I pazienti con Sindrome di DiGeorge e delezione di Crkl mostrano una sbilanciata risposta proliferativa legata alla riduzione del fattore trascrizionale c-Fos

2012/2013La sindrome di DiGeorge (o del22q11.2) è una immunodeficienza primitiva, avente una incidenza di 1 / 5000 nati vivi. Essa è causata da una delezione, in eterozigosi, di un tratto del braccio lungo del cromosoma 22, di circa 3 MB. La sindrome si accompagna a malformazioni cardiache e del tronco aortico, oltre che ad anomalie del volto e dell’arco palatino, con ipoplasia/aplasia del timo e delle paratiroidi. Oltre alle patologie cardiache gli individui affetti soffrono di tetania e convulsioni, dismorfismi facciali e immunodeficienze con infezioni ripetute sia batteriche che virali, in particolare alle vie aeree sia alte che basse. Spesso sono frequenti episodi di autoimmunità. In generale la sindrome mostra una ampia variabilità fenotipica, anche fra individui della stessa famiglia. Le basi delle alterazioni immunologiche di questa sindrome non sono ancora pienamente comprese e restano anzi assai vaghe. Fra le proteine delete vi è la proteina Crkl, una proteina appartenente alla più ampia famiglia degli adattatori Crk. Crkl prende parte ad importanti processi biologici quali chemotassi, adesione, apoptosi e proliferazione. Crkl è una proteina coinvolta infatti nel signaling di svariati fattori di crescita e citochine come SDF-1α, interferoni di tipo I, GM-CSF ed IL-2. Il nostro scopo è stato quello di comprendere se le alterazioni funzionali dei linfociti T, osservabili in questa sindrome, siano correlabili alla aplo-insufficienza di Crkl. Noi abbiamo analizzato, in un gruppo di pazienti affetti da del22q11.2 la proliferazione, l’apoptosi e l’espressione di vari markers di attivazione, nei linfociti T dopo stimolazione con IL-2 o dopo ingaggio del TCR. Successivamente abbiamo valutato i cambiamenti nei principali fattori chiave della proliferazione, quali le cicline, nonché l’andamento di fattori trascrizionali come c-Fos e stat5, in seguito ad attivazione dei linfociti T. Tutti i pazienti osservati presentavano una evidente riduzione della proteina totale Crkl, cosi come una marcata riduzione della sua forma fosforilata. La fosforilazione di Crkl è indotta principalmente da IL-2 la quale è il principale fattore proliferativo dei linfociti T. IL-2 modula anche l’espressione di Crkl sia a livello di proteina che a livello di mRNA. In aggiunta, in questi soggetti, la proliferazione dei linfociti T dopo triggering del TCR appare anch’essa ridotta, se comparata a soggetti sani di controllo. Noi abbiamo anche osservato un decremento della fosforilazione di stat5 sempre a carico dei linfociti T dei pazienti, dopo stimolazione con IL-2. Stat5 è peraltro noto formare complessi trascrizionali con Crkl i quali complessi sono attivi in sede nucleare nella espressione di geni bersaglio. La successiva analisi dell’andamento dei fattori chiave che regolano il processo proliferativo ha rivelato un decremento nei livelli del fattore trascrizionale c-Fos, facente parte del complesso trascrizionale AP-1 e un calo, variabile nei livelli, della ciclina D3. Questi risultati si sono anche confermati su linee Jurkat dopo silenziamento specifico di Crkl. In conclusione la diminuita proliferazione, benchè solo parziale, osservata nei linfociti T dei pazienti si accompagna a decrementati livelli di Crkl, e fosfo-Crkl con conseguente riduzione anche del fattore trascrizionale c-Fos, e della ciclina D3. Nonché ad una ridotta fosforilazione di stat5, dopo stimolo con IL-2. Il nostro dato suggerisce un ruolo potenziale, ancorché non unico, di Crkl e della sua aploinsufficienza nelle alterazioni funzionali dei linfociti Te in particolare nel deficit proliferativo, in pazienti affetti da del22q11.2.XXVI Ciclo197

