Immunological responses and cytokine gene expression analysis to Cooperia punctata infections in resistant and susceptible Nelore cattle

Cellular and humoral immune response, as well as cytokine gene expression, was assessed in Nelore cattle with different degrees of resistance to Cooperia punctata natural infection. One hundred cattle (male, weaned, 11-12 months old), kept together on pasture, were evaluated. Faecal and blood samples were collected for parasitological and immunological assays. Based on nematode faecal egg counts (FEC) and worm burden, the seven most resistant and the eight most susceptible animals were selected. Tissue samples of the small intestine were collected for histological quantification of inflammatory cells and analysis of cytokine gene expression (IL-2, IL-4, IL-8, IL-1 2p35, IL-13, TNF-alpha, IFN-gamma, MCP-1, MCP-2, and MUC- 1) using real-time RT-PCR. Mucus samples were also collected for IgA levels determination. Serum IgG1 mean levels against C. punctata antigens were higher in the resistant group, but significant differences between groups were only observed 14 days after the beginning of the experiment against infective larvae (1-3) and 14 and 84 days against adult antigens. The resistant group also presented higher IgA levels against C. punctata (L3 and adult) antigens with significant difference 14 days after the beginning of the trial (P < 0.05). In the small-intestine mucosa, levels of IgA anti-L3 and anti-adult C. punctata were higher in the resistant group, compared with the susceptible group (P < 0.05). Gene expression of both T(H)2 cytokines (IL-4 and IL-13) in the resistant group and T(H)1 cytokines (IL-2, IL-1 2p35, IFN-gamma and MCP-1) in the susceptible group was up-regulated. Such results suggested that immune response to C. punctata was probably mediated by TH2 cytokines in the resistant group and by T(H)1 cytokines in the susceptible group. (C) 2008 Elsevier B.V. All rights reserved.

A morphometric study of the midgut in resistant and non-resistant Anticarsia gemmatalis (Hubner) (Lepidoptera: Noctuidae) larvae to its nucleopolyhedrovirus (AgMNPV)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Moxidectin-resistant nematodes in cattle in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Esterase profile in a pyrethroid-resistant Brazilian strain of the cattle tick Boophilus microplus (Acari, Ixodidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Highly corrosion resistant siloxane-polymethyl methacrylate hybrid coatings

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Genetic markers between Biomphalaria glabrata snails susceptible and resistant to Schistosoma mansoni infection

The analysis of the genetic variability related to susceptibility to Schistosoma mansoni infection in the vector of the genus Biomphalaria is important in terms of a better understanding of the epidemiology of schistosomiasis itself, the possible pathological implications of this interaction in vertebrate hosts, and the formulation of new strategies and approaches for disease control. In the present study, the genetic variability of B. glabrata strains found to be resistant or susceptible to S. mansoni infection was investigated using DNA amplification by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). The amplification products were analyzed on 8% polyacrylamide gel and stained with silver. We selected 10 primers, since they have previously been useful to detect polymorphism among B. glabrata and/or B. tenagophila. The results showed polymorphisms with 5 primers. Polymorphic bands observed only in the susceptible strain. The RAPD-PCR methodology represents an adequate approach for the analysis of genetic polymorphisms. The understanding of the genetic polymorphisms associated to resistance may contribute to the future identification of genomic sequences related to the resistance/susceptibility of Biomphalaria to the larval forms of S. mansoni and to the development of new strategies for the control of schistosomiasis.

Characterization of ndh gene of isoniazid resistant and susceptible Mycobacterium tuberculosis isolates from Brazil

Resistance in Mycobacterium tuberculosis to isoniazid (INH) is caused by mutations in the catalase-peroxidase gene (katG) , and within the inhA promoter and/or in structural gene. A small percentage (~ 10%) of INH-resistant strains do not present mutations in both of these loci. Other genes have been associated with INH resistance including the gene encoding for NADH dehydrogenase (ndh) . Here we report the detection of two ndh locus mutations (CGT to TGT change in codon 13 and GTG to GCG change in codon 18) by analyzing 23 INH-resistant and in none of 13 susceptible isolates from Brazilian tuberculosis patients. We also detected two isolates without a mutation in ndh, or any of the other INH resistance-associated loci examined, suggesting the existence of additional, as yet to be described, INH resistance mechanisms.

