scholarly journals Comparison of the RNA Content of Extracellular Vesicles Derived from Paracoccidioides brasiliensis and Paracoccidioides lutzii

Cells ◽  
2019 ◽  
Vol 8 (7) ◽  
pp. 765 ◽  
Author(s):  
Roberta Peres da Silva ◽  
Larissa G. V. Longo ◽  
Julia P. C. da Cunha ◽  
Tiago J. P. Sobreira ◽  
Marcio L. Rodrigues ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Protein Modification ◽  
Paracoccidioides Brasiliensis ◽  
Pathogenic Fungi ◽  
In Vitro Translation ◽  
Rna Seq ◽  
Rna Content ◽  
Oxidation Reduction ◽  
Culture Supernatants

Paracoccidioides brasiliensis and P. lutzii cause human paracoccidioidomycosis. We have previously characterized the <200-nt RNA sub-populations contained in fungal extracellular vesicles (EVs) from P. brasiliensis Pb18 and other pathogenic fungi. We have presently used the RNA-seq strategy to compare the <200- and >200-nt RNA fractions contained in EVs isolated from culture supernatants of P. brasiliensis Pb18, Pb3, and P. lutzii Pb01. Shared mRNA sequences were related to protein modification, translation, and DNA metabolism/biogenesis, while those related to transport and oxidation-reduction were exclusive to Pb01. The presence of functional full-length mRNAs was validated by in vitro translation. Among small non-coding (nc)RNA, 15 were common to all samples; small nucleolar (sno)RNAs were enriched in P. brasiliensis EVs, whereas for P. lutzii there were similar proportions of snoRNA, rRNA, and tRNA. Putative exonic sRNAs were highly abundant in Pb18 EVs. We also found sRNA sequences bearing incomplete microRNA structures mapping to exons. RNA-seq data suggest that extracellular fractions containing Pb18 EVs can modulate the transcriptome of murine monocyte-derived dendritic cells in a transwell system. Considering that sRNA classes are involved in transcription/translation modulation, our general results may indicate that differences in virulence among fungal isolates can be related to their distinct EV-RNA content.

scholarly journals The Oviductal Extracellular Vesicles’ RNA Cargo Regulates the Bovine Embryonic Transcriptome

2020 ◽  
Vol 21 (4) ◽  
pp. 1303 ◽  
Author(s):  
Stefan Bauersachs ◽  
Pascal Mermillod ◽  
Carmen Almiñana
Keyword(s):  
Gene Expression ◽  
Extracellular Vesicles ◽  
Sequencing Analysis ◽  
Rna Seq ◽  
High Throughput Analysis ◽  
Positive Effects ◽  
Early Embryos ◽  
And Control ◽  
The Impact

Oviductal extracellular vesicles (oEVs) are emerging as key players in the gamete/embryo–oviduct interactions that contribute to successful pregnancy. Various positive effects of oEVs on gametes and early embryos have been found in vitro. To determine whether these effects are associated with changes of embryonic gene expression, the transcriptomes of embryos supplemented with bovine fresh (FeEVs) or frozen (FoEVs) oEVs during in vitro culture compared to controls without oEVs were analyzed by low-input RNA sequencing. Analysis of RNA-seq data revealed 221 differentially expressed genes (DEGs) between FoEV treatment and control, 67 DEGs for FeEV and FoEV treatments, and minor differences between FeEV treatment and control (28 DEGs). An integrative analysis of mRNAs and miRNAs contained in oEVs obtained in a previous study with embryonic mRNA alterations pointed to direct effects of oEV cargo on embryos (1) by increasing the concentration of delivered transcripts; (2) by translating delivered mRNAs to proteins that regulate embryonic gene expression; and (3) by oEV-derived miRNAs which downregulate embryonic mRNAs or modify gene expression in other ways. Our study provided the first high-throughput analysis of the embryonic transcriptome regulated by oEVs, increasing our knowledge on the impact of oEVs on the embryo and revealing the oEV RNA components that potentially regulate embryonic development.

