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1.
Viruses ; 14(11)2022 10 31.
Artigo em Inglês | MEDLINE | ID: mdl-36366521

RESUMO

Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is responsible for causing Coronavirus Disease-2019 (COVID-19), a heterogeneous clinical condition that manifests varying symptom severity according to the demographic profile of the studied population. While many studies have focused on the spread of COVID-19 in large urban centers in Brazil, few have evaluated medium or small cities in the Northeast region. The aims of this study were: (i) to identify risk factors for mortality from SARS-CoV-2 infection, (ii) to evaluate the gene expression patterns of key immune response pathways using nasopharyngeal swabs of COVID-19 patients, and (iii) to identify the circulating SARS-CoV-2 variants in the residents of a medium-sized city in Northeast Brazil. A total of 783 patients infected with SARS-CoV-2 between May 2020 and August 2021 were included in this study. Clinical-epidemiological data from patients who died and those who survived were compared. Patients were also retrospectively divided into three groups based on disease severity: asymptomatic, mild, and moderate/severe. Samples were added to a qPCR array for analyses of 84 genes involved with immune response pathways and sequenced using the Oxford Nanopore MinION technology. Having pre-existing comorbidity; being male; having cardiovascular disease, diabetes, and/or chronic obstructive pulmonary disease; and PCR cycle threshold (Ct) values under 22 were identified as risk factors for mortality. Analysis of the expression profiles of inflammatory pathway genes showed that the greater the infection severity, the greater the activation of inflammatory pathways, triggering the cytokine storm and downregulating anti-inflammatory pathways. Viral genome analysis revealed the circulation of multiple lineages, such as B.1, B.1.1.28, Alpha, and Gamma, suggesting that multiple introduction events had occurred over time. This study's findings help identify the specific strains and increase our understanding of the true state of local health. In addition, our data demonstrate that epidemiological and genomic surveillance together can help formulate public health strategies to guide governmental actions.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , Masculino , Feminino , SARS-CoV-2/genética , COVID-19/epidemiologia , Estudos Retrospectivos , Brasil/epidemiologia
2.
Br J Nutr ; 123(9): 1003-1012, 2020 05 14.
Artigo em Inglês | MEDLINE | ID: mdl-31964426

RESUMO

A child's diet contains nutrients and other substances that influence intestinal health. The present study aimed to evaluate the relations between complementary feeding, intestinal barrier function and environmental enteropathy (EE) in infants. Data from 233 children were obtained from the Brazilian site of the Etiology, Risk Factors, and Interactions of Enteric Infections and Malnutrition and the Consequences for Child Health and Development Project cohort study. Habitual dietary intake from complementary feeding was estimated using seven 24-h dietary recalls, from 9 to 15 months of age. Intestinal barrier function was assessed using the lactulose-mannitol test (L-M), and EE was determined as a composite measure using faecal biomarkers concentrations - α-1-antitrypsin, myeloperoxidase (MPO) and neopterin (NEO) at 15 months of age. The nutrient adequacies explored the associations between dietary intake and the intestinal biomarkers. Children showed adequate nutrient intakes (with the exception of fibre), impaired intestinal barrier function and intestinal inflammation. There was a negative correlation between energy adequacy and L-M (ρ = -0·19, P < 0·05) and between folate adequacy and NEO concentrations (ρ = -0·21, P < 0·01). In addition, there was a positive correlation between thiamine adequacy and MPO concentration (ρ = 0·22, P < 0·01) and between Ca adequacy and NEO concentration (ρ = 0·23; P < 0·01). Multiple linear regression models showed that energy intakes were inversely associated with intestinal barrier function (ß = -0·19, P = 0·02), and fibre intake was inversely associated with the EE scores (ß = -0·20, P = 0·04). Findings suggest that dietary intake from complementary feeding is associated with decreased intestinal barrier function and EE in children.


Assuntos
Dieta/normas , Enterite/etiologia , Fenômenos Fisiológicos da Nutrição do Lactente , Intestinos/fisiologia , Brasil/epidemiologia , Aleitamento Materno , Estudos de Coortes , Enterite/epidemiologia , Feminino , Humanos , Lactente , Masculino , Estado Nutricional
4.
Scand J Immunol ; 87(6): e12668, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29701883

RESUMO

Although the semi-invariant natural killer T cells (iNKT) are a small subpopulation of cells in the peripheral blood, they are presumed to play a role in early stages of infection against various pathogens, including protozoa. This work investigates the activation status and cytokine profile of iNKT cells during human Leishmania infantum and Leishmania braziliensis infection. We studied iNKT cells in patients with symptomatic active visceral leishmaniasis (AVL) (n = 8), patients with symptomatic active cutaneous leishmaniasis (ACL) (n = 13), negative endemic controls (NEC) (n = 6) and non-endemic controls (NonEC) (n = 6), with and without total Leishmania antigen stimulus (TLA). The number of iNKT cells in the peripheral blood of patients with ACL and AVL unaltered in relation to control groups. Moreover, the iNKT cells from ACL showed a hyperactivation profile compared to patients with AVL. Additionally, TLA induced IFN-gamma production in iNKT cells from patients with ACL, while in iNKT of patients with AVL, TLA induced a decrease in this cytokine. Higher IL-17 and IL-10 production by iNKT cells from patients with ACL were also observed compared to all other groups. There were no changes in iNKT IL-10-producing cells in AVL after TLA stimulation. However, TLA induced increase in IL-10 in iNKT cells in patients with ACL. These findings suggest that, although iNKT cells showed distinct profiles in patients with ACL and AVL, they play a dual role in immune modulation in both Leishmania infections.


