean. Based on information on cirrate ovum, chorion, and egg capsule size as well as shape gathered from the literature, an attempt is made here for the first time to identify von Boletzky's as well as the specimens introduced here down to at least genus level. The combined data provide novel insights into early life stages of finned octopods, aiming to continue von Boletzky's legacy with regard to developmental research on a still largely enigmatic taxon of extant deep-sea cephalopods.


2021 ◽  
Vol 18 (177) ◽  
Author(s):  
A. Said Ismail ◽  
Gregory R. Goodwin ◽  
J. Rafael Castrejon-Pita ◽  
Alastair J. Noyce ◽  
Helena S. Azevedo

In this paper, we demonstrate that aromatic oil capsules, produced by dripping droplets, can offer a simple, yet effective, testing tool to aid in the diagnosis of various diseases, in which the loss of smell is a key symptom. These include chronic neurological conditions such as Parkinson's and Alzheimer's diseases, and acute respiratory infections such as that caused by COVID-19. The capsules were fabricated by concentrically dripping oil/alginate droplets, from a coaxial nozzle, into an oppositely charged ionic liquid. This fabrication technique enables full control over the capsule size, the shell thickness and the volume of the encapsulated oil. After formation, liquid capsules were left to dry and form a solid crust surrounding the oil. The prototype test consists of placing a standardized number of capsules between adhesive strips that users crush and pull apart to release the smell. In addition to the fabrication method, a simple mathematical model was developed to predict the volume of encapsulated oil within the capsule in terms of the flow rate ratio and the nozzle size. Tensile tests show that capsule strength is inversely proportional to its size owing to an increase in the shell thickness. By increasing the alginate concentration, the load required to rupture the capsule increases, to the point where capsules are too stiff to be broken by a fingertip grip. Results from a preliminary screening test, within a group of patients with Parkinson's disease, found that smells were detectable using a ‘forced choice’ paradigm.


Processes ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 452
Author(s):  
Yiyang Wang ◽  
Panagiotis Dimitrakopoulos

The lateral migration of elastic capsules towards a microchannel centerline plays a major role in industrial and physiological processes. Via our computational investigation, we show that a constriction connecting two straight microchannels facilitates the lateral capsule migration considerably, which is relatively slow in straight channels. Our work reveals that the significant cross-streamline migration inside the constriction is dominated by the strong hydrodynamic forces due to the capsule size. However, in the downstream straight channel, the increased interfacial deformation at higher capillary numbers or a lower viscosity ratio and lower membrane hardness results in increased lateral cross-streamline migration. Thus, our work highlights the different migration mechanisms occurring over curved and straight streamlines.


Author(s):  
Annalise Richmond ◽  
David Schwebel ◽  
Casey Morgan ◽  
Gerard Stijntjes

2021 ◽  
Vol 8 ◽  
Author(s):  
India Middlebrook ◽  
Joe Peacock ◽  
Daniel J. Tinnion ◽  
Nicholas K. Leach ◽  
Nathan P. Hilton ◽  
...  

Introduction: Sodium bicarbonate (NaHCO3) is a well-established nutritional ergogenic aid that is typically ingested as a beverage or consumed in gelatine capsules. While capsules may delay the release of NaHCO3 and reduce gastrointestinal (GI) side effects compared with a beverage, it is currently unclear whether the capsule size may influence acid–base responses and GI symptoms following supplementation.Aim: This study aims to determine the effects of NaHCO3 supplementation, administered in capsules of different sizes, on acid–base responses, GI symptoms, and palatability.Methods: Ten healthy male subjects (mean ± SD: age 20 ± 2 years; height 1.80 ± 0.09 m; weight 78.0 ± 11.9 kg) underwent three testing sessions whereby 0.3 g NaHCO3/kg of body mass was consumed in either small (size 3), medium (size 0), or large (size 000) capsules. Capillary blood samples were procured pre-ingestion and every 10 min post-ingestion for 180 min. Blood samples were analyzed using a radiometer (Radiometer ABL800, Denmark) to determine blood bicarbonate concentration ([HCO3-]) and potential hydrogen (pH). GI symptoms were measured using a questionnaire at the same timepoints, whereas palatability was recorded pre-consumption.Results: Capsule size had a significant effect on lag time (the time [HCO3-] changed, Tlag) and the timing of peak blood [HCO3-] (Tmax). Bicarbonate Tlag was significantly higher in the large-sized (28 ± 4 min) compared with the small-sized (13 ± 2 min) capsules (P = 0.009). Similarly, Tmax was significantly lower in the small capsule (94 ± 24 min) compared with both the medium-sized (141 ± 27 min; P < 0.001) and the large-sized (121 ± 29 min; P < 0.001) capsules. The GI symptom scores were similar for small-sized (3 ± 3 AU), medium-sized (5 ± 3 AU), and large-sized (3 ± 3 AU) capsules, with no significant difference between symptom scores (F = 1.3, P = 0.310). Similarly, capsule size had no effect on palatability (F = 0.8, P = 0.409), with similar scores between different capsule sizes.Conclusion: Small capsule sizes led to quicker Tlag and Tmax of blood [HCO3-] concentration compared to medium and large capsules, suggesting that individuals could supplement NaHCO3 in smaller capsules if they aim to increase extracellular buffering capacity more quickly.


