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1.
Viruses ; 16(6)2024 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-38932224

RESUMO

Porcine parvoviruses (PPVs) are among the most important agents of reproductive failure in swine worldwide. PPVs comprise eight genetically different species ascribed to four genera: Protoparvovirus (PPV1, PPV8), Tetraparvovirus (PPV2-3), Copiparvovirus (PPV4-6), and Chaphamaparvovirus (PPV7). In 2016, PPV7 was firstly detected in the USA and afterwards in Europe, Asia, and South America. Recently, it was also identified in Italy in pig farms with reproductive failure. This study aimed to evaluate the circulation of PPV7 in domestic and wild pigs in Sardinia, Italy. In addition, its coinfection with Porcine Circovirus 2 (PCV2) and 3 (PCV3) was analysed, and PPV7 Italian strains were molecularly characterised. PPV7 was detected in domestic pigs and, for the first time, wild pigs in Italy. The PPV7 viral genome was detected in 20.59% of domestic and wild pig samples. PPV7 detection was significantly lower in domestic pigs, with higher PCV2/PCV3 co-infection rates observed in PPV7-positive than in PPV7-negative domestic pigs. Molecular characterisation of the NS1 gene showed a very high frequency of recombination that could presumably promote virus spreading.


Assuntos
Coinfecção , Infecções por Parvoviridae , Parvovirus Suíno , Filogenia , Doenças dos Suínos , Animais , Parvovirus Suíno/genética , Parvovirus Suíno/classificação , Parvovirus Suíno/isolamento & purificação , Itália/epidemiologia , Infecções por Parvoviridae/veterinária , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/virologia , Suínos , Doenças dos Suínos/virologia , Doenças dos Suínos/epidemiologia , Coinfecção/virologia , Coinfecção/veterinária , Coinfecção/epidemiologia , Genoma Viral , Circovirus/genética , Circovirus/classificação , Circovirus/isolamento & purificação , Infecções por Circoviridae/veterinária , Infecções por Circoviridae/virologia , Infecções por Circoviridae/epidemiologia , DNA Viral/genética
2.
Viruses ; 15(2)2023 01 18.
Artigo em Inglês | MEDLINE | ID: mdl-36851491

RESUMO

Understanding how geography and human mobility shape the patterns and spread of infectious diseases such as COVID-19 is key to control future epidemics. An interesting example is provided by the second wave of the COVID-19 epidemic in Europe, which was facilitated by the intense movement of tourists around the Mediterranean coast in summer 2020. The Italian island of Sardinia is a major tourist destination and is widely believed to be the origin of the second Italian wave. In this study, we characterize the genetic variation among SARS-CoV-2 strains circulating in northern Sardinia during the first and second Italian waves using both Illumina and Oxford Nanopore Technologies Next Generation Sequencing methods. Most viruses were placed into a single clade, implying that despite substantial virus inflow, most outbreaks did not spread widely. The second epidemic wave on the island was actually driven by local transmission of a single B.1.177 subclade. Phylogeographic analyses further suggest that those viral strains circulating on the island were not a relevant source for the second epidemic wave in Italy. This result, however, does not rule out the possibility of intense mixing and transmission of the virus among tourists as a major contributor to the second Italian wave.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , COVID-19/epidemiologia , Epidemiologia Molecular , Itália/epidemiologia , Filogeografia , Variação Genética
3.
Viruses ; 13(11)2021 10 25.
Artigo em Inglês | MEDLINE | ID: mdl-34834952

RESUMO

African swine fever virus (ASFV) is one of the pathogens of highest concern worldwide. Despite different virus lineages co-circulating in several areas, dual infections in the same animal have been rarely observed, suggesting that ASF superinfections are infrequent events. Here we present the first genome-wide detection and analysis of two intragenotype dual ASFV infections. The dual infections have been detected in a hunted wild boar and in a pig carcass, both infected by ASFV genotype I in Sardinia in 1984 and 2018, respectively. We characterize the genetic differences between the two sequences, their intra-host frequency, and their phylogenetic relationship among fully sequenced ASFV strains from Sardinia. Both dual infections involve pairs of closely related but different viruses that were circulating in Sardinia in the same period. The results imply that dual ASFV infections or similar ASFV strains are more common than expected, especially in ASF endemic areas, albeit difficult to detect.


