Posterior Tibial Tendon Rupture Associated With Anterolateral Distal Tibial and Medial Malleolar Fracture and a Novel Pattern of Tibiofibular Syndesmotic Injury: A Case Report and Review of the Literature.

A posterior tibial tendon (PTT) rupture associated with ankle fractures is a very rare condition. Ankle pronation and external rotation (PER) movement are the typical traumatic mechanism. This injury is frequently overlooked preoperatively. Early diagnosis and treatment are very important to prevent the serious consequences related to functional PTT insufficiency on biomechanics of the foot. Few cases have been described in the literature that highlight the relationship between PTT rupture and PER type ankle fracture with a medial malleolar fracture. We present a case of a complete PTT rupture in a closed atypical ankle fracture in which a medial malleolar fracture was associated with a very large fragment from the anterolateral distal tibia (Tillaux-Chaput fragment) and a concomitant avulsion fracture from the anteromedial portion of the fibula (Lefort-Wagstaffe fragment), with a novel pattern never described before

Contacts between mast cells and dendritic cells in the human skin

Langerhans cells are a dendritic cell type characteristic of the epidermis. Since mast cells secrete molecules potentially influent on dendritic cell differentiation, we have addressed the degree of proximity between these two cell types in biopsies of skin diseases characterized by massive influx of dendritic cell precursors. By fluorescence microscopy, avidin labeled mast cells were found in contact with CD1a+ dendritic cells. By electron microscopy, contacts between mast cells and blunty dendritic cells were found in areas corresponding to those where CD1a+ cells were localized by immunohistochemistry. We propose that mast cells can induce the differentia­ tion of precursors into Langerhans cells through both the release of short range acting soluble factors and contact­mediating plasma membrane molecules

Conservazione e gestione della naturalità negli ecosistemi marino-costieri. Il trapianto delle praterie di Posidonia oceanica

Dal 1980 ad oggi, ogni 30 minuti, si è persa un’area ricoperta di fanerogame marine equivalente ad 1 campo di calcio cioè, nel tempo di gioco di una partita, 3 campi di fanerogame marine scompaiono”. Nell’area mediterranea, le praterie di Posidonia oceanica, uno degli ecosistemi marini di maggior pregio, sono oggi in forte regressione, principalmente a causa delle diverse pressioni antropiche che insistono sulla fascia costiera. Accanto alle numerose azioni di protezione, si è ultimamente sviluppata l’idea di tutelare le praterie anche mediante interventi di trapianto, in grado sia di accelerare la lenta colonizzazione della prateria sia di favorire il ripristino dell’ecosistema degradato. Nel lavoro “Il trapianto delle praterie di Posidonia oceanica”, ISPRA presenta una rassegna delle informazioni ad oggi disponibili su tale tematica, derivanti sia dall’analisi critica della letteratura scientifica esistente e degli aspetti normativi vigenti, sia da specifiche esperienze tecniche maturate in alcuni casi studio. Tale manuale può inoltre costituire un valido strumento di supporto per tecnici e amministratori coinvolti nella gestione della fascia costiera e nella tutela degli ecosistemi ivi presenti

484. Preclinical Proof of Concept of Transcriptional Silencing and Replacement Strategy for Treatment of Retinitis Pigmentosa Due To RHODOPSIN Mutations