scholarly journals Extracellular Vesicle-Mediated RNA Release in Histoplasma capsulatum

mSphere ◽  
2019 ◽  
Vol 4 (2) ◽  
Author(s):  
Lysangela R. Alves ◽  
Roberta Peres da Silva ◽  
David A. Sanchez ◽  
Daniel Zamith-Miranda ◽  
Marcio L. Rodrigues ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Stress Responses ◽  
Histoplasma Capsulatum ◽  
Pathogenic Fungi ◽  
Extracellular Vesicle ◽  
Fungal Species ◽  
Cellular Communication ◽  
Content Type ◽  
Rna Molecules ◽  
Rna Content

ABSTRACT Eukaryotic cells, including fungi, release extracellular vesicles (EVs). These lipid bilayered compartments play essential roles in cellular communication and pathogenesis. EV composition is complex and includes proteins, glycans, pigments, and RNA. RNAs with putative roles in pathogenesis have been described in EVs produced by fungi. Here we describe the RNA content in EVs produced by the G186AR and G217B strains of Histoplasma capsulatum, an important human-pathogenic fungal pathogen. A total of 124 mRNAs were identified in both strains. In this set of RNA classes, 93 transcripts were enriched in EVs from the G217B strain, whereas 31 were enriched in EVs produced by the G186AR strain. This result suggests that there are important strain-specific properties in the mRNA composition of fungal EVs. We also identified short fragments (25 to 40 nucleotides in length) that were strain specific, with a greater number identified in EVs produced by the G217B strain. Remarkably, the most highly enriched processes were stress responses and translation. Half of these fragments aligned to the reverse strand of the transcript, suggesting the occurrence of microRNA (miRNA)-like molecules in fungal EVs. We also compared the transcriptome profiles of H. capsulatum with the RNA composition of EVs, and no correlation was observed. Taking the results together, our study provided information about the RNA molecules present in H. capsulatum EVs and about the differences in composition between the strains. In addition, we found no correlation between the most highly expressed transcripts in the cell and their presence in the EVs, reinforcing the idea that the RNAs were directed to the EVs by a regulated mechanism. IMPORTANCE Extracellular vesicles (EVs) play important roles in cellular communication and pathogenesis. The RNA molecules in EVs have been implicated in a variety of processes. EV-associated RNA classes have recently been described in pathogenic fungi; however, only a few reports of studies describing the RNAs in fungal EVs are available. Improved knowledge of EV-associated RNA will contribute to the understanding of their role during infection. In this study, we described the RNA content in EVs produced by two isolates of Histoplasma capsulatum. Our results add this important pathogen to the current short list of fungal species with the ability to use EVs for the extracellular release of RNA.

scholarly journals Fungus-mediated green synthesis of nano-silver using Aspergillus sydowii and its antifungal/antiproliferative activities

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Dongyang Wang ◽  
Baiji Xue ◽  
Lin Wang ◽  
Yidi Zhang ◽  
Lijun Liu ◽  
...  
Keyword(s):  
Color Change ◽  
Cost Effective ◽  
Pathogenic Fungi ◽  
Aspergillus Sydowii ◽  
X Ray Diffraction ◽  
Synthesis Conditions ◽  
Transmission Electron ◽  
Increasing Demand ◽  
Culture Supernatants

AbstractDue to the increasing demand for eco-friendly, cost-effective and safe technologies, biosynthetic metal nanoparticles have attracted worldwide attention. In this study, silver nanoparticles (AgNPs) were extracellularly biosynthesized using the culture supernatants of Aspergillus sydowii. During synthesis, color change was preliminarily judge of the generation of AgNPs, and the UV absorption peak at 420 nm further confirms the production of AgNPs. Transmission electron microscopy and X-ray diffraction were also used to identify the AgNPs. The results shows that AgNPs has crystalline cubic feature and is a polydisperse spherical particle with size between 1 and 24 nm. Three main synthesis factors (temperature, pH and substrate concentration) were optimized, the best synthesis conditions were as follows 50 °C, 8.0 and 1.5 mM. In the biological application of AgNPs, it shows effective antifungal activity against many clinical pathogenic fungi and antiproliferative activity to HeLa cells and MCF-7 cells in vitro. Our research finds a new path to biosynthesis of AgNPs in an eco-friendly manner, and bring opportunity for biomedical applications in clinic.