PLoS Genetics ◽  
2021 ◽  
Vol 17 (1) ◽  
pp. e1009313
Author(s):  
Cullen Roth ◽  
Debra Murray ◽  
Alexandria Scott ◽  
Ci Fu ◽  
Anna F. Averette ◽  
...  

Cryptococcal disease is estimated to affect nearly a quarter of a million people annually. Environmental isolates of Cryptococcus deneoformans, which make up 15 to 30% of clinical infections in temperate climates such as Europe, vary in their pathogenicity, ranging from benign to hyper-virulent. Key traits that contribute to virulence, such as the production of the pigment melanin, an extracellular polysaccharide capsule, and the ability to grow at human body temperature have been identified, yet little is known about the genetic basis of variation in such traits. Here we investigate the genetic basis of melanization, capsule size, thermal tolerance, oxidative stress resistance, and antifungal drug sensitivity using quantitative trait locus (QTL) mapping in progeny derived from a cross between two divergent C. deneoformans strains. Using a “function-valued” QTL analysis framework that exploits both time-series information and growth differences across multiple environments, we identified QTL for each of these virulence traits and drug susceptibility. For three QTL we identified the underlying genes and nucleotide differences that govern variation in virulence traits. One of these genes, RIC8, which encodes a regulator of cAMP-PKA signaling, contributes to variation in four virulence traits: melanization, capsule size, thermal tolerance, and resistance to oxidative stress. Two major effect QTL for amphotericin B resistance map to the genes SSK1 and SSK2, which encode key components of the HOG pathway, a fungal-specific signal transduction network that orchestrates cellular responses to osmotic and other stresses. We also discovered complex epistatic interactions within and between genes in the HOG and cAMP-PKA pathways that regulate antifungal drug resistance and resistance to oxidative stress. Our findings advance the understanding of virulence traits among diverse lineages of Cryptococcus, and highlight the role of genetic variation in key stress-responsive signaling pathways as a major contributor to phenotypic variation.


2021 ◽  
Vol 22 (2) ◽  
pp. 584
Author(s):  
Amani Zoabi ◽  
Katherine Margulis

(1) Background: Chiral nanoparticular systems have recently emerged as a compelling platform for investigating stereospecific behavior at the nanoscopic level. We describe chiroselective supramolecular interactions that occur between DNA oligonucleotides and chiral polyurea nanocapsules. (2) Methods: We employ interfacial polyaddition reactions between toluene 2,4-diisocyanate and lysine enantiomers that occur in volatile oil-in-water nanoemulsions to synthesize hollow, solvent-free capsules with average sizes of approximately 300 nm and neutral surface potential. (3) Results: The resultant nanocapsules exhibit chiroptical activity and interact differentially with single stranded DNA oligonucleotides despite the lack of surface charge and, thus, the absence of significant electrostatic interactions. Preferential binding of DNA on d-polyurea nanocapsules compared to their l-counterparts is demonstrated by a fourfold increase in capsule size, a 50% higher rise in the absolute value of negative zeta potential (ζ-potential), and a three times lower free DNA concentration after equilibration with the excess of DNA. (4) Conclusions: We infer that the chirality of the novel polymeric nanocapsules affects their supramolecular interactions with DNA, possibly through modification of the surface morphology. These interactions can be exploited when developing carriers for gene therapy and theranostics. The resultant constructs are expected to be highly biocompatible due to their neutral potential and biodegradability of polyurea shells.


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