Reactome Pathway Analysis From Whole-Blood Transcriptome Reveals Unique Characteristics of Systemic Sclerosis Patients at the Preclinical Stage

OBJECTIVE: This study aims to characterize differential expressed pathways (DEP) in subjects with preclinical systemic sclerosis (PreSSc) characterized uniquely by Raynaud phenomenon, specific autoantibodies, and/or capillaroscopy positive for scleroderma pattern. METHODS: Whole-blood samples from 33 PreSSc with clinical prospective data (baseline and after 4 years of follow-up) and 16 matched healthy controls (HC) were analyzed for global gene expression transcriptome analysis via RNA sequencing. Functional Analysis of Individual Microarray Expression method annotated Reactome individualized pathways. ANOVA analysis identified DEP whose predictive capability were tested in logistic regression models after extensive internal validation. RESULTS: At 4 years, 42.4% subjects progressed (evolving PreSSc), while the others kept stable PreSSc clinical features (stable PreSSc). At baseline, out of 831 pathways, 541 DEP were significant at a false discovery rate CONCLUSION: PreSSc had distinguished Reactome pathway signatures compared to HC. Progression to definite SSc was characterized by a shift in biological fingertips. Calcium-related events promoting endothelial damage and vasculopathy may be relevant to disease progression

Macular Microcysts in Mitochondrial Optic Neuropathies: Prevalence and Retinal Layer Thickness Measurements.

PurposeTo investigate the thickness of the retinal layers and to assess the prevalence of macular microcysts (MM) in the inner nuclear layer (INL) of patients with mitochondrial optic neuropathies (MON).MethodsAll patients with molecularly confirmed MON, i.e. Leber's Hereditary Optic Neuropathy (LHON) and Dominant Optic Atrophy (DOA), referred between 2010 and 2012 were enrolled. Eight patients with MM were compared with two control groups: MON patients without MM matched by age, peripapillary retinal nerve fiber layer (RNFL) thickness, and visual acuity, as well as age-matched controls. Retinal segmentation was performed using specific Optical coherence tomography (OCT) software (Carl Zeiss Meditec). Macular segmentation thickness values of the three groups were compared by one-way analysis of variance with Bonferroni post hoc corrections.ResultsMM were identified in 5/90 (5.6%) patients with LHON and 3/58 (5.2%) with DOA. The INL was thicker in patients with MON compared to controls regardless of the presence of MM [133.1±7μm vs 122.3±9μm in MM patients (p<0.01) and 128.5±8μm vs. 122.3±9μm in no-MM patients (p<0.05)], however the outer nuclear layer (ONL) was thicker in patients with MM (101.4±1mμ) compared to patients without MM [77.5±8mμ (p<0.001)] and controls [78.4±7mμ (p<0.001)]. ONL thickness did not significantly differ between patients without MM and controls.ConclusionThe prevalence of MM in MON is low (5-6%), but associated with ONL thickening. We speculate that in MON patients with MM, vitreo-retinal traction contributes to the thickening of ONL as well as to the production of cystic spaces

Reactome pathway analysis from whole-blood transcriptome reveals unique characteristics of systemic sclerosis patients at the preclinical stage

ObjectiveThis study aims to characterize differential expressed pathways (DEP) in subjects with preclinical systemic sclerosis (PreSSc) characterized uniquely by Raynaud phenomenon, specific autoantibodies, and/or capillaroscopy positive for scleroderma pattern.MethodsWhole-blood samples from 33 PreSSc with clinical prospective data (baseline and after 4 years of follow-up) and 16 matched healthy controls (HC) were analyzed for global gene expression transcriptome analysis via RNA sequencing. Functional Analysis of Individual Microarray Expression method annotated Reactome individualized pathways. ANOVA analysis identified DEP whose predictive capability were tested in logistic regression models after extensive internal validation.ResultsAt 4 years, 42.4% subjects progressed (evolving PreSSc), while the others kept stable PreSSc clinical features (stable PreSSc). At baseline, out of 831 pathways, 541 DEP were significant at a false discovery rate <0.05, differentiating PreSSc versus HC with an AUROC = 0.792 ± 0.242 in regression models. Four clinical groups were identified via unsupervised clustering (HC, HC and PreSSc with HC-like features, PreSSc and HC with PreSSc-like features, and PreSSc). Biological signatures changed with disease progression while remaining unchanged in stable subjects. The magnitude of change was related to the baseline cluster, yet no DEP at baseline was predictive of progression. Disease progression was mostly related to changes in signal transduction pathways especially linked to calcium-related events and inositol 1,4,5-triphosphate metabolism.ConclusionPreSSc had distinguished Reactome pathway signatures compared to HC. Progression to definite SSc was characterized by a shift in biological fingertips. Calcium-related events promoting endothelial damage and vasculopathy may be relevant to disease progression

