Search

Background Gram-negative bacteremia (GNB) is a major cause of illness and death after hematopoietic stem cell transplantation (HSCT), and updated epidemiological investigation is advisable. Methods We prospectively evaluated the epidemiology of pre-engraftment GNB in 1118 allogeneic HSCTs (allo-HSCTs) and 1625 autologous HSCTs (auto-HSCTs) among 54 transplant centers during 2014 (SIGNB-GITMO-AMCLI study). Using logistic regression methods. we identified risk factors for GNB and evaluated the impact of GNB on the 4-month overall-survival after transplant. Results The cumulative incidence of pre-engraftment GNB was 17.3% in allo-HSCT and 9% in auto-HSCT. Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa were the most common isolates. By multivariate analysis, variables associated with GNB were a diagnosis of acute leukemia, a transplant from a HLA-mismatched donor and from cord blood, older age, and duration of severe neutropenia in allo-HSCT, and a diagnosis of lymphoma, older age, and no antibacterial prophylaxis in auto-HSCT. A pretransplant infection by a resistant pathogen was significantly associated with an increased risk of posttransplant infection by the same microorganism in allo-HSCT. Colonization by resistant gram-negative bacteria was significantly associated with an increased rate of infection by the same pathogen in both transplant procedures. GNB was independently associated with increased mortality at 4 months both in allo-HSCT (hazard ratio, 2.13; 95% confidence interval, 1.45–3.13; P Conclusions Pre-engraftment GNB is an independent factor associated with increased mortality rate at 4 months after auto-HSCT and allo-HSCT. Previous infectious history and colonization monitoring represent major indicators of GNB. Clinical Trials registration NCT02088840.

Nonostante la prognosi dei bambini con leucemia acuta mieloide (LAM) sia migliorata in modo significativo nel corso degli ultimi 30 anni, circa il 30% dei bambini recidiva, facendo di quest’ultima la principale causa di fallimento terapeutico e di morte. Per indagare i meccanismi molecolari alla base recidiva, è stato eseguito il sequenziamento massivo dell’esoma dei campioni di esordio, remissione e recidiva di 4 LAM pediatriche a citogenetica normale mediate tecnologia Illumina, seguito da sequenziamento mirato high coverage (7000X) delle mutazioni somatiche con possibile ruolo patogenetico nello sviluppo della recidiva. La mutazione biCEBPα è stabile e altamente penetrante durante il decorso della malattia (>80% nel clone di esordio e recidiva). Al contrario, le mutazioni di WT1 risultano estremamente instabili. Si configurano specifici pattern molecolari sottostanti alla recidiva, tra i quali l’aberrante attivazione dei segnali proliferativi cellulari (conferito dalle mutazioni di PTPN11 e FLT3-TKD) e l’aumentata resistenza all’apoptosi (iperattivazione di TYK2). Si osserva inoltre un’instabilità genomica conferita dall’inattivazione di SETD2, una metiltransferasi implicata nel mismatch repair, alla base di una maggior plasticità della malattia che contribuisce alla sua evoluzione. Il conseguente accumulo di nuove mutazioni promuove l'adattabilità della leucemia, contribuendo alla selezione clonale. E’ stata inoltre identificata una nuova mutazione di ASXL3, presente in un clone minoritario alla diagnosi (, Despite significant improvement in treatment of childhood acute myeloid leukemia (AML), 30% of patients experience disease recurrence, which is still the major cause of treatment failure and death in these patients. To investigate molecular mechanisms underlying relapse, we performed whole-exome sequencing of diagnosis-relapse pairs and matched remission samples from 4 pediatric AML patients without recurrent cytogenetic alterations. Candidate driver mutations were selected for targeted deep sequencing at high coverage, suitable to detect small subclones (0.12%). BiCEBPa mutation was found to be stable and highly penetrant, representing a separate biological and clinical entity, unlike WT1 mutations, which were extremely unstable. Among the mutational patterns underlying relapse, we detected the acquisition of proliferative advantage by signaling activation (PTPN11 and FLT3-TKD mutations) and the increased resistance to apoptosis (hyperactivation of TYK2). We also found a previously undescribed feature of AML, consisting of a hypermutator phenotype caused by SETD2 inactivation. The consequent accumulation of new mutations promotes the adaptability of the leukemia, contributing to clonal selection. We report a novel ASXL3 mutation characterizing a very small subclone (

