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1.
Cell ; 184(1): 64-75.e11, 2021 01 07.
Artículo en Inglés | MEDLINE | ID: mdl-33275900

RESUMEN

Global dispersal and increasing frequency of the SARS-CoV-2 spike protein variant D614G are suggestive of a selective advantage but may also be due to a random founder effect. We investigate the hypothesis for positive selection of spike D614G in the United Kingdom using more than 25,000 whole genome SARS-CoV-2 sequences. Despite the availability of a large dataset, well represented by both spike 614 variants, not all approaches showed a conclusive signal of positive selection. Population genetic analysis indicates that 614G increases in frequency relative to 614D in a manner consistent with a selective advantage. We do not find any indication that patients infected with the spike 614G variant have higher COVID-19 mortality or clinical severity, but 614G is associated with higher viral load and younger age of patients. Significant differences in growth and size of 614G phylogenetic clusters indicate a need for continued study of this variant.


Asunto(s)
Sustitución de Aminoácidos , COVID-19/transmisión , COVID-19/virología , SARS-CoV-2/genética , SARS-CoV-2/patogenicidad , Glicoproteína de la Espiga del Coronavirus/genética , Ácido Aspártico/análisis , Ácido Aspártico/genética , COVID-19/epidemiología , Genoma Viral , Glicina/análisis , Glicina/genética , Humanos , Mutación , SARS-CoV-2/crecimiento & desarrollo , Reino Unido/epidemiología , Virulencia , Secuenciación Completa del Genoma
2.
Cell ; 184(5): 1171-1187.e20, 2021 03 04.
Artículo en Inglés | MEDLINE | ID: mdl-33621484

RESUMEN

SARS-CoV-2 can mutate and evade immunity, with consequences for efficacy of emerging vaccines and antibody therapeutics. Here, we demonstrate that the immunodominant SARS-CoV-2 spike (S) receptor binding motif (RBM) is a highly variable region of S and provide epidemiological, clinical, and molecular characterization of a prevalent, sentinel RBM mutation, N439K. We demonstrate N439K S protein has enhanced binding affinity to the hACE2 receptor, and N439K viruses have similar in vitro replication fitness and cause infections with similar clinical outcomes as compared to wild type. We show the N439K mutation confers resistance against several neutralizing monoclonal antibodies, including one authorized for emergency use by the US Food and Drug Administration (FDA), and reduces the activity of some polyclonal sera from persons recovered from infection. Immune evasion mutations that maintain virulence and fitness such as N439K can emerge within SARS-CoV-2 S, highlighting the need for ongoing molecular surveillance to guide development and usage of vaccines and therapeutics.


Asunto(s)
COVID-19/inmunología , Aptitud Genética , Evasión Inmune , SARS-CoV-2/genética , Glicoproteína de la Espiga del Coronavirus/genética , Enzima Convertidora de Angiotensina 2/química , Anticuerpos Neutralizantes/genética , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , COVID-19/virología , Humanos , Mutación , Filogenia , SARS-CoV-2/química , SARS-CoV-2/patogenicidad , Glicoproteína de la Espiga del Coronavirus/química , Virulencia
3.
Nature ; 617(7961): 555-563, 2023 May.
Artículo en Inglés | MEDLINE | ID: mdl-36996873

RESUMEN

An outbreak of acute hepatitis of unknown aetiology in children was reported in Scotland1 in April 2022 and has now been identified in 35 countries2. Several recent studies have suggested an association with human adenovirus with this outbreak, a virus not commonly associated with hepatitis. Here we report a detailed case-control investigation and find an association between adeno-associated virus 2 (AAV2) infection and host genetics in disease susceptibility. Using next-generation sequencing, PCR with reverse transcription, serology and in situ hybridization, we detected recent infection with AAV2 in plasma and liver samples in 26 out of 32 (81%) cases of hepatitis compared with 5 out of 74 (7%) of samples from unaffected individuals. Furthermore, AAV2 was detected within ballooned hepatocytes alongside a prominent T cell infiltrate in liver biopsy samples. In keeping with a CD4+ T-cell-mediated immune pathology, the human leukocyte antigen (HLA) class II HLA-DRB1*04:01 allele was identified in 25 out of 27 cases (93%) compared with a background frequency of 10 out of 64 (16%; P = 5.49 × 10-12). In summary, we report an outbreak of acute paediatric hepatitis associated with AAV2 infection (most likely acquired as a co-infection with human adenovirus that is usually required as a 'helper virus' to support AAV2 replication) and disease susceptibility related to HLA class II status.


