The formation of blood, hematopoiesis, is a dynamic process originating from a small number of hematopoietic stem cells (HSCs). To sustain hematopoiesis throughout life HSCs have the unique capacity to differentiate into all mature hematopoietic lineages as well as generating more HSCs by a mechanism referred to as self-renewal. However, the regulation of these processes is largely unknown. During embryonic development HSCs expand in the fetal liver, indicating that this environment supports HSC self-renewal. The LIM-homeobox gene Lhx2 is expressed in the fetal liver during this period and Lhx2 null mutant mice die in utero due to severe anemia caused by an environmental defect in the fetal liver. Embryonic stem cells differentiate in vitro, forming embryoid bodies (EBs) containing various tissues including hematopoietic progenitor cells. Introduction of Lhx2 into this system by retroviral transfer led to the generation of cytokine dependent HSC-like cell lines that were multipotent and expressed surface markers similar to embryonic HSCs. However, the specificity and efficiency of this event could not be elucidated. To further evaluate the function of Lhx2 expression during hematopoietic development, Lhx2 was introduced into an ES cell system where expression could be efficiently turned on. This approach revealed that Lhx2 induce self-renewal of distinct multipotent hematopoietic progenitor/stem cells present in the EB, with the ability to form HSC-like cell lines. The Lhx2 induced self-renewal is growth factor specific since stem cell factor and interleukin-6 are necessary and sufficient for this process. However, Lhx2 expression blocked erythroid differentiation and interfered with early ES cell commitment, indicating that the effect of Lhx2 is cell type specific. Since HSCs of early embryonic origin are inefficient in engrafting adult recipients upon transplantation, we wanted to address whether we could generate cell lines retaining this capacity by expression of Lhx2 in hematopoietic cells from adult bone marrow. This led to the generation of clonal and cytokine dependent HSC-like cell lines capable of generating erythroid, myeloid and lymphoid cells upon transplantation into lethally irradiated recipients. When transplanted into stem cell-deficient mice, they contributed to circulating erythrocytes for at least 18 months, revealing a remarkable potential for self-renewal and differentiation in vivo. However, expression of Lhx2 was maintained in vivo and most engrafted mice developed a transplantable myeloproliferative disorder resembling human chronic myeloid leukemia. Thus, elucidation of the mechanism for Lhx2 function in HSC-like cell lines would give insights into both normal and pathological regulation of HSCs. Down-regulation of Lhx2 expression in HSC-like cell lines with inducible Lhx2 expression led to rapid loss of stem cell characteristics and differentiation into various hematopoietic cell types. Thus, global gene expression analysis comparing Lhx2+ HSC-like cell lines to their Lhx2- progeny would give insights into the molecular basis for Lhx2 function in stem cells. A number of differentially expressed genes overlapped with previously reported HSC enriched genes, further emphasizing the resemblance between HSCs and the HSC-like cell lines also at the molecular level. Moreover, a number of genes were identified with functions or expression patterns related to Lhx2 in other organs. Collectively, these data suggest that these HSC-like cell lines represent a relevant model system for normal HSCs on the molecular and the functional level as well as for evaluating Lhx2 function in the development of various tissues in the embryo as well as in disease.

