Caratterizzazione del fenotipo epidermico del topo knockout per Itch

Itch è una E3 ubiquitina ligasi appartenente alla famiglia HECT, mancante nei topi non-agouti-lethal 18H, anche conosciuti come topi Itchy (Itch-/-), i quali manifestano gravi difetti immunitari e infiammatori e un prurito persistente. Infatti, molti dei suoi substrati sono importanti fattori che regolano la risposta immunitaria e la morte cellulare. Alcuni substrati di Itch sono inoltre fattori di trascrizione coinvolti nello sviluppo epidermico, come p63, cJun, JunB e Notch1. Lo scopo di questo studio è di analizzare il fenotipo epidermico dei topi Itch-/-, sia in condizioni normali sia in seguito a stress fisiologici (irradiazione UV e riparo della ferita). I risultati ottenuti mostrano che l’attività di Itch è importante durante la morfogenesi epidermica, infatti, i topi nascono con un’epidermide più spessa, a causa dell’aumento della proliferazione dei cheratinociti dello strato basale. Inoltre, il differenziamento epidermico è ritardato nei topi Itch-/- appena nati, nonostante la formazione della barriera epidermica rimanga inalterata. Infine, nei topi Itch-/- il processo di riparo della ferita è più accelerato, probabilmente a causa dell’aumento della proliferazione dei cheratinociti. Per tanto i risultati ottenuti suggeriscono che la E3 ubiquitina ligasi Itch ha un ruolo nel controllo di due processi fisiologici: (i) la proliferazione epidermica durante lo sviluppo della cute; (ii) la chiusura della ferita nell’epidermide degli adulti.The HECT-type E3 ubiquitin ligase Itch is absent in the non-agoti-lethal 18H or Itchy mice (Itch-/-), which develop severe immune and inflammatory defects and manifest persistent scratching of the skin. Indeed, among its substrates there are some important in immune response and cell death. A number of Itch targets are also transcriptional factors involved in epidermal formation, such as p63, cJun, JunB, Notch1. The aim of this study is to analyze the epidermal phenotype of Itch-/- mice in normal conditions and under physiological stress (UV irradiation and wound healing). The results obtained show that Itch activity is important during epidermal morphogenesis, indeed mice are born with a thicker epidermis due to an increase of the proliferation rate of the basal layer keratinocytes. In parallel, skin differentiation is delayed in Itch-/- newborn mice, although the formation of the skin barrier remains normal. Interestingly, we have evaluated that Itch-/- mice have an accelerated wound healing process, probably due to an increase in keratinocyte proliferation. All together the results obtained suggest that the E3 ubiquitin ligase Itch has a role in controlling two physiological processes: (i) epidermal proliferation during skin development; (ii) wound closure in adult epidermis

Caratterizzazione del fenotipo epidermico del topo knockout per Itch

Itch è una E3 ubiquitina ligasi appartenente alla famiglia HECT, mancante nei topi non-agouti-lethal 18H, anche conosciuti come topi Itchy (Itch-/-), i quali manifestano gravi difetti immunitari e infiammatori e un prurito persistente. Infatti, molti dei suoi substrati sono importanti fattori che regolano la risposta immunitaria e la morte cellulare. Alcuni substrati di Itch sono inoltre fattori di trascrizione coinvolti nello sviluppo epidermico, come p63, cJun, JunB e Notch1. Lo scopo di questo studio è di analizzare il fenotipo epidermico dei topi Itch-/-, sia in condizioni normali sia in seguito a stress fisiologici (irradiazione UV e riparo della ferita). I risultati ottenuti mostrano che l’attività di Itch è importante durante la morfogenesi epidermica, infatti, i topi nascono con un’epidermide più spessa, a causa dell’aumento della proliferazione dei cheratinociti dello strato basale. Inoltre, il differenziamento epidermico è ritardato nei topi Itch-/- appena nati, nonostante la formazione della barriera epidermica rimanga inalterata. Infine, nei topi Itch-/- il processo di riparo della ferita è più accelerato, probabilmente a causa dell’aumento della proliferazione dei cheratinociti. Per tanto i risultati ottenuti suggeriscono che la E3 ubiquitina ligasi Itch ha un ruolo nel controllo di due processi fisiologici: (i) la proliferazione epidermica durante lo sviluppo della cute; (ii) la chiusura della ferita nell’epidermide degli adulti.The HECT-type E3 ubiquitin ligase Itch is absent in the non-agoti-lethal 18H or Itchy mice (Itch-/-), which develop severe immune and inflammatory defects and manifest persistent scratching of the skin. Indeed, among its substrates there are some important in immune response and cell death. A number of Itch targets are also transcriptional factors involved in epidermal formation, such as p63, cJun, JunB, Notch1. The aim of this study is to analyze the epidermal phenotype of Itch-/- mice in normal conditions and under physiological stress (UV irradiation and wound healing). The results obtained show that Itch activity is important during epidermal morphogenesis, indeed mice are born with a thicker epidermis due to an increase of the proliferation rate of the basal layer keratinocytes. In parallel, skin differentiation is delayed in Itch-/- newborn mice, although the formation of the skin barrier remains normal. Interestingly, we have evaluated that Itch-/- mice have an accelerated wound healing process, probably due to an increase in keratinocyte proliferation. All together the results obtained suggest that the E3 ubiquitin ligase Itch has a role in controlling two physiological processes: (i) epidermal proliferation during skin development; (ii) wound closure in adult epidermis