Asunto(s)
Infecciones por Adenovirus Humanos , Dependovirus , Hepatitis , Niño , Humanos , Enfermedad Aguda/epidemiología , Infecciones por Adenovirus Humanos/epidemiología , Infecciones por Adenovirus Humanos/genética , Infecciones por Adenovirus Humanos/virología , Alelos , Estudios de Casos y Controles , Linfocitos T CD4-Positivos/inmunología , Coinfección/epidemiología , Coinfección/virología , Dependovirus/aislamiento & purificación , Predisposición Genética a la Enfermedad , Virus Helper/aislamiento & purificación , Hepatitis/epidemiología , Hepatitis/genética , Hepatitis/virología , Hepatocitos/virología , Cadenas HLA-DRB1/genética , Cadenas HLA-DRB1/inmunología , Hígado/virología
4.
Nucleic Acids Res ; 52(6): 3199-3212, 2024 Apr 12.
Artículo en Inglés | MEDLINE | ID: mdl-38407436

RESUMEN

Productive infections by RNA viruses require faithful replication of the entire genome. Yet many RNA viruses also produce deletion-containing viral genomes (DelVGs), aberrant replication products with large internal deletions. DelVGs interfere with the replication of wild-type virus and their presence in patients is associated with better clinical outcomes. The DelVG RNA itself is hypothesized to confer this interfering activity. DelVGs antagonize replication by out-competing the full-length genome and triggering innate immune responses. Here, we identify an additionally inhibitory mechanism mediated by a new class of viral proteins encoded by DelVGs. We identified hundreds of cryptic viral proteins translated from DelVGs. These DelVG-encoded proteins (DPRs) include canonical viral proteins with large internal deletions, as well as proteins with novel C-termini translated from alternative reading frames. Many DPRs retain functional domains shared with their full-length counterparts, suggesting they may have activity during infection. Mechanistic studies of DPRs derived from the influenza virus protein PB2 showed that they poison replication of wild-type virus by acting as dominant-negative inhibitors of the viral polymerase. These findings reveal that DelVGs have a dual inhibitory mechanism, acting at both the RNA and protein level. They further show that DPRs have the potential to dramatically expand the functional proteomes of diverse RNA viruses.


Asunto(s)
Genoma Viral , Virus de la Influenza A , Proteoma , Proteínas Virales , Humanos , Genoma Viral/genética , Virus de la Influenza A/genética , Proteoma/genética , ARN Viral/genética , ARN Viral/metabolismo , Proteínas Virales/genética , Proteínas Virales/metabolismo , Replicación Viral/genética , Eliminación de Secuencia/genética , Animales , Perros , Línea Celular
5.
PLoS Biol ; 20(4): e3001580, 2022 04.
Artículo en Inglés | MEDLINE | ID: mdl-35439242

RESUMEN

Vaccination is a powerful tool in combating infectious diseases of humans and companion animals. In most wildlife, including reservoirs of emerging human diseases, achieving sufficient vaccine coverage to mitigate disease burdens remains logistically unattainable. Virally vectored "transmissible" vaccines that deliberately spread among hosts are a potentially transformative, but still theoretical, solution to the challenge of immunising inaccessible wildlife. Progress towards real-world application is frustrated by the absence of frameworks to guide vector selection and vaccine deployment prior to major in vitro and in vivo investments in vaccine engineering and testing. Here, we performed deep sequencing on field-collected samples of Desmodus rotundus betaherpesvirus (DrBHV), a candidate vector for a transmissible vaccine targeting vampire bat-transmitted rabies. We discovered 11 strains of DrBHV that varied in prevalence and geographic distribution across Peru. The phylogeographic structure of DrBHV strains was predictable from both host genetics and landscape topology, informing long-term DrBHV-vectored vaccine deployment strategies and identifying geographic areas for field trials where vaccine spread would be naturally contained. Multistrain infections were observed in 79% of infected bats. Resampling of marked individuals over 4 years showed within-host persistence kinetics characteristic of latency and reactivation, properties that might boost individual immunity and lead to sporadic vaccine transmission over the lifetime of the host. Further, strain acquisitions by already infected individuals implied that preexisting immunity and strain competition are unlikely to inhibit vaccine spread. Our results support the development of a transmissible vaccine targeting a major source of human and animal rabies in Latin America and show how genomics can enlighten vector selection and deployment strategies for transmissible vaccines.