We are unceasingly exposed to potentially harmful microorganisms. The battle against threatening infectious agents includes activation of both the innate and of the adaptive immune systems. Illness responses are elicited and include inflammation, fever, decreased appetite, lethargy and increased sensitivity to painful stimuli in order to defeat invaders. While many of these signs of disease are controlled by the central nervous system, it has remained an enigma how signals from the peripheral immune system reach the brain through its blood-brain barrier, which precludes macromolecules, including cytokines, from diffusing into the brain parenchyma. Previous findings indicate the existence of a pathway across the blood-brain barrier, which includes binding of the cytokine interleukin-1 (IL-1) to its receptor in the brain vessels, thereby inducing the production of the prostaglandin E2 (PGE2) synthesizing enzymes cyclooxygenase-2 (Cox-2) and microsomal prostaglandin E synthase-1 (mPGES-1), which ultimately synthesize PGE2. PGE2 subsequently binds to any of the four prostaglandin E2 (EP) -receptors. Previous results from our laboratory have suggested that this pathway plays a critical role in the febrile response to infectious stimuli. The present thesis aims at further investigating the molecular events underlying immune-to-brain signalling, with special emphasis on fever, hypothalamic-pituitary-adrenal (HPA) -axis activation and anorexia and their connection to signalling molecules of the cytokine and prostaglandin families, respectively. In paper I, the molecular processes linking the proinflammatory cytokine interleukin-6 (IL-6) and PGE2 in the febrile response were investigated. Both IL-6 and PGE2 have been shown to be critical players in the febrile response, although the molecular connections are not known, i.e. if IL-6 exerts its effects up- or downstream of PGE2. Mice deficient in IL-6 were unable to respond to bacterial lipopolysaccharide (LPS) with a febrile response, but displayed similar induction of Cox-2 and mPGES-1, and similar concentrations of PGE2 in the cerebrospinal fluid as wild-type mice. Paradoxically, the IL-6 deficient mice responded with a dose-dependent elevation of body temperature in response to intracerebroventricularly injected PGE2. Furthermore, IL-6 per se was not pyrogenic when injected peripherally in mice, and did not cause increased levels of PGE2 in cerebrospinal fluid. IL-6 deficient mice were not refractory to the action of PGE2 because of excess production of some hypothermia-producing factor, since administration of a Cox-2 inhibitor in LPS-challenged IL-6 deficient mice did not unmask any hypothermic response, and neutralization of tumor necrosis factor α (TNFα), associated with hypothermia, did not produce fever in LPS-challenged IL-6 deficient mice. These data indicate that IL-6 rather than exerting its effects up- or down-stream of PGE2 affects some process in parallel to PGE2, perhaps by influencing the diffusion and binding of PGE2 onto its target neurons. In papers II and III, we injected the proinflammatory cytokine IL-1β in free-fed wild-type mice, in mice with a deletion of the gene encoding mPGES-1, or in mice deficient in the EP1, EP2 and EP3. Food intake was continuously measured during their active period, revealing that mPGES-1 deficient mice were almost completely resistant to anorexia induced by IL-1β. However, all of the investigated EP receptor deficient mice exhibited a normal profound anorexic response to IL-1β challenge, suggesting that the EP4 is the critical receptor that mediates IL-1β-induced anorexia. We also investigated the role of mPGES-1 in anorexia induced by lipopolysaccharide (LPS) in mPGES-1 deficient mice. The profound anorexic response after LPS-challenge was similar in mPGES-1 deficient and wild-type mice. To further investigate the anorectic behaviour after LPS injection, we pre-starved the animals for 22 hours before injecting them with LPS. In this paradigm, the anorexia was less profound in mPGES-1 knock-out mice. Our results suggest that while the inflammatory anorexia elicited by peripheral IL-1β seems largely to be dependent on mPGES-1-mediated PGE2 synthesis, similar to the febrile response, the LPS-induced anorexia is independent of this mechanism in free-fed mice but not in pre-starved animals. In papers IV and V, the role of prostanoids for the immune-induced HPA-axis response was investigated in mice after genetic deletion or pharmacological inhibition of prostanoid-synthesizing enzymes, including Cox-1, Cox-2, and mPGES-1. The immediate LPS-induced release of ACTH (adrenocorticotropic hormone and corticosteroids was critically dependent on Cox-1 derived prostanoids and occurred independently of Cox-2 and mPGES-1 derived PGE2. In contrast, the delayed HPA-axis response was critically dependent on immune-induced PGE2, synthesized by Cox-2 and mPGES-1, and occurred independently of Cox-1 derived enzymes. In addition, in the mPGES-1 deficient mice, the synthesis of CRH hnRNA and mRNA was decreased in the paraventricular nucleus of the hypothalamus after LPS-challenge, indicating that the delayed hormone secretion was mediated by PGE2-induced gene-transcription of CRH in the hypothalamus. The expression of the c-fos gene and Fos protein, an index of synaptic activation, was maintained in the paraventricular nucleus and its brainstem afferents both after unselective and Cox-2 selective inhibition as well as in Cox-1, Cox-2, and mPGES-1 knock-out mice. This suggests that the immune-induced neuronal activation of autonomic relay nuclei occurs independently of prostanoid synthesis and that it is insufficient for eliciting stress hormone release.

