Heteroepitaxy of deposited amorphous layer by pulsed electron-beam irradiation

We demonstrate that a single short pulse of electron irradiation of appropriate energy is capable of recrystallizing epitaxially an amorphous Ge layer deposited on either or Si single-crystal substrate. The primary defects observed in the case were dislocations, whereas stacking faults were observed in samples

Autologus peripheral stem cell transplantation in a patient with diffuse systemic sclerosis: our experience

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Clinical presentations leading to arrhythmogenic left ventricular cardiomyopathy

Objectives To describe a cohort of patients with arrhythmogenic left ventricular cardiomyopathy (ALVC), focusing on the spectrum of the clinical presentations. Methods Patients were retrospectively evaluated between January 2012 and June 2020. Diagnosis was based on (1) ≥3 contiguous segments with subepicardial/midwall late gadolinium enhancement in the left ventricle (LV) at cardiac magnetic resonance plus a likely pathogenic/pathogenic arrhythmogenic cardiomyopathy (AC) associated genetic mutation and/or familial history of AC and/or red flags for ALVC (ie, negative T waves in V4-6/aVL, low voltages in limb leads, right bundle branch block like ventricular tachycardia) or (2) pathology examination of explanted hearts or autoptic cases suffering sudden cardiac death (SCD). Significant right ventricular involvement was an exclusion criterion. Results Fifty-two patients (63% males, age 45 years (31-53)) composed the study cohort. Twenty-one (41%) had normal echocardiogram, 13 (25%) a hypokinetic non-dilated cardiomyopathy (HNDC) and 17 (33%) a dilated cardiomyopathy (DCM). Of 47 tested patients, 29 (62%) were carriers of a pathogenic/likely pathogenic DNA variant. Clinical contexts leading to diagnosis were SCD in 3 (6%), ventricular arrhythmias in 15 (29%), chest pain in 8 (15%), heart failure in 6 (12%) and familial screening in 20 (38%). Thirty patients (57%) had previously received a diagnosis other than ALVC with a diagnostic delay of 6 years (IQR 1-7). Conclusions ALVC is hidden in different clinical scenarios with a phenotypic spectrum ranging from normal LV to HNDC and DCM. Ventricular arrhythmias, chest pain, heart failure and SCD are the main clinical presentations, being familial screening essential for the affected relatives' identification

Frequency and duration of SARS-CoV-2 shedding in oral fluid samples assessed by a modified commercial rapid molecular assay

Background: RT-PCR on nasopharyngeal (NPS)/oropharyngeal swabs is the gold standard for diagnosis of SARS-CoV-2 infection and viral load monitoring. Oral fluid (OF) is an alternate clinical sample, easy and safer to collect and could be useful for COVID-19 diagnosis, monitoring viral load and shedding. Methods: Optimal assay conditions and analytical sensitivity were established for the commercial Simplexa™ COVID-19 Direct assay adapted to OF matrix. The assay was used to test 337 OF and NPS specimens collected in parallel from 164 hospitalized patients; 50 bronchoalveolar lavage (BAL) specimens from a subgroup of severe COVID-19 cases were also analysed. Results: Using Simplexa™ COVID-19 Direct on OF matrix, 100% analytical detection down to 1 TCID50/mL (corresponding to 4 × 103 copies (cp)/mL) was observed. No crossreaction with other viruses transmitted through the respiratory toute was observed. Parallel testing of 337 OF and NPS samples showed highly concordant results (κ = 0.831; 95 % CI = 0.771–0.891), and high correlation of Ct values (r = 0.921; p < 0.0001). High concordance and elevated correlation was observed also between OF and BAL. Prolonged viral RNA shedding was observed up to 100 days from symptoms onset (DSO), with 32% and 29% positivity observed in OF and NPS samples, respectively, collected between 60 and 100 DSO. Conclusions: Simplexa™ COVID-19 Direct assays on OF have high sensitivity and specificity to detect SARS-CoV-2 RNA and provide an alternative to NPS for diagnosis and monitoring SARS-CoV-2 shedding

Le confessioni religiose tra libertà di vivere nella realtà dell'ordinamento statale e potere di creare norme giuridiche all'interno dello Stato. Il caso della Chiesa di Scientology.