Asunto(s)
Quirópteros , Rabia , Vacunas , Animales , Vectores de Enfermedades , Secuenciación de Nucleótidos de Alto Rendimiento , Rabia/epidemiología , Rabia/prevención & control , Rabia/veterinaria
6.
PLoS Genet ; 18(9): e1010406, 2022 09.
Artículo en Inglés | MEDLINE | ID: mdl-36121852

RESUMEN

Wolbachia are widespread maternally-transmitted bacteria of arthropods that often spread by manipulating their host's reproduction through cytoplasmic incompatibility (CI). Their invasive potential is currently being harnessed in field trials aiming to control mosquito-borne diseases. Wolbachia genomes commonly harbour prophage regions encoding the cif genes which confer their ability to induce CI. Recently, a plasmid-like element was discovered in wPip, a Wolbachia strain infecting Culex mosquitoes; however, it is unclear how common such extra-chromosomal elements are in Wolbachia. Here we sequenced the complete genome of wAlbA, a strain of the symbiont found in Aedes albopictus, after eliminating the co-infecting and higher density wAlbB strain that previously made sequencing of wAlbA challenging. We show that wAlbA is associated with two new plasmids and identified additional Wolbachia plasmids and related chromosomal islands in over 20% of publicly available Wolbachia genome datasets. These plasmids encode a variety of accessory genes, including several phage-like DNA packaging genes as well as genes potentially contributing to host-symbiont interactions. In particular, we recovered divergent homologues of the cif genes in both Wolbachia- and Rickettsia-associated plasmids. Our results indicate that plasmids are common in Wolbachia and raise fundamental questions around their role in symbiosis. In addition, our comparative analysis provides useful information for the future development of genetic tools to manipulate and study Wolbachia symbionts.


Asunto(s)
Aedes , Wolbachia , Aedes/genética , Animales , Plásmidos/genética , Profagos/genética , Simbiosis/genética , Wolbachia/genética
7.
Curr Issues Mol Biol ; 46(1): 485-497, 2024 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-38248333

RESUMEN

Oral squamous cell carcinoma (OSCC) is a common and highly aggressive dog tumor known for its local invasiveness and metastatic potential. Understanding the molecular mechanisms driving the development and progression of OSCC is crucial for improving diagnostic and therapeutic strategies. Additionally, spontaneous oral squamous cell carcinomas in dogs are an excellent model for studying human counterparts. In this study, we aimed to investigate the significance of two key molecular components, Cox-2 and EGFR, in canine OSCC. We examined 34 tumor sections from various dog breeds to assess the immunoexpression of Cox-2 and EGFR. Our findings revealed that Cox-2 was highly expressed in 70.6% of cases, while EGFR overexpression was observed in 44.1%. Cox-2 overexpression showed association with histological grade of malignancy (HGM) (p = 0.006) and EGFR with vascular invasion (p = 0.006). COX-2 and EGFR concurrent expression was associated with HGM (p = 0.002), as well as with the presence of vascular invasion (p = 0.002). These data suggest that Cox-2 and EGFR could be promising biomarkers and potential therapeutic targets, opening avenues for developing novel treatment strategies for dogs affected by OSCC. Further studies are warranted to delve deeper into these findings and translate them into clinical practice.

8.
Curr Issues Mol Biol ; 46(5): 4951-4967, 2024 May 19.
Artículo en Inglés | MEDLINE | ID: mdl-38785565

RESUMEN

Squamous cell carcinoma (SCC) stands as the second most prevalent skin cancer in dogs, primarily attributed to UV radiation exposure. Affected areas typically include regions with sparse hair and pale or depigmented skin. The significance of spontaneous canine cutaneous SCC as a model for its human counterpart is underscored by its resemblance. This study assesses the expression of key markers-Epidermal Growth Factor Receptor (EGFR), Cyclooxygenase-2 (Cox-2), and Ki-67-in canine cutaneous SCC. Our objective is to investigate the association between their expression levels and classical clinicopathological parameters, unraveling the intricate relationships among these molecular markers. In our retrospective analysis of 37 cases, EGFR overexpression manifested in 43.2% of cases, while Cox-2 exhibited overexpression in 97.3%. The EGFR, Cox-2 overexpression, and Ki-67 proliferation indices, estimated through immunohistochemistry, displayed a significant association with the histological grade, but only EGFR labeling is associated with the presence of lymphovascular emboli. The Ki-67 labeling index expression exhibited an association with EGFR and Cox-2. These findings propose that EGFR, Cox-2, and Ki-67 hold promise as valuable markers in canine SCC. EGFR, Cox-2, and Ki-67 may serve as indicators of disease progression, offering insights into the malignancy of a lesion. The implications extend to the potential therapeutic targeting of EGFR and Cox-2 in managing canine SCC. Further exploration of these insights is warranted due to their translational relevance and the development of targeted interventions in the context of canine SCC.