Le développement et l'homéostasie cellulaire de la prostate impliquent le contrôle strict des voies de signalisation induites par les androgènes et les facteurs de croissance. Ces diverses voies sont profondément altérées dans le cancer de la prostate, notamment lors des stades les plus avancés. Dans ce travail, une lignée immortalisée à partir de l'épithélium de prostate humaine, la lignée RWPE-1, a été utilisée pour étudier certains signaux régulant la prolifération cellulaire, ainsi que les connexions entre les voies de signalisation correspondantes. La prolifération des cellules RWPE-1 est sous la dépendance de l'EGF (Epidermal Growth Factor) qui intervient physiologiquement dans le développement épithélial. Les récepteurs apparentés à l'EGF-R sont également impliqués dans la prolifération au cours de la progression tumorale. La prolifération des cellules RWPE-1 en réponse à l'EGF est strictement dépendante de la voie ERK1/2, qui est donc considérée comme un point d'intégration des signaux. L'utilisation d'inhibiteurs du récepteur aux androgènes a permis de montrer le rôle essentiel qu'il joue dans l'activation d'ERK1/2 en réponse à l'EGF. Le récepteur aux androgènes s'associe avec plusieurs molécules de signalisation dans les cellules RWPE-1. Je démontre ici pour la première fois une association entre le récepteur aux androgènes et la kinase Raf-1, activatrice de la voie ERK1/2. Ainsi, le récepteur aux androgènes contrôlerait directement un processus essentiel à la prolifération épithéliale selon un mode d'action non-génomique. Par ailleurs, j'ai montré que la réponse proliférative des cellules RWPE-1 à l'IL-6 requiert l'activation de la voie ERK1/2, et l'activité kinase de l'EGF-R, suggérant la transactivation de ce récepteur par l'IL-6. L'utilisation de divers inhibiteurs chimiques a permis de démontrer que les métalloprotéases de la famille ADAM (a disintegrin and metalloprotease), notamment ADAM17, sont impliquées dans ce processus. Ainsi, l'activation de protéines ADAM par l'IL-6 conduirait au clivage d'un ligand membranaire de l'EGF-R, aboutissant à l'activation de la voie ERK1/2. Ce nouveau mécanisme pourrait être impliqué dans les situations inflammatoires conduisant à une prolifération excessive de l'épithélium prostatique, prélude à la transformation tumorale. En conclusion, les voies de signalisation étudiées sont fortement connectées dans les cellules épithéliales normales. Les deux nouveaux mécanismes décrits ici aboutissent à l'activation des kinases ERK1/2, point d'intégration et de convergence des voies de signalisation dans les cellules épithéliales de prostate normale.

