RESUMEN
The recent reinforcement of CoV surveillance in animals fuelled by the COVID-19 pandemic provided increasing evidence that mammals other than bats might hide further diversity and play critical roles in human infectious diseases. This work describes the results of a two-year survey carried out in Italy with the double objective of uncovering CoV diversity associated with wildlife and of excluding the establishment of a reservoir for SARS-CoV-2 in particularly susceptible or exposed species. The survey targeted hosts from five different orders and was harmonised across the country in terms of sample size, target tissues, and molecular test. Results showed the circulation of 8 CoV species in 13 hosts out of the 42 screened. Coronaviruses were either typical of the host species/genus or normally associated with their domestic counterpart. Two novel viruses likely belonging to a novel CoV genus were found in mustelids. All samples were negative for SARS-CoV-2, with minimum detectable prevalence ranging between 0.49% and 4.78% in the 13 species reaching our threshold sample size of 59 individuals. Considering that within-species transmission in white-tailed deer resulted in raising the prevalence from 5% to 81% within a few months, this result would exclude a sustained cycle after spillback in the tested species.
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COVID-19 , Quirópteros , Ciervos , Salud Única , Animales , Humanos , Animales Salvajes , COVID-19/epidemiología , COVID-19/veterinaria , SARS-CoV-2 , PandemiasRESUMEN
All genes of avian influenza A viruses are phylogenetically distinguished into two large clades, namely the American and Eurasian clade. Reassortments among the gene segments of influenza viruses belonging to the two distinct clades are rare events and have never been described in poultry in Europe and Asia before. This study presents the genetic characterization of two influenza viruses isolated from domestic mallards in Italy in 2004 and 2005. Phylogenetic analysis of the entire genome showed that these viruses contain mixed gene segments belonging to the American and Eurasian clades.
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Virus de la Influenza A/genética , Virus Reordenados/genética , Recombinación Genética , Animales , Anseriformes , Asia , Europa (Continente) , Virus de la Influenza A/clasificación , Virus de la Influenza A/aislamiento & purificación , Gripe Aviar/virología , Italia , Datos de Secuencia Molecular , Filogenia , Virus Reordenados/clasificación , Virus Reordenados/aislamiento & purificaciónRESUMEN
The genus Pestivirus comprises globally distributed members of the family Flaviviridae, which cause severe losses in livestock. The most common species of the genus are bovine viral diarrhoea virus type 1 (BVDV-1) and type 2 (BVDV-2), classical swine fever virus (CSFV) and border disease virus (BDV). Recently, a novel ovine pestivirus was repeatedly detected in aborted lamb foetuses on a farm located in the Brescia Province (Italy). Complete genome characterization of this isolate showed that it was highly divergent from known pestivirus species and that it was genetically closely related to CSFV. The aim of this study was to determine the serological relatedness between the identified novel pestivirus and BVDV, BDV and CSFV selected strains for which homologous serum was available, by antigenic characterization performed using cross-neutralization assays. The serological relatedness was expressed as the coefficient of antigenic similarity (R). Both field and specific antisera raised against the ovine pestivirus neutralized the CSFV reference strain Diepholz with titres significantly higher than those specific for the BDV and BVDV strains. Furthermore, the calculated R values clearly indicated that the novel ovine pestivirus is antigenically more related to CSFV than to ruminant pestiviruses, in agreement with the results of the genomic analysis. This would have severe consequences on CSFV serology in the event of a switch to porcine hosts with implications for CSFV surveillance and porcine health management.
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Peste Porcina Clásica/virología , Pestivirus/genética , Enfermedades de las Ovejas/virología , Animales , Peste Porcina Clásica/epidemiología , Italia/epidemiología , Pestivirus/clasificación , Rumiantes/virología , Ovinos , Enfermedades de las Ovejas/epidemiología , PorcinosRESUMEN
Aujeszky's disease in cattle is caused by Suid herpes virus 1. The natural infection has been reported worldwide in bovine species and it is related to direct and indirect contact with infected pigs, which represent the main reservoir of the virus. Here, it is reported the first documented outbreak of Aujeszky's disease in cattle in Sicily (Italy). Severe itching and nonspecific neurological symptoms were the main reported clinical signs. No characteristic gross and histological features were reported other than cutaneous lesions caused by excessive pruritus and hyperaemia, haemorrhages and inflammation in the central nervous system. Diagnosis was confirmed by real time PCR and immunohistochemistry on the nervous tissue. The route of infection remained unknown, but serological data observed in pigs living in close cohabitation with cattle revealed a circulation of a wild strain of the virus in the area. This study contributes to a better knowledge of this disease in a non-conventional host and suggests the need to increase the prophylaxis control plans in specific breeding contexts.