9.
PLoS Pathog ; 18(11): e1010973, 2022 11.
Artículo en Inglés | MEDLINE | ID: mdl-36399512

RESUMEN

HIV-1 transmission via sexual exposure is an inefficient process. When transmission does occur, newly infected individuals are colonized by the descendants of either a single virion or a very small number of establishing virions. These transmitted founder (TF) viruses are more interferon (IFN)-resistant than chronic control (CC) viruses present 6 months after transmission. To identify the specific molecular defences that make CC viruses more susceptible to the IFN-induced 'antiviral state', we established a single pair of fluorescent TF and CC viruses and used arrayed interferon-stimulated gene (ISG) expression screening to identify candidate antiviral effectors. However, we observed a relatively uniform ISG resistance of transmitted HIV-1, and this directed us to investigate possible underlying mechanisms. Simple simulations, where we varied a single parameter, illustrated that reduced growth rate could possibly underly apparent interferon sensitivity. To examine this possibility, we closely monitored in vitro propagation of a model TF/CC pair (closely matched in replicative fitness) over a targeted range of IFN concentrations. Fitting standard four-parameter logistic growth models, in which experimental variables were regressed against growth rate and carrying capacity, to our in vitro growth curves, further highlighted that small differences in replicative growth rates could recapitulate our in vitro observations. We reasoned that if growth rate underlies apparent interferon resistance, transmitted HIV-1 would be similarly resistant to any growth rate inhibitor. Accordingly, we show that two transmitted founder HIV-1 viruses are relatively resistant to antiretroviral drugs, while their matched chronic control viruses were more sensitive. We propose that, when present, the apparent IFN resistance of transmitted HIV-1 could possibly be explained by enhanced replicative fitness, as opposed to specific resistance to individual IFN-induced defences. However, further work is required to establish how generalisable this mechanism of relative IFN resistance might be.


Asunto(s)
Dermatitis , Seropositividad para VIH , VIH-1 , Humanos , Interferones/farmacología , Antivirales , Replicación del ADN
10.
PLoS Biol ; 19(12): e3001065, 2021 12.
Artículo en Inglés | MEDLINE | ID: mdl-34932557

RESUMEN

The pandemic spread of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the etiological agent of Coronavirus Disease 2019 (COVID-19), represents an ongoing international health crisis. A key symptom of SARS-CoV-2 infection is the onset of fever, with a hyperthermic temperature range of 38 to 41°C. Fever is an evolutionarily conserved host response to microbial infection that can influence the outcome of viral pathogenicity and regulation of host innate and adaptive immune responses. However, it remains to be determined what effect elevated temperature has on SARS-CoV-2 replication. Utilizing a three-dimensional (3D) air-liquid interface (ALI) model that closely mimics the natural tissue physiology of SARS-CoV-2 infection in the respiratory airway, we identify tissue temperature to play an important role in the regulation of SARS-CoV-2 infection. Respiratory tissue incubated at 40°C remained permissive to SARS-CoV-2 entry but refractory to viral transcription, leading to significantly reduced levels of viral RNA replication and apical shedding of infectious virus. We identify tissue temperature to play an important role in the differential regulation of epithelial host responses to SARS-CoV-2 infection that impact upon multiple pathways, including intracellular immune regulation, without disruption to general transcription or epithelium integrity. We present the first evidence that febrile temperatures associated with COVID-19 inhibit SARS-CoV-2 replication in respiratory epithelia. Our data identify an important role for tissue temperature in the epithelial restriction of SARS-CoV-2 independently of canonical interferon (IFN)-mediated antiviral immune defenses.


