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

An Optimized Procedure for the Site-Directed Labeling of NGF and proNGF for Imaging Purposes

Neurotrophins are growth factors of fundamental importance for the development, survival and maintenance of different neuronal and non-neuronal populations. Over the years, the use of labeled neurotrophins has helped in the study of their biological functions, leading to a better understanding of the processes that regulate their transport, traffic, and signaling. However, the diverse and heterogeneous neurotrophin labeling strategies adopted so far have often led to poorly reproducible protocols and sometimes conflicting conclusions. Here we present a robust, reliable, and fast method to obtain homogeneous preparations of fluorescent proNGF and NGF with 1:1 labeling stoichiometry. This strategy is well suited for several applications, ranging from advanced imaging techniques such as single particle tracking, to analyses that require large amounts of neurotrophins such as in vivo monitoring of protein biodistribution. As a proof of the quality of the labeled NGF and proNGF preparations, we provide a quantitative analysis of their colocalization with proteins involved in the signaling endosome function and sorting. This new analysis allowed demonstrating that proNGF localizes at a sub-population of endosomes not completely overlapped to the one hosting NGF

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

Influence of Myeloperoxidase Levels on Periodontal Disease: An Applied Clinical Study

In this trial, we evaluated the influence on plasma and salivary myeloperoxidase (MPO) levels of periodontal health, coronary heart disease (CHD), periodontitis, or both periodontitis and CHD. Clinical and periodontal parameters were collected from periodontitis patients (n = 31), CHD patients (n = 31), patients with both periodontitis and CHD (n = 31), and from healthy patients (n = 31) together with saliva and plasma samples. The median concentrations of salivary and plasma MPO were statistically higher in the CHD patients [plasma: 26.2 (18.2–34.4) ng/mg; saliva 83.2 (77.4–101.5) ng/mL, p < 0.01] and in the periodontitis plus CHD patients [plasma: 27.8 (22.5–35.7) ng/mg; saliva 85.6 (76.5–106.7) ng/mL, p < 0.001] with respect to periodontitis and control patients. Through a univariate regression analysis, c-reactive protein (CRP) and CHD (both p < 0.001) and periodontitis (p = 0.024) were statistically correlated with MPO in plasma. The multivariate regression analysis demonstrated that only CRP was statistically the predictor of MPO in plasma (p < 0.001). The multivariate regression analysis in saliva demonstrated that, regarding MPO levels the only predictors were CRP (p < 0.001) and total cholesterol (p = 0.035). The present study evidenced that subjects with CHD and periodontitis plus CHD had higher plasma and salivary levels of MPO compared to subjects with periodontitis and healthy controls

Efgartigimod beyond myasthenia gravis: the role of FcRn-targeting therapies in stiff-person syndrome

Stiff-person syndrome (SPS) is a rare autoimmune neurological disorder characterized by high titers of antibodies against glutamic acid decarboxylase (GAD) causing impaired GABAergic inhibitory neurotransmission. To date, there is not a defined therapy for such condition, but immunomodulating therapies, such as plasma exchange, intravenous immunoglobulins, and rituximab, have been widely used in clinical practice. However, the efficacy and tolerability of these treatments is not well established. Efgartigimod, a new neonatal Fc receptor (FcRn) blocker, is a human IgG1 antibody Fc fragment engineered with increased affinity for FcRn binding, leading to a reduction in IgGs levels, including pathogenic IgG autoantibody showing promising results in neurological autoimmune disorders and has been approved for the treatment of AChR-seropositive generalized myasthenia gravis (MG). In this study, we report and describe the first data on treatment with efgartigimod in three patients affected by both AChR-seropositive generalized MG and anti-GAD-seropositive SPS. Patients were followed since the start of efgartigimod and for the whole treatment period (12 weeks). MG symptoms were assessed with the "MG activity of daily living score" and the Quantitative Myasthenia Gravis score, while SPS ones were assessed with the "SPS activity of daily living score"; muscle strength was assessed with the Medical Research Council Sum score; the overall disability from MG and SPS was assessed by the modified Rankin Scale. All patients showed an improvement in symptoms of both SPS and MG after 2 cycles of treatment. Our data suggest that efgartigimod may be considered as a candidate drug for SPS and other autoantibody-mediated neurological disorders

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

Effects of artificial canopy shading on vegetative growth and ripening processes of cv. Nero d’Avola (Vitis vinifera L.)

Introduction: The biology of the grapevine (Vitis vinifera L.) is clearly influenced by the climatic conditions of the growing environment, where temperature and light play a major role in modifying plant physiology. In the scenario of climatic changes, radiative excess, correlated to the increase in temperature, can concretely subject the photosynthetic apparatus to a condition of light saturation and cause a drastic reduction in photochemical efficiency, giving rise to chronic photoinhibition phenomena. Undoubtedly, the ripening behavior also undergo evident alterations; the problem of sugar ripening, which is often strongly accelerated, is induced not only by high temperatures but also by the excess concentration of carbon dioxide (CO2), which results in a higher ripening. In addition, high berry temperatures favor a reduction in the concentration of organic acids. The reported trends indicate that the need for urgent action is closely linked to the future environmental impacts that these changes could have on the entire wine sector. In recent years, shade treatments have been applied to the vine canopy to overcome this issue. Methods: The objective of this study was to determine how artificial canopy shading affects the vines vegetative growth and the ripening processes of Vitis vinifera cv. Nero d’Avola during the 2019-2020 vegetative seasons. Three treatments were established: shading treatment with a green net (shade factor 27%), shading treatment with a white net (shade factor 32%), and untreated vines, thus naturally exposed to light radiation. Results and discussion: Artificial shading, applied at full fruit set, interfered with the microclimate of the vines, causing partial effects on the grape ripening processes. At harvest, significant differences were found between the treatments in terms of sugars, also shading treatments increased must acidity and decrease pH. Results obtained on vegetative parameters, suggest that the shading treatment delays leaf fall, with potentially positive effects on the starch accumulation on perennial reserve organs to be exploited at the following season’s sprouting. Shading significantly reduced berry size, with obvious consequences on bunch weight and yield per vine. In 2020, shaded plants showed a delay in all the phenological phases. The total anthocyanins content was not changed by the shading treatment. The results obtained confirm the importance of net coverage on the microclimate of the vines, vegetative-productive activity, and grapes quality. From this point of view, the net covering technique can be a tool for controlling grapes ripening dynamics in the context of climate change

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