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Use of smartphone messaging applications to increase diagnostic efficiency in veterinary diagnostic laboratories
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Lemos, Ricardo A.A.

; Guizelini, Carolina C.

; Silva, Thaísa X.

; Souza, Larissa L.

; Fonseca, Helena C.F.

; Bonato, Gisele C.

; Gomes, Danilo C.

; Pupin, Rayane C.

Pesquisa Veterinária Brasileira

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2023, Volume 43 elocation e07226

Abstract: En Pt | Text: En Pt | PDF: En | PDF: Pt

ABSTRACT: Many inconclusive diagnoses have been reported in studies carried out in veterinary diagnostic laboratories (VDLs). These inconclusive diagnoses are often associated with field veterinarians sending tissue samples and carcasses for histopathological examination. The use of social media in care has the potential as a tool for improvements in animal health. We evaluated the improvement in the rate of conclusive diagnoses in the VDL when using a messaging application for smartphones as a guidance tool for field veterinarians to refer animals and sample tissues for necropsy and histopathology. Veterinarians contacted the VDL team via a messaging app and forwarded videos, images, and clinical-epidemiological history. Based on this information, guidelines were created in real-time for contacting professionals about methods for collecting samples during necropsy and making diagnoses and scheduling visits to properties. The data obtained showed that smartphone messaging applications could be useful tools to expand the performance of VDLs and improve their diagnostic efficiency, especially when sending samples of animals that died is impossible due to certain sanitary conditions.

RESUMO: Muitos diagnósticos inconclusivos têm sido relatados em estudos realizados em laboratórios de diagnóstico veterinário (LDVs). Esses diagnósticos inconclusivos são frequentemente associados a veterinários de campo que enviam amostras de tecidos e carcaças para exame histopatológico. O uso das mídias sociais com cuidado tem potencial como ferramenta para melhorias na saúde animal. Avaliamos a melhora na taxa de diagnósticos conclusivos no LDV ao utilizar um aplicativo de mensagens para smartphones como ferramenta de orientação para veterinários de campo encaminharem animais e amostras de tecidos para necropsia e histopatologia. Os veterinários entraram em contato com a equipe do L por meio de um aplicativo de mensagens e encaminharam vídeos, imagens e histórico clínico-epidemiológico. Com base nessas informações, foram criadas orientações em tempo real para contato dos profissionais sobre métodos de coleta de amostras durante a necropsia e realização de diagnósticos, além do agendamento de visitas às propriedades. Os dados obtidos mostraram que os aplicativos de mensagens para smartphones podem ser ferramentas úteis para ampliar o desempenho dos VDLs e melhorar sua eficiência diagnóstica, principalmente quando o envio de amostras de animais que morreram é impossível devido a determinadas condições sanitárias.

https://doi.org/10.1590/1678-5150-pvb-7226

Differentials in the epidemiological profile of canine visceral leishmaniasis in the semi-arid region of Paraíba, Brazil
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Silva, Raizza B.S.

; Franco-Silva, Laysa F.

; Lima, Diana A.

; Fonseca, Saul S.

; Ferreira, Marília A.S.

; Silva, Rafael C.

; Lira, Milena A.A.

; Melo, Marcia A.

Pesquisa Veterinária Brasileira

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2021, Volume 41 elocation e06773

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

RESUMO: O objetivo desta pesquisa foi estimar a prevalência da leishmaniose visceral canina (LVC) e identificar as diferenças nos fatores relacionados à sua ocorrência nas zonas urbana e rural do município de Santa Luzia, localizado no semiárido paraibano. Nos anos de 2015 e 2016, coletaram-se 779 amostras de sangue de cães. A prevalência foi determinada através de três técnicas sorológicas, Kit ELISA-S7®, teste rápido DPP® e Kit EIE-LVC®, considerando positivas as amostras que reagiram em, pelo menos, dois ensaios. Os fatores relacionados foram determinados por meio das análises estatísticas uni e multivariada das respostas dos tutores ao questionário epidemiológico. A prevalência de anticorpos anti-Leishmania infantum encontrada no município estudado foi de 15,00% (117/779), sendo maior na zona urbana (15,20%) do que na rural (13,60%). O bairro que apresentou maior prevalência foi o Frei Damião com 26,40% (33/125), sendo considerado um hotspot (OR 1,245; p=0,007). Outros fatores relacionados encontrados foram a criação semidomiciliar (OR 1,798; p=0,025), na zorna urbana, e cão de caça (OR 18,505; p=0,016), contato com bovinos (OR 17,298; p=0,022) e ambiente onde o cão é criado (OR 4,802; p=0,024), na zona rural. Verifica-se a elevada prevalência da leishmaniose visceral canina e a sua ampla distribuição no município de Santa Luzia. Diferenças epidemiológicas entre as zonas urbana e rural puderam ser observadas, demonstrando a necessidade de medidas de controle mais adequadas para cada localidade e comprovando o processo de urbanização.

