Single nucleotide polymorphisms in the promoter regions of Foxp3 and ICOSLG genes are associated with Alopecia areata.

Alopecia areata (AA), an autoimmune disease affecting anagen stage hair follicles, is associated with polymorphisms in immune-related genes and with decreased number of CD4+ CD25+ T regulatory cells (Treg). Treg function is modulated by the forkhead box protein 3 (FOXP3) transcription factor and by inducible costimulator (ICOS), through interaction with the relative ligand, ICOSLG, whose genes are polymorphic. The aim of the study was to investigate whether specific single nucleotide polymorphisms (SNPs) of the rs2294020 FOXP3 and/or rs378299 ICOSLG genes may be associated with AA. A case-control study was performed in 120 AA patients and 84 controls. SNPs were analyzed by gene sequencing. FOXP3 and ICOSLG gene expressions were analyzed by real-time PCR. Increased frequencies of the genotype carrying the FOXP3 rs2294020-3675(A) [P = 0.002, OR (95 % CI): 2.55 (1.2-2.7)] or the ICOSLG rs378299-509(C) [P = 0.01, OR (95 % CI): 2.21 (1.1-2.6)] allelic variants were observed in AA patients than in controls. The genotype carrying the combination of the FOXP3 rs2294020-3675(A) and ICOSLG rs378299-509(C) allelic variants with the HLA DQB1*03 allele was more frequently present in AA patients than in controls (P = 0.04). The presence of the FOXP3 rs2294020-3675(A) or the ICOSLG rs378299-509(C) allelic variant was associated with reduced relative gene expression in AA patients. These data suggest that rs2294020 SNP of FOXP3 gene and rs378299 SNP of ICOSLG gene are associated with AA and with a reduced expression of the FOXP3 and ICOSLG genes in alopecia patients.Genetic analysis in patients with Alopecia Areat

Pathophysiology of HCV-Related cryoglobulinemic vasculitis: molecular, immunological and clinical analysis

