NADPH oxidase and mitochondria are relevant sources of superoxide anion in the oxinflammatory response of macrophages exposed to airborne particulate matter

Exposure to ambient air particulate matter (PM) is associated with increased cardiorespiratory morbidity and mortality. In this context, alveolar macrophages exhibit proinflammatory and oxidative responses as a result of the clearance of particles, thus contributing to lung injury. However, the mechanisms linking these pathways are not completely clarified. Therefore, the oxinflammation phenomenon was studied in RAW 264.7 macrophages exposed to Residual Oil Fly Ash (ROFA), a PM surrogate rich in transition metals. While cell viability was not compromised under the experimental conditions, a proinflammatory phenotype was observed in cells incubated with ROFA 100 μg/mL, characterized by increased levels of TNF-α and NO production, together with PM uptake. This inflammatory response seems to precede alterations in redox metabolism, characterized by augmented levels of H2O2, diminished GSH/GSSG ratio, and increased SOD activity. This scenario resulted in increased oxidative damage to phospholipids. Moreover, alterations in mitochondrial respiration were observed following ROFA incubation, such as diminished coupling efficiency and spare respiratory capacity, together with augmented proton leak. These findings were accompanied by a decrease in mitochondrial membrane potential. Finally, NADPH oxidase (NOX) and mitochondria were identified as the main sources of superoxide anion ([Formula presented]) in our model. These results indicate that PM exposure induces direct activation of macrophages, leading to inflammation and increased reactive oxygen species production through NOX and mitochondria, which impairs antioxidant defense and may cause mitochondrial dysfunction

Upregulation of TH/IL-17 Pathway-Related Genes in Human Coronary Endothelial Cells Stimulated with Serum of Patients with Acute Coronary Syndromes

Inflammation plays an essential role in the development and complications of atherosclerosis plaques, including acute coronary syndromes (ACS). Indeed, previous reports have shown that within the coronary circulation of ACS patients, several soluble mediators are released. Moreover, it has been demonstrated that endothelial dysfunction might play an important role in atherosclerosis as well as ACS pathophysiology. However, the mechanisms by which these soluble mediators might affect endothelial functions are still largely unknown. We have evaluated whether soluble mediators contained in serum from coronary circulation of ACS patients might promote changes of gene profile in human coronary endothelial cells (HCAECs)

Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8-associated polyclonal body cavity effusions that mimic primary effusion lymphomas

Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 (KSHV/HHV-8) is involved in the pathogenesis of Kaposi's sarcoma (KS) and 2 lymphoproliferative diseases, primary effusion lymphoma (PEL) and the plasmablastic variant of multicentric Castleman's disease (MCD). PEL is defined as a liquid-phase lymphoma harboring monoclonal rearrangements of immunoglobulin (Ig) variable genes or, more rarely, of T-cell receptor (TCR) genes. As a rule, these elements are used for PEL diagnosis together with the demonstration of HHV-8 infection of the tumor clone. We report here a group of five patients with MCD and KS (n = 2) or MCD only (n = 2) or KS only (n = 1); subjects with KS showed cutaneous and visceral involvement. All 5 patients developed recurrent and massive HHV-8-positive body cavity effusions, often in multiple sites, and died shortly after diagnosis. Cytological examination of effusion specimens disclosed the absence of lymphoma cells in all cases. In the context of MCD or MCD and KS, the effusion cell population was found to include 5–30% of atypical lymphoid cells with plasmacytic features and reminiscent of PEL. Our observations suggest that patients with MCD and/or KS (not exclusively in association with HIV infection) may develop recurrent HHV-8-positive body cavity effusions other than PEL, although with a similar poor outcome. Their common morphologic and molecular features appear to identify a nonlymphomatous entity. The identification of HHV-8 in polyclonal effusions that mimic PEL is notable in light of the association between HHV-8 and PEL, and suggests a putative model of PEL lymphomagenesis

DNA sequence capture and high-throughput sequencing technology: a novel approach to identify a large number of hypertrophic cardiomyopathy-causing genes