ABSTRACT: The objective of this study was to estimate the prevalence of canine visceral leishmaniasis (CVL) and to identify the differences in associated factors to its occurrence in urban area and countrysides in the municipality of Santa Luzia located in the semi-arid region of Paraíba. In the years 2015 and 2016, 779 blood samples from dogs were collected. The prevalence was determined by three serological techniques, ELISA-S7® Kit, DPP® Rapid Test and EIE-LVC® Kit, considering positive the samples that reacted in at least two assays. Associated factors were determined by univariate and multivariate analyzes of the guardians’ responses to the epidemiological questionnaire. The prevalence of anti-Leishmania infantum antibodies in the studied municipality was 15.00% (117/779), being higher in the urban area (15.20%) than in the countryside (13.60%). The neighborhood with the highest prevalence was Frei Damião with 26.40% (33/125), being considered a hotspot (OR 1.245, p=0.007). Other associated factors were the semi-domiciliary breeding (OR 1.798, p=0.025), in the urban area, and hunting dog (OR 18.505, p=0.016), contact with cattle (OR 17.298, p=0.022) and environment where the dog is raised (OR 4.802, p=0.024) in the countryside. In the municipality of Santa Luzia, the prevalence of canine visceral leishmaniasis is high and the disease is widely distributed. Epidemiological differences between urban area and the countryside could be observed demonstrating the need for more adequate control measures for each locality and proving the urbanization process.

https://doi.org/10.1590/1678-5150-pvb-6773 25 downloads

α,β-Amyrin prevents steatosis and insulin resistance in a high-fat diet-induced mouse model of NAFLD via the AMPK-mTORC1-SREBP1 signaling mechanism
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Lima, R.P. de

; Nunes, P.I.G.

; Viana, A.F.S.C.

; Oliveira, F.T.B. de

; Silva, R.A.C.

; Alves, A.P.N.N.

; Viana, D.A.

; Fonseca, S.G.C.

; Carvalho, A.A.

; Chaves, M.H.

; Rao, V.S.

; Santos, F.A.

Brazilian Journal of Medical and Biological Research

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2021, Volume 54 N. 10 elocation e11391

Abstract: En | Text: En | PDF: En

Nonalcoholic fatty liver disease (NAFLD), characterized by hepatosteatosis and steatohepatitis, is intrinsically related to obesity. Our previous study reported on the anti-obese activity of α,β-amyrin (AMY), a pentacyclic triterpene isolated from Protium heptaphyllum. This study investigated its ability to prevent fatty liver and the underlying mechanism using the mouse model of NAFLD. NAFLD was induced in male Swiss mice fed a high fat diet (HFD) for 15 weeks. The controls were fed a normal chow diet (ND). The mice were simultaneously treated with AMY at 10 and 20 mg/kg or fenofibrate at 50 mg/kg. Lipid levels along with metabolic and inflammatory parameters were assessed in liver and serum. The liver sections were histologically examined using H&E staining. RT-qPCR and western blotting assays were performed to analyze signaling mechanisms. Mice fed HFD developed severe hepatic steatosis with elevated triglycerides and lipid droplets compared with ND controls. This was associated with a decrease in AMP-activated protein kinase (AMPK) activity, an increase of mechanistic target of rapamycin complex 1 (mTORC1) signaling, and enhanced sterol regulatory element binding protein 1 (SREBP1) expression, which have roles in lipogenesis, inhibition of lipolysis, and inflammatory response. AMY treatment reversed these signaling activities and decreased the severity of hepatic steatosis and inflammatory response, evidenced by serum and liver parameters as well as histological findings. AMY-induced reduction in hepatic steatosis seemed to involve AMPK-mTORC1-SREBP1 signaling pathways, which supported its beneficial role in the prevention and treatment of NAFLD.

https://doi.org/10.1590/1414-431x2021e11391 194 downloads

Anti-inflammatory and antinociceptive effects of the isatin derivative (Z)-2-(5-chloro-2-oxoindolin-3-ylidene)-N-phenyl-hydrazinecarbothioamide in mice
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Dantas, L.L.S.F.R.

