Recognition and optical sensing of amines by a quartz-bound 7-chloro-4-quinolylazopillar[5]arene monolayer

Covalent bonding of 7-chloro-4-quinolylazo-octamethoxypillar[5]arene molecules to silylated quartz substrates readily produced a new chromogenic reusable pillararene-coated quartz slide, for the direct UV detection of “transparent” analytes in solution. This device provides an analyte-selective optical response towards linear (di)amines with a highly reproducible optical read-out.

Pillar[5]arene-decorated quartz slides for the direct detection of linear amines and diamines are now available.     

Next‐generation sequencing for the diagnosis of MYH9‐RD: Predicting pathogenic variants

The heterogeneous manifestations of MYH9‐related disorder (MYH9‐RD), characterized by macrothrombocytopenia, Döhle‐like inclusion bodies in leukocytes, bleeding of variable severity with, in some cases, ear, eye, kidney, and liver involvement, make the diagnosis for these patients still challenging in clinical practice. We collected phenotypic data and analyzed the genetic variants in more than 3,000 patients with a bleeding or platelet disorder. Patients were enrolled in the BRIDGE‐BPD and ThromboGenomics Projects and their samples processed by high throughput sequencing (HTS). We identified 50 patients with a rare variant in MYH9. All patients had macrothrombocytes and all except two had thrombocytopenia. Some degree of bleeding diathesis was reported in 41 of the 50 patients. Eleven patients presented hearing impairment, three renal failure and two elevated liver enzymes. Among the 28 rare variants identified in MYH9, 12 were novel. HTS was instrumental in diagnosing 23 patients (46%). Our results confirm the clinical heterogeneity of MYH9‐RD and show that, in the presence of an unclassified platelet disorder with macrothrombocytes, MYH9‐RD should always be considered. A HTS‐based strategy is a reliable method to reach a conclusive diagnosis of MYH9‐RD in clinical practice.     

Occupational asthma: role of airway inflammation and remodelling in persistent respiratory symptoms and bronchial hyper-responsiveness

BACKGROUND: Until the late 1970's occupational asthma (OA) was considered reversible once patients were removed from exposure. Unfortunately, respiratory symptoms and non-specific bronchial hyper-responsiveness (NSBH) persist in about two-thirds of patients for years after removal from the offending agent. OBJECTIVES AND METHODS: This review focuses on the role of airways inflammation and remodelling in persistent respiratory symptoms and NSBH after cessation of occupational exposure. RESULTS: Even though cessation of exposure does not always result in remission of OA, symptoms, airways calibre and NSBH do improve in many patients. Although improvements in FEV1 and NSBH tend to reach a plateau 1-2 years after workers leave exposure, reversing NSBH may take much longer and respiratory symptoms and NSBH can persist in subjects removed from exposurefor >10 yrs. Long-term treatment with inhaled corticosteroids (ICS) induces a small but significant improvement in respiratory symptoms and in quality of life and a decrease in NSBH. Prolonged exposure and respiratory symptoms, marked airway obstruction and NSBH, high total cell, eosinophil and neutrophil counts in bronchoalveolar lavage fluid, a strong reaction during specific inhalation challenge, and delayed treatment with ICS have been identified as prognostic factors of unfavourable outcome. If exposure persists, OA tends to deteriorate in many patients but regular long-term treatment with ICS and long-acting beta2-agonists seems to stabilize the outcome. Soon after the last exposure inflammatory cell infiltrates, including eosinophils, and increased thickness of sub-epithelial collagen have been observed. When time since removal from exposure was longer, persistence of respiratory symptoms and NSBH was associated with airway inflammation, remodelling and hypersensitivity to the offending agent. Thickness of sub-epithelial collagen and specific airway sensitivity were reduced after prolonged non-exposure to isocyanates, although NSBH and airway inflammation persisted. CONCLUSIONS: Pathologic features are similar in OA and non-occupational asthma. The main factors of favourable outcome are early removal from exposure and a mild airway obstruction and NSBH at diagnosis. Persistence of airway inflammation years after removal from exposure suggests this process may become independent of the offending agent. The role of remodelling on persistence of OA needs to be clarified further.     