Σκοπός. Η διαβητική κετοξέωση (ΔΚ) και η υπεργλυκαιμική υπερωσμωτική κατάσταση (ΥΥΚ) είναι δύο από τις πιο σοβαρές οξείες επιπλοκές του Σακχαρώδη διαβήτη, που εξακολουθούν να αποτελούν σημαντική αιτία νοσηρότητας και θνητότητας μεταξύ των διαβητικών. Οι λοιμώξεις, συχνός εκλυτικός παράγων και επιπλοκή της ΔΚ και ΥΥΚ, αποτελούν την κύρια αιτία θανάτου και η έγκαιρη διάγνωση και αντιμετώπιση της σήψης είναι κριτικής σημασίας για την επιβίωση των ασθενών. Διερευνήσαμε την επίδραση των ανωτέρω καταστάσεων στην ανοσοποιητική απόκριση, μελετώντας τους υποπληθυσμούς των Τ λεμφοκυττάρων – παραμέτρους οξείας φάσης και την ιντερλευκίνη 6 (IL-6) στο περιφερικό αίμα των ασθενών μας, σε μια προσπάθεια να διαπιστώσουμε τυχόν υποκείμενες διαταραχές και να προσδιορίσουμε πιθανόν διαγνωστικούς και προγνωστικούς δείκτες για τη σήψη. Μέθοδος. Η μελέτη μας περιέλαβε 61 διαβητικούς ασθενείς με ΔΚ ή ΥΥΚ. Ξεχωρίσαμε μια ομάδα ασθενών που είχαν συμπτώματα Συνδρόμου συστηματικής φλεγμονώδους αντίδρασης (SIRS). Προσδιορίσαμε στον ορό όλων των ασθενών τις συγκεντρώσεις των παραγόντων οξείας φάσης (συμπεριλαμβανομένης της C αντιδρώσας πρωτεΐνης ,CRP) και της IL-6 (ως κύρια κυτταροκίνη για την παραγωγή πρωτεϊνών οξείας φάσης), κατά την εισαγωγή και στην ύφεση (μετά τη βελτίωση των συμπτωμάτων και σε κατάσταση ευγλυκαιμίας). Σε μια ομάδα 28 ασθενών με ΔΚ ή ΥΥΚ (σε σύγκριση και με αντίστοιχη ομάδα ελέγχου) μελετήσαμε επιπλέον υποπληθυσμούς των Τ λεμφοκυττάρων, τα ολικά (CD3) / τα βοηθητικά (CD4) / τα κατασταλτικά (CD8) Τ κύτταρα και τα κύτταρα φυσικοί φονείς (ΝΚ) χρησιμοποιώντας μονοκλωνικά αντισώματα και μικροσκόπιο ανοσοφθορισμού, προ και αμέσως μετά τη διόρθωση της μεταβολικής διαταραχής. Αποτελέσματα. Παρατηρήσαμε ότι οι υποπληθυσμοί των Τ λεμφοκυττάρων ήταν σημαντικά ελαττωμένοι κατά την εισαγωγή, συγκρινόμενοι με τους υγιείς μάρτυρες (ενώ οι περισσότερες μελέτες διαβητικών τύπου 1 καταγράφουν αυξημένα βοηθητικά Τ κύτταρα) και παρέμειναν και αμέσως μετά τη διόρθωση της μεταβολικής διαταραχής. Οι ασθενείς που τελικά απεβίωσαν είχαν σημαντικά ελαττωμένους τους υποπληθυσμούς των Τ λεμφοκυττάρων (εκτός των ΝΚ κυττάρων) συγκρινόμενοι και με τους υγιείς μάρτυρες και με όσους ασθενείς επιβίωσαν. Από τους 61 ασθενείς της μελέτης με ΔΚ ή ΥΥΚ, οι 49 ασθενείς είχαν συμπτώματα SIRS. Οι 27 ασθενείς είχαν SIRS χωρίς στοιχεία λοίμωξης, ενώ οι 22 ασθενείς είχαν SIRS με αποδεδειγμένη λοίμωξη. Διαπιστώσαμε σημαντικά αυξημένες συγκεντρώσεις CRP και IL-6 στον ορό των σηπτικών διαβητικών ασθενών συγκριτικά με όσους ασθενείς μας είχαν SIRS χωρίς λοίμωξη. Οι ασθενείς που τελικά απεβίωσαν είχαν σημαντικά πιο αυξημένες συγκεντρώσεις CRP και IL-6 κατά την εισαγωγή, που μειώθηκαν σημαντικά στην ύφεση. Συμπεράσματα. Η διαβητική κετοξέωση και η υπεργλυκαιμική υπερωσμωτική κατάσταση προκαλούν συχνά κλινικό σύνδρομο που ομοιάζει με σύνδρομο συστηματικής φλεγμονώδους αντίδρασης. Διαταραχές στην ισορροπία των υποπληθυσμών των Τ λεμφοκυττάρων, κυρίως η ελάττωση των βοηθητικών Τ κυττάρων μπορεί να συμβάλλουν στην υψηλή θνησιμότητα αυτών των μεταβολικών διαταραχών. Οι μετρήσεις των συγκεντρώσεων C αντιδρώσας πρωτεΐνης και ιντερλευκίνης 6 στον ορό αυτής της ομάδας των διαβητικών ασθενών, είναι ένας χρήσιμος τρόπος αποκλεισμού λοίμωξης και επιβεβαίωσης και παρακολούθησης της σήψης. / Aims / hypothesis. Diabetic ketoacidosis (DKA) and hyperglycemic hyperosmolar state (HHS) are two of the most serious acute complications of diabetes mellitus, being important causes of morbidity and mortality among patients with diabetes. Infection is a common precipitating