Your browser doesn't support javascript.
Show: 20 | 50 | 100
Results 1 - 20 de 253
Filter
Add filters

Document Type
Year range
1.
Biosens Bioelectron ; 196: 113701, 2022 Jan 15.
Article in English | MEDLINE | ID: covidwho-1638371

ABSTRACT

Although CRISPR-Cas12a and CRISPR-Cas13a systems work individually effective on gene detection, their multiplex detection capability is limited due to the lack of specific probe cleavage mechanism. Herein we present a high-efficient dual-gene diagnostic technique based on the orthogonal DNA/RNA collateral cleavage mechanism of Cas12a/Cas13a system. In this design, dual-gene amplified products from the multiplex recombinase polymerase amplification (RPA) were simultaneously detected by Cas12a and Cas13a assay in a single tube. The resulting orthogonal DNA/RNA collateral cleavage can specifically illuminate two spectral differentiated DNA and RNA probes, respectively. By integrating with the smartphone-based fluorescence readout, a portable detection platform is achieved. As a proof-of-concept, reliable dual-gene detection of SARS-CoV-2 and African Swine fever virus (ASFV) were demonstrated, exhibiting 100% sensitivity and specificity for clinical samples analysis (32 swab specimens for SARS-CoV-2 and 35 ASFV suspected swine blood samples). This developed portable dual-gene detection platform can provide accurate point-of-care screening of infectious diseases in resources-limited settings.


Subject(s)
African Swine Fever Virus , Biosensing Techniques , COVID-19 , Animals , CRISPR-Cas Systems/genetics , Humans , SARS-CoV-2 , Swine
2.
Sci Rep ; 12(1): 696, 2022 01 13.
Article in English | MEDLINE | ID: covidwho-1621270

ABSTRACT

Despite encouraging preclinical data, therapies to reduce ARDS mortality remains a globally unmet need, including during the COVID-19 pandemic. We previously identified extracellular nicotinamide phosphoribosyltransferase (eNAMPT) as a novel damage-associated molecular pattern protein (DAMP) via TLR4 ligation which regulates inflammatory cascade activation. eNAMPT is tightly linked to human ARDS by biomarker and genotyping studies in ARDS subjects. We now hypothesize that an eNAMPT-neutralizing mAb will significantly reduce the severity of ARDS lung inflammatory lung injury in diverse preclinical rat and porcine models. Sprague Dawley rats received eNAMPT mAb intravenously following exposure to intratracheal lipopolysaccharide (LPS) or to a traumatic blast (125 kPa) but prior to initiation of ventilator-induced lung injury (VILI) (4 h). Yucatan minipigs received intravenous eNAMPT mAb 2 h after initiation of septic shock and VILI (12 h). Each rat/porcine ARDS/VILI model was strongly associated with evidence of severe inflammatory lung injury with NFkB pathway activation and marked dysregulation of the Akt/mTORC2 signaling pathway. eNAMPT neutralization dramatically reduced inflammatory indices and the severity of lung injury in each rat/porcine ARDS/VILI model (~ 50% reduction) including reduction in serum lactate, and plasma levels of eNAMPT, IL-6, TNFα and Ang-2. The eNAMPT mAb further rectified NFkB pathway activation and preserved the Akt/mTORC2 signaling pathway. These results strongly support targeting the eNAMPT/TLR4 inflammatory pathway as a potential ARDS strategy to reduce inflammatory lung injury and ARDS mortality.


Subject(s)
Acute Chest Syndrome/metabolism , Mechanistic Target of Rapamycin Complex 2/metabolism , NF-kappa B/metabolism , Nicotinamide Phosphoribosyltransferase/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction/physiology , Animals , Antibodies, Neutralizing/metabolism , Biomarkers/metabolism , COVID-19/metabolism , Disease Models, Animal , Inflammation/metabolism , Lipopolysaccharides/metabolism , Lung/metabolism , Male , Rats , Rats, Sprague-Dawley , SARS-CoV-2/pathogenicity , Swine
3.
BMJ Case Rep ; 15(1)2022 Jan 07.
Article in English | MEDLINE | ID: covidwho-1612961

ABSTRACT

Postoperative fevers are common in hospitalised patients and warrant workup beyond the early post-op period. A 50-year-old man was admitted after sustaining a tibial plateau fracture. Fevers began 3 days after external fixation and persisted through a second surgery despite initial negative workup. Careful review of medications revealed enoxaparin as the instigating agent of a febrile drug reaction, and the fevers resolved after discontinuing the drug. On further questioning, it was discovered the patient had an allergy to pork, from which the main components of enoxaparin are typically derived. To our knowledge, this is the first reported enoxaparin-induced fever in the setting of a pork allergy. Enoxaparin-induced fevers should be considered in patients with unexplained post-op fever. Our case demonstrates the importance of analysing newly administered medications. Simple detailed history may significantly reduce patient morbidity and help to broaden differentials during investigation.


