Search

Additional file 2: Table S2. Pathway enrichment analysis of PVT1 coexpressed genes.

Additional file 1: Table S1. Core of PVT1 coexpressed genes in adult and pediatric AML, pediatric ALL and DLBCL.

Additional file 1: Fig. S1. Representative box plots relative to IL-23 levels (pg/ml) in plasma from HV (n = 10) and CLL (n = 10). Results are expressed as median and interquartile range. P value shown is obtained from the comparison between the indicated groups by exact non-parametric Mann–Withney U test (*P

This thesis is focused on the key role of extracellular matrix in skin tissue engineering. Firstly, I presented three dimensional structures derived through the physiological secretion of extracellular matrix (ECM), that may be a bioinspired scaffold. I performed a biological characterization of cell-assembled ECMs from three different sub-populations of skin fibroblasts; papillary fibroblasts (Pfi), reticular fibroblasts (Rfi), and dermal papilla fibroblasts (DPfi). Fibroblast sub-populations were cultured with ascorbic acid to promote cell-assembled matrix production for 10 days. Cells were removed and the remaining matrices were characterized. I found that the ECM assembled by Pfi exhibited randomly oriented fibers, associated with highest interfibrillar space, reflecting ECM characteristics which are physiologically present within the papillary dermis. Mass spectrometry followed by immunofluorescence analysis showed that Thrombospondin is preferentially expressed within the DPfi ECM. In another experiment, keratinocytes were seeded on the top of cell depleted ECMs to generate epidermal skin constructs. I found that epidermal constructs grown on DPfi or Pfi matrices exhibited normal basement membrane formation, while Rfi matrices were unable to support membrane formation. Thus, inspiration should be taken from these different ECMs, to design therapeutic biomaterials in skin engineering applications. In the second part, I focused on human decellularized matrix for soft tissue repair and I investigated the biological interaction post-implant of this scaffold. The aims of this second part were to show the clinical results after the application of human decellularized matrix in patients suffering from abdominal hernia and to evaluate the response one year post implant, through morphological analysis of biopsy specimens. Clinical results showed that all the patients revealed a well tolerability of human decellularized matrix. Post-implant morphological results showed cellular repopulation, neo-angiogenesis, minimal inflammatory response and a well-organized collagen matrix in all biopsies. This scaffold can be considered a safe product to treat large abdominal defects.

BACKGROUND; Interest is increasing for human decellularized scaffolds for their ability to favor healing and cell infiltration after transplantation, in the treatment of abdominal wall defects. The purpose of the present study is to show the clinical results obtained after the application of human decellularized dermal matrix (HDM) produced by RER Skin Bank, on patients suffering from different abdominal wall defects. METHODS: Between 2012 and 2014, 64 patients, average age 64 years, received HDM, to replace and cover the damage area during abdominal wall surgery. After surgical procedures, all patients were followed weekly for the first month and then monthly up to 6 months postoperative and any major problem or complication were recorded. Six months follow up included abdominal exams, serological tests and MRI analysis in order to evaluate integration of HDM with the patients surroundings tissues and eventual long-term complications. RESULTS: Incisional hernia was the most frequent clinical condition in which HDM was applied, requiring also the highest amount of human decellularized dermal matrix. One month after the surgical operation, 61 patients revealed a well tolerability of HDM and a normal wound healing was also identified in all the damage areas. Only 3 patients experienced postoperative infections. Moreover the follow up after 6 months reported no signs of dermis rejection and that none of the patients was positive to serological tests. CONCLUSIONS: Human decellularized dermal matrix can be considered a safe and useful bioproduct to treat large abdominal defects, characterized by minor complications and simplicity to be implanted.

Although targeting of cell metabolism is a promising therapeutic strategy in acute myeloid leukemia (AML), metabolic dependencies are largely unexplored. We aimed to classify AML patients based on their metabolic landscape and map connections between metabolic and genomic profiles. Combined serum and urine metabolomics improved AML characterization compared with individual biofluid analysis. At intracellular level, AML displayed dysregulated amino acid, nucleotide, lipid, and bioenergetic metabolism. The integration of intracellular and biofluid metabolomics provided a map of alterations in the metabolism of polyamine, purine, keton bodies and polyunsaturated fatty acids and tricarboxylic acid cycle. The intracellular metabolome distinguished three AML clusters, correlating with distinct genomic profiles: NPM1-mutated(mut), chromatin/spliceosome-mut and TP53-mut/aneuploid AML that were confirmed by biofluid analysis. Interestingly, integrated genomic-metabolic profiles defined two subgroups of NPM1-mut AML. One was enriched for mutations in cohesin/DNA damage-related genes (NPM1/cohesin-mut AML) and showed increased serum choline + trimethylamine-N-oxide and leucine, higher mutation load, transcriptomic signatures of reduced inflammatory status and better ex-vivo response to EGFR and MET inhibition. The transcriptional differences of enzyme-encoding genes between NPM1/cohesin-mut and NPM1-mut allowed in silico modeling of intracellular metabolic perturbations. This approach predicted alterations in NAD and purine metabolism in NPM1/cohesin-mut AML that suggest potential vulnerabilities, worthy of being therapeutically explored.