Climate changes resulting from increases in temperature and air pollution influence pollen allergenicity, responsible for the dramatic raise in respiratory allergies. Allergies are a complex of symptoms derived from altered IgE-mediated reactions of the immune system towards substances known as allergens. Allergic sensibilization can be of food or respiratory origin; many allergens have been identified in pollens or fruits of different plant species and allergic cross-reactivity can occur in a patient reacting to similar allergens from different origins. Although the link between climate change and pollen allergenicity is proven, the underlying mechanism is little understood. Transglutaminases (TGases), enzymes able to post-translationally modify proteins, are activated under stress and involved in some inflammatory responses, enhancing the activity of pro-inflammatory phospholipase A2, (PLA2). A calcium-dependent TGase activity has been identified in the apple pollen cell wall, raising the possibility that TGase may have a role in the modification of allergens and, when pollen arrives in contact with the human mucosa, the enzyme could be responsible in the transamidation of proteins as PLA2. In Europe a great number of people suffers from apple fruit allergy, but little is known on fruit/pollen cross-allergenicity. Apple fruit allergies are mainly due to four different classes of allergens: Mal d 1, 2, 3, 4. Here we have investigated the presence of Mal d allergens also in pollen to clarify its allergenic potential using a RT-PCR approach both on ungerminated and germinated pollen and we show our initial effort on clarification of climate changes effects on the expression of Mal d allergens. This multidisciplinary approach, aimed to establish the scientific basis for future studies on pollen-food cross allergenicity is supported by the grant “Crossallergenicity” Progetto Strategico d’Ateneo E.F. 2006, University of Bologna.

Le proteine allergeniche della mela presentano delle caratteristiche comuni tra loro: - Tutte le proteine allergeniche sono codificate da famiglie geniche composte da un numero variabile di geni, gli isoallergeni, di cui per ognuna esitono numerose varianti alleliche. Questa presenza ridondante di sequenze Mal d nel genoma del melo, ed in particolare su cromosomi omologhi, sembra essere dovuta alla natura allopoliploide del melo e questo fa anche supporre che il numero di isoallergeni noti sia destinato a crescere ulteriormente. - Tre classi di proteine su quattro (Mal d 1, Mal d 2 e Mal d 3) appartengono alle proteine PR e quindi sono in qualche modo correlate alla difesa della pianta da vari tipi di stress, sia biotici che abiotici. - Alcuni fattori comuni a molti allergeni come la stabilità a bassi pH, alle alte temperature e a proteolisi o la capacità di trasportare lipidi o altri ligandi potrebbero influenzarne l’allergenicità. Nonostante l’aumento delle conoscenze in questi ultimi anni, non è stato ancora chiarito quali isoallergeni siano effettivamente espressi nel frutto e quali siano i veri responsabili della reazione allergica. Questo è un punto chiave nel problema delle allergie alla mela ma anche alla frutta e agli ortaggi in generale. Nella impostazione della ricerca su cui abbiamo qui riferito, si è scelto un approccio multidisciplinare che prevede l’integrazione, a fianco delle biotecnologie, di discipline come l’allergologia e la pomologia (Figura 8). Attraverso questi studi si tenterà di associare ad ogni locus genico un determinato potenziale allergenico e quindi, individuando le sequenze di isoallergeni associate ad un basso grado di allergenicità, si potrà anche orientare il miglioramento genetico del melo verso l’ottenimento di varietà a bassa allergenicità. Una volta individuati dei marker per la bassa allergenicità, si potranno cercare genotipi interessanti per questo carattere anche esplorando le collezioni dell’antico germoplasma del melo. Un aspetto però da non sottovalutare è la doppia faccia della medaglia ovvero il ruolo delle proteine responsabili dell’allergenicità; molte di queste, infatti, sono coinvolte nella risposta della pianta ad attacchi di patogeni e altri stress. Questa considerazione rende inevitabile la preoccupazione che l’auspicata riduzione dell’allergenicità, possa comportare una maggiore suscettibilità agli stress indesiderabile per ovvi motivi. Per ora è solamente un interrogativo. Considerando i notevoli effetti benefici delle mele sulla salute degli individui e considerando il notevole interesse che, negli ultimi anni, il mercato riserva alla nutraceutica e agli alimenti correlati al benessere, non è da escludere che lo studio del carattere riguardante l’ipoallergenicità dei frutti possa essere interessante non solo per quanto riguarda la salute umana ma anche per un possibile riscontro economico