Assuntos
Plasticidade Celular/imunologia , Leishmania braziliensis/imunologia , Leishmania infantum/imunologia , Leishmaniose Cutânea/imunologia , Leishmaniose Visceral/imunologia , Células T Matadoras Naturais/imunologia , Adulto , Antígenos de Protozoários/imunologia , Feminino , Humanos , Interferon gama/imunologia , Interleucina-10/metabolismo , Interleucina-17/metabolismo , Leishmaniose Cutânea/parasitologia , Leishmaniose Visceral/parasitologia , Ativação Linfocitária/imunologia , Contagem de Linfócitos , Masculino , Adulto Jovem
5.
Braz J Med Biol Res ; 49(10): e5294, 2016 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-27580004

RESUMO

Determination of left ventricular ejection fraction (LVEF) using in vivo imaging is the cardiac functional parameter most frequently employed in preclinical research. However, there is considerable conflict regarding the effects of anesthetic agents on LVEF. This study aimed at assessing the effects of various anesthetic agents on LVEF in hamsters using transthoracic echocardiography. Twelve female hamsters were submitted to echocardiography imaging separated by 1-week intervals under the following conditions: 1) conscious animals, 2) animals anesthetized with isoflurane (inhaled ISO, 3 L/min), 3) animals anesthetized with thiopental (TP, 50 mg/kg, intraperitoneal), and 4) animals anesthetized with 100 mg/kg ketamine plus 10 mg/kg xylazine injected intramuscularly (K/X). LVEF obtained under the effect of anesthetics (ISO=62.2±3.1%, TP=66.2±2.7% and K/X=75.8±1.6%) was significantly lower than that obtained in conscious animals (87.5±1.7%, P<0.0001). The K/X combination elicited significantly higher LVEF values compared to ISO (P<0.001) and TP (P<0.05). K/X was associated with a lower dispersion of individual LVEF values compared to the other anesthetics. Under K/X, the left ventricular end diastolic diameter (LVdD) was increased (0.60±0.01 cm) compared to conscious animals (0.41±0.02 cm), ISO (0.51±0.02 cm), and TP (0.55±0.01 cm), P<0.0001. The heart rate observed with K/X was significantly lower than in the remaining conditions. These results indicate that the K/X combination may be the best anesthetic option for the in vivo assessment of cardiac systolic function in hamsters, being associated with a lower LVEF reduction compared to the other agents and showing values closer to those of conscious animals with a lower dispersion of results.


Assuntos
Anestésicos/farmacologia , Volume Sistólico/efeitos dos fármacos , Função Ventricular Esquerda/efeitos dos fármacos , Animais , Combinação de Medicamentos , Ecocardiografia/métodos , Feminino , Frequência Cardíaca/efeitos dos fármacos , Ventrículos do Coração/diagnóstico por imagem , Ventrículos do Coração/efeitos dos fármacos , Isoflurano/farmacologia , Ketamina/farmacologia , Mesocricetus , Valores de Referência , Sístole/efeitos dos fármacos , Tiopental/farmacologia , Fatores de Tempo , Xilazina/farmacologia
6.
Genet Mol Res ; 14(4): 15754-68, 2015 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-26634543

RESUMO

Microbiological evaluation is one of the most important parameters for analyzing the viability of an oyster farming system, which addresses public health and ecological concerns. Here, the microbiological quality of the oyster Crassostrea rhizophorae cultivated in a monitored environment and from natural beds in Bahia, northeastern Brazil, was determined. Bacterial diversity in oysters was measured by polymerase chain reaction-denaturing gradient gel electrophoresis. Sequence analysis revealed that most bacterial species showed similarity with uncultured or unidentified bacteria from environmental samples, and were clustered into the phylum Proteobacteria. Diverse bacteria from cultivated (monitored) oyster samples were grouped in the same cluster with a high similarity index (above 79%). Microbiological analyses revealed that these oysters did not contain pathogens. These results reflect the natural balance of the microbial communities essential to the maintenance of health and in inhibiting pathogen colonization in the oyster. On the other hand, bacterial diversity of samples from native stocks in extractive areas displayed a similarity index varying between 55 and 77%, and all samples were clustered separately from each other and from the cluster of samples derived from the cultivation area. Microbiological analyses showed that oysters from the extractive area were not fit for human consumption. This reflected a different composition of the microbial community in this area, probably resulting from anthropic impact. Our study also demonstrated that low temperatures and high rainfall limits the bacterial concentration in tropical oysters. This is the first study analyzing the total bacterial community profiles of the oyster C. rhizophorae.


