Genetic studies of susceptibility to inflammation, autoimmunity, and hematological malignancy

The immune system represents the body’s defense against infectious organisms. Inborn defects of the immune system, called primary immunodeficiencies (PIDs), are a heterogeneous group of Mendelian disorders. Clinically, PIDs can cause isolated to broad susceptibility to pathogens, severe hyperinflammation, autoimmunity, allergy, and cancer. The studies in this thesis take advantage of the recent development in DNA-sequencing technologies and of our increased understanding of genetic variability to further explore the genetic architecture and phenotypic spectrum of hemophagocytic lymphohistiocytosis (HLH), an inborn error of lymphocyte cytotoxicity, and to elucidate the genetic factors behind autoimmunity and hematological malignancies in selected families. Familial HLH (FHL) is a severe hyperinflammatory condition, genetically heterogeneous, caused by defective perforin-mediated lymphocyte cytotoxic activity. In paper I we use a highthroughput sequencing panel covering 12 HLH-related genes in 58 prospectively recruited patients with HLH and achieve a molecular diagnosis in 22 cases (38%). In paper II we show that perforin-deficiency due to biallelic PRF1 missense variants is associated with broad intrafamilial variability and clinical presentations seemingly unrelated to HLH, such as Hodgkin lymphoma. Using exome sequencing, in paper III, we identify biallelic pathogenic variants in IFNGR1 and INFGR2, respectively, in two patients with HLH and disseminated mycobacterial infection. Previous studies have shown that HLH pathology is largely driven by IFN-γ. Instead, our findings suggest the existence of IFN-γ-independent mechanisms for the development of HLH. In paper IV, we uncover biallelic coding and non-coding variants in RAB27A, the gene responsible for Griscelli syndrome type 2 (GS2), in five unrelated patients with atypical HLH, normal pigmentation, and a functional defect suggestive of FHL. A complex structural variant disrupting the transcriptional start site (TSS) of one RAB27A transcript was shared among the patients. We show that the disrupted TSS is less predominantly used by melanocytes compared to lymphocytes, explaining the lack of hypopigmentation in these patients, otherwise present in GS2. In paper V we report the beneficial effect of hematopoietic stem cell transplantation in a 14-yearold boy with LRBA deficiency and a seven-year history of severe autoimmmune disorders. In paper VI we uncover germline heterozygous missense variants in SAMD9L, a gene located on 7q21, in two families with multiple individuals affected by cytopenia, immunodeficiency, myelodysplastic syndrome (MDS) with cytogenetic aberrations of chromosome 7, and neurological disease. We show a gain-of-function (GOF) effect of the mutants, which inhibit cell proliferation. Germline SAMD9L GOF variants were lost in MDS cells and hematopoietic revertant mosaicism occurred frequently among less severely affected carriers. Our results indicate a strong selective advantage for hematopoietic cells that, through different somatic events, overcome the growth-inhibiting effect of germline SAMD9L GOF variants. Taken together, these studies add to our understanding of the phenotypic and genetic spectrum of HLH, display the power of high-throughput sequencing in diagnostics of individuals affected by severe inflammation, autoimmunity, and hematological malignancies, and highlight SAMD9L as an important gene for regulation of hematopoietic cell proliferatio

A hemophagocytic lymphohistiocytosis case with newly defined UNC13D (C.175G>C; p.Ala59Pro) mutation and a rare complication

Hemophagocytic lymphohistiocytosis (HLH) represents a severe hyperinflammatory condition with cardinal symptoms of prolonged fever, cytopenias, hepatosplenomegaly, and hemophagocytosis by activated, morphologically benign macrophages with impaired function of natural killer cells and cytotoxic T lymphocytes. A 2-month-old girl, who was admitted with fever, was diagnosed with HLH and her genetic examination revealed a newly defined mutation in the UNC13D (c.175G>C; p.Ala59Pro) gene. She was treated with dexamethasone, etoposide, and intrathecal methotrexate. During the second week of treatment, after three doses of etoposide, it was noticed that there was a necrotic plaque lesion on the soft palate. Pathologic examination of debrided material in PAS and Grocott staining revealed lots of septated hyphae, which was consistent with aspergillosis infection. Etoposide was stopped and amphotericin B treatment was given for six weeks. HLH 2004 protocol was completed to eight weeks with cyclosporine A orally. There was no patient with invasive aspergillosis infection as severe as causing palate and nasal septum perforation during HLH therapy. In immuncompromised patients, fungal infections may cause nasal septum perforation and treatment could be achieved by antifungal therapy and debridement of necrotic tissue. © 2015 Turkish Society of Hematology. All rights reserved