event in DKA and HHS and the major cause of death. An early diagnosis of sepsis in patients with DKA and HHS is crucial and life saving. We studied the immune responses in these states, investigating the peripheral T lymphocyte subsets, acute phase reactants and interleukin 6 (IL-6) to find out how useful these might be for identifying sepsis. Methods. Sixty one diabetic patients with DKA or HHS were enrolled. Patients with signs and symptoms of systemic inflammatory response syndrome (SIRS) were identified. Acute phase reactants, including serum C-reactive protein (CRP) and IL-6, the main cytokine responsible for the induction of acute phase proteins, were measured (concentrations in peripheral blood) on admission and when patients were clinically improved and were euglycaemic. Peripheral T lymphocyte subsets including total (CD3), helper (CD4) and suppressor (CD8) T cells and natural killer (NK) cells, were studied in twenty one patients with DKA plus seven patients with HHS and twenty eight healthy matched control (using monoclonal antibodies), prior to and after treatment of metabolic disorders. Results. Peripheral T lymphocyte subsets were decreased in the twenty eight patients with DKA and HHS in admission compared to healthy controls (while helper T cells are mostly increased in diabetics type 1), and remained so after treatment of metabolic disorders. Patients who finally died had significantly decreased T lymphocyte subsets (except NK cells) compared with both healthy controls and patients who survived. A total of forty nine out of sixty one patients with DKA and HHS had signs of SIRS. Twenty seven patients had SIRS and no signs of infection and twenty two patients had SIRS due to proven infection. We detected a significant increase in serum CRP and IL-6 values in patients infected compared to patients with no septic SIRS. Patients who finally died had much higher levels of these proteins, while there was a prompt reduction of serum CRP and IL-6 early during remission. Conclusion / interpretation. Diabetic ketoacidosis and hyperglycemic hyperosmolar state can often cause a clinical syndrome resembling systemic inflammatory response syndrome. An imbalance of subpopulations of T lymphocytes, especially decreased helper T cells (CD4), may be correlated with the high morbidity and mortality in these states. Determination of serum C-reactive protein and interleukin-6 is a useful way of early excluding an underlying infection as well as confirming and monitoring sepsis.

Schizophrenia is a mental illness with early adult onset. Prophylactic treatments would be clinically important and therefore we investigated the effect of two interventions: influenza vaccination of pregnant mothers and valproate treatment during late adolescence. Maternal immune response during pregnancy is thought to adversely affect brain development. We sought to assess whether immune activation by influenza vaccine could itself cause behavioural abnormalities in a mouse model. Our data suggest that further work is needed to make firm conclusions about the behavioural effects of the influenza vaccine. The second part of this thesis describes an analysis of valproate treatment on cortical neuron morphology in Disc1 L100P mice, a model for schizophrenia. Valproate was previously shown to prevent the onset of abnormal behaviours in Disc1 L100P mice. Contrary to expectations, valproate decreased apical spine density and the number of dendritic processes rather than reversing the dendritic deficits seen in Disc1 L100P mice.