Subject(s)
Fever of Unknown Origin , Hypersensitivity , Pork Meat , Red Meat , Animals , Anticoagulants/adverse effects , Enoxaparin/adverse effects , Humans , Male , Middle Aged , Postoperative Complications/chemically induced , Swine
4.
Sensors (Basel) ; 21(24)2021 Dec 08.
Article in English | MEDLINE | ID: covidwho-1592974

ABSTRACT

The encroachment of wild boars into urban areas is a growing problem. The occurrence of wild boars in cities leads to conflict situations. Socio-spatial conflicts can escalate to a varied degree. Assessments of these conflicts can be performed by analyzing spatial data concerning the affected locations and wild boar behaviors. The collection of spatial data is a laborious and costly process that requires access to urban surveillance systems, in addition to regular analyses of intervention reports. A supporting method for assessing the risk of wild boar encroachment and socio-spatial conflict in cities was proposed in the present study. The developed approach relies on big data, namely, multimedia and descriptive data that are on social media. The proposed method was tested in the city of Olsztyn in Poland. The main aim of this study was to evaluate the applicability of data crowdsourced from a popular social networking site for determining the location and severity of conflicts. A photointerpretation method and the kernel density estimation (KDE) tool implemented in ArcGIS Desktop 10.7.1 software were applied in the study. The proposed approach fills a gap in the application of crowdsourcing data to identify types of socio-spatial conflicts involving wild boars in urban areas. Validation of the results with reports of calls to intervention services showed the high coverage of this approach and thus the usefulness of crowdsourcing data.


Subject(s)
Social Media , Sus scrofa , Animals , Cities , Humans , Poland , Spatial Analysis , Swine
5.
Front Public Health ; 9: 765501, 2021.
Article in English | MEDLINE | ID: covidwho-1591734

ABSTRACT

Introduction: During the ongoing COVID-19 pandemic there have been much publicised shortages in Personal Protective Equipment for frontline health care workers, from masks to gowns. Recent previous airborne pandemics provide an opportunity to learn how to effectively lead and manage supply chains during crisis situations. Identifying and plotting this learning against time will reveal what has been learnt, when and, significantly, what can be learnt for the future. Aims: (i) To identify the temporal trajectory of leadership and management learning in health supply chain management through pandemics and (ii) to identify leadership and management lessons to enable the resilient supply of key items such as PPE in future pandemics. Methods: We undertook a scoping review in line with PRISMA (scoping review extension) searching Business Source Premier, Health Business Elite, Medline, ProQuest Business Collection and PubMed. Search terms were focused on recent airborne pandemics (SARS; Ebola; Zika virus; H1N1 swine flu, COVID-19), supply chain management, PPE, leadership, learning, inhibitors and facilitators and resilience e.g., SARS AND supply chain* AND ("personal protective equipment" OR PPE) (leaders* OR manage*) Titles and abstracts were downloaded to Endnote and duplicates removed. Two authors independently screened all of the titles and abstracts. Inclusion criteria focused on leadership and management in health supply chains during pandemics, peer reviewed or grey literature (either from business journals or reports): exclusion criteria included not in English and not focused on a named pandemic. Once interrater reliability was assured, authors completed a title and abstract screening independently. Ten percent of the resultant full text articles were screened by both authors, once agreement was reached the full text articles were screened independently noting reasons for exclusion. A data extraction tool was designed to capture findings from the final articles included in the review. Results/Discussion: We found 92 articles and, after screening, included 30 full text articles. The majority were focused on COVID-19 (N = 27) and most were from the USA (N = 13). We identified four themes related to leadership and management of pandemic PPE supply chains, (i) Leadership and management learning for pandemic PPE supply chain management, (ii) Inhibitors of PPE supply chain resilience during a pandemic, (iii) Facilitators employed to manage the immediate impacts of PPE supply chain demands during a pandemic,and (iv) Facilitators proposed to ensure longer term resilience of PPE supply chains during pandemics Our study suggests there has been limited leadership and management learning for PPE supply chains from previous pandemics, however there has been extensive learning through the COVID-19 pandemic. Lessons included the importance of planning, the significance of collaboration and relationship building. Resilience of PPE supply chains was reported to be dependent on multiple levels from individuals to organisation level and also interdependent on (i) sustainability, (ii) the practise of PPE and (iii) long term environmental impact of PPE suggesting the need, long term, to move to a circular economy approach.


Subject(s)
COVID-19 , Influenza A Virus, H1N1 Subtype , Zika Virus Infection , Zika Virus , Animals , Delivery of Health Care , Humans , Leadership , Pandemics/prevention & control , Personal Protective Equipment , Reproducibility of Results , SARS-CoV-2 , Swine
6.
PLoS One ; 16(12): e0252972, 2021.
Article in English | MEDLINE | ID: covidwho-1598722

ABSTRACT

SARS-CoV-2 has clearly shown that efficient management of infectious diseases requires a top-down approach which must be complemented with a bottom-up response to be effective. Here we investigate a novel approach to surveillance for transboundary animal diseases using African Swine (ASF) fever as a model. We collected data both at a population level and at the local level on information-seeking behavior respectively through digital data and targeted questionnaire-based surveys to relevant stakeholders such as pig farmers and veterinary authorities. Our study shows how information-seeking behavior and resulting public attention during an epidemic, can be identified through novel data streams from digital platforms such as Wikipedia. Leveraging attention in a critical moment can be key to providing the correct information at the right moment, especially to an interested cohort of people. We also bring evidence on how field surveys aimed at local workers and veterinary authorities remain a crucial tool to assess more in-depth preparedness and awareness among front-line actors. We conclude that these two tools should be used in combination to maximize the outcome of surveillance and prevention activities for selected transboundary animal diseases such as ASF.