Fruit quality of 'Tahiti' lime (Citrus latifolia Tanak) grafted on twelve different rootstocks

No presente trabalho, avaliou-se a influência de doze porta-enxertos sobre a qualidade dos frutos da lima-ácida 'Tahiti' (Citrus latifolia Tanaka), clone 'IAC-5', amostrados em duas posições nas plantas, em experimento conduzido na Estação de Citricultura de Bebedouro, em um pomar de três anos. O espaçamento utilizado foi de 8.0 x 5.0m. Os porta-enxertos utilizados foram: citrangeiro 'Carrizo' (C. sinensis (L.) Osbeck x Poncirus trifoliata (L.) Raf.); os híbridos 'Cravo' x 'Swingle' (C. limonia Osbeck x P. trifoliata (L.) Raf. x C. paradisi Macf.) e 'Changsha' x 'English Small' (C. sunki Hort. ex Tan. x P. trifoliata Raf.); as tangerineiras 'Sun Chu Sha Kat' (C. reticulata Blanco) e 'Sunki' (C. sunki Hort. ex Tanaka); os limoeiros 'Cravo Limeira' e 'Cravo FCAV' (C. limonia Osbeck); o citrumeleiro 'Swingle' (P. trifoliata Raf. x C. paradisi Macf.), o tangeleiro 'Orlando' (C. reticulata Blanco x C. paradisi Macf.) e os trifoliateiros 'Rubidoux', 'FCAV' e 'Flying Dragon' (Poncirus trifoliata Raf.). Foi utilizado um delineamento em blocos casualizados, com doze tratamentos e seis repetições. Os distintos porta-enxertos induziram diferenças na qualidade dos frutos, entretanto todas as características de qualidade foram consideradas aceitáveis para a variedade, sendo bons substitutos para o limão 'Cravo'.

Nasopharyngeal colonization with methicillin-resistant staphylococcus aureus and mortality among patients in an intensive care unit

A colonização de nasofaringe por Staphylococcus aureus, resistente à meticilina (Methicillin-resistant S.aureus - MRSA), é comum em pacientes criticamente doentes, mas seu significado prognóstico não é inteiramente conhecido. Realizou-se estudo de coorte retrospectivo com 122 pacientes de uma unidade de terapia intensiva que realizaram triagem semanal para colonização por MRSA. Os desfechos de interesse foram: mortalidade geral e mortalidade por infecção. Diversas variáveis de exposição (gravidade, procedimentos, intercorrências e colonização nasofaríngea por MRSA) foram analisadas em modelos univariados e multivariados. Fatores significativamente associados à mortalidade geral ou por infecção foram: APACHE II e doença pulmonar. A colonização por MRSA não foi preditora de mortalidade geral (OR=1,02; IC95%=0,35-3; p=0,97) ou por infecção (OR=0,96; IC95%=0,33-2,89; p=0,96). Os resultados sugerem que, na ausência de fatores de gravidade, a colonização por MRSA não caracteriza pior prognóstico.

Case-case-control study of risk factors for nasopharyngeal colonization with methicillin-resistant Staphylococcus aureus in a medical-surgical intensive care unit

Nasopharyngeal colonization with methicillin-resistant Staphylococcus aureus (MRSA) often precedes the development of nosocomial infections. In order to identify risk factors for MRSA colonization, we conducted a case-case-control study, enrolling 122 patients admitted to a medical-surgical intensive care unit (ICU). All patients had been screened for nasopharyngeal colonization with S. aureus upon admission and weekly thereafter. Two case-control studies were performed, using as cases patients who acquired colonization with MRSA and methicillin-susceptible S. aureus (MSSA), respectively. For both studies, patients in whom colonization was not detected during ICU stay were selected as control subjects. Several potential risk factors were assessed in univariate and multivariable (logistic regression) analysis. MRSA and MSSA were recovered from nasopharyngeal samples from 27 and 10 patients, respectively. Independent risk factors for MRSA colonization were: length-of-stay in the ICU (Odds Ratio [OR]=1.12, 95%Confidence Interval[CI]=1.06-1.19, p<0.001) and use of ciprofloxacin (OR=5.05, 95%CI=1.38-21.90, p=0.015). The use of levofloxacin had a protective effect (OR=0.08, 95%CI=0.01-0.55, p=0.01). Colonization with MSSA was positively associated with central nervous system disease (OR=7.45, 95%CI=1.33-41.74, p=0.02) and negatively associated with age (OR=0.94, 95%CI=0.90-0.99, p=0.01). In conclusion, our study suggests a role for both cross-transmission and selective pressure of antimicrobials in the spread of MRSA.

Comparative ''in vitro'' study on the susceptibility and emergence of mutants resistant to danofloxacin and ciprofloxacin among Staphylococcus aureus isolated from bovine mastitis

The antimicrobial susceptibility of danofloxacin was compared in vitro with ciprofloxacin against mastitogenic Staphylococcus aureus. Danofloxacin was more active than ciprofloxacin, showing minimal inhibitory concentrations (MIC90), minimal bactericidal concentrations (MBC90) and MBC/MIC ratio lower than those of ciprofloxacin. Ciprofloxacin-resistant mutants occurred at higher frequencies (greater than or equal to 10(-6)) than for danofloxacin (10(-9)).