Allergies are a complex of symptoms derived from altered IgE-mediated reactions of the immune system towards substances known as allergens. Apple and hazelnut allergens have been identified in pollens or fruits. Apple allergies are due to four different classes of allergens (Mal d 1, 2, 3, 4), whose allergenicity is related both to genotype and tissue specificity. Building by Homology technique was applied to predict the 3D structures of Mal d 1-4. Starting from the comparison of these results leading to understand the different ways of reactions of the Malds, we have studied the answer of sensitised individuals to 11 apple genotypes, separating fruits skin positive answers from those to the pulp through Skin Prick Test and prick-to-prick test, evidencing different allergic responses to the different cultivar. Climate changes resulting from increases in temperature and air pollution influence pollen allergenicity, responsible for the dramatic raise in respiratory allergies (hay fever, bronchial asthma, conjunctivitis). Although the link between climate change and pollen allergenicity is proven, the underlying mechanism is little understood. Transglutaminases (TGases), a class of enzymes able to post-translationally modify proteins, are activated under stress and involved in some inflammatory responses, enhancing the activity of pro-inflammatory phospholipase A2, suggesting a role in allergies. Aero allergenic (hazelnut. Corylus avellana) and enthomophylus (apple. Malus domestica) pollen exposed to climate changes (different temperatures, relative humidity (rH), acid rain at pH 5.6 and copper pollution (3.10 µg/l)) showed an increase in pollen surface TGase activity in Corylus, while Malus pollen TGase was not significantly affected. Several pollen proteins were post-translationally modified, including the activation of mammalian sPLA2, thus potentially altering pollen allergenicity and inflammation. Western blotting of pollen TGase revealed its induced expression after exposure to stressors, while detection of in situ TGase activity and morphological examination indicated pollen damage and release of TGase. Grants:Crossallergenicity Progetto Strategico d’Ateneo E.F. 2006,University of Bologna

Premessa/obiettivi: Sono state descritte 4 famiglie di allergeni in melo (Mal d 1, 2, 3, 4 [1]) e sintomi clinici associati [2-3]. Un diverso grado di allergenicità è riportato in letteratura sia in relazione al genotipo [4] che al tessuto analizzato: alcuni allergeni sembrano localizzarsi soprattutto a livello della buccia [5]. Scopo dello studio è determinare la risposta di pazienti allergici alla mela attraverso SPT. Materiali e Metodi: 19 soggetti, giunti consecutivamente presso l’Ambulatorio di Allergologia Pediatrica con allergia alimentare alla mela (prevalentemente come sindrome orale allergica), sono stati sottoposti a SPT per estratto di mela (Lofarma) e per 11 genotipi di melo (Florina, Durello, Ohrin, Red Chief, Jonagold, Jonathan, Topaz, Fiesta, Fuji, Gala, Golden), condotti separatamente per buccia e polpa (prick-to-prick). Risultati: Sono stati considerati positivi pomfi di diametro maggiore-uguale a quello dell’istamina (hys) dello stesso paziente. Le polpe di Durello e Ohrin sono risultate meno sensibilizzanti, causando percentualmente la minore positività (rispettivamente 54.5% e 57.9%) e una reazione mediamente inferiore a hys (SPT/hys < 1). Invece, la maggior parte dei pazienti ha mostrato una forte reazione alla polpa di Jonathan e Jonagold (positività > 73.7%, SPT/hys > 1,36). Le bucce di Durello e Jonagold mostrano una minore reazione, rispettivamente con percentuale di positività di 54.5% e 57.9% e SPT/hys di 0.71 e 1.0. Una forte reazione è invece presente per i genotipi Gala (positività = 94.7%, SPT/hys = 1.43) e Fiesta (positività = 78.9%, SPT/hys = 1.46). La somma del numero di positività dei pomfi risultati dalla buccia (144) è lievemente maggiore rispetto al numero di positività della polpa (136). Tuttavia, esiste una variabilità di risposta fra polpa e buccia in diversi genotipi, specie per Ohrin: dei 18 soggetti che hanno presentato almeno 1 positività tra polpa e buccia, solo in 8 casi entrambi i tessuti erano positivi. Discussione: I risultati suggeriscono che vi siano differenze allergeniche nelle diversi cultivar, tuttavia ciò è complicato dalla variabilità legata alla risposta dei pazienti. Bibliografia: 1. Fernandez-Rivas e coll. JACI 2006; 118: 481-8. 2. Marzban e coll. Plant Science 2005; 169: 387-94. 3. Sancho e coll. Allergy 2005;60: 1262-8. 4. Bolhaar e coll. JACI 2005;116: 1080-6. 5. Borges e coll. Plant Physiology and Biochemistry 2006;44: 535-42.

