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In order to survive and replicate, Salmonella has evolved mechanisms to gain access to intestinal epithelial cells of the crypt. However, the impact of Salmonella Typhimurium on stem cells and progenitors, which are responsible for the ability of the intestinal epithelium to renew and protect itself, remains unclear. Given that intestinal organoids growth is sustained by stem cells and progenitors activity, we have used this model to document the effects of Salmonella Typhimurium infection on epithelial proliferation and differentiation, and compared it to an in vivo model of Salmonella infection in mice. Among gut segments, the caecum was preferentially targeted by Salmonella. Analysis of infected crypts and organoids demonstrated increased length and size, respectively. mRNA transcription profiles of infected crypts and organoids pointed to upregulated EGFR-dependent signals, associated with a decrease in secretory cell lineage differentiation. To conclude, we show that organoids are suited to mimic the impact of Salmonella on stem cells and progenitors cells, carrying a great potential to drastically reduce the use of animals for scientific studies on that topic. In both models, the EGFR pathway, crucial to stem cells and progenitors proliferation and differentiation, is dysregulated by Salmonella, suggesting that repeated infections might have consequences on crypt integrity and further oncogenesis. [ABSTRACT FROM AUTHOR]

A tunable ultrashort soliton pulse source reaching up to 4.8 µm is demonstrated based on a 2.8 µm femtosecond fiber laser coupled to a zirconium fluoride fiber amplifier followed by a small core indium fluoride fiber. This demonstration is extending by 300 nm the long wavelength limit previously reported with soliton self-frequency shift (SSFS) sources based on fluoride fibers. Our experimental and numerical investigation highlighted the spectral dynamics associated with the generation of highly redshifted pulses in the mid-infrared using SSFS enhanced by soliton fission. This study is intended at providing a better understanding of the potential and limitations of SSFS based tunable femtosecond fiber sources in the 3–5 µm spectral range. [ABSTRACT FROM AUTHOR]

Excessive lung inflammation and airway epithelial damage are hallmarks of human inflammatory lung diseases, such as cystic fibrosis (CF). Enhancement of innate immunity provides protection against pathogens while reducing lung-damaging inflammation. However, the mechanisms underlying innate immunity-mediated protection in the lung remain mysterious, in part because of the lack of appropriate animal models for these human diseases. TLR5 (Toll-like receptor 5) stimulation by its specific ligand, the bacterial protein flagellin, has been proposed to enhance protection against several respiratory infectious diseases, although other cellular events, such as calcium signaling, may also control the intensity of the innate immune response. Here, we investigated the molecular events prompted by stimulation with flagellin and its role in regulating innate immunity in the lung of the pig, which is anatomically and genetically more similar to humans than rodent models. We found that flagellin treatment modulated NF-kB signaling and intracellular calcium homeostasis in airway epithelial cells. Flagellin pretreatment reduced the NF-kB nuclear translocation and the expression of proinflammatory cytokines to a second flagellin stimulus as well as to Pseudomonas aeruginosa infection. Moreover, in vivo administration of flagellin decreased the severity of P. aeruginosa-induced pneumonia. Then we confirmed these beneficial effects of flagellin in a pathological model of CF by using ex vivo precision-cut lung slices from a CF pigz model. These results provide evidence that flagellin treatment contributes to a better regulation of the inflammatory response in inflammatory lung diseases such as CF. [ABSTRACT FROM AUTHOR]

The main features of lung infection and inflammation are a massive recruitment of neutrophils and the subsequent release of neutrophil serine proteases (NSPs). Anti-infectious and/or anti-inflammatory treatments must be tested on a suitable animal model. Mice models do not replicate several aspects of human lung disease. This is particularly true for cystic fibrosis (CF), which has led the scientific community to a search for new animal models. We have shown that mice are not appropriate for characterizing drugs targeting neutrophil-dependent inflammation and that pig neutrophils and their NSPs are similar to their human homologues. We induced acute neutrophilic inflammatory responses in pig lungs using Pseudomonas aeruginosa, an opportunistic respiratory pathogen. Blood samples, nasal swabs and bronchoalveolar lavage fluids (BALFs) were collected at 0, 3, 6 and 24 h post-insfection (p.i.) and biochemical parameters, serum and BAL cytokines, bacterial cultures and neutrophil activity were evaluated. The release of proinflammatory mediators, biochemical and hematological blood parameters, cell recruitment and bronchial reactivity, peaked at 6h p.i.. We also used synthetic substrates specific for human neutrophil proteases to show that the activity of pig NSPs in BALFs increased. These proteases were also detected at the surface of lung neutrophils using anti-human NSP antibodies. Pseudomonas aeruginosa-induced lung infection in pigs results in a neutrophilic response similar to that described for cystic fibrosis and ventilator-associated pneumonia in humans. Altogether, this indicates that the pig is an appropriate model for testing anti-infectious and/or anti-inflammatory drugs to combat adverse proteolytic effects of neutrophil in human lung diseases. [ABSTRACT FROM AUTHOR]