Variability of forebrain commissures in callosal agenesis: a prenatal magnetic resonance imaging study

Introduzione e Scopo dello Studio L’agenesia completa del corpo calloso (ACC) anche quando isolata può essere espressione di eterogeneità anatomica. Lo scopo dello studio è descrivere la variabilità delle altre strutture commissurali in un’ ampia corte di feti con apparente agenesia completa isolata del corpo calloso con risonanza magnetica (RMN) in epoca pretale Materiale e Metodi In tutti i feti con a ACC riferiti all’ analisi con RM dal 2004 al 2014 presso la Neuroradiologia Pediatrica dell’ Ospedale Buzzi di Milano è stata valutata, da parte di due neuroradiologi esperti con più di 10 anni di esperienza in questo campo, la presenza di altre strutture commissurali: la commissura anteriore (CA) e la commissura ippocampale (CI) Risultati Complessivamente sono stati reclutati 62 casi: 3/62 (4,8%) non presentavano nessuna struttura commissurale cerebrale; 23/62 (37,1%) presentavano solo la CA, 20/62 (32,3%) presentavano sia la CA che la CI in una forma vestigiale, mentre i rimanenti 16/62 casi evidenziavano la presenza di una commissura ibrida (CIB) risultanza della fusione della CI vestigiale con un corpo rudimentale e prematuro del corpo calloso. I reperti prenatali sono stati successivamente confermati dalle immagini ottenute con la RM post-natale quando disponibili. Conclusioni Nella maggioranza dei feti della nostra corte era stata documentata allo studio prenatale con RM la presenza almeno di una commissura (CA); in circa la metà di essi era stata identificata in concomitanza la presenza di una seconda commissura: CI in una forma vestigiale o la CIB composta dalla fusione di due commissure; la CI vestigiale e il prematuro corpo del corpo calloso. Ulteriori studi sono necessari al fine di verificare se tale variabilità commissurale nei feti affetti da ACC isolata possa associarsi a diverse caratteristiche genetiche e avere un impatto sull’ outcome neurologico a lungo termine di questi giovani pazienti.Background and Purpose Agenesis of corpus callosum (ACC), even when isolated, may be characterized by anatomical variability. The aim of this study was to describe the types of other forebrain commissures, in a large cohort of randomly enrolled fetuses with apparently isolated ACC at prenatal magnetic resonance (MR) imaging. Materials and Methods From 2004 to 2014 in all fetuses with apparently isolated ACC submitted to prenatal MR imaging, the presence of the anterior (AC) or a vestigial hippocampal commissure (HC) was assessed "in consensus" by two pediatric neuroradiologists. Results Overall 62 cases of ACC were retrieved from our database. In 3/62 fetuses (4,8%) no forebrain commissure was visible at prenatal MR imaging, 23/62 cases (37,1%) presented only the AC, 20/62 cases (32,3%) showed both the AC and a residual vestigial HC, whereas in the remaining 16/62 cases (25,8%) a hybrid structure (HS) merging a residual vestigial HC and a rudiment of CC body was detectable. Postnatal MR imaging, when available, confirmed prenatal forebrain commissure findings. Conclusions The vast majority of fetuses with apparently isolated ACC showed at least one forebrain commissure at prenatal MR imaging, and about half of cases also a second commissure: a vestigial HC or a hybrid made of HC and rudiment CC body. It remains to be assessed if such variability is the result of different genotype and if it may have any impact on the long term neurodevelopmental outcom

