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La ricerca verte sulla comprensione del ruolo del ferro e della ferroptosi in diversi contesti patologici. La ferroptosi è una forma di morte cellulare non apoptotica, ferro-dipendente, causata dai ROS. Ci siamo focalizzati sulla leucemia linfoblastica acuta di tipo B (LLA-B), uno dei tumori ematologici più frequenti nei bambini. Per avere una prima idea sul ruolo del ferro LLA-B, si è valutata l'espressione di geni ferro in campioni di midollo osseo. Si è lavorato principalmente su modelli cellulari di LLA-B, i quali rappresentano diversi sottotipi di LLA. Le cellule sono esposte a concentrazioni crescenti di ferro: a dosi basse il ferro promuove la proliferazione cellulare, a livelli elevati si dimostra citotossico. È interessante notare che solamente una delle quattro linee cellulari mostra una risposta favorevole ad alte concentrazioni di ferro. Questa linea cellulare, RS4;11, è caratterizzata dalla traslocazione t(4;11), tipica della leucemia a linea cellulare mista (MLL). MLL è associata a un'insorgenza improvvisa, è molto aggressiva e a una prognosi sfavorevole. Utilizzando diversi approcci sperimentali, si sono studiate le risposte cellulari post-somministrazione di ferro e RSL3, induttore di ferroptosi. I risultati dimostrano che le cellule dipendono fortemente dal ferro per la loro crescita. Inoltre, si è dimostrato che RSL3 innesca perossidazione lipidica, produzione di ROS e morte cellulare. Tutti questi processi vengono bloccati in presenza di Liproxstatin-1, un inibitore di ferroptosi. Confermato che i modelli in vitro mostrano risposte diverse all'induzione della ferroptosi, si è ipotizzato che questa diversa sensibilità possa essere attribuita all'attivazione di diversi set di geni che conferiscono resistenza all'induzione della ferroptosi. La comprensione del ruolo del metabolismo del ferro e dell'impatto della ferroptosi in varie patologie rappresenta il fulcro del progetto di dottorato. Studiando il metabolismo del ferro nel LLA-B, il g, The research was focused on understanding the role of iron and ferroptosis in different pathological contexts. Ferroptosis is a form of non-apoptotic, iron-dependent cell death caused by lipid-based reactive oxygen species (ROS). Firstly, we focused on B-acute lymphoblastic leukaemia (B-ALL), the most diagnosed cancer in children. To obtain an initial understanding of iron's role in B-ALL, we assessed the expression of iron-related genes in patients’ bone marrow samples. Then, we worked on four B-ALL in vitro models, which represented different B-ALL subtypes. We exposed cells to varying iron concentrations, observing that lower doses promoted cell proliferation, while higher levels proved cytotoxic. Interestingly, one (RS4;11) out of the four cell lines exhibited a favourable response to high iron concentrations. RS4;11 is characterized by translocation t(4;11), which is typical of MLL (Mixed-Lineage Leukaemia) – rearranged leukaemia. MLL-r leukaemia is associated with a sudden onset, aggressive progression, and notoriously poor. Using different experimental approaches, we studied cellular responses in two MLL cell lines after administrations of iron and RSL3, ferroptosis inducer. Our results demonstrated that cell lines strongly depend on iron for growth and expansion. Moreover, we showed that the RSL3 triggers lipid peroxidation, production of ROS and cell death. All these processes are arrested in the presence of Liproxstatin-1, a small-molecule inhibitor that prevents lipid peroxidation. Moreover, we confirmed that in vitro MLL models exhibited distinct responses to ferroptosis induction. We postulated that this varying sensitivity might be attributed to the activation of different gene sets capable of conferring resistance to ferroptosis. Understanding the role of iron metabolism and the impact of ferroptosis in various diseases settings represented the core of the PhD project. While studying iron metabolism in B-ALL, Ceruloplasmin protein resulted as one of t