; Fonseca, A.G.

; Pereira, J.R.

; Furtado, A.A.

; Gomes, P.A.T.M.

; Fernandes-Pedrosa, M.F.

; Leite, A.C.L.

; Rêgo, M.J.B.M.

; Pitta, M.G.R.

; Lemos, T.M.A.M.

Brazilian Journal of Medical and Biological Research

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2020, Volume 53 N. 10 elocation e10204

Abstract: En | Text: En | PDF: En

Several isatin derivatives have shown important biological activities, which have attracted interest from researchers. For this reason, the present study aimed to evaluate the anti-inflammatory and antinociceptive effects of the isatin derivative (Z)-2-(5-chloro-2-oxoindolin-3-ylidene)-N-phenyl-hydrazinecarbothioamide (COPHCT) in mice. Three doses of this compound were tested: 1.0, 2.5, and 5.0 mg/kg. The anti-inflammatory activity was assessed using the carrageenan-induced paw edema model and the zymosan-induced air pouch model. The evaluation of the antinociceptive effect was performed through the formalin test and the acetic acid-induced abdominal writhing test. The paw edema assay demonstrated that all doses of the compound showed a significant reduction of the edema in the second hour evaluated, but a better response was observed in the fourth hour. The zymosan-induced air pouch model indicated that the compound, in all doses, significantly reduced leukocyte migration and total protein concentration levels. In the formalin test, the doses 1.0, 2.5, and 5.0 mg/kg of COPHCT showed activity only in the second phase, with reduction in paw pain time of 73.61, 79.46, and 73.85%, respectively. The number of abdominal writhings decreased with the increasing dose, but only 5.0 mg/kg COPHCT exhibited a significant response, with a reduction of 24.88%. These results demonstrated the ability of this compound to interfere in the anti-inflammatory activity of edema, vascular permeability, and cell migration. In addition, its possible antinociceptive effect may be related to the dose used.

https://doi.org/10.1590/1414-431x202010204 882 downloads

Molecular detection of bovine immunodeficiency virus (BIV) in bovines from the state of Minas Gerais, Brazil
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Rodrigues, A.P.S.

; Fonseca Júnior, A.A.

; Lima, G.K.

; Bicalho, J.M.

; Leite, R.C.

; Reis, J.K.P.

Arquivo Brasileiro de Medicina Veterinária e Zootecnia

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apr 2019, Volume 71 N. 2 Pages 711 - 714

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

RESUMO O vírus da imunodeficiência bovina é o agente causador da imunodeficiência viral bovina que é conhecido por infectar bovinos em todo o mundo. Como em outras infecções por retrovírus, os hospedeiros desenvolvem uma infecção de longo prazo e a maioria dos animais infectados permanece assintomática. O objetivo deste estudo foi detectar DNA proviral BIV em amostras de sangue de bovinos e estimar a ocorrência de infecção no estado de Minas Gerais, Brasil. Amostras de sangue de 391 bovinos foram coletadas de duas regiões do estado, Zona da Mata e Central. O DNA proviral foi detectado por reação em cadeia da polimerase semi-nested (SN-PCR). Os resultados de SN-PCR indicaram uma ocorrência de BIV de 12,5% no estado. Os produtos amplificados foram confirmados como BIV por sequenciamento e a similaridade da sequência de nucleotídeos com a estirpe de referência (R-29) foi de 99%. Este é o primeiro estudo que relata a presença do BIV em Minas Gerais, Brasil. Os resultados indicam a necessidade de realizar um estudo detalhado sobre a prevalência da infecção por BIV no Brasil.

https://doi.org/10.1590/1678-4162-10495 861 downloads

Late emergence of A594V and L595W mutations related to ganciclovir resistance in a patient with HCMV retinitis and long-term HIV progression
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Slavov, S.N.

; Vilar, F.C.

; Wagatsuma, V.M.D.

; Santana, R.C.

; Machado, A.A.