La nozione della vendita: causa e oggetto

Analisi sistematica della causa e dell'oggetto del contratto di compravendit

Influenza di diversi pre-trattamenti sulle cinetiche di disidratazione di olive

In this paper, the drying behaviour of three olive cultivars (Olea europea sativa Hoffm. and Link), was studied. In particular, the influence of different pre-treatments on dehydration kinetics was investigated. Olives were calibrated and subjected to several preliminary treatments, in order to break the peel, before being dehydrated. Dehydration was carried out at 60°C with constant flow rate and humidity. Fruits were placed on trays according to a completely randomised design. Dehydration kinetics were computed by weighing fruits during dehydration. Results show that the dehydration rate was drastically increased by piercing the fruit cuticle, while the others systems did not give appreciable beneficial effects. Dried olives were appreciated by panellists. In questo lavoro è stata studiata la risposta alla disidratazione di tre varietà di olive (Olea europea sativa Hoffm. and Link). In particolare, si è verificata l’influenza di diversi pre-trattamenti sulle cinetiche di disidratazione. Le olive, dopo la calibratura, sono state sottoposte a diversi pre-trattamenti, nell’intento di creare delle microlesioni superficiali, poi disidratate in un armadio con aria a flusso ed umidità costanti alla temperatura di 60°C, disponendo i frutti delle tesi secondo un piano completamente randomizzato. Ad intervalli prestabiliti durante la disidratazione sono stati effettuati dei prelievi di frutti, al fine di determinarne le cinetiche. I risultati mostrano che la tecnica della scarificatura superficiale ha incrementato la velocità di disidratazione, mentre gli altri trattamenti non hanno dato risultati degni di nota. I frutti disidratati sono stati giudicati favorevolmente dagli assaggiatori

La nozione della vendita: gli effetti traslativi

Analisi della disciplina degli effetti traslativi nel contratto di compravendita con particolare riguardo al principio del consenso traslativ

Pendenza della condizione. Aspettativa di diritto. Atti di disposizione

La situazione giuridica di aspettativa sia caratterizzata da due elementi essenziali: uno, sostanziale, rappresentato dalla necessaria presenza di un interesse alla conservazione, in vista della sua realizzazione, del contenuto negoziale; e uno formale, consistente nell'insieme di poteri, di natura conservativa e strumentale, posti a garanzia del raggiungimento del risultato avuto di mira. Il problema di fondo è quello di verificare se e in che misura sia possibile configurare una situazione di aspettativa tutte le volte in cui si riscontri la esistenza di una pendenza giuridicamente tutelata. La presenza di effetti preliminari e conservativi non sempre assurge a situazione giuridica di aspettativa. La situazione giuridica di aspettativa non è ricostruibile come categoria unitaria, né come diritto soggettivo autonomo; l’esigenza di conservazione gode di una protezione diversamente modulata a seconda della fattispecie a cui è correlata e, soprattutto, della particolare natura degli interessi in gioco che determinano la pendenza.The legal situation of expectation is characterized by two essential elements: one, substantial, represented by the necessary presence of an interest in the preservation to its realization of the negotiating content ;and a formal one, consisting of a set of powers of a conservative and instrumental nature, designed to guarantee the achievement of the interest pursued. The problem is to verify if and to what extent it is possible to configure an expectation situation every time that there is evidence of the existence of a legally protected slope.The presence of preliminary and conservative effects does not always lead to a legal situation of expectation. The legal situation of expectation can not be reconstructed as a unitary category, nor as an autonomous subjective right; the need for conservation enjoys a differently modulated protection depending on the case to which it is related and, above all, on the particular nature of the interests involved that determine the slope

Els noms dels colors del pelatge del bestiar en la documentació de l'Alguer dels segles XVIII al XIX

A partir de la tesi de doctorat 'La interferència dels parlars sards en el català de l'Alguer entre els segles XVII i XVIII. Estudi del lèxic a través dels Registres de danys de la Barracelleria /1683-1784)', l'article dóna compte del grau d'interferència lèxica del sard en el camp lexicosemàntic dels noms dels colors pelatge del bestiar i d'altres característiques físiques, un camp altament interferit pel sard, com demostren els documents analitzats dels segles XVIII i XIX (Registres de cavalls i de bísties de mola, Registres de danys, Registres de bestiar viu)