RESUMEN
PURPOSE: Although doxorubicin is widely used to treat cancer, severe side effects limit its clinical use. Combination of standard chemotherapy with natural products can increase the efficacy and attenuate the side effects of current therapies. Here we studied the anticancer effects of a combined regimen comprising doxorubicin and [10]-gingerol against triple-negative breast cancer, which does not respond to hormonal or targeted therapies. METHODS: Cytotoxicity was evaluated by MTT assay, cell cycle progression and apoptosis were analyzed by flow cytometry and signaling pathways were analyzed by Western blotting in human and murine triple negative breast cancer cell systems. The anticancer/antimetastatic and toxic effects of the combined regimen was evaluated using syngeneic and xenograft orthotopic models. RESULTS: The combination of doxorubicin and [10]-gingerol significantly increased the number of apoptotic cells, compared to each compound alone. In 4T1Br4 cells, the combined regimen was the only condition able to increase the levels of active caspase 3 and γH2AX and to decrease the level of Cdk-6 cyclin. In vivo, doxorubicin (3 mg/Kg, D3) and [10]-gingerol (10 mg/Kg, G10) resulted in a significant reduction in the volume of primary tumors and a decrease in the number of circulating tumor cells (CTCs). Interestingly, only the combined regimen led to decreased tumor burdens to distant organs (i.e., metastasis) and reduced chemotherapy-induced weight loss and hepatotoxicity in tumor-bearing animals. Likewise, in a xenograft model, only the combined regimen was effective in significantly reducing the primary tumor volume and the prevalence of CTCs. CONCLUSIONS: Our data indicate that [10]-gingerol has potential to be used as a neoadjuvant or in combined therapy with doxorubicin, to improve its anticancer activity.
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Antineoplásicos/uso terapéutico , Catecoles/farmacología , Doxorrubicina/efectos adversos , Doxorrubicina/uso terapéutico , Alcoholes Grasos/farmacología , Neoplasias de la Mama Triple Negativas/tratamiento farmacológico , Animales , Antineoplásicos/efectos adversos , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Puntos de Control del Ciclo Celular/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Doxorrubicina/farmacología , Sinergismo Farmacológico , Humanos , Concentración 50 Inhibidora , Ratones Endogámicos BALB C , Metástasis de la Neoplasia , Ensayo de Tumor de Célula Madre , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Although the eradication of Pseudorabies virus (PrV) in domestic pigs is ongoing, the circulation of this virus in wild boars remains a threat in the currently unprotected, 'low prevalence', pig population. In this study, we reported PrV prevalence data and the influence of possible risk factors in 2 North-West Italian wild boar populations (free and enclosed) with the goal of supporting the implementation of a risk-based AD surveillance system. Sera from 1,425 wild boars were collected between 2011 and 2015 and tested by ELISA for the presence of PrV antibodies; the overall raw seroprevalence was 30.39% (433/1,425; CI 95% 28.01-32.85%). A significant difference was however observed between the prevalence rates of the free range population (9.98%; 90/902; CI 95%; 8.10-12.12%) and the enclosed population of La Mandria park (Piedmont, Italy) (65.58%; 343/523; CI 95%; 61.51-69.65%). In both populations a significantly higher number of adults and females were found positive to PrV ELISA. Specific territorial data on PrV circulation in wild boars should be acquired from other regions for guiding risk-based measures in order to reduce the threat of AD re-infection in a more cost-effective manner.
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Monitoreo Epidemiológico/veterinaria , Herpesvirus Suido 1/aislamiento & purificación , Seudorrabia/epidemiología , Enfermedades de los Porcinos/epidemiología , Animales , Italia/epidemiología , Prevalencia , Seudorrabia/virología , Factores de Riesgo , Estudios Seroepidemiológicos , Sus scrofa , Porcinos , Enfermedades de los Porcinos/virologíaRESUMEN
BACKGROUND AND OBJECTIVES: Radiofrequency ablation of genicular nerves has proved to be successful in relieving pain and incapacity caused by osteoarthritis of the knee. However, long-term efficacy of such a treatment remains to be assessed. The current study aimed to reproduce radiofrequency neurotomy of genicular nerves to manage gonarthrosis pain and disability and establish therapeutic response until 1 year after intervention. METHODS: This single-center, prospective, observational, noncontrolled, longitudinal study included patients with grade 3 to 4 gonarthrosis suffering from intractable knee pain, scoring 5 or more on the visual analog scale (VAS) during >6 months. Therapy was based on ultrasound guided radiofrequency neurotomy of the superior medial, superior lateral and inferior medial genicular nerves. Visual analog scale and Western Ontario and McMaster Universities Osteoarthritis scores were assessed before therapy and at 1, 6, and 12 months following treatment. RESULTS: Radiofrequency neurotomy of genicular nerves significantly reduced perceived pain (VAS) and disability (Western Ontario and McMaster Universities Osteoarthritis) in the majority of participants, without untoward events. The proportion of participants with improvement of 50% or greater in pretreatment VAS scores at 1, 6, and 12 months following intervention were 22/25 (88%), 16/25 (64%) and 8/25 (32%), respectively. CONCLUSIONS: Ultrasound-guided radiofrequency neurotomy of genicular nerves alleviates intractable pain and disability in the majority of patients with advanced osteoarthritis of the knee. Such a treatment is safe and minimally invasive and can be performed in an outpatient setting. The beneficial effect of treatment started to decline after 6 months, but even 1 year after the intervention, 32% of patients reported 50% improvement or greater in pretreatment VAS scores.
