No description available.

Background and Objectives: The pathophysiology of post-operative pneumonia following lung resection is poorly understood despite it being the most common complication which may lead to death. The role of the acute inflammatory response following lung resection, in particular innate immune cells, was investigated and used to identify biomarkers for post-operative pneumonia. Comparison of inflammatory responses to resection undertaken by video-assisted thorascopic surgery (VATS) and thoracotomy was also evaluated. Methods: Patients undergoing lung resection for suspected bronchogenic carcinoma were recruited. Objective pre-defined criteria were used to diagnose pneumonia. Bronchoalveolar lavage (BAL) was conducted in the contra-lateral lung pre- and postoperatively to measure cellular composition and cytokines. Blood was sampled preoperatively and 6-, 24- and 48-hours post-operatively primarily to assess neutrophil phagocytic capacity, monocyte subsets, monocyte cytokine responses to lipopolysaccharide (LPS) stimulation and serum cytokine responses. Exhaled nitric oxide (eNO) was also measured at these time points. Patient groups were compared using paired or student t-tests together with ANOVA/ANCOVA modelling. The predictive strength of the biomarkers identified was tested. Results: 40 patients were recruited. 26 patients (65%) underwent major lung resection using VATS and 14 (35%) thoracotomy. There was a post-operative blood monocytosis (p<0.0005) with an absolute expansion of classical and intermediate monocytes (p=0.001) and a relative fall in non-classical monocytes (p<0.005). Post-operatively blood monocytes became more pro-inflammatory with an overall significant increase in IL-8 (p=0.034) and TNF-α (p=0.028) together with an increase in IL-6 (p=0.028) and IL-10 by 48 hours (p=0.010). VATS was associated with a smaller release of IL-10 only (p=0.011). There was a general trend towards post-operative reduction in neutrophil phagocytosis of zymosan (in suspension) on ANOVA modelling (p=0.047). Lung resection led to an increase in serum cytokines IL-6, IL-8 and IL-10 which peaked at 24hrs before falling (p<0.0005). ANOVA modelling confirmed significantly lower levels of serum cytokines in VATS patients compared with thoracotomy (p=0.026 for IL-6, p=0.018 for IL-8 p=0.047 for IL-10). No significant post-operative change was found for IL-1β, TNF-α and IL-12p70 (p>0.05). Bronchoalveolar lavage fluid (BALF) and blood samples demonstrated a relative post-operative leucocytosis due principally to neutrophilia. A relative blood lymphopenia and thrombocytopenia developed postoperatively (p<0.0005). VATS was associated with a lower fall in serum albumin (p=0.001). BALF from the non-operated lung became more pro-inflammatory immediately post-operatively with an increase in IL-6 (p<0.0005), IL-8 (p=0.017), IL- 10 (p=0.018) and IL-1β (p=0.002). eNO tended to fall post-operatively which reached significance at 48 hrs (p=0.029). 14 patients developed pneumonia. Pre-operatively, a blood neutrophil count above 5.04x109/L had a relative risk (RR) for pneumonia of 3.3 (95% confidence interval (CI95) 1.1-10.1), and a BAL cell count of greater than 1.04x105/ml had a RR of 3.4 (CI95 1.3-9.0), whilst LPS-stimulated monocyte secretion of IL-12 of less than 0.15 pg/ml/μg protein had a RR of 3.0 (CI95 1.2-7.3). At 24 hours post-operatively, LPS-stimulated release from monocytes of IL-10 greater than 1.99 pg/ml/μg protein (RR 4.1, CI95 1.3- 12.3) and IL-6 greater than 414 pg/ml/μg protein (RR 3.1, CI95 1.2-8.1) were predictive of pneumonia. Conclusion: Lung resection is associated with significant early pro- and antiinflammatory responses. VATS resection invoked significantly lower levels of serum cytokines and albumin changes compared with thoracotomy suggesting VATS lobectomy should be the surgical treatment strategy of choice for early stage lung cancer. No difference in neutrophil function or monocyte function was however observed between the surgical groups. Clinical benefits of this reduced inflammation need to be evaluated in a larger cohort of patients. Relative pre-operative leucocytosis in blood and BAL together with monocyte hyper-responsiveness in the early postoperative period is associated with the development of pneumonia. These findings warrant further investigation for their predictive power in accurately identifying postoperative pneumonia. Ultimately, they may be incorporated into a risk stratification model enabling targeted prophylactic or earlier therapeutic intervention.