L'A. si pone nella prospettiva di delineare il possibile significato della compresenza delle due distinte previsioni, l’una nell’art. 8, l’altra nell’art. 19 della Costituzione, a garanzia del fenomeno associativo religioso. La ricerca è stata redatta, fra l’altro, in considerazione della circostanza che, ad una prima sommaria lettura della Carta Costituzionale, il lettore potrebbe essere indotto a pensare che il fenomeno religioso, ed in particolare quello che si svolge in forma associata, goda di una maggiore preferenza, rispetto a quello che si esprime in forma individuale, per essere più volte disciplinato in Costituzione. Problema, questo, il quale non sembra abbia sinora ricevuto adeguato rilievo dalla dottrina ecclesiasticista che si è meritevolmente impegnata sul tema. Si è posto al riguardo l’interrogativo che nasce dalla presenza di due distinte previsioni costituzionali a garanzia del fenomeno associativo religioso: la prima contenuta nell’art. 8 il quale - com’è noto - regola con novità di linguaggio la posizione dei culti diversi dalla religione cattolica, garantisce alle confessioni religiose l’eguale libertà davanti alla legge, il diritto di organizzarsi secondo i propri statuti ed il potere di concludere intese con gli organi dello Stato; la seconda nell’art. 19 che riconosce la libertà di professare liberamente la propria fede religiosa in qualsiasi forma “individuale o associata”. In ordine ai numerosi problemi che la previsione costituzionale dell’art. 8 solleva, si è riflettuto soprattutto sul dubbio se la fattispecie in essa contemplata sia sostanzialmente ripetitiva e si risolva in un inutile duplicato di quanto disposto dall’art. 19 per la parte che riguarda tale aspetto, oppure se, con ciascuna di tali disposizioni, il legislatore costituzionale abbia voluto conferire uno specifico rilievo a due distinte sfere dell’esperienza giuridica con caratteristiche proprie, per struttura e sistemi di garanzie. L’analisi intrapresa ha interessato anche la ricostruzione della definizione giuridica di confessione religiosa, nonché l’accertamento del carattere confessionale della Chiesa di Scientology alla luce delle sentenze della giurisprudenza costituzionale e di legittimità

Simple, rapid and accurate molecular diagnosis of acute promyelocytic leukemia by loop mediated amplification technology

The diagnostic work-up of acute promyelocytic leukemia (APL) includes the cytogenetic demonstration of the t(15;17) translocation and/or the PML-RARA chimeric transcript by RQ-PCR or RT-PCR. This latter assays provide suitable results in 3-6 hours. We describe here two new, rapid and specific assays that detect PML-RARA transcripts, based on the RT-QLAMP (Reverse Transcription-Quenching Loop-mediated Isothermal Amplification) technology in which RNA retrotranscription and cDNA amplification are carried out in a single tube with one enzyme at one temperature, in fluorescence and real time format. A single tube triplex assay detects bcr1 and bcr3 PML-RARA transcripts along with GUS housekeeping gene. A single tube duplex assay detects bcr2 and GUSB. In 73 APL cases, these assays detected in 16 minutes bcr1, bcr2 and bcr3 transcripts. All 81 non-APL samples were negative by RT-QLAMP for chimeric transcripts whereas GUSB was detectable. In 11 APL patients in which RT-PCR yielded equivocal breakpoint type results, RT-QLAMP assays unequivocally and accurately defined the breakpoint type (as confirmed by sequencing). Furthermore, RT-QLAMP could amplify two bcr2 transcripts with particularly extended PML exon 6 deletions not amplified by RQ-PCR. RT-QLAMP reproducible sensitivity is 10(-3) for bcr1 and bcr3 and 10(-)2 for bcr2 thus making this assay particularly attractive at diagnosis and leaving RQ-PCR for the molecular monitoring of minimal residual disease during the follow up. In conclusion, PML-RARA RT-QLAMP compared to RT-PCR or RQ-PCR is a valid improvement to perform rapid, simple and accurate molecular diagnosis of APL

THE IMPROVED DIASORIN Q-LAMP ASSAY FOR THE ACCURATE AND ULTRA-FAST DETECTION OF COMMON AND RARE ISOFORMS OF THE BCR-ABL1 TRANSLOCATION

