N-FORMYL PEPTIDE RECEPTORS AND UPAR FOSTER THE OXIDATIVE STRESS IN SYSTEMIC SCLEROSIS

Systemic sclerosis (SSc) is characterized by chronic inflammation and fibrosis. N-Formyl peptide receptors (FPRs) are chemotactic receptors involved in inflammation. Three FPRs have been identified: FPR1, FPR2, and FPR3. We have examined, by RT-PCR, Western blot and immunohistochemistry, FPRs expression in skin fibroblasts from normal subjects and SSc patients, showing an increased expression in SSc fibroblasts. Several functions of FPRs occur through the interaction with the chemotactic domain (residues 88SRSRY92) of the urokinase-type plasminogen activator receptor (uPAR). Western blot and immunohistochemistry analysis also showed an increased expression in SSc fibroblasts of a cleaved uPAR form exposing the SRSRY sequence at its N-terminus (DIIDIII-uPAR88–92). We demonstrated that FPRs stimulation promoted radical oxygen species (ROS) generation in normal and SSc fibroblasts. Upon stimulation, ROS production was due to FPRs interaction with uPAR and to beta 1 integrin engagement. FPRs cross-talk with uPAR and integrins led to Rac1 and ERKs activation. Rac1 is a cytosolic component of the NADPH oxidase system. Active GTP-Rac1 binds to p67phox that translocates to the membrane and associates with the catalytic gp91phox. FPRs stimulation promoted gp91phox and p67phox expression as well the direct interaction between GTP-Rac1 and p67phox in fibroblast cells. Finally, we were able to show that C37, a new small molecule inhibiting the structural and functional interaction between FPRs and uPAR, and Selumetinib, a clinically approved MAPKK/ERK inhibitor, blocked FPRs-mediated ROS production in fibroblasts, thus suggesting new therapeutic strategies for the treatment of fibrosis in SSc

Different duplex/quadruplex junctions determine the properties of anti-thrombin aptamers with mixed folding.

Mixed duplex/quadruplex oligonucleotides have attracted great interest as therapeutic targets as well as effective biomedical aptamers. In the case of thrombin-binding aptamer (TBA), the addition of a duplex motif to the G-quadruplex module improves the aptamer resistance to biodegradation and the affinity for thrombin. In particular, the mixed oligonucleotide RE31 is significantly more effective than TBA in anticoagulation experiments and shows a slower disappearance rate in human plasma and blood. In the crystal structure of the complex with thrombin, RE31 adopts an elongated structure in which the duplex and quadruplex regions are perfectly stacked on top of each other, firmly connected by a well-structured junction. The lock-and-key shape complementarity between the TT loops of the G-quadruplex and the protein exosite I gives rise to the basic interaction that stabilizes the complex. However, our data suggest that the duplex motif may have an active role in determining the greater anti-thrombin activity in biological fluids with respect to TBA. This work gives new information on mixed oligonucleotides and highlights the importance of structural data on duplex/quadruplex junctions, which appear to be varied, unpredictable, and fundamental in determining the aptamer functional properties

Several structural motifs cooperate in determining the highly effective anti-thrombin activity of NU172 aptamer

Despite aptamers are very promising alternative to antibodies, very few of them are under clinical trials or are used as drugs. Among them, NU172 is currently in Phase II as anticoagulant in heart disease treatments. It inhibits thrombin activity much more effectively than TBA, the best-known thrombin binding aptamer. The crystal structure of thrombin-NU172 complex reveals a bimodular duplex/quadruplex architecture for the aptamer, which binds thrombin exosite I through a highly complementary surface involving all three loops of the G-quadruplex module. Although the duplex domain does not interact directly with thrombin, the features of the duplex/quadruplex junction and the solution data on two newly designed NU172 mutants indicate that the duplex moiety is important for the optimization of the protein-ligand interaction and for the inhibition of the enzyme activity. Our work discloses the structural features determining the inhibition of thrombin by NU172 and put the basis for the design of mutants with improved properties

Nanoparticle-guided brain drug delivery: Expanding the therapeutic approach to neurodegenerative diseases

Neurodegenerative diseases (NDs) represent a heterogeneous group of aging-related disorders featured by progressive impairment of motor and/or cognitive functions, often accompanied by psychiatric disorders. NDs are denoted as ‘protein misfolding’ diseases or proteinopathies, and are classified according to their known genetic mechanisms and/or the main protein involved in disease onset and progression. Alzheimer’s disease (AD), Parkinson’s disease (PD) and Huntington’s disease (HD) are included under this nosographic umbrella, sharing histopathologically salient features, including deposition of insoluble proteins, activation of glial cells, loss of neuronal cells and synaptic connectivity. To date, there are no effective cures or disease-modifying therapies for these NDs. Several compounds have not shown efficacy in clinical trials, since they generally fail to cross the blood-brain barrier (BBB), a tightly packed layer of endothelial cells that greatly limits the brain internalization of endogenous substances. By engineering materials of a size usually within 1–100 nm, nanotechnology offers an alternative approach for promising and innovative therapeutic solutions in NDs. Nanoparticles can cross the BBB and release active molecules at target sites in the brain, minimizing side effects. This review focuses on the state-of-the-art of nanoengineered delivery systems for brain targeting in the treatment of AD, PD and HD. © 2021 by the authors. Licensee MDPI, Basel, Switzerland

