An RT-rtPCR assay for detection of rabies virus in bovine specimens / Um ensaio RT-rtPCR para detecção de vírus da raiva em espécimes bovinos

ABSTRACT: Bovine rabies is endemic in most Brazilian States, including Rio Grande do Sul (RS), which has faced an unprecedented rabies outbreak between 2011 and 2018. We described a real-time reverse transcription quantitative PCR (RT-rtPCR) for detection of rabies virus (RABV) in bovine samples. The primers were designed targeting a highly conserved region of the nucleoprotein (N) gene of RABV obtained from cattle. The detection limit corresponded to 13 DNA copies and the intra- and inter-run repeatability was adequate (CV<9%) in all dilutions tested. Amplification of other pathogens associated with neurological disease in cattle or cross-contamination was not observed. Brain samples from cattle suspicious of rabies (n=21) were tested in triplicate by the RT-rtPCR and by the gold-standard direct fluorescent antibody test (DFAT), resulting in 100% of sensitivity and specificity of the RT-rtPCR. Testing of additional 41 bovine brain samples submitted to the routine DFAT testing yielded 37 (90.2%) concordant results (30 positive/7 negative) and 4 (9.7%) inconclusive in DFAT and RT-rtPCR positive. These results showed a good concordance between the tests and a higher sensitivity of the RT-rtPCR. This assay represents an alternative for RABV detection, either as a confirmatory test or for large-scale diagnosis in endemic regions.

RESUMO: A raiva bovina é endêmica na maioria dos estados brasileiros, inclusive no Rio Grande do Sul (RS), que enfrentou um surto de raiva sem precedentes entre 2011 e 2018. Descrevemos um PCR quantitativo de transcrição reversa em tempo real (RT-rtPCR) para detecção do vírus da raiva (RABV) em bovinos. Os primers foram desenhados visando uma região altamente conservada do gene da nucleoproteína (N) de RABV obtido de bovinos. O limite de detecção correspondeu a 13 cópias de DNA e a repetibilidade intra e inter-ensaios foi adequada (CV <9%) em todas as diluições testadas. Não foi observada amplificação de outros patógenos associados a doenças neurológicas em bovinos ou contaminação cruzada. Amostras de cérebro de bovinos com suspeita de raiva (n = 21) foram testadas em triplicata no RT-rtPCR e pelo teste de anticorpo fluorescente padrão ouro (DFAT), resultando em 100% de sensibilidade e especificidade do RT-rtPCR. O teste de 41 amostras de cérebro bovino adicionais submetidas ao teste de DFAT de rotina rendeu 37 (90,2%) resultados concordantes (30 positivos / sete negativos) e quatro (9,7%) inconclusivos em DFAT e RT-rtPCR positivo. Esses resultados mostraram boa concordância entre os testes e maior sensibilidade do RT-rtPCR. Este ensaio representa uma alternativa para a detecção do vírus da raiva, seja como teste confirmatório ou para diagnóstico em larga escala em regiões endêmicas.

First report of sunflower chlorotic mottle virus infecting sunflower plants in Brazil

Sunflower (Helianthus annuus) plants showing symptoms of chlorosis, mosaic, chlorotic ringspot, and necrosis on younger leaves were found in a small experimental plot in Piracicaba, in the state of São Paulo, Brazil. Preliminary examinations by transmission electron microscopy of symptomatic leaf tissue revealed flexuous filamentous particles 13-15 nm wide and 700-750 nm long, and cytoplasmatic cylindrical inclusions typical of those found in plant cells infected by members of the Potyvirus genus. Total RNA extracted from symptomatic leaves and subjected to RT-PCR followed by partial nucleotide sequencing confirmed the presence of a potyvirus in the affected plants, which was identified as sunflower chlorotic mottle virus (SuCMoV), a member of the Sunflower chlorotic mottle virus (genus Potyvirus, family Potyviridae) species. Mechanical transmission assays with extracts of symptomatic sunflower leaves reproduced the original symptoms in sunflowers, mosaic symptoms in Zinnia elegans, and chlorotic local lesions in Chenopodium amaranticolor and C. quinoa. Sunflower and zinnia plants became infected after aphid transmission experiments with Myzus persicae. RT-PCR tests using specific primers for SuCMoV confirmed the presence of this virus in experimentally infected plants, meeting the criteria of Koch's postulate. This is the first report of SuCMoV infecting sunflower plants in Brazil.

An RT-rtPCR assay for detection of rabies virus in bovine specimens / Um ensaio RT-rtPCR para detecção de vírus da raiva em espécimes bovinos

Bovine rabies is endemic in most Brazilian States, including Rio Grande do Sul (RS), which has faced an unprecedented rabies outbreak between 2011 and 2018. We described a real-time reverse transcription quantitative PCR (RT-rtPCR) for detection of rabies virus (RABV) in bovine samples. The primers were designed targeting a highly conserved region of the nucleoprotein (N) gene of RABV obtained from cattle. The detection limit corresponded to 13 DNA copies and the intra- and inter-run repeatability was adequate (CV<9%) in all dilutions tested. Amplification of other pathogens associated with neurological disease in cattle or cross-contamination was not observed. Brain samples from cattle suspicious of rabies (n=21) were tested in triplicate by the RT-rtPCR and by the gold-standard direct fluorescent antibody test (DFAT), resulting in 100% of sensitivity and specificity of the RT-rtPCR. Testing of additional 41 bovine brain samples submitted to the routine DFAT testing yielded 37 (90.2%) concordant results (30 positive/7 negative) and 4 (9.7%) inconclusive in DFAT and RT-rtPCR positive. These results showed a good concordance between the tests and a higher sensitivity of the RT-rtPCR. This assay represents an alternative for RABV detection, either as a confirmatory test or for large-scale diagnosis in endemic regions.

A raiva bovina é endêmica na maioria dos estados brasileiros, inclusive no Rio Grande do Sul (RS), que enfrentou um surto de raiva sem precedentes entre 2011 e 2018. Descrevemos um PCR quantitativo de transcrição reversa em tempo real (RT-rtPCR) para detecção do vírus da raiva (RABV) em bovinos. Os primers foram desenhados visando uma região altamente conservada do gene da nucleoproteína (N) de RABV obtido de bovinos. O limite de detecção correspondeu a 13 cópias de DNA e a repetibilidade intra e inter-ensaios foi adequada (CV <9%) em todas as diluições testadas. Não foi observada amplificação de outros patógenos associados a doenças neurológicas em bovinos ou contaminação cruzada. Amostras de cérebro de bovinos com suspeita de raiva (n = 21) foram testadas em triplicata no RT-rtPCR e pelo teste de anticorpo fluorescente padrão ouro (DFAT), resultando em 100% de sensibilidade e especificidade do RT-rtPCR. O teste de 41 amostras de cérebro bovino adicionais submetidas ao teste de DFAT de rotina rendeu 37 (90,2%) resultados concordantes (30 positivos / sete negativos) e quatro (9,7%) inconclusivos em DFAT e RT-rtPCR positivo. Esses resultados mostraram boa concordância entre os testes e maior sensibilidade do RT-rtPCR. Este ensaio representa uma alternativa para a detecção do vírus da raiva, seja como teste confirmatório ou para diagnóstico em larga escala em regiões endêmicas.