Silencing and replacement strategy is a promising approach to overcome mutational heterogeneity of genetic defects. In autosomal dominant retinitis pigmentosa (adRP) due to rhodopsin gene (RHO) approximately 200 different mutations have been described, posing a challenge for the design of effective therapeutics.We designed a silencing and replacement strategy based on transcriptional silencing through an artificial zinc finger DNA-binding protein lacking effector domains (ZF6DBD), and tested both efficacy and safety in two animal models.In a murine model of adRP, we show that AAV-mediate retinal delivery (AAV2/8-CMV-ZF6-DBD) is associated with selective transcriptional silencing of the human mutated allele resulting in morphological and functional (Electroretinography, ERG a-wave and b-wave responses) rescue. We then tested the effect of transcriptional silencing in the porcine large pre-clinical model. Delivery of a low dose (AAV2/8-CMV-ZF6-DBD, 1×10e10 vector genomes, vg) of the ZF6 transcriptional silencer to the porcine retina resulted in robust transcriptional silencing of the endogenous porcine RHO transcript. Cell sorting of transduced photoreceptors showed an almost complete RHO transcriptional silencing effect (90% RHO transcriptional repression), underscoring the potency of the system. To determine the safety of the zinc-finger silencer we performed extensive RNA-seq analysis on treated and control retinae. The data sets generated demonstrate selective RHO gene transcriptional repression and a remarkably low number of differential expressed genes (DEGs), supporting specificity and thus, safety. The co-administration to the porcine retina of the AAV-ZF6 silencer (AAV2/8-CMV-ZF6-DBD) and the AAV-RHO replacement (5×10e11 vg, AAV2/8-GNAT1-HumanRHO) constructs resulted in a balanced silencing and replacement effect. This data support the use of zinc-finger based RHO transcriptional silencing for the development of a clinical trial for adRP patients

A Different Microbiome Gene Repertoire in the Airways of Cystic Fibrosis Patients with Severe Lung Disease

In recent years, next-generation sequencing (NGS) was employed to decipher the structure and composition of the microbiota of the airways in cystic fibrosis (CF) patients. However, little is still known about the overall gene functions harbored by the resident microbial populations and which specific genes are associated with various stages of CF lung disease. In the present study, we aimed to identify the microbial gene repertoire of CF microbiota in twelve patients with severe and normal/mild lung disease by performing sputum shotgun metagenome sequencing. The abundance of metabolic pathways encoded by microbes inhabiting CF airways was reconstructed from the metagenome. We identified a set of metabolic pathways differently distributed in patients with different pulmonary function; namely, pathways related to bacterial chemotaxis and flagellar assembly, as well as genes encoding efflux-mediated antibiotic resistance mechanisms and virulence-related genes. The results indicated that the microbiome of CF patients with low pulmonary function is enriched in virulence-related genes and in genes encoding efflux-mediated antibiotic resistance mechanisms. Overall, the microbiome of severely affected adults with CF seems to encode different mechanisms for the facilitation of microbial colonization and persistence in the lung, consistent with the characteristics of multidrug-resistant microbial communities that are commonly observed in patients with severe lung disease

320 transcriptional silencing via synthetic dna binding protein lacking canonical repressor domains as a potent tool to generate therapeutics

Transcription factors (TFs) function by the combined activity of their DNA-binding domains (DBDs) and effector domains (EDs). Here we show that in vivo delivery of an engineered DNA-binding protein uncoupled from the repressor domain entails complete and gene-specific transcriptional silencing. To silence RHODOPSIN (RHO) gain-of-function mutations, we engineered a synthetic DNA-binding protein lacking canonical repressor domains and targeted to the regulatory region of the RHO gene. AAV-mediate retinal delivery at a low dose (AAV2/8-CMV-ZF6-DBD, 1×10e10 vector genomes, vg) in the porcine retina resulted in selective transcriptional silencing of RHO expression. The rod photoreceptors (the RHO expressing cells) transduced cells when isolated by FACS-sorting showed the remarkable 90% RHO transcriptional repression. To evaluate genome-wide transcriptional specificity, we analyzed the porcine retina transcriptome by RNA sequencing (RNA-Seq). The differentially expressed genes (DEGs) analysis showed that only 19 genes were perturbed. In this study, we describe a system based on a synthetic DNA binding protein enabling targeted transcriptional silencing of the RHO gene by in vivo gene transfer. The high rate of transcriptional silencing occurring in transduced cells supports applications of this regulatory genomic interference with a synthetic trans-acting factor for diseases requiring gene silencing in a large number of affected cells, including for instance a number of neurodegeneration disorders. The result support a novel mode of gene targeted silencing with a DNA-binding protein lacking intrinsic activity

The diagnostic role of Next Generation Sequencing in uncovering isolated splenomegaly: A case report