Subject(s)
African Swine Fever/epidemiology , Epidemics/prevention & control , Epidemiological Monitoring , Livestock/virology , Animals , Awareness , Estonia/epidemiology , Farmers , Internet , Statistics, Nonparametric , Surveys and Questionnaires , Swine
7.
J Environ Manage ; 304: 114296, 2022 Feb 15.
Article in English | MEDLINE | ID: covidwho-1568834

ABSTRACT

Wastewater-based epidemiology (WBE) for severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2) is a powerful tool to complement syndromic surveillance. Although detection of SARS-CoV-2 in raw wastewater may be prompted with good recoveries during periods of high community prevalence, in the early stages of population outbreaks concentration procedures are required to overcome low viral concentrations. Several methods have become available for the recovery of SARS-CoV-2 from raw wastewater, generally involving filtration. However, these methods are limited to small sample volumes, possibly missing the early stages of virus circulation, and restrained applicability across different water matrices. The aim of this study was thus to evaluate the performance of three methods enabling the concentration of SARS-CoV-2 from large volumes of wastewater: i) hollow fiber filtration using the inuvai R180, with an enhanced elution protocol and polyethylene glycol (PEG) precipitation; ii) PEG precipitation; and iii) skimmed milk flocculation. The performance of the three approaches was evaluated in wastewater from multiple wastewater treatment plants (WWTP) with distinct singularities, according to: i) effective volume; ii) percentage of recovery; iii) extraction efficiency; iv) inhibitory effect; and v) the limits of detection and quantification. The inuvai R180 system had the best performance, with detection of spiked control across all samples, with average recovery percentages of 68% for porcine epidemic diarrhea virus (PEDV), with low variability. Mean recoveries for PEG precipitation and skimmed milk flocculation were 9% and 14%, respectively. The inuvai R180 enables the scalability of volumes without negative impact on the costs, time for analysis, and recovery/inhibition. Moreover, hollow fiber ultrafilters favor the concentration of different microbial taxonomic groups. Such combined features make this technology attractive for usage in environmental waters monitoring.


Subject(s)
COVID-19 , Viruses , Animals , Humans , SARS-CoV-2 , Swine , Waste Water
8.
Vet Microbiol ; 264: 109299, 2022 Jan.
Article in English | MEDLINE | ID: covidwho-1559479

ABSTRACT

Porcine deltacoronavirus (PDCoV) is an emerging porcine enteric coronavirus that causes severe diarrhea in piglets and results in serious economic losses. There are no effective vaccines and antiviral drugs to prevent and treat PDCoV infection currently. Griffithsin (GRFT) is a lectin with potent antiviral activity against enveloped viruses because of its ability to specifically bind N-linked high-mannose oligosaccharides. GRFT has been reported to possess antiviral activity against severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), and porcine epidemic diarrhea virus (PEDV). Here, we first confirmed the antiviral activity of GRFT against PDCoV in vitro. The infected cells (%) and virus titers were significantly decreased at concentration 1 µg/mL or above of GRFT. Time-course experiments revealed that GRFT inhibits PDCoV infection at the adsorption and penetration step. GRFT binding to PDCoV spike (S) protein on the surface wraps the virus and blocks its entry. The outstanding antiviral potency indicates that GRFT has the potential value as a candidate drug for the prevention and treatment of PDCoV infection.


Subject(s)
Deltacoronavirus , Plant Lectins , Animals , Antiviral Agents/pharmacology , Cell Culture Techniques/veterinary , Coronavirus Infections/drug therapy , Coronavirus Infections/veterinary , Deltacoronavirus/drug effects , Plant Lectins/pharmacology , Swine , Swine Diseases/drug therapy
9.
Viruses ; 13(12)2021 12 06.
Article in English | MEDLINE | ID: covidwho-1555020

ABSTRACT

Porcine deltacoronavirus (PDCoV) is a novel coronavirus that causes diarrhea in nursing piglets. Studies showed that PDCoV uses porcine aminopeptidase N (pAPN) as an entry receptor, but the infection of pAPN-knockout cells or pigs with PDCoV revealed that pAPN might be not a critical functional receptor, implying there exists an unidentified receptor involved in PDCoV infection. Herein, we report that sialic acid (SA) can act as an attachment receptor for PDCoV invasion and facilitate its infection. We first demonstrated that the carbohydrates destroyed on the cell membrane using NaIO4 can alleviate the susceptibility of cells to PDCoV. Further study showed that the removal of SA, a typical cell-surface carbohydrate, could influence the PDCoV infectivity to the cells significantly, suggesting that SA was involved in the infection. The results of plaque assay and Western blotting revealed that SA promoted PDCoV infection by increasing the number of viruses binding to SA on the cell surface during the adsorption phase, which was also confirmed by atomic force microscopy at the microscopic level. In in vivo experiments, we found that the distribution levels of PDCoV and SA were closely relevant in the swine intestine, which contains huge amount of trypsin. We further confirmed that SA-binding capacity to PDCoV is related to the pre-treatment of PDCoV with trypsin. In conclusion, SA is a novel attachment receptor for PDCoV infection to enhance its attachment to cells, which is dependent on the pre-treatment of trypsin on PDCoV. This study paves the way for dissecting the mechanisms of PDCoV-host interactions and provides new strategies to control PDCoV infection.