Eugenol and its liposome-based nano carrier reduce anxiety by inhibiting glyoxylase-1 expression in mice

Abstract The most common form of psycho-social dysfunction is anxiety with depression being related closely without any age bar. They are present with combined state of sadness, confusion, stress, fear etc. Glyoxalase system contains enzyme named glyoxalase 1 (GLO1).It is a metabolic pathway which detoxifies alpha-oxo-aldehydes, particularly methylglyoxal (MG). Methylglyoxal is mainly made by the breakdown of the glycolytic intermediates, glyceraldehyde-3-phosphates and dihydroxyacetone phosphate. Glyoxylase-1 expression is also related with anxiety behavior. A casual role or GLO-1 in anxiety behavior by using viral vectors for over expression in the anterior cingulate cortex was found and it was found that local GLO-1 over expression increased anxiety behavior. The present study deals with the molecular mechanism of protective activity of eugenol against anxiolytic disorder. A pre-clinical animal study was performed on 42 BALB/c mice. Animals were given stress through conventional restrain model. The mRNA expression of GLO-1 was analyzed by real time RT-PCR. Moreover, the GLO-1 protein expression was also examined by immunohistochemistry in whole brain and mean density was calculated. The mRNA and protein expressions were found to be increased in animals given anxiety as compared to the normal control. Whereas, the expressions were decreased in the animals treated with eugenol and its liposome-based nanocarriers in a dose dependent manner. However, the results were better in animals treated with nanocarriers as compared to the compound alone. It is concluded that the eugenol and its liposome-based nanocarriers exert anxiolytic activity by down-regulating GLO-1 protein expression in mice.

Resumo A forma mais comum de disfunção psicossocial é a ansiedade intimamente relacionada com a depressão, sem qualquer barreira de idade. Elas estão presentes em um estado combinado de tristeza, confusão, estresse, medo etc. O sistema de glioxalase contém uma enzima chamada glioxalase 1 (GLO1). É uma via metabólica que desintoxica alfa-oxo-aldeídos, particularmente metilglioxal (MG). O metilglioxal é produzido principalmente pela quebra dos intermediários glicolíticos, gliceraldeído-3-fosfatos e fosfato de diidroxiacetona. A expressão da glioxalase 1 também está relacionada ao comportamento de ansiedade. Um papel casual ou GLO1 no comportamento de ansiedade usando vetores virais para superexpressão no córtex cingulado anterior foi encontrado e descobriu-se que a superexpressão local de GLO1 aumentava o comportamento de ansiedade. O presente estudo trata do mecanismo molecular da atividade protetora do eugenol contra o transtorno ansiolítico. Um estudo pré-clínico em animais foi realizado em 42 camundongos BALB / c. Os animais foram submetidos ao estresse por meio do modelo de contenção convencional. A expressão de mRNA de GLO1 foi analisada por RT-PCR em tempo real. Além disso, a expressão da proteína GLO1 também foi examinada por imuno-histoquímica em todo o cérebro e a densidade média foi calculada. Verificou-se que as expressões de mRNA e proteínas estavam aumentadas em animais que receberam ansiedade em comparação com o controle normal. Considerando que as expressões foram diminuídas nos animais tratados com eugenol e seus nanocarreadores baseados em lipossomas de forma dependente da dose. No entanto, os resultados foram melhores em animais tratados com nanocarreadores em comparação com o composto sozinho. Conclui-se que o eugenol e seus nanocarreadores baseados em lipossomas exercem atividade ansiolítica por regulação negativa da expressão da proteína GLO1 em camundongos.

Incidence of Begomovirus and Crinivirus in tomato and potato crops in Paraná state, Brazil / Incidência de Begomovirus e Crinivirus nas culturas do tomateiro e batateira no estado do Paraná, Brasil

In the last decades, the high incidence of viruses transmitted by whiteflies has become a problem in the tomato fields, threatening, more recently, the potato crops. The present study carried out a survey of begomoviruses and criniviruses in tomato and potato crops, from 2015 to 2018, in the municipalities of Araucária, Campo do Tenente, Campo Largo, Contenda, Lapa, Faxinal, Morretes, Reserva, Castro, Palmeira and São Mateus do Sul, in Paraná State, Brazil. Total DNA and RNA from leaves were extracted and used as templates to detect, respectively, begomoviruses by PCR and criniviruses by RT-PCR. Out of 215 tomato samples, 14 from Faxinal were infected by crinivirus. The other tomato samples and 243 potato samples showed negative results for begomovirus and crinivirus. Results indicated a low incidence (6.5%) of crinivirus infecting tomato crops in Paraná State, and the nucleotide sequence of one amplified fragment shared 99.71% identity with tomato chlorotic virus (ToCV).

Nas últimas décadas, a alta incidência de vírus transmitidos por mosca-branca tornou-se um problema nos campos de tomateiros, ameaçando, mais recentemente, a cultura da batateira. O presente trabalho teve como objetivo realizar um levantamento de begomovírus e crinivírus em lavouras de tomateiro e batateira nos municípios de Araucária, Campo do Tenente, Campo Largo, Contenda, Lapa, Faxinal, Morretes, Reserva, Castro, Palmeira e São Mateus do Sul, no Estado do Paraná, Brasil, de 2015 à 2018. DNA e RNA totais de folhas foram extraídos e utilizados como molde para detectar begomovírus por PCR e crinivírus por RT-PCR. Das 215 amostras de tomateiros coletadas, 14 provenientes de Faxinal estavam infectadas por crinivírus. As demais amostras de tomateiro e as 243 amostras de batateira analisadas apresentaram resultados negativo para begomovírus e crinivírus. Os resultados indicaram baixa incidência (6,5%) de crinivírus infectando lavouras de tomateiros no Estado do Paraná e a sequência de nucleotídeos de um amplicon apresentou 99,71% de identidade com o crinivírus tomato chlorotic virus (ToCV).

Molecular detection and prevalence of Rotavirus with acute gastroenteritis among the children of rural and urban areas / Detecção molecular e prevalência de rotavírus com gastroenterite aguda entre as crianças de áreas rurais e urbanas

Rotavirus is the main infective agent of acute gastroenteritis (AGE) in children under the age of five years and causing significant morbidity as well as mortality throughout the world. The study was carried out to detect the prevalence rate, genotypes strain and risk factors of Rotavirus among the children of rural and urban areas of district Bannu Khyber Pakhtunkhwa Pakistan. A total of 180 stool samples were collected from children under the age of 5 years from two major hospitals of Bannu from January to December (2015). The samples were analyzed by Reverse-transcriptase Polymerase Chain Reaction (RT-PCR) for the detection of Rotavirus, positive samples were further processed for genotyping (G and P type) through specific PCR. Of the total, 41 (23%) samples were positive for Rotavirus. The most prevalent G genotypes found were: G3, G8, G9 (each 29%), followed by G10 (15%), and G11 (10%). Whereas the prevalent P genotypes were: P-8 (25%), P-4 and P-10 (each 20%), P-9 (15%), followed by P-6 and P-11 (each 10%). Moreover, Rotavirus infection was more prevalent in summer (23.73%) and winter (22.7%) than spring (20%) and autumn (21.4%). Rotavirus infection exhibited high frequency in June (14%), October (8%) and November (6%). It is concluded that Rotavirus is more prevalent in children and various genotypes (G and P) of Rotavirus are present in the study area. Lack of studies, awareness and rarer testing of Rotavirus are the principal reasons of virus prevalence in district Bannu, Pakistan.