Sterilization of equipment and tissues is a common clinical practice: there are different chemical, mechanical, and electromagnetic aseptic techniques for inactivating microorganisms. In particular, skin tissue banks are investigating new methods to efficiently decolonize skin tissues, while preserving their structural features. In recent years, cold atmospheric plasma (CAP) has demonstrated bactericidal, virucidal, and fungicidal properties, due to the generation of reactive species and charged particles. For this reason, the aim of this paper is to demonstrate that the implementation of a dielectric barrier discharge (DBD) treatment in air can effectively decontaminate skin tissue from Staphylococcus aureus , retaining cell viability and skin integrity. Fresh skin samples, taken from multitissue donors, were contaminated with Staphylococcus aureus and treated with a DBD source, to verify the level of bacterial decontamination induced by plasma. Cell viability and structural properties of skin tissue were investigated using MTT assay and hematoxylin-eosin staining, respectively. Our results show that CAP can sterilize skin tissue with a bacterial load up to 103 CFU/cm2; moreover, it does not affect cell viability, and no loss of skin structural properties was observed. Thus, CAP treatment could be considered an innovative method for decolonization of human skin, without inducing any microscopic tissue damage, while keeping good cell viability.

The incidence of hepatocellular carcinoma deriving from metabolic dysfunctions has increased in the last years. Sirtuin- (SIRT-3), phospho-mammalian target of rapamycin (p-mTOR) and hypoxia-inducible factor- (HIF-1&alpha, ) are involved in metabolism and cancer. However, their role in hepatocellular carcinoma (HCC) metabolism, drug resistance and progression remains unclear. This study aimed to better clarify the biological and clinical function of these markers in HCC patients, in relation to the presence of metabolic alterations, metformin therapy and clinical outcome. A total of 70 HCC patients were enrolled: 48 and 22 of whom were in early stage and advanced stage, respectively. The expression levels of the three markers were assessed by immunohistochemistry and summarized using descriptive statistics. SIRT-3 expression was higher in diabetic than non-diabetic patients, and in metformin-treated than insulin-treated patients. Interestingly, p-mTOR was higher in patients with metabolic syndrome than those with different etiology, and, similar to SIRT-3, in metformin-treated than insulin-treated patients. Moreover, our results describe a slight, albeit not significant, benefit of high SIRT-3 and a significant benefit of high nuclear HIF-1&alpha, expression in early-stage patients, whereas high levels of p-mTOR correlated with worse prognosis in advanced-stage patients. Our study highlighted the involvement of SIRT-3 and p-mTOR in metabolic dysfunctions that occur in HCC patients, and suggested SIRT-3 and HIF-1&alpha, as predictors of prognosis in early-stage HCC patients, and p-mTOR as target for the treatment of advanced-stage HCC.

Background In chronic lymphocytic leukemia (CLL) disease onset and progression are influenced by the behavior of specific CD4+ T cell subsets, such as T regulatory cells (Tregs). Here, we focused on the phenotypic and functional characterization of Tregs in CLL patients to improve our understanding of the putative mechanism by which these cells combine immunosuppressive and effector-like properties. Methods Peripheral blood mononuclear cells were isolated from newly diagnosed CLL patients (n = 25) and healthy volunteers (n = 25). The phenotypic and functional characterization of Tregs and their subsets was assessed by flow cytometry. In vitro analysis of TH1, TH2, TH17 and Tregs cytokines was evaluated by IFN-γ, IL-4, IL-17A and IL-10 secretion assays. The transcriptional profiling of 84 genes panel was evaluated by RT2 Profiler PCR Array. Statistical analysis was carried out using exact non parametric Mann–Whitney U test. Results In all CLL samples, we found a significant increase in the frequency of IL-10-secreting Tregs and Tregs subsets, a significant rise of TH2 IL-4+ and TH17 IL-17A+ cells, and a higher percentage of IFN-γ/IL-10 and IL-4/IL-10 double-releasing CD4+ T cells. In addition, we also observed the up-regulation of innate immunity genes and the down-regulation of adaptive immunity ones. Conclusions Our data show that Tregs switch towards an effector-like phenotype in CLL patients. This multifaceted behavior is accompanied by an altered cytokine profiling and transcriptional program of immune genes, leading to a dysfunction in immune response in the peripheral blood environment of CLL patients. Electronic supplementary material The online version of this article (10.1186/s12967-018-1545-0) contains supplementary material, which is available to authorized users.