; Fonseca, B.A.L. da

; Kashima, S.

; Covas, D.T.

Brazilian Journal of Medical and Biological Research

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sep 2015, Volume 48 N. 9 Pages 777 - 781

Abstract: En | Text: En | PDF: En

The emergence of ganciclovir (GCV) resistance during the treatment of human cytomegalovirus (HCMV) infection is a serious clinical challenge, and is associated with high morbidity and mortality. In this case report, we describe the emergence of two consecutive mutations (A594V and L595W) related to GCV resistance in a patient with HCMV retinitis and long-term HIV progression after approximately 240 days of GCV use. Following the diagnosis of retinitis, the introduction of GCV did not result in viral load reduction. The detected mutations appeared late in the treatment, and we propose that other factors (high initial HCMV load, previous GCV exposure, low CD4+ cell count), in addition to the presence of resistance mutations, may have contributed to the treatment failure of HCMV infection in this patient.

https://doi.org/10.1590/1414-431x20154507 1250 downloads

Diagnóstico etiológico de enfermidades do sistema nervoso central de equinos no Estado de Minas Gerais, Brasil
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Costa, E.A.

; Rosa, R.

; Oliveira, T.S.

; Furtini, R.

; Fonseca Júnior, A.A.

; Paixão, T.A.

; Santos, R.L.

Arquivo Brasileiro de Medicina Veterinária e Zootecnia

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apr 2015, Volume 67 N. 2 Pages 391 - 399

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

O Brasil possui o quarto maior rebanho equino do mundo, e o Estado de Minas Gerais detém o maior número de equinos do país. Portanto, um diagnóstico preciso das doenças neurológicas dos equinos é prioridade no estado. Sendo assim, o objetivo deste estudo foi identificar, utilizando a Reação em Cadeia pela Polimerase (PCR), os agentes infecciosos responsáveis por enfermidades que afetam o sistema nervoso central (SNC) de equinos. De janeiro de 2009 a janeiro de 2011, foi realizado um levantamento dos casos de encefalites e encefalomielites em equinos no Estado de Minas Gerais, utilizando-se amostras de SNC de equinos que morreram com sinais neurológicos. Das 217 amostras de SNC, 47 (21,7%) foram positivas para o vírus da raiva pelo método de imunofluorescência indireta e inoculação em camundongos. Nas 170 amostras negativas para o vírus da raiva, o herpes-vírus equino-1 (EHV-1) foi diagnosticado em 20 (11,8%) e o herpes-vírus suíno-1 (SHV-1), em uma amostra por meio de PCR, e o vírus encefalite de Saint Louis (SLEV), em outra amostra, através de transcrição reversa (RT) e PCR (RT-PCR). Constatou-se que o vírus da raiva é o principal agente causador de encefalite em equinos, apesar do crescente número de casos de encefalomielite associados ao EHV-1 no Estado de Minas Gerais.

Brazil has the fourth largest equine herd in the world and the State of Minas Gerais has the largest equine population in the country. Therefore, an accurate diagnosis of cases of neurologic diseases is a priority in Minas Gerais. The aim of this study was to identify by Polymerase Chain Reaction (PCR) infectious agents associated with neurological disease in the central nervous system (CNS) of horses. A survey of encephalitis and encephalomyelitis in horses in Minas Gerais State was performed on samples of CNS from horses that died with neurological signs from January 2009 to January 2011. Forty seven CNS samples from 217 (21.7%) horses were positive for rabies virus by the indirect immunofluorescence assay and mouse inoculation. Among the 170 samples that were negative for rabies, EHV-1 was detected in 20 (11.8%) and the swine herpesvirus-1 (SHV-1) was detected in one sample by PCR, and the Saint Louis encephalitis virus (SLEV) was identified in another sample by reverse transcription (RT) and PCR (RT-PCR). Rabies virus is the most common causative agent of encephalitis in horses, despite the increasing number of cases of encephalitis associated with EHV-1 in the State of Minas Gerais.

https://doi.org/10.1590/1678-6765 6199 downloads

Viral suppression and adherence among HIV-infected children and adolescents on antiretroviral therapy: results of a multicenter study
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Cruz, Maria L.S.

; Cardoso, Claudete A.A.

; Darmont, Mariana Q.

; Souza, Edvaldo

; Andrade, Solange D.

; D'Al Fabbro, Marcia M.