Three-axial Fiber Bragg Grating Strain Sensor for Volcano Monitoring

Fiber optic and FBGs sensors have attained a large diffusion in the last years as cost-effective monitoring and diagnostic devices in civil engineering. However, in spite of their potential impact, these instruments have found very limited application in geophysics. In order to study earthquakes and volcanoes, the measurement of crustal deformation is of crucial importance. Stress and strain behaviour is among the best indicators of changes in the activity of volcanoes .. Deep bore-hole dilatometers and strainmeters have been employed for volcano monitoring. These instruments are very sensitive and reliable, but are not cost-effective and their installation requires a large effort. Fiber optic based devices offer low cost, small size, wide frequency band, easier deployment and even the possibility of creating a local network with several sensors linked in an array. We present the realization, installation and first results of a shallow-borehole (8,5 meters depth) three-axial Fiber Bragg Grating (FBG) strain sensor prototype. This sensor has been developed in the framework of the MED-SUV project and installed on Etna volcano, in the facilities of the Serra La Nave astrophysical observatory. The installation siteis about 7 Km South-West of the summit craters, at an elevation of about 1740 m. The main goal of our work is the realization of a three-axial device having a high resolution and accuracy in static and dynamic strain measurements, with special attention to the trade-off among resolution, cost and power consumption. The sensor structure and its read-out system are innovative and offer practical advantages in comparison with traditional strain meters. Here we present data collected during the first five months of operation. In particular, the very clear signals recorded in the occurrence of the Central Italy seismic event of October 30th demonstrate the performances of our device.PublishedWien7TM. Sviluppo e Trasferimento Tecnologic

Transient Decrease of Circulating and Tissular Dendritic Cells in Patients With Mycobacterial Disease and With Partial Dominant IFN\u3b3R1 Deficiency

Interferon-\u3b3 receptor 1 (IFN\u3b3R1) deficiency is one of the inborn errors of IFN-\u3b3 immunity underlying Mendelian Susceptibility to Mycobacterial Disease (MSMD). This molecular circuit plays a crucial role in regulating the interaction between dendritic cells (DCs) and T lymphocytes, thus affecting DCs activation, maturation, and priming of T cells involved in the immune response against intracellular pathogens. We studied a girl who developed at the age of 2.5 years a Mycobacterium avium infection characterized by disseminated necrotizing granulomatous lymphadenitis, and we compared her findings with other patients with the same genetic condition. The patient carried a heterozygous 818del4 mutation in the IFNGR1 gene responsible of autosomal dominant (AD) partial IFN\u3b3R1 deficiency. During the acute infection blood cells immunophenotyping showed a marked reduction in DCs counts, including both myeloid (mDCs) and plasmacytoid (pDCs) subsets, that reversed after successful prolonged antimicrobial therapy. Histology of her abdomen lymph node revealed a profound depletion of tissue pDCs, as compared to other age-matched granulomatous lymphadenitis of mycobacterial origin. Circulating DCs depletion was also observed in another patient with AD partial IFN\u3b3R1 deficiency during mycobacterial infection. To conclude, AD partial IFN\u3b3R1 deficiency can be associated with a transient decrease in both circulating and tissular DCs during acute mycobacterial infection, suggesting that DCs counts monitoring might constitute a useful marker of treatment response

Multisystem autoimmune disease caused by increased STAT3 phosphorylation, and dysregulated gene expression

Signal transducer and activator of transcription (STAT) 3 is a member of the STAT family, and plays a major role in various immunological mechanisms.1 Mutations in STAT3 are associated with a broad spectrum of manifestations, including immunodeficiency, autoimmunity, and malignancy.2 In particular, heterozygous germline loss-of-function (LOF) mutations cause Hyper-IgE syndrome (HIES),3–5 while heterozygous germline gain-of-function (GOF) mutations have recently been associated to multi-organ autoimmune manifestations (i.e. type 1 diabetes, enteropathy, cytopenia, interstitial lung disease, hypothyroidism), lymphoproliferation, short stature, and recurrent infections (OMIM #615952).6–8 We report a 7-year-old boy who presented with early-onset severe enteropathy, and diffuse eczematous dermatitis since birth. During the first weeks of life, Hirschsprung disease was also suspected and surgically treated. Gastrointestinal and cutaneous manifestations were first ascribed to food allergy with quite a good response to amino acid-based formula. In the following months, the patient failed to thrive, and developed respiratory tract infections. At two years, the patient presented with progressive interstitial lung disease characterized by lymphocytic interstitial infiltration leading to pulmonary hypertension, tricuspid insufficiency, and right ventricular heart failure with hepatomegaly. Because of the increased risk of infections, he received intravenous (IV) immunoglobulin infusions (400 mg/kg), prophylaxis with cotrimoxazole and fluconazole. Methylprednisolone at 0.3 mg/kg/day was also given to treat autoimmune manifestations

Tofacitinib May Inhibit Myofibroblast Differentiation from Rheumatoid-Fibroblast-like Synoviocytes Induced by TGF-β and IL-6

