Efeito do extrato etanólico de própolis verde sobre a produção de anticorpos após imunização contra parvovírus canino (CPV) e coronavírus canino (CCoV)

Este estudo foi realizado para avaliar se extrato etanólico de própolis verde (EEP) pode interferir na produção de anticorpos específicos após imunização contra parvovírus (CPV) e coronavírus canino (CCoV). Camundongos foram vacinados com CPV e CCoV (0.75, 1.5 e 3 x 106 TCID50) com ou sem 400 μg/dose de EEP. Vinte e um dias após a terceira dose foi mensurado IgG sérica. A coadministração de EEP aumentou significativamente os níveis de IgG específica para o CPV em animais inoculados com a maior concentração do antígeno, e não teve influência sobre os níveis de anticorpos para CCoV. Os resultados indicam que o EEP tem ação imunomoduladora intimamente dependente do tipo e concentração do antígeno utilizado, sendo capaz de aumentar os níveis de anticorpos contra CPV.This study was designed to evaluate whether an ethanolic extract of green propolis (EEP) can interfere with production of specific antibodies after immunization against parvovirus (CPV) and canine coronavirus (CCoV). Mice were vaccinated with CPV and CCoV (0.75, 1.5 and 3 x 106 TCID50) with or without 400 μg/dose of the EEP. Twenty one days after the third dose was measured serum IgG. The co-administration of the EEP significantly enhanced serum specific IgG responses to CPV in animals inoculated with the highest concentration of the antigen, and had no influence on levels of antibodies to CCoV. The results indicate that the EEP has immunomodulatory action closely dependent on the type and concentration of antigen used, being able to increase the levels of antibodies to CPV

Initial Results of the International Efforts in Screening New Agents against Candida auris

This article belongs to the Special Issue Biology, Immunology, Epidemiology, and Therapy of Fungal Infections: A Themed Issue Dedicated to Professor David A. Stevens.Background: Candida auris is an emergent fungal pathogen and a global concern, mostly due to its resistance to many currently available antifungal drugs. Objective: Thus, in response to this challenge, we evaluated the in vitro activity of potential new drugs, diphenyl diselenide (PhSe)2 and nikkomycin Z (nikZ), alone and in association with currently available antifungals (azoles, echinocandins, and polyenes) against Candida auris. Methods: Clinical isolates of C. auris were tested in vitro. (PhSe)2 and nikZ activities were tested alone and in combination with amphotericin B, fluconazole, or the echinocandins, micafungin and caspofungin. Results: (PhSe)2 alone was unable to inhibit C. auris, and antagonism or indifferent effects were observed in the combination of this compound with the antifungals tested. NikZ appeared not active alone either, but frequently acted cooperatively with conventional antifungals. Conclusion: Our data show that (PhSe)2 appears to not have a good potential to be a candidate in the development of new drugs to treat C. auris, but that nikZ is worthy of further study.his work was accomplished with support from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brazil, within the scope of the Capes-PrInt Program—Financing Code 001info:eu-repo/semantics/publishedVersio

Expressão transitória da glicoproteína S1 recombinante do vírus da bronquite infecciosa em células de mamíferos