; Fonseca, Rosana

; Bellido, Jaime G.

; Monteiro, Simone S.

; Bastos, Francisco I.

Jornal de Pediatria

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dec 2014, Volume 90 N. 6 Pages 563 - 571

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

OBJETIVO: Avaliar a adesão ao tratamento antirretroviral entre portadores de HIV acompanhados em centros pediátricos. MÉTODOS: Trata-se de estudo transversal multicêntrico. Os prontuários ambulatoriais foram revistos e aplicadas escala de adesão, avaliação de qualidade de vida (WHOQOL-BREF), ansiedade, depressão e uso indevido de álcool/substâncias entre cuidadores. Os desfechos incluíram autorrelato 100% de adesão nos últimos três dias e carga viral do HIV (CV) < 50 cópias/mL. RESULTADOS: 260 indivíduos foram incluídos, 79% crianças e 21% adolescentes; 93% das crianças e 77% dos adolescentes relataram 100% de adesão; 57% das crianças e 49% dos adolescentes tinham CV < 50 cópias /mL. Na análise univariada, diagnóstico do HIV por triagem devido à infecção materna, cuidador com pontuação menor para ansiedade e maior nos domínios físico e psicológico do WHOQOL-BREF se mostraram independentemente associados a 100% de adesão. Intervalos mais curtos entre visitas de farmácia foram associados com CV < 50 cópias /mL (p ≤ 0,01). Regressão multivariada mostrou que os cuidadores sem abuso de álcool/outras drogas (OR = 0,49; IC95% 0,27-0,89) e o intervalo médio entre visitas de farmácia < 33 dias (OR = 0,97; IC95% 0,95-0,98) foram associados com CV < 50 cópias/mL; cuidador com menores escores para ansiedade (OR = 2,57; IC95% 1,27-5,19) e diagnóstico de crianças por triagem devido à infecção materna (OR = 2,25; IC95% 1,12-4,50) foram associados com 100% de adesão. CONCLUSÕES: Programas de HIV pediátrico devem avaliar qualidade de vida e sintomas de ansiedade e depressão dos cuidadores. Registros de farmácia são essenciais na identificação de adesão insatisfatória.

OBJECTIVE: To evaluate treatment adherence among perinatally-infected pediatric human immunodeficiency virus (HIV) patients followed in pediatric centers in Brazil. METHODS: This was a cross-sectional multicenter study. Medical records were reviewed and adherence scale, assessment of caregivers' quality of life (WHOQOL-BREF), anxiety, depression, and alcohol/substances use/abuse were assessed. Outcomes included self-reported 100% adherence in the last three days and HIV viral load (VL) < 50 copies/mL. Statistical analyses included contingency tables and respective statistics, and multivariable logistic regression. RESULTS: 260 subjects were enrolled: 78% children and 22% adolescents; 93% of caregivers for the children and 77% of adolescents reported 100% adherence; 57% of children and 49% of adolescents had VL < 50 copies/mL. In the univariate analyses, HIV diagnosis for screening due to maternal infection, lower caregiver scores for anxiety, and higher scores in physical and psychological domains of WHOQOL-BREF were associated with 100% adherence. Shorter intervals between pharmacy visits were associated with VL < 50 copies/mL (p ≤ 0.01). Multivariable regression demonstrated that caregivers who did not abuse alcohol/other drugs (OR = 0.49; 95% CI: 0.27-0.89) and median interval between pharmacy visits < 33 days (OR = 0.97; 95% CI: 0.95-0.98) were independently associated with VL < 50 copies/mL; whereas lower caregiver scores for anxiety (OR = 2.57; 95% CI: 1.27-5.19) and children's HIV diagnosis for screening due to maternal infection (OR = 2.25; 95% CI: 1.12-4.50) were found to be independently associated with 100% adherence. CONCLUSIONS: Pediatric HIV programs should perform routine assessment of caregivers' quality of life, and anxiety and depression symptoms. In this setting, pharmacy records are essential to help identify less-than-optimal adherence.

https://doi.org/10.1016/j.jped.2014.04.007 5998 downloads

PCR em tempo real para detecção do vírus da doença de Aujeszky
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Fonseca Júnior, A.A.

; Cotorello, A.C.

; Dias, N.L.

; D'Ambros, R.

; Leite, R.C.

; Heneimann, M.B.