The gut microbiome (GM) has shown to influence hematopoietic stem cell transplantation (HSCT) outcome. Evidence on levofloxacin (LVX) prophylaxis usefulness before HSCT in pediatric patients is controversial and its impact on GM is poorly characterized. Post-HSCT parenteral nutrition (PN) is oftentimes the first-line nutritional support in the neutropenic phase, despite the emerging benefits of enteral nutrition (EN). In this exploratory work, we used a global-to-local networking approach to obtain a high-resolution longitudinal characterization of the GM in 30 pediatric HSCT patients receiving PN combined with LVX prophylaxis or PN alone or EN alone. By evaluating the network topology, we found that PN, especially preceded by LVX prophylaxis, resulted in a detrimental effect over the GM, with low modularity, poor cohesion, a shift in keystone species and the emergence of modules comprising several pathobionts, such as Klebsiella spp., [Ruminococcus] gnavus, Flavonifractor plautii and Enterococcus faecium. Our pilot findings on LVX prophylaxis and PN-related disruption of GM networks should be considered in patient management, to possibly facilitate prompt recovery/maintenance of a healthy and well-wired GM. However, the impact of LVX prophylaxis and nutritional support on short- to long-term post-HSCT clinical outcomes has yet to be elucidated.

Summary Background Presenting symptoms of childhood cancers might mimic those of rheumatic diseases. However, the evidence available to guide differential diagnosis remains scarce. Preventing wrong or delayed diagnosis is therefore important to avoid incorrect administration of glucocorticoid or immunosuppressive therapy and worsening of prognosis. As such, we aimed to assess the prevalence and characteristics of presenting musculoskeletal manifestations in patients at cancer onset and to identify the factors that differentiate childhood malignancies with arthropathy from juvenile idiopathic arthritis. Methods We did a multicentre, cross-sectional study at 25 paediatric haemato-oncology centres and 22 paediatric rheumatology centres in Italy. We prospectively recruited patients who were younger than 16 years that were newly diagnosed with cancer or juvenile idiopathic arthritis. We excluded patients with glucocorticoid pre-treatment (>1 mg/kg per day of oral prednisone or equivalent for ≥2 consecutive weeks). We collected data for patients with a new diagnosis of cancer or juvenile idiopathic arthritis using an electronic case report form on a web-based platform powered by the Cineca Interuniversity Consortium. The primary outcome was to describe the frequency and characteristics of musculoskeletal manifestations at cancer onset; and the secondary outcome was to identify factors that could discriminate malignancies presenting with arthropathy, with or without other musculoskeletal symptoms, from juvenile idiopathic arthritis using multivariable logistic regression analysis. Findings Between May 1, 2015, and May 31, 2018, 1957 patients were eligible, of which 1277 (65%) had cancer and 680 (35%) had juvenile idiopathic arthritis. Musculoskeletal symptoms occurred in 324 (25% [95% CI 23·0–27·8]) of 1277 patients with cancer, of whom 207 had arthropathy. Patients with malignant bone tumours had the highest frequency of musculoskeletal symptoms (53 [80%] of 66), followed by patients with Langerhans histiocytosis (16 [47%] of 34), leukaemia (189 [32%] of 582), soft-tissue sarcomas (16 [24%] of 68), and neuroblastoma (21 [19%] of 109). In the 324 patients with cancer and musculoskeletal symptoms, the most common complaints were joint pain (199 [61%]), followed by limb bone pain (112 [35%]). Joint involvement had a prevalent monoarticular pattern (100 [48%] of 207) and oligoarticular pattern (86 [42%] had 2–4 joints involved and 20 [10%] had >4 joints involved), with the most frequently involved joints being the hip (88 [43%] of 207) and knee (81 [39%]). On multivariable analysis, limb bone pain was the independent variable most strongly associated with cancer (odds ratio [OR] 87·80 [95% CI 18·89–408·12]), followed by weight loss (59·88 [6·34–565·53]), thrombocytopenia (12·67 [2·40–66·92]), monoarticular involvement (11·30 [4·09–31·19]), hip involvement (3·30 [1·13–9·61]), and male sex (2·40 [1·03–5·58]). Factors independently associated with juvenile idiopathic arthritis were morning stiffness (OR 0·04 [95% CI 0·01–0·20]), joint swelling (0·03 [0·01–0·09]), and involvement of the small hand joints (0·02 [0–1·05]). Interpretation Our study provides detailed information about presenting musculoskeletal manifestations of childhood cancers and highlights the clinical and laboratory features that are most helpful in the differential diagnosis with juvenile idiopathic arthritis. Funding Associazione Lorenzo Risolo.