O treinamento aeróbio moderado tem sido reconhecido como um importante estimulador do sistema imune, no entanto o efeito deste na infecção bacteriana não tem sido extensivamente estudado. Nosso objetivo foi avaliar se o exercício aeróbio moderado prévio à infecção por S. pneumoniae influencia a resposta inflamatória pulmonar. Camundongos BALB/C foram divididos em 4 grupos: Controle (animais sedentários; não infectados); S. pneumoniae (animais sedentários e posteriormente infectados); Exercício (animais treinados; não infectados); Exercício + S. pneumoniae (animais treinados e posteriormente infectados). Os animais foram submetidos a um programa de treinamento físico aeróbio durante 4 semanas, e 72 horas após a última sessão de exercício, os animais receberam instilação nasal de S. pneumoniae (linhagem M10) e foram avaliados 12 horas (fase aguda) ou 10 dias (fase tardia) após a instilação. Na fase aguda, o grupo S. pneumoniae apresentou um aumento de: resistência e elastância do sistema respiratório, número total de células, neutrófilos, linfócitos e macrófagos no lavado broncoalveolar (BAL), células polimorfonucleares no parênquima pulmonar e TNF-alfa e IL-1beta no homogenato pulmonar. O exercício físico atenuou significantemente esses parâmentros. Além disso, o exercício físico resultou em aumento da expressão de enzimas antioxidantes no pulmão (CuZnSOD and MnSOD). Na fase tardia, o grupo Exercício + S. pneumoniae apresentou redução no número total de células e macrófagos no BAL, células polimorfonucleares no parênquima pulmonar e IL-6 no homogenato pulmonar comparado ao grupo S. pneumoniae. Nossos resultados sugerem um efeito protetor do exercício aeróbio moderado contra a infecção bacteriana pulmonar. Esse efeito é provavelmente secundário ao efeito do exercício no balanço oxidante-antioxidante / Moderate aerobic exercise training has been recognized as an important stimulator of the immune system, but its effect on bacterial infection has not been extensively studied. Our aim was to determine whether moderate aerobic exercise training prior to S. pneumoniae infection influences pulmonary inflammatory responses. BALB/c mice were divided into 4 groups: Control (sedentary without infection); S. pneumoniae (sedentary with infection); Exercise (aerobic training without infection); Exercise + S. pneumoniae (aerobic training with infection). Animals underwent aerobic exercise training for 4 weeks. 72 h after last exercise training, animals received a challenge with S. pneumoniae (strain M10) and were evaluated either 12 h (acute phase) or 10 days (late phase) after instillation. In acute phase, S. pneumoniae group had an increase in respiratory system resistance and elastance; number of total cells, neutrophils, lymphocytes and macrophages in bronchoalveolar lavage fluid (BAL); polymorphonuclear cells in lung parenchyma; and levels of TNF-alfa and IL-1beta in lung homogenates. Exercise training significantly attenuated the increase in all of these parameters. In addition, exercise induced an increase in expression of antioxidant enzymes (CuZnSOD and MnSOD) in lungs. In late phase, Exercise + S. pneumoniae group exhibited a reduction in number of total cells and macrophages in BAL, in polymorphonuclear cells in lung parenchyma and in levels of IL-6 in lung homogenates compared to S. pneumoniae group. Our results suggest a protective effect of moderate exercise training against respiratory infection with S. pneumoniae. This effect is most likely secondary to an effect of exercise on oxidant-antioxidant balance

O treinamento aeróbio moderado tem sido reconhecido como um importante estimulador do sistema imune, no entanto o efeito deste na infecção bacteriana não tem sido extensivamente estudado. Nosso objetivo foi avaliar se o exercício aeróbio moderado prévio à infecção por S. pneumoniae influencia a resposta inflamatória pulmonar. Camundongos BALB/C foram divididos em 4 grupos: Controle (animais sedentários; não infectados); S. pneumoniae (animais sedentários e posteriormente infectados); Exercício (animais treinados; não infectados); Exercício + S. pneumoniae (animais treinados e posteriormente infectados). Os animais foram submetidos a