Endothelial-to-mesenchymal transition in systemic sclerosis.

Systemic sclerosis (SSc) is an autoimmune disease characterized by significant vascular alterations and multi-organ fibrosis. Microvascular alterations are the first event of SSc and injured endothelial cells (ECs) may transdifferentiate towards myofibroblasts, the cells responsible for fibrosis and collagen deposition. This process is identified as endothelial-to-mesenchymal transition (EndMT), and understanding of its development is pivotal to identify early pathogenetic events and new therapeutic targets for SSc. In this review, we have highlighted the molecular mechanisms of EndMT and summarize the evidence of the role played by EndMT during the development of progressive fibrosis in SSc, also exploring the possible therapeutic role of its inhibition.

Correction to: Inert gas narcosis in scuba diving, different gases different reactions.

The original version of this article unfortunately contained a mistake.

Inert gas narcosis in scuba diving, different gases different reactions.

PURPOSE: Underwater divers face several potential neurological hazards when breathing compressed gas mixtures including nitrogen narcosis which can impact diver's safety. Various human studies have clearly demonstrated brain impairment due to nitrogen narcosis in divers at 4 ATA using critical flicker fusion frequency (CFFF) as a cortical performance indicator. However, recently some authors have proposed a probable adaptive phenomenon during repetitive exposure to high nitrogen pressure in rats, where they found a reversal effect on dopamine release. METHODS: Sixty experienced divers breathing Air, Trimix or Heliox, were studied during an open water dive to a depth of 6 ATA with a square profile testing CFFF measurement before (T0), during the dive upon arriving at the bottom (6 ATA) (T1), 20 min of bottom time (T2), and at 5 m (1.5 ATA) (T3). RESULTS: CFFF results showed a slight increase in alertness and arousal during the deep dive regardless of the gas mixture breathed. The percent change in CFFF values at T1 and T2 differed among the three groups being lower in the air group than in the other groups. All CFFF values returned to basal values 5 min before the final ascent at 5 m (T3), but the Trimix measurements were still slightly better than those at T0. CONCLUSIONS: Our results highlight that nitrogen and oxygen alone and in combination can produce neuronal excitability or depression in a dose-related response.

H-ferritin and proinflammatory cytokines are increased in the bone marrow of patients affected by macrophage activation syndrome.

Macrophage activation syndrome (MAS) is hyperinflammatory life-threatening syndrome, associated typically with high levels of serum ferritin. This is an iron storage protein including heavy (H) and light (L) subunits, categorized on their molecular weight. The H-/L subunits ratio may be different in tissues, depending on the specific tissue and pathophysiological status. In this study, we analysed the bone marrow (BM) biopsies of adult MAS patients to assess the presence of: (i) H-ferritin and L-ferritin; (ii) CD68+ /H-ferritin+ and CD68+ /L-ferritin+ ; and (iii) interleukin (IL)-1ß, tumour necrosis factor (TNF) and interferon (IFN)-γ. We also explored possible correlations of these results with clinical data. H-ferritin, IL-1ß, TNF and IFN-γ were increased significantly in MAS. Furthermore, an increased number of CD68+ /H-ferritin+ cells and an infiltrate of cells co-expressing H-ferritin and IL-12, suggesting an infiltrate of M1 macrophages, were observed. H-ferritin levels and CD68+ /H-ferritin+ cells were correlated with haematological involvement of the disease, serum ferritin and C-reactive protein. L-ferritin and CD68+ /L-ferritin+ cells did not correlate with these parameters. In conclusion, during MAS, H-ferritin, CD68+ /H-ferritin+ cells and proinflammatory cytokines were increased significantly in the BM inflammatory infiltrate, pointing out a possible vicious pathogenic loop. To date, H-ferritin and CD68+ /H-ferritin+ were associated significantly with haematological involvement of the disease, suggesting biomarkers assessing severity of clinical picture.

Biomarkers and infections in orthopedics: our experience and literature review.