The infection with infectious bronchitis virus (IBV) has been the cause of significant economic losses in the poultry world. The IBV has several genotypes and serotypes that differ widely in their pathogenicity. Some strains have a worldwide distribution. Among the proteins that make up the IBV, the S glycoprotein (subdivided into S1 and S2) stands out for being highly glycosylated. The S1 subunit is the major inducer of neutralizing antibodies and protective immune response. It’s responsible for adsorption of the virus to receptors on host cells, to determine the serotype of IBV, and for being a major determinant of cell tropism. Due to the complexity of the processes that occurs during the synthesis of the glycoprotein S1, the expression system in mammalian cell has characteristics such as post-translational modifications, like glycosylation, which makes it more attractive for the synthesis of this protein. The present study aimed to clone and express transiently the synthetic gene S1 glycoprotein of IBV produced from consensus sequences of brazilian and international samples. It had been used HEK 293 (Human embrionary kidney) and HeLa (Human cervical cancer). The gene was cloned into mammalian expression vector and transfected in cell culture. After 96 hours the supernatant cell and the lysed cell were collected. Expression and purification of recombinant S1 glycoprotein (rS1-IBV) were verified using the technique of SDS-PAGE and subsequently characterized by Western blotting using hyperimmune serum anti-IBV. It had been detected rS1-IBV from the cell supernatant. There was no detection of the recombinant protein from cell lysed of both cell. The study proved that it is possible to express recombinant S1 glycoprotein in mammalian cells.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESA infecção pelo vírus da bronquite infecciosa (IBV) tem sido causa de importantes perdas econômicas na avicultura mundial. O IBV possui diversos genótipos e sorotipos que diferem amplamente em sua patogenicidade, e destes, algumas cepas possuem distribuição mundial. Dentre as proteínas que compõem o IBV a glicoproteína S (subdividida em S1 e S2) destaca-se por ser altamente glicosilada. A subunidade S1 é a principal indutora de anticorpos neutralizantes e resposta imune protetora, além de ser responsável pela adsorção do vírus aos receptores das células hospedeiras, por determinar o sorotipo de IBV, e por ser um dos principais determinantes do tropismo celular. Devido à complexidade dos processos que ocorrem durante a síntese da glicoproteína S1, o sistema de expressão em células de mamíferos é o que detém características, como as modificações pós-traducionais incluindo glicosilação, que o torna indicado para a síntese desta proteína. O presente estudo teve como objetivo clonar e expressar de forma transitória o gene sintético da glicoproteína do S1 do IBV elaborado a partir do consenso das sequências de amostras de campo brasileiras e internacionais, em células HEK 293 (Human embrionary kidney) e HeLa (Human cervical cancer). O gene foi clonado no vetor de expressão em células de mamíferos e transfectado nos cultivos celulares. Após 96 horas foi coletado o sobrenadante das células e o lisado celular. A expressão e purificação da glicoproteína S1 recombinante (rS1-IBV) foram verificadas a partir da técnica de SDS-PAGE e posteriormente caracterizada através do Western blotting utilizando soro hiperimune anti-IBV. Houve a detecção da rS1- IBV a partir do sobrenadante das células. Não houve detecção da proteína recombinante a partir do lisado celular de ambas as células. O estudo demonstrou que é possível expressar a glicoproteína S1 recombinante em células de mamíferos

Avian IgY antibodies: characteristics and applications in immunodiagnostic

Immunoglobulin Y (IgY) is the major antibody isotype in birds, reptiles, amphibia, and lungfish, playing a similar biological role as mammal IgG. Due to its phylogenetic distance, immune diversification and presence in the egg yolk, IgY provide a number of advantages in immunodiagnostic compared to IgG from mammals. Moreover, IgY production is in agreement with international efforts to reduce, refine and if possible, to replace animals in experimentation, contributing substantially in favor of animal welfare. This article presents an overview about structural and functional features, production and applications of IgY in immunodiagnostic, as well as the advantages of chicken antibodies use