Subject(s)
Deltacoronavirus/physiology , N-Acetylneuraminic Acid/metabolism , Receptors, Virus/metabolism , Trypsin/metabolism , Virus Attachment , Animals , Carbohydrates , Cell Line , Cell Membrane/metabolism , Cell Membrane/virology , Coronavirus Infections/veterinary , Coronavirus Infections/virology , Deltacoronavirus/drug effects , Host-Pathogen Interactions , Intestines/metabolism , Intestines/virology , Periodic Acid/pharmacology , Swine , Swine Diseases/virology , Trypsin/pharmacology
10.
Transbound Emerg Dis ; 68(6): 3194-3199, 2021 Nov.
Article in English | MEDLINE | ID: covidwho-1532923

ABSTRACT

African swine fever (ASF) is a contagious haemorrhagic disease in pigs and has become endemic in several Vietnam provinces since the first outbreak in 2019. The presence of carriers and the recurrence of disease in the surviving swine herd after an ASF outbreak has not previously been properly evaluated. In this study, pigs naturally infected with an acute form of ASF were allowed to recover from the disease. A serological follow-up was conducted for more than 14 months with 14 convalescent gilts and their offspring. All convalescent animals had long lasting high serum antibody levels without persistent viremia. They also did not excrete virus via nasal discharge post-recovery. These convalescent pigs could partially perform as replacement gilts despite the fact that ASF affected reproductive performance. Here, we confirmed that there were neither the carriers of nor recurrence of disease in the convalescent pigs and their offspring following the outbreak of acute ASF. These findings may facilitate efforts to design a new farming model in ASF endemic provinces in Vietnam where there is a lack of a repopulation strategy due to the limited funding received from the local regulatory authorities.


Subject(s)
African Swine Fever Virus , African Swine Fever , Swine Diseases , African Swine Fever/epidemiology , African Swine Fever Virus/genetics , Animals , Disease Outbreaks/veterinary , Female , Follow-Up Studies , Sus scrofa , Swine , Vietnam/epidemiology
11.
Transbound Emerg Dis ; 68(6): 3349-3359, 2021 Nov.
Article in English | MEDLINE | ID: covidwho-1526423

ABSTRACT

The influenza D virus (IDV) was first identified and characterized in 2011. Considering the virus' zoonotic potential, its genome nature (segmented RNA virus), its worldwide circulation in livestock and its role in bovine respiratory disease, an increased interest is given to IDV. However, few data are available on drivers of emergence of IDV. We first listed fifty possible drivers of emergence of IDV in ruminants and swine. As recently carried out for COVID-19 in pets (Transboundary and Emerging Diseases, 2020), a scoring system was developed per driver and scientific experts (N = 28) were elicited to (a) allocate a score to each driver, (b) weight the drivers' scores within each domain and (c) weight the different domains among themselves. An overall weighted score was calculated per driver, and drivers were ranked in decreasing order. Drivers with comparable likelihoods to play a role in the emergence of IDV in ruminants and swine in Europe were grouped using a regression tree analysis. Finally, the robustness of the expert elicitation was verified. Eight drivers were ranked with the highest probability to play a key role in the emergence of IDV: current species specificity of the causing agent of the disease; influence of (il)legal movements of live animals (ruminants, swine) from neighbouring/European Union member states and from third countries for the disease to (re-)emerge in a given country; detection of emergence; current knowledge of the pathogen; vaccine availability; animal density; and transport vehicles of live animals. As there is still limited scientific knowledge on the topic, expert elicitation of knowledge and multi-criteria decision analysis, in addition to clustering and sensitivity analyses, are very important to prioritize future studies, starting from the top eight drivers. The present methodology could be applied to other emerging animal diseases.


Subject(s)
COVID-19 , Influenza, Human , Orthomyxoviridae Infections , Swine Diseases , Animals , COVID-19/veterinary , Cattle , Europe/epidemiology , Humans , Orthomyxoviridae Infections/epidemiology , Orthomyxoviridae Infections/prevention & control , Orthomyxoviridae Infections/veterinary , SARS-CoV-2 , Swine , Swine Diseases/epidemiology , Swine Diseases/prevention & control
12.
Parasit Vectors ; 14(1): 282, 2021 May 26.
Article in English | MEDLINE | ID: covidwho-1523322

