Your browser doesn't support javascript.
Show: 20 | 50 | 100
Results 1 - 6 de 6
Filter
1.
Int J Mol Sci ; 22(16)2021 Aug 12.
Article in English | MEDLINE | ID: covidwho-1354987

ABSTRACT

Recently, we have experienced a serious pandemic. Despite significant technological advances in molecular technologies, it is very challenging to slow down the infection spread. It appeared that due to globalization, SARS-CoV-2 spread easily and adapted to new environments or geographical or weather zones. Additionally, new variants are emerging that show different infection potential and clinical outcomes. On the other hand, we have some experience with other pandemics and some solutions in virus elimination that could be adapted. This is of high importance since, as the latest reports demonstrate, vaccine technology might not follow the new, mutated virus outbreaks. Thus, identification of novel strategies and markers or diagnostic methods is highly necessary. For this reason, we present some of the latest views on SARS-CoV-2/COVID-19 therapeutic strategies and raise a solution based on miRNA. We believe that in the face of the rapidly increasing global situation and based on analogical studies of other viruses, the possibility of using the biological potential of miRNA technology is very promising. It could be used as a promising diagnostic and prognostic factor, as well as a therapeutic target and tool.


Subject(s)
COVID-19 Vaccines/administration & dosage , COVID-19/drug therapy , COVID-19/therapy , MicroRNAs/genetics , Angiotensin-Converting Enzyme 2/immunology , Antimalarials/pharmacology , COVID-19/genetics , COVID-19/immunology , COVID-19/virology , Cytokine Release Syndrome/immunology , Cytokine Release Syndrome/virology , Humans , Immunization, Passive , MicroRNAs/analysis , Pandemics , SARS-CoV-2/drug effects , SARS-CoV-2/genetics , SARS-CoV-2/immunology , Vitamins/pharmacology
2.
Biomolecules ; 11(7)2021 07 20.
Article in English | MEDLINE | ID: covidwho-1323104

ABSTRACT

We report the case of a 77-year-old woman affected by coronavirus disease-19 (COVID-19) who developed an occlusive arterial disease of the lower limb requiring a left leg amputation. We studied the mechanisms of vascular damage by SARS-CoV-2 by means of a comprehensive multi-technique in situ analysis on the diseased popliteal arterial district, including immunohistochemistry (IHC), transmission electron microscopy (TEM) and miRNA analysis. At histological analyses, we observed a lymphocytic inflammatory infiltrate, oedema and endothelialitis of adventitial vasa vasorum while the media was normal and the intima had only minor changes. The vasa vasorum expressed the ACE2 receptor and factor VIII; compared with the controls, VEGFR2 staining was reduced. TEM analyses showed endothelial injury and numerous Weibel-Palade bodies in the cytoplasm. No coronavirus particle was seen. IL-6 protein and mRNA, together with miR-155-5p and miRs-27a-5p, which can target IL-6, were significantly increased compared with that in the controls. Our case report suggests an involvement of adventitial artery microcirculation by inflammation in the course of COVID-19. Without evident signs of current infection by SARS-CoV-2, endothelial cells show a spectrum of structural and functional alterations that can fuel the cardiovascular complications observed in people infected with SARS-CoV-2.


Subject(s)
Arterial Occlusive Diseases/etiology , COVID-19/complications , Inflammation/etiology , Aged , Arterial Occlusive Diseases/pathology , COVID-19/pathology , Female , Humans , Inflammation/pathology , Leg/blood supply , Leg/pathology , MicroRNAs/analysis , Microcirculation , SARS-CoV-2/isolation & purification
3.
Cells ; 10(7)2021 07 15.
Article in English | MEDLINE | ID: covidwho-1314589

ABSTRACT

MicroRNAs are gene expression regulators associated with several human pathologies, including those generated by viral infections. Their role in SARS-CoV-2 infection and COVID-19 has been investigated and reviewed in many informative studies; however, a thorough miRNA outline in SARS-CoV-2-infected pregnant women (SIPW), at both systemic and placental levels, is missing. To fill this gap, blood and placenta biopsies collected at delivery from 15 asymptomatic SIPW were immediately analysed for: miRNA expression (n = 84) (QPCR array), antiviral/immune mRNA target expression (n = 74) (QGene) and cytokine/chemokines production (n = 27) (Multiplex ELISA). By comparing these results with those obtained from six uninfected pregnant women (UPW), we observed that, following SARS-CoV-2 infection, the transcriptomic profile of pregnant women is significantly altered in different anatomical districts, even in the absence of clinical symptoms and vertical transmission. This characteristic combination of miRNA and antiviral/immune factors seems to control both the infection and the dysfunctional immune reaction, thus representing a positive correlate of protection and a potential therapeutic target against SARS-CoV-2.