Hypertrophic cardiomyopathy (HCM) is the most frequent genetic cardiovascular disease worldwide and is an important cause of heart failure-related disability in young people. To date, more than 20 different genes have been identified and the number is increasing. We evaluted a novel approach to identify causative mutations in a large number of HCM-related and candidate genes. Four HCM patients previously analysed by DHPLC/Sanger sequencing for causative mutations in 8 sarcomeric genes were enrolled in this study. Overall, 234 genes were selected for array on the chip, and a custom sequence capture array was designed for target enrichment of all coding regions. The size of our target was 3,908,196 bp. Each DNA sample was enriched using one custom array, and then sequenced in two runs by the GS FLX System. We obtained an average of 164 Mb/sample, which is equivalent to 503,775 sequencing reads/sample with an average read length of 325.6 bp. Sequence and data analysis were performed using the Roche/454 gsMapper software. High confidence variants were blasted against the SNP database to distinguish between known and unknown variants. We found 7864 different variants, of which 6725 were intronic, 424 intergenic and 715 exonic. About 31% of these variants were novel and 56 novel variants were in 35 HCM related genes. In all patients, we confirmed the mutations and polymorphisms previously identified in them with the DHPLC/Sanger approach. The simultaneous analysis of a vocabulary of genes so to increase sensitivity for the molecular diagnosis may in turn increase the information for those patients in whom traditional screening was not adequate. With this new technology it may be possible to identify mutations in genes that, also acting as phenotype modifiers, could be responsible for clinical variability thereby explaining the pathogenetic mechanism underlying HCM development. Consequently, by reducing time and costs and increasing the sensitivity of molecular testing, routine HCM molecular diagnostics could be implemented also to obtain a model readily applicable to other genetic diseases

DNA Sequence Capture and High Throughput Sequencing Technology: a Novel Approach to Identify a Large Number of Hypertrophic Cardiomyopathy-causing Genes

Hypertrophic cardiomyopathy (HCM) is the most frequent genetic cardiovascular disease worldwide and is an important cause of heart failure-related disability in young people. To date, more than 20 different genes have been identified and the number is increasing. We evaluted a novel approach to identify causative mutations in a large number of HCM-related and candidate genes. Four HCM patients previously analysed by DHPLC/Sanger sequencing for causative mutations in 8 sarcomeric genes were enrolled in this study. Overall, 234 genes were selected for array on the chip, and a custom sequence capture array was designed for target enrichment of all coding regions. The size of our target was 3,908,196 bp. Each DNA sample was enriched using one custom array, and then sequenced in two runs by the GS FLX System. We obtained an average of 164 Mb/sample, which is equivalent to 503,775 sequencing reads/sample with an average read length of 325.6 bp. Sequence and data analysis were performed using the Roche/454 gsMapper software. High confidence variants were blasted against the SNP database to distinguish between known and unknown variants. We found 7864 different variants, of which 6725 were intronic, 424 intergenic and 715 exonic. About 31% of these variants were novel and 56 novel variants were in 35 HCM related genes. In all patients, we confirmed the mutations and polymorphisms previously identified in them with the DHPLC/Sanger approach. The simultaneous analysis of a vocabulary of genes so to increase sensitivity for the molecular diagnosis may in turn increase the information for those patients in whom traditional screening was not adequate. With this new technology it may be possible to identify mutations in genes that, also acting as phenotype modifiers, could be responsible for clinical variability thereby explaining the pathogenetic mechanism underlying HCM development. Consequently, by reducing time and costs and increasing the sensitivity of molecular testing, routine HCM molecular diagnostics could be implemented also to obtain a model readily applicable to other genetic diseases

Genetics of Takotsubo Syndrome

Takotsubo syndrome (TTS) is an enigmatic disease with a multifactorial and still unresolved pathogenesis. A genetic predisposition has been suggested based on the few familial TTS cases. Conflicting results have been published regarding the role of functional polymorphisms in relevant candidate genes, such as α1-, β1-, and β2-adrenergic receptors; G protein–coupled receptor kinase 5; and estrogen receptors. Further research is required to help clarify the role of genetic susceptibility in TTS

Robot-assisted gait training in patients with Parkinson's disease: Implications for clinical practice. A systematic review