Fondamenti di Telecomunicazioni

Il testo del Couch, ormai un classico nel mondo universitario anglosassone, rappresenta quanto di più vicino si possa oggi immaginare al concetto di libro fondamentale per il settore dei sistemi di telecomunicazione. Esso presenta una scelta di argomenti a spettro molto ampio, e tiene la trattazione a un livello sufficientemente approfondito sui concetti fondamentali, senza tuttavia trascurare gli indispensabili aspetti applicativi. Questa impo- stazione, con l’ausilio di numerosi esempi svolti ed esercizi proposti, rende il questo libro particolarmente adatto come supporto didattico per i corsi delle Lauree triennali dei nuo- vi ordinamenti nazionali nei settori dell’Ingegneria e delle Scienze dell’informazione. Articolando l’insegnamento su due moduli di 6 crediti ciascuno, la suddivisione ideale degli argomenti prevede l’uso dei primi cinque capitoli nel primo modulo, e dei restanti tre nel secondo. Parti specifiche del libro sono inoltre adatte a servire corsi di 4-6 crediti a livello di Lauree specialistiche nel settore dell’Ingegneria e delle Scienze dell’Informazione, al di fuori dell’Ingegneria delle Telecomunicazioni. Il lavoro di traduzione dall’originale in lingua inglese è stato estremamente piacevole, perché ci ha dato l’occasione di rincontrare molti vecchi amici, cioè concetti di base e applicazioni, che magari, travolti nella vita professionale dall’urgenza del contingente, si ha sempre meno il tempo di frequentare. Ma ha richiesto anche molta fatica: da una parte, per il consueto problema della traduzione di rendere chiaramente in italiano concetti e frasi strutturati in un’altra lingua, e dall’altra per la necessità di riadattare, anche integralmente, quelle parti del testo che, specialmente nella descrizione delle applicazioni (reti cellulari, televisione), sono riferite specificamente alla realtà nordamericana e quindi appaiono di scarso interesse per il lettore di lingua italiana. Il nostro approccio è stato quel- lo di evitare il più possibile l’uso del gergo tecnico “italiese”, e cioè di tradurre in lingua italiana il più possibile anche i termini tecnici, amenoché l’equivalente italiano sia di scar- so uso, oppure obsoleto, o un po’ ridicolo. Non vediamo cioè il motivo per cui si debba, in italiano, usare ad esempio la parola inglese “frame” quando l’equivalente “trama” de- scrive esattamente lo stesso concetto, così come non ha senso provare a tradurre l’inglese “clock” universalmente utilizzato anche in Italia con un problematico “orologio” (!?). Per concludere questa dissertazione linguistica, osserviamo che nel mondo accademico è molto diffusa la traduzione di “digital” con “numerico”. Dopo aver seguito questa tendenza per anni, l’abbiamo anche noi abbandonata, passando al meno elegante “digitale” a causa della enorme diffusione del termine anche a livello non tecnico. In fin dei conti, co- me ci insegna N. Negroponte, non siamo oggigiorno tutti quanti “digitali” ? Un caloroso grazie va a Luca Giugno e Cosimo Saccomando per l’opera di correzione del manoscritto e per le relative osservazioni sulla traduzione. Lo staff di Apogeo (il nostro editor A. Kratter Thaler e Stefano Fabiano) si è dimostrato impareggiabile nel guidarci in maniera ottimale al compimento del lavoro senza mai darci l’impressione di tenerci il fiato sul collo..

Enhanced tissue factor expression by blood eosinophils from patients with hypereosinophilia: a possible link with thrombosis.