ABSTRACT

Trichinellosis is a foodborne disease caused by several Trichinella species around the world. In Chile, the domestic cycle was fairly well-studied in previous decades, but has been neglected in recent years. The aims of this study were to analyze, geographically, the incidence of trichinellosis in Chile to assess the relative risk and to analyze the incidence rate fluctuation in the last decades. Using temporal data spanning 1964-2019, as well as geographical data from 2010 to 2019, the time series of cases was analyzed with ARIMA models to explore trends and periodicity. The Dickey-Fuller test was used to study trends, and the Portmanteau test was used to study white noise in the model residuals. The Besag-York-Mollie (BYM) model was used to create Bayesian maps of the level of risk relative to that expected by the overall population. The association of the relative risk with the number of farmed swine was assessed with Spearman's correlation. The number of annual cases varied between 5 and 220 (mean: 65.13); the annual rate of reported cases varied between 0.03 and 1.9 cases per 105 inhabitants (mean: 0.53). The cases of trichinellosis in Chile showed a downward trend that has become more evident since the 1980s. No periodicities were detected via the autocorrelation function. Communes (the smallest geographical administrative subdivision) with high incidence rates and high relative risk were mostly observed in the Araucanía region. The relative risk of the commune was significantly associated with the number of farmed pigs and boar (Sus scrofa Linnaeus, 1758). The results allowed us to state that trichinellosis is not a (re)emerging disease in Chile, but the severe economic poverty rate of the Mapuche Indigenous peoples and the high number of backyard and free-ranging pigs seem to be associated with the high risk of trichinellosis in the Araucanía region.


Subject(s)
Swine Diseases/epidemiology , Trichinellosis/epidemiology , Animals , Bayes Theorem , Chile/epidemiology , Disease Outbreaks , Geographic Mapping , History, 20th Century , History, 21st Century , Incidence , Risk Assessment , Swine , Trichinella , Trichinellosis/history
13.
Parasit Vectors ; 14(1): 248, 2021 May 10.
Article in English | MEDLINE | ID: covidwho-1506030

ABSTRACT

BACKGROUND: Swine coccidiosis, a protozoan disease caused by coccidia, can result in diarrhoea and weight loss in piglets and even economic losses in the pig industry. Here, we report the first systematic review and meta-analysis of the prevalence of coccidia (including Eimeria spp. and Cystoisospora suis) in pigs in China. METHODS: Five databases (PubMed, ScienceDirect, Chinese Web of Knowledge, Wanfang, and Chongqing VIP) were searched and 50 studies (46,926 domestic pigs, 22 provinces) ultimately identified pertaining to the prevalence of coccidia infection from 1980 to 2019. We incorporated the effect size using the random-effects model in the "meta" package in R software and conducted univariate and multivariate meta-regression analyses using a mixed-effects model. RESULTS: The pooled prevalence rate of coccidia in pigs was 21.9%, including the C. suis infection rate of 9.1%. The highest prevalence of coccidia (39.6%) was found in northwest China, and this region also presented the lowest prevalence of C. suis (4.7%). In the subgroup analysis based on sampling year, the highest prevalence of coccidia was detected in 2001 or earlier (32.6%), whereas the lowest rate was found in 2012 or later (14.3%). An opposite trend was observed for C. suis (5.5% in 2000 or earlier vs 14.4% in 2000 or later). The prevalence of coccidia in extensive farming systems (29.5%) was higher than that in intensive farming systems (17.3%). In contrast, the point estimate of C. suis prevalence was lower in the extensive farming systems (5.1%) than in the intensive farming systems (10.0%), but the difference was not significant (P > 0.05). Among the four age categories, the highest total coccidia prevalence (26.2%) was found in finishing pigs, followed by suckling piglets (19.9%), whereas the highest prevalence of C. suis (14.9%) was observed in suckling piglets. CONCLUSIONS: Our findings suggest that coccidia infection in Chinese pigs is common, although the prevalence of C. suis in pigs does not receive sufficient attention. We recommend the rational use of anticoccidial drugs to avoid drug resistance and the development of preventive and control measures for C. suis to reduce the incidence of swine coccidiosis.


Subject(s)
Coccidiosis/epidemiology , Swine Diseases/epidemiology , Animals , China/epidemiology , Coccidia/classification , Coccidia/genetics , Coccidia/isolation & purification , Coccidia/physiology , Coccidiosis/parasitology , Feces/parasitology , Prevalence , Swine , Swine Diseases/parasitology
14.
Sci Rep ; 11(1): 21723, 2021 11 05.
Article in English | MEDLINE | ID: covidwho-1503978

ABSTRACT

Coronavirus with intact infectivity attached to PPE surfaces pose significant threat to the spread of COVID-19. We tested the hypothesis that an electroceutical fabric, generating weak potential difference of 0.5 V, disrupts the infectivity of coronavirus upon contact by destabilizing the electrokinetic properties of the virion. Porcine respiratory coronavirus AR310 particles (105) were placed in direct contact with the fabric for 1 or 5 min. Following one minute of contact, zeta potential of the porcine coronavirus was significantly lowered indicating destabilization of its electrokinetic properties. Size-distribution plot showed appearance of aggregation of the virus. Testing of the cytopathic effects of the virus showed eradication of infectivity as quantitatively assessed by PI-calcein and MTT cell viability tests. This work provides the rationale to consider the studied electroceutical fabric, or other materials with comparable property, as material of choice for the development of PPE in the fight against COVID-19.