Prognostic value of thyrotropin receptor antibodies (TRAb) in Graves' disease: a 120 months prospective study

In most trials, at least 30-60% of patients with Graves' disease treated with antithyroid drugs relapse within 2 years after therapy withdrawal. At present, there are no prognostic parameters available early in treatment to indicate patients likely to achieve long-term remission. Because thyrotropin receptor autoantibodies (TRAb) are specific for Graves' disease, we evaluated the ability of their levels and of their rate of change to predict long-term prognosis. In our study 216 consecutive patients with newly diagnosed Graves' disease started a therapy with methimazole. Patients were treated until they achieved euthyroidism and TRAb were measured at 6-month intervals throughout a follow up of 120 months. Our study demonstrated that at the onset of hyperthyroidism patients' age, sex, fT4 levels and goiter size had no prognostic value in predicting long-term prognosis (respectively p = 0.79; p = 0.98; p = 0.83; p = 0.89). On the contrary, at the time of diagnosis TRAb titer was a good predictor of the final outcome (p<0.001); a titer equal to (or) more than 46.5 UI/L could identify patients who had never achieved long-term remission with a sensitivity of 52% and a specificity of 78%. Also fall rate of TRAb at 6 months of follow up and after therapy withdrawal were useful to predict the final outcome (p<0.001). At 6 months of follow up the time of therapy withdrawal, a decrease of TRAb lower than 52.3% or even its increase could identify patients who had never achieved permanent remission with a sensitivity of 55% and a specificity of 79.1%. No single parameter among TRAb, satisfactory identified a sub-set of patients who achieved long remission. Accordingly to our data, the best result in predicting long term remission is probably given by the presence of at least one of the two features evaluated at 6 months (TRAb titer and/or percentage of TRAb fall rate), with a sensitivity of 63% and specificity of 88%. TRAb titers evaluated both at the onset of hyperthyroidism that at 6 months of therapy or their rate of fall at 6 months and at ATD withdrawal are predictors of outcome. However, the presence of at least one, between titers of TRAb or their rate of fall at six months, resulted to be the best predictor of remission with the higher sensitivity and specificity.     

Expression and role of CCR6/CCL20 chemokine axis in pulmonary sarcoidosis

We have shown previously that the chemokine receptors CXCR3 and CXCR6 are coexpressed by Th1 cells infiltrating the lung and the granuloma of patients with sarcoidosis. In this study, we evaluated the role of CCL20/CCR6 interaction in the pathogenesis of acute and chronic pulmonary sarcoidosis. By flow cytometry and molecular analyses, we have demonstrated that Th1 cells isolated from the bronchoalveolar lavage (BAL) of patients with sarcoidosis and T cell alveolitis are equipped with CCR6. Furthermore, CCR6(+) T cells coexpressed the chemokine receptors CXCR3 and CXCR6. Immunohistochemical analysis of lung specimens has shown that CCR6(+) T cells infiltrate lung interstitium and surround the central core of the granuloma. It is interesting that CCR6 was never detected on the alveolar macrophage (AM) surface, and it is observed in the cytoplasm of AMs from patients with sarcoidosis and alveolitis. The CCR6 ligand CCL20 was expressed by macrophages, multinucleated giant cells, and epithelioid cells infiltrating the granuloma. Furthermore, detectable levels of CCL20 protein are seen in the BAL fluid components of patients with active sarcoidosis, and sarcoid AMs release the CCR6 ligand in vitro. From a functional point of view, sarcoid Th1 cells were able to respond to CXCL10, CXCL16, and CCL20 in migratory assays. In vitro kinetic studies demonstrated that CCR6 is induced rapidly by IL-2, IL-18, and IFN-gamma. In conclusion, T cells expressing CCR6, CXCR3, and CXCR6 act coordinately with respective ligands and Th1 inflammatory cytokines in the alveolitic/granuloma phases of the disease.