The issue of infections in Orthopedics is rising in importance day by day. In management of Periprosthetic Joint Infection (PJI), classical biomarkers involved in diagnosis process are C-reactive protein (CRP) and Erythrocyte Serum Rate (ESR). Although wide use in PJI diagnosis process, CRP and ESR are not sensitive and specific enough for a correct diagnosis. Orthopedic surgeons, Microbiologists and Infectivologists are very interested in research about new (or rediscovered) biomarkers that can help in the diagnosis of orthopedics infections. In our Institution, a rigid protocol is applied to face suspicious PJI, implemented with LE testing routinely. In our retrospective observational study, we has compared reliability of LE test in relation ICM criteria and the comparison with other diagnostic tests or exams. Our results show that LE is a reliable method especially for intra-operatively PJI diagnosis.

H-ferritin and CD68(+) /H-ferritin(+) monocytes/macrophages are increased in the skin of adult-onset Still's disease patients and correlate with the multi-visceral involvement of the disease.

Adult-onset Still's disease (AOSD) patients may show an evanescent salmon-pink erythema appearing during febrile attacks and reducing without fever. Some patients may experience this eruption for many weeks. During AOSD, exceptionally high serum levels of ferritin may be observed; it is an iron storage protein composed of 24 subunits, heavy (H) subunits and light (L) subunits. The ferritin enriched in L subunits (L-ferritin) and the ferritin enriched in H subunits (H-ferritin) may be observed in different tissues. In this work, we aimed to investigate the skin expression of both H-and L-ferritin and the number of macrophages expressing these molecules from AOSD patients with persistent cutaneous lesions. We observed an increased expression of H-ferritin in the skin, associated with an infiltrate in the biopsies obtained from persistent cutaneous lesions of AOSD patients. Furthermore, a positive correlation between H-ferritin skin levels as well as the number of CD68(+) /H-ferritin(+) cells and the multi-visceral involvement of the disease was observed. Our data showed an increased expression of H-ferritin in the skin of AOSD patients, associated with a strong infiltrate of CD68(+) /H-ferritin(+) cells. Furthermore, a correlation between the levels of H-ferritin as well as of the number of CD68(+) /H-ferritin(+) cells and the multi-visceral involvement of the disease was observed.

Interleukin (IL)-17-producing pathogenic T lymphocytes co-express CD20 and are depleted by rituximab in primary Sjögren's syndrome: a pilot study.

Compelling evidence suggests that interleukin (IL)-17 and IL-17-producing cells play a pivotal role in the pathogenesis of primary Sjögren's syndrome (pSS). We investigated phenotypical and functional effects of the anti-CD20 antibody rituximab (RTX) on circulating and glandular IL-17-producing T cells in pSS. RTX is able to deplete glandular IL-17(+) CD3(+) CD4(-) CD8(-) double-negative (DN) and CD4(+) Th17 cells as well as circulating IL-17(+) DN T cells. A fraction of glandular and circulating IL-17(+) DN cells and CD4(+) T helper type 17 (Th17) cells co-expresses CD20 on the cell surface explaining, at least in part, such depletive capacity of RTX. The exposure to RTX does not rescue the in-vitro corticosteroid resistance of IL-17(+) DN T cells. Our results support further the therapeutic role in pSS of RTX that, despite its B cell specificity, appears able to also hamper IL-17-producing T cells in this disease.

The CD68(+)/H-ferritin(+) cells colonize the lymph nodes of the patients with adult onset Still's disease and are associated with increased extracellular level of H-ferritin in the same tissue: correlation with disease severity and implication for pathogenesis.

In this work, we aimed to evaluate the levels of ferritin enriched in H subunits (H-ferritin) and ferritin enriched in L subunits (L-ferritin) and the cells expressing these two molecules in the lymph node (LN) biopsies obtained from adult-onset Still's disease (AOSD) patients, and the possible correlation among these data and the severity of the disease. Ten patients with AOSD underwent LN biopsy. All the samples were stained by immunofluorescence. A statistical analysis was performed to estimate the possible correlation among both H-ferritin and L-ferritin tissue expression and the clinical picture of the disease. Furthermore, the same analysis was performed to evaluate the possible correlation among the number of CD68(+)/H-ferritin(+) or CD68(+)/L-ferritin(+) cells and the clinical picture. Immunofluorescence analysis demonstrated an increased tissue H-ferritin expression in the LNs of AOSD patients. This increased expression correlated with the severity of the disease. An increased number of CD68 macrophages expressing H-ferritin was observed in the LN samples of our patients. Furthermore, we observed that the number of CD68(+)/H-ferritin(+) cells correlated significantly with the severity of the clinical picture. Our data showed an imbalance between the levels of H- and L-ferritin in LNs of AOSD patients and the evidence of an increased number of CD68(+)/H-ferritin(+) cells in the same organs. Furthermore, a correlation among both the tissue H-ferritin levels and the CD68(+)/H-ferritin(+) cells and the clinical picture was observed.