none36Background: Gait impairments are common disabling symptoms of Parkinson's disease (PD). Among the approaches for gait rehabilitation, interest in robotic devices has grown in recent years. However, the effectiveness compared to other interventions, the optimum amount of training, the type of device, and which patients might benefit most remains unclear. Objective: To conduct a systematic review about the effects on gait of robot-assisted gait training (RAGT) in PD patients and to provide advice for clinical practice. Methods: A search was performed on PubMed, Scopus, PEDro, Cochrane library,Web of science, and guideline databases,following PRISMA guidelines.We included English articles if they used a robotic system with details about the intervention, the parameters, and the outcome measures. We evaluated the level and quality of evidence. Results: We included twenty papers out of 230 results: two systematic reviews, 9 randomized controlled trials, 4 uncontrolled studies, and 5 descriptive reports. Nine studies used an exoskeleton device and the remainders end-effector robots, with large variability in terms of subjects' disease-related disability. Conclusions: RAGT showed benefits on gait and no adverse events were recorded. However, it does not seem superior to other interventions, except in patients with more severe symptoms and advanced disease.noneCarmignano, Simona Maria; Fundaro', Cira; Bonaiuti, Donatella; Calabro', Rocco Salvatore; Cassio, Anna; Mazzoli, Davide; Bizzarini, Emiliana; Campanini, Isabella; Cerulli, Simona; Chisari, Carmelo; Colombo, Valentina; Dalise, Stefania; Gazzotti, Valeria; Mazzoleni, Daniele; Mazzucchelli, Miryam; Melegari, Corrado; Merlo, Andrea; Stampacchia, Giulia; Boldrini, Paolo; Mazzoleni, Stefano; Posteraro, Federico; Benanti, Paolo; Castelli, Enrico; Draicchio, Francesco; Falabella, Vincenzo; Galeri, Silvia; Gimigliano, Francesca; Grigioni, Mauro; Mazzon, Stefano; Molteni, Franco; Morone, Giovanni; Petrarca, Maurizio; Picelli, Alessandro; Senatore, Michele; Turchetti, Giuseppe; Andrenelli, ElisaCarmignano, Simona Maria; Fundaro', Cira; Bonaiuti, Donatella; Calabro', Rocco Salvatore; Cassio, Anna; Mazzoli, Davide; Bizzarini, Emiliana; Campanini, Isabella; Cerulli, Simona; Chisari, Carmelo; Colombo, Valentina; Dalise, Stefania; Gazzotti, Valeria; Mazzoleni, Daniele; Mazzucchelli, Miryam; Melegari, Corrado; Merlo, Andrea; Stampacchia, Giulia; Boldrini, Paolo; Mazzoleni, Stefano; Posteraro, Federico; Benanti, Paolo; Castelli, Enrico; Draicchio, Francesco; Falabella, Vincenzo; Galeri, Silvia; Gimigliano, Francesca; Grigioni, Mauro; Mazzon, Stefano; Molteni, Franco; Morone, Giovanni; Petrarca, Maurizio; Picelli, Alessandro; Senatore, Michele; Turchetti, Giuseppe; Andrenelli, Elis

Evidence-based improvement of gait in post-stroke patients following robot-assisted training: A systematic review

Background: The recovery of walking after stroke is a priority goal for recovering autonomy. In the last years robotic systems employed for Robotic Assisted Gait Training (RAGT) were developed. However, literature and clinical practice did not offer standardized RAGT protocol or pattern of evaluation scales. Objective: This systematic review aimed to summarize the available evidence on the use of RAGT in post-stroke, following the CICERONE Consensus indications. Methods: The literature search was conducted on Pubmed, Cochrane library and PEDro, including studies with the following criteria: 1) adult post-stroke survivors with gait disability in acute/subacute/chronic phase; 2) RAGT as intervention; 3) any comparators; 4) outcome regarding impairment, activity, and participation; 5) both primary studies and reviews. Results: Sixty-one articles were selected. Data about characteristics of patients, level of disability, robotic devices used, RAGT protocols, outcome measures, and level of evidence were extracted. Conclusion: It is possible to identify robotic devices that are more suitable for specific phase disease and level of disability, but we identified significant variability in dose and protocols. RAGT as an add-on treatment seemed to be prevalent. Further studies are needed to investigate the outcomes achieved as a function of RAGT doses delivered

Gait robot-assisted rehabilitation in persons with spinal cord injury: a scoping review

Background: Many robots are available for gait rehabilitation (BWSTRT and ORET) and their application in persons with SCI allowed an improvement of walking function. Objective: The aim of the study is to compare the effects of different robotic exoskeletons gait training in persons with different SCI level and severity. Methods: Sixty-two studies were included in this systematic review; the study quality was assessed according to GRADE and PEDro's scale. Results: Quality assessment of included studies (n = 62) demonstrated a prevalence of evidence level 2; the quality of the studies was higher for BWSTRT (excellent and good) than for ORET (fair and good). Almost all persons recruited for BWSTRT had an incomplete SCI; both complete and incomplete SCI were recruited for ORET. The SCI lesion level in the persons recruited for BWSTRT are from cervical to sacral; mainly from thoracic to sacral for ORET; a high representation of AIS D lesion resulted both for BWSTRT (30%) and for ORET (45%). The walking performance, tested with 10MWT, 6MWT, TUG and WISCI, improved after exoskeleton training in persons with incomplete SCI lesions, when at least 20 sessions were applied. Persons with complete SCI lesions improved the dexterity in walking with exoskeleton, but did not recover independent walking function; symptoms such as spasticity, pain and cardiovascular endurance improved. Conclusion: Different exoskeletons are available for walking rehabilitation in persons with SCI. The choice about the kind of robotic gait training should be addressed on the basis of the lesion severity and the possible comorbidities