; Reis, J.K.P.

Arquivo Brasileiro de Medicina Veterinária e Zootecnia

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jun 2013, Volume 65 N. 3 Pages 801 - 808

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

O objetivo deste trabalho foi desenvolver uma PCR em tempo real (qPCR) para o diagnóstico rápido e sensível da doença de Aujeszky. Os iniciadores amplificaram um fragmento de 123 pares de base do gene codificante da glicoproteína D. A qPCR foi testada em 25 amostras de cérebro de suíno positivas e 85 amostras negativas para DA no isolamento viral e na soroneutralização. A sensibilidade analítica foi calculada com acréscimo de um isolado brasileiro do SuHV-1 titulado em amostras de cérebro de suíno negativas na soroneutralização e na PCR. A técnica apresentou sensibilidade analítica de 10-0,5 TCID50/50µL. A qPCR foi capaz de distinguir reações inespecíficas devido a dímero de oligonucleotídeos iniciadores ou amplificações, além do alvo designado (evitando, assim, os falso-positivos), e de obter resultados rápidos.

The aim of this study was to validate a low-cost real-time PCR for a quick and sensitive diagnosis of the disease. The fluorofore used was a DNA intercalating agent, one of the cheaper detection systems. Primers amplified a 123 base pairs fragment of the gene coding for glycoprotein D. PCR was tested on 25 samples of pig brain positive for AD and 85 samples negative in viral isolation and serum neutralization. The detection limit was calculated on samples of pig brain contaminated with a Brazilian isolate of SuHV-1. The technique had a detection limit of 10-0,5 TCID50/50µL. PCR was able to distinguish nonspecific reactions due to primer dimers (thus avoiding false positives) and get faster results.

8436 downloads

10.

I Diretriz sobre o consumo de gorduras e saúde cardiovascular
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Santos, R.D.

; Gagliardi, A.C.M.

; Xavier, H.T.

; Magnoni, C.D.

; Cassani, R .

; Lottenberg, A.M.P.

; Casella Filho, A.

; Araújo, D.B.

; Cesena, F.Y.

; Alves, R.J.

; Fenelon, G.

; Nishioka, S.A.D.

; Faludi, A.A.

; Geloneze, B.

; Scherr, C.

; Kovacs, C.

; Tomazzela, C.

; Carla, C.

; Barrera-Arellano, D.

; Cintra, D.

; Quintão, E.

; Nakandakare, E.R.

; Fonseca, F.A.H.

; Pimentel, I.

; Santos, J.E.

; Bertolami, M.C.

; Rogero, M.

; Izar, M.C.

; Nakasato, M.

; Damasceno, N.R.T.

; Maranhão, R.

; Cassani, R.S.L.

; Perim, R.

; Ramos, S.

Arquivos Brasileiros de Cardiologia

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jan 2013, Volume 100 N. 1 Pages 1 - 40

Abstract: Pt | Text: Pt | PDF: Pt

290296 downloads

11.

Analysis of acylcarnitine profiles in umbilical cord blood and during the early neonatal period by electrospray ionization tandem mass spectrometry
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Vieira Neto, E.

; Fonseca, A.A.

; Almeida, R.F.

; Figueiredo, M.P.

; Porto, M.A.S.

; Ribeiro, M.G.

Brazilian Journal of Medical and Biological Research

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jun 2012, Volume 45 N. 6 Pages 546 - 556

Abstract: En | Text: En | PDF: En

Acylcarnitine profiling by electrospray ionization tandem mass spectrometry (ESI-MS/MS) is a potent tool for the diagnosis and screening of fatty acid oxidation and organic acid disorders. Few studies have analyzed free carnitine and acylcarnitines in dried blood spots (DBS) of umbilical cord blood (CB) and the postnatal changes in the concentrations of these analytes. We have investigated these metabolites in healthy exclusively breastfed neonates and examined possible effects of birth weight and gestational age. DBS of CB were collected from 162 adequate for gestational age neonates. Paired DBS of heel-prick blood were collected 4-8 days after birth from 106 of these neonates, the majority exclusively breastfed. Methanol extracts of DBS with deuterium-labeled internal standards were derivatized before analysis by ESI-MS/MS. Most of the analytes were measured using a full-scan method. The levels of the major long-chain acylcarnitines, palmitoylcarnitine, stearoylcarnitine, and oleoylcarnitine, increased by 27, 12, and 109%, respectively, in the first week of life. Free carnitine and acetylcarnitine had a modest increase: 8 and 11%, respectively. Propionylcarnitine presented a different behavior, decreasing 9% during the period. The correlations between birth weight or gestational age and the concentrations of the analytes in DBS were weak (r £ 0.20) or nonsignificant. Adaptation to breast milk as the sole source of nutrients can explain the increase of these metabolites along the early neonatal period. Acylcarnitine profiling in CB should have a role in the early detection of metabolic disorders in high-risk neonates.

