Сжигание или мелкое диспергирование створок головного обтекателя ракеты-носителя за счет дополнительного подвода тепла при движении на атмосферном участке траектории спуска

The aim of this study was the establishment of precision-cut lung slices (PCLS) as a suitable ex vivo alternative approach to animal experiments for investigation of immunomodulatory effects. For this purpose we characterized the changes of cytokine production and the expression of cell surface markers after incubation of PCLS with immunoactive substances lipopolysaccharide (LPS), macrophage-activating lipopeptide-2 (MALP-2), interferon ? (IFN?), and dexamethasone. Viability of PCLS from wild-type and CD11c-enhanced yellow fluorescent protein (CD11-EYFP)-transgenic mice was controlled by measurement of lactate dehydrogenase (LDH) enzyme activity and live/dead fluorescence staining using confocal microscopy. Cytokines and chemokines were detected with Luminex technology and ELISA. Antigen presenting cell (APC) markers were investigated in living mouse PCLS in situ using confocal microscopy. LPS triggered profound pro-inflammatory effects in PCLS. Dexamethasone prevented LPS-induced production of cytokines/chemokines such as interleukin (IL)-5, IL-1greek small letter alpha, TNFgreek small letter alpha, IL-12(p40), and RANTES in PCLS. Surface expression of MHC class II, CD40, and CD11c, but not CD86 was present in APCs of naive PCLS. Incubation with LPS enhanced specifically the expression of MHC class II on diverse cells. MALP-2 only failed to alter cytokine or chemokine levels, but was highly effective in combination with IFN? resulting in increased levels of TNFgreek small letter alpha, IL-12(p40), RANTES, and IL-1greek small letter alpha. PCLS showed characteristic responses to typical pro-inflammatory stimuli and may thus provide a suitable ex vivo technique to predict the immunomodulatory potency of inhaled substances

Импортозамещение межсекционных уплотнений на примере многоступенчатого насоса "Grundfos"

Bronchoconstriction is a characteristic symptom of various chronic obstructive respiratory diseases such as chronic obstructive pulmonary disease (COPD) and asthma. Precision-cut lung slices (PCLS) are a suitable ex vivo model to study physiological mechanisms of bronchoconstriction in different species. In the present study, we established an ex vivo model of bronchoconstriction in non-human primates (NHPs). PCLS prepared from common marmosets, cynomolgus macaques, rhesus macaques, and anubis baboons were stimulated with increasing concentrations of representative bronchoconstrictors: methacholine, histamine, serotonin, leukotriene D4 (LTD4), U46619, and endothelin-1. Alterations in the airway caliber were measured and compared to previously published data from rodents, guinea pigs, and humans. Methacholine induced maximal airway constriction, varying between 74 and 88% in all NHP species, whereas serotonin was ineffective. Histamine induced maximal bronchoconstriction of 77 to 90% in rhesus macaques, cynomolgus macaques, and baboons, and a lesser constriction of 53% in marmosets. LTD4 was ineffective in marmosets and rhesus macaques, but induced a maximum constriction of 44 to 49% in cynomolgus macaques and baboons. U46619 and endothelin-1 caused airway constriction in all NHP species, with maximum constrictions of 65 to 91%, and 70 to 81%, respectively. In conclusion, PCLS from NHPs represent a valuable ex vivo model for studying bronchoconstriction. All NHPs respond to mediators relevant to human airway disorders such as methacholine, histamine, U46619, endothelin-1 and are insensitive to the rodent mast cell product serotonin. Only PCLS from cynomolgus macaques and baboons, however, responded also to leukotrienes, suggesting that among all compared species, these two NHPs resemble the human airway mechanisms bes