An aqueous marine-sulfated polysaccharide (MSP) extract, prepared from the green macroalga Ulva armoricana, was tested as an antibacterial compound against 42 bacterial strains and isolates found in livestock animals. Both Gram-positive and Gram-negative bacteria growths were affected. The most susceptible pathogens were Pasteurella multocida, Mannheimia haemolytica, Erysipelothrix rhusiopathiae, Staphylococcus aureus, and Streptococcus suis, with a minimum inhibitory concentration (MIC) ranging from 0.16 to 6.25 mg mL. Enterococcus cecorum, Streptococcus dysgalactiae, Corynebacterium, Trueperella pyogenes, and Bordetella bronchiseptica strains were also susceptive to MSP with a complete inhibition recorded at MIC values of 25 and 50 mg mL. The stimulation of the immune response mediators of the host's gut with the extract was evaluated using an in vitro system of differentiated porcine intestinal epithelial cells (IPEC-1). RT-qPCR analysis showed a significant increase of mRNA expression of cytokines such as IL1α, IL1β, L6, IL8, TNFα, and TGFβ as well as the chemokine CCL20. The extract also significantly induced the expression of PPARγ, a ligand-activated transcription factor, and TLR2 receptor. [ABSTRACT FROM AUTHOR]

Background The domestic pig is an excellent animal model to study human microbial diseases due to its similarity to humans in terms of anatomy, physiology, and genetics. We assessed the suitability of an in vitro air-liquid interface (ALI) culture system for newborn pig trachea (NPTr) cells as a practical tool for analyzing the immune response of respiratory epithelial cells to aggressors. This cell line offers a wide microbial susceptibility spectrum to both viruses and bacteria. The purpose of our study was to evaluate and characterize diverse aspects of cell differentiation using different culture media. After the NPTr cells reached confluence, the apical medium was removed and the cells were fed by medium from the basal side. Results We assessed the cellular layer's capacity to polarize and differentiate in ALI conditions. Using immunofluorescence and electronic microscopy we evaluated the presence of goblet and ciliated cells, the epithelial junction organization, and the transepithelial electrical resistance. We found that the cellular layer develops a variable density of mucus producing cells and acquires a transepithelial resistance. We also identified increased development of cellular junctions over the culture period. Finally, we observed variable expression of transcripts associated to proteins such as keratin 8, mucins (MUC1, MUC2, and MUC4), occludin, and villin 1. Conclusions The culture of NPTr cells in ALI conditions allows a partial in vitro representation of porcine upper airway tissue that could be used to investigate some aspects of host/respiratory pathogen interactions. [ABSTRACT FROM AUTHOR]

The article offers information on a study on cardiothoracic chest ultrasound and its relevance in managing acute respiratory failure (ARF) in critical care. The results of the study shows that cardiothoracic ultrasonography is an effective complementary diagnostic tool for the evaluation of clinical, radiologic, and biologic data of ARF patients in the intensive care unit (ICU). The authors conclude that the procedure can potentially contribute to making early therapeutic decisions.

Dendritic cells (DCs) are well known as professional antigen-presenting cells (APC) able to initiate specific T-cell responses to pathogens in lymph nodes (LN) draining the site of infection. However, the respective contribution of migratory and LNresident DCs in this process remains unclear. As DC subsets represent important targets for vaccination strategies, more precise knowledge of DC subsets able to present vaccine antigens to T cells efficiently is required. To investigate the capacities of DCs migrating in the lymph (L-DCs) to initiate a specific T-cell response, we used physiologically generated DCs collected from a pseudoafferent lymphatic cannulation model in sheep. The CD1b+ L-DCs were assessed for presenting antigens from the vaccine attenuated strain of Salmonella enterica serovar Abortusovis. CD1b+ L-DCs were able to phagocytose, process and to present efficiently Salmonella antigens to effector/memory T cells in vitro. They were shown to be efficient APC for the priming of allogeneic naive T cells associated with inducing both IFN-γ and IL-4 responses. They were also efficient in presenting Salmonella antigens to autologous naive T cells associated with inducing both IFN-γ and IL- 10 responses. The capacities of L-DCs to process and present Salmonella antigens to T cells were investigated in vivo after conjunctival inoculation of Salmonella. The CD1b+ L-DCs collected after inoculation were able to induce the proliferative response of CD4+ T cells suggesting the in vivo capture of Salmonella antigens by the CD1b+ L-DCs, and their potential to present them directly to CD4+ T cells. In this study, CD1b+ L-DCs present potential characteristics of APC to initiate by themselves T cell priming in the LN. They could be used as target cells for driving immune activation in vaccinal strategies. [ABSTRACT FROM AUTHOR]