O rotavírus é o principal agente infeccioso da gastroenterite aguda (AGE) em crianças menores de 5 anos e causa de morbidade e mortalidade significativas em todo o mundo. O estudo foi realizado para detectar a taxa de prevalência, cepa de genótipos e fatores de risco de rotavírus entre as crianças de áreas rurais e urbanas do distrito de Bannu Khyber Pakhtunkhwa, Paquistão. Um total de 180 amostras de fezes foi coletada de crianças menores de 5 anos de dois grandes hospitais de Bannu de janeiro a dezembro (2015). As amostras foram analisadas por reação em cadeia da polimerase transcriptase reversa (RT-PCR) para detecção de rotavírus; as amostras positivas foram posteriormente processadas para genotipagem (tipo G e P) através de PCR específica. Do total, 41 (23%) amostras foram positivas para rotavírus. Os genótipos G mais prevalentes encontrados foram: G3, G8, G9 (cada 29%), seguidos de G10 (15%) e G11 (10%). Considerando que os genótipos P prevalentes foram: P-8 (25%), P-4 e P-10 (cada 20%), P-9 (15%), seguido por P-6 e P-11 (cada 10%). Além disso, a infecção por rotavírus foi mais prevalente no verão (23,73%) e inverno (22,7%) do que na primavera (20%) e no outono (21,4%). A infecção por rotavírus apresentou alta frequência em junho (14%), outubro (8%) e novembro (6%). Conclui-se que o rotavírus é mais prevalente em crianças e vários genótipos (G e P) do rotavírus estão presentes na área de estudo. A falta de estudos, conhecimento e testes mais raros de rotavírus são as principais razões da prevalência do vírus no distrito de Bannu, Paquistão.

Molecular detection and prevalence of Rotavirus with acute gastroenteritis among the children of rural and urban areas / Detecção molecular e prevalência de rotavírus com gastroenterite aguda entre as crianças de áreas rurais e urbanas

Rotavirus is the main infective agent of acute gastroenteritis (AGE) in children under the age of five years and causing significant morbidity as well as mortality throughout the world. The study was carried out to detect the prevalence rate, genotypes strain and risk factors of Rotavirus among the children of rural and urban areas of district Bannu Khyber Pakhtunkhwa Pakistan. A total of 180 stool samples were collected from children under the age of 5 years from two major hospitals of Bannu from January to December (2015). The samples were analyzed by Reverse-transcriptase Polymerase Chain Reaction (RT-PCR) for the detection of Rotavirus, positive samples were further processed for genotyping (G and P type) through specific PCR. Of the total, 41 (23%) samples were positive for Rotavirus. The most prevalent G genotypes found were: G3, G8, G9 (each 29%), followed by G10 (15%), and G11 (10%). Whereas the prevalent P genotypes were: P-8 (25%), P-4 and P-10 (each 20%), P-9 (15%), followed by P-6 and P-11 (each 10%). Moreover, Rotavirus infection was more prevalent in summer (23.73%) and winter (22.7%) than spring (20%) and autumn (21.4%). Rotavirus infection exhibited high frequency in June (14%), October (8%) and November (6%). It is concluded that Rotavirus is more prevalent in children and various genotypes (G and P) of Rotavirus are present in the study area. Lack of studies, awareness and rarer testing of Rotavirus are the principal reasons of virus prevalence in district Bannu, Pakistan.(AU)

O rotavírus é o principal agente infeccioso da gastroenterite aguda (AGE) em crianças menores de 5 anos e causa de morbidade e mortalidade significativas em todo o mundo. O estudo foi realizado para detectar a taxa de prevalência, cepa de genótipos e fatores de risco de rotavírus entre as crianças de áreas rurais e urbanas do distrito de Bannu Khyber Pakhtunkhwa, Paquistão. Um total de 180 amostras de fezes foi coletada de crianças menores de 5 anos de dois grandes hospitais de Bannu de janeiro a dezembro (2015). As amostras foram analisadas por reação em cadeia da polimerase transcriptase reversa (RT-PCR) para detecção de rotavírus; as amostras positivas foram posteriormente processadas para genotipagem (tipo G e P) através de PCR específica. Do total, 41 (23%) amostras foram positivas para rotavírus. Os genótipos G mais prevalentes encontrados foram: G3, G8, G9 (cada 29%), seguidos de G10 (15%) e G11 (10%). Considerando que os genótipos P prevalentes foram: P-8 (25%), P-4 e P-10 (cada 20%), P-9 (15%), seguido por P-6 e P-11 (cada 10%). Além disso, a infecção por rotavírus foi mais prevalente no verão (23,73%) e inverno (22,7%) do que na primavera (20%) e no outono (21,4%). A infecção por rotavírus apresentou alta frequência em junho (14%), outubro (8%) e novembro (6%). Conclui-se que o rotavírus é mais prevalente em crianças e vários genótipos (G e P) do rotavírus estão presentes na área de estudo. A falta de estudos, conhecimento e testes mais raros de rotavírus são as principais razões da prevalência do vírus no distrito de Bannu, Paquistão.(AU)

Epizootiology of canine distemper in naturally infected dogs in Goiânia, Brazil / Epizootiologia da cinomose em cães naturalmente infectados em Goiânia, Brasil

Although the epizootiological profile of canine distemper in Goiânia is unknown, there is clinical evidence for a high incidence of canine distemper virus (CDV) infection among dogs. Therefore, this study determined the epizootiological characteristics of canine distemper in naturally infected dogs. Data of 46 dogs that tested positive for the CDV based on immunochromatography or reverse transcription-polymerase chain reaction were collected. Data on the sex, breed, age, and vaccination status were obtained from these dogs, and extraneural and neural sign analyses were performed. Although, the infected dogs belonged to both sexes, different breeds, and different age groups, a greater proportion of cases were seen in adults (1-6 years), undefined breeds, and unvaccinated dogs. Among the CDV-positive dogs, 10.87% had been vaccinated. In addition, 4.35% showed neural signs, 8.69% showed extraneural signs, and 86.96% showed both. High lethality was observed, with viral antigen and/or DNA detected in 82.61% dead dogs. Only 8.70% of the total CDV-infected dogs remained alive at the time of their assessment.

Embora o perfil epizootiológico da cinomose canina em Goiânia seja desconhecido, há evidencia clínica para alta incidência da infecção pelo vírus da cinomose (CDV) nos cães. Este estudo objetivou determinar as características epizootiológicas da cinomose em cães naturalmente infectados. Dados de 46 cães positivos por imunocromatografia ou reação em cadeia da polimerase via transcriptase reversa para o CDV foram coletados. Dados sobre sexo, raça, idade, estado vacinal foram obtidos desses cães, e os sinais extraneurais e neurais foram analisados. Animais de ambos os sexos, diferentes raças e idades foram acometidos. A maior proporção de casos foi vista em adultos (de um a seis anos), sem raça definida e não vacinados. Dentre os cães positivos, 10,87% haviam sido vacinados. Em adição, 4,35% apresentaram sinais neurais, 8,69% sinais extraneurais e 86,96% mostraram ambos. Alta letalidade foi observada, com o antígeno viral e/ou DNA identificado em 82,61% dos cães que foram a óbito. Apenas 8,7% dos cães infectados permaneceram vivos até o momento da avaliação.