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Analgesia/tendencias , Ablación por Catéter/tendencias , Articulación de la Rodilla/inervación , Osteoartritis de la Rodilla/cirugía , Dimensión del Dolor/tendencias , Recuperación de la Función , Anciano , Anciano de 80 o más Años , Analgesia/métodos , Ablación por Catéter/métodos , Femenino , Estudios de Seguimiento , Humanos , Articulación de la Rodilla/diagnóstico por imagen , Articulación de la Rodilla/fisiología , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Osteoartritis de la Rodilla/diagnóstico por imagen , Dimensión del Dolor/métodos , Estudios Prospectivos , Recuperación de la Función/fisiología , Resultado del TratamientoRESUMEN
Sixteen haemagglutinin (HA) subtypes of avian influenza viruses (AIV) have been described to date. Rapid subtype identification of any AIV is of major interest because of the possible serious consequences for the poultry industry and even public health. Molecular techniques currently allow immediate accurate subtype characterisation prior to virus isolation. In this study, a set of fourteen specific real-time RT-PCR methods were developed and evaluated for AIV HA subtyping (H1-H4, H6-H8, H10-H16), H5 and H9 being excluded on the basis of the current validity of the European Union (EU) recommended specific assays. Specific primers and probes sets for each HA-subtype were designed to hybridise the largest isolates range within each single subtype, considering the Eurasian lineage as a major target. The robustness and general application of the 14 HA-subtype methods were verified by the analysis of 110 AIV isolates belonging to all 16 HA-subtypes, performed in different laboratories. The developed real-time RT-PCR assays proved to be highly specific and revealed suitable sensitivity, allowing direct HA-subtyping of clinical material. In summary, this study provides for the first time a panel of molecular tests using specific hydrolysis probes for rapid and complete AIV HA-subtype identification.
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Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Virus de la Influenza A/clasificación , Virus de la Influenza A/aislamiento & purificación , Gripe Aviar/virología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Virología/métodos , Animales , Cartilla de ADN/genética , Unión Europea , Virus de la Influenza A/genética , Sondas de Oligonucleótidos/genética , Aves de Corral , Sensibilidad y EspecificidadRESUMEN
This study describes an optimization of [6]-, [8]- and [10]-gingerol isolation and purification in semi-preparative HPLC scale and their anti-proliferative activity. The gingerols purification was carried out in HPLC system using a Luna-C18 and the best mobile phase evaluated was MeOH/H2O (75:25, v/v). This new methodology for the gingerols isolation was very effective, since considerable amounts (in the range of milligrams) with a good purity degree (â¼98%) were achieved in 30 min of chromatographic run. [6]-, [8]- and [10]-Gingerol purified by this methodology inhibited the proliferation of MDA-MB-231 tumor cell line with IC50 of 666.2±134.6 µM, 135.6±22.6 µM and 12.1±0.3 µM, respectively. These substances also inhibited human fibroblasts (HF) cell proliferation, however in concentrations starting from 500 µM. In conclusion, our results demonstrate an optimization of gingerols isolation and their specific anti-proliferative activities against tumor cells, suggesting their use as important models for drug design in an attempt to develop new compounds with fewer side effects when compared to conventional chemotherapy.
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Catecoles/aislamiento & purificación , Catecoles/farmacología , Alcoholes Grasos/aislamiento & purificación , Alcoholes Grasos/farmacología , Zingiber officinale/química , Análisis de Varianza , Catecoles/química , Línea Celular , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Cromatografía Líquida de Alta Presión/métodos , Alcoholes Grasos/química , HumanosRESUMEN
Five chicken lines were experimentally infected with a HPAI H7N1 virus, to examine the variation in susceptibility to infection. Three lines showed high susceptibility to the virus, while two showed some resistance, with 7 out of 20, and 11 out of 15 birds, respectively, remaining healthy and surviving the experimental infection. Genotyping for the G/A polymorphism at position 2032 of Mx cDNA showed that one line was fixed for the G allele, and two were segregating for A and G alleles. Birds in the other two lines were selected to be fixed for the A allele. Statistical analyses indicated that the Mx genotype did not affect the clinical status or the time course of infection after viral inoculation.