Small cell lung cancer associated with solitary fibrous tumors of the pleura: A case study and literature review

AbstractIntroductionSmall cell lung cancer (SCLC) is the most aggressive type of lung cancer. The surgical treatment is possible only in a few and defined occasions. The association between SCLC and a solitary fibrous tumor of pleura (SFTP) is extremely rare.Case presentationA 56 year-old man had a lung lesion (size 16 mm) FDG-avid (SUV 7.9) within upper lobe of right lung. No lymph adenopathy or other distant lesion were found. The pathological results of FNAB showed the presence of malignant cells inconclusive for a definitive diagnosis. Following thoracotomy, the exploration of pleural cavity showed an unexpected lesion (size. 3 cm) originating from parietal pleura and not radiologically seen. The intraoperative diagnosis was solitary fibrous tumor of the pleura. Then, an upper right lobectomy was achieved. The histological findings of the lung tumor diagnosed to be a SCLC (p-stage: T1N0M0). An adjuvant treatment was started. At 20 months after the procedure, no recurrence was found.ConclusionSurgery as part of multimodality treatment may be indicated in the treatment of SCLC in the early stage (T1N0M0). However, before proceeding to attend tumor resection an exploration of pleural cavity is mandatory in order to exclude any pleural involvement

Post-pneumonectomy broncho-pleural fistula successfully closed by open-window thoracostomy associated with V.A.C. therapy

AbstractBroncho-pleural fistula (BPF), is a dramatic complication that may occur after lung resection. The treatment is challenging due to its high rate of morbidity and mortality. Herein, a case of BPF associated with empyema, occurred in an elderly patient who had undergone to left pneumonectomy for non-small cell lung cancer (NSCLC), is reported. After various treatments including chest drainage and endoscopic procedures, BPF was successfully closed by open-window thoracotomy associated with vacuum assisted closure (V.A.C.) device therapy. The authors conclude that V.A.C. is a convenient and safe measure in the management of empyema with BPF. Moreover, in similar clinical contexts, V.A.C. may be the only option available that may assure the survival of the patient and the avoiding any later-phases of residual cavity

Tailoring haemophilia A prophylaxis with BAY 81-8973: A case series

BAY 81-8973 is an unmodified, full-length third generation recombinant factor VIII (rFVIII) which offers a more favorable pharmacokinetic (PK) profile, compared to its predecessor sucrose-formulated rFVIII (rFVIII-FS). We here report on a retrospective case series of nine patients affected by hemophilia A (HA), with variable disease severity, bleeding phenotype and comorbidities, to underline our clinical practice on prophylaxis with a recently introduced standard hall-life recombinant Factor VIII. The current case series highlights how the current clinical management of hemophilia is able to personalize treatment in several specific conditions like concomitant illnesses with thrombotic risk and allergic reactions

Digenic mutational inheritance of the integrin alpha 7 and the myosin heavy chain 7B genes causes congenital myopathy with left ventricular non-compact cardiomyopathy

BACKGROUND: We report an Italian family in which the proband showed a severe phenotype characterized by the association of congenital fiber type disproportion (CFTD) with a left ventricular non-compaction cardiomyopathy (LVNC). This study was focused on the identification of the responsible gene/s. METHODS AND RESULTS: Using the whole-exome sequencing approach, we identified the proband homozygous missense mutations in two genes, the myosin heavy chain 7B (MYH7B) and the integrin alpha 7 (ITGA7). Both genes are expressed in heart and muscle tissues, and both mutations were predicted to be deleterious and were not found in the healthy population. The R890C mutation in the MYH7B gene segregated with the LVNC phenotype in the examined family. It was also found in one unrelated patient affected by LVNC, confirming a causative role in cardiomyopathy. The E882K mutation in the ITGA7 gene, a key component of the basal lamina of muscle fibers, was found only in the proband, suggesting a role in CFTD. CONCLUSIONS: This study identifies two novel disease genes. Mutation in MYH7B causes a classical LVNC phenotype, whereas mutation in ITGA7 causes CFTD. Both phenotypes represent alterations of skeletal and cardiac muscle maturation and are usually not severe. The severe phenotype of the proband is most likely due to a synergic effect of these two mutations. This study provides new insights into the genetics underlying Mendelian traits and demonstrates a role for digenic inheritance in complex phenotypes