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12.

PCR Multiplex para detecção dos principais herpesvírus neurológicos de ruminantes
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Fonseca Jr., A.A.

; Costa, E.A.

; Oliveira, T.S.

; Sales, E.B.

; Sales, M.L.

; Leite, R.C.

; Heneimann, M. B.

; Reis, J.K.P.

Arquivo Brasileiro de Medicina Veterinária e Zootecnia

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dec 2011, Volume 63 N. 6 Pages 1405 - 1413

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Desenvolveu-se uma PCR multiplex (mPCR) para diagnóstico diferencial de encefalite bovina causada por herpesvírus suíno 1 (SuHV-1), herpesvírus bovino 1 (BoHV-1), herpesvírus bovino 5 (BoHV-5) e herpesvírus ovino 2 (OvHV-2). Os iniciadores foram projetados após alinhamento de sequências disponíveis no banco de genomas (GenBank) e a reação foi padronizada levando-se em consideração a concentração dos reagentes e os tipos diferentes de DNA polimerase. Após determinação da especificidade e sensibilidade, 65 amostras de encéfalo de bovinos com síndrome neurológica foram submetidas à análise. A sensibilidade analítica para detecção de BoHV-1, BoHV-5 e SuHV-1 foi, respectivamente, 10(1,2) TCID50/50µL, 10(1,0) TCID50/50µL, 10(1,3) TCID50/50µL na reação multiplex. Das 65 amostras analisadas, 10 foram positivas para BoHV-5, uma para BoHV-1 e cinco para OvHV-2. A mPCR descrita neste trabalho mostrou-se uma técnica útil para o diagnóstico diferencial de enfermidades relacionadas ao sistema nervoso central de bovinos.

The aim of this study was to develop a multiplex PCR (mPCR) for the differential diagnosis of bovine encephalitis caused by the suid herpesvirus 1 (SuHV-1), bovine herpesvirus 1 (BoHV-1), bovine herpesvirus 5 (BoHV-5) and ovine herpesvirus 2 (OvHV -2). The primers were designed after alignment of sequences available in GenBank and the reaction was developed by taking into account the concentration of reagents and different types of DNA polymerase. After determining the specificity and sensitivity to PCR, 65 brain samples from cattle with neurological syndrome were submitted to the reaction. The analytical sensitivity for detection of BoHV-1, BoHV-5 and SuHV-1 was, respectively, 10(1,2) TCID50/50µL, 10(1,0) TCID50/50µL, 10(1,3) TCID50/50µL. Ten samples were positive for BoHV-5, one for BoHV-1, one for SuHV-1 and five for OvHV-2. The mPCR described here is a useful technique for the differential diagnosis of diseases related to the central nervous system of cattle.

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13.

Detecção dos genes de Staphylococcus aureus, enterotoxinas e de resistência à meticilina em leite
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Dias, N.L.

; Silva, D.C.B.

; Oliveira, D.C.B.S.

; Fonseca Junior, A.A.

; Sales, M.L.

; Silva, N.

Arquivo Brasileiro de Medicina Veterinária e Zootecnia

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dec 2011, Volume 63 N. 6 Pages 1547 - 1552

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Realizou-se a detecção do gene de Staphylococcus aureus, de enterotoxinas e de resistência à meticilina com extração de DNA feita diretamente de amostras de leite. Das 200 amostras estudadas, 145 (72,5%) amplificaram o gene femA, e estas foram analisadas quanto à presença dos genes sea, seb, sec e mecA. Os genes das enterotoxinas mais prevalentes foram: sea (60%), seb (37,9%) e sec (6,9%). Foram encontradas 18 amostras de leite (11,0 %) com S. aureus portadores do gene mecA. A detecção de S. aureus diretamente do leite, sem a necessidade de isolamento bacteriano e a caracterização do potencial enterotoxigênico, demonstra que a técnica de PCR é muito útil para estudos epidemiológicos das infecções estafilocócicas da glândula mamária. O alto percentual (72,5%) de amostras de leite positivas para a presença do gene femA sugere que S. aureus constitui um dos principais agentes causadores de infecções intramamárias na microrregião de Sete Lagoas-MG e que seu potencial enterotoxigênico e presença do gene mecA, que identifica o S. aureus resistente à meticlina, representa um risco potencial à saúde pública.