ABSTRACT INTRODUZIONE: L’infezione da virus dell’epatite C (HCV) rappresenta la principale causa di malattia cronica del fegato, che può evolvere in cirrosi e carcinoma epatocellulare (HCC). L’infezione cronica da HCV è inoltre caratterizzata dall’insorgenza di diverse manifestazioni extraepatiche, la più comune delle quali è la crioglobulinemia mista (MC). Si tratta di una vasculite mediata da immunocomplessi che coinvolge vasi sanguigni di piccolo calibro, caratterizzata da una proliferazione B-cellulare con possibile evoluzione in linfoma non-Hodgkin a cellule B. Il caratteristico tropismo dell’HCV per il tessuto linfoide è stato ampiamente dimostrato. La persistenza dell’HCV può rappresentare uno stimolo continuo per il sistema immunitario dell’ospite. Questo potrebbe sostenere proliferazioni oligo/monoclonali B-cellulari con l’espansione di un singolo clone dominante da cui deriva, ad esempio, la sintesi di IgM monoclonali con attività di fattore reumatoide nella crioglobulinemia mista. D’altro canto, la proliferazione linfocitaria può essere promossa in maniera diretta dall’infezione e dalla replicazione dell’HCV all’interno delle cellule B. Su queste basi, l’approccio terapeutico della MC HCV-associata è attualmente basato sulla terapia antivirale, in presenza o assenza di deplezione B-cellulare con rituximab. Tra i fattori dell’ospite che influenzano la risposta virologica ai regimi terapeutici antivirali basati sull’uso di interferone-α peghilato e ribavirina, i polimorfismi del gene dell’IL28B sono stati identificati mediante studi di associazione genomewide come fattori predittivi attendibili dell’esito della terapia antivirale. Lo scopo dello studio consiste nella valutazione della possibile associazione tra i polimorfismi del singolo nucleotide (SNPs) rs12979860/ rs8099917 a livello del gene IL28B e la risposta terapeutica in pazienti con MC HCV-correlata. È stata anche considerata la possibile associazione dei polimorfismi con le manifestazioni cliniche specifiche della MC nonché il loro possibile ruolo nella valutazione della risposta completa (intesa come virologica, molecolare ed immunologica) alla terapia. METODI: Sono stati reclutati 159 pazienti infetti da HCV con MC e 172 pazienti infetti da HCV senza MC, trattati con terapia antivirale basata su interferone-α peghilato e ribavirina. È stata effettuata la genotipizzazione degli SNPs rs12979860/ rs8099917 in tutti i pazienti. In 106 (66.6%) pazienti affetti da crioglobulinemia mista è stato determinato il profilo clonale delle cellule B circolanti. RISULTATI: Il genotipo T/T IL28B rs12979860 è risultato più frequente nei pazienti con MC piuttosto che nei pazienti senza MC (17% vs 8.1%, p=0.02). Il genotipo C/C è risultato associato con un maggior tasso di risposte complete alla terapia (52.6% vs 39.2%, p=0.13). È stata inoltre riscontrata una maggior frequenza di espansioni clonali B-cellulari in circolo (84.2% vs 55.9%, p=0.005), coinvolgimento renale (21% vs 2.9%, p=0.003) e linfomi non-Hodgkin a cellule B (17.5% vs 6.8%, p=0.048). L’analisi del polimorfismo rs8099917 ha fornito risultati sostanzialmente sovrapponibili. CONCLUSIONI: Nei pazienti HCV-positivi affetti da MC, il genotipo C/C rs12979860 del gene IL28B è caratterizzato biologicamente da una maggior frequenza di restrizione clonale nella risposta B-cellulare e clinicamente da un maggior rischio di nefropatia crioglobulinemica e neoplasie B-cellulari; è, inoltre, un fattore predittivo indipendente di risposta virologica sostenuta alla terapia antivirale. Nei pazienti con vasculite crioglobulinemica si è riscontrato una maggior prevalenza della variante T/T rs12979860 del gene IL28B. ABSTRACT INTRODUCTION: Hepatitis C virus (HCV) infection represents the leading cause of chronic liver damage evolving into cirrhosis and hepatocellular carcinoma (HCC). HCV chronic infection is also characterized by the emergence of several extrahepatic manifestations, the most common of which is represented by mixed cryoglobulinemia (MC). It is an immune complex-mediated vasculitis involving small vessels, characterized by an underlying B cell proliferation with potential evolution into B-cell non- Hodgkin lymphoma. The peculiar tropism of HCV for the lymphoid tissue has been clearly demonstrated. HCV persistence could represent a continuous stimulus to host immune system. It should lead to B cells oligo/monoclonal expansions with the emergence of a single dominant clone whose expression may be represented by monoclonal IgM RF production in MC. On the other hand, lymphocyte proliferation may be promoted by HCV direct infection and replication into B cells. On these bases the therapeutic approach of HCV-associated MC is currently based on antiviral therapy with or without B-cell depletion with rituximab. Among host factors influencing virological response to pegylated interferon- ibavirin-based antiviral therapeutic regimens, IL28B gene polymorphisms have been identified by genome wide association studies, as highly reliable predictors of the outcome after antiviral therapy. Aim of the study was to assess the possible association between IL28B single nucleotide polymorphism (SNPs) rs12979860/ rs8099917 and therapeutic response in patients with HCV-related MC. We also searched for their association with peculiar clinical manifestations of MC and the potential influence on the complete response (virological, molecular, and immunological) to the therapy. METHODS: Study cohort included 159 and 172 HCV infected patients with or without MC treated with pegylated interferon- -based antiviral therapy. We performed SNP rs12979860/ rs8099917 genotyping in all patients. In 106 (66.6%) patients with MC, the profile of circulating B cell clonalities was determined. RESULTS: The T/T IL-28B rs12979860 was more common in patients with MC than in those without (17% vs 8.1%, p = 0.02). C/C genotype was associated with a higher rate of complete response (52.6% vs 39.2%, p = 0.13). A higher frequency of expanded B cell clonalities in the circulation (84.2% vs 55.9%; p = 0.005), kidney involvement (21% vs 2.9%; p = 0.003), and B cell non-Hodgkin lymphoma (17.5% vs 6.8%; p = 0.048) were also observed. No additional results derived from the analysis of IL28B rs8099917 polymorphisms. CONCLUSIONS: In HCV-positive patients with MC, the IL-28B rs12979860 C/C genotype is distinguished biologically by a higher frequency of restriction of B cell response and clinically by a higher risk of cryoglobulinemic nephropathy and B cell malignancies, while acting as an independent predictor of a sustained virological response to antiviral therapy. In addition, we found that IL-28B rs12979860 T/T variant was more prevalent in patients with CV than in those without

Comparative analysis of cancer vaccine settings for the selection of an effective protocol in mice.