Gorlin-Goltz syndrome: clinical findings in a Italian population

Gorlin-Goltz syndrome or Nevoid basal cell carcinoma syndrome (NBCCS) is a rare inherited autosomal dominant genodermatosis, with nearly complete penetrance but variable expression. NBCCS results from mutations in the Patched 1 (PTCH1) gene (40%–88% of NBCCS cases with higher estimates closer to 90% in more recent studies). Recently, mutations in suppressor of fused gene (SUFU) and PTCH2 have been found in patients with NBCCS. The estimated prevalence of the disease ranges between 1/57.000 and 1/256.000, with a male-to-female ratio of 1:1. The clinical features arise in the first, second, or third decades of life.1,2 This syndrome includes a wide spectrum of defects encompassing the skin, eyes, central nervous and endocrine system, and bones. Diagnosis is based on fulfilment of: two major diagnostic criteria and one minor diagnostic criterion or one major and three minor diagnostic criteria. Identification of a heterozygous germline PTCH1 or SUFU pathogenic variant on molecular genetic testing establishes the diagnosis if clinical features are inconclusive.3 In this study we sought to investigate clinical aspects in Italian patients with NBCCS. We reviewed all clinical charts of 40 NBCCS patients followed by February 1983 to February 2020 at the “Sapienza” University of Rome, Italy. All patients were investigated in a similar way with periodic evaluations that included dermatological, dental, ophthalmologic, gynecological and cardiological evaluation. Clinical examination included oral inspection, measurement of head circumference and interpupillary distance, examination of the skin for basal cell carcinomas (BCCs), and pits on the palms and soles. Radiographs of the chest, skull, spine, hands, pelvic (female) and teeth panorex were take

Leachate analyses of volcanic ashes from Stromboli volcano: A proxy for the volcanic gas plume composition?

Many volcanoes show a change in chemical composition of the gas phase prior to periods of eruptive activity. Fine‐grained tephra erupted from active vents and transported through volcanic plumes can adsorb, and therefore rapidly scavenge, volatile elements such as sulfur, halogens, and metal species in the form of soluble salts adhering to ash surfaces. Analysis of such water‐soluble surface materials is a suitable supplement for remote monitoring of volcanic gases at inaccessible volcanoes. In this work, ash samples of the 2004 to 2009 eruptive activity of Stromboli volcano were sampled, leached, and analyzed for major and trace elements. Data analysis and interpretation was focused on determining the relationship between chemical composition of water‐soluble components adhering to volcanic ash and the volcano’s activity state. First results show significant temporal variations in ash leachate compositions, reflecting changes in the eruptive style of the volcano. In particular, we reveal that ash leachates S/F and Mg/Na ratios showed marked increases prior to a large‐scale explosion on 15 March 2007.PublishedD172041.2. TTC - Sorveglianza geochimica delle aree vulcaniche attiveJCR Journalreserve

Electrophysiological study of visual pathways in nevoid basal cell carcinoma syndrome patients