During rheumatoid arthritis (RA), the pathogenic role of resident cells within the synovial membrane is suggested, especially for a population frequently referred to as fibroblast-like synoviocytes (FLSs). In this study, we assess the markers of myofibroblast differentiation of RA-FLSs by ex vivo observations and in vitro evaluations following the stimulation with both TGF-β and IL-6. Furthermore, we investigated the possible inhibiting role of tofacitinib, a JAK inhibitor, in this context. Myofibroblast differentiation markers were evaluated on RA synovial tissues by immune-fluorescence or immune-histochemistry. RA-FLSs, stimulated with transforming growth factor (TGF-β) and interleukin-6 (IL-6) with/without tofacitinib, were assessed for myofibroblast differentiation markers expression by qRT-PCR and Western blot. The same markers were evaluated following JAK-1 silencing by siRNA assay. The presence of myofibroblast differentiation markers in RA synovial tissue was significantly higher than healthy controls. Ex vivo, α-SMA was increased, whereas E-Cadherin decreased. In vitro, TGF-β and IL-6 stimulation of RA-FLSs promoted a significant increased mRNA expression of collagen I and α-SMA, whereas E-Cadherin mRNA expression was decreased. In the same conditions, the stimulation with tofacitinib significantly reduced the mRNA expression of collagen I and α-SMA, even if the Western blot did not confirm this finding. JAK-1 gene silencing did not fully prevent the effects of stimulation with TGF-β and IL-6 on these features. TGF-β and IL-6 stimulation may play a role in mediating myofibroblast differentiation from RA-FLSs, promoting collagen I and α-SMA while decreasing E-Cadherin. Following the same stimulation, tofacitinib reduced the increases of both collagen I and α-SMA on RA-FLSs, although further studies are needed to fully evaluate this issue and confirm our results

Plasmacytoid Dendritic Cells Depletion and Elevation of IFN-γ Dependent Chemokines CXCL9 and CXCL10 in Children With Multisystem Inflammatory Syndrome

BackgroundSARS-CoV-2 occurs in the majority of children as COVID-19, without symptoms or with a paucisymptomatic respiratory syndrome, but a small proportion of children develop the systemic Multi Inflammatory Syndrome (MIS-C), characterized by persistent fever and systemic hyperinflammation, with some clinical features resembling Kawasaki Disease (KD).ObjectiveWith this study we aimed to shed new light on the pathogenesis of these two SARS-CoV-2-related clinical manifestations.MethodsWe investigated lymphocyte and dendritic cells subsets, chemokine/cytokine profiles and evaluated the neutrophil activity mediators, myeloperoxidase (MPO), and reactive oxygen species (ROS), in 10 children with COVID-19 and 9 with MIS-C at the time of hospital admission.ResultsPatients with MIS-C showed higher plasma levels of C reactive protein (CRP), MPO, IL-6, and of the pro-inflammatory chemokines CXCL8 and CCL2 than COVID-19 children. In addition, they displayed higher levels of the chemokines CXCL9 and CXCL10, mainly induced by IFN-gamma. By contrast, we detected IFN-alpha in plasma of children with COVID-19, but not in patients with MIS-C. This observation was consistent with the increase of ISG15 and IFIT1 mRNAs in cells of COVID-19 patients, while ISG15 and IFIT1 mRNA were detected in MIS-C at levels comparable to healthy controls. Moreover, quantification of the number of plasmacytoid dendritic cells (pDCs), which constitute the main source of IFN-alpha, showed profound depletion of this subset in MIS-C, but not in COVID-19.ConclusionsOur results show a pattern of immune response which is suggestive of type I interferon activation in COVID-19 children, probably related to a recent interaction with the virus, while in MIS-C the immune response is characterized by elevation of the inflammatory cytokines/chemokines IL-6, CCL2, and CXCL8 and of the chemokines CXCL9 and CXL10, which are markers of an active Th1 type immune response. We believe that these immunological events, together with neutrophil activation, might be crucial in inducing the multisystem and cardiovascular damage observed in MIS-C

The Association between Influenza and Pneumococcal Vaccinations and SARS-Cov-2 Infection : Data from the EPICOVID19 Web-Based Survey