Asunto(s)
Células Epiteliales/inmunología , Calor , Inmunidad Innata/inmunología , Interferones/inmunología , Mucosa Respiratoria/inmunología , SARS-CoV-2/inmunología , Replicación Viral/inmunología , Adolescente , Animales , COVID-19/genética , COVID-19/inmunología , COVID-19/virología , Chlorocebus aethiops , Células Epiteliales/metabolismo , Células Epiteliales/virología , Femenino , Perfilación de la Expresión Génica/métodos , Interacciones Huésped-Patógeno/genética , Interacciones Huésped-Patógeno/inmunología , Humanos , Inmunidad Innata/genética , Interferones/genética , Interferones/metabolismo , Masculino , Persona de Mediana Edad , Modelos Biológicos , RNA-Seq/métodos , Mucosa Respiratoria/metabolismo , Mucosa Respiratoria/virología , SARS-CoV-2/genética , SARS-CoV-2/fisiología , Técnicas de Cultivo de Tejidos , Células Vero , Replicación Viral/genética , Replicación Viral/fisiología
11.
PLoS Biol ; 19(2): e3001091, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33630831

RESUMEN

The recent emergence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the underlying cause of Coronavirus Disease 2019 (COVID-19), has led to a worldwide pandemic causing substantial morbidity, mortality, and economic devastation. In response, many laboratories have redirected attention to SARS-CoV-2, meaning there is an urgent need for tools that can be used in laboratories unaccustomed to working with coronaviruses. Here we report a range of tools for SARS-CoV-2 research. First, we describe a facile single plasmid SARS-CoV-2 reverse genetics system that is simple to genetically manipulate and can be used to rescue infectious virus through transient transfection (without in vitro transcription or additional expression plasmids). The rescue system is accompanied by our panel of SARS-CoV-2 antibodies (against nearly every viral protein), SARS-CoV-2 clinical isolates, and SARS-CoV-2 permissive cell lines, which are all openly available to the scientific community. Using these tools, we demonstrate here that the controversial ORF10 protein is expressed in infected cells. Furthermore, we show that the promising repurposed antiviral activity of apilimod is dependent on TMPRSS2 expression. Altogether, our SARS-CoV-2 toolkit, which can be directly accessed via our website at https://mrcppu-covid.bio/, constitutes a resource with considerable potential to advance COVID-19 vaccine design, drug testing, and discovery science.


Asunto(s)
Vacunas contra la COVID-19 , COVID-19/diagnóstico , COVID-19/virología , Genética Inversa , SARS-CoV-2/genética , Células A549 , Enzima Convertidora de Angiotensina 2/metabolismo , Animales , Chlorocebus aethiops , Codón , Humanos , Hidrazonas/farmacología , Ratones , Morfolinas/farmacología , Sistemas de Lectura Abierta , Plásmidos/genética , Pirimidinas/farmacología , Serina Endopeptidasas/metabolismo , Células Vero , Proteínas Virales/metabolismo
12.
J Neuroeng Rehabil ; 21(1): 53, 2024 Apr 10.
Artículo en Inglés | MEDLINE | ID: mdl-38600582

RESUMEN

A systematic review was conducted to determine the trends in devices and parameters used for brain photobiomodulation (PBM). The revised studies included clinical and cadaveric approaches, in which light stimuli were applied to the head and/or neck. PubMed, Scopus, Web of Science and Google Scholar databases were used for the systematic search. A total of 2133 records were screened, from which 97 were included in this review. The parameters that were extracted and analysed in each article were the device design, actuation area, actuation site, wavelength, mode of operation, power density, energy density, power output, energy per session and treatment time. To organize device information, 11 categories of devices were defined, according to their characteristics. The most used category of devices was laser handpieces, which relate to 21% of all devices, while 28% of the devices were not described. Studies for cognitive function and physiological characterisation are the most well defined ones and with more tangible results. There is a lack of consistency when reporting PBM studies, with several articles under defining the stimulation protocol, and a wide variety of parameters used for the same health conditions (e.g., Alzheimer's or Parkinson's disease) resulting in positive outcomes. Standardization for the report of these studies is warranted, as well as sham-controlled comparative studies to determine which parameters have the greatest effect on PBM treatments for different neurological conditions.


Asunto(s)
Terapia por Luz de Baja Intensidad , Humanos , Terapia por Luz de Baja Intensidad/métodos , Encéfalo , Cognición , Rayos Láser
13.
Int J Mol Sci ; 25(11)2024 May 24.
Artículo en Inglés | MEDLINE | ID: mdl-38891904

RESUMEN

Tooth loss during the lifetime of an individual is common. A strategy to treat partial or complete edentulous patients is the placement of dental implants. However, dental implants are subject to bacterial colonization and biofilm formation, which cause an infection named peri-implantitis. The existing long-term treatments for peri-implantitis are generally inefficient. Thus, an electrical circuit was produced with zirconia (Zr) samples using a hot-pressing technique to impregnate silver (Ag) through channels and holes to create a path by LASER texturing. The obtained specimens were characterized according to vitro cytotoxicity, to ensure ZrAg non-toxicity. Furthermore, samples were inoculated with Staphylococcus aureus using 6.5 mA of alternating current (AC). The current was delivered using a potentiostat and the influence on the bacterial concentration was assessed. Using AC, the specimens displayed no bacterial adhesion (Log 7 reduction). The in vitro results presented in this study suggest that this kind of treatment can be an alternative and promising strategy to treat and overcome bacterial adhesion around dental implants that can evolve to biofilm.