This work was performed to detect the Staphylococcus aureus gene, enterotoxins resistance to methicillin with the extraction of DNA directly from milk samples. Of the 200 samples studied 145 (72.5%) amplified the femA gene, which were analyzed regarding the presence of sea, seb, sec and mecA genes. The most prevalent enterotoxins genes were: sea (60%), seb (37.9%) and sec (6.9%). 18 milk samples (11 %) had S. aureus carrying the mecA gene. The detection of S. aureus directly from the milk, with no need for bacterial isolation and the characterization of the enterotoxigenic potential demonstrate that the PCR technique is very useful for epidemiological studies of staphylococcal infections of the mammary gland. The high percentage (72.5%) of positive milk samples for the presence of the femA gene suggests that S. aureus constitutes of the main agents which cause intramammary infections in the micro region of Sete Lagoas-MG and that its enterotoxigenic potential and the presence of the mecA gene, which identifies the S. aureus resistant to methicillin, represent a potential risk to public health.

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14.

PCR duplex para diferenciação de amostras vacinais e selvagens do vírus da doença de Aujeszky
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Fonseca Jr, A.A.

; Dias, N.L.

; Leite, R.C.

; Heinemann, M.B.

; Reis, J.K.P.

Arquivo Brasileiro de Medicina Veterinária e Zootecnia

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oct 2010, Volume 62 N. 5 Pages 1259 - 1262

Abstract: Pt | Text: Pt | PDF: Pt

2116 downloads Cited 1 time in SciELO

15.

Glutathione and the redox control system trypanothione/trypanothione reductase are involved in the protection of Leishmania spp. against nitrosothiol-induced cytotoxicity
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Romão, P.R.T.

; Tovar, J.

; Fonseca, S.G.

; Moraes, R.H.

; Cruz, A.K.

; Hothersall, J.S.

; Noronha-Dutra, A.A.

; Ferreira, S.H.

; Cunha, F.Q.

Brazilian Journal of Medical and Biological Research

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mar 2006, Volume 39 N. 3 Pages 355 - 363

Abstract: En | Text: En | PDF: En

Glutathione is the major intracellular antioxidant thiol protecting mammalian cells against oxidative stress induced by oxygen- and nitrogen-derived reactive species. In trypanosomes and leishmanias, trypanothione plays a central role in parasite protection against mammalian host defence systems by recycling trypanothione disulphide by the enzyme trypanothione reductase. Although Kinetoplastida parasites lack glutathione reductase, they maintain significant levels of glutathione. The aim of this study was to use Leishmania donovani trypanothione reductase gene mutant clones and different Leishmania species to examine the role of these two individual thiol systems in the protection mechanism against S-nitroso-N-acetyl-D,L-penicillamine (SNAP), a nitrogen-derived reactive species donor. We found that the resistance to SNAP of different species of Leishmania was inversely correlated with their glutathione concentration but not with their total low-molecular weight thiol content (about 0.18 nmol/10(7) parasites, regardless Leishmania species). The glutathione concentration in L. amazonensis, L. donovani, L. major, and L. braziliensis were 0.12, 0.10, 0.08, and 0.04 nmol/10(7) parasites, respectively. L. amazonensis, that have a higher level of glutathione, were less susceptible to SNAP (30 and 100 µM). The IC50 values of SNAP determined to L. amazonensis, L. donovani, L. major, and L. braziliensis were 207.8, 188.5, 160.9, and 83 µM, respectively. We also observed that L. donovani mutants carrying only one trypanothione reductase allele had a decreased capacity to survive (~40%) in the presence of SNAP (30-150 µM). In conclusion, the present data suggest that both antioxidant systems, glutathione and trypanothione/trypanothione reductase, participate in protection of Leishmania against the toxic effect of nitrogen-derived reactive species.

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