Assuntos
Vírus da Febre Suína Africana/genética , Febre Suína Africana/epidemiologia , Febre Suína Africana/virologia , Surtos de Doenças , Genoma Viral , Genômica , Vírus da Febre Suína Africana/classificação , Animais , Sequência de Bases , DNA Viral/genética , Genótipo , Itália/epidemiologia , Filogenia , Análise de Sequência de DNA , Sus scrofa/virologia , Suínos , Sequenciamento Completo do Genoma
4.
Vaccines (Basel) ; 8(3)2020 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-32967098

RESUMO

African swine fever virus (ASFV), the cause of a devastating disease affecting domestic and wild pigs, has been present in Sardinia since 1978. In the framework of the regional ASF eradication plan, 4484 illegal pigs were culled between December 2017 and February 2020. The highest disease prevalence was observed in the municipality with the highest free-ranging pig density, and culling actions drastically reduced ASFV circulation among these animals. ASFV-antibody were detected in 36.7% of tested animals, which were apparently healthy, thus, the circulation of low-virulence ASFV isolates was hypothesized. ASFV genome was detected in 53 out of 2726 tested animals, and virus isolation was achieved in two distinct culling actions. Two ASFV haemadsorbing strains were isolated from antibody-positive apparently healthy pigs: 55234/18 and 103917/18. Typing analysis revealed that both isolates belong to p72 genotype I, B602L subgroup X; phylogenetic analysis based on whole genome sequencing data showed that they were closely related to Sardinian ASFV strains collected since 2010, especially 22653/Ca/2014. Our data suggested the absence of immune-escaped ASFV variants circulating among free-ranging pigs, indicating that other elements contributed to virus circulation among these animals. Understanding factors behind disease persistence in endemic settings might contribute to developing effective countermeasures against this disease.

5.
Transbound Emerg Dis ; 66(3): 1114-1119, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30715791

RESUMO

African swine fever (ASF) is a notifiable infectious disease, caused by the ASF virus (ASFV), which is a DNA virus belonging to the family Asfarviridae, genus Asfivirus. This disease has gained importance in the last decade after its spread in several countries in Eastern and Central Europe, and more recently, in China. Despite the efforts made to eradicate it, ASF is still present on the Mediterranean island of Sardinia (Italy) and has been since 1978. ASF risk factors on the island have been analysed in previous studies; the role of free-ranging pigs in virus persistence has been suggested, but has not been fully elucidated. The most recent eradication plan provides more stringent measures to combat free-ranging pigs and any kind of illegality in the pig sector. From December 2017 to June 2018, a total of 29 depopulation actions were performed in 13 municipalities in central Sardinia, during which 2,281 free-ranging pigs were culled and more than 50% of them were tested for ASFV and antibody presence (1,218 and 1,416, respectively). A total of 651 pigs were seropositive, with a mean seroprevalence of 53.4% (CI 95% = 50.6-56.3), and 38 were ASFV positive (virus prevalence = 2.6%; CI 95% = 2.1-3.0). To the best of our knowledge, the present study is the first to provide a complete evaluation of this millennial system of pig farming and ASFV prevalence in free-ranging pigs. Furthermore, it has emphasised the necessity of combining the maintenance of an epidemiological surveillance program with continuous education of farmers and other people involved in pig husbandry, based on cultural and economic aspects.


Assuntos
Vírus da Febre Suína Africana/imunologia , Febre Suína Africana/epidemiologia , Erradicação de Doenças , Febre Suína Africana/prevenção & controle , Febre Suína Africana/virologia , Vírus da Febre Suína Africana/genética , Vírus da Febre Suína Africana/isolamento & purificação , Abate de Animais , Animais , Monitoramento Epidemiológico , Fazendas , Feminino , Geografia , Itália/epidemiologia , Masculino , Prevalência , Fatores de Risco , Estudos Soroepidemiológicos , Suínos
6.
J Wildl Dis ; 53(3): 602-606, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28231033