Characterization and regulation of extracellular vesicles in the lumen of the ovine uterus†

2020 ◽  
Vol 102 (5) ◽  
pp. 1020-1032 ◽  
Author(s):  
Eleanore V O’Neil ◽  
Gregory W Burns ◽  
Christina R Ferreira ◽  
Thomas E Spencer
Keyword(s):  
Extracellular Vesicles ◽  
Cell Types ◽  
Size Exclusion ◽  
Rna Seq ◽  
Interferon Tau ◽  
Uterine Lumen ◽  
Pregnant Sheep ◽  
Exclusion Chromatography ◽  
Ovine Uterus

Abstract Secretions of the endometrium are vital for peri-implantation growth and development of the sheep conceptus. Extracellular vesicles (EVs) are present in the uterine lumen, emanate from both the endometrial epithelia of the uterus and trophectoderm of the conceptus, and hypothesized to mediate communication between those cell types during pregnancy establishment in sheep. Size-exclusion chromatography and nanoparticle tracking analysis determined that total EV number in the uterine lumen increased from days 10 to 14 of the cycle but was lower on days 12 and 14 of pregnancy in sheep. Intrauterine infusions of interferon tau (IFNT) did not affect total EV number in the uterine lumen. Quantitative mass spectrometric analyses defined proteins and lipids in EVs isolated from the uterine lumen of day 14 cyclic and pregnant sheep. In vitro analyses found that EVs decreased ovine trophectoderm cell proliferation and increased IFNT production without effects on gene expression as determined by RNA-seq. Collective results support the idea EVs impact conceptus growth during pregnancy establishment via effects on trophectoderm cell growth.

scholarly journals Extracellular vesicle-mediated RNA release inHistoplasma capsulatum

2019 ◽  
Author(s):  
Lysangela R. Alves ◽  
Roberta Peres da Silva ◽  
David A. Sanchez ◽  
Daniel Zamith-Miranda ◽  
Marcio L. Rodrigues ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Stress Responses ◽  
Histoplasma Capsulatum ◽  
Pathogenic Fungi ◽  
Extracellular Vesicle ◽  
Fungal Species ◽  
Cellular Communication ◽  
Rna Molecules ◽  
Rna Content ◽  
Reverse Strand

AbstractEukaryotic cells, including fungi, release extracellular vesicles (EVs). These lipid bilayered compartments play essential roles in cellular communication and pathogenesis. EV composition is complex and includes proteins, glycans, pigments, and RNA. RNA classes with putative roles in pathogenesis have been described in EVs produced by fungi. Here we describe the RNA content in EVs produced by the G186AR and G217B strains ofHistoplasma capsulatum, an important human fungal pathogen. A total of 124 mRNA were identified in both strains. In this set of RNA classes, 93 transcripts were enriched in EVs from the G217B strain, while 31 enriched in EVs produced by the G186AR strain. This result suggests that there are important strain-specific properties in the mRNA composition of fungal EVs. We also identified short fragments (25-40 long) that were strain-specific, with a greater number of them identified in EVs produced by the G217B strain. Remarkably, the most enriched processes were stress responses and translation. Half of these fragments aligned to the reverse strand of the transcript, suggesting the occurrence of miRNA-like molecules in fungal EVs. We also compared the transcriptome profiles ofH. capsulatumwith the RNA composition of EVs and no correlation was observed. Altogether, our study provided information about the RNA molecules present inH. capsulatumEVs, and the differences in composition between the G186AR and G217B strains. In addition, we showed that the correlation between the most expressed transcripts in the cell and their presence in the EVs, reinforcing the idea that the RNAs were directed to the EVs by a regulated mechanism.ImportanceExtracellular vesicles (EVs) play important roles in cellular communication and pathogenesis. The RNA molecules in EVs have been implicated in a variety of processes. In pathogenic fungi, EV-associated RNA classes have recently been described; however, only a few studies describing the RNA in fungal EVs are available. An improved knowledge on EV-associated RNA will contribute to the understanding of their role during infection. In this study, we described the RNA content in EVs produced by two isolates ofHistoplasma capsulatum. Our results add this important pathogen to the current short list of fungal species with the ability to use EVs for the extracellular release of RNA.

scholarly journals A Transcriptome Profile Reveals The Regulatory Mechanism of Verticillium Dahliae Against Bacillus