Introduction: Gorlin-Goltz syndrome (GGS) also known as nevoid basal cell carcinoma syndrome (NBCCS) is a complex rare genetic disorder characterized by a wide range of clinical and radiological manifestations. Ophthalmological alterations have always been reported, but no study on the eventual pattern visual evoked potentials (pVEPs) abnormalities has yet been published.Purpose: The purpose of the study was to evaluate the functionality of the optic pathways in a group of NBCCS patients through pattern reversal VEPs, after a thorough exclusion of subjects with preexisting ocular and optic pathways pathologies.Methods: Nineteen NBCCS patients (31 eyes) and 20 healthy controls (40 eyes) have been recruited for this study. All subjects underwent an evaluation of the functionality of the optic pathways through pVEPs with small (120'), medium (60'), and large (15') check size stimulation.Results: NBCCS patients showed a statistically significant alteration in the transmission of the macular pathway function when compared to controls. PVEPs analysis confirmed a reduced amplitude and an increased latency of the P100 component, suggesting an involvement of the visual pathway even in the absence of ocular clinical manifestations.Conclusion: Visual pathways may have been affected both by a subclinical myelination deficit, determined directly by the genetic alteration, as well as by neurological abnormalities typical of this syndrome. Further studies are warranted

Neurofibromatosis type 1: ocular electrophysiological and perimetric anomalies

Introduction: Neurofibromatosis type 1 (NF1) is a multisystemic disease caused by the mutation of Nf1 gene located on chromosome 17q11.2. The mutation determines the loss of function of the protein neurofibromin with consequent uncontrolled cellular proliferation. Patients are characterized by a wide range of dermatological, neurological, and ophthalmological symptoms. Purpose: The aim of the study was to evaluate, through pattern visual evoked potentials (p-VEPs) and frequency doubling technology (FDT) Matrix perimetry, the objective and psychophysical functionality of the optic pathways in a group of NF1 patient. Methods: The study group consisted of 26 patients affected by NF1 and 17 healthy controls. Each patient underwent a complete ophthalmological examination, p-VEPs with the evaluation of amplitude and latency of the P100 wave, and FDT perimetry, with the evaluation of central sensitivity (CS), mean deviation (MD), pattern standard deviation (PSD) and glaucoma hemifield test (GHT). Results: NF1 patients showed a statistically significant alteration in the transmission of visual impulse. P-VEPs results highlighted a reduced amplitude and an increased latency of the P100 wave, suggesting an involvement of the visual pathway. Visual field analysis showed a significant reduction in all the observed parameters as well (CS, MD, PSD, and GHT). Conclusion: The present study showed, in NF1 patients, a qualitative and quantitative alteration in the conduction of stimuli through the visual pathways. The observed alterations are present, although, only at a subclinical level. None of the patients included in the study showed any manifest visual deficit nor had any concomitant pathology that might have affected the outcome of the study. In conclusion, electrophysiological exams and computer perimetry may take part, alongside a wider array of exams, in the differential diagnosis and later monitoring of NF1

Validation of the Italian version of the Apathy Evaluation Scale (AES-I) in institutionalized geriatric patients

Objectives: Apathy is a very common symptom in institutionalized elderly and represents a condition of both clinical and public health importance. The Apathy Evaluation Scale (AES) has been shown to be a valid and reliable tool for characterizing, quantifying and differentiating apathy in various health conditions. The aims of this study were to establish the validity and reliability of the Italian version of the AES, and to assess the severity of apathy in a sample of Italian institutionalized geriatric patients.Method: Data were collected from clinical interviews using the AES informant version (AES-I). Associations between measures of apathy and depression, cognitive functioning and perceived quality of life were evaluated, as well as the effect of the living environment on apathetic symptoms.Results: Multiple forms of reliability and validity (i.e. test-retest, internal consistency, discriminability of apathy rating from a standard measure of depression) were satisfied. Our results also show that the characteristics of the care setting may affect the severity of apathetic symptoms.Conclusions: The AES-I Italian version is a reliable and valid instrument for measuring apathy in Italian patients, also allowing a direct comparison with data gathered in other countries.

Inflammatory Response Modulation by Vitamin C in an MPTP Mouse Model of Parkinson's Disease