Crystallographic and pre-steady-state kinetics studies on binding of NADH to wild-type and isoniazid-resistant enoyl-ACP(CoA) reductase enzymes from Mycobacterium tuberculosis

An understanding of isoniazid (INH) drug resistance mechanism in Mycobacterium tuberculosis should provide significant insight for the development of newer anti-tubercular agents able to control INH-resistant tuberculosis (TB). The inhA-encoded 2-trans enoyl-acyl carrier protein reductase enzyme (InhA) has been shown through biochemical and genetic studies to be the primary target for INH. In agreement with these results, mutations in the inhA structural gene have been found in INH-resistant clinical isolates of M. tuberculosis, the causative agent of TB. In addition, the InhA mutants were shown to have higher dissociation constant values for NADH and lower values for the apparent first-order rate constant for INH inactivation as compared to wild-type InhA. Here, in trying to identify structural changes between wild-type and INH-resistant InhA enzymes, we have solved the crystal structures of wild-type and of S94A, I47T and I21V InhA proteins in complex with NADH to resolutions of, respectively, 2.3 angstrom, 2.2 angstrom, 2.0 angstrom, and 1.9 angstrom. The more prominent structural differences are located in, and appear to indirectly affect, the dinucleotide binding loop structure. Moreover, studies on pre-steady-state kinetics of NADH binding have been carried out. The results showed that the limiting rate constant values for NADH dissociation from the InhA-NADH binary complexes (k(off)) were eleven, five, and tenfold higher for, respectively, I21V, I47T and S94A INH-resistant mutants of InhA as compared to INH-sensitive wildtype InhA. Accordingly, these results are proposed to be able to account for the reduction in affinity for NADH for the INH-resistant InhA enzymes. (c) 2006 Elsevier Ltd. All rights reserved.

Study of pulmonary experimental paracoccidioidomycosis by analysis of bronchoalveolar lavage cells: Resistant vs. susceptible mice

Adult Swiss (susceptible) and BALB/c (non-susceptible) mice were inoculated by the intravenous route with 1 x 10(6) yeast cells of Paracoccidioides brasiliensis, strain 18. Immunologic parameters, histopathology and features of the bronchoalveolar lavage (BAL) were evaluated at week 2, 4, 8 and 16 post-infection. The pulmonary infection was progressive in Swiss mice and regressive in BALB/c mice. The numbers of total cells, lymphocytes and polymorphonuclear neutrophils increased in BAL, as well as the percentages of giant cells, and CD4 and CD8 positive cells. The ultrastructural study of BAL cells revealed a predominance of macrophages and a frequency of 13.2% of type II pneumocytes. As the infection progressed, the number of fungal cells and spreading macrophages, as well as the stimulated release of H2O2 by macrophages, increased. The animals exhibited an exacerbation of the humoral immune response and a depression of cellular immunity during the infection. There was a good correlation between the intensity and the pattern of the pulmonary histopathology and the cellular findings in the BAL. The present model reproduces some anatomoclinical patterns of the human disease and shows that BAL may be a useful tool in monitoring the pulmonary infection caused by P. brasiliensis.

Crystallographic studies on the binding of isonicotinyl-NAD adduct to wild-type and isoniazid resistant 2-trans-enoyl-ACP (CoA) reductase from Mycobacterium tuberculosis

The resumption of tuberculosis led to an increased need to understand the molecular mechanisms of drug action and drug resistance, which should provide significant insight into the development of newer compounds. Isoniazid (INH), the most prescribed drug to treat TB, inhibits an NADH-dependent enoyl-acyl carrier protein reductase (InhA) that provides precursors of mycolic acids, which are components of the mycobacterial cell wall. InhA is the major target of the mode of action of isoniazid. INH is a pro-drug that needs activation to form the inhibitory INH-NAD adduct. Missense mutations in the inhA structural gene have been identified in clinical isolates of Mycobacterium tuberculosis resistant to INH. To understand the mechanism of resistance to INH, we have solved the structure of two InhA mutants (121V and S94A), identified in INH-resistant clinical isolates, and compare them to INH-sensitive WT InhA structure in complex with the INH-NAD adduct. We also solved the structure of unliganded INH-resistant S94A protein, which is the first report on apo form of InhA. The salient features of these structures are discussed and should provide structural information to improve our understanding of the mechanism of action of, and resistance to, INH in M. tuberculosis. The unliganded structure of InhA allows identification of conformational changes upon ligand binding and should help structure-based drug design of more potent antimycobacterial agents. (c) 2007 Elsevier B.V. All rights reserved.