Oral chronic graft versus host disease (cGVHD) is often refractory to systemic therapies. Additional topical treatment is commonly required. The potency of the agent, the vehicle and formulation in which it is delivered are all critical factors in determining the effectiveness of topical therapies. High potency of budesonide, combined with its very low bioavailability when absorbed through mucosal surfaces, increased the potential role in topical application for oral cGVHD. Viscous formulation increases mucosal contact time resulting in a greater decrease in mucosal inflammation. This short communication suggests that oral viscous budesonide should be considered as a potential new therapy in the management of oral cGVHD.

// Riccardo Masetti 1, * , Ilaria Castelli 1, * , Annalisa Astolfi 2 , Salvatore Nicola Bertuccio 1 , Valentina Indio 2 , Marco Togni 1, 3 , Tamara Belotti 1 , Salvatore Serravalle 1 , Giuseppe Tarantino 2 , Marco Zecca 4 , Martina Pigazzi 5 , Giuseppe Basso 5 , Andrea Pession 1, # , Franco Locatelli 6, 7, # 1 Department of Pediatrics “Lalla Seragnoli”, Hematology-Oncology Unit, University of Bologna, Bologna, Italy 2 Interdepartmental Centre of Cancer Research “G. Prodi”, University of Bologna, Bologna, Italy 3 Current address: Stem Cell Group, University College London Cancer Institute, University College London, London, United Kingdom 4 Department of Pediatric Hematology Oncology, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy 5 Department of Woman and Child Health, Laboratory of Hematology-Oncology, University of Padova, Padova, Italy 6 Department of Pediatric Hematology-Oncology, IRCCS Ospedale Bambino Gesu, Rome, Italy 7 University of Pavia, Pavia, Italy * These authors have contributed equally to this work # Both authors have shared co-senior authorship Correspondence to: Riccardo Masetti, email: riccardo.masetti@gmail.com Keywords: pediatric acute myeloid leukemia, acute myeloid leukemia relapse, whole-exome massively parallel sequencing, SETD2 mutation, FLT3-TKD mutation Received: April 19, 2016 Accepted: July 10, 2016 Published: July 22, 2016 ABSTRACT Despite significant improvement in treatment of childhood acute myeloid leukemia (AML), 30% of patients experience disease recurrence, which is still the major cause of treatment failure and death in these patients. To investigate molecular mechanisms underlying relapse, we performed whole-exome sequencing of diagnosis-relapse pairs and matched remission samples from 4 pediatric AML patients without recurrent cytogenetic alterations. Candidate driver mutations were selected for targeted deep sequencing at high coverage, suitable to detect small subclones (0.12%). BiCEBPα mutation was found to be stable and highly penetrant, representing a separate biological and clinical entity, unlike WT1 mutations, which were extremely unstable. Among the mutational patterns underlying relapse, we detected the acquisition of proliferative advantage by signaling activation (PTPN11 and FLT3-TKD mutations) and the increased resistance to apoptosis (hyperactivation of TYK2). We also found a previously undescribed feature of AML, consisting of a hypermutator phenotype caused by SETD2 inactivation. The consequent accumulation of new mutations promotes the adaptability of the leukemia, contributing to clonal selection. We report a novel ASXL3 mutation characterizing a very small subclone (