Assuntos
Agricultura , Bactérias/classificação , Biodiversidade , Crassostrea/microbiologia , Microbiota , Animais , Bactérias/genética , Brasil , Microbiologia Ambiental , Metagenoma , Metagenômica , Filogenia
7.
Genet Mol Res ; 14(3): 8658-62, 2015 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-26345797

RESUMO

The objective of this study was to verify whether Toxoplasma gondii is excreted in the milk of naturally infected sheep. In order to accomplish this, 275 lactating ewes were used; these were bred extensively in 17 estates distributed across nine cities. Polymerase chain reaction amplification was used to detect T. gondii DNA in milk samples, and the indirect immunofluorescence test was employed for the detection of anti-T. gondii IgG antibodies in the sera, with a cut-off value of 1:64. It was possible to verify the presence of the parasite DNA in 6.5% (18/275) of the studied animals. Anti-T. gondii antibodies were present in 41.5% of the animals studied (114/275). There was no correlation between parasite excretion in milk and the presence of IgG in 38.9% of the studied animals (7/18). The high seropositivity and the presence of parasite DNA in the milk led to the conclusion that T. gondii infection is present in the sheep population in southern and southwestern Bahia, and that there is a risk of the human population becoming infected due to the consumption of raw, in natura milk.


Assuntos
DNA de Protozoário/isolamento & purificação , Leite/parasitologia , Doenças dos Ovinos/parasitologia , Toxoplasma/genética , Toxoplasmose Animal/parasitologia , Animais , DNA de Protozoário/genética , Feminino , Prevalência , Ovinos/parasitologia , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/metabolismo , Carneiro Doméstico/parasitologia , Toxoplasmose Animal/epidemiologia , Toxoplasmose Animal/metabolismo
8.
Genet Mol Res ; 14(2): 4658-65, 2015 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-25966240

RESUMO

The aim of this study was to detect evidence of Toxoplasma gondii using polymerase chain reaction (PCR)-based techniques in oysters (Crassostrea rhizophorae) obtained from the southern coastal region of Bahia, Brazil. A total of 624 oysters were collected, and the gills and digestive glands were dissected. Each tissue sample was separated into pools containing tissues (of the same type) from three animals, leading to a total of 416 experimental samples for analysis (208 samples each from the gills and digestive glands). Molecular analysis using PCR-based detection of the T. gondii AF 146527 repetitive fragment yielded negative results for all samples. However, when nested-PCR was used for detection of the T. gondii SAG-1 gene, 17 samples were positive, with the gills being the tissue with maximal detection of the parasite. These positive results were confirmed by sample sequencing. It is therefore suggested that C. rhizophorae oysters are capable of filtering and retaining T. gondii oocysts in their tissue. This represents a risk to public health because they are traditionally ingested in natura.


Assuntos
DNA de Protozoário/análise , Ostreidae/parasitologia , Toxoplasma/genética , Animais , Reação em Cadeia da Polimerase
9.
Parasitol Int ; 63(3): 567-70, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24631791

RESUMO

Recent studies have demonstrated that strains of Toxoplasma gondii in Brazil are frequently different from those detected in other countries, thus making an accurate phylogenetic analysis difficult. The aim of this study was to genetically characterize T. gondii samples from sheep raised in southern Bahia and intended for human consumption, by means of PCR-RFLP and sequencing techniques. Experimental samples were obtained from 200 sheep brains purchased at butcher's shops in Itabuna, Bahia, Brazil. In total, three samples (#54, #124 and #127) were T. gondii-positive. The application of multilocus PCR-RFLP using ten molecular markers (SAG1, SAG2, SAG3, BTUB, c22-8, PK1, GRA6, L358, c-29-2 and Apico) revealed a single genotype common to all samples of this study, which differed from any other published T. gondii genotypes. An atypical allele was detected in the L358 genetic marker; this has not previously been shown in any other South American T. gondii isolates. Phylogenetic analysis on the sequences from multilocus PCR sequencing revealed that these three samples were classified into the same lineage. Extensive indel regions were detected in the Apico genetic marker. Together, our findings revealed a new Brazilian T. gondii genotype. Further research should be conducted to enrich the database of Brazilian T. gondii genotypes from different regions. This will make it possible to understand the phylogenetic relationship between isolates.