Effects of the TLR2 Agonists MALP-2 and Pam3Cys in Isolated Mouse Lungs

Background: Gram-positive and Gram-negative bacteria are main causes of pneumonia or acute lung injury. They are recognized by the innate immune system via toll-like receptor-2 (TLR2) or TLR4, respectively. Among all organs, the lungs have the highest expression of TLR2 receptors, but little is known about the pulmonary consequences of their activation. Here we studied the effects of the TLR2/6 agonist MALP-2, the TLR2/1 agonist Pam 3Cys and the TLR4 agonist lipopolysaccharide (LPS) on pro-inflammatory responses in isolated lungs. Methodology/Principal Findings: Isolated perfused mouse lungs were perfused for 60 min or 180 min with MALP-2 (25 ng/ mL), Pam3Cys (160 ng/mL) or LPS (1 mg/mL). We studied mediator release by enzyme linked immunosorbent assay (ELISA), the activation of mitogen activated protein kinase (MAPK) and AKT/protein kinase B by immunoblotting, and gene induction by quantitative polymerase chain reaction. All agonists activated the MAPK ERK1/2 and p38, but neither JNK or AKT kinase. The TLR ligands upregulated the inflammation related genes Tnf, Il1b, Il6, Il10, Il12, Ifng, Cxcl2 (MIP-2a) and Ptgs2. MALP-2 was more potent than Pam 3Cys in inducing Slpi, Cxcl10 (IP10) and Parg. Remarkable was the strong induction of Tnc by MALP2, which was not seen with Pam 3Cys or LPS. The growth factor related genes Areg and Hbegf were not affected. In addition, all three TLR agonists stimulated the release of IL-6, TNF, CXCL2 and CXCL10 protein from the lungs

Precision cut lung slices as tool for assessment of allergens

Studies on precision cut lung slices (PCLS) are part of the European Union project SENS-IT-IV. Aim of this project is the development and establishment of in vitro techniques for assessment of allergenic potential of inhaled substances without animal testing. Mouse PCLS were prepared and treated with subtoxic concentrations of respiratory (ammonium-hexachloroplatinate, AHCP) and contact (cinnamaldehyde) allergens in comparison to negative control (salicylic acid). Supernatants were collected, PCLS were lysed and extrinsic and intrinsic cytokine profiles were analysed by ELISA and cytokine array. In addition lung slices were stained with calcein and anti-ICAM-1 for confocal microscopy. Cytokine levels showed significant increases after treatment with AHCP compared to tissue control. Interleukine (IL)-10 was increased up to 2.2-fold, TNF-alpha and IL-1alpha up to 1.6-fold and MIP-1beta up to 0.7-fold. Cytokines like eotaxin, G-CSF and IL-12 showed a 0.75-fold increase. Stimulation with cinnamaldehyde resulted in a significant decrease in MIP-1beta up to 0.5-fold, but showed no significant changes in expression of other cytokines. Salicylic acid showed compared to AHCP an increase of TNF-alpha-expression, but e.g. no increase of IL-1alpha. Staining of epithelial cells showed an upregulation of ICAM-1 in response to activation. Our experiments showed different expression patterns of cytokines for representative respiratory and contact allergens and indicate that PCLS is a usable ex vivo model for the assessment of new substances

Respiratory toxicology and immunotoxicology in human precision cut lung slices (PCLS)

Occupational asthma is one of the most common lung diseases in developed countries. Risk assessment for potentially sensitising chemicals is performed in animal models. With an increasing public demand to limit the number of animals used in respiratory research and to reduce the distress to the animals, several models have been developed. Human PCLS are an ex vivo model where all relevant cell types are present in their natural position. We use PCLS to test for modifications of local immune responses assessing a variety of immunological endpoints. Human PCLS were prepared from lung lobes of cancer patients. Tissue was exposed to LPS, dexamethasone, respiratory and contact allergens. Viability of PCLS was determined with WST-1, LDH and LIVE/DEAD staining for confocal microscopy. Cytokine contents were detected with Luminex technology and ELISA. Employing LPS and dexamethasone we were able to show that the inflammatory response in PCLS resemble the in vivo situation very closely. Repeated stimulation with LPS (intra- and inter-assay variances <20%) showed that human PCLS might also be suitable to characterize respiratory irritation and inflammation induced by chemicals. PCLS were exposed to 20 chemicals and IC50 were calculated. We currently investigate cytokine patterns (e.g. IL-1, TNF, IL-8) for the differentiation between respiratory and contact allergens. Indeed, IL-8 production is increased after stimulation with TMA whereas DNCB failed to induce the release of IL-8 to the same extent. It suggests that the combination of cytokine production with cytotoxic data may represent a promising in vitro model for the screening of allergens