Vitamin C (Vit C) is anutrient present in many foods, particularly citrus fruits, green vegetables, tomatoes, and potatoes. Vit C is studied for its applications in the prevention and management of different pathologies, including neurodegenerative diseases. Neuroinflammation is a defense mechanism activated by a stimulus or an insult that is aimed at the preservation of the brain by promoting tissue repair and removing cellular debris; however, persistent inflammatory responses are detrimental and may lead to the pathogenesis and progression of neurodegenerative diseases like Parkinson's disease (PD) and Alzheimer's disease. PD is one of the most common chronic progressive neurodegenerative disorders, and oxidative stress is one of the most important factors involved in its pathogenesis and progression.Due to this, research on antioxidant and anti-inflammatory compounds is an important target for counteracting neurodegenerative diseases, including PD. In the central nervous system, the presence of Vit C in the brain is higher than in other body districts, but why and how this occurs is still unknown. In this research, Vit C, with its anti-inflammatory and anti-oxidative properties, is studied to better understand its contribution to brain protection; in particular, we have investigated the neuroprotective effects of Vit C in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced animal model of PD and its role in the modulation of neuroinflammation. First, we observed that Vit C significantly decreased the MPTP-induced loss of tyrosine hydroxylase (TH)-positive dopaminergic neuronal cells in the substantia nigra, as well as microglial cell activation and astrogliosis. Furthermore, gait and spontaneous locomotor activity, evaluated by an automated treadmill and the Open Field test, respectively, were partially ameliorated by Vit C treatment in MPTP-intoxicated animals. In relation to neuroinflammation, results show that Vit C reduced the protein and mRNA expression of inflammatory cytokines such as IL-6, TLR4, TNF-α, iNOS, and CD40, while anti-inflammatory proteins such as IL-10, CD163, TGF-β, and IL-4 increased. Interestingly, we show for the first time that Vit C reduces neuroinflammation by modulating microglial polarization and astrocyte activation. Moreover, Vit C was able to reduce NLRP3 activation, which is linked to the pathogenesis of many inflammatory diseases, including neuroinflammatory disorders. In conclusion, our study provides evidence that Vit C may represent a new promising dietary supplement for the prevention and alleviation of the inflammatory cascade of PD, thus contributing to neuroprotection

Ophthalmic manifestation in neurofibromatosis type 2

Neurofibromatosis type 2 (NF2) is a genetically determined tumor-predisposing syndrome. Ocular manifestations include cataracts, epiretinal membranes, retinal hamartomas, optic disk gliomas, and optic nerve sheath meningiomas. Moreover, optic disk edema, optical atrophy, motility disorders, pupil and lid dysfunction, and neurotrophic keratitis can be observed as indirect signs. An observational study was conducted with the aim to collect clinical data and describe the most frequent NF2 ocular manifestations. Fourteen patients affected by NF2, according to the Manchester criteria, were enrolled. All patients underwent complete ophthalmologic and orthoptic evaluation and a spectral domain optical coherence tomography. Ocular manifestations were present in all patients. The slit lamp evaluation of the anterior segment highlighted cataracts in five patients, keratitis in two patients, corneal leukoma in two patients, and corneal pannus in one patient. Fundus oculi and OCT evaluation identified epiretinal membranes in four patients, vitreoretinal tufts in three patients, optic nerve edema in one patient, and retinal hamartoma in one patient. Moreover, the orthoptic evaluation identified different types of ocular motility disorders in seven patients. This is a descriptive study of a rare disease with poor previous literature. Clinical data are shown, emphasizing the role of NF2-specific ophthalmological and orthoptic findings to help establish an early diagnosis

Increased hexosamine biosynthetic pathway flux alters cell-cell adhesion in INS-1E cells and murine islets

Purpose In type 2 Diabetes, beta-cell failure is caused by loss of cell mass, mostly by apoptosis, but also by simple dysfunction (dedifferentiation, decline of glucose-stimulated insulin secretion). Apoptosis and dysfunction are caused, at least in part, by glucotoxicity, in which increased flux of glucose in the hexosamine biosynthetic pathway plays a role. In this study, we sought to clarify whether increased hexosamine biosynthetic pathway flux affects another important aspect of beta-cell physiology, that is beta-cell-beta-cell homotypic interactions. Methods We used INS-1E cells and murine islets. The expression and cellular distribution of E-cadherin and beta-catenin was evaluated by immunofluorescence, immunohistochemistry and western blot. Cell-cell adhesion was examined by the hanging-drop aggregation assay, islet architecture by isolation and microscopic observation. Results E-cadherin expression was not changed by increased hexosamine biosynthetic pathway flux, however, there was a decrease of cell surface, and an increase in intracellular E-cadherin. Moreover, intracellular E-cadherin delocalized, at least in part, from the Golgi complex to the endoplasmic reticulum. Beta-catenin was found to parallel the E-cadherin redistribution, showing a dislocation from the plasmamembrane to the cytosol. These changes had as a phenotypic consequence a decreased ability of INS-1E to aggregate. Finally, in ex vivo experiments, glucosamine was able to alter islet structure and to decrease surface abundandance of E-cadherin and beta-catenin. Conclusion Increased hexosamine biosynthetic pathway flux alters E-cadherin cellular localization both in INS-1E cells and murine islets and affects cell-cell adhesion and islet morphology. These changes are likely caused by alterations of E-cadherin function, highlighting a new potential target to counteract the consequences of glucotoxicity on beta-cells