Abstract  Systems biology aims at integrating processes at various time and spatial scales into a single and coherent formal description to allow computer modeling. In this context, we focus on rule-based modeling and its integration in the domain-specific language MGS . Through the notions of topological collections and transformations, MGS allows the modeling of biological processes at various levels of description. We validate our approach through the description of various models of the genetic switch of the λ phage, from a very simple biochemical description of the process to an individual-based model on a Delaunay graph topology. This approach is a first step into providing the requirements for the emerging field of spatial systems biology which integrates spatial properties into systems biology. [ABSTRACT FROM AUTHOR]

Objective: Esophageal Doppler allows continuous monitoring of stroke volume index (SVI) and corrected flow time (FTc). We hypothesized that variations in stroke output index SOI (SVI/FTc) during volume expansion can predict the hemodynamic response to subsequent fluid loading better than the static values.Design and Setting: Prospective study in the intensive care unit of a university hospital.Patients: Fifty-one patients with circulatory failure were monitored by esophageal Doppler.Interventions: Patients who responded to a first fluid challenge received a second one. Patients who responded to both were classified as responders-responders, and those who did not respond to the second as responders-nonresponders. In these two groups we compared DeltaSVI, DeltaFTc, and DeltaSOI during each fluid challenge and also static values at the end of each fluid challenge.Measurements and Results: After the first fluid challenge DeltaSOI and DeltaSVI were significantly higher in patients who responded to subsequent volume expansion than in patients who no longer responded. ROC curves showed that DeltaSOI was a better predictor of fluid responsiveness than DeltaSVI. During volume expansion a DeltaSOI value of 11% discriminated between responders and nonresponders to subsequent volume expansion with a sensitivity of 91% and a specificity of 97%. There was no significant difference between the two groups for FTc value at the end of first fluid challenge.Conclusions: Analysis of DeltaSOI during fluid challenge predicts response to subsequent fluid challenge and FTc is not a reliable indicator of cardiac preload. [ABSTRACT FROM AUTHOR]

Provides information on a study that reported a controlled trial of treatment of hospitalized patients with severe acute alcoholic hepatitis with prednisone. Methodology of the study; Results and discussion on the study.

Dendritic cells (DCs) are pivotal in the development of specific T-cell responses to control pathogens, as they govern both the initiation and the polarization of adaptive immunity. To investigate the capacities of migrating DCs to respond to pathogens, we used physiologically generated lymph DCs (L-DCs). The flexible polarization of L-DCs was analysed in response to Salmonella or helminth secretions known to induce different T cell responses. Mature conventional CD1b+ L-DCs showed a predisposition to promote pro-inflammatory (IL-6), pro-Th1 (IL-12p40) and anti-inflammatory (IL-10) responses which were amplified by Salmonella, and limited to only IL-6 induction by helminth secretions. The other major population of L-DCs did not express the CD1b molecule and displayed phenotypic features of immaturity compared to CD1b+ L-DCs. Salmonella infection reduced the constitutive expression of TNF-α and IL-4 mRNA in CD1b- L-DCs, whereas this expression was not affected by helminth secretions. The cytokine response of T cells promoted by L-DCs was analysed in T cell subsets after co-culture with Salmonella or helminth secretion-driven CD1b+ or CD1b- L-DCs. T cells preferentially expressed the IL-17 gene, and to a lesser extent the IFN-γ and IL-10 genes, in response to Salmonella-driven CD1b+ L-DCs, whereas a preferential IL-10, IFN-γ and IL-17 gene expression was observed in response to Salmonella-driven CD1b- L-DCs. In contrast, a predominant IL-4 and IL-13 gene expression by CD4+ and CD8+ T cells was observed after stimulation of CD1b+ and CD1b- L-DCs with helminth secretions. These results show that mature conventional CD1b+ L-DCs maintain a flexible capacity to respond differently to pathogens, that the predisposition of CD1b- L-DCs to promote a Th2 response can be oriented towards other Th responses, and finally that the modulation of migrating L-DCs responses is controlled more by the pathogen encountered than the L-DC subsets. [ABSTRACT FROM AUTHOR]

A multiple internal reflection infrared monitoring system based on two-prism coupling geometry has been developed for the quantitative measurement of organic contamination on 200 or 300 mm silicon wafers. The method is rapid, simple, and can be carried out nondestructively. Varying the distance between the prisms couplers allows one to perform absolute infrared absorption measurements without the need of a reference wafer. On 300 mm silicon wafers CH[sub 2] and CH[sub 3] surface concentrations as low as 4x10[sup 12] cm[sup -2] can be measured. © 2000 American Institute of Physics. [ABSTRACT FROM AUTHOR]