Background Glioblastoma (GBM) stemlike cells (GSCs) are thought to be responsible for the maintenance and aggressiveness of GBM, the most common primary brain tumor in adults. This study aims at elucidating the involvement of deregulations within the imprinted delta-like homolog 1 gene‒type III iodothyronine deiodinase gene (DLK-DIO3) region on chromosome 14q32 in GBM pathogenesis. Methods Real-time PCR analyses were performed on GSCs and GBM tissues. Methylation analyses, gene expression, and reverse-phase protein array profiles were used to investigate the tumor suppressor function of the maternally expressed 3 gene (MEG3). Results Loss of expression of genes and noncoding RNAs within the DLK1-DIO3 region was observed in GSCs and GBM tissues compared with normal brain. This downregulation is mainly mediated by epigenetic silencing. Kaplan–Meier analysis indicated that low expression of MEG3 and MEG8 long noncoding (lnc)RNAs significantly correlated with short survival in GBM patients. MEG3 restoration impairs tumorigenic abilities of GSCs in vitro by inhibiting cell growth, migration, and colony formation and decreases in vivo tumor growth, reducing infiltrative growth. These effects were associated with modulation of genes involved in cell adhesion and epithelial-to-mesenchymal transition (EMT). Conclusion In GBM, MEG3 acts as a tumor suppressor mainly regulating cell adhesion, EMT, and cell proliferation, thus providing a potential candidate for novel GBM therapies.

Background: Preterm infants often receive blood transfusions early in life. In this setting, umbilical cord blood (UCB) might be safer than adult blood (A) with respect to infectious and immunologic threats. Objectives: To evaluate, as a first objective, the feasibility of fulfilling transfusion needs of preterm infants with allogeneic UCB red blood cell (RBC) concentrates and, as a secondary objective, to assess the safety of allogeneic cord blood transfusions. Methods: At the Neonatal Intensive Care Unit and the UNICATT Cord Blood Bank of ‘A. Gemelli' Hospital in Rome, a prospective study was carried out over a 1-year period, enrolling newborns with gestational age ≤30 weeks and/or birth weight ≤1,500 g requiring RBC transfusions within the first 28 days of life. At first transfusion, patients were assigned to receive UCB-RBCs or A-RBCs depending on the availability of ABO-Rh(D)-matched UCB-RBC units. The same regimen (UCB-RBC or A-RBC units) was thereafter maintained, unless ABO-Rh(D)-matched UCB-RBC units were not available. Results: Overall, 23 UCB-RBC units were transfused to 9 patients; the requests for UCB-RBC units were met in 45% of patients at the first transfusion and in 78% at the subsequent transfusions. At a median follow-up of 57 days (range 6-219), no acute or delayed transfusion-related adverse events occurred. Hematocrit gain after transfusion and time intervals between transfusions were similar in the UCB-RBC and A-RBC group, as well. Conclusions: Transfusing allogeneic UCB-RBC units in preterm infants appears a feasible and safe approach, although the transfusion needs of our study population were not completely covered. More data are necessary to validate this novel transfusion practice. © 2014 S. Karger AG, Basel

BackgroundWe retrospectively investigated the incidence and risk factors for transfusion-related acute lung injury (TRALI) among patients transfused for post-partum hemorrhage (PPH).MethodsWe identified a series of 71 consecutive patients with PPH requiring the urgent transfusion of three or more red blood cell (RBC) units, with or without transfusion of fresh frozen plasma (FFP) and/or platelets (PLT). Clinical records were then retrieved and examined for respiratory distress events. According to the 2004 consensus definition, cases of new-onset hypoxemia, within 6 hours after transfusion, with bilateral pulmonary changes, in the absence of cardiogenic pulmonary edema were identified as TRALI. If an alternative risk factor for acute lung injury was present, possible TRALI was diagnosed.ResultsThirteen cases of TRALI and 1 case of possible TRALI were identified (overall incidence 19.7%). At univariate analysis, patients with TRALI received higher number of RBC, PLT and FFP units and had a longer postpartum hospitalization. Among the diseases occurring in pregnancy- and various pre-existing comorbidities, only gestational hypertension and pre-eclampsia, significantly increased the risk to develop TRALI (p = 0.006). At multivariate analysis including both transfusion- and patient-related risk factors, pregnancy-related, hypertensive disorders were confirmed to be the only predictors for TRALI, with an odds ratio of 27.7 ( 95% CI 1.27–604.3, p=0.034).ConclusionsPatients suffering from PPH represent a high-risk population for TRALI. The patients with gestational hypertension and pre-eclampsia, not receiving anti-hypertensive therapy, have the highest risk. Therefore, a careful monitoring of these patients after transfusions is recommended.

Background:Spinal muscular atrophy (SMA) is a neuromuscular disorder characterized by the degeneration of the second motor-neuron. The phenotype ranges from very severe to very mild forms. All patients have the homozygous loss of the SMN1 gene and a variable number of SMN2 (generally two-to-four copies), inversely related with the severity. The amazing results of the available treatments have made compelling the need of prognostic biomarkers to predict the progression trajectories of patients. Beside the SMN2 products, few other biomarkers have been evaluated so far, including some miRs.Methods:We performed whole miRNome analysis of muscle samples of patients and controls (14 biopsies and 9 cultures). The levels of muscle differentially expressed miRs were evaluated in serum samples (51 patients and 37 controls) and integrated with SMN2 copies, SMN2-full length transcript levels in blood and age (SMA-score). Results:Over 100 miRs were differentially expressed in SMA muscle; three of them (HSA-miR-181a-5p, -324-5p, -451a; SMA-miRs) were significantly up-regulated in serum of patients. The severity predicted by the SMA-score was related with that of the clinical classification at a correlation coefficient of 0.87 (p<10-5).Conclusions:miRNome analyses suggest the primary involvement of skeletal muscle in SMA pathogenesis; the SMA-miRs are likely actively released in the blood flow, even if their function and target cells require to be elucidated. The accuracy of the SMA-score needs to be verified in replicative studies: if confirmed, its use could be crucial for the routine prognostic assessment, also in pre-symptomatic patients. Funding:Telethon Italia (grant # GGP12116).

