Virtual Screening Strategy and In Vitro Tests to Identify New Inhibitors of the Immunoproteasome

Immunoproteasome inhibition is a promising strategy for the treatment of hematological malignancies, autoimmune diseases, and inflammatory diseases. The design of non-covalent inhibitors of the immunoproteasome beta 1i/beta 5i catalytic subunits could be a novel approach to avoid the drawbacks of the known covalent inhibitors, such as toxicity due to off-target binding. In this work, we report the biological evaluation of thirty-four compounds selected from a commercially available collection. These hit compounds are the outcomes of a virtual screening strategy including a dynamic pharmacophore modeling approach onto the beta 1i subunit and a pharmacophore/docking approach onto the beta 5i subunit. The computational studies were first followed by in vitro enzymatic assays at 100 mu M. Only compounds capable of inhibiting the enzymatic activity by more than 50% were characterized in detail using Tian continuous assays, determining the dissociation constant (K-i) of the non-covalent complex where K-i is also the measure of the binding affinity. Seven out of thirty-four hits showed to inhibit beta 1i and/or beta 5i subunit. Compound 3 is the most active on the beta 1i subunit with K-i = 11.84 +/- 1.63 mu M, and compound 17 showed K-i = 12.50 +/- 0.77 mu M on the beta 5i subunit. Compound 2 showed inhibitory activity on both subunits (K-i = 12.53 +/- 0.18 and K-i = 31.95 +/- 0.81 on the beta 1i subunit and beta 5i subunit, respectively). The induced fit docking analysis revealed interactions with Thr1 and Phe31 of beta 1i subunit and that represent new key residues as reported in our previous work. Onto beta 5i subunit, it interacts with the key residues Thr1, Thr21, and Tyr169. This last hit compound identified represents an interesting starting point for further optimization of beta 1i/beta 5i dual inhibitors of the immunoproteasome

Clinical features and outcomes of elderly hospitalised patients with chronic obstructive pulmonary disease, heart failure or both

Background and objective: Chronic obstructive pulmonary disease (COPD) and heart failure (HF) mutually increase the risk of being present in the same patient, especially if older. Whether or not this coexistence may be associated with a worse prognosis is debated. Therefore, employing data derived from the REPOSI register, we evaluated the clinical features and outcomes in a population of elderly patients admitted to internal medicine wards and having COPD, HF or COPD + HF. Methods: We measured socio-demographic and anthropometric characteristics, severity and prevalence of comorbidities, clinical and laboratory features during hospitalization, mood disorders, functional independence, drug prescriptions and discharge destination. The primary study outcome was the risk of death. Results: We considered 2,343 elderly hospitalized patients (median age 81 years), of whom 1,154 (49%) had COPD, 813 (35%) HF, and 376 (16%) COPD + HF. Patients with COPD + HF had different characteristics than those with COPD or HF, such as a higher prevalence of previous hospitalizations, comorbidities (especially chronic kidney disease), higher respiratory rate at admission and number of prescribed drugs. Patients with COPD + HF (hazard ratio HR 1.74, 95% confidence intervals CI 1.16-2.61) and patients with dementia (HR 1.75, 95% CI 1.06-2.90) had a higher risk of death at one year. The Kaplan-Meier curves showed a higher mortality risk in the group of patients with COPD + HF for all causes (p = 0.010), respiratory causes (p = 0.006), cardiovascular causes (p = 0.046) and respiratory plus cardiovascular causes (p = 0.009). Conclusion: In this real-life cohort of hospitalized elderly patients, the coexistence of COPD and HF significantly worsened prognosis at one year. This finding may help to better define the care needs of this population

Poteri ordinari e straordinari nella gestione dei pericoli ambientali

The judgment under review provides a valuable opportunity to reflect on the tools available to public administrations to cope with situations of environmental danger and in particular on the mayor’s exercise of the power of extra ordinem ordinance, as provided for in Articles 50 and 54 D.Lgs. n. 267/2000, following reports from private individuals regarding the aforementioned dangers

Early diagnosis of vertebral fractures

Vertebral fractures are a common clinical entity, caused by trauma or related to osteoporosis (benign). Their recognition is especially important in the post-menopausal female population but also important is their differentiation from pathological (malignant) fractures (1). A vertebral fracture is evidenced by vertebral body deformity or reduction in vertebral body height beyond a certain threshold value in the absence of bone discontinuity. For prognosis and treatment it is extremely important to recognize the cause of the fracture. In contrast to fractures that occur in other locations, vertebral fractures often go unrecognized in the acute phase as the pain may be transient and radiographic and evaluation of the spine may be difficult (2). Objective measurement of the vertebral deformity provides invaluable information to the interpreting physician and helps grade fracture severity. The recognition and diagnosis of vertebral fractures can be performed using additional diagnostic tool

Non-covalent immunoproteasome inhibitors: virtual screening and in vitro test on β1i /β5i subunits

Immunoproteasome inhibition is a challenging strategy for the treatment of hematological malignancies, autoimmune and inflammatory diseases [1,2]. The search for non-covalent inhibitors of the immunoproteasome β1i/β5i catalytic subunits could be a new strategy to avoid the drawbacks of the known covalent inhibitors. Here, we report the biological evaluation of thirty-four compounds selected from commercial libraries. A virtual screening strategy including a dynamic pharmacophore modeling approach onto the β1i subunit and a pharmacophore/docking approach onto the β5i subunit aided the identification of these hits [3]. Compound 3 is the most active onto β1i subunit with Ki = 11.84±1.63 μM, compound 17 showed Ki = 12.50±0.77 μM onto β5i subunit. Compound 2 showed inhibitory activity on both subunits (Ki = 12.53±0.18 Ki = 31.95±0.81 onto β1i subunit and β5i subunit, respectively). The hit compounds identified represent an interesting starting point for further optimization

Occult breast cancer in a female with benign lesions

Occult breast cancer is a carcinoma discovered by the presence of axillary lymph node metastases without the detection of the primary breast tumor. The incidence of this very rare pathology is 0.3%-1.0%. The limited number of these cases does not allow for the precise management of this rare pathology and often, the breast cancer manifestation can take many years to become obvious. We report the case of a 35-year-old woman who presented to our department for annual breast screening examination, without any symptoms. At the time of visit, there were two right and one left tumefactions; unfixed and palpable. Ultrasonography examination confirmed the lesions to be benign. One year later, a palpable hypoechoic axillary left lesion appeared: a lymph node with doubtful morphology. On cytological examination, a biopsy was performed for the axillary left mass which showed irregular masses of large malignant cells with pleomorphism and mitotic figures that suggested a carcinoma. The management of this case is suggestive for cancer of unknown primary syndrome