2021 ◽  
Author(s):  
Wenhui Tian ◽  
Zhenrui Cheng ◽  
Junxia Wang ◽  
Fengfeng Cheng ◽  
Luping Li ◽  
...  
Keyword(s):  
Verticillium Dahliae ◽  
Regulatory Mechanism ◽  
Pathogenic Fungi ◽  
Oxygen Species ◽  
Rna Seq ◽  
Transcriptome Profile ◽  
Biocontrol Bacteria ◽  
Expression Of Genes ◽  
Prevention And Treatment

Abstract Background: Verticillium dahliae, the causal agent of Verticillium wilt, is notoriously invasive in many crops and has been involved in numerous epidemics worldwide. Bacillus species, as representatives of biocontrol bacteria, produce a variety of lipopeptides (LPs), which are useful as biofungicides to many pathogenic fungi, including Verticillium dahliae. This study will explore the mechanism of resistance of V. dahliae to Bacillus and biocontrol bacteria.Results: By using in vitro confrontation bioassays, we found that under the stress induced by Bacillus, the spore vitality of V. dahliae with larger colonies was higher, and more abundant microsclerotia were formed. Then, according to the RNA-Seq analysis, the target of rapamycin (TOR) and mitophagy pathways were enriched among the significantly upregulated 542 genes observed in two co-culture groups with different colony sizes. In addition, in the group of V. dahliae with large colonies, the pathways related to cell wall synthesis, microsclerotia formation and the clearance of reactive oxygen species were regulated, and the expression of genes was up-regulated.Conclusion: This study found that the larger colonies of V. dahliae were more resistant to the antagonistic actions of Bacillus and the likelihood of the formation of homeostasis. Therefore, the prevention of Verticillium wilt by Bacillus is more effective than the treatment of an active fungal infection. These transcriptomic insights provide direction for the use of fungicides in the prevention and treatment of diseases such as Verticillium wilt.

scholarly journals SUMOylation in Human Pathogenic Fungi: Role in Physiology and Virulence

2020 ◽  
Vol 6 (1) ◽  
pp. 32
Author(s):  
Mahima Sagar Sahu ◽  
Sandip Patra ◽  
Kundan Kumar ◽  
Rupinder Kaur
Keyword(s):  
Protein Modification ◽  
Sequence Similarity ◽  
In Silico Analysis ◽  
Pathogenic Fungi ◽  
Mammalian Host ◽  
Nuclear Pore ◽  
Yeast Saccharomyces Cerevisiae ◽  
Pathogenic Yeast ◽  
Sumo Conjugation

The small ubiquitin-related modifier (SUMO) protein is an important component of the post-translational protein modification systems in eukaryotic cells. It is known to modify hundreds of proteins involved in diverse cellular processes, ranging from nuclear pore dynamics to signal transduction pathways. Owing to its reversible nature, the SUMO-conjugation of proteins (SUMOylation) holds a prominent place among mechanisms that regulate the functions of a wide array of cellular proteins. The dysfunctional SUMOylation system has been associated with many human diseases, including neurodegenerative and autoimmune disorders. Furthermore, the non-pathogenic yeast Saccharomyces cerevisiae has served as an excellent model to advance our understanding of enzymes involved in SUMOylation and proteins modified by SUMOylation. Taking advantage of the tools and knowledge obtained from the S. cerevisiae SUMOylation system, research on fungal SUMOylation is beginning to gather pace, and new insights into the role of SUMOylation in the pathobiology of medically important fungi are emerging. Here, we summarize the known information on components of the SUMOylation machinery, and consequences of overexpression or deletion of these components in the human pathogenic fungi, with major focus on two prevalent Candida bloodstream pathogens, C. albicans and C. glabrata. Additionally, we have identified SUMOylation components, through in silico analysis, in four medically relevant fungi, and compared their sequence similarity with S. cerevisiae counterparts. SUMOylation modulates the virulence of C. albicans and C. glabrata, while it is required for conidia production in Aspergillus nidulans and A. flavus. In addition to highlighting these recent developments, we discuss how SUMOylation fine tunes the expression of virulence factors, and influences survival of fungal cells under diverse stresses in vitro and in the mammalian host.