Subject(s)
COVID-19/prevention & control , COVID-19/transmission , Electrochemistry/methods , Textiles , Animals , Anti-Infective Agents , Body Fluids , Cell Line , Cell Survival , Fluoresceins , Humans , Hydrogen Peroxide , Kinetics , Nanoparticles , Propidium , SARS-CoV-2 , Swine , Temperature , Tetrazolium Salts , Thiazoles , Virion , Wound Healing
15.
J Am Soc Nephrol ; 32(1): 86-97, 2021 01.
Article in English | MEDLINE | ID: covidwho-1496654

ABSTRACT

BACKGROUND: Cultured cell lines are widely used for research in the physiology, pathophysiology, toxicology, and pharmacology of the renal proximal tubule. The lines that are most appropriate for a given use depend upon the genes expressed. New tools for transcriptomic profiling using RNA sequencing (RNA-Seq) make it possible to catalog expressed genes in each cell line. METHODS: Fourteen different proximal tubule cell lines, representing six species, were grown on permeable supports under conditions specific for the respective lines. RNA-Seq followed standard procedures. RESULTS: Transcripts expressed in cell lines variably matched transcripts selectively expressed in native proximal tubule. Opossum kidney (OK) cells displayed the highest percentage match (45% of proximal marker genes [TPM threshold =15]), with pig kidney cells (LLC-PK1) close behind (39%). Lower-percentage matches were seen for various human lines, including HK-2 (26%), and lines from rodent kidneys, such as NRK-52E (23%). Nominally, identical OK cells from different sources differed substantially in expression of proximal tubule markers. Mapping cell line transcriptomes to gene sets for various proximal tubule functions (sodium and water transport, protein transport, metabolic functions, endocrine functions) showed that different lines may be optimal for experimentally modeling each function. An online resource (https://esbl.nhlbi.nih.gov/JBrowse/KCT/) has been created to interrogate cell line transcriptome data. Proteomic analysis of NRK-52E cells confirmed low expression of many proximal tubule marker proteins. CONCLUSIONS: No cell line fully matched the transcriptome of native proximal tubule cells. However, some of the lines tested are suitable for the study of particular metabolic and transport processes seen in the proximal tubule.


Subject(s)
Cell Culture Techniques/methods , Kidney Tubules, Proximal/metabolism , Transcriptome , Animals , Biological Transport , Cell Line , Chromatography, Liquid , Gene Expression Profiling , Humans , Internet , Mice , Opossums , Proteomics , RNA-Seq , Rats , Sequence Analysis, RNA , Species Specificity , Swine , Tandem Mass Spectrometry
16.
mBio ; 12(5): e0234221, 2021 10 26.
Article in English | MEDLINE | ID: covidwho-1494971

ABSTRACT

The recent emergence and spread of zoonotic viruses highlights that animal-sourced viruses are the biggest threat to global public health. Swine acute diarrhea syndrome coronavirus (SADS-CoV) is an HKU2-related bat coronavirus that was spilled over from Rhinolophus bats to swine, causing large-scale outbreaks of severe diarrhea disease in piglets in China. Unlike other porcine coronaviruses, SADS-CoV possesses broad species tissue tropism, including primary human cells, implying a significant risk of cross-species spillover. To explore host dependency factors for SADS-CoV as therapeutic targets, we employed genome-wide CRISPR knockout library screening in HeLa cells. Consistent with two independent screens, we identified the zinc finger DHHC-type palmitoyltransferase 17 (ZDHHC17 or ZD17) as an important host factor for SADS-CoV infection. Through truncation mutagenesis, we demonstrated that the DHHC domain of ZD17 that is involved in palmitoylation is important for SADS-CoV infection. Mechanistic studies revealed that ZD17 is required for SADS-CoV genomic RNA replication. Treatment of infected cells with the palmitoylation inhibitor 2-bromopalmitate (2-BP) significantly suppressed SADS-CoV infection. Our findings provide insight on SADS-CoV-host interactions and a potential therapeutic application. IMPORTANCE The recent emergence of deadly zoonotic viral diseases, including Ebola virus and SARS-CoV-2, emphasizes the importance of pandemic preparedness for the animal-sourced viruses with potential risk of animal-to-human spillover. Over the last 2 decades, three significant coronaviruses of bat origin, SARS-CoV, MERS-CoV, and SARS-CoV-2, have caused millions of deaths with significant economy and public health impacts. Lack of effective therapeutics against these coronaviruses was one of the contributing factors to such losses. Although SADS-CoV, another coronavirus of bat origin, was only known to cause fatal diarrhea disease in piglets, the ability to infect cells derived from multiple species, including human, highlights the potential risk of animal-to-human spillover. As part of our effort in pandemic preparedness, we explore SADS-CoV host dependency factors as targets for host-directed therapeutic development and found zinc finger DHHC-type palmitoyltransferase 17 is a promising drug target against SADS-CoV replication. We also demonstrated that a palmitoylation inhibitor, 2-bromopalmitate (2-BP), can be used as an inhibitor for SADS-CoV treatment.