Assuntos
Parasitologia de Alimentos , Doenças dos Ovinos/parasitologia , Toxoplasma/genética , Toxoplasma/isolamento & purificação , Toxoplasmose Animal/parasitologia , Animais , Brasil , DNA de Protozoário/genética , Genótipo , Humanos , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Fragmento de Restrição , Análise de Sequência de DNA/veterinária , Ovinos , Toxoplasma/classificação
10.
Arq. bras. med. vet. zootec ; Arq. bras. med. vet. zootec. (Online);65(4): 946-954, Aug. 2013. ilus, graf, tab
Artigo em Português | LILACS | ID: lil-684446

RESUMO

As células-tronco mesenquimais (CTMs) diferenciam-se em várias linhagens e têm potencial de utilização na medicina regenerativa. As CTMs podem ser isoladas de vários tecidos de animais adultos. O objetivo deste estudo foi o isolamento das CTMs do tecido adiposo de cães, seu cultivo e diferenciação. Foram coletadas amostras de tecido adiposo subcutâneo de cinco cães. As CTMs foram isoladas, obtendo-se 146.803 (±49.533) células/g, cultivadas e diferenciadas em osteoblastos, adipócitos e condrócitos. Avaliaram-se a cinética do crescimento, a morfologia e a viabilidade celular. A caracterização citoquímica comprovou a natureza mesenquimal das células isoladas. O cultivo foi iniciado com 20.000 células/mL, verificando-se crescimento rápido até 72 horas (220.000 células/mL), fase exponencial entre 72 e 192 horas (455.000 células/mL), seguida de platô por saturação da densidade com 240 horas (355.000 células/mL). A viabilidade celular variou entre 96 e 100%. As CTMs em cultivo são fibroblásticas, fusiformes, com citoplasma basofílico e núcleo esférico. O comprimento médio das células variou entre 80,85 e 98,36µm, a largura média entre 17,40 e 28,79µm e o diâmetro médio do núcleo entre 15,46 e 17,74µm.


The applications of mesenchymal stem cells (MSCs) are becoming increasingly more promising for regenerative medicine and tissue engineering fields. MSCs can be isolated from adult animals from a variety of tissues, such as the adipose. This study focused on the isolation, culture and differentiation of MSCs from canine adipose tissue. Samples of subcutaneous adipose tissue from five dogs were collected. These cells were isolated, cultured and differentiated into osteoblasts, adipocytes and chondrocytes. We obtained 146,803 (±49,533) cells/g. Growth kinetics and viability studies were conducted during cell culture and the evaluation of cell differentiation was successfully performed by cytochemistry. The cell cultures were initiated with 20,000 MSCs/ml. Rapid growth was observed at 72 hours (220,000 cells/ml), the exponential phase between 72 and 192 hours (455,000 cells/ml) and saturation at 240 hours (355,000 cells/ml). The cellular viability ranged from 96 to 100%. MSCs in culture are fibroblastic cells, fusiform with basophilic cytoplasm and spherical nucleus. The length and width means of the cells and nuclear diameter ranged from 80.85-98.36µm, 17.40-28.79µm and 15.46-17.74µm respectively.


Assuntos
Animais , Biologia Celular , Citoplasma , Tecido Adiposo/anatomia & histologia , Cães/classificação
11.
Arq. bras. med. vet. zootec ; 65(4): 946-954, ago. 2013. ilus, graf, tab
Artigo em Português | VETINDEX | ID: vti-9800

RESUMO

As células-tronco mesenquimais (CTMs) diferenciam-se em várias linhagens e têm potencial de utilização na medicina regenerativa. As CTMs podem ser isoladas de vários tecidos de animais adultos. O objetivo deste estudo foi o isolamento das CTMs do tecido adiposo de cães, seu cultivo e diferenciação. Foram coletadas amostras de tecido adiposo subcutâneo de cinco cães. As CTMs foram isoladas, obtendo-se 146.803 (±49.533) células/g, cultivadas e diferenciadas em osteoblastos, adipócitos e condrócitos. Avaliaram-se a cinética do crescimento, a morfologia e a viabilidade celular. A caracterização citoquímica comprovou a natureza mesenquimal das células isoladas. O cultivo foi iniciado com 20.000 células/mL, verificando-se crescimento rápido até 72 horas (220.000 células/mL), fase exponencial entre 72 e 192 horas (455.000 células/mL), seguida de platô por saturação da densidade com 240 horas (355.000 células/mL). A viabilidade celular variou entre 96 e 100%. As CTMs em cultivo são fibroblásticas, fusiformes, com citoplasma basofílico e núcleo esférico. O comprimento médio das células variou entre 80,85 e 98,36µm, a largura média entre 17,40 e 28,79µm e o diâmetro médio do núcleo entre 15,46 e 17,74µm.(AU)


The applications of mesenchymal stem cells (MSCs) are becoming increasingly more promising for regenerative medicine and tissue engineering fields. MSCs can be isolated from adult animals from a variety of tissues, such as the adipose. This study focused on the isolation, culture and differentiation of MSCs from canine adipose tissue. Samples of subcutaneous adipose tissue from five dogs were collected. These cells were isolated, cultured and differentiated into osteoblasts, adipocytes and chondrocytes. We obtained 146,803 (±49,533) cells/g. Growth kinetics and viability studies were conducted during cell culture and the evaluation of cell differentiation was successfully performed by cytochemistry. The cell cultures were initiated with 20,000 MSCs/ml. Rapid growth was observed at 72 hours (220,000 cells/ml), the exponential phase between 72 and 192 hours (455,000 cells/ml) and saturation at 240 hours (355,000 cells/ml). The cellular viability ranged from 96 to 100%. MSCs in culture are fibroblastic cells, fusiform with basophilic cytoplasm and spherical nucleus. The length and width means of the cells and nuclear diameter ranged from 80.85-98.36µm, 17.40-28.79µm and 15.46-17.74µm respectively.(AU)


