Microbiological characterization and effect of resin composites in cervical lesions

Non carious cervical lesions associated to muscle hyperfunctions are increasing. Microhybrid resin composites are used to restore cervical abfractions. The purpose of this study was to investigate if resin composites modify tooth plaque, inducing an increment of cariogenic microflora and evaluate their effect, in vivo and in vitro, against S. mutans. Eight abfractions were restored with two microhybrid resin composites (Venus, Heraeus-Kulzer® and Esthet-X, Dentsply®), after gnatological therapy, in three patients with muscle hyperfunctions. For each abfraction three samples of plaque were taken from the cervical perimeter: before the restoration, one week and three months after restoration. The samples were evaluated both by traditional microbiological methods and by Polymerase Chain Reaction (PCR). In vitro, disk-shaped specimens of the two composites were prepared to estimate the effects against pre-cultured S. mutans, after incubation at 37°C for 24h and assessed by a turbidimetric technique. In vivo no differences were found in plaque growth, for all samples, before and after restoration with both composites; in vitro, instead, a significant reduction of S. mutans growth was found between specimens of two composites (Mann-Whitney U-test p>0,06). In this study a relevant consideration was elicited: composite materials, in vivo, do not modify plaque composition of non carious cervical lesions to a potential cariogenic plaque

DEGASSAMENTO DI RADON E MODIFICHE DEL DNA DELLA POPOLAZIONE SICILIANA: POSSIBILI RELAZIONI CON IL TERREMOTO DEL 1908

I risultati delle indagini condotte su base geochimica e genetica nell’area delle Stretto di Messina si incrociano proponendo uno scenario assolutamente nuovo su ciò che un terremoto può indurre anche come effetti sulla salute umana, con modifiche di tipo genetico. Le ricerche svolte nell’ambito del progetto INGV-Dipartimento Protezione Civile su “Valutazione del potenziale sismogenetico e probabilità dei forti terremoti in Italia”, hanno mostrato che nell’area dello Stretto di Messina persistono anomalie di degassamento dai suoli di CO2, metano e radon, diversi ordini di grandezza al di sopra dei valori normali, per esempio con concentrazioni dinamiche di Rn fino oltre 80.000 Bq/m3 La valutazione di un degassamento naturale di tale intensità in situazione di quiete sismica, ha assunto significato anche nella valutazione degli effetti del Rn sulla salute umana, che, sebbene conosciuti da tempo come seconda causa di tumore al polmone dopo il fumo, si mostrano sorprendentemente come i possibili protagonisti delle mutazioni genetiche riscontrate nella popolazione vivente intorno allo Stretto Radon e DNA La presenza di antigeni indicano di modifiche del DNA, ed uno in particolare denominato DR11 è presente con la frequenza più alta intorno a Messina e a Reggio (54% della popolazione) per poi diminuire verso Caltanissetta e Vibo Valentia (44%) con un minimo a Trapani (38%; figura 1). Una tale modifica del DNA è in contrasto con la Legge di Hardy –Weimberg, secondo la quale in una popolazione in cui gli accoppiamenti avvengono a caso, e in assenza di pressioni evolutive e di forti migrazioni dovrebbe essere mantenuta una generale omogeneità. Questo tipo di distribuzione, molto eterogenea, ma non irregolare e disordinata, non può essere attribuita a fluttuazioni casuali o a deriva genetica, e neanche alle invasioni che nel passato queste regioni hanno subito da parte di popoli stranieri.L’ordinata e graduale riduzione di frequenza del DR11, funzione dalla distanza dallo stretto, unitamente alle valutazioni basate sulla riduzione nel tempo della frequenza del DR11, portano a concentrare l’attenzione su un evento con epicentro nello stretto accaduto circa un secolo fa, identificabile come il terremoto del 1908. Tenuto conto che non ci sono effetti diretti di un sisma in grado di interagire con la saluteumana (se non ferite, paura) si ipotizza che non sia direttamente il terremoto come tale la causa scatenante della mutazione genetica osservata ma qualcosa che ad un forte evento sismico possa essere legata. Fluidi e terremoti I risultati ottenuti in varie aree sismiche italiane (p.e. Umbria, Friuli, Appennino Tosco Emiliano) hanno mostrato che i fluidi cambiamo composizione e intensità del rilascio durante l’evolversi della sismogenesi. Le modifiche si registrano prima, durante e dopo gli eventi sismici mostrando che oltre alla fratturazione anche le deformazioni della crosta provocano modificazioni nei fluidi circolanti. Una aumentata emissione di radon durante il processo che ha portato al terremoto del 1908 è assolutamente ipotizzabile in accordo con quanto osservato durante la crisi sismica dell’Umbria (1997-98) e con le misure effettuate nell’area dello Stretto (Figura 2). Una contaminazione da gas radon dovuta alla sismogenesi, che si è protratta per tempi probabilmente lunghi, prima durante e dopo il catastrofico evento, può essere la causa delle mutazioni genetiche osservate, avendo indotto gli organismi a reagire per tutelarsi rispetto ad una pressione esterna pericolosa.Un simile scenario, che vede i processi sismogenetici strettamente legati ai fluidi anche con attivi processi di degassamento, richiede ulteriori indagini che oltre a contribuire alla riduzione del rischio sismico permettono la conoscenza di quegli aspetti legati alle interazioni uomo-ambiente su medio-lungo termine anche come possibili responsabili di cambiamenti profondi come quelli identificati del DNA dei siciliani