um programa de treinamento físico aeróbio durante 4 semanas, e 72 horas após a última sessão de exercício, os animais receberam instilação nasal de S. pneumoniae (linhagem M10) e foram avaliados 12 horas (fase aguda) ou 10 dias (fase tardia) após a instilação. Na fase aguda, o grupo S. pneumoniae apresentou um aumento de: resistência e elastância do sistema respiratório, número total de células, neutrófilos, linfócitos e macrófagos no lavado broncoalveolar (BAL), células polimorfonucleares no parênquima pulmonar e TNF-alfa e IL-1beta no homogenato pulmonar. O exercício físico atenuou significantemente esses parâmentros. Além disso, o exercício físico resultou em aumento da expressão de enzimas antioxidantes no pulmão (CuZnSOD and MnSOD). Na fase tardia, o grupo Exercício + S. pneumoniae apresentou redução no número total de células e macrófagos no BAL, células polimorfonucleares no parênquima pulmonar e IL-6 no homogenato pulmonar comparado ao grupo S. pneumoniae. Nossos resultados sugerem um efeito protetor do exercício aeróbio moderado contra a infecção bacteriana pulmonar. Esse efeito é provavelmente secundário ao efeito do exercício no balanço oxidante-antioxidante / Moderate aerobic exercise training has been recognized as an important stimulator of the immune system, but its effect on bacterial infection has not been extensively studied. Our aim was to determine whether moderate aerobic exercise training prior to S. pneumoniae infection influences pulmonary inflammatory responses. BALB/c mice were divided into 4 groups: Control (sedentary without infection); S. pneumoniae (sedentary with infection); Exercise (aerobic training without infection); Exercise + S. pneumoniae (aerobic training with infection). Animals underwent aerobic exercise training for 4 weeks. 72 h after last exercise training, animals received a challenge with S. pneumoniae (strain M10) and were evaluated either 12 h (acute phase) or 10 days (late phase) after instillation. In acute phase, S. pneumoniae group had an increase in respiratory system resistance and elastance; number of total cells, neutrophils, lymphocytes and macrophages in bronchoalveolar lavage fluid (BAL); polymorphonuclear cells in lung parenchyma; and levels of TNF-alfa and IL-1beta in lung homogenates. Exercise training significantly attenuated the increase in all of these parameters. In addition, exercise induced an increase in expression of antioxidant enzymes (CuZnSOD and MnSOD) in lungs. In late phase, Exercise + S. pneumoniae group exhibited a reduction in number of total cells and macrophages in BAL, in polymorphonuclear cells in lung parenchyma and in levels of IL-6 in lung homogenates compared to S. pneumoniae group. Our results suggest a protective effect of moderate exercise training against respiratory infection with S. pneumoniae. This effect is most likely secondary to an effect of exercise on oxidant-antioxidant balance

Os complexos Mycobacterium chelonae M.abscessus e Mycobacterium fortuitum - M. peregrinum são compostos por espécies bacterianas de crescimento rápido e potencialmente patogênicas. Sua distribuição é ubíqua no ambiente, são resistentes a cloração da água e a sua replicação ocorre mesmo em condições de escassez de nutrientes. Estão envolvidos em casos de infecção pulmonar e extrapulmonar, e causam infecções em pacientes imunocomprometidos ou submetidos a rocedimentos cirúrgicos invasivos. Os objetivos do presente trabalho foram: confirmar através de testes fenotípicos e com as técnicas de PRA hsp65 e seqüenciamento do fragmento do rpoB, a identificação de micobactérias de crescimento rápido, incluídas nos complexos M. chelonae-M.abscessus e M. fortuitum-M.peregrinum. Foram incluídos no estudo os isolados provenientes de pacientes com dois ou mais isolamentos provenientes de sítio não estéril ou um isolamento de sítio estéril. O estudo de 38 isolados demonstrou que as provas fenotípicas disponíveis atualmente não permitem a identificação de todas as espécies de micobactérias de crescimento rápido já descritas na literatura. O PRA hsp65 possibilitou a identificação rápida e precisa de 63% das espécies de micobactérias e demonstrou um perfil compartilhado pelas espécies