Groin Pain Syndrome Italian Consensus Conference on terminology, clinical evaluation and imaging assessment in groin pain in athlete.

The nomenclature and the lack of consensus of clinical evaluation and imaging assessment in groin pain generate significant confusion in this field. The Groin Pain Syndrome Italian Consensus Conference has been organised in order to prepare a consensus document regarding taxonomy, clinical evaluation and imaging assessment for groin pain. A 1-day Consensus Conference was organised on 5 February 2016, in Milan (Italy). 41 Italian experts with different backgrounds participated in the discussion. A consensus document previously drafted was discussed, eventually modified, and finally approved by all members of the Consensus Conference. Unanimous consensus was reached concerning: (1) taxonomy (2) clinical evaluation and (3) imaging assessment. The synthesis of these 3 points is included in this paper. The Groin Pain Syndrome Italian Consensus Conference reached a consensus on three main points concerning the groin pain syndrome assessment, in an attempt to clarify this challenging medical problem.

Monocytes from patients with rheumatoid arthritis and type 2 diabetes mellitus display an increased production of interleukin (IL)-1ß via the nucleotide-binding domain and leucine-rich repeat containing family pyrin 3(NLRP3)-inflammasome activation: a possible implication for therapeutic decision in these patients.

A better understanding about the mechanisms involved in the pathogenesis of type 2 diabetes mellitus (T2D) showed that inflammatory cytokines such as tumour necrosis factor (TNF) and interleukin (IL)-1ß play a pivotal role, mirroring data largely reported in rheumatoid arthritis (RA). IL-1ß is produced mainly by monocytes (MO), and hyperglycaemia may be able to modulate, in the cytoplasm of these cells, the assembly of a nucleotide-binding domain and leucine-rich repeat containing family pyrin (NLRP3)-inflammosome, a cytosolic multi-protein platform where the inactive pro-IL-1ß is cleaved into active form, via caspase-1 activity. In this paper, we evaluated the production of IL-1 ß and TNF, in peripheral blood MO of patients affected by RA or T2D or both diseases, in order to understand if an alteration of the glucose metabolism may influence their proinflammatory status. Our data showed, after 24 h of incubation with different glucose concentrations, a significantly increased production of IL-1ß and TNF in all evaluated groups when compared with healthy controls. However, a significant increase of IL-1ß secretion by T2D/RA was observed when compared with other groups. The analysis of relative mRNA expression confirmed these data. After 24 h of incubation with different concentrations of glucose, our results showed a significant increase in NLRP3 expression. In this work, an increased production of IL-1ß by MO obtained from patients affected by both RA and T2D via NLRP3-inflammasome activation may suggest a potential IL-1ß targeted therapy in these patients.

Design evolution in total knee replacement: which is the future?

Total knee replacement (TKR) procedures have evolved in the last 40 years to guarantee improvements implants life and an excellent joint function. The goals for the future evolutions are make easier prosthesis implantation and promote precision. The demand for TKR will rise for the life length increase and for the risk factors impact increase. Design evolution in total knee replacement has to satisfy these new necessities: anatomic congruence, range of motion, less material wear and better resistance to the weight bearing and to the stresses. This paper analyzes design evolution, materials development and future purposes in total knee arthroplasty. At the beginning, TKR history is treated; then we compare several prosthetic designs developed during years. At last the paper speak about recent innovations, like CAD (computer aided design) for example, born to reach the most important goal in the future: better TKR design, is the one that better imitate natural knee characteristics, and that is able to integrate it-self with capsule-ligaments and muscle-tendons patient structures.