Subject(s)
COVID-19/genetics , MicroRNAs/genetics , Pregnancy Complications, Infectious/genetics , Adult , COVID-19/blood , COVID-19/diagnosis , Female , Humans , MicroRNAs/analysis , MicroRNAs/blood , Placenta/metabolism , Pregnancy , Pregnancy Complications, Infectious/blood , Pregnancy Complications, Infectious/diagnosis , SARS-CoV-2/isolation & purification , Transcriptome , Young Adult
4.
Cells ; 10(3)2021 03 04.
Article in English | MEDLINE | ID: covidwho-1125522

ABSTRACT

Since the outbreak of the COVID-19 crisis, the handling of biological samples from confirmed or suspected SARS-CoV-2-positive individuals demanded the use of inactivation protocols to ensure laboratory operators' safety. While not standardized, these practices can be roughly divided into two categories, namely heat inactivation and solvent-detergent treatments. These routine procedures should also apply to samples intended for Extracellular Vesicles (EVs) analysis. Assessing the impact of virus-inactivating pre-treatments is therefore of pivotal importance, given the well-known variability introduced by different pre-analytical steps on downstream EVs isolation and analysis. Arguably, shared guidelines on inactivation protocols tailored to best address EVs-specific requirements will be needed among the analytical community, yet deep investigations in this direction have not yet been reported. We here provide insights into SARS-CoV-2 inactivation practices to be adopted prior to serum EVs analysis by comparing solvent/detergent treatment vs. heat inactivation. Our analysis entails the evaluation of EVs recovery and purity along with biochemical, biophysical and biomolecular profiling by means of a set of complementary analytical techniques: Nanoparticle Tracking Analysis, Western Blotting, Atomic Force Microscopy, miRNA content (digital droplet PCR) and tetraspanin assessment by microarrays. Our data suggest an increase in ultracentrifugation (UC) recovery following heat treatment; however, it is accompanied by a marked enrichment in EVs-associated contaminants. On the other hand, solvent/detergent treatment is promising for small EVs (<150 nm range), yet a depletion of larger vesicular entities was detected. This work represents a first step towards the identification of optimal serum inactivation protocols targeted to EVs analysis.


Subject(s)
COVID-19/blood , Containment of Biohazards/methods , Extracellular Vesicles/chemistry , Virus Inactivation , COVID-19/virology , Detergents/pharmacology , Extracellular Vesicles/drug effects , Extracellular Vesicles/genetics , Hot Temperature , Humans , MicroRNAs/analysis , Microarray Analysis , Microscopy, Atomic Force , SARS-CoV-2 , Tetraspanins/analysis , Ultracentrifugation
5.
ACS Nano ; 15(1): 1167-1178, 2021 01 26.
Article in English | MEDLINE | ID: covidwho-1014984

ABSTRACT

Existing methods for RNA diagnostics, such as reverse transcription PCR (RT-PCR), mainly rely on nucleic acid amplification (NAA) and RT processes, which are known to introduce substantial issues, including amplification bias, cross-contamination, and sample loss. To address these problems, we introduce a confinement effect-inspired Cas13a assay for single-molecule RNA diagnostics, eliminating the need for NAA and RT. This assay involves confining the RNA-triggered Cas13a catalysis system in cell-like-sized reactors to enhance local concentrations of target and reporter simultaneously, via droplet microfluidics. It achieves >10 000-fold enhancement in sensitivity when compared to the bulk Cas13a assay and enables absolute digital single-molecule RNA quantitation. We experimentally demonstrate its broad applicability for precisely counting microRNAs, 16S rRNAs, and SARS-CoV-2 RNA from synthetic sequences to clinical samples with excellent accuracy. Notably, this direct RNA diagnostic technology enables detecting a wide range of RNA molecules at the single-molecule level. Moreover, its simplicity, universality, and excellent quantification capability might render it to be a dominant rival to RT-qPCR.


Subject(s)
CRISPR-Cas Systems , Microfluidics , RNA/analysis , Cell Line, Tumor , Enterococcus faecalis , Escherichia coli , Humans , Klebsiella pneumoniae , MCF-7 Cells , MicroRNAs/analysis , Pseudomonas aeruginosa , RNA, Ribosomal, 16S/analysis , Reverse Transcriptase Polymerase Chain Reaction , SARS-CoV-2/genetics , Staphylococcus aureus
6.
Med Hypotheses ; 143: 110124, 2020 Oct.
Article in English | MEDLINE | ID: covidwho-662554

ABSTRACT

Utilising biomarkers for COVID-19 diagnosis, prediction of treatment response and overall prognostication have been investigated recently. However, these ventures have only considered the use of blood-based molecular markers. Saliva is another biofluid that warrants being applied in similar fashion with major advantages that centres on its non-invasive and repeatable collection as well as cost-efficiency. To this end, this article presents a hypothesis for the sources of biomarkers useful clinically for COVID-19 disease outcome estimation and identify the likely implications of their detection in saliva.


Subject(s)
Betacoronavirus , Biomarkers/analysis , Coronavirus Infections/metabolism , Models, Immunological , Pandemics , Pneumonia, Viral/metabolism , Saliva/chemistry , Biomarkers/blood , COVID-19 , Coronavirus Infections/diagnosis , Coronavirus Infections/immunology , Coronavirus Infections/therapy , Cytokines/analysis , Diagnostic Tests, Routine , Extracellular Vesicles , Gingival Crevicular Fluid/chemistry , Humans , MicroRNAs/analysis , Mouth Diseases/complications , Mouth Diseases/metabolism , Pneumonia, Viral/diagnosis , Pneumonia, Viral/immunology , Pneumonia, Viral/therapy , SARS-CoV-2 , Saliva/immunology , Saliva/virology , Salivary Glands/metabolism , Salivary Glands/virology , Salivary Proteins and Peptides/analysis
SELECTION OF CITATIONS
SEARCH DETAIL