Assuntos
Animais , Tecido Adiposo/anatomia & histologia , Citoplasma , Biologia Celular , Cães/classificação
12.
Genet Mol Res ; 12(3): 2611-7, 2013 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-23546981

RESUMO

Changes in intestinal microbial flora during a 4-week period of Salmonella enterica serovar Enteritidis colonization in resistant mice (latent carrier animals) were evaluated using a culture independent method involving denaturing gradient gel electrophoresis. The contents of the ileocecal portion of the intestines produced 26 bands. Fifty-seven percent of the bands were expressed in more than 80% of the samples. Forty percent of the bands present in the negative control were common to all samples, and 60% differed from those obtained 12 h and 1, 5, 10, and 28 days post-inoculation (PI). A dendrogram distinguished the negative control as the external group, and 2 clusters were formed with 76% similarity, separating the 12-h PI and 3-day PI time points from the others. These groupings were also revealed through multivariate analysis in a principal component analysis and the Venn diagram. The production of interferon γ 12 h and 3 days PI may explain this brief imbalance in microbiota that was quickly reversed in the subsequent days. These findings demonstrate that S. enterica serovar Enteritidis can colonize the gut and persist in balance with the microbiota of resistant hosts.


Assuntos
Ceco/microbiologia , DNA Bacteriano/análise , Íleo/microbiologia , Microbiota , Infecções por Salmonella/microbiologia , Animais , Eletroforese em Gel de Gradiente Desnaturante , Camundongos , Camundongos Endogâmicos C57BL , RNA Ribossômico 16S/genética , Salmonella enteritidis/genética
13.
Vet Parasitol ; 189(2-4): 153-61, 2012 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-22677134

RESUMO

This study genetically Toxoplasma gondii isolates obtained from pigs intended for human consumption in northeastern Brazil; multilocus PCR-RFLP and sequencing techniques were utilized. Bioassays were conducted using the brain and tongue of 20 pig heads purchased at butcher shops in the city of Ilheus, Bahia, Brazil. Overall, 11 T. gondii isolates designated TgPgBr06-16 were identified. Application of multilocus PCR-RFLP with seven molecular markers (SAG1, SAG2, SAG3, BTUB, C22-8, PK1 and Apico) identified six different genotypes. Isolates TgPgBr 06, 08, 11, 12, 14 and 15 were indistinguishable by this technique, forming a single genotype; the remaining isolates were characterized as distinct genotypes. However, when five genetic markers (SAG1, SAG2, SAG3, BTUB and c22-8) were employed in multilocus PCR-sequencing, all eleven strains of T. gondii were shown to be different. All isolates differed from Type I, II and III clonal genotypes using both genotyping techniques. These results demonstrate that the multilocus PCR-RFLP assay underestimated the true diversity of the T. gondii population in this study. Thus, DNA sequencing is the preferred technique to infer the genetic diversity and population structure of T. gondii strains from Brazil. Moreover, it is necessary to develop new molecular markers to group and characterize atypical T. gondii isolates from South America.


Assuntos
Doenças dos Suínos/parasitologia , Toxoplasma/genética , Toxoplasmose Animal/parasitologia , Animais , Sequência de Bases , Brasil/epidemiologia , DNA de Protozoário/genética , Marcadores Genéticos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , Polimorfismo Genético , Polimorfismo de Fragmento de Restrição , Suínos , Doenças dos Suínos/epidemiologia , Toxoplasmose Animal/epidemiologia
14.
Genet Mol Res ; 10(4): 2578-88, 2011 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-22033939

RESUMO

Quantification of Salmonella in asymptomatic carrier animals can be used to assess microbial risk and monitor the level of contamination in domestic animals used for food production. We examined the sensitivity, specificity and accuracy of real-time qPCR, without pre-enrichment or selective enrichment stages, for the quantification of S. enterica serovar Enteritidis in resistant mice, as a model of asymptomatic carrier animal. The results were compared with those obtained by traditional bacteriological culture methods, the gold standard test. Two hundred and forty-three samples, including spleen, liver, mesenteric lymph nodes, portions of intestine, intestinal content of the ileocecal portion, and feces, were collected from a group of 27 C57BL/6 mice, that had been intragastrically inoculated with high doses of S. enterica serovar Enteritidis. The real-time qPCR assay presented a consistent linearity of the standard curve (r(2) = 0.999), with very low differences between melting temperatures, and low coefficients of variation in intra- (< 1%) and interassay (< 2%) comparisons. The primers were highly specific; there was no amplification with other Salmonella serovars or with DNA from uninfected tissues and feces from mice. The detection limit of the technique was defined as 32 copies of S. enterica serovar Enteritidis. A sensitivity of 90%, a specificity of 77% and an accuracy of 79% were obtained. The higher sensitivity of PCR was reflected in a kappa coefficient of 0.41, showing moderate agreement between tests. We conclude that real-time qPCR is a good alternative for diagnostic scanning in asymptomatic carrier animals, due to its high sensitivity and rapidity.