RESUMO

African swine fever (ASF) is one of the most important and complex infectious diseases affecting pigs ( Sus scrofa ). The disease has been present in Sardinia, Italy, since 1978. Factors influencing the presence of the disease on the island are the presence of illegally bred pigs, uncontrolled movements of animals, and local traditions. Implementation of public health programs is essential for controlling ASF. The use of new diagnostic techniques on both wild boar (WB) and illegally bred pigs would provide tools for faster and more inexpensive control of the disease. We evaluated a commercial serological test kit (Pen-side [PS]) for use in the field. We sampled 113 hunter-harvested WB during the 2014-15 season, collecting blood and lung samples to conduct serological analyses and to screen for the ASF virus. Although the sensitivity (81.8%) and specificity (95.9%) of tests performed in the field were reduced compared to the same test in laboratory, they nevertheless allowed for rapid diagnosis and reduced unnecessary carcass destruction. The test, conducted in the field, was less expensive than in the laboratory and required less manpower. Therefore, we conclude that the combined use of antibody PS test and antigen PS test may be a valuable emergency management method during an outbreak as well as a useful tool for conducting regular monitoring activities as a preventive policy.


Assuntos
Vírus da Febre Suína Africana/isolamento & purificação , Febre Suína Africana/diagnóstico , Animais , Surtos de Doenças , Imunoensaio/veterinária , Itália , Sus scrofa/virologia , Suínos
7.
J Enzyme Inhib Med Chem ; 32(1): 264-270, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28097874

RESUMO

3,5-Diaryl-4,5-dihydroisoxazoles were synthesized and evaluated as monoamine oxidase (MAO) enzyme inhibitors and iron chelators. All compounds exhibited selective inhibitory activity towards the B isoform of MAO in the nanomolar concentration range. The best performing compound was preliminarily evaluated for its ability to bind iron II and III cations, indicating that neither iron II nor iron III is coordinated. The best compounds racemic mixtures were separated and single enantiomers inhibitory activity evaluated. Furthermore, none of the synthesised compounds exhibited activity towards MAO A. Overall, these data support our hypothesis that 3,5-diaryl-4,5-dihydroisoxazoles are promising scaffolds for the design of neuroprotective agents.


Assuntos
Isoxazóis/farmacologia , Inibidores da Monoaminoxidase/farmacologia , Monoaminoxidase/metabolismo , Isoxazóis/química , Modelos Moleculares , Inibidores da Monoaminoxidase/química , Relação Estrutura-Atividade
8.
Eur J Med Chem ; 108: 542-552, 2016 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-26717204

RESUMO

With the aim to identify new, potent and selective monoamine oxidase B (MAO-B) inhibitors, molecular interaction field analysis has been applied to a MAO-B complex with 3-acetyl-2,5-diaryl-2,3-dihydro-1,3,4-oxadiazole chemical structure, known as a privileged scaffold for this target. Several compounds displayed potent in vitro activity, exhibiting IC50 values in the medium to low nanomolar range. The enantiomers of most promising derivatives were separated by enantioselective HPLC and in vitro evaluated. Experimental results, according to theoretical drug design, clearly indicated a key role of the ligand stereochemistry in the target recognition/inhibition. In particular the (R)- enantiomers showed the best activity with respect to the (S)- stereoisomer. Finally, docking experiments coupled to molecular dynamics (MD) simulations, were applied for understanding the putative MAO -B binding modes of the new compounds providing detailed information for further structural optimization.


Assuntos
Desenho de Fármacos , Inibidores da Monoaminoxidase/farmacologia , Monoaminoxidase/metabolismo , Oxidiazóis/farmacologia , Simulação por Computador , Relação Dose-Resposta a Droga , Humanos , Simulação de Dinâmica Molecular , Estrutura Molecular , Inibidores da Monoaminoxidase/síntese química , Inibidores da Monoaminoxidase/química , Oxidiazóis/síntese química , Oxidiazóis/química , Relação Estrutura-Atividade
10.
J Med Chem ; 54(18): 6394-8, 2011 Sep 22.
Artigo em Inglês | MEDLINE | ID: mdl-21777011

RESUMO

3-Acetyl-2,5-diaryl-2,3-dihydro-1,3,4-oxadiazoles were designed, synthesized, and tested as inhibitors against human monoamine oxidase (MAO) A and B isoforms. Several compounds, obtained as racemates, were identified as selective MAO-B inhibitors. The enantiomers of some derivatives were separated by enantioselective HPLC and tested. The R-enantiomers always showed the highest activity. Docking study and molecular dynamic simulations demonstrated the putative binding mode. We conclude that these 1,3,4-oxadiazoles derivatives are promising reversible and selective MAO-B inhibitors.