Subject(s)
Acyltransferases/metabolism , Adaptor Proteins, Signal Transducing/metabolism , Alphacoronavirus/pathogenicity , Nerve Tissue Proteins/metabolism , Acyltransferases/genetics , Adaptor Proteins, Signal Transducing/genetics , Alphacoronavirus/drug effects , Animals , COVID-19/metabolism , HeLa Cells , Humans , Middle East Respiratory Syndrome Coronavirus/drug effects , Middle East Respiratory Syndrome Coronavirus/pathogenicity , Nerve Tissue Proteins/genetics , Palmitates/pharmacology , SARS Virus/drug effects , SARS Virus/pathogenicity , SARS-CoV-2/drug effects , SARS-CoV-2/pathogenicity , Swine
17.
Mater Sci Eng C Mater Biol Appl ; 116: 111260, 2020 Nov.
Article in English | MEDLINE | ID: covidwho-1452344

ABSTRACT

Polymeric nanoparticulate systems allow the encapsulation of bio-active substances, giving them protection against external agents and increasing the drug's bioavailability. The use of biocompatible and biodegradable polymers usually guarantees the harmless character of the formulation, and a controlled drug release is also assured. A relatively easy procedure to obtain polymeric formulations of bioactive agents is ionotropic gelation, which allows the synthesis of chitosan (CS) - sodium tri-polyphosphate nanoparticles (NPs) loading encapsulated proteins. In this work, Bovine serum albumin (BSA) model protein and a recombinant porcine alpha interferon variant were used to obtain nanoparticulate formulations. The internalization of the encapsulated material by cells was studied using a BSA-fluorescein system; the fluorescent conjugate was observable inside the cells after 20 h of incubation. The therapeutic CS-alpha interferon formulation showed a maximum of protein released in vitro at around 90 h. This system was found to be safe in a cytotoxicity assay, while biological activity experiments in vitro showed antiviral protection of cells in the presence of encapsulated porcine alpha interferon. In vivo experiments in pigs revealed a significant and sustained antiviral response through overexpression of the antiviral markers OAS2 and PKR. This proves the preservation of porcine alpha interferon biological activity, and also that a lasting response was obtained. This procedure is an effective and safe method to formulate drugs in nanoparticulate systems, representing a significant contribution to the search for more effective drug delivery strategies.


Subject(s)
Chitosan , Nanoparticles , Pharmaceutical Preparations , Animals , Antiviral Agents/pharmacology , Biological Availability , Cattle , Drug Carriers , Drug Delivery Systems , Interferon-alpha , Particle Size , Polymers , Swine
18.
Int J Environ Res Public Health ; 18(21)2021 Oct 28.
Article in English | MEDLINE | ID: covidwho-1488570

ABSTRACT

In the first quarter of 2020, Italy became one of the earliest hotspots of COVID-19 infection, and the government imposed a lockdown. During the lockdown, an online survey of 2053 adults was conducted that asked about health behaviors and about the psychological and overall impact of COVID-19. The present study is a secondary analysis of that data. We hypothesized that self-control, higher socio-economic status, existing health conditions, and fear of infection were all inversely related to actions (or intentions) that violated the lockdown (i.e., infractions). Using partial least squares structural equation modeling (PLS-SEM), we found that only the fear of infection significantly dissuaded people from violating lockdown rules. Since it is not practical or ethical to sow a fear of infection, our study indicates that enacting rules and enforcing them firmly and fairly are important tools for containing the infection. This may become more important as vaccines become more widely available and people lose their fear of infection.


Subject(s)
COVID-19 , Animals , Communicable Disease Control , Fear , Female , Humans , Italy/epidemiology , SARS-CoV-2 , Social Justice , Swine
19.
J Virol ; 95(16): e0018721, 2021 07 26.
Article in English | MEDLINE | ID: covidwho-1486048

ABSTRACT

Subversion of the host cell cycle to facilitate viral replication is a common feature of coronavirus infections. Coronavirus nucleocapsid (N) protein can modulate the host cell cycle, but the mechanistic details remain largely unknown. Here, we investigated the effects of manipulation of porcine epidemic diarrhea virus (PEDV) N protein on the cell cycle and the influence on viral replication. Results indicated that PEDV N induced Vero E6 cell cycle arrest at S-phase, which promoted viral replication (P < 0.05). S-phase arrest was dependent on the N protein nuclear localization signal S71NWHFYYLGTGPHADLRYRT90 and the interaction between N protein and p53. In the nucleus, the binding of N protein to p53 maintained consistently high-level expression of p53, which activated the p53-DREAM pathway. The key domain of the N protein interacting with p53 was revealed to be S171RGNSQNRGNNQGRGASQNRGGNN194 (NS171-N194), in which G183RG185 are core residues. NS171-N194 and G183RG185 were essential for N-induced S-phase arrest. Moreover, small molecular drugs targeting the NS171-N194 domain of the PEDV N protein were screened through molecular docking. Hyperoside could antagonize N protein-induced S-phase arrest by interfering with interaction between N protein and p53 and inhibit viral replication (P < 0.05). The above-described experiments were also validated in porcine intestinal cells, and data were in line with results in Vero E6 cells. Therefore, these results reveal the PEDV N protein interacts with p53 to activate the p53-DREAM pathway, and subsequently induces S-phase arrest to create a favorable environment for virus replication. These findings provide new insight into the PEDV-host interaction and the design of novel antiviral strategies against PEDV. IMPORTANCE Many viruses subvert the host cell cycle to create a cellular environment that promotes viral growth. PEDV, an emerging and reemerging coronavirus, has led to substantial economic loss in the global swine industry. Our study is the first to demonstrate that PEDV N-induced cell cycle arrest during the S-phase promotes viral replication. We identified a novel mechanism of PEDV N-induced S-phase arrest, where the binding of PEDV N protein to p53 maintains consistently high levels of p53 expression in the nucleus to mediate S-phase arrest by activating the p53-DREAM pathway. Furthermore, a small molecular compound, hyperoside, targeted the PEDV N protein, interfering with the interaction between the N protein and p53 and, importantly, inhibited PEDV replication by antagonizing cell cycle arrest. This study reveals a new mechanism of PEDV-host interaction and also provides a novel antiviral strategy for PEDV. These data provide a foundation for further research into coronavirus-host interactions.


