contribution of bronchial biopsies in the evaluation of pathogenesis and progression of copd

This review summarizes and discusses the lung pathology of COPD patients emphasising on inflammatory cell phenotypes and mechanisms which prevail in different clinical conditions. In bronchial biopsies a series of events takes place during the progression of the disease from mild to severe. T-lymphocytes, particularly CD8+ cells and macrophages are the prevalent inflammatory cells in the lungs of healthy smokers and patients with mild/moderate COPD. This T-cell activation seems to be sustained by CD4+, CD8+ cells and macrophages expressing transcription factors and Tc1 cytokines such as NF-kB, STAT4 and IFNγ. In contrast, severe disease is characterized by lymphocytes producing greater amounts of TGF-ß1 and by an increase of nitrotyrosine immunoreactivity and activated neutrophils, macrophages and MPO+ cells. However, the mechanisms involved in neutrophilic migration and adhesion are currently under investigation. Recent data has shown that in severe COPD there is an impaired neutrophil capability to respond to chemotactic stimuli, as well as an increased collagen adhesion of neutrophils due to the up-regulation of CD44 and CD11b receptors. This data together, may account for the increased neutrophilia observed in the severe disease states of COPD. In this context, insights obtained from the tissutal analysis of bronchial biopsies represent an irreplaceable route to further progresses in to the pathogenesis of this disease

Brown Tumour in a Patient with Secondary Hyperparathyroidism Resistant to Medical Therapy: Case Report on Successful Treatment after Subtotal Parathyroidectomy

Brown tumour represents a serious complication of hyperparathyroidism. Differential diagnosis, based on histological examination, is only presumptive and clinical, radiological and laboratory data are necessary for definitive diagnosis. Here we describe a case of a brown tumour localised in the maxilla due to secondary hyperparathyroidism in a young women with chronic renal failure. Hemodialysis and pharmacological treatment were unsuccessful in controlling secondary hyperparathyroidism making it necessary to proceed with a subtotal parathyroidectomy. The proper timing of the parathyroidectomy and its favourable effect on regression of the brown tumor made it possible to avoid a potentially disfiguring surgical removal of the brown tumor

The role of autophagy in vernal kerato-conjunctivitis

Autophagy is involved in many biological aspects, including cell survival and death, innate and adaptive immunity and cancer. An involvement of autophagy is also reported in some inflammatory diseases such as asthma. In the present study we explored the role of autophagy in vernal keratoconjunctivitis (VKC), a severe inflammatory disease mainly found in children and adolescents. Autophagy and apoptosis markers (LC3A, LC3B, Beclin-1, cathepsin B, BCL-2, BAX, caspase 3) expression in conjunctival biopsies from 9 active VKC patients and 9 healthy age matched normal subjects were analyzed using immunohistochemistry and qPCR techniques. Conjunctival cells cultures were treated with inflammatory stimuli (IL-1b, histamine, IL-4, TNFa) and analysed by western blotting for autophagy markers expression. LC3B, Catepsin D and B and Beclin-1expression strongly increased in the stroma of VKC whereas the epithelium was consistently negative for all of the molecules studied but positive for Beclin-1 in VKC. qPCR analysis demonstrated a similar mRNAs expression in VKC and normal subjects. In “in vitro” experiments autophagy induction revealed that only LC3B expression was changed in conjunctival fibroblasts by inflammatory stimuli. In particular, both LC3BI, the LC3B free form, and LC3BII, the phosphatidyl-ethanolamine-conjugated form, involved in the autophagosome formation, were decreased in fibroblast cultures at 24h after TNFα stimulation. However, since LC3B-II is normally degraded by lysosomes and the total amount of LC3B-II depends on the balance between its formation and degradation, we analyzed the expression of LC3B-II in the presence and absence of chloroquine, an inhibitor of lysosomal degradation. We found a significant increased amount of LC3BII compared to the control, indicating an over-expression of this protein in stimulated fibroblasts that is quickly damped by its degradation. Since one of the key steps in autophagy is the conversion of LC3B from LC3B-I to LC3B-II, our results suggest that autophagy may be involved in the pathogenesis of VKC

Possible Autophagy induction in Vernal Keratoconjunctivitis via Tumor Necrosis Factor Alpha Stimulation

