Maximizing the antioxidant capacity of Padina pavonica by choosing the right drying and extraction methods

Marine algae are becoming an interesting source of biologically active compounds with a promising application as nutraceuticals, functional food ingredients, and therapeutic agents. The effect of drying (freeze-drying, oven-drying, and shade-drying) and extraction methods (shaking at room temperature, shaking in an incubator at 60 \ub0C, ultrasound-assisted extraction (UAE), and microwave-assisted extraction (MAE)) on the total phenolics content (TPC), total flavonoids content (TFC), and total tannins content (TTC), as well as antioxidant capacity of the water/ethanol extracts from Padina pavonica were investigated. The TPC, TFC, and TTC values of P. pavonica were in the range from 0.44 \ub1 0.03 to 4.32 \ub1 0.15 gallic acid equivalents in mg/g (mg GAE/g) dry algae, from 0.31 \ub1 0.01 to 2.87 \ub1 0.01 mg QE/g dry algae, and from 0.32 \ub1 0.02 to 10.41 \ub1 0.62 mg CE/g dry algae, respectively. The highest TPC was found in the freeze-dried sample in 50% ethanol, extracted by MAE (200 W, 60 \ub0C, and 5 min). In all cases, freeze-dried samples extracted with ethanol (both 50% and 70%) had the higher antioxidant activity, while MAE as a green option reduces the extraction time without the loss of antioxidant activity in P. pavonica

PRIN LEVANTE 2020: Levulinic acid valorization through advanced novel technologies

The project LEVANTE deals with the development of new catalytic processes for the valorization of levulinic acid and its esters towards three classes of compounds: levulinic ketals, diphenolic acid and γ-valerolactone together with other reduction products

Blastic plasmacytoid dendritic cell neoplasm : State of the art and prospects

Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is an extremely rare tumour, which usually a_ects elderly males and presents in the skin with frequent involvement of the bone-marrow, peripheral blood and lymph nodes. It has a dismal prognosis, with most patients dying within one year when treated by conventional chemotherapies. The diagnosis is challenging, since neoplastic cells can resemble lymphoblasts or small immunoblasts, and require the use of a large panel of antibodies, including those against CD4, CD56, CD123, CD303, TCL1, and TCF4. The morphologic and in part phenotypic ambiguity explains the uncertainties as to the histogenesis of the neoplasm that led to the use of various denominations. Recently, a series of molecular studies based on karyotyping, gene expression profiling, and next generation sequencing, have largely unveiled the pathobiology of the tumour and proposed the potentially beneficial use of new drugs. The latter include SL-401, anti-CD123 immunotherapies, venetoclax, BET-inhibitors, and demethylating agents. The epidemiologic, clinical, diagnostic, molecular, and therapeutic features of BPDCN are thoroughly revised in order to contribute to an up-to-date approach to this tumour that has remained an orphan disease for too long

Predictive and Prognostic Molecular Factors in Diffuse Large B-Cell Lymphomas

Diffuse large B-cell lymphoma (DLBCL) is the commonest form of lymphoid malignancy, with a prevalence of about 40% worldwide. Its classification encompasses a common form, also termed as "not otherwise specified" (NOS), and a series of variants, which are rare and at least in part related to viral agents. Over the last two decades, DLBCL-NOS, which accounts for more than 80% of the neoplasms included in the DLBCL chapter, has been the object of an increasing number of molecular studies which have led to the identification of prognostic/predictive factors that are increasingly entering daily practice. In this review, the main achievements obtained by gene expression profiling (with respect to both neoplastic cells and the microenvironment) and next-generation sequencing will be discussed and compared. Only the amalgamation of molecular attributes will lead to the achievement of the long-term goal of using tailored therapies and possibly chemotherapy-free protocols capable of curing most (if not all) patients with minimal or no toxic effects

Blastic plasmacytoid dendritic cell neoplasm: Genomics mark epigenetic dysregulation as a primary therapeutic target

Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare and aggressive hematologic malignancy for which there is still no effective B therapy. In order to identify genetic alterations useful for a new treatment design, we used whole-exome sequencing to analyze 14 BPDCN patients and the patient-derived CAL-1 cell line. The functional enrichment analysis of mutational data reported the epigenetic regulatory program to be the most significantly undermined (P<0.0001). In particular, twenty-five epigenetic modifiers were found mutated (e.g. ASXL1, TET2, SUZ12, ARID1A, PHF2, CHD8); ASXL1 was the most frequently affected (28.6% of cases). To evaluate the impact of the identified epigenetic mutations at the gene-expression and Histone H3 lysine 27 trimethylation/acetylation levels, we performed additional RNA and pathology tissue-chromatin immunoprecipitation sequencing experiments. The patients displayed enrichment in gene signatures regulated by methylation and modifiable by decitabine administration, shared common H3K27-acetylated regions, and had a set of cell-cycle genes aberrantly up-regulated and marked by promoter acetylation. Collectively, the integration of sequencing data showed the potential of a therapy based on epigenetic agents. Through the adoption of a preclinical BPDCN mouse model, established by CAL-1 cell line xenografting, we demonstrated the efficacy of the combination of the epigenetic drugs 5’-azacytidine and decitabine in controlling disease progression in vivo

PO-272 Leukemia-associated NPM mutations promote quiescence of hematopoietic stem cells and prevent their functional exhaustion upon oncogene-induced hyper-proliferation

Introduction Acute Myeloid Leukaemia (AML) is a heterogeneous and multi-step disease. The serial acquisition of mutations and the environmental pressure allow one or more clones to expand and contribute to the disease. In particular, 6% of AMLs are characterised by an initial mutation in the DNMT3a gene, followed by mutations in NPM (NPMc) and FLT3 loci (FLT3-ITD). We previously shown that NPMc can drive AML development in mouse model and highly cooperates with FLT3-ITD. Moreover, it has been reported that normal Hematopoietic Stem Cells (HSCs) of elderly people may bear some somatic early AML mutations and this correlate with an increased risk of hematologic diseases suggesting that mutations can shape pre-leukemic HSCs to be more prone to the acquisition of further mutations giving rise to Leukaemia Initiating Cells (LIC). While the ability of FLT3-ITD to drive HSC compartment exhaustion has been already shown, the impact of NPMc on HSCs remains unclear. Material and methods Taking advantage of the extended pre-leukemic phase of our inducible NPMc mouse model, we elucidate the role of NPMc in HSCs by functional and transcriptional analysis. Moreover, to investigate the basis of NPMc and FLT3-ITD cooperation we generate mice carrying both the conditional NPMc transgene and the FLT3-ITD constitutive mutation and, before AML onset, we analyse double mutant HSCs behaviour. Results and discussions We have found that NPMc expression lead to the expansion of the HSC compartment through the enforcement of a stem-cell transcriptional program that increases self-renewal by promoting quiescence. We then investigated how the NPMc dependent quiescence program is linked to its oncogenic function. The expression of NPMc +in the FLT3-ITD background prevents the HSCs exhaustion imposed by FLT3-ITD and restores their repopulating capacity. Accordingly, gene expression analysis revealed a strong dominance of NPMc +with the restoration of the same transcriptional program observed in the NPMc HSCs. These data strongly suggest that NPMc imposes a HSC-specific program that, in combination with the oncogenic signal provided by FLT3-ITD, allows the selection of the LIC and the occurrence of AML. Conclusion In conclusion, enforcement of quiescence might be a critical function for the maintenance of the transformed clone during both the pre-leukemic and the leukemic phase. As consequence, interfering with quiescence key determinants may eradicate the reservoir of quiescent cells responsible for disease recurrence

Pathogenetic and diagnostic significance of microRNA deregulation in peripheral T-cell lymphoma not otherwise specified

Peripheral T-cell lymphomas not otherwise specified (PTCLs/NOS) are rare and aggressive tumours whose molecular pathogenesis and diagnosis are still challenging. The microRNA (miRNA) profile of 23 PTCLs/NOS was generated and compared with that of normal T-lymphocytes (CD4+, CD8+, naive, activated). The differentially expressed miRNA signature was compared with the gene expression profile (GEP) of the same neoplasms. The obtained gene patterns were tested in an independent cohort of PTCLs/NOS. The miRNA profile of PTCLs/NOS then was compared with that of 10 angioimmunoblastic T-cell lymphomas (AITLs), 6 anaplastic large-cell lymphomas (ALCLs)/ALK+ and 6 ALCLs/ALK - . Differentially expressed miRNAs were validated in an independent set of 20 PTCLs/NOS, 20 AITLs, 19 ALCLs/ALK - and 15 ALCLs/ALK+. Two hundred and thirty-six miRNAs were found to differentiate PTCLs/NOS from activated T-lymphocytes. To assess which miRNAs impacted on GEP, a multistep analysis was performed, which identified all miRNAs inversely correlated to different potential target genes. One of the most discriminant miRNAs was selected and its expression was found to affect the global GEP of the tumours. Moreover, two sets of miRNAs were identified distinguishing PTCL/NOS from AITL and ALCL/ALK - , respectively. The diagnostic accuracy of this tool was very high (83.54%) and its prognostic value validated