Molecular detection and prevalence of Rotavirus with acute gastroenteritis among the children of rural and urban areas

Abstract Rotavirus is the main infective agent of acute gastroenteritis (AGE) in children under the age of five years and causing significant morbidity as well as mortality throughout the world. The study was carried out to detect the prevalence rate, genotypes strain and risk factors of Rotavirus among the children of rural and urban areas of district Bannu Khyber Pakhtunkhwa Pakistan. A total of 180 stool samples were collected from children under the age of 5 years from two major hospitals of Bannu from January to December (2015). The samples were analyzed by Reverse-transcriptase Polymerase Chain Reaction (RT-PCR) for the detection of Rotavirus, positive samples were further processed for genotyping (G and P type) through specific PCR. Of the total, 41 (23%) samples were positive for Rotavirus. The most prevalent G genotypes found were: G3, G8, G9 (each 29%), followed by G10 (15%), and G11 (10%). Whereas the prevalent P genotypes were: P-8 (25%), P-4 and P-10 (each 20%), P-9 (15%), followed by P-6 and P-11 (each 10%). Moreover, Rotavirus infection was more prevalent in summer (23.73%) and winter (22.7%) than spring (20%) and autumn (21.4%). Rotavirus infection exhibited high frequency in June (14%), October (8%) and November (6%). It is concluded that Rotavirus is more prevalent in children and various genotypes (G and P) of Rotavirus are present in the study area. Lack of studies, awareness and rarer testing of Rotavirus are the principal reasons of virus prevalence in district Bannu, Pakistan.

Resumo O rotavírus é o principal agente infeccioso da gastroenterite aguda (AGE) em crianças menores de 5 anos e causa de morbidade e mortalidade significativas em todo o mundo. O estudo foi realizado para detectar a taxa de prevalência, cepa de genótipos e fatores de risco de rotavírus entre as crianças de áreas rurais e urbanas do distrito de Bannu Khyber Pakhtunkhwa, Paquistão. Um total de 180 amostras de fezes foi coletada de crianças menores de 5 anos de dois grandes hospitais de Bannu de janeiro a dezembro (2015). As amostras foram analisadas por reação em cadeia da polimerase transcriptase reversa (RT-PCR) para detecção de rotavírus; as amostras positivas foram posteriormente processadas para genotipagem (tipo G e P) através de PCR específica. Do total, 41 (23%) amostras foram positivas para rotavírus. Os genótipos G mais prevalentes encontrados foram: G3, G8, G9 (cada 29%), seguidos de G10 (15%) e G11 (10%). Considerando que os genótipos P prevalentes foram: P-8 (25%), P-4 e P-10 (cada 20%), P-9 (15%), seguido por P-6 e P-11 (cada 10%). Além disso, a infecção por rotavírus foi mais prevalente no verão (23,73%) e inverno (22,7%) do que na primavera (20%) e no outono (21,4%). A infecção por rotavírus apresentou alta frequência em junho (14%), outubro (8%) e novembro (6%). Conclui-se que o rotavírus é mais prevalente em crianças e vários genótipos (G e P) do rotavírus estão presentes na área de estudo. A falta de estudos, conhecimento e testes mais raros de rotavírus são as principais razões da prevalência do vírus no distrito de Bannu, Paquistão.

Eugenol and its liposome-based nano carrier reduce anxiety by inhibiting glyoxylase-1 expression in mice / Eugenol e seu nanocarreador à base de lipossoma reduzem a ansiedade ao inibir a expressão de glioxalase 1 em camundongos

The most common form of psycho-social dysfunction is anxiety with depression being related closely without any age bar. They are present with combined state of sadness, confusion, stress, fear etc. Glyoxalase system contains enzyme named glyoxalase 1 (GLO1).It is a metabolic pathway which detoxifies alpha-oxo-aldehydes, particularly methylglyoxal (MG). Methylglyoxal is mainly made by the breakdown of the glycolytic intermediates, glyceraldehyde-3-phosphates and dihydroxyacetone phosphate. Glyoxylase-1 expression is also related with anxiety behavior. A casual role or GLO-1 in anxiety behavior by using viral vectors for over expression in the anterior cingulate cortex was found and it was found that local GLO-1 over expression increased anxiety behavior. The present study deals with the molecular mechanism of protective activity of eugenol against anxiolytic disorder. A pre-clinical animal study was performed on 42 BALB/c mice. Animals were given stress through conventional restrain model. The mRNA expression of GLO-1 was analyzed by real time RT-PCR. Moreover, the GLO-1 protein expression was also examined by immunohistochemistry in whole brain and mean density was calculated. The mRNA and protein expressions were found to be increased in animals given anxiety as compared to the normal control. Whereas, the expressions were decreased in the animals treated with eugenol and its liposome-based nanocarriers in a dose dependent manner. However, the results were better in animals treated with nanocarriers as compared to the compound alone. It is concluded that the eugenol and its liposome-based nanocarriers exert anxiolytic activity by down-regulating GLO-1 protein expression in mice.

A forma mais comum de disfunção psicossocial é a ansiedade intimamente relacionada com a depressão, sem qualquer barreira de idade. Elas estão presentes em um estado combinado de tristeza, confusão, estresse, medo etc. O sistema de glioxalase contém uma enzima chamada glioxalase 1 (GLO1). É uma via metabólica que desintoxica alfa-oxo-aldeídos, particularmente metilglioxal (MG). O metilglioxal é produzido principalmente pela quebra dos intermediários glicolíticos, gliceraldeído-3-fosfatos e fosfato de diidroxiacetona. A expressão da glioxalase 1 também está relacionada ao comportamento de ansiedade. Um papel casual ou GLO1 no comportamento de ansiedade usando vetores virais para superexpressão no córtex cingulado anterior foi encontrado e descobriu-se que a superexpressão local de GLO1 aumentava o comportamento de ansiedade. O presente estudo trata do mecanismo molecular da atividade protetora do eugenol contra o transtorno ansiolítico. Um estudo pré-clínico em animais foi realizado em 42 camundongos BALB / c. Os animais foram submetidos ao estresse por meio do modelo de contenção convencional. A expressão de mRNA de GLO1 foi analisada por RT-PCR em tempo real. Além disso, a expressão da proteína GLO1 também foi examinada por imuno-histoquímica em todo o cérebro e a densidade média foi calculada. Verificou-se que as expressões de mRNA e proteínas estavam aumentadas em animais que receberam ansiedade em comparação com o controle normal. Considerando que as expressões foram diminuídas nos animais tratados com eugenol e seus nanocarreadores baseados em lipossomas de forma dependente da dose. No entanto, os resultados foram melhores em animais tratados com nanocarreadores em comparação com o composto sozinho. Conclui-se que o eugenol e seus nanocarreadores baseados em lipossomas exercem atividade ansiolítica por regulação negativa da expressão da proteína GLO1 em camundongos.