Asunto(s)
Adhesión Bacteriana , Biopelículas , Implantes Dentales , Staphylococcus aureus , Circonio , Implantes Dentales/microbiología , Circonio/química , Biopelículas/crecimiento & desarrollo , Biopelículas/efectos de los fármacos , Humanos , Estimulación Eléctrica/métodos , Propiedades de Superficie , Periimplantitis/microbiología , Periimplantitis/terapia , Plata/química , Plata/farmacología
14.
J Cancer Educ ; 39(3): 288-296, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38421568

RESUMEN

Leadership plays a key role in cancer education (CE) and the success of its practices. Leaders in CE must effectively use their leadership skills to be able to communicate, collaborate, and educate their team members. There is a lack of formalized and standardized curriculums for institutions in developing leadership programs, including what themes to focus on in CE. In this article, the authors describe key pillars of leadership in CE that have presented themselves throughout their experience and within the literature. A search was conducted using the Ovid MEDLINE® database and articles were reviewed for eligibility. In this review, thirty articles were selected for their relevance to CE. With this literature search and the authors' reflections, four pillars of leadership in CE were identified: (1) leadership development, (2) collaboration, (3) diversity and equity, and (4) implementation. Within these themes, key areas of importance were discussed further, and barriers to CE leadership were identified. By reflecting upon pillars of leadership in CE, this article may be helpful for developing future leadership programs within CE. It is vital that initiatives continue to be held and barriers are addressed to increase leadership effectiveness within CE.


Asunto(s)
Liderazgo , Neoplasias , Humanos , Neoplasias/prevención & control , Curriculum , Conducta Cooperativa
15.
J Gen Virol ; 104(8)2023 08.
Artículo en Inglés | MEDLINE | ID: mdl-37643006

RESUMEN

Distinct cytomegaloviruses (CMVs) are widely distributed across their mammalian hosts in a highly host species-restricted pattern. To date, evidence demonstrating this has been limited largely to PCR-based approaches targeting small, conserved genomic regions, and only a few complete genomes of isolated viruses representing distinct CMV species have been sequenced. We have now combined direct isolation of infectious viruses from tissues with complete genome sequencing to provide a view of CMV diversity in a wild animal population. We targeted Natal multimammate mice (Mastomys natalensis), which are common in sub-Saharan Africa, are known to carry a variety of zoonotic pathogens, and are regarded as the primary source of Lassa virus (LASV) spillover into humans. Using transformed epithelial cells prepared from M. natalensis kidneys, we isolated CMVs from the salivary gland tissue of 14 of 37 (36 %) animals from a field study site in Mali. Genome sequencing showed that these primary isolates represent three different M. natalensis CMVs (MnatCMVs: MnatCMV1, MnatCMV2 and MnatCMV3), with some animals carrying multiple MnatCMVs or multiple strains of a single MnatCMV presumably as a result of coinfection or superinfection. Including primary isolates and plaque-purified isolates, we sequenced and annotated the genomes of two MnatCMV1 strains (derived from sequencing 14 viruses), six MnatCMV2 strains (25 viruses) and ten MnatCMV3 strains (21 viruses), totalling 18 MnatCMV strains isolated as 60 infectious viruses. Phylogenetic analysis showed that these MnatCMVs group with other murid viruses in the genus Muromegalovirus (subfamily Betaherpesvirinae, family Orthoherpesviridae), and that MnatCMV1 and MnatCMV2 are more closely related to each other than to MnatCMV3. The availability of MnatCMV isolates and the characterization of their genomes will serve as the prelude to the generation of a MnatCMV-based vaccine to target LASV in the M. natalensis reservoir.