Subject(s)
Antiviral Agents/pharmacology , Coronavirus Nucleocapsid Proteins/chemistry , Host-Pathogen Interactions/drug effects , Porcine epidemic diarrhea virus/drug effects , Quercetin/analogs & derivatives , Tumor Suppressor Protein p53/chemistry , Amino Acid Sequence , Animals , Antiviral Agents/chemistry , Binding Sites , Cell Line , Chlorocebus aethiops , Coronavirus Infections/drug therapy , Coronavirus Infections/genetics , Coronavirus Infections/metabolism , Coronavirus Infections/virology , Coronavirus Nucleocapsid Proteins/antagonists & inhibitors , Coronavirus Nucleocapsid Proteins/genetics , Coronavirus Nucleocapsid Proteins/metabolism , Epithelial Cells/drug effects , Epithelial Cells/virology , Gene Expression Regulation , High-Throughput Screening Assays , Host-Pathogen Interactions/genetics , Molecular Docking Simulation , Nuclear Localization Signals , Porcine epidemic diarrhea virus/genetics , Porcine epidemic diarrhea virus/metabolism , Protein Binding , Protein Conformation , Protein Interaction Domains and Motifs , Quercetin/chemistry , Quercetin/pharmacology , S Phase Cell Cycle Checkpoints/drug effects , S Phase Cell Cycle Checkpoints/genetics , Signal Transduction , Swine , Swine Diseases/drug therapy , Swine Diseases/genetics , Swine Diseases/metabolism , Swine Diseases/virology , Tumor Suppressor Protein p53/antagonists & inhibitors , Tumor Suppressor Protein p53/genetics , Tumor Suppressor Protein p53/metabolism , Vero Cells , Virus Replication/drug effects
20.
MAbs ; 13(1): 1987180, 2021.
Article in English | MEDLINE | ID: covidwho-1483313

ABSTRACT

The global health crisis and economic tolls of COVID-19 necessitate a panoply of strategies to treat SARS-CoV-2 infection. To date, few treatment options exist, although neutralizing antibodies against the spike glycoprotein have proven to be effective. Because infection is initiated at the mucosa and propagates mainly at this site throughout the course of the disease, blocking the virus at the mucosal milieu should be effective. However, administration of biologics to the mucosa presents a substantial challenge. Here, we describe bifunctional molecules combining single-domain variable regions that bind to the polymeric Ig receptor (pIgR) and to the SARS-CoV-2 spike protein via addition of the ACE2 extracellular domain (ECD). The hypothesis behind this design is that pIgR will transport the molecule from the circulation to the mucosal surface where the ACE ECD would act as a decoy receptor for the nCoV2. The bifunctional molecules bind SARS-Cov-2 spike glycoprotein in vitro and efficiently transcytose across the lung epithelium in human tissue-based analyses. Designs featuring ACE2 tethered to the C-terminus of the Fc do not induce antibody-dependent cytotoxicity against pIgR-expressing cells. These molecules thus represent a potential therapeutic modality for systemic administration of neutralizing anti-SARS-CoV-2 molecules to the mucosa.


Subject(s)
Antibodies, Viral , COVID-19/drug therapy , Receptors, Polymeric Immunoglobulin , SARS-CoV-2/immunology , Single-Chain Antibodies , Spike Glycoprotein, Coronavirus/immunology , Angiotensin-Converting Enzyme 2/genetics , Angiotensin-Converting Enzyme 2/immunology , Animals , Antibodies, Viral/genetics , Antibodies, Viral/immunology , Antibodies, Viral/pharmacology , CHO Cells , COVID-19/genetics , COVID-19/immunology , Cricetulus , Dogs , Female , Humans , Madin Darby Canine Kidney Cells , Mice , Mouth Mucosa/immunology , Protein Domains , Receptors, Polymeric Immunoglobulin/genetics , Receptors, Polymeric Immunoglobulin/immunology , Receptors, Polymeric Immunoglobulin/therapeutic use , SARS-CoV-2/genetics , Single-Chain Antibodies/genetics , Single-Chain Antibodies/immunology , Single-Chain Antibodies/pharmacokinetics , Single-Chain Antibodies/pharmacology , Spike Glycoprotein, Coronavirus/antagonists & inhibitors , Spike Glycoprotein, Coronavirus/genetics , Swine
SELECTION OF CITATIONS
SEARCH DETAIL
...