Eugenol and its liposome-based nano carrier reduce anxiety by inhibiting glyoxylase-1 expression in mice / Eugenol e seu nanocarreador à base de lipossoma reduzem a ansiedade ao inibir a expressão de glioxalase 1 em camundongos

The most common form of psycho-social dysfunction is anxiety with depression being related closely without any age bar. They are present with combined state of sadness, confusion, stress, fear etc. Glyoxalase system contains enzyme named glyoxalase 1 (GLO1).It is a metabolic pathway which detoxifies alpha-oxo-aldehydes, particularly methylglyoxal (MG). Methylglyoxal is mainly made by the breakdown of the glycolytic intermediates, glyceraldehyde-3-phosphates and dihydroxyacetone phosphate. Glyoxylase-1 expression is also related with anxiety behavior. A casual role or GLO-1 in anxiety behavior by using viral vectors for over expression in the anterior cingulate cortex was found and it was found that local GLO-1 over expression increased anxiety behavior. The present study deals with the molecular mechanism of protective activity of eugenol against anxiolytic disorder. A pre-clinical animal study was performed on 42 BALB/c mice. Animals were given stress through conventional restrain model. The mRNA expression of GLO-1 was analyzed by real time RT-PCR. Moreover, the GLO-1 protein expression was also examined by immunohistochemistry in whole brain and mean density was calculated. The mRNA and protein expressions were found to be increased in animals given anxiety as compared to the normal control. Whereas, the expressions were decreased in the animals treated with eugenol and its liposome-based nanocarriers in a dose dependent manner. However, the results were better in animals treated with nanocarriers as compared to the compound alone. It is concluded that the eugenol and its liposome-based nanocarriers exert anxiolytic activity by down-regulating GLO-1 protein expression in mice.(AU)

A forma mais comum de disfunção psicossocial é a ansiedade intimamente relacionada com a depressão, sem qualquer barreira de idade. Elas estão presentes em um estado combinado de tristeza, confusão, estresse, medo etc. O sistema de glioxalase contém uma enzima chamada glioxalase 1 (GLO1). É uma via metabólica que desintoxica alfa-oxo-aldeídos, particularmente metilglioxal (MG). O metilglioxal é produzido principalmente pela quebra dos intermediários glicolíticos, gliceraldeído-3-fosfatos e fosfato de diidroxiacetona. A expressão da glioxalase 1 também está relacionada ao comportamento de ansiedade. Um papel casual ou GLO1 no comportamento de ansiedade usando vetores virais para superexpressão no córtex cingulado anterior foi encontrado e descobriu-se que a superexpressão local de GLO1 aumentava o comportamento de ansiedade. O presente estudo trata do mecanismo molecular da atividade protetora do eugenol contra o transtorno ansiolítico. Um estudo pré-clínico em animais foi realizado em 42 camundongos BALB / c. Os animais foram submetidos ao estresse por meio do modelo de contenção convencional. A expressão de mRNA de GLO1 foi analisada por RT-PCR em tempo real. Além disso, a expressão da proteína GLO1 também foi examinada por imuno-histoquímica em todo o cérebro e a densidade média foi calculada. Verificou-se que as expressões de mRNA e proteínas estavam aumentadas em animais que receberam ansiedade em comparação com o controle normal. Considerando que as expressões foram diminuídas nos animais tratados com eugenol e seus nanocarreadores baseados em lipossomas de forma dependente da dose. No entanto, os resultados foram melhores em animais tratados com nanocarreadores em comparação com o composto sozinho. Conclui-se que o eugenol e seus nanocarreadores baseados em lipossomas exercem atividade ansiolítica por regulação negativa da expressão da proteína GLO1 em camundongos.(AU)

Hepatitis C virus genotype diversity in Ningxia Hui autonomous region, Northwestern China / Diversidade genética do vírus da hepatite C na região autônoma de Ningxia Hui

Hepatitis C virus (HCV) genotypes vary greatly in different regions. The aim of this study is to investigate the distribution of HCV genotypes in HCV infected patients, in Ningxia Hui Autonomous Region. Nucleic acid extraction and amplification were performed with test kits on 153 HCV infected patients serum samples. The HCV viral load was measured using reverse transcriptase PCR (RT-PCR) and HCV genotypes were determined. Among the 153 HCV-infected patients, 56 had genotype (GT)1b (36.60%), 45 had GT2a (29.40%), 23 had GT3a (15.00%), 14 had GT3b (9.20%),13 had GT6a (8.50%), 1 had GT1g (0.70%), 1 had GT6xa (0.70%). In GT1b, 21.40% were female and 78.60% were male; in GT2a, 42.20% were female and 57.80% were male;Males were most prevalent in genotypes 1b(39.30%), while female were most prevalent in genotype 2a(46.30%). Rare GT1g and GT6xa were also detected in males. The 41-50 year age group had the highest HCV prevalence of 32.00%. HCV GT1b is the predominant HCV genotype in Ningxia Hui Autonomous Region.

Os genótipos do vírus da hepatite C (HCV) variam muito de acordo com a região. O objetivo deste estudo é investigar a distribuição do genótipo do vírus da hepatite C em pessoas infectadas pelo vírus na região autônoma de Ningxia Hui. A extração de ácido nucleico e a expansão de amostras séricas em 153 pacientes infectados com HCV foram realizadas utilizando kits de ensaio. A capacidade do vírus HCV foi medida pela reação em cadeia da polimerase retrotranscrição (RT-PCR) e o genótipo HCV foi determinado. Os genótipos (Gt) tiveram a seguinte distribuição entre os 153 casos de infecção HCV: 56 Gt 1-B (36, 60%), 45 Gt 2-A (29, 40%), 23 Gt 3-A (15, 00%), 14 Gt 3-B (9, 20%), 13 Gt 6-A (8, 50%), 1 Gt 1-G (0, 70%) e 1 Gt 6-Xa (0, 70%). Já o sexo dos indivíduos teve a seguinte distribuição: Gt 1-B, 21, 40% mulheres e 78, 60% homens; Gt 2-A, 42, 20% mulheres e 57, 80% homens. A presença de homens é mais comum no genótipo 1-B (39, 30%), enquanto as mulheres ocorrem mais comumente no genótipo 2-A (46, 30%). Gt 1-G e Gt 6-Xa raros também foram detectados em homens. A taxa de infecção com HCV para grupos etários de 41 a 50 anos é a mais alta, com 32,00%. HCV Gt 1-B é o genótipo dominante do HCV na região autônoma de Ningxia Hui.

Eugenol and its liposome-based nano carrier reduce anxiety by inhibiting glyoxylase-1 expression in mice / Eugenol e seu nanocarreador à base de lipossoma reduzem a ansiedade ao inibir a expressão de glioxalase 1 em camundongos

Abstract The most common form of psycho-social dysfunction is anxiety with depression being related closely without any age bar. They are present with combined state of sadness, confusion, stress, fear etc. Glyoxalase system contains enzyme named glyoxalase 1 (GLO1).It is a metabolic pathway which detoxifies alpha-oxo-aldehydes, particularly methylglyoxal (MG). Methylglyoxal is mainly made by the breakdown of the glycolytic intermediates, glyceraldehyde-3-phosphates and dihydroxyacetone phosphate. Glyoxylase-1 expression is also related with anxiety behavior. A casual role or GLO-1 in anxiety behavior by using viral vectors for over expression in the anterior cingulate cortex was found and it was found that local GLO-1 over expression increased anxiety behavior. The present study deals with the molecular mechanism of protective activity of eugenol against anxiolytic disorder. A pre-clinical animal study was performed on 42 BALB/c mice. Animals were given stress through conventional restrain model. The mRNA expression of GLO-1 was analyzed by real time RT-PCR. Moreover, the GLO-1 protein expression was also examined by immunohistochemistry in whole brain and mean density was calculated. The mRNA and protein expressions were found to be increased in animals given anxiety as compared to the normal control. Whereas, the expressions were decreased in the animals treated with eugenol and its liposome-based nanocarriers in a dose dependent manner. However, the results were better in animals treated with nanocarriers as compared to the compound alone. It is concluded that the eugenol and its liposome-based nanocarriers exert anxiolytic activity by down-regulating GLO-1 protein expression in mice.