Nowadays, the increase of the unconventional oil deposit exploitation and the amount of oil sands process-affected waters (OSPW) in tailing ponds emerges the importance of developing bio-monitoring strategies for the restoration of these habitats. The major constituents of such deposits are naphthenic acids (NAs), emerging contaminant mixtures with toxic and recalcitrant properties. With the aim of developing bio-monitoring strategies based on culture-independent approach, we identified genes coding for enzymes involved in NA degradation from Rhodococcus opacus R7 genome, after the evaluation of its ability to mineralize model NAs. R. opacus R7 whole-genome analysis unveiled the presence of pobA and chcpca gene clusters putatively involved in NAs degradation. Gene expression analysis demonstrated the specific induction of R7 aliA1 gene, encoding for a long-chain-fatty-acid-CoA ligase, in the presence of cyclohexanecarboxylic acid (CHCA) and hexanoic acid (HA), selected as representative compounds for alicyclic and linear NAs, respectively. Therefore, aliA1 gene was selected as a molecular marker to monitor the biodegradative potential of slurry-phase sand microcosms in different conditions: spiked with CHCA, in the presence of R. opacus R7, the autochthonous microbial community, and combining these factors. Results revealed that the aliA1-targeting culture-independent approach could be a useful method for bio-monitoring of NA degradation in a model laboratory system.

Background. Several inherited diseases cause hyperferritinemia with or without iron overload. Differential diagnosis is complex and requires an extensive work-up. Currently, a clinical-guided approach to genetic tests is performed based on gene-by-gene sequencing. Although rea-sonable, this approach is expensive and time-consuming and Next Generation Sequencing (NGS) technology may provide cheaper and quicker large-scale DNA sequencing. Methods. We analysed 36 patients with non-HFE-related hyperferritinemia. Liver iron concentration was measured in 33 by magnetic resonance. A panel of 25 iron related genes was designed using SureDesign software. Custom libraries were generated and then sequenced using Ion Torrent PGM. Results. We identified six novel mutations in SLC40A1, three novel and one known mutation in TFR2, one known mutation and a de-novo deletion in HJV, and a novel mutation in HAMP in ten patients. In silico analyses supported the pathogenic role of the mutations. Conclusions. Our results support the use of an NGS-based panel in selected patients with hyperferritinemia in a tertiary center for iron metabolism disorders. However, 26 out of 36 patients did not show genetic variants that can individually explain hyperferritinemia and/or iron overload suggesting the existence of other genetic defects or gene-gene and gene-environment interactions needing further studies.

Ferroptosis is an iron-dependent form of cell death characterized by intracellular lipid peroxide accumulation and redox imbalance. Ferroptosis shows specific biological and morphological features when compared to the other cell death patterns. The loss of lipid peroxide repair activity by glutathione peroxidase 4 (GPX4), the presence of redox-active iron and the oxidation of polyunsaturated fatty acid (PUFA)-containing phospholipids are considered as distinct fingerprints of ferroptosis. Several pathways, including amino acid and iron metabolism, ferritinophagy, cell adhesion, p53, Keap1/Nrf2 and phospholipid biosynthesis, can modify susceptibility to ferroptosis. Through the decades, various diseases, including acute kidney injury; cancer; ischemia-reperfusion injury; and cardiovascular, neurodegenerative and hepatic disorders, have been associated with ferroptosis. In this review, we provide a comprehensive analysis of the main biological and biochemical mechanisms of ferroptosis and an overview of chemicals used as inducers and inhibitors. Then, we report the contribution of ferroptosis to the spectrum of liver diseases, acute or chronic. Finally, we discuss the use of ferroptosis as a therapeutic approach against hepatocellular carcinoma, the most common form of primary liver cancer.

Introduction: Adolescents with cancer often experience a longer diagnostic delay than children, mainly because they take longer to go to a doctor. The Italian Society for Adolescents with Oncohematological Diseases (SIAMO) has launched an information campaign focusing on raising adolescents' awareness of the importance of diagnosing cancer early. Methods: The concepts of the campaign were developed by a scientific committee of clinicians, cancer patients and their parents, and marketing experts. The title of the campaign is "There's no reason why". A video has been launched on TV channels and the Internet, and the final frame refers viewers to the SIAMO website, which provides advice to help adolescents interpret any symptoms they experience. Results: The video has had 12,181 views. In the 6 months following the launch of the campaign, the SIAMO website page dedicated to the campaign was opened by 9,767 viewers for a total of 13,632 views. Conclusions: Though it remains very difficult to judge the efficacy of this initiative, the value of a campaign focusing on improving the adolescent population's cancer awareness is supported by the large number of studies published on the diagnostic delay in this age group. Our campaign goes to show the importance of ensuring cooperation between the different stakeholders involved in the global care of adolescents with cancer.