<p>This study shows that herbs can be effectively screened for potiential bio-activity using in vitro methods. Further studies will be needed to better explore Artemisia afra&rsquo / s effect on immunoregulation, particularly long term effects of the herb on the immune system and its effect on other disease states.</p>

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

Schizophrenia is a mental illness with early adult onset. Prophylactic treatments would be clinically important and therefore we investigated the effect of two interventions: influenza vaccination of pregnant mothers and valproate treatment during late adolescence. Maternal immune response during pregnancy is thought to adversely affect brain development. We sought to assess whether immune activation by influenza vaccine could itself cause behavioural abnormalities in a mouse model. Our data suggest that further work is needed to make firm conclusions about the behavioural effects of the influenza vaccine. The second part of this thesis describes an analysis of valproate treatment on cortical neuron morphology in Disc1 L100P mice, a model for schizophrenia. Valproate was previously shown to prevent the onset of abnormal behaviours in Disc1 L100P mice. Contrary to expectations, valproate decreased apical spine density and the number of dendritic processes rather than reversing the dendritic deficits seen in Disc1 L100P mice.

Objetivos. Avaliar o papel do transplante de células hematopoiéticas (TCH) e da interleucina 6 (IL – 6) sobre vários parâmetros de estresse oxidativo em pacientes com Adrenoleucodistrofia ligada ao cromossomo X (X-ALD). Métodos. A concentração de malondialdeído (MDA), o conteúdo de carbolinas e sulfidrilas e a concentração de ácido hexacosanóico (C26:0) foram quantificados no plasma de pacientes X-ALD antes e após serem submetidos ao TCH. E, a concentração de MDA, a formação de carbonilas e a concentração de IL-6 foram quantificados em plasma e o conteúdo de glutationa reduzida (GSH) foi quantificado em eritrócitos de pacientes X-ALD com fenótipos cerebral infantil (cALD) ou assintomáticos no momento diagnóstico. Resultados. Observamos um aumento significativo na concentração de MDA em plasma de pacientes X-ALD antes e após o TCH em comparação ao grupo controle e uma redução significativa nesses valores após o transplante em comparação aos anteriores ao procedimento. Verificamos uma redução significativa no conteúdo de sulfidrilas no plasma de pacientes X-ALD antes do TCH em comparação ao grupo controle e um aumento significativo desses níveis após o TCH. Não observamos diferenças significativas no conteúdo de carbonilas no plasma de X-ALD antes e após o TCH, em comparação aos controles, apesar de observarmos uma redução significativa nesta determinação nos pacientes após o transplante em relação a antes do TCH. Os pacientes X-ALD apresentam níveis plasmáticos de C26:0 significativamente aumentados antes do TCH em comparação aos controles e, após o TCH, as concentrações de C26:0 foram reduzidas. Observamos uma correlação negativa significativa entre a medida do conteúdo de sulfidrilas e os níveis plasmáticos de C26:0 de indivíduos X-ALD antes do TCH. Também evidenciamos elevados níveis de MDA e da formação de carbonilas no plasma de pacientes cALD e assintomáticos em comparação ao grupo controle. Ainda, observamos redução significativa do conteúdo de GSH nos dois grupos testados comparados aos controles. A quantificação de IL-6 foi significativamente maior nos pacientes cALD, o que não foi observado nos pacientes assintomáticos, apesar destes mostrarem uma tendência de aumento da concentração de IL-6. Conclusões. Os resultados obtidos a partir do plasma de pacientes X-ALD antes e após o TCH demonstram que esta terapia, quando bem indicada e bem sucedida, tem alta efetividade em reduzir a concentração plasmática de C26:0 e é eficaz em reduzir a peroxidação lipídica e o dano oxidativo às proteínas nos pacientes X-ALD. Ainda, é possível relacionar o acúmulo de C26:0 e o dano oxidativo na patogênese da X-ALD. Nossos dados permitem sugerir que a lipoperoxidação e o dano oxidativo às proteínas possam de alguma forma estar envolvidos na fisiopatologia da X-ALD. Além disso, podemos presumir que, nos pacientes X-ALD assintomáticos estudados, o dano oxidativo e os aspectos inflamatórios desempenham papéis importantes na evolução e nas futuras manifestações do fenótipo neuronal. Também podemos supor que a administração de antioxidantes deve ser considerada como