Nosocomial outbreak of the pandemic Influenza A (H1N1) 2009 in critical hematologic patients during seasonal influenza 2010-2011: detection of oseltamivir resistant variant viruses

BACKGROUND: The pandemic influenza A (H1N1) 2009 (H1N1pdm09) virus infection caused illness and death among people worldwide, particularly in hematologic/oncologic patients because influenza infected individuals can shed virus for prolonged periods, thus increasing the chances for the development of drug-resistant strains such as oseltamivir-resistant (OST-r) variant. METHODS: The aim of our study was to retrospectively evaluate the clinical importance of OST-r variant in circulating strains of the pandemic H1N1pdm09 virus. By means of RT-PCR and Sanger sequencing we analysed the presence of OST-r variant in 76 H1N1pdm09 laboratory-confirmed cases, hospitalized at the hematologic/oncologic ward at Spedali Civili of Brescia –Italy. RESULTS: Out of 76 hospitalized hematologic/oncologic patients, 23 patients (30.2%) were infected by H1N1pdm09 virus. Further investigation revealed that 3 patients were positive for the OST-r variant carrying the H275Y mutation. All the 23 infected patients were immuno-compromised, and were under treatment or had been treated previously with oseltamivir. Three patients died (13%) after admission to intensive care unit and only one of them developed H275Y mutation. CONCLUSIONS: Our retrospective observational study shows that pandemic influenza A (H1N1) 2009 virus can cause significant morbidity and even mortality in hematologic/oncologic patients and confirms the high rate of nosocomial transmission of pandemic H1N1pdm09 virus in these critical subjects. Indeed, the reduction in host defences in these hospitalized patients favoured the prolonged use of antiviral therapy and permitted the development of OST-r strain. Strategies as diagnostic vigilance, early isolation of patients and seasonal influenza A(H1N1) vaccination may prevent transmission of influenza in high risk individuals

AB012. Transcriptional and chromatin profiling reveals the molecular architecture and druggable vulnerabilities of thymic epithelial tumors (TETs)

Thymic epithelial tumors (TETs) have been profiled to the present moment mainly through several analyses of FFPE samples. Despite the leap forward brought by the TCGA, several questions remain still unsolved. Among these, TETs are characterized by a strong component of immune infiltrate which makes the transcriptomic analyses conducted so far scarcely interpretable to profile stromal subpopulations constitutive of the tumor. Furthermore, rarely correspondent healthy tissue is available due to the lipomatous atrophy of aged thymi. Therefore, the recent report of (I) isolation, (II) propagation (III) and characterization of human thymic epithelial cells (TECs) and their capacity to reconstitute the functional organ ex vivo and in vivo, represents a novel approach to study the biology of both healthy and neoplastic thymi. Human thymic biopsies (both healthy and neoplastic) were digested and plated on a lethally irradiated murine feeder layer. Both RNA-Seq and CUTANDTAG were performed on cultivated TECs at different passages. Cultured TECs were injected with human thymic interstitial cells into rat decellularized scaffolds and cultivated for 10–12 days. sc-RNA Seq is currently being performed on both healthy and neoplastic thymic mini-organs and their correspondent primary tissues. Here show that we successfully cultivated a cohort of 21 clonogenic TECs in vitro including adult neoplastic TECs, their non-tumoral counterpart and pediatric TECs. We show that at the transcriptome level each class of TECs clusters independently and that neoplastic TECs belong to the same cloud independently from thymoma histotype. Around 1,400 differentially expressed genes (DEGs) can be found when comparing adult neoplastic and non-neoplastic counterpart, among which around 70 are transcription factors. Importantly, we prove for the first time that clonogenic TECs derived from TETs can repopulate a decellularized rat scaffold and recreate a 3D architecture mimicking the primary tumor. This work demonstrates that this culture system allows the expansion of clonogenic TECs from both tumor samples and their non-tumoral counterpart. Those cells, when transplanted into decellularized thymi, reproduce the architecture of the primary tissue, showing that TETs contain progenitor/stem epithelial cells. We are currently characterizing TECs at the transcriptomic and epigenomic level with aim of identifying new druggable targets prior to clinical trials

Reconstitution of a functional human thymus by postnatal stromal progenitor cells and natural whole-organ scaffolds.