Thrombotic risk is increased in eosinophil-mediated disorders, and several hypotheses have been proposed to link eosinophilia and thrombosis. In particular, eosinophils have been described as source of tissue factor (TF), the main initiator of blood coagulation; however, this aspect is still controversial. This study was aimed to evaluate whether TF expression varies in eosinophils isolated from normal subjects and patients with different hypereosinophilic conditions. Eosinophils were immunologically purified from peripheral blood samples of 9 patients with different hypereosinophilic conditions and 9 normal subjects. Western blot analysis and real-time polymerase chain reaction (RT-PCR) were performed to test eosinophil TF expression. For comparison, TF expression was evaluated in monocytes from blood donors and in human endothelial (ECV304) and fibroblast (IMR90) cell lines. Western blot analysis revealed a major band of 47,000 corresponding to native TF in homogenates of purified eosinophils with a higher intensity in the 9 patients than in the 9 controls (p<0.0001). According to RT-PCR cycle threshold (Ct), TF gene expression was higher in eosinophils from patients than in those from controls, median (range) 35.10 (19.45-36.50) vs 37.17 (35.33-37.87) (p = 0.002), and was particularly abundant in one patient with idiopathic hypereosinophilic syndrome and ischemic heart attacks (Ct: 19.45). TF gene expression was moderate in monocytes, Ct: 31.32 (29.82-33.49) and abundant in endothelial cells, Ct: 28.70 (27.79-29.57) and fibroblasts, Ct: 22.77 (19.22-25.05). Our results indicate that human blood eosinophils contain variable amounts of TF. The higher TF expression in patients with hypereosinophilic disorders may contribute to increase the thrombotic risk

Representative cytocentrifuge smears of two high purity eosinophil preparations obtained from peripheral blood samples.

<p>May-Grünwald-Giemsa staining, original magnification: X 400 in the upper panels and X 1000 (immersion) in the lower panels.</p

Panel A shows the western blot analysis of tissue factor (TF) in homogenate samples of purified eosinophils from 9 patients with hyperosinophilic conditions (HE) (top), purified eosinophils from 9 normal controls (N) (middle), and purified monocytes from 2 normal controls (M), purified endothelial cells from 2 samples of cell line ECV304 and purified fibroblasts from 4 samples of cell line IMR90 (bottom).

<p>A major band with Mr of 47,000 corresponding to the native TF was found in the eosinophil homogenates from the 9 patients and the 9 controls, with a higher intensity in the former than in the latter. The intensity of the TF band was weaker in monocytes (M1, M2) than in endothelial cells (ECV304) and in fibroblasts (IMR90). Panel B shows the western blot analysis of the ubiquitary protein beta-actin, which was well represented in all patients, normal subjects and positive controls.</p

Demographic and clinical characteristics of patients with hypereosinophilia.

<p>Demographic and clinical characteristics of patients with hypereosinophilia.</p

IgG and IgE Autoantibodies to IgE Receptors in Chronic Spontaneous Urticaria and Their Role in the Response to Omalizumab

Background: Chronic spontaneous urticaria (CSU) is defined as the recurrence of unprovoked transient wheals and itch for more than 6 weeks. Currently, there is an unmet need concerning response prediction in CSU. The present study investigated biomarkers of type I and type IIb autoimmunity as potential predictors of response to omalizumab in CSU. Materials and methods: Differences in levels of IgG and IgE autoantibodies targeting the high- and low-affinity IgE receptors (FcεRI and FcεRII, respectively), as well as spontaneous and specifically triggered leukotriene C (LTC)4 release by basophils from the investigated subjects, were evaluated in 18 consecutive, prospectively enrolled CSU patients and 18 age- and sex-matched, healthy non-atopic controls. Results: The patients with CSU had higher levels of anti-FcεRI IgE (542 (386.25–776.5) vs. 375 (355–418), optical density (OD), p = 0.008), and IgG (297 (214.5–431.25) vs. 193.5 (118–275) OD, p = 0.004) autoantibodies relative to the controls. Simultaneous anti-FcεRI IgG and IgE positivity (i.e., both autoantibody levels above the respective cut-offs) was recorded only in late- and non-responders (3/8 and 1/2, respectively). Discussion: Significantly higher anti-FcεRI IgE autoantibody levels were found in the CSU patients as compared to the controls, supporting FcεRI as an autoallergic target of IgE (autoallergen) in the complex pathophysiological scenario of CSU. The co-occurrence of anti-FcεRI IgG and IgE autoantibodies was documented only in late- and non-responders, but not in early ones, crediting the co-existence of autoimmune and autoallergic mechanisms as a driver of late/poor response to omalizumab

Binding of human recombinant tissue factor (TF) by three commercial anti-TF antibodies (2K1, 4G4 and GMA) evaluated by western blot (upper panel) and enzyme immunoassay (lower panel) methods.

<p>Only 2K1 efficiently recognizes TF with both methods. The last lane of western blotting refers to the size markers (SM). In enzyme immunoassay experiments, data represent the mean of three different measurements.</p