The present study aims to evaluate whether influenza and pneumococcal vaccinations are associated with positive nasopharyngeal swab (NPS) testing to detect SARS-CoV-2. Data from the Italian cross-sectional web-based survey (EPICOVID19), based on a self-selection sample of individuals aged 6518, were considered. The probability of a positive SARS-CoV-2 NPS test result as a function of influenza or anti-pneumococcal vaccination was evaluated using multivariable logistic regression, stratifying analysis by age (<65 years, 6565 years). From April 2020, 170,731 individuals aged <65 years and 28,097 6565 years filled out the EPICOVID19 questionnaire. Influenza and anti-pneumococcal vaccinations were received, respectively, by 16% and 2% of those <65 years, and by 53% and 13% of those 6565 years. SARS-CoV-2 NPS testing was reported by 6680 participants. Anti-pneumococcal and influenza vaccinations were associated with a decreased probability of a SARS-CoV-2 NPS positive test in the younger participants (OR = 0.61, 95% CI 0.41\u20130.91; OR = 0.85, 95%CI 0.74\u20130.98; respectively). A significantly lower probability of a positive test result was detected in the individuals 6565 years who received anti-pneumococcal vaccination (OR = 0.56, 95%CI 0.33\u20130.95). These results need to be confirmed by further investigations, but they are relevant given the probable coexistence of influenza, bacterial infections, and COVID-19 over the coming autumn\u2013winter season

Half versus full vacuum suction drainage after modified radical mastectomy for breast cancer- a prospective randomized clinical trial[ISRCTN24484328]

BACKGROUND: Suction drains are routinely used after modified radical mastectomy and are an important factor contributing to increased hospital stay as the patients are often discharged only after their removal. Amongst various factors that influence the amount of postoperative drainage, the negative suction pressure applied to the drain has been reported to be of great significance. While a high negative suction pressure is expected to drain the collection and reduce the dead space promptly, it may also prevent the leaking lymphatics from closing and lead to increased drainage from the wound. Against this background a prospective randomized clinical study was conducted to compare the amount and duration of drainage between a half negative suction and full vacuum suction drainage in patients following modified radical mastectomy. The associated postoperative morbidity was also compared between the two groups. METHODS: 85 FNAC (fine needle aspiration cytology) proven cases of locally advanced breast cancer were randomized. (Using randomly ordered sealed envelops, which were opened immediately before the closure of the wound) in to 50 patients with full vacuum suction (pressure = 700 g/m2) and 35 cases in to half vacuum suction drainage (pressure = 350 g/m2) groups. The two groups were comparable in respect of age, weight, and technique of operation and extent of axillary dissection. Surgery was performed by the same surgical team comprising of five surgeons (two senior and three resident surgeons) using a standardized technique with electrocautery. External compression dressing was provided over the axilla for first 48 hrs and following that patients were encouraged to do active and passive shoulder exercises. The outcomes measured were postoperative morbidity and the length of hospital stay. Statistical methods used: Descriptive studies were performed with SPSS version 10 and group characteristics were compared using student t-test. RESULTS: Half vacuum suction drains were removed earlier than the full suction vacuum suction drains. There was no significant difference in the incidence of seroma formation in the two groups and there was a significant reduction in the total hospital stay in patients with half vacuum suction drainage systems as compared to the full suction drainage group (p < 0.001) without any added morbidity. CONCLUSIONS: Half negative suction drains provide an effective compromise between no suction and full or high suction drainage after modified radical mastectomy by reducing the hospital stay and the post operative morbidity including post operative seromas

Rehabilitación de un ejemplar de Tortuga verde del Pacífico (Chelonia mydas) por ingesta de plástico y posterior trauma en vida silvestre

Este trabajo tiene como objetivo concientizar sobre el daño que hacen los plásticos vertidos en el mar y proporcionar información acerca del manejo médico de ejemplares de tortugas verdes del pacifico bajo cuidado humano ante la necesidad de atención de los mismos. Se presenta un caso de un ejemplar hembra adulto de Chelonia mydas rescatado en la ciudad de Constitución a 200 Km al sur poniente del Centro de Rehabilitación de Fauna Silvestre (CRFS) de Parque Safari. El ejemplar evidencia un cuadro de síndrome de embollamiento con flotación caudal, debido al acumulo de gases propio del íleo paralitico, a su vez presenta lesiones lacerativas en aleta de miembro anterior izquierdo. Se decide realizar la ruta diagnóstica con examen clínico y métodos complementarios como radiografías, ecografías y análisis sanguíneos, además de un proceso quirúrgico para acelerar la recuperación de heridas. Luego de tres meses de proceso de rehabilitación se concluye con la reinserción del ejemplar en la bahía Chascos, comuna de Caldera, a un total de 800 km al norte del CRFS. Para la rehabilitación de este ejemplar, se recorrió cerca de 1000 km por las rutas de Chile desde su rescate hasta su reinserción, con colaboraciones desde el sector público y privado para la gestión de su proceso