M. abscessus 2; M. bolletii 1 e M. massiliense 1. O seqüenciamento do gene rpoB confirmou a identificação das espécies citadas. Nossos resultados demonstraram que o PRA-hsp65 e o seqüenciamento do gene rpoB são ferramentas úteis para fornecer a identificação das espécies de micobactérias com mais acurácia. O uso dessas técnicas poderiam ser consideradas em laboratórios de referência para identificar Micobactérias de crescimento rápido uma vez que elas são patógenos emergentes implicados em surtos e isolados de pacientes em centros de referência para tratamento de tuberculose multirresistente. / Mycobacterium chelonae-M. abscessus and Mycobacterium fortuitum-M. peregrinum complexes are composed by bacterial species characterized by rapid grow and considered as potential pathogens. These microorganisms are ubiquitous in the environment, resistant to water treatment such as standard chlorination and are able to replicate even at poor nutrient conditions. They are related to lung and extralung infections in immune-compromised patients or those submitted to invasive surgical procedures. The aim of this study was to confirm the identification of Mycobacterium chelonae - M. abscessus and Mycobacterium fortuitum - M. peregrinum complexes, isolated from biological samples considering one or two repetitions if samples are originated from sterile or non-sterile site respectively. Phenotypic tests and molecular methods, PRA-hsp65 and rpoB gene sequencing, were applied to identify 38 strains previously isolated. Results demonstrated that available phenotypic tests did not allow the identification of all described fast growing Mycobacterium. The PRA of hsp65 gene confirmed the identification of 63% of the Mycobacterium studied, and demonstrated the band pattern shared by M. abscessus 2, M. bolletii and M. massiliensis. The rpoB gene sequencing confirmed the identification of the species cited. Our results demonstrated that PRA-hsp65 and rpoB gene sequencing are useful tools to provide a more accurate species identification of mycobacteria. The use of such techniques would be considered in reference laboratories to identify fast growing Mycobacterium species since they are considered emerging pathogens implicated in outbreaks and isolated from a patient in reference centers for treatment of multidrug resistant tuberculosis.

Os complexos Mycobacterium chelonae M.abscessus e Mycobacterium fortuitum - M. peregrinum são compostos por espécies bacterianas de crescimento rápido e potencialmente patogênicas. Sua distribuição é ubíqua no ambiente, são resistentes a cloração da água e a sua replicação ocorre mesmo em condições de escassez de nutrientes. Estão envolvidos em casos de infecção pulmonar e extrapulmonar, e causam infecções em pacientes imunocomprometidos ou submetidos a rocedimentos cirúrgicos invasivos. Os objetivos do presente trabalho foram: confirmar através de testes fenotípicos e com as técnicas de PRA hsp65 e seqüenciamento do fragmento do rpoB, a identificação de micobactérias de crescimento rápido, incluídas nos complexos M. chelonae-M.abscessus e M. fortuitum-M.peregrinum. Foram incluídos no estudo os isolados provenientes de pacientes com dois ou mais isolamentos provenientes de sítio não estéril ou um isolamento de sítio estéril. O estudo de 38 isolados demonstrou que as provas fenotípicas disponíveis atualmente não permitem a identificação de todas as espécies de micobactérias de crescimento rápido já descritas na literatura. O PRA hsp65 possibilitou a identificação rápida e precisa de 63% das espécies de micobactérias e demonstrou um perfil compartilhado pelas espécies M. abscessus 2; M. bolletii 1 e M. massiliense 1. O seqüenciamento do gene rpoB confirmou a identificação das espécies citadas. Nossos resultados demonstraram que o PRA-hsp65 e o seqüenciamento do gene rpoB são ferramentas úteis para fornecer a identificação das espécies de micobactérias com mais acurácia. O uso dessas técnicas poderiam ser consideradas em laboratórios de referência para identificar Micobactérias de crescimento rápido uma vez que elas são patógenos emergentes implicados em surtos e isolados de pacientes em centros de referência para tratamento de