Assuntos
DNA Bacteriano/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Infecções por Salmonella , Salmonella enteritidis , Animais , Técnicas Bacteriológicas/métodos , Camundongos , Especificidade de Órgãos , Infecções por Salmonella/diagnóstico , Infecções por Salmonella/genética , Infecções por Salmonella/microbiologia , Salmonella enteritidis/genética , Salmonella enteritidis/crescimento & desenvolvimento , Salmonella enteritidis/isolamento & purificação , Sensibilidade e Especificidade
15.
Zoonoses Public Health ; 57(7-8): e26-32, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19968856

RESUMO

Species of tegu (Tupinambis) are the largest lizards in South America. Large numbers of these lizards are hunted; there is a vigorous trade in their skins and the meat is consumed by rural and native peoples. The animals are also bred in captivity, an economic activity for rural populations which can help in the animals' conservation. Faecal samples from 30 captive-born tegus were analysed for the presence of Salmonella in two separate samplings. In the first analysis, samples from 26 animals (87%) yielded Salmonella enterica of which 23% were of Rubislaw serotype; 20% Carrau and Agona serotypes; 7% Infantis and Saint-Paul serotypes; 3% Panama and Brandenburg serotypes; 10% were S. enterica subsp. enterica and 7% were rough form. In the second analysis, four tegus (13%) which had been negative in the first sampling were positive, thus, 100% of the animals studied carried the bacterium. Antibiotic susceptibility showed resistance to sulfonamide in 82% of the isolates, streptomycin in 64%, tetracycline in 6% and Chloramphenicol in 20%. Two animals carried strains of the same serotype with different patterns of antibiotic susceptibility. Although it is well known that reptiles are a significant source of Salmonella, to our knowledge, its prevalence in tegu has not been studied previously.


Assuntos
Antibacterianos/farmacologia , Lagartos/microbiologia , Salmonelose Animal/epidemiologia , Salmonella/efeitos dos fármacos , Salmonella/genética , Animais , DNA Bacteriano/análise , Farmacorresistência Bacteriana Múltipla , Fezes/microbiologia , Testes de Sensibilidade Microbiana , Panamá/epidemiologia , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/veterinária , Prevalência , Salmonella/classificação , Salmonella/isolamento & purificação , Salmonelose Animal/tratamento farmacológico , Salmonelose Animal/microbiologia , Sorotipagem/veterinária
16.
Braz J Med Biol Res ; 42(8): 744-9, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19649401

RESUMO

Amiodarone-induced thyroid dysfunction (AITD) is a common complication of amiodarone therapy and its prevalence varies according to iodine intake, subclinical thyroid disorders and the definition of AITD. There is no consensus about the frequency of screening for this condition. We evaluated 121 patients on chronic regular intake of amiodarone (mean intake = 248.5 +/- 89 mg; duration of treatment = 5.3 +/- 3.9 years, range = 0.57-17 years) and with stable baseline cardiac condition. Those with no AITD were followed up for a median period of 3.2 years (range: 0.6-6.7) and the incidence rate of AITD, defined by clinical and laboratorial findings as proposed by international guidelines, was obtained (62.8 per 1000 patients/year). We applied the Cox proportional hazard model to adjust for potential confounding factors and used sensitivity analysis to identify the best screening time for follow-up. We detected thyroid dysfunction in 59 (48.7%) of the 121 patients, amiodarone-induced hypothyroidism in 50 (41.3%) and hyperthyroidism in 9 (7.5%). Compared with patients without AITD, there was no difference regarding dosage or duration of therapy, heart rhythm disorder or baseline cardiac condition. During the follow-up of the 62 patients without AITD at baseline evaluation, 11 developed AITD (interquartile range, IR: 62.8 (95%CI: 31.3-112.3) cases per 1000 patients/year), 9 of them with hypothyroidism - IR: 11.4 (95%CI: 1.38-41.2), and 2 hyperthyroidism - IR: 51.3 (95%CI: 23.4-97.5). Age, gender, dose, and duration of treatment were not significant after adjustment. During the first 6 months of follow-up the incidence rate for AITD was 39.3 (9.2-61.9) cases per 1000 patients/year. These data show that AITD is quite common, and support the need for screening at 6-month intervals, unless clinical follow-up dictates otherwise or further information regarding the prognosis of untreated subclinical AITD is available.