PCR ESPÉCIE-ESPECÍFICO PARA O DIAGNÓSTICO DE ESPOROTRICOSE CAUSADA POR S. BRASILIENSIS

Despite thousands of sporotrichosis cases related to the zoonotic transmission of Sporothrix brasiliensis have been described, the diagnostic gold standard is still the classical culturing methods. The mycological culture results are available afterseven to 30 days of incubation. Since an early diagnosis contributes to improving thetreatment and to the spread control of this mycosis, studies evaluating faster diagnosticmethods are needed. Therefore, we aimed to evaluate the species-specific PCR for thesporotrichosis diagnosis caused by S. brasiliensis, using samples collected with a noninvasive technique. Seventy-four swabs from feline (n=64), canine (n=5), and human(n=5) suspect sporotrichosis cases were included. All samples were submitted to classicalmethods for diagnosis and to S. brasiliensis-specific PCR. Using mycological culture asthe gold standard, the diagnosis of S. brasiliensis-caused infection was confirmed in 69%(51/74) of the cases. PCR was positive in 30 out of these 51 cases, showing 59%sensitivity, 100% specificity, 72% accuracy, 100% positive predictive value, and 52%negative predictive value. Consequently, more studies are needed to elucidate theinterference factors that culminated with the high rate of false-negative results and thenoptimize this molecular test for the accurate diagnosis of infections caused by S.brasiliensis.Apesar de milhares de casos de esporotricose relacionados a transmissão zoonótica por Sporothrix brasiliensis venham sendo descritos, o diagnóstico padrão-ouro ainda é a partir do método clássico de cultura. O resultado da cultura micológica demora de sete a trinta dias para ficar disponível. Considerando que o diagnóstico precoce contribuiria para um tratamento prematuro e com isso, com o controle na disseminação desta micose, estudos avaliando um diagnóstico precoce são necessários. Portanto, o objetivo do presente estudo foi avaliar o PCR espécie-específico para o diagnóstico da esporotricose causada por S. brasiliensis, utilizando amostras coletas por uma técnica não invasiva. Foram incluídas setenta e quatro amostras de casos suspeitos de esporotricose (swabs) oriundos de felinos (n=64), caninos (n=5) e humanos (n=5). Todas as amostras foram submetidas ao método clássico de diagnóstico e ao PCR específico para S. brasiliensis. Utilizando a cultura micológica como padrão-ouro, o diagnóstico da infecção causada por S. brasiliensis foi confirmado em 69% (51/74) dos casos. O PCR foi positivo em 30 dos 51 casos, demonstrando 59% de sensibilidade, 100% de especificidade, 72% de acurácia, 100% de valor preditivo positivo e 52% de valor preditivo negativo. Consequentemente, mais estudos são necessários para elucidar os fatores de transferência que culminaram com a alta taxa de resultados falsos negativos e então otimizar este teste molecular para a acurácia diagnóstica para infecções causadas por S. brasiliensis

Supplemental intravenous n-3 fatty acids and n-3 fatty acid status and outcome in critically ill elderly patients in the ICU receiving enteral nutrition

Background & aims: N-3 fatty acids (FA) may have benefits in ICU patients. The aims were to identify whether FA status is altered in critical illness and to evaluate the effect of supplemental intravenous n-3 FA on plasma FA status and clinical outcome in ICU patients receiving enteral nutrition.Methods: Enterally fed patients (n ¼ 49; 60e80 years) were recruited in the first 48 h of ICU admission. Fifteen patients received n-3 FA emulsion (0.2 g/kg) over 6 h for 3 consecutive days, and 34 patients did not (control). Samples were collected before supplementation, and 24 and 72 h after the third infusion. Nineteen healthy elderly subjects were also studied; they gave a single blood sample. FA were measured in plasma phosphatidylcholine (PC).Results: Critically ill patients had altered plasma PC FA compared with healthy elderly subjects. Surviving ICU patients had higher levels of docosahexaenoic acid and total n-3 FA and a lower ratio of n-6:n-3 FA in plasma PC than non-survivors. Infusion of n-3 FA increased eicosapentaenoic, docosahexaenoic and total n-3 FA, and decreased arachidonic and total n-6 FA and n-6:n-3 FA and arachidonic:eicosapentaenoic acid ratios. Gas exchange was enhanced 72 h after the third n-3 FA infusion (p ¼ 0.001).Conclusions: Critically ill patients may have altered plasma FA profiles. A higher total n-3 FA and docosahexaenoic acid content in plasma PC is associated with survival and improved gas exchange

Treatment of Human Sporotrichosis Caused by <i>Sporothrix brasiliensis</i>

We describe the successful treatment of a series of 30 zoonotic sporotrichosis cases from southern Brazil. Sporothrix brasiliensis was the species genotypically identified in all 25 confirmed cases. Five other cases were classified as probable, without laboratory confirmation, but with clinical and epidemiological data of cat-transmitted sporotrichosis. Two isolates were sequenced by translation elongation factor-1 alpha (EF1α) loci in order to compare their sequences, and both of them showed distinct genotypes from S. brasiliensis strains from other Brazilian states. Itraconazole (ITZ) or potassium iodide (KI) were the first choice treatment in 28 and 2 cases, respectively. Microdilution assay showed a wild-type profile of S. brasiliensis isolates to ITZ. However, a lack of clinical response occurred in 42% of cases, especially those treated with ITZ 100 mg/day, and treatment needed modifications, by either increased doses or antifungal combinations. Clinical cure required a mean of 187 days of treatment, which was dependent on the clinical form of the disease and age of patients. Therapy, including dosages and durations, for cutaneous forms of sporotrichosis requires re-evaluation, since cases caused by S. brasiliensis may influence treatment efficacy