Asunto(s)
Infecciones por Citomegalovirus , Citomegalovirus , Animales , Humanos , Ratones , Filogenia , Secuencia de Bases , Murinae
16.
PLoS Pathog ; 17(9): e1009929, 2021 09.
Artículo en Inglés | MEDLINE | ID: mdl-34534263

RESUMEN

Remdesivir (RDV), a broadly acting nucleoside analogue, is the only FDA approved small molecule antiviral for the treatment of COVID-19 patients. To date, there are no reports identifying SARS-CoV-2 RDV resistance in patients, animal models or in vitro. Here, we selected drug-resistant viral populations by serially passaging SARS-CoV-2 in vitro in the presence of RDV. Using high throughput sequencing, we identified a single mutation in RNA-dependent RNA polymerase (NSP12) at a residue conserved among all coronaviruses in two independently evolved populations displaying decreased RDV sensitivity. Introduction of the NSP12 E802D mutation into our SARS-CoV-2 reverse genetics backbone confirmed its role in decreasing RDV sensitivity in vitro. Substitution of E802 did not affect viral replication or activity of an alternate nucleoside analogue (EIDD2801) but did affect virus fitness in a competition assay. Analysis of the globally circulating SARS-CoV-2 variants (>800,000 sequences) showed no evidence of widespread transmission of RDV-resistant mutants. Surprisingly, we observed an excess of substitutions in spike at corresponding sites identified in the emerging SARS-CoV-2 variants of concern (i.e., H69, E484, N501, H655) indicating that they can arise in vitro in the absence of immune selection. The identification and characterisation of a drug resistant signature within the SARS-CoV-2 genome has implications for clinical management and virus surveillance.


Asunto(s)
Adenosina Monofosfato/análogos & derivados , Alanina/análogos & derivados , Antivirales/farmacología , Tratamiento Farmacológico de COVID-19 , ARN Polimerasa Dependiente de ARN de Coronavirus/genética , Farmacorresistencia Microbiana/genética , SARS-CoV-2/efectos de los fármacos , Adenosina Monofosfato/farmacología , Alanina/farmacología , Animales , Evolución Biológica , Chlorocebus aethiops , Humanos , Mutación , SARS-CoV-2/genética , Glicoproteína de la Espiga del Coronavirus/metabolismo , Células Vero
17.
Rapid Commun Mass Spectrom ; 37(1): e9418, 2023 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-36261319

RESUMEN

RATIONALE: 2,4,6-Tris(4-formylphenyl)boroxine (TFPB) is a substituted boroxine containing a benzaldehyde molecule bonded to each boron atom. Boroxine cages are an emerging class of functional nanostructures used in host-guest chemistry, and benzaldehyde is a potential radiosensitizer. Reactions initiated by low-energy electrons with such complexes may dictate and bring new fundamental knowledge for biomedical and pharmaceutical applications. METHODS: The electron ionization properties of TFPB are investigated using a gas-phase electron-molecule crossed beam apparatus coupled with a reflectron time-of-flight mass spectrometer in an orthogonal geometry. Ionization and threshold energies are experimentally determined by mass spectra acquisition as a function of the electron energy. RESULTS: The abundance of the molecular precursor cation in the mass spectrum at 70 eV is significantly lower than that of the most abundant fragment C7 H5 O+ . Twenty-nine cationic fragments with relative intensities >2% are detected and identified. The appearance energies of six fragment cations are reported, and the experimental first ionization potential is found at 9.46 ± 0.11  eV. Moreover, eight double cations are identified. The present results are supported by quantum chemical calculations based on bound state techniques, electron ionization models and thermodynamic thresholds. CONCLUSIONS: According to these results, the TPFB properties may combine the potential radiosensitizer effect of benzaldehyde with the stability of the boroxine ring.


Asunto(s)
Benzaldehídos , Electrones , Cationes , Preparaciones Farmacéuticas
18.
Phys Chem Chem Phys ; 25(37): 25361-25367, 2023 Sep 27.
Artículo en Inglés | MEDLINE | ID: mdl-37703052

RESUMEN

Trichloroanisole (TCA) is one of the most significant contaminants in cork stoppers. The presence of TCA leads to an unpleasant odor known as "cork taint", resulting in high economic losses for the cork and wine industries. Hence, the detection, quantification, and characterization of TCA are essential to address this concern. The present study investigates the electron-driven fragmentation pathways of TCA through electron ionization mass spectrometry as a function of electron energy (0-100 eV), and the results are supported by theoretical characterization of ionization potentials, dissociation thresholds, and electron ionization cross sections. The appearance energies of ten cations were measured, including the first experimental evaluation of the molecule's ionization energy at 8.8 ± 0.3 eV, in excellent agreement with the calculations (8.83 eV). For lower energies, around 20 eV, the parent cation accounted for more than 60% of the total ion signal, followed by its demethylated fragment. Taken together, these ion signals could be used as fingerprints of TCA in industrial quality control by low-energy electron ionization mass spectrometry. Fifty other fragments have been identified at higher electron energies, revealing the very rich fragmentation pattern of TCA.