Arthroscopic treatment of isolated subscapolaris lesions: our experience.

BACKGROUND AND AIM OF THE WORK: Subscapularis tendon lesions, in particular the isolated ones, are often not recognized and undervalued, so in the literature they are described with a variable incidence. Aim of the work is presenting our experience with the short to medium term follow up results of the arthroscopic repair of isolated subscapularis lesions. METHODS: We retrospectively analyzed 311 shoulder arthroscopies performed by a single senior surgeon, from which we have found 10 isolated subscapularis lesions. After the arthroscopic repair of subscapularis tendon the patients have been evaluated with a median follow up of 17.7 months with specific tests for the subscapularis (Napoleon's and lift off tests) and clinical scores (Constant and UCLA scores). RESULTS: We have obtained the tests negativization with an internal rotation level up to D8. The Constant score reached 86.7 with a median improvement of 49.4 points. The UCLA score at the last follow up was 30.8 with a median improvement of 20.1 points. CONCLUSIONS: Isolated subscapularis lesions are uncommon and often they are not correctly diagnosed. Arthroscopy has a decisive role in both the diagnostic and therapeutic side, with good short to medium term results.

Arthroscopic treatment of meniscal tears: with all-inside suture technique: 1 years follow up results.

BACKGROUND: In the last years meniscal repair surgeries have evolved to less invasive all-arthroscopic technique. PURPOSE: To determine the short-term success rate and reoperation rate of all-inside meniscal sutures with a concomitant anterior cruciate ligament reconstruction. METHODS: Eighteen meniscal repairs were performed using an all inside suture technique. A concomitant anterior cruciate ligament reconstruction was done in all patients.The mean follow-up was 14 months (range 12-18). The success rate was determined by the presence of normal or nearly normal parameters of the Henning Classification based on magnetic resonance imaging. RESULTS: Sixteen (89%) of the meniscal repairs had normal or nearly normal characteristics according to Henning Classification. Two patients (11%) required partial arthroscopicmeniscectomy. CONCLUSIONS: Current techniques do not require accessory posteromedial or posterolateral incisions and significantly reduce the incidence of complications and pain associated with more invasive surgery. The short-term evaluation of the meniscal repairs with concomitant anterior cruciate ligament-reconstruction shown good results with satisfaction of the patients.

Mesenchymal stem cells (MSCs) from scleroderma patients (SSc) preserve their immunomodulatory properties although senescent and normally induce T regulatory cells (Tregs) with a functional phenotype: implications for cellular-based therapy.

Systemic sclerosis (SSc) is a chronic disease, with early activation of the immune system. The aim of our work was to address how SSc-mesenchymal stem cells (MSCs), although senescent, might preserve specific immunomodulatory abilities during SSc. MSCs were obtained from 10 SSc patients and 10 healthy controls (HC). Senescence was evaluated by assessing cell cycle, ß-galactosidase (ß-Gal) activity, p21 and p53 expression; doxorubicin was used as acute senescence stimulus to evaluate their ability to react in stressed conditions. Immunomodulatory abilities were studied co-culturing MSCs with peripheral blood mononuclear cells (PBMCs) and CD4(+) cells, in order to establish both their ability to block proliferation in mixed lymphocyte reaction and in regulatory T cells (Tregs) induction. SSc-MSC showed an increase of senescence biomarkers. Eighty per cent of MSCs were in G0-G1 phase, without significant differences between SSc and HC. SSc-MSCs showed an increased positive ß-Gal staining and higher p21 transcript level compared to HC cells. After doxorubicin, ß-Gal staining increased significantly in SSc-MSCs. On the contrary, doxorubicin abolished p21 activation and elicited p53 induction both in SSc- and HC-MSCs. Interleukin (IL)-6 and transforming growth factor (TGF)-ß-related transcripts and their protein levels were significantly higher in SSc-MSCs. The latter maintained their immunosuppressive effect on lymphocyte proliferation and induced a functionally regulatory phenotype on T cells, increasing surface expression of CD69 and restoring the regulatory function which is impaired in SSc. Increased activation of the IL-6 pathway observed in our cells might represent an adaptive mechanism to senescence, but preserving some specific cellular functions, including immunosuppression.