Background: Cancer vaccines are considered a promising therapeutic approach. However, their clinical results are not yet satisfactory. This may be due to the the difficulty of selection of an efficient tumor associated antigen (TAA) and immunization protocol. Indeed, the weak antigenicity of many TAA impairs the design of robust procedures, therefore a systematic analysis to identify the most efficient TAA is mandatory. Here, we performed a study to compare different gp100 vaccination strategies to identify the best strategy to provide a 100% protection against experimental melanoma in a reproducible manner. Methods: C57BL/6J mice were challenged subcutaneously with B16F10 melanoma cells, after vaccination with: a) mouse or human gp10025-33 peptide plus CpG adjuvant; b) mouse or human gp100 gene; c) mouse or human gp10025-33 peptide-pulsed dendritic cells (DC). Alternatively, a neutralizing anti-IL-10 monoclonal antibody (mAb) was subcutaneously administered at the site of tumor challenge to counteract regulatory cells. Finally, combinatorial treatment was performed associating human gp10025-33 peptide-pulsed DC vaccination with administration of the anti-IL-10 mAb. Results: Vaccination with human gp10025-33 peptide-pulsed DC was the most effective immunization protocol, although not achieving a full protection. Administration of the anti-IL-10 mAb showed also a remarkable protective effect, replicated in mice challenged with a different tumor, Anaplastic Large Cell Lymphoma. When immunization with gp10025-33 peptide-pulsed DC was associated with IL-10 counteraction, a 100% protective effect was consistently achieved. The analysis on the T-cell tumor infiltrates showed an increase of CD4+granzyme+ T-cells and a decreased number of CD4+CD25+Foxp3+ Treg elements from mice treated with either gp10025-33 peptide-pulsed DC vaccination or anti-IL-10 mAb administration. These data suggest that processes of intratumoral re-balance between effector and regulatory T cell subpopulations may play a critical protective role in immunotherapy protocols. Conclusions: Here we demonstrate that, in the setting of a cancer vaccine strategy, a comparative analysis of different personalized approaches may favour the unveiling of the most effective protocol. Moreover, our findings suggest that counteraction of IL-10 activity may be critical to revert the intratumoral environment promoting Treg polarization, thus increasing the effects of a vaccination against selected TAA.“Immunoterapia anti-tumorale: analisi d’efficacia dei principali protocolli tradizionali d’immunizzazione e validazione dell’efficacia terapeutica dell’inibizione dell’interleuchina 10 nel trattamento del melanom

Evaluation of the accuracy of a patient-specific instrumentation

Patient-specific instruments (PSI) has been introduced with the aim to reduce the overall costs of the implants, minimizing the size and number of instruments required, and also reducing surgery time. The aim of this study was to perform a review of the current literature, as well as to report about our personal experience, to assess reliability and accuracy of patient specific instrument system in total knee arthroplasty (TKA). A literature review was conducted of PSI system reviewing articles related to coronal alignment, clinical knee and function scores, cost, patient satisfaction and complications. Studies have reported incidences of coronal alignment ≥3° from neutral in TKAs performed with patient-specific cutting guides ranging from 6% to 31%. PSI seem not to be able to result in the same degree of accuracy as for the CAS system, while comparing well with standard manual technique with respect to component positioning and overall lower axis, in particular in the sagittal plane. In cases in which custom-made cutting jigs were used, we recommend performing an accurate control of the alignment before and after any cuts and in any further step of the procedure, in order to avoid possible outliers

In-out versus out-in technique for ACL reconstruction. a prospective clinical and radiological comparison

Background: Several studies have recently shown better restoration of normal knee kinematics and improvement of rotator knee stability after reconstruction with higher femoral tunnel obliquity. The aim of this study is to evaluate tunnel obliquity, length, and posterior wall blowout in single-bundle anterior cruciate ligament (ACL) reconstruction, comparing the transtibial (TT) technique and the out–in (OI) technique. Materials and methods: Forty consecutive patients operated on for ACL reconstruction with hamstrings were randomly divided into two groups: group A underwent a TT technique, while group B underwent an OI technique. At mean follow-up of 10 months, clinical results and obliquity, length, and posterior wall blowout of femoral tunnels in sagittal and coronal planes using computed tomography (CT) scan were assessed. Results: In sagittal plane, femoral tunnel obliquity was 38.6 ± 10.2° in group A and 36.6 ± 11.8° in group B (p = 0.63). In coronal plane, femoral tunnel obliquity was 57.8 ± 5.8° in group A and 35.8 ± 8.2° in group B (p = 0.009). Mean tunnel length was 40.3 ± 1.2 mm in group A and 32.9 ± 2.3 mm in group B (p = 0.01). No cases of posterior wall compromise were observed in any patient of either group. Clinical results were not significantly different between the two groups. Conclusions: The OI technique provides greater obliquity of the femoral tunnel in coronal plane, along with satisfactory length of the tunnel and lack of posterior wall compromise. Level of evidence: II, prospective study

REPLY TO THE LETTER TO THE EDITOR "ANDROGEN DEPRIVATION THERAPY AND RISK OF RHEUMATOID ARTHRITIS IN PATIENTS WITH LOCALIZED PROSTATE CANCER" by YANG et al.