Resumo A forma mais comum de disfunção psicossocial é a ansiedade intimamente relacionada com a depressão, sem qualquer barreira de idade. Elas estão presentes em um estado combinado de tristeza, confusão, estresse, medo etc. O sistema de glioxalase contém uma enzima chamada glioxalase 1 (GLO1). É uma via metabólica que desintoxica alfa-oxo-aldeídos, particularmente metilglioxal (MG). O metilglioxal é produzido principalmente pela quebra dos intermediários glicolíticos, gliceraldeído-3-fosfatos e fosfato de diidroxiacetona. A expressão da glioxalase 1 também está relacionada ao comportamento de ansiedade. Um papel casual ou GLO1 no comportamento de ansiedade usando vetores virais para superexpressão no córtex cingulado anterior foi encontrado e descobriu-se que a superexpressão local de GLO1 aumentava o comportamento de ansiedade. O presente estudo trata do mecanismo molecular da atividade protetora do eugenol contra o transtorno ansiolítico. Um estudo pré-clínico em animais foi realizado em 42 camundongos BALB / c. Os animais foram submetidos ao estresse por meio do modelo de contenção convencional. A expressão de mRNA de GLO1 foi analisada por RT-PCR em tempo real. Além disso, a expressão da proteína GLO1 também foi examinada por imuno-histoquímica em todo o cérebro e a densidade média foi calculada. Verificou-se que as expressões de mRNA e proteínas estavam aumentadas em animais que receberam ansiedade em comparação com o controle normal. Considerando que as expressões foram diminuídas nos animais tratados com eugenol e seus nanocarreadores baseados em lipossomas de forma dependente da dose. No entanto, os resultados foram melhores em animais tratados com nanocarreadores em comparação com o composto sozinho. Conclui-se que o eugenol e seus nanocarreadores baseados em lipossomas exercem atividade ansiolítica por regulação negativa da expressão da proteína GLO1 em camundongos.

Distinct carbapenems susceptibility profiles in isogenic isolates of Klebsiella pneumoniae presenting Ompk36 disruption and expression of down-regulated blaKPC-2 / Perfis distintos de suscetibilidade a carbapenêmicos em isolados isogênicos de Klebsiella pneumoniae apresentando interrupção de Ompk36 e expressão de blaKPC-2 negativamente regulada

The isolation of multidrug-resistant Klebsiella pneumoniae in hospitals is a major public health threat, increasing patient hospitalization costs, morbidity and mortality. Therefore, this work investigated the resistance mechanisms that produced different carbapenems susceptibility profiles in two isogenic strains of K. pneumoniae isolated from the same patient in a public hospital in Recife, Pernambuco. The genes that encode the main porins in K. pneumoniae, ompK35 and ompK36, and several beta-lactamase genes were analyzed. The expression of these genes was evaluated by quantitative real time PCR (polymerase chain reaction) with reverse transcriptase (RT-qPCR). SDS-PAGE (sodium dodecyl sulphate­polyacrylamide gel electrophoresis) was performed to analyze the outer membrane proteins. The analysis of the ompK36 genetic environment disclosed an IS903 insertion sequence disrupting this gene in the ertapenem resistant isolate (KPN133). The blaKPC-2 gene showed down-regulated expression in both isolates. Our findings show that changes in porins, especially OmpK36, are more determinant to carbapenems susceptibility profile of bacterial isolates than variations in blaKPC gene expression.

O isolamento de Klebsiella pneumoniae multirresistente em hospitais é uma grande ameaça à saúde pública, aumentando os custos de internação, morbidade e mortalidade dos pacientes. Portanto, este trabalho investigou os mecanismos de resistência que produziram diferentes perfis de suscetibilidade aos carbapenêmicos em duas cepas isogênicas de K. pneumoniae isoladas do mesmo paciente em um hospital público em Recife, Pernambuco. Foram analisados ​​os genes que codificam as principais porinas em K. pneumoniae, ompK35 e ompK36, e diversos genes de beta-lactamases. A expressão desses genes foi avaliada por PCR (reação em cadeia da polimerase quantitativa em tempo real) com transcriptase reversa (RT-qPCR). SDS-PAGE (dodecil sulfato de sódio-poliacrilamida gel eletroforese) foi realizada para analisar as proteínas da membrana externa. A análise do ambiente genético ompK36 revelou uma sequência de inserção IS903 interrompendo este gene no isolado resistente ao ertapenem (KPN133). O gene blaKPC-2 apresentou expressão negativamente regulada em ambos os isolados. Nossos achados mostram que alterações nas porinas, especialmente OmpK36, são mais determinantes no perfil de suscetibilidade aos carbapenêmicos de isolados bacterianos do que variações na expressão do gene blaKPC.

Extensive emphysematous pyelonephritis in a nondiabetic female cat - treatment with unilateral nephroureterectomy

Background: Emphysematous pyelonephritis (EPN) is an acute, severe necrotizing infection of the renal parenchyma and surrounding tissues that results in gas formation in the kidney, collecting system, or surroundings. EPN is a rare condition in veterinary medicine and occurs most frequently in dogs with diabetes mellitus. Although the prognosis of medical management in animals is poor, the standardized treatment protocol according to EPN severity is unclear. This report describes the first case of a nondiabetic female cat with extensive EPN and good prognosis following direct nephroureterectomy (NU). Case: A 10-year-old spayed female cat presented with the chief complaint of an acute loss of weight within 1 week, vomiting, and disorientation including stumbling, discoordination, circling, wobbling, head tilting, and difficulties in standing. At presentation, the patient had a body condition score of 1/9 and weighed 2.6 kg. Blood examination revealed leukocytosis, anemia, and hypoproteinemia. Abdominal radiography revealed severely decreased serosal details. A massive gas silhouette observed in the peritoneal and retroperitoneal cavities, was diagnosed as abdominal free gas. Abdominal ultrasound showed an accumulation of moderately anechoic fluid mixed with gas and cyst-like capsules around the left kidney. Left partial ureteral obstruction and dilation were also observed. Computed tomography (CT) was performed without sedatives or anesthetic drugs. The findings showed severe inflammatory changes in the peritoneum and a loss of the normal inner structure in the left kidney. A pyelogram of the left kidney was not observed after injection of the contrast material. Diffuse fat stranding and free gas observed in the mesentery of the entire abdominal cavity and around the left kidney were considered septic peritonitis. Urinalysis revealed proteinuria and hematuria. Numerous neutrophils with rod-type bacteria were observed in the ascites. Following diagnostic examinations, the patient was diagnosed with extensive left EPN, including inflammatory ascites and abdominal free gas. Therefore, emergency NU of the nonfunctional left kidney and ruptured ureter and thorough abdominal lavage were conducted. Diffuse inflammation and a nephrolith were observed in the section of the harvested kidney. The nephrolith was composed of 100% calcium oxalate monohydrate. The realtime polymerase chain reaction (RT-PCR) test for feline infectious peritonitis (FIP) was negative. Escherichia coli was detected in the ascites, and antibiotic therapy was administered following the antibiotic sensitivity test. The histological findings from the left kidney and ureter included marked chronic inflammation and fibrosis. The patient was discharged 4 days after surgery. During the 8-month follow-up period, the patient's condition improved. Discussion: This was a unique case of EPN in a nondiabetic cat and the first reported case of EPN with a ruptured ureter, including abdominal free gas, inflammatory ascites, and peritonitis. This patient had a bacterial urinary tract infection with E. coli, which is the most frequently isolated pathogen in humans. This gas-forming bacteria produced a massive amount of gas and inflammation that were considered to have ruptured the urinary tract, so that the gas was released into the abdomen. This case corresponded to class 3B, with two risk factors according to the human EPN classification system. Direct NU and abdominal lavage were performed as emergency surgeries. The patient stabilized gradually and showed a good prognosis. Immediate surgical intervention is recommended in animal patients showing the extensive EPN stage.