uma terapia adjuvante potencial para os pacientes assintomáticos e sintomáticos afetados pela X-ALD, inclusive para aqueles submetidos ao TCH. / Objective. We aimed to evaluate the role of hematopoietic stem cell transplantation (HSCT) and interleukin 6 (IL – 6) on various parameters of oxidative stress in X-linked adrenoleukodystrophy (X-ALD) patients. Methods. Malondialdehyde (MDA), sulfhydryl, carbonyl and hexacosanoic acid (C26:0) levels were measured in plasma from X-ALD patients before and after HSCT. And, MDA, carbonyl and IL-6 levels were measured in plasma and reduced glutathione (GSH) content was measured in erythrocytes from X-ALD patients with different phenotype (asymptomatic and childhood cerebral (CCER patients) at diagnosis moment. Results. We observed increased levels of MDA in plasma from X-ALD before and after HSCT compared to control group, but there was a significant reduction in MDA values after transplantation compared to levels found before the procedure. We verified a significant decrease in sulfhydryl content in plasma of X-ALD patients before HSCT compared with the control group and we also verified a significant increase in the levels of sulfhydryl content after HSCT. No significant differences were observed in carbonyl content in plasma of X-ALD before and after HSCT, compared to controls. However, we observed a significant reduction of plasma carbonyl content from X-ALD patients after HSCT compared to before HSCT. X-ALD patients presented a significant increase of C26:0 plasma level before HSCT when compared to controls and an important reduction of C26:0 plasma concentration in X-ALD patients after HSCT when compared to before HSCT C26:0 levels. We observed an inverse significant correlation between sulfhydryl content and plasma C26:0 levels of X-ALD individuals before HSCT. We also evidenced high levels of MDA and carbonyl formation in plasma from CCER and asymptomatic patients compared to controls. Still, we observed a significant decrease of GSH content in both groups tested compared to controls. The quantification of IL-6 is significantly higher in CCER patients, which is not observed in asymptomatic patients, despite these patients show a tendency of increased concentration of IL-6. Conclusions. The results obtained from plasma of X-ALD patients before and after HSCT demonstrate that this therapy, when well indicated and successful, has high effectiveness in reducing C26:0 plasma and is effective in reducing lipid peroxidation and oxidative damage to proteins in X-ALD patients. Still, it is possible to relate the accumulation of C26:0 and oxidative damage in the pathogenesis of X-ALD. Our data also suggest that lipid peroxidation and protein damage may somehow be involved in the pathophysiology of X-ALD. Moreover, we can assume that in our asymptomatic X-ALD patients, oxidative damage and inflammatory issues seem to play an important role in the evolution and future manifestations of neuronal phenotype. We can also assume that the administration of antioxidants should be considered as a potential adjuvant therapy for asymptomatic and symptomatic patients affected by X-ALD, including those that are submitted to HSCT.

Introduction: Bone is a highly active tissue throughout life and is a subject to constant remodelling. Main cells responsible for continuous resorption and de novo synthesis of bone matrix are osteoclast, osteoblasts and osteocytes. Osteoclasts are the only known type of cells able to resorb bone. These cells are formed by fusion of precursor cells in bone marrow or peripheral blood in a process called osteoclastogenesis. Formation of osteoclasts may be of importance concerning chronic inflammatory diseases that are linked with higher risk of developing osteoporosis during lifespan. Celiac disease is one of those diseases, which is characterized by destruction of intestinal mucosa after ingestion of gluten by susceptible individuals followed by induction of chronic inflammation. In this work, we focused on the potential role of osteoclastogenesis in the development of osteoporosis in patients with celiac disease and we studied roles of selected inflammatory agents (TNF-α, IL-6, IFN-γ a cfDNA) with supposed or hypothesised effects on osteoclastogenesis. Material & Methods: We obtained plasma and serum samples from newly diagnosed patients with celiac disease, patients on gluten free diet and healthy controls and analysed concentrations of cfDNA and inflammatory cytokines TNF-α, IL-6 and IFN-γ in...