Non-small cell lung cancer (NSCLC) harboring chromosomal rearrangements of the anaplastic lymphoma kinase (ALK) gene is treated with ALK tyrosine kinase inhibitors (TKI), but the treatment is successful for only a limited amount of time; most patients experience a relapse due to the development of drug resistance. Here, we show that a vaccine against ALK induced a strong and specific immune response that both prophylactically and therapeutically impaired the growth of ALK-positive lung tumors in mousemodels. The ALK vaccine was efficacious also in combination with ALK TKI treatment and significantly delayed tumor relapses after TKI suspension. We found that lung tumors containing ALK rearrangements induced an immunosuppressive microenvironment, regulating the expression of PD-L1 on the surface of lung tumor cells. High PD-L1 expression reduced ALK vaccine efficacy, which could be restored by administration of anti- PD-1 immunotherapy. Thus, combinations of ALK vaccine with TKIs and immune checkpoint blockade therapies might represent a powerful strategy for the treatment of ALK-driven NSCLC.

The aim of this trial was to assess the effects of the administration of different combinations of mycotoxins in naturally contaminated maize grains on dairy heifer growth, blood measurements and puberty onset. A total of 35 Friesian female heifers were randomly allotted to three experimental groups from 18-21 to 42-45 weeks of age. During the 24-week experimental period (EP), heifers were fed the same diet, but with maize meal derived from three differently contaminated lots: very low contamination, as control (C); medium-low aflatoxin-contaminated (A); and mixed aflatoxin-fumonisin contaminated (A-F). At the end of the EP, they returned to a common diet without contaminated maize, and they were monitored for an additional period of 12 weeks (post-experimental period, PEP). BW, wither height, hip height, body length and heart girth were measured every 4 weeks from the beginning of EP to the end of PEP. At the same time, body condition score was evaluated and blood samples were taken from the jugular vein to be analysed for haematological, serum protein and metabolic profiles. Age at puberty was assessed by measuring weekly plasma progesterone levels from 40 to 52 weeks of age. Body growth measurements were processed both by ANOVA of average daily gain of EP and PEP separately, and by the analysis of growth curve parameters. Haematological, serum protein and metabolic profile were evaluated using a mixed model, taking into account the repeated measurements in time on each animal. Heifers' growth was delayed both in A and A-F groups during EP, as evidenced by the different linear coefficients of the BW growth curve in the three groups. Differently contaminated diets did not affect the haematological profile, so that it can be concluded that these levels of mycotoxin contamination do not determine any specific effect on haematopoiesis and immunity in growing heifers. The main blood marker of mycotoxin chronic toxicity was the.-glutamyl transferase activity level in pl

The aim of this paper was to compare body growth, hematological profile development, and clinical biochemistry in the female progeny of a sire with the female progeny of its clone. Sixteen Friesian female calves, 9 daughters from a tested bull (BULL) and 7 from its somatic cell nuclear transfer clone (CLONE) were monitored from birth to 60 wk of life. Body weight (BW), wither height (WH), hip height (HH), body length (BL), and hearth girth (HG) were measured at birth and 4, 8, 12, 16, 20, 24, 36, and 50 wk. Blood samples were taken from jugular vein at 12 to 48 h from birth and 1, 2, 3, 4, 8, 12, 16, 20, 24, and 36 wks of age, to be analyzed for hematological, serum protein, and metabolic profiles. At the same time, rectal temperature (RT) was recorded. Age at puberty was assessed on surviving heifers by measuring weekly plasma progesterone levels. Data were evaluated using a mixed model, taking into account the repeated measures in time on the calf. For each variable, different covariance structures were tested, choosing the best according to the Akaike’s Information Criteria. Significant was set at P 0.05, and a trend was considered for P 0.10. At 24 wk of age,WHwas lower in CLONE daughters than BULL daughters. Around 20 wk of age, there was a trend for lower BW in CLONE daughters than BULL daughters, confirmed from differences in HG. There was no difference in RT due to sire effect. Blood glucose concentration decreased in both groups during the first 4 wk of life; at birth, only a trend for higher blood glucose in CLONE daughters was recorded, whereas an opposite trend was observed for plasma creatinine. Total leukocyte count did not differ between progenies. Circulating lymphocytes tended to be lower in CLONE than BULL daughters. The neutrophil: lymphocyte ratio tended to be higher in CLONE than BULL calves. No difference was demonstrated for erythrocyte features, whereas mean platelet volume tended to be lower in CLONE than BULL progeny. From these results, th