BackgroundSpinal muscular atrophy (SMA) is due to the homozygous absence ofSMN1in around 97% of patients, independent of the severity (classically ranked into types I–III). The high genetic homogeneity, coupled with the excellent results of presymptomatic treatments of patients with each of the three disease-modifying therapies available, makes SMA one of the golden candidates to genetic newborn screening (NBS) (SMA-NBS). The implementation of SMA in NBS national programmes occurring in some countries is an arising new issue that the scientific community has to address. We report here the results of the first Italian SMA-NBS project and provide some proposals for updating the current molecular diagnostic scenario.MethodsThe screening test was performed by an in-house-developed qPCR assay, amplifyingSMN1andSMN2. Molecular prognosis was assessed on fresh blood samples.ResultsWe found 15 patients/90885 newborns (incidence 1:6059) having the followingSMN2genotypes: 1 (one patient), 2 (eight patients), 2+c.859G>C variant (one patient), 3 (three patients), 4 (one patient) or 6 copies (one patient). Six patients (40%) showed signs suggestive of SMA at birth. We also discuss some unusual cases we found.ConclusionThe molecular diagnosis of SMA needs to adapt to the new era of the disease with specific guidelines and standard operating procedures. In detail, SMA diagnosis should be felt as a true medical urgency due to therapeutic implications;SMN2copy assessment needs to be standardised; commercially available tests need to be improved for higherSMN2copies determination; and theSMN2splicing-modifier variants should be routinely tested in SMA-NBS.

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Infants with very low birth weight (VLBW) frequently need transfusions of red blood cells (RBCs) during the first weeks of life. However, adult blood transfusions are acknowledged risk factors for several complications, including Retinopathy of Prematurity (ROP), cytomegalovirus infection and necrotizing enterocolitis. For ROP, for example, it is thought that adult hemoglobin (HbA), with lower affinity for oxygen than foetal HbF, may induce an oxidative damage (Romagnoli C. Early Hum Dev 2009; 85, 10 Suppl:S79-S82). Previous studies showed that autologous cord blood (CB) could serve as source of RBCs for transfuse neonates; nevertheless, his clinical use is still limited, expecially because of the small volumes achieved after processing of the UCB unit. In a previous study we demonstrated that CB derived buffy coat–depleted RBC units obtained through automated separation (Compomat G4®, Fresenius HemoCare, Germany) and stored in SAG-Mannitol solution represent a suitable product for homologous transfusion of neonates. Actually, CB RBC units show hemoglobin content and hematocrit (Htc) values similar to adult RBC units stored for comparable periods, whereas the lactate concentrations are lower and the pH values are higher (Bianchi et al. ASH Annual Meeting Abstracts 2012, 120:275). We are now assessing the feasibility of covering the transfusion needs of VLBW neonates using allogeneic CB packed RBC units collected at our Cord Blood Bank. This practice has never been used before and we show here for the first time our experience on allogeneic CB derived RBC transfusion. VLBW neonates admitted to the Neonatal Intensive Care Unit needing RBC transfusions in the first 28 days of life receive adult (A group) or CB (CB group) RBC units, on the basis of the availability of an AB0-Rh compatible CB RBC unit. The arm assignment drives the choice of the RBC products (adult versus CB) in case of subsequent transfusions. All patients receive a fixed dose of 20 ml/kg RBC. Htc values are acquired before and after transfusion (ΔHtc). CB RBC units are processed and stored as previously reported (Bianchi et al. ASH Annual Meeting Abstracts 2012, 120:275). After matching tests, CB and adult units are irradiated and filtered; the Purecell RN Neonatal filters (PALL Medical, UK) are used for CB units. So far, 9 patients entered the study. Five patients are in the A group and 4 in the CB group, with similar gestational age (mean 27 + 1 weeks in group A and 27 + 3 weeks in group CB, respectively), gender (male/female ratio 3/1 in A group and 4/1 in CB group, respectively) birth weight (mean weight 915 + 225 gr in A group and 918 + 389 gr in B group, respectively) and Htc values at birth (57.2 + 8.4% in A group and 55.5 + 5.2% in CB group, respectively). On the whole, 21 RBC units were transfused, 7 in the A group and 14 in the CB group. The mean storage time was 5 + 4 days for adult RBC units and 9 + 3 days for CB units. The Htc values of patients at the time of transfusion were similar in the two groups (32,2 + 4% in A group and 30.64 + 4% in CB group, respectively, p=0.287). The ΔHtc was similar in A and CB groups, with a mean increase of 15.1 + 5 % in A group and 13.3 + 5% in the CB group, respectively, p = 0,426). No transfusion related adverse event occurred in both arms. CB can be safely administrated to preterm infants: as compared to adult, cord blood is functionally more appropriate and is safer for infectious and immunological complications. Given the wide availability of discharged units at public cord blood banks, CB derived RBC transfusion can constitute a valid therapeutic option for VLBW neonates. The study was supported by a grant from Genitin (Associazione dei Genitori per la Terapia Intensiva Neonatale). Disclosures: No relevant conflicts of interest to declare.