tuberculose multirresistente. / Mycobacterium chelonae-M. abscessus and Mycobacterium fortuitum-M. peregrinum complexes are composed by bacterial species characterized by rapid grow and considered as potential pathogens. These microorganisms are ubiquitous in the environment, resistant to water treatment such as standard chlorination and are able to replicate even at poor nutrient conditions. They are related to lung and extralung infections in immune-compromised patients or those submitted to invasive surgical procedures. The aim of this study was to confirm the identification of Mycobacterium chelonae - M. abscessus and Mycobacterium fortuitum - M. peregrinum complexes, isolated from biological samples considering one or two repetitions if samples are originated from sterile or non-sterile site respectively. Phenotypic tests and molecular methods, PRA-hsp65 and rpoB gene sequencing, were applied to identify 38 strains previously isolated. Results demonstrated that available phenotypic tests did not allow the identification of all described fast growing Mycobacterium. The PRA of hsp65 gene confirmed the identification of 63% of the Mycobacterium studied, and demonstrated the band pattern shared by M. abscessus 2, M. bolletii and M. massiliensis. The rpoB gene sequencing confirmed the identification of the species cited. Our results demonstrated that PRA-hsp65 and rpoB gene sequencing are useful tools to provide a more accurate species identification of mycobacteria. The use of such techniques would be considered in reference laboratories to identify fast growing Mycobacterium species since they are considered emerging pathogens implicated in outbreaks and isolated from a patient in reference centers for treatment of multidrug resistant tuberculosis.

No description available.

Pneumonia is one of the leading causes of death worldwide. The course of the disease is often highly dynamic with unforeseen critical deterioration within hours in a relevant proportion of patients. Besides antibiotic treatment, novel adjunctive therapies are under development. Their additive value needs to be explored in preclinical and clinical studies and corresponding therapy schedules require optimization prior to introduction into clinical practice. Biomathematical modeling of the underlying disease and therapy processes might be a useful aid to support these processes. We here propose a biomathematical model of murine immune response during infection with Streptococcus pneumoniae aiming at predicting the outcome of different treatment schedules. The model consists of a number of non-linear ordinary differential equations describing the dynamics and interactions of the pulmonal pneumococcal population and relevant cells of the innate immune response, namely alveolar- and inflammatory macrophages and neutrophils. The cytokines IL-6 and IL-10 and the chemokines CCL2, CXCL1 and CXCL5 are considered as major mediators of the immune response. We also model the invasion of peripheral blood monocytes, their differentiation into macrophages and bacterial penetration through the epithelial barrier causing blood stream infections. We impose therapy effects on this system by modelling antibiotic therapy and treatment with the novel C5a-inactivator NOX-D19. All equations are derived by translating known biological mechanisms into equations and assuming appropriate response kinetics. Unknown model parameters were determined by fitting the predictions of the model to time series data derived from mice experiments with close-meshed time series of state parameters. Parameter fittings resulted in a good agreement of model and data for the experimental scenarios. The model can be used to predict the performance of alternative schedules of combined antibiotic and NOX-D19 treatment. We conclude that we established a comprehensive biomathematical model of pneumococcal lung infection, immune response and barrier function in mice allowing simulations of new treatment schedules. We aim to validate the model on the basis of further experimental data. We also plan the inclusion of further novel therapy principles and the translation of the model to the human situation in the near future.