Assuntos
Amiodarona/efeitos adversos , Antiarrítmicos/efeitos adversos , Hipertireoidismo/induzido quimicamente , Hipotireoidismo/induzido quimicamente , Idoso , Amiodarona/uso terapêutico , Antiarrítmicos/uso terapêutico , Arritmias Cardíacas/tratamento farmacológico , Seguimentos , Humanos , Hipertireoidismo/diagnóstico , Hipotireoidismo/diagnóstico , Masculino , Pessoa de Meia-Idade , Fatores de Tempo
17.
Rev. bras. pesqui. méd. biol ; Braz. j. med. biol. res;42(8): 744-749, Aug. 2009. graf, tab
Artigo em Inglês | LILACS | ID: lil-520781

RESUMO

Amiodarone-induced thyroid dysfunction (AITD) is a common complication of amiodarone therapy and its prevalence varies according to iodine intake, subclinical thyroid disorders and the definition of AITD. There is no consensus about the frequency of screening for this condition. We evaluated 121 patients on chronic regular intake of amiodarone (mean intake = 248.5 ± 89 mg; duration of treatment = 5.3 ± 3.9 years, range = 0.57-17 years) and with stable baseline cardiac condition. Those with noAITD were followed up for a median period of 3.2 years (range: 0.6-6.7) and the incidence rate of AITD, defined by clinical and laboratorial findings as proposed by international guidelines, was obtained (62.8 per 1000 patients/year). We applied the Coxproportional hazard model to adjust for potential confounding factors and used sensitivity analysis to identify the best screening time for follow-up. We detected thyroid dysfunction in 59 (48.7%) of the 121 patients, amiodarone-induced hypothyroidism in50 (41.3%) and hyperthyroidism in 9 (7.5%). Compared with patients without AITD, there was no difference regarding dosage or duration of therapy, heart rhythm disorder or baseline cardiac condition. During the follow-up of the 62 patients without AITD at baseline evaluation, 11 developed AITD (interquartile range, IR: 62.8 (95%CI: 31.3-112.3) cases per 1000 patients/year), 9 of them with hypothyroidism - IR: 11.4 (95%CI: 1.38-41.2), and 2 hyperthyroidism - IR: 51.3 (95%CI: 23.4-97.5). Age, gender,dose, and duration of treatment were not significant after adjustment. During the first 6 months of follow-up the incidence rate for AITD was 39.3 (9.2-61.9) cases per 1000 patients/year. These data show that AITD is quite common, and support the need for screening at 6-month intervals, unless clinical follow-up dictates otherwise or further information regarding the prognosis of untreated subclinical AITD is available.


Assuntos
Idoso , Humanos , Masculino , Pessoa de Meia-Idade , Amiodarona/efeitos adversos , Antiarrítmicos/efeitos adversos , Hipertireoidismo/induzido quimicamente , Hipotireoidismo/induzido quimicamente , Amiodarona/uso terapêutico , Antiarrítmicos/uso terapêutico , Arritmias Cardíacas/tratamento farmacológico , Seguimentos , Hipertireoidismo/diagnóstico , Hipotireoidismo/diagnóstico , Fatores de Tempo
18.
Genet Mol Res ; 8(1): 375-88, 2009 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-19440973

RESUMO

Landfarm soil is used to bioremediate oil wastes from petrochemical industries. We developed a simplified protocol for microbial DNA extraction of tropical landfarm soil using only direct lysis of macerated material. Two samples of tropical landfarm soil from a Brazilian refinery were analyzed by this protocol (one consisted of crude oil-contaminated soil; the other was continuously enriched for nine months with petroleum). The soil samples were lysed by maceration with liquid nitrogen, eliminating the need for detergents, organic solvents and enzymatic cell lysis. Then, the DNA from the lysed soil sample was extracted using phenol-chloroform-isoamyl alcohol or guanidium isothiocyanate, giving high DNA yields (more than 1 micro g DNA/g soil) from both soil types. This protocol compared favorably with an established method of DNA template preparation that included mechanical, chemical and enzymatic treatment for cell lysis. The efficiency of this extraction protocol was confirmed by polymerase chain reaction amplification of the 16S rRNA gene, denaturing gradient gel electrophoresis and cloning assays. Fifty-one different clones were obtained; their sequences were classified into at least seven different phyla of the Eubacteria group (Proteobacteria - alpha, gamma and delta, Chloroflexi, Actinobacteria, Acidobac teria, Planctomycetes, Bacteroidetes, and Firmicutes). Forty percent of the sequences could not be classified into these phyla, demonstrating the genetic diversity of this microbial community. Only eight isolates had sequences similar to known sequences of 16S rRNA of cultivable organisms or of known environmental isolates and therefore could be identified to the genus level. This method of DNA extraction is a useful tool for analysis of the bacteria responsible for petroleum degradation in contaminated environments.