Early maturation of corpus luteum in rabbits - effect of sildenafil citrate on luteolytic capacity in the early luteal period

Background: Prostaglandin F2 alpha (PGF2 α) binds to the specific receptor (PTGFR) on the corpus luteum (CL) in mammals, inducing regression of the CL structure (luteolysis) and initiating a new cycle. While PGF2 α is effective only on mature CL, the immature CL structure (early luteal phase) resists PGF2 α. In this study, sildenafil citrate, which is used to increase blood flow in the genital organs for treating specific pregnancy issues in women, was administered during the early luteal phase in a rabbit model to test the hypothesis of enhancing blood flow to the CL, thereby promoting earlier maturation and enabling a response to PGF2 α. Materials, Methods & Results: The study was conducted in 2 sub-studies: clinical and molecular. A large number of rabbits were initially included in the sub-studies to ensure a sufficient number of pseudo-pregnant rabbits. Ovulation in rabbits was induced with buserelin acetate and was considered as day 0 of the study. The sub-studies were continued with rabbits whose pseudo-pregnancies were confirmed according to progesterone (P4 ) results. As a result, the studies were continued with a total of 41 pseudo-pregnant New Zealand female rabbits, 21 of which were included in the clinical sub-study and 20 in the molecular sub-study. In both sub-studies, on day 3 of the luteal period, rabbits in the treatment group received 5 mg/kg sildenafil citrate and all rabbits received a single dose of exogenous PGF2 α on day 4 to induce luteolysis. In the clinical sub-study, echotexture and intraovarian blood flow changes in the ovaries were determined by ultrasonography (USG) examination. In the molecular sub-study, the expression changes of Hypoxia Inducible Factor 1 Alpha (HIF1A) and Vascular Endothelial Growth Factor (VEGF) related to angiogenesis, Steroidogenic Acute Regulatory Protein (StAR) related to P4 metabolism, Prostaglandin-Endoperoxide Synthase 2 (PTGS2) related to prostaglandin (PG) mechanism and 15-Hydroxyprostaglandin Dehydrogenase (HPGD) genes at mRNA level were determined using Real Time Polymerase Chain Reaction (RT-PCR) in CL tissues obtained with ovariohysterectomy (OVH) at 1 and 12 h after PGF2 α injection. In addition, blood samples were collected for determine P4 levels from all rabbits. In the clinical sub-study; there was no difference between the groups in mean gray values (MGV), whereas there was a significant decrease in both pulsatile index (PI) and resistance index (RI) values at 40 min after PGF2 α injection (P < 0.05). In the molecular sub-study, it was determined that sildenafil citrate had no significant effect (P > 0.05) on the expression levels 1 and 12 h after PGF2 α injection. According to the results of the molecular sub-study, no significant effect of sildenafil citrate on the mRNA expression levels in the investigated genes was detected (P > 0.05). However, within each group, differences were found according to OVH time after PGF2 α injection. It was observed that PTGS2 and HPGD mRNA expressions decreased at the 12th h compared to the 1st h, while HIF1A expression increased (P < 0.05). Discussion: According to the results obtained from clinical and molecular sub-studies, it was determined that a single dose of sildenafil citrate (5 mg/kg) applied on the 3rd day of the luteal period did not contribute to the maturation process of the CL, did not increase blood flow, and was insufficient to break the resistance of the CL against PGF2 α applied on the 4th day of the luteal period. However, a significant decrease in the PI value at the 40th min after PGF2 α injection suggests that sildenafil citrate has a supportive effect, and that this decrease is also seen in the RI value, suggesting that its effect is insufficient against the vasoconstrictive effect of PGF2 α.

Bovine viral diarrhea virus infection in cattle - antioxidant status and some biochemical parameters

Background: Bovine viral diarrhea virus (BVDV) infections in cattle result in significant economic losses due to reproductive performance deficiencies caused by gastrointestinal, respiratory system infections, and transplacental infections. BVDV is one of the most important and widespread pathogens in cattle worldwide, including Turkey. Methods such as virus neutralization, enzyme-linked immunosorbent assay (ELISA), reverse transcriptase and polymerase chain reaction (RT-PCR) are used for the detection of the disease. The diagnosis of the disease in its subclinical form is challenging due to the lengthy and costly procedures involved. Investigating oxidative stress parameters in ruminants with various diseases contributes significantly to diagnosis and prognosis. This study aimed to investigate some oxidative stress and biochemical parameters in cattle infected with BVDV. Materials, Methods & Results: In the study, blood samples were collected from 80 Simmental breed cows aged between approximately 4 and 8 years to determine the presence of BVDV antibodies using the ELISA method. Based on the results obtained, study groups were organized. The study included a group of 10 animals with positive antibody levels as the infected group, and a group of 10 animals with negative antibody levels as the healthy group. Blood samples were taken from the animals, and serum separation was ensured. In the obtained serum samples, levels of vitamin E, vitamin A, ß-Carotene, catalase, GSH-Px, and MDA were determined using spectrophotometric methods. In addition, serum total protein, albumin, alkaline phosphatase (ALP), aspartate aminotransferase (AST), glucose, low-density lipoprotein (LDL), high-density lipoprotein (HDL), calcium (Ca), and phosphorus (P) were measured using commercial test kits and an autoanalyzer. In the study, it was observed that the differences in serum MDA, vitamin E, vitamin A, ß-carotene, and catalase levels were statistically significant between the healthy and BVDV-infected groups (P < 0.001). The activity of GSH-Px was also found to be statistically different between the groups (P < 0.01). Among the biochemical parameters, HDL, LDL, and AST levels were found to be statistically significant between the healthy and BVDV-infected groups (P < 0.001). Additionally, ALP and glucose levels were found to be statistically significant (P < 0.01). However, although there were differences in the levels of total protein, albumin, Ca, and P between the groups, these results were not statistically significant. Discussion: Although the diagnosis of the disease was partially made based on clinical observations in BVDV infections, the ELISA method was used for accurate diagnosis. Furthermore, it was found that there was a significant difference in MDA concentration between the healthy and infected groups, indicating oxidative damage caused by the virus. Similarly, significant differences in vitamin E, vitamin A, ß-carotene, GSH-Px, and catalase levels were observed between the groups, indicating a decrease in antioxidant values due to the infection. In addition, differences in ALP, AST, glucose, LDL, and HDL levels were found between the groups. This difference is thought to be related to the effects of the disease agent on the liver and systemically. This study demonstrates that, in addition to the viral pathogen, antioxidant and biochemical values are important criteria in the detection of the disease.