Evaluation of the in vitro and in vivo dimorphism ofSporothrix schenckii,Blastomyces dermatitidis, andParacoccidioides brasiliensisisolates after preservation in mineral oil

2004 ◽  
Vol 50 (6) ◽  
pp. 445-449 ◽  
Author(s):  
Renata Ferretti de Lima ◽  
Marcelly Maria dos Santos Brito ◽  
Guido Manoel Vidal Schäffer ◽  
Osana Cunha de Lima ◽  
Cintia de Moraes Borba
Keyword(s):  
Fungal Infections ◽  
Paracoccidioides Brasiliensis ◽  
Sporothrix Schenckii ◽  
Brain Heart Infusion ◽  
Morphological Differentiation ◽  
Pathogenic Fungi ◽  
Mineral Oil ◽  
Blastomyces Dermatitidis

Morphological differentiation has commanded attention for its putative impact on the pathogenesis of invasive fungal infections. We evaluated in vitro and in vivo the dimorphism from mycelial to yeast-phase of Sporothrix schenckii, Blastomyces dermatitidis and Paracoccidioides brasiliensis isolates, two strains for each species, preserved in mineral oil. S. schenckii strains showed typical micromorphology at 25 °C but one strain was unable to complete the dimorphic process in vitro. After in vivo passage through mice the strains had the ability to turn into yeast-like cells and to form colonies on brain-heart infusion medium at 36 °C. B. dermatitidis strains grew as dirty white to brownish membranous colonies at 25 °C and their micromorphology showed thin filaments with single hyaline conidia. At 36 °C the colonies did not differ from those grown at 25 °C, but produced a transitional micromorphology. P. brasiliensis strains grew as cream-colored cerebriform colonies at 25 °C showing a transitional morphology. B. dermatitidis and P. brasiliensis strains did not turn into yeast-like cells in vivo. The present results demonstrate that B. dermatitidis and P. brasiliensis strains were unable to complete the dimorphic process even after in vivo passage, in contrast to the S. schenckii strain.Key words: pathogenic fungi, in vitro storage, in vivo passage, morphogenesis.

scholarly journals Extracellular vesicles from Paracoccidioides brasiliensis induced M1 polarization in vitro

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Thiago Aparecido da Silva ◽  
Maria Cristina Roque-Barreira ◽  
Arturo Casadevall ◽  
Fausto Almeida
Keyword(s):  
Extracellular Vesicles ◽  
Paracoccidioides Brasiliensis ◽  
M1 Polarization

scholarly journals Targeting the Homoserine Dehydrogenase of Paracoccidioides Species for Treatment of Systemic Fungal Infections

2017 ◽  
Vol 61 (9) ◽  
Author(s):  
Mariane C. Bagatin ◽  
Arethusa L. Pimentel ◽  
Débora C. Biavatti ◽  
Ernani A. Basso ◽  
Erika S. Kioshima ◽  
...  
Keyword(s):  
Tertiary Structure ◽  
Fungal Infections ◽  
Paracoccidioides Brasiliensis ◽  
Pathogenic Fungi ◽  
Dynamics Simulation ◽  
Homoserine Dehydrogenase ◽  
Content Type ◽  
Zinc Database ◽  
Etiological Agents

ABSTRACT This work evaluated new potential inhibitors of the enzyme homoserine dehydrogenase (HSD) of Paracoccidioides brasiliensis, one of the etiological agents of paracoccidioidomycosis. The tertiary structure of the protein bonded to the analogue NAD, and l-homoserine was modeled by homology. The model with the best output was subjected to gradient minimization, redocking, and molecular dynamics simulation. Virtual screening simulations with 187,841 molecules purchasable from the Zinc database were performed. After the screenings, 14 molecules were selected and analyzed by the use of absorption, distribution, metabolism, excretion, and toxicity criteria, resulting in four compounds for in vitro assays. The molecules HS1 and HS2 were promising, exhibiting MICs of 64 and 32 μg · ml−1, respectively, for the Pb18 isolate of P. brasilensis, 64 μg · ml−1 for two isolates of P. lutzii, and also synergy with itraconazole. The application of these molecules to human-pathogenic fungi confirmed that the HSD enzyme may be used as a target for the development of drugs with specific action against paracoccidioidomycosis; moreover, these compounds may serve as leads in the design of new antifungals.

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