Assuntos
Biodiversidade , DNA Bacteriano/isolamento & purificação , DNA Ribossômico/isolamento & purificação , Petróleo , RNA Ribossômico 16S/genética , Microbiologia do Solo , Poluentes do Solo/análise , Biodegradação Ambiental , Microbiologia Ambiental , Reação em Cadeia da Polimerase , Clima Tropical
19.
Transplant Proc ; 40(3): 875-8, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18455041

RESUMO

Chagas disease (American trypanosomiasis) is caused by the protozoan parasite Trypanosoma cruzi. Chagas disease following solid-organ transplantation has occurred in Latin America. This report presents the occurrence of Chagas disease despite negative serological tests in both the donor and the recipient, as well as the effectiveness of treatment. A 21-year-old woman from the state of Sao Paulo (Brazil) underwent cadaveric donor liver transplantation in November 2005, due to cirrhosis of autoimmune etiology. Ten months after liver transplantation, she developed signs and symptoms of congestive heart failure (New York Heart Association functional class IV). The echocardiogram, which was normal preoperatively, showed dilated cardiac chambers, depressed left ventricular systolic function (ejection fraction = 35%) and moderate pulmonary hypertension. Clinical investigation discarded ischemic heart disease and autoimmune and other causes for heart failure. Immuno fluorescence (immunoglobulin M and immunoglobulin G) and hemagglutination tests for T cruzi were positive, and abundant T cruzi amastigotes were readily identified in myocardial biopsy specimens. Treatment with benznidazole for 2 months yielded an excellent clinical response. At the moment of submission, the patient remains in functional class I. This case highlighted that more appropriate screening for T cruzi infection is mandatory in potential donors and recipients of solid-organ transplants in regions where Chagas disease is prevalent. Moreover, it stressed that this diagnosis should always be considered in recipients who develop cardiac complications, since negative serological tests do not completely discard the possibility of disease transmission and since good results can be achieved with prompt trypanocidal therapy.


Assuntos
Cardiomiopatia Chagásica/diagnóstico , Transplante de Fígado/efeitos adversos , Complicações Pós-Operatórias/parasitologia , Trypanosoma cruzi/isolamento & purificação , Adulto , Animais , Cardiomiopatia Chagásica/tratamento farmacológico , Ecocardiografia , Evolução Fatal , Coração/parasitologia , Humanos , Masculino , Nitroimidazóis/uso terapêutico , Transplante de Pâncreas , Tripanossomicidas/uso terapêutico , Disfunção Ventricular Esquerda
20.
J Investig Allergol Clin Immunol ; 16(4): 247-52, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16889282

RESUMO

BACKGROUND: There is some evidence suggesting a link between paracetamol exposure and atopy in both adults and children. OBJECTIVE: We aimed to investigate whether further epidemiological support for a link between paracetamol intake and allergy could be found in a population of Mexican children. METHODS: In a cross-sectional study design, we applied the ISAAC questionnaire to 3493 children aged 6 to 7 years old. Two analyses were performed: (1) children were classified as cases if they had wheezing, rhinitis, or eczema at any time from their neonatal period up until they reached the age of 6 to 7 years, or as controls if they had never experienced these conditions, and (2) children were classified as cases if they had wheezing, rhinitis, or eczema in the 12 months prior to the study. Paracetamol intake was considered positive if it frequently occurred during the first year of life (first analysis) or in the last 12 months (second analysis). RESULTS: Paracetamol intake in the first year of life was significantly associated with an increased risk of ev having wheezing (adjusted odds ratio [OR], 1.69; 95% confidence interval [CI], 1.23 to 2.34) and rhinitis (adjusted OR, 1.37; 95% CI, 1.20 to 1.59) but not eczema (adjusted OR, 1.45; 95% CI, 0.91 to 2.32). Frequent paracetamol intake in the last year increased the risk of wheezing (OR, 3.3; 95% CI, 1.54 to 7.18), rhinitis (OR, 1.61; 95% CI, 1.33 to 1.95), or eczema (OR, 1.82; 95% CI, 1.24 to 2.66). CONCLUSION: Frequent paracetamol exposure was associated with a significantly increased risk of wheezing and rhinitis and probably eczema in a Mexican population of children.


Assuntos
Acetaminofen/efeitos adversos , Dermatite Atópica/induzido quimicamente , Sons Respiratórios/etiologia , Rinite/induzido quimicamente , Acetaminofen/administração & dosagem , Analgésicos não Narcóticos/administração & dosagem , Analgésicos não Narcóticos/efeitos adversos , Antibacterianos/administração & dosagem , Antibacterianos/efeitos adversos , Estudos de Casos e Controles , Criança , Pré-Escolar , Dermatite Atópica/etiologia , Dermatite Atópica/metabolismo , Feminino , Humanos , Hipersensibilidade Imediata/induzido quimicamente , Hipersensibilidade Imediata/etiologia , Hipersensibilidade Imediata/metabolismo , Lactente , Recém-Nascido , Masculino , México , Estresse Oxidativo/efeitos dos fármacos , Rinite/etiologia , Rinite/metabolismo , Fatores de Risco , Inquéritos e Questionários
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