The molecular detection of BCR-ABL1 transcripts by RT-PCR is mandatory for the diagnosis of Ph+ Leukemias at onset. Recently a faster and reliable assay based on the Q-LAMP technology developed by DiaSorin has entered in laboratory routine. This assay detects in one hour BCR-ABL1 p190 (e1a2) and p210 (e13a2, e14a2) isoforms. In this study, we evaluated the new improved Q-LAMP formulation designed to detect also less frequent isoforms of the BCR-ABL1 transcripts p190 and p210 (e1a3, e13a3, e14a3). In addition, clinical studies demonstrated that the assay is capable of detecting also the rare isoform p230 (e19a2, e19a3). Methods: The new Q-LAMP technology consists in a multiplex assay for the differential detection of p190 and p210 transcripts and the amplification of the GUSB endogenous RNA. The assay has been tested on 185 clinical samples including 95 p210 positive (57 e13a2 and 28 e14a2, 8 e13a3 and 2 e14a3), 38 p190 positive (33 e1a2 and 5 e1a3) and 50 BCR-ABL1 negative samples. Additional 2 p230 rare isoforms were also included in this study. All samples were previously tested by RT-PCR, considered as the reference method. Results: The new BCR-ABL Q-LAMP assay showed 100% concordance with the RT-PCR, with an expected delayed amplification time for rare isoforms respect to the common ones. The average amplification time of p210 common isoforms were 22,24 and 25,03 min compared to the p210 and p190 rare isoforms that showed 26,54 and 36,84 min, respectively. The 2 p230 (e19a2) rare isoforms were also tested and resulted valid although, due to the very long transcript, they showed a very high average amplification time (50 and 48 min). Moreover, we observed an interesting discrimination between the e13a2 and the e14a2 isoforms in terms of amplification times (20,21 versus 26,36 min) likely associated to the different length of the two transcripts, with low coefficients of variability (0,15 and 0,11 respectively). Conclusions: The enhanced BCR-ABL Q-LAMP assay well proved to detect both common and uncommon isoforms of the BCR-ABL1 translocation. This improved performances, combined with the speed and the close tube format, allow laboratories to optimize their workflow and represent a reliable solution for molecular diagnosis of Philadelphia Positive Leukemias

Association of kidney disease measures with risk of renal function worsening in patients with type 1 diabetes

Background: Albuminuria has been classically considered a marker of kidney damage progression in diabetic patients and it is routinely assessed to monitor kidney function. However, the role of a mild GFR reduction on the development of stage 653 CKD has been less explored in type 1 diabetes mellitus (T1DM) patients. Aim of the present study was to evaluate the prognostic role of kidney disease measures, namely albuminuria and reduced GFR, on the development of stage 653 CKD in a large cohort of patients affected by T1DM. Methods: A total of 4284 patients affected by T1DM followed-up at 76 diabetes centers participating to the Italian Association of Clinical Diabetologists (Associazione Medici Diabetologi, AMD) initiative constitutes the study population. Urinary albumin excretion (ACR) and estimated GFR (eGFR) were retrieved and analyzed. The incidence of stage 653 CKD (eGFR < 60 mL/min/1.73 m2) or eGFR reduction > 30% from baseline was evaluated. Results: The mean estimated GFR was 98 \ub1 17 mL/min/1.73m2 and the proportion of patients with albuminuria was 15.3% (n = 654) at baseline. About 8% (n = 337) of patients developed one of the two renal endpoints during the 4-year follow-up period. Age, albuminuria (micro or macro) and baseline eGFR < 90 ml/min/m2 were independent risk factors for stage 653 CKD and renal function worsening. When compared to patients with eGFR > 90 ml/min/1.73m2 and normoalbuminuria, those with albuminuria at baseline had a 1.69 greater risk of reaching stage 3 CKD, while patients with mild eGFR reduction (i.e. eGFR between 90 and 60 mL/min/1.73 m2) show a 3.81 greater risk that rose to 8.24 for those patients with albuminuria and mild eGFR reduction at baseline. Conclusions: Albuminuria and eGFR reduction represent independent risk factors for incident stage 653 CKD in T1DM patients. The simultaneous occurrence of reduced eGFR and albuminuria have a synergistic effect on renal function worsening