Many diseases can induce splenomegaly, however, about 5% of splenomegalies are idiopathic. When there is no underlying treatable cause, and the splenomegaly significantly affects the quality of life, splenectomy is the best therapeutic choice. A 67-year-old woman had idiopathic and asymptomatic splenomegaly. The increase in splenomegaly resulted in hypersplenism with cytopenia and symptoms related to abdominal discomfort. The patient underwent splenectomy which led to clinical improvement. A histological examination showed the presence of hematopoietic tissue. Peripheral blood Next Generation Sequencing with the myeloid panel SOPHiA Genetics showed the following mutations: ASXL1, SRSF2, KRAS and TET2. Three out of these four mutations were also found in the splenic tissue. Next Generation Sequencing could be useful in the diagnosis of splenomegalies associated with myeloproliferative neoplasms otherwise defined as idiopathic, in order to address a therapeutic strategy

Inhalation therapy in the next decade : Determinants of adherence to treatment in asthma and COPD

Peer reviewedPublisher PD

Cellular response to photodynamic therapy in chronic wounds

A wound is considered chronic if it does not heal timely. Basic processes at work are similar to acute healing, but their persistence leads to abundant granulation tissue and possibly fibrosis, scar contraction and loss of function [1]. Wounds may become chronic because of local and systemic conditions and are a major concern in clinical dermatology. Photodinamic therapy (PDT) has been proposed for these cases, through the administration of aminolaevulinic acid (ALA), leading to the synthesis of the photosensitizer protoporphyrin IX (PpIX), followed by illumination with visible light [2]. To study the effects of this therapy on the cell infiltrate of chronic wounds, cryosections of lesions before and after PDT were post fixed in cold acetone and stained with haematoxilin and eosin or immunolabeled with antibodies to laminin 5 (for basement membrane), HSP47 (for fibroblasts), alpha-smooth muscle actin (for myofibroblasts), SPM250 (for granulocytes). Intact neighbor skin was used as control. The cellular infiltrate, as well as the thickness of epidermis, the vascularization and the number of fibroblasts appeared increased in chronic wounds over healthy skin. After completion of PDT, fibroblasts appeared further increased in number, the treatment seemed to stimulate the connective tissue cells responsible for tissue repair rather than the inflammatory infiltrate (Supported by Ente Cassa di Risparmio di Firenze, grant 3681 to S.B.)

Activation of anti-inflammatory cell pathways in skin ulcers upon photodynamic therapy

This study was aimed at assessing the variations in skin histology in ulcers caused by chronic venous insufficiency of the lower extremities, upon photodynamic therapy (PDT). The study was approved by the ethical committee of Azienda Sanitaria Firenze. Patient assessment included clinical history, physical examination and echo-Doppler sonography. Four to five sessions of photodynamic therapy (20% 5-aminolevulinic acid gel application followed by 3 min irradiation at 630 nm, total 180 J/cm2) were administered to 15 patients refractory to previous conventional treatments. Skin biopsies were embedded in freezing tissue medium and quick frozen. Cryosections were post fixed in cold acetone. Sections from each case were stained with hematoxylin and eosin or labelled with primary antibodies against the following antigens: MHC-II class, DC-SIGN, CD68, CD163, BDCA2, CD4, CD25, TNF alpha. In some instances avidin and Ulex europaeus lectin were used to tag mast cells and vessels respectively. Upon treatment, MHC-II signal intensity per positive cell and TNF alpha signal in mast cells increased, as well as the numbers of CD68 positive/CD163 positive cells (M2 macrophages), BDCA2 positive (plasmacytoid dendritic) cells and CD4 positive/CD25 positive (Treg) lymphocytes number. Diffuse tissue TGF beta positivity also increased. DC-SIGN positive cells decreased in number. Mast cells were found in proximity of dendritic cells and of vessels; plasmacytoid dendritic cells were found in proximity of T reg cells. Clinically, mild decrease in ulcer size and granulation at ulcer borders were observed. Therefore treatment apparently led to the activation of cells and of intercellular communication pathways possibly down-regulating the inflammatory response. The same treatment had been shown to increase mast cell expression of basic fibroblast growth factor and fibroblast number (1), potentially responsible for increased production of extracellular matrix. Both types of effects could by synergistically beneficial for ulcer repair. This work was supported by the Italian Ministry of Education, MIUR FIRB 2010 and MIUR PRIN-2009; University and Research, Ente Cassa di Risparmio di Firenze (Grant n. 3681 year 2012) and Foemina Foundation