The thymus is a primary lymphoid organ, essential for T cell maturation and selection. There has been long-standing interest in processes underpinning thymus generation and the potential to manipulate it clinically, because alterations of thymus development or function can result in severe immunodeficiency and autoimmunity. Here, we identify epithelial-mesenchymal hybrid cells, capable of long-term expansion in vitro, and able to reconstitute an anatomic phenocopy of the native thymus, when combined with thymic interstitial cells and a natural decellularised extracellular matrix (ECM) obtained by whole thymus perfusion. This anatomical human thymus reconstruction is functional, as judged by its capacity to support mature T cell development in vivo after transplantation into humanised immunodeficient mice. These findings establish a basis for dissecting the cellular and molecular crosstalk between stroma, ECM and thymocytes, and offer practical prospects for treating congenital and acquired immunological diseases

Relationship between pp65 antigenemia levels and real-time quantitative DNA PCR for Human Cytomegalovirus (HCMV) management in immunocompromised patients

<p>Abstract</p> <p>Background</p> <p>Quantitative real-time PCR assays, which are more rapid and practical than pp65 antigenemia determination, are progressively becoming the preferred method for monitoring Human Cytomegalovirus (HCMV) reactivation. However, the relationship between HCMV DNA and antigenemia levels is still under investigation. The aim of this study was to analyse the relationship between HCMV DNA and pp65 antigenemia levels in order to identify clinically useful threshold values for the management of patients.</p> <p>Methods</p> <p>475 consecutive samples from 156 immunosuppressed patients were tested for HCMV by pp65 antigenemia and Real-time PCR assay.</p> <p>Results </p> <p>136 out of 475 consecutive samples derived from 48 patients showed evidence of HCMV infection. HCMV DNA was detected in 106 samples, pp65 antigen in 3, and both markers in 27. pp65 antigen detection was associated with higher HCMV DNA levels. The cut-off HCMV DNA level that best predicted pp65 antigenemia in this series of samples was 11,500 copies/ml, but different threshold levels could be observed for specific groups of patients. HCMV disease was observed in 5 out of 48 patients with active HCMV infection. The presence of clinical symptoms was associated with positive pp65 and with higher antigenemia levels. Higher HCMV DNA load at the onset of viral replication was correlated to the development of clinical symptoms.</p> <p>Conclusion</p> <p>Both pp65 antigenemia and HCMV DNA load can be useful for the prospective monitoring of immunocompromised subjects. Specific cut-off levels capable of triggering preemptive antiviral treatment should be determined in accordance to the type of test used and the characteristics of patients and prospectively validated.</p

Lopinavir/Ritonavir and Darunavir/Cobicistat in Hospitalized COVID-19 Patients: Findings From the Multicenter Italian CORIST Study

Background: Protease inhibitors have been considered as possible therapeutic agents for COVID-19 patients. Objectives: To describe the association between lopinavir/ritonavir (LPV/r) or darunavir/cobicistat (DRV/c) use and in-hospital mortality in COVID-19 patients. Study Design: Multicenter observational study of COVID-19 patients admitted in 33 Italian hospitals. Medications, preexisting conditions, clinical measures, and outcomes were extracted from medical records. Patients were retrospectively divided in three groups, according to use of LPV/r, DRV/c or none of them. Primary outcome in a time-to event analysis was death. We used Cox proportional-hazards models with inverse probability of treatment weighting by multinomial propensity scores. Results: Out of 3,451 patients, 33.3% LPV/r and 13.9% received DRV/c. Patients receiving LPV/r or DRV/c were more likely younger, men, had higher C-reactive protein levels while less likely had hypertension, cardiovascular, pulmonary or kidney disease. After adjustment for propensity scores, LPV/r use was not associated with mortality (HR = 0.94, 95% CI 0.78 to 1.13), whereas treatment with DRV/c was associated with a higher death risk (HR = 1.89, 1.53 to 2.34, E-value = 2.43). This increased risk was more marked in women, in elderly, in patients with higher severity of COVID-19 and in patients receiving other COVID-19 drugs. Conclusions: In a large cohort of Italian patients hospitalized for COVID-19 in a real-life setting, the use of LPV/r treatment did not change death rate, while DRV/c was associated with increased mortality. Within the limits of an observational study, these data do not support the use of LPV/r or DRV/c in COVID-19 patients