19.
Appetite ; 180: 106355, 2023 01 01.
Artículo en Inglés | MEDLINE | ID: mdl-36341861

RESUMEN

The global food consumption scenario and the influence of a country's image on consumer decision-making motivated this research. The global edible insect market is growing, and cultural issues, disgust, and low perception of quality are barriers to consumption. Through an online experiment (n = 194) Brazilian consumers demonstrate that the use of the country-of-origin label (COOL) can result in greater intention and quality expectation regarding a cricket flour. Consumers showed a preference for the flour produced by a positive image country label. Both intention and quality expectation were higher for the cricket flour produced in the United States. Furthermore, the mediation model indicates that quality expectation mediated the effect of COOL on the intention to consume. The results show that even though consumers have created a positive expectation concerning the product, a higher level of neophobia diminished their intention to consume the product and this relates to the aversion and disgust that can exist towards an edible-insect product. The neophobia level can weaken the effect of COOL on intention mediated by quality expectation. The study aimed to contribute to a better understanding of the consumer's perception of an insect-based food and provide insights into variables related to the intention to consume by verifying their contribution to the consumer's behavior.


Asunto(s)
Insectos , Humanos , Animales , Brasil
20.
Clin Oral Investig ; 27(7): 3331-3345, 2023 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-37069409

RESUMEN

OBJECTIVE: The purpose of this study was to perform an integrative review on laser texturing the inner surface of lithium disilicate-reinforced glass ceramic or zirconia to increase their bond strength to resin-matrix cements. MATERIALS AND METHOD: A bibliographic review was performed on PubMed using the following search terms: "zirconia" OR "lithium disilicate" AND "laser" AND "surface" OR "roughness" AND "bond strength" AND "luting agent" OR "resin cement." Studies published in English language until March 15, 2023, were selected regarding the purpose of this study. RESULTS: A total of fifty-six studies were identified althoug thirteen studies were selected. The findings revealed that zirconia surfaces were significantly modified after laser irradiation resulting in macro-scale aligned retentive regions with depth values ranging from 50 to 120 µm. Average roughness values of laser-textured zirconia by Er,Cr:YSGG laser (~ 0.83 µm) were quite similar when compared to grit-blasted zirconia surfaces (~ 0.9 µm) although roughness increased up to 2.4 µm depending on the laser type and parameters. Lithium disilicate-reinforced glass ceramics textured with Er:YAG revealed an average roughness of around 3.5 µm while surfaces textured using Nd:YAG laser revealed an average roughness of 2.69 µm; that was quite similar to the roughness values recorded for etched surfaces (2.64 µm). The shear bond strength (SBS) values of zirconia surfaces textured on Nd:YVO4 laser irradiation were slightly higher (~ 33.5 MPa) than those recorded for grit-blasted zirconia surfaces (28 MPa). Laser-textured zirconia surfaces on CO2 laser revealed higher SBS values (18.1 ±0.8 MPa) than those (9.1 ± 0.56 MPa) recorded for untreated zirconia surfaces. On lithium disilicate-reinforced glass ceramics, higher SBS values to resin-matrix cements were recorded for specimens textured with a combination of fractional CO2 laser irradiation and HF acid etching (~ 22-24 MPa) when compared with grit-blasted specimens (12.2 MPa). Another study revealed SBS values at around 27.5 MPa for Er:YAG-textured lithium disilicate-reinforced glass ceramics to resin-matrix cements. CONCLUSIONS: The laser irradiation at high power increases the roughness of the inner surface of lithium disilicate-reinforced glass ceramic or zirconia leading to an enhanced bond strength to resin-matrix cements. Thus, the laser type and irradiation parameters can be adjusted to enhance the macro- and micro-scale retention of zirconia and glass ceramic surfaces to resin-matrix cements. CLINICAL RELEVANCE: Alternative methods for surface modification of lithium disilicate-reinforced glass ceramic and zirconia surfaces have been assessed to provide proper morphological aspects for enhanced adhesion to resin-matrix cements. An increase in the bond strength of glass ceramics or zirconia to resin-matrix cements can improve the long-term performance of cemented prosthetic structures in the oral cavity.


Asunto(s)
Recubrimiento Dental Adhesivo , Láseres de Estado Sólido , Cementos de Resina/química , Litio , Dióxido de Carbono , Propiedades de Superficie , Cerámica/química , Porcelana Dental/química , Resistencia al Corte , Ensayo de Materiales , Circonio/química
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