Gender-based incidence, recovery period, and mortality rate of COVID-19 among the population of district Attock, Pakistan / Incidência baseada em gênero, período de recuperação e taxa de mortalidade de COVID-19 entre a população do distrito de Attock, Paquistão

Abstract COVID-19 is reported as an extremely contagious disease with common symptoms of fever, dry cough, sore throat, and tiredness. The published literature on incidence and gender-wise prevalence of COVID-19 is scarce in Pakistan. Therefore, the present study was designed to compare the distribution, incubation period and mortality rate of COVID-19 among the male and female population of district Attock. The data were collected between 01 April 2020 and 07 December 2020 from the population of district Attock, Pakistan. A total of 22,962 individuals were screened and 843 were found positive for RT-qPCR for SARS-CoV-2. The confirmed positive cases were monitored carefully. Among the positive cases, the incidence of COVID-19 was 61.7% among males and 38.2% among females. The average recovery period of males was 18.89±7.75 days and females were 19±8.40 days from SARS-CoV-2. The overall mortality rate was 8.06%. The death rate of male patients was significantly higher (P<0.05) compared to female patients. Also, the mortality rate was higher (P<0.05) in male patients of 40-60 years of age compared to female patients of the same age group. Moreover, the mortality rate significantly increased (P<0.05) with the increase of age irrespective of gender. In conclusion, the incidence and mortality rate of COVID-19 is higher in males compared to the female population. Moreover, irrespective of gender the mortality rate was significantly lower among patients aged <40 years.

Resumo Covid-19 é relatada como uma doença extremamente contagiosa com sintomas comuns de febre, tosse seca, dor de garganta e cansaço. A literatura publicada sobre incidência e prevalência de Covid-19 com base no gênero é escassa no Paquistão. Portanto, o presente estudo teve como objetivo comparar a distribuição, o período de incubação e a taxa de mortalidade de Covid-19 entre a população masculina e feminina do distrito de Attock. Os dados foram coletados entre 1 de abril de 2020 e 7 de dezembro de 2020 da população do distrito de Attock, Paquistão. Um total de 22.962 indivíduos foi selecionado, e 843 foram considerados positivos para RT-qPCR para SARS-CoV-2. Os casos positivos confirmados foram monitorados cuidadosamente. Entre os casos positivos, a incidência de Covid-19 foi de 61,7% no sexo masculino e 38,2% no feminino. O período médio de recuperação dos homens foi de 18,89 ± 7,75 dias e das mulheres 19 ± 8,40 dias do SARS-CoV-2. A mortalidade geral foi de 8,06%. A taxa de mortalidade de pacientes do sexo masculino foi significativamente maior (P < 0,05) em comparação com pacientes do sexo feminino. Além disso, a taxa de mortalidade foi maior (P < 0,05) em pacientes do sexo masculino com 40-60 anos de idade em comparação com pacientes do sexo feminino da mesma faixa etária. Além disso, a taxa de mortalidade aumentou significativamente (P < 0,05) com o aumento da idade, independentemente do sexo. Em conclusão, a incidência e a taxa de mortalidade de Covid-19 são maiores no sexo masculino em comparação com a população feminina. E também, independentemente do sexo, a taxa de mortalidade foi significativamente menor entre os pacientes com idade < 40 anos.

Persistent SARS-CoV-2 antigen presence in multiple organs of a naturally infected cat from Brazil

Abstract Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiological agent of the disease coronavirus 2019 (COVID-19) in humans. SARS-CoV-2 has been identified in cats with or without clinical signs. Case presentation: We describe the pathological and molecular findings in a six-month-old asymptomatic cat with SARS-CoV-2 infection from Brazil, belonging to a human family with COVID-19 cases. The pool of nasopharynx and oropharynx swabs at day zero tested positive by RT-qPCR for SARS-CoV-2. No amplification resulted from molecular testing performed on days 7 and 14. The cat was hit by a car and died 43 days after the molecular diagnosis. Immunohistochemistry at post-mortem examination demonstrated nucleocapsid protein in samples from the lungs, kidneys, nasal conchae, trachea, intestine, brain and spleen. Conclusion: The present study has highlighted the possibility that viral antigens can be detected by immunohistochemistry in multiple organs six weeks after infection, although the same tissues tested negative by RT-PCR.

Detection of respiratory viruses and expression of inflammatory cytokines in patients with acute exacerbation chronic obstructive pulmonary disease in Mongolia China

Abstract Chronic obstructive pulmonary disease (COPD) was estimated to be the third cause of global mortality by 2020. Acute exacerbation COPD (AECOPD) is a sudden worsening of COPD symptoms and could be due to virus/bacterial infections and air pollution. Increased expression of inflammatory markers in patients with AECOPD is associated with viral infection. This study aimed to detect different viruses and analyze the expression of various inflammatory markers associated with AECOPD patients. Three hundred and forty-seven patients diagnosed with COPD according to GOLD criteria were included in this study. Swab samples and blood were collected for the detection of viruses by RT-PCR and expression of inflammatory markers, respectively. Of the swab samples, 113 (32.6%) of samples were positive for virus detection. Of these, HRV (39.8%) was the predominant virus detected followed by FluB (27.4%) and FluA (22.1%). The presence of HRV was significantly higher (p=0.044) among the other detected viruses. When compared to healthy controls the expression levels of TNF-, IL-6 and IL-8 were significantly higher (p 0.05) in virus-positive patients. The IL-6 and IL-8 were the next predominantly expressed in markers among the samples. The higher expression rate of IL-8 was significantly (p 0.05) associated with patients having COPD GOLD III severity level and smoking history. Although HRV was the predominant virus detected the combined prevalence of Influenza A and B surpassing the rate of HRV. The high-level expression of well known inflammatory markers of AECOPD, TNF-, IL-6 and IL-8 indicates a chronic severe illness. These markers play an important role and could be used as a marker for determining the severity of AECOPD.

Resumo Estima-se que a doença pulmonar obstrutiva crônica (DPOC) seja a terceira causa de mortalidade global em 2020. A exacerbação aguda DPOC (AECOPD) é um agravamento súbito dos sintomas da DPOC e pode ser devido a infecções por vírus/bactérias e poluição do ar. O aumento da expressão de marcadores inflamatórios em pacientes com AECOPD está associado à infecção viral. Este estudo teve como objetivo detectar diferentes vírus e analisar a expressão de vários marcadores inflamatórios associados a pacientes com AECOPD. Trezentos e quarenta e sete pacientes com diagnóstico de DPOC de acordo com os critérios GOLD foram incluídos neste estudo. Amostras de swab e sangue foram coletadas para detecção de vírus por RT-PCR e expressão de marcadores inflamatórios, respectivamente. Das amostras de esfregaço, 113 (32,6%) amostras foram positivas para detecção de vírus. Nestas, o HRV (39,8%) foi o vírus predominante detectado, seguido do FluB (27,4%) e do FluA (22,1%). A presença de VFC foi significativamente maior (p = 0,044) entre os demais vírus detectados. Quando comparados a controles saudáveis, os níveis de expressão de TNF-, IL-6 e IL-8 foram significativamente maiores (p 0,05) em pacientes com vírus positivo. A IL-6 e a IL-8 foram as próximas predominantemente expressas em marcadores entre as amostras. A maior taxa de expressão de IL-8 foi significativamente (p 0,05) associada a pacientes com grau de gravidade GOLD III da DPOC e história de tabagismo. Embora o HRV tenha sido o vírus predominante, a prevalência combinada de Influenza A e B ultrapassou a taxa de HRV. O alto nível de expressão de marcadores inflamatórios bem conhecidos de AECOPD, TNF-, IL-6 e IL-8 indica uma doença crônica grave. Esses marcadores desempenham um papel importante e podem ser usados como um marcador para determinar a gravidade da AECOPD.