Assuntos
Inibidores da Monoaminoxidase/síntese química , Oxidiazóis/síntese química , Animais , Linhagem Celular , Desenho de Fármacos , Humanos , Insetos/citologia , Isoenzimas/antagonistas & inibidores , Modelos Moleculares , Inibidores da Monoaminoxidase/química , Inibidores da Monoaminoxidase/farmacologia , Oxidiazóis/química , Oxidiazóis/farmacologia , Ligação Proteica , Proteínas Recombinantes/antagonistas & inibidores , Estereoisomerismo , Relação Estrutura-Atividade
11.
Talanta ; 82(1): 426-31, 2010 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-20685489

RESUMO

A direct HPLC enantioseparation of three new chiral oxadiazoline derivatives endowed with potential MAO-B inhibitory activity was accomplished on the immobilised Chiralpak IA chiral stationary phase. Multi-mg amounts of enantiomers with high enantiomeric purity (ee> or =98%) were rapidly collected using pure dichloromethane as eluent. The absolute configuration and chiroptical properties of the enantiomers isolated at semipreparative scale were exhaustively determined.


Assuntos
Amilose/química , Cromatografia Líquida de Alta Pressão/métodos , Inibidores da Monoaminoxidase/isolamento & purificação , Monoaminoxidase/metabolismo , Avaliação Pré-Clínica de Medicamentos , Inibidores da Monoaminoxidase/química , Oxidiazóis/química , Oxidiazóis/isolamento & purificação , Estereoisomerismo
12.
J Pharm Biomed Anal ; 52(5): 665-71, 2010 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-20338705

RESUMO

Analytical and semipreparative high-performance liquid chromatography (HPLC) enantioseparation of the proton-pump inhibitor omeprazole (OME) and its potential organic chiral impurities were accomplished on the immobilised-type Chiralpak IA chiral stationary phase (CSP) under both polar organic and normal-phase conditions. The (S)-enantiomers were isolated with a purity of >99% ee and their absolute configuration was empirically assigned by circular dichroism (CD) spectroscopy. A chemo- and enantioselective HPLC method was validated to control the enantiomeric purity of the (S)-enantiomer of OME (ESO), an active ingredient contained in drug products, in the presence of chiral and achiral related substances. The precision, linearity and accuracy of the determination of the (R)-impurity as well as the recovery of ESO from a pharmaceutical preparation were determined. The proposed method uses the mixture methyl tert-butylether (MtBE)-ethyl acetate (EA)-ethanol (EtOH)-diethylamine (DEA) 60:40:5:0.1 (v/v/v/v) as a mobile phase. In these conditions, linearity over the concentration range 0.5-25 microg/ml for (R)-enantiomer was obtained. The limits of detection and quantification were 99 and 333 ng/ml, respectively. The intra and inter-day assay precision was less than 2% (RSD%).


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Omeprazol/isolamento & purificação , Dicroísmo Circular , Esomeprazol , Limite de Detecção , Omeprazol/química , Padrões de Referência , Reprodutibilidade dos Testes , Espectrofotometria Ultravioleta , Estereoisomerismo
13.
Chirality ; 22(1): 56-62, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19319987

RESUMO

A direct semipreparative HPLC enantioseparation of two chiral thiazolidinone derivatives having cyclooxygenase-2 inhibition activity was performed on the Chiralpak IA chiral stationary phase. Semipreparative amounts of enantiopure forms were collected using acetonitrile-ethanol-trifluoroacetic acid mixtures as mobile phase. The absolute configuration of both compounds was unequivocally established by single-crystal X-ray diffraction method and correlated to the chiroptical properties of isolated enantiomers.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Ciclo-Oxigenase 2/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase/isolamento & purificação , Cristalografia por Raios X , Inibidores de Ciclo-Oxigenase/química , Inibidores de Ciclo-Oxigenase/farmacologia , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Conformação Molecular , Espectrofotometria Ultravioleta , Estereoisomerismo
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