Receptor binding mode and pharmacological characterization of a potent and selective dual CXCR1/CXCR2 non-competitive allosteric inhibitor.

BACKGROUND AND PURPOSE: DF 2156A is a new dual inhibitor of IL-8 receptors CXCR1 and CXCR2 with an optimal pharmacokinetic profile. We characterized its binding mode, molecular mechanism of action and selectivity, and evaluated its therapeutic potential. EXPERIMENTAL APPROACH: The binding mode, molecular mechanism of action and selectivity were investigated using chemotaxis of L1.2 transfectants and human leucocytes, in addition to radioligand and [(35) S]-GTPγS binding approaches. The therapeutic potential of DF 2156A was evaluated in acute (liver ischaemia and reperfusion) and chronic (sponge-induced angiogenesis) experimental models of inflammation. KEY RESULTS: A network of polar interactions stabilized by a direct ionic bond between DF 2156A and Lys(99) on CXCR1 and the non-conserved residue Asp(293) on CXCR2 are the key determinants of DF 2156A binding. DF 2156A acted as a non-competitive allosteric inhibitor blocking the signal transduction leading to chemotaxis without altering the binding affinity of natural ligands. DF 2156A effectively and selectively inhibited CXCR1/CXCR2-mediated chemotaxis of L1.2 transfectants and leucocytes. In a murine model of sponge-induced angiogenesis, DF 2156A reduced leucocyte influx, TNF-α production and neovessel formation. In vitro, DF 2156A prevented proliferation, migration and capillary-like organization of HUVECs in response to human IL-8. In a rat model of liver ischaemia and reperfusion (I/R) injury, DF 2156A decreased PMN and monocyte-macrophage infiltration and associated hepatocellular injury. CONCLUSION AND IMPLICATIONS: DF 2156A is a non-competitive allosteric inhibitor of both IL-8 receptors CXCR1 and CXCR2. It prevented experimental angiogenesis and hepatic I/R injury in vivo and, therefore, has therapeutic potential for acute and chronic inflammatory diseases.

Recommendations for the management of urinary disorders in multiple sclerosis: a consensus of the Italian Multiple Sclerosis Study Group.

Urinary disorders are uncommon in the initial phases of multiple sclerosis, but increase in frequency as the disease progresses, with a negative impact on quality of life. The goal of this study was to propose a protocol for the diagnosis and treatment of urinary disorders in multiple sclerosis, based on data from the scientific literature and the experience of Italian clinical centres. In particular, the following clinical aspects were considered: what to do with patients with asymptomatic multiple sclerosis; what to do with symptomatic patients; how and when to perform a second-level diagnostic evaluation; and how to treat urinary disorders. A diagnostic-therapeutic algorithm is proposed, that can be applied in Italian clinical centres.

Risk factors for chronic pelvic pain in a cohort of primipara and secondipara at one year after delivery: association of chronic pelvic pain with autoimmune pathologies.

AIM: Over genetic and obsteteric factors, also autoimmunity may be involved in female chronic pelvic pain (CPP) pathogenesis. Our study aims to determinate the prevalence of CPP after one year from delivery, and to investigate the possible influence on CPP of concomitant autoimmune conditions. Methods. We selected a cohort of caucasian primipara and secondipara who delivered in our clinic in 2006. We collected personal, clinical and obstetric data, and asked them about pelviperineal painful symptoms. Results. Mean maternal age is 35.52 years (±4.70), 27.65% of women delivered by cesarean section, 61.04% spontaneously and 11.32% by operative assistance, with partoanalgesia in 10.39% of cases, episiotomy in 41.19%, vaginoperineal tears in respectively 14.10% I degree, 11.13% II degree and 0.93% III-IV degree; 43.60% of women have ever undergone abdominopelvic surgery, 32.84% by laparotomy-laparoscopy, 7.05% by hysteroscopy, 5.01% limited to perineum. Chronic autoimmune diseases affect 78.48% of women, allergies 7.79%, rheumatic pathologies 1.3%, autoimmune endocrinopathies 71.8%; 26.53% of women report pelviperineal painful symptoms, being already present in 2.23% of cases, 12.43% generalised pelvic pain, 4.27% bladder pain, 2.60% vulvodynia, 17. 07% dyspareunia. By monovariate analysis CPP results influenced by III-IV degree vaginoperineal tears, operative assistance, preexisting CPP, previous and actual urinary incontinence, previous abdominopelvic surgical interventions and chronic rheumatic pathologies. Furthermore, rheumatic disease, operative assistance and previous CPP are predictive factors for CPP in the postpartum (AUC=58.10%). Conclusion. Delivery may highlight CPP symptoms in predisposed women affected by chronic autoimmune pathologies.