Dual targeting of the DNA damage response pathway and BCL-2 in diffuse large B-cell lymphoma

Standard chemotherapies for diffuse large B-cell lymphoma (DLBCL), based on the induction of exogenous DNA damage and oxidative stress, are often less effective in the presence of increased MYC and BCL-2 levels, especially in the case of double hit (DH) lymphomas harboring rearrangements of the MYC and BCL-2 oncogenes, which enrich for a patient’s population characterized by refractoriness to anthracycline-based chemotherapy. Here we hypothesized that adaptive mechanisms to MYC-induced replicative and oxidative stress, consisting in DNA damage response (DDR) activation and BCL-2 overexpression, could represent the biologic basis of the poor prognosis and chemoresistance observed in MYC/BCL-2-positive lymphoma. We first integrated targeted gene expression profiling (T-GEP), fluorescence in situ hybridization (FISH) analysis, and characterization of replicative and oxidative stress biomarkers in two independent DLBCL cohorts. The presence of oxidative DNA damage biomarkers identified a poor prognosis double expresser (DE)-DLBCL subset, characterized by relatively higher BCL-2 gene expression levels and enrichment for DH lymphomas. Based on these findings, we tested therapeutic strategies based on combined DDR and BCL-2 inhibition, confirming efficacy and synergistic interactions in in vitro and in vivo DH-DLBCL models. These data provide the rationale for precision-therapy strategies based on combined DDR and BCL-2 inhibition in DH or DE-DLBCL

NR1H3 (LXRα) is associated with pro-inflammatory macrophages, predicts survival and suggests potential therapeutic rationales in diffuse large b-cell lymphoma

The role of macrophages (Mo) and their prognostic impact in diffuse large B-cell lymphomas (DLBCL) remain controversial. By regulating the lipid metabolism, Liver-X-Receptors (LXRs) control Mo polarization/inflammatory response, and their pharmacological modulation is under clinical investigation to treat human cancers, including lymphomas. Herein, we surveyed the role of LXRs in DLBCL for prognostic purposes. Comparing bulk tumors with purified malignant and normal B-cells, we found an intriguing association of NR1H3, encoding for the LXR-α isoform, with the tumor microenvironment (TME). CIBERSORTx-based purification on large DLBCL datasets revealed a high expression of the receptor transcript in M1-like pro-inflammatory Mo. By determining an expression cut-off of NR1H3, we used digital measurement to validate its prognostic capacity on two large independent on-trial and real-world cohorts. Independently of classical prognosticators, NR1H3high patients displayed longer survival compared with NR1H3low cases and a high-resolution Mo GEP dissection suggested a remarkable transcriptional divergence between subgroups. Overall, our findings indicate NR1H3 as a Mo-related biomarker identifying patients at higher risk and prompt future preclinical studies investigating its mouldability for therapeutic purposes

Blastic plasmacytoid dendritic cell neoplasm: genomics mark epigenetic dysregulation as a primary therapeutic target

Blastic Plasmacytoid Dendritic Cell Neoplasm is a rare and aggressive hematological malignancy currently lacking an effective therapy. To possibly identify genetic alterations useful for a new treatment design, we analyzed by whole-exome sequencing fourteen Blastic Plasmacytoid Dendritic Cell Neoplasm patients and the patient-derived CAL-1 cell line. The functional enrichment analysis of mutational data reported the epigenetic regulatory program as the most significantly undermined (P<.0001). In particular, twenty-five epigenetic-modifiers were found mutated (e.g., ASXL1, TET2, SUZ12, ARID1A, PHF2, CHD8); ASXL1 was the most frequently affected (28.6% of cases). To evaluate the impact of the identified epigenetic mutations at the gene-expression and Histone H3 lysine 27 trimethylation/acetylation levels, we performed additional RNA and Pathology tissue-chromatin immunoprecipitation sequencing experiments; the patients displayed enrichment in gene-signatures regulated by methylation and modifiable by Decitabine administration, shared common H3K27-acetylated regions and featured a set of cell-cycle genes aberrantly up-regulated and marked by promoter acetylation. Collectively, the integration of sequencing data showed the potential of a therapy based on epigenetic agents. Through the adoption of a preclinical Blastic Plasmacytoid Dendritic Cell Neoplasm mouse model, established by the CAL-1 cell line xenografting, we demonstrated the efficacy of the combination of the epigenetic drugs 5'-Azacytidine and Decitabine in controlling the disease progression in vivo