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Nanophotonics for bacterial detection and antimicrobial susceptibility testing

Photonic biosensors are a major topic of research that continues to make exciting advances. Technology has now improved sufficiently for photonics to enter the realm of microbiology and to allow for the detection of individual bacteria. Here, we discuss the different nanophotonic modalities used in this context and highlight the opportunities they offer for studying bacteria. We critically review examples from the recent literature, starting with an overview of photonic devices for the detection of bacteria, followed by a specific analysis of photonic antimicrobial susceptibility tests. We show that the intrinsic advantage of matching the optical probed volume to that of a single, or a few, bacterial cell, affords improved sensitivity while providing additional insight into single-cell properties. We illustrate our argument by comparing traditional culture-based methods, which we term macroscopic, to microscopic free-space optics and nanoscopic guided-wave optics techniques. Particular attention is devoted to this last class by discussing structures such as photonic crystal cavities, plasmonic nanostructures and interferometric configurations. These structures and associated measurement modalities are assessed in terms of limit of detection, response time and ease of implementation. Existing challenges and issues yet to be addressed will be examined and critically discussed

L’uso di tecniche di similarità nell’editing di documenti fortemente strutturati

Lo scopo di questo progetto è quello di verificare la tesi secondo cui l’utilizzo di metodologie basate sul calcolo della similarità può migliorare l’editing di documenti strutturati attraverso attività di ricerca, recupero e confronto che forniscono supporto testuale a vari livelli di dettaglio. La ricerca di oggetti simili è un campo molto importante del Text Mining e, a tal proposito, sono state analizzate e implementate due tecniche che permettono di ricercare frammenti quali frasi, paragrafi e sezioni partendo da un testo in input e da una fissata metrica di similarità. Lo scopo è di progettare un sistema di supporto testuale che fornisca suggerimenti all’utente durante la fase di editing di un nuovo documento in modo da facilitare la scrittura e rendendo il documento conforme alle linee guida di una collezione di documenti iniziale. Nel corso della dissertazione verranno trattate tematiche come il concetto di similarità e i modi per quantificarla; verrà discusso come rappresentare vettorialmente il testo e, sulla base di ciò, verrà analizzato nel dettaglio gli algoritmi MRCTA e Minhash, un’istanza dello schema LSH (Locality Sensitive Hashing), che permette di stimare la similarità di Jaccard; verrà descritta SHE ossia l’ambiente implementato per consentire le fasi di confronto e testing delle tecniche considerate; infine, verranno analizzati i risultati della fase di test il cui scopo è quello di quantificare la soddisfazione dell’utente nell’utilizzo del sistema in modo da individuare, attraverso l’ausilio del questionario SUS, la tecnica più adatta

Pakkery: progettazione e sviluppo di un sistema di authoring basato su templating

La tesi ha lo scopo di presentare un sistema per il riuso controllato di contenuti su Web. Partendo dall’idea di transclusione di Nelson, abbiamo progettato e sviluppato Pakkery, un’applicazione web-based, che permette all’utente di creare documenti includendo all’interno di essi template. Oltre a velocizzare e facilitare l’editing, questo approccio favorisce la condivisione di documenti e l’aggiornamento diretto senza aver bisogno di modificare i documenti in cui queste risorse sono incluse. Come vedremo, in Pakkery un template è composto da due elementi: una parte fissa ed una variabile; quest’ultima permette di salvare le informazioni inserite dall’utente nel template a seguito di un aggiornamento della risorsa inclusa. Andremo ad analizzare lavori simili e precedenti a questo in modo da avere diversi punti di vista riguardo al concetto di riuso e di template; il secondo capitolo mira alla descrizione dell’architettura e dei componenti da cui è composta; il terzo analizza e mostra gli algoritmi utilizzati; l’ultimo capitolo mostra i risultati di un test preliminare effettuato su un numero ristretto di utenti