Clean intermittent self-catheterization in neuro-urology.

Neurogenic lower urinary tract dysfunction (NLUTD) is commonly encountered in rehabilitation settings, and is caused by a variety of pathologies. The management of spinal cord injury (SCI) has been the model of reference for the management of other pathologies with NLUTD. The introduction of intermittent catheterization (IC) led to decline in renal related mortality in SCI patients and allowed an improvement of quality of life (QoL) in all neurogenic patients with NLUTD. IC could be sterile, aseptic or clean. Sterile intermittent catheterization (SIC) is the preferred method of bladder drainage in emergency medicine units and during spinal shock in SCI patients, but it is costly and time-consuming. Catheterizations performed in institutions, such as rehabilitation hospitals and nursing homes, are done aseptically. Clean intermittent catheterization (CIC), i.e. self-catheterization (CISC) or third party catheterization, represents the "gold standard" method for bladder emptying in all neuropathic patients with NLUTD: the technique is safe and effective and results in improved kidney and upper urinary tract status, lessening of vesico-ureteral reflux and amelioration of urinary continence. CISC is mandatory in patients with NLUTD secondary to detrusor areflexia/hypocontractility and in patients suffering from neurogenic detrusor overactivity with detrusor external sphincter dyssynergia and high post void residual of urine, often in combination with antimuscarinics/bladder relaxants. The review summarizes the most important aspects of IC and CISC. Attention was focused on the history of urethral catheterization, aims, materials, advantages, indications, and present-day techniques of CISC, emphasizing the importance of teaching in order to perform correctly the catheterization technique.

[Prevalence of urinary incontinence and pelviperineal rehabilitation during the postpartum in a cohort of primipara and secondipara patients]. / Prevalenza dell'incontinenza urinaria e della riabilitazione pelvi-perineale nel postpartum in una coorte di pazienti primipare e secondipare.

AIM: Urinary incontinence is a classical sign of childbirth-related perineal trauma, the prevalence of which is estimated, depending on age, at between 10% and 50%. Pelviperineal rehabilitation offers an excellent opportunity to prevent pelviperineal disorders. The aim of the present study was to evaluate the prevalence of urinary incontinence and pelviperineal disorders in the local context, and also that of pelviperineal rehabilitation. METHODS: Of the 1793 women who gave birth in Udine in 2006, 900 primipara and secondipara Caucasians with single term pregnancies dated ultrasonically to be within 20 weeks of gestation, were selected. A total of 602 of them were contacted by telephone and two questionnaires were administered. RESULTS: The prevalence of pelviperineal rehabilitation in our population was 4.49%, while that of pelviperineal disorders was much higher, at around 40.20%. The prevalence of urinary incontinence was due in 27.57% of cases to stress and in 14.45% to urgency, with an overlap in 9.8% of cases and a story of prior incontinence in 9.97%. The prevalence of urinary urgency in women subjected to rehabilitation is significantly lower than in those not treated (P=0.004). Dyspareunia represents 16.11% of cases, coital incontinence 0.33%. One case of gas incontinence emerged but there was no case of faecal incontinence. CONCLUSIONS: Pelviperineal rehabilitation in the observed population has a very low prevalence, especially if compared with the high prevalence of disturbances related to dysfunctions of the pelvic floor during postpartum.