Aviäre pathogene E. coli (APEC) sind extraintestinale Pathogene, die beim Huhn systemische Infektionskrankheiten hervorrufen. Zur Identifizierung Gene, die an der Kolonisierung des Wirtes beteiligt sind, wurde ein Lungen-Infektionsmodell in 5 Wochen alten SPF Hühnern etabliert. In dem Modell wurden 1.800 mittels Signature-tagged-Mutagenese (STM) hergestellten Mutanten des APEC Stamms IMT5155 (O2:K1:H5; ST-Komplex 95) auf ihre Fähigkeit zur Kolonisierung getestet. Die Untersuchung führte zur Identifizierung Gene, einschließlich Adhäsin-, LPS- und Kapsel-bildenden Genen, sowie Genen mit putativer Funktion. Die STM-Analyse erlaubte zudem die Identifizierung eines zuvor nicht charakterisierten putativen Fimbrien-bildenden Adhäsins (Yqi). Das Genprodukt wurde vorläufig als ExPEC Adhäsin I (EA/I) bezeichnet. Eine Deletion des EA/I-Gens führte zu einer Reduzierung der Adhäsionsfähigkeit des Stammes IMT5155 in vitro und in vivo. Eine Komplementierung des EA/I-Gens in trans resultierte in einer Wiederherstellung des Adhäsions¬vermögens in vitro. Das EA/I-Protein (~39 kDa) wurde als Fusionsprotein in vitro exprimiert, und mittels SDS-PAGE und Western Blot nachgewiesen. Durch Überexpression des EA/I-Operons in dem Fimbrien-negativen E. coli-Stamm AAEC189 konnten mittels elektronenmikroskopischer Aufnahmen Fimbrien-bildende Strukturen auf der äußeren Membran dargestellt werden. Das Vorkommen des yqi in den untersuchten extraintestinal pathogenen E. coli (ExPEC), bei gleichzeitigem Fehlen in allen untersuchten intestinal pathogenen E. coli bestätigt die Bezeichnung ExPEC Adhäsin I. Die Prävalenz des EA/I-Gens war am stärksten assoziiert mit Stämmen der B2-Phylogenetische-Gruppe und des ST95-Komplexes des Multi-Lokus-Sequenz-Typisierungs (MLST)-Schemas. Sequenzanalysen ergaben zudem erste Hinweise auf eine positive Selektion des EA/I-Gens innerhalb dieses Komplexes. In der vorliegenden Arbeit gelang somit die Identifizierung und Charakterisierung des neuen ExPEC Adhäsin I. / The extraintestinal pathogen, avian pathogenic E. coli (APEC), known to cause systemic infections in chickens, is responsible for large economic losses in the poultry industry. To identify genes, involved adhesion and colonization, a lung colonization model of infection was established in 5-week old specific-pathogen free (SPF) chickens, and Signature-tagged mutagenesis (STM) was applied to this model by generating and screening a total of 1,800 mutants of an APEC strain IMT5155 (O2:K1:H5; ST complex 95). This led to the identification of new genes of interest, including adhesins, genes involved in capsule and LPS formation, and genes of putative function. Among the many genes identified was one coding for a novel APEC fimbrial adhesin (Yqi) not described for its role in APEC pathogenesis. Its gene product was temporarily designated ExPEC Adhesin I (EA/I). Deletion of the ExPEC adhesin I gene resulted in reduced colonization ability by APEC strain IMT5155 both in vitro and in vivo. Complementation of the adhesin gene restored its ability to colonize epithelial cells in vitro. The ExPEC adhesin I protein (~ 39 kDa) was expressed as a fusion protein in vitro as shown by SDS-PAGE and western blotting. Electron microscopy of an afimbriate strain E. coli AAEC189 over-expressed with the putative EA/I gene cluster revealed short fimbrial like appendages protruding out of the bacterial outer membrane. We observed that the adhesin coding gene yqi is prevalent among extraintestinal pathogenic E. coli (ExPEC) isolates and absent in all of the intestinal pathogenic E. coli strains tested, thereby validating the designation of the adhesin as ExPEC Adhesin I. In addition, prevalence of EA/I was most frequently associated with the E. coli phylogenetic group B2 and ST95 complex of the multi locus sequence typing (MLST) scheme, with evidence of a positive selection within this complex. This is the first report of the newly identified and functionally characterized ExPEC adhesin I.