Tumor necrosis factor alpha (TNFα) is one of the main mediators of inflammatory response in many pathological diseases, involved in a widespread biological functions, including autophagy. Previous data obtained in our laboratory demonstrated that TNFα and some autophagy markers (which markers please indicate) are overexpressed in a severe inflammatory disease such as vernal keratoconjunctivitis (VKC). In the present study we explored the role of TNFα in the induction of autophagy in VKC, using an in vitro model. Primary conjunctival cell cultures were treated with TNFa and analysed by qPCR and western blotting for expression of some autophagy and lysosomial markers at 4, 10 and 24 hours after exposure. qPCR results demonstrated that LC3B, Beclin-1, LAMP1 and p62 strongly increased from 4 to 24 hours, whereas the expression of Catepsin D, a protein implicated in lysosomial apototic pathway, was comparable to that of untreated control. Western blotting analysis revealed lipidation of LC3B quantified as an increased LC3BII/LC3BI ratio. Moreover, double immunofluorescence for Cathepsin D and LAMP1 showed that Cathepsin D was localized within the lysosomes at 4, 10, 24 hours after cell exposure to inflammatory stimuli. In conclusion, our data demonstrated that TNFα significantly induce in VKC LC3B lipidation, LC3BII/LC3BI ratio and p62 (qPCR) in the cells exposed to inflammatory stimuli which shows possible activation of autophagy pathway

4D Printing of Plasmon-Encoded Tunable Polydimethylsiloxane Lenses for On-Field Microscopy of Microbes

Here the 4D printing of a magnifying polydimethylsiloxane (PDMS) lens encoded with a tunable plasmonic rejection filter is reported. The lens is formed by moldless printing of PDMS pre-polymer on a nanostructured porous silicon (PSi) templating layer. A nanometer-thick plasmonic filter is integrated on the lens surface by in situ synthesis of Ag and Au nanoparticles (NPs) with programmed density. The filter can be designed to reject light at the plasmonic resonance wavelength of the NPs with an optical density tunable from 0 to 3 and retreive light at longer wavelengths with a pass-to-stop band ratio tunable from 0 to 60 dB. Swelling of PDMS in hexane and ether is used to change the NP density on the lens surface and modulate, in turn, the transmittance properties of the NP-decorated lens over 3 orders of magnitude. The plasmon-encoded lens is coupled to a commercial smartphone demonstrating: shaping of the emission spectrum of a white light-emitting diode to tune the color from yellow to purple; real-time bright-field and fluorescence microscopy of living microbes in water, namely, the auto-fluorescent green alga Chlorogonium sp. and the ciliated protozoan Euplotes daidaleos

Integration of HVSR measures and stratigraphic constraints for seismic microzonation studies: the case of Oliveri (ME)

Because of its high seismic hazard the urban area of Oliveri has been subject of first level seismic microzonation. The town develops on a large coastal plain made of mixed fluvial/marine sediments, overlapping a complexly deformed substrate. In order to identify points on the area probably suffering relevant site effects and define a preliminary Vs subsurface model for the first level of microzonation, we performed 23 HVSR measurements. A clustering technique of continuous signals has been used to optimize the calculation of the HVSR curves. 42 reliable peaks of the H/V spectra in the frequency range 0.6–10 Hz have been identified. A second clustering technique has been applied to the set of 42 vectors, containing Cartesian coordinates, central frequency and amplitude of each peak to identify subsets which can be attributed to continuous spatial phenomena. The algorithm has identified three main clusters that cover significant parts of the territory of Oliveri. The HVSR data inversion has been constrained by stratigraphic data of a borehole. To map the trend of the roof of the seismic bedrock, from the complete set of model parameters only the depth of the seismic interface that generates peaks fitting those belonging to two clusters characterized by lower frequency has been extracted

The role of heat shock proteins in the inflammatory state of vernal keratoconjunctivitis

The aim of the study was to analyse the role of heat shock proteins (HSPs) in vernal keratoconjunctivitis (VKC), a recurrent allergic ocular inflammatory disease. We evaluated the expression of some HSPs (Hsp10, Hsp27, Hsp40, Hsp60, Hsp70, Hsp90) in the mucosal biopsies of VKC patients by immunohistochemistry, and in conjunctival cells cultures treated with inflammatory stimuli (IL-1β, histamine, IL-4, TNFβ, UVB irradiation) by western blotting. Immunohistochemical analysis revealed that Hsp10, Hsp27, Hsp40, Hsp70 and Hsp90 expression was significantly increased in VKC whereas the Hsp60 level was unaltered. In vitro induction by inflammatory stimuli in Chang epithelial conjunctival cells revealed that Hsp70 protein expression was significantly increased in epithelial cells line after 4-10 h from histamine and IL-4 stimulation. The same molecule was also overexpressed in conjunctival fibroblast cultures after TNFβ treatment. Hsp90 protein level was increased in the same cell cultures by IL-1β at 4-10-24 h. The Hsp40 protein expression was increased both in epithelial and fibroblast cultures induced by all inflammatory stimuli. Moreover, UVB irradiation significantly increased Hsp90 expression in primary fibroblast culture and Hsp27 in conjunctival epithelial cells after 10 hours. These results indicate that HSPs levels increase in VKC. In particular, Hsp40 expression is up-regulated by all the typical inflammatory stimuli involved in VKC pathogenesis. The specific role of each one of these chaperonins to further induce or counteract inflammation need to be further investigated