Parasites of cage cultured European seabass <i>Dicentrarchus Labrax</i> and gilthead seabream <i>Sparus aurata</i> from Sardinia (western Mediterranean): first results

European seabass Dicentrarchus labrax and gilthead seabream Sparus aurata are the most important marine finfish species intensively cultured in the Mediterranean. Many factors influenced the rapid increase in the production of these species in the last two decades. One of the most important factors is the great development and diffusion of sea-cage culture, because some of the parasite species has become a serious threat to cage-reared fish in other Mediterranean localities

Effect of two different protein/fat ratios of the diet on meagre (<i>Argyrosomus regius</i>) traits

The aim of this study was to evaluate the effect of two diets with different protein/fat ratios (P/F) (diet A: P/F 2.26; diet B: P/F 3.36) on the chemical composition, fatty acid profile and some somatic indexes of meagre (Argyrosomus regius). The trial was carried out on two groups of meagre raised in two different sea cages during 15 months. At the end of the production cycle biometric measures as well as chemical-nutritional analysis of the fillets were conducted on 25 fishes per group. Diet A, with a lower P/F, furnished animals with higher percentages of mesenteric fat (0.48 vs 0.41%; P&lt;0.01) and of fillet yield (51.21 vs 48.12; P&lt;0.01). Moreover, the fillets obtained with the diet A showed higher percentage of fat (3.60 vs 2.41%; P&lt;0.01), lower moisture (74.10 vs 75.42%; P&lt;0.01), lower losses of water under pressure (16.73 vs 20.20%; P&lt;0.01) and after 48 h of refrigeration (3.08 vs 4.23%; P&lt;0.01). The fatty acids profile of fillets was affected by the diet. Diet A resulted in a higher level of saturated fatty acids (26.44 vs 23.17% of total lipid; P&lt;0.01) and a lower percentage of polyunsaturated fatty acids (31.56 vs 36.08%; P&lt;0.01) in the fillet, mainly due to the lower content of linoleic acid (13.63 vs 19.77%; P&lt;0.01). The atherogenic (AI) and thrombogenic (TI) indexes, which resulted very low in the fish of Group B (AI=0.48 vs 0.60, P&lt;0.01; TI=0.33 vs 0.37, P&lt;0.01), together with the low lipid content of meat in both groups, confirmed the very high nutritional quality of meagre fillets

Effect of two different protein/fat ratios of the diet on meagre (Argyrosomus regius) traits

The aim of this study was to evaluate the effect of two diets with different protein/fat ratios (P/F) (diet A: P/F 2.26; diet B: P/F 3.36) on the chemical composition, fatty acid profile and some somatic indexes of meagre (Argyrosomus regius). The trial was carried out on two groups of meagre raised in two different sea cages during 15 months. At the end of the production cycle biometric measures as well as chemical-nutritional analysis of the fillets were conducted on 25 fishes per group. Diet A, with a lower P/F, furnished animals with higher percentages of mesenteric fat (0.48 vs 0.41%; P<0.01) and of fillet yield (51.21 vs 48.12; P<0.01). Moreover, the fillets obtained with the diet A showed higher percentage of fat (3.60 vs 2.41%; P<0.01), lower moisture (74.10 vs 75.42%; P<0.01), lower losses of water under pressure (16.73 vs 20.20%; P<0.01) and after 48 h of refrigeration (3.08 vs 4.23%; P<0.01). The fatty acids profile of fillets was affected by the diet. Diet A resulted in a higher level of saturated fatty acids (26.44 vs 23.17% of total lipid; P<0.01) and a lower percentage of polyunsaturated fatty acids (31.56 vs 36.08%; P<0.01) in the fillet, mainly due to the lower content of linoleic acid (13.63 vs 19.77%; P<0.01). The atherogenic (AI) and thrombogenic (TI) indexes, which resulted very low in the fish of Group B (AI=0.48 vs 0.60, P<0.01; TI=0.33 vs 0.37, P<0.01), together with the low lipid content of meat in both groups, confirmed the very high nutritional quality of meagre fillets

Valproic acid triggers erythro/megakaryocyte lineage decision through induction of GFI1B and MLLT3 expression

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miR-494-3p overexpression promotes megakaryocytopoiesis in primary myelofibrosis hematopoietic stem/progenitor cells by targeting SOCS6

Primary myelofibrosis (PMF) is a chronic Philadelphia-negative myeloproliferative neoplasm characterized by hematopoietic stem cell-derived clonal myeloproliferation, involving especially the megakaryocyte lineage. To better characterize how the altered expression of microRNAs might contribute to PMF pathogenesis, we have previously performed the integrative analysis of gene and microRNA expression profiles of PMF hematopoietic stem/progenitor cells (HSPCs), which allowed us to identify miR- 494-3p as the upregulated microRNA predicted to target the highest number of downregulated mRNAs. To elucidate the role of miR-494-3p in hematopoietic differentiation, in the present study we demonstrated that miR-494-3p enforced expression in normal HSPCs promotes megakaryocytopoiesis. Gene expression profiling upon miR-494-3p overexpression allowed the identification of genes commonly downregulated both after microRNA overexpression and in PMF CD34+ cells. Among them, suppressor of cytokine signaling 6 (SOCS6) was confirmed to be a miR-494-3p target by luciferase assay. Western blot analysis showed reduced level of SOCS6 protein as well as STAT3 activation in miR-494-3p overexpressing cells. Furthermore, transient inhibition of SOCS6 expression in HSPCs demonstrated that SOCS6 silencing stimulates megakaryocytopoiesis, mimicking the phenotypic effects observed upon miR-494-3p overexpression. Finally, to disclose the contribution of miR-494-3p upregulation to PMF pathogenesis, we performed inhibition experiments in PMF HSPCs, which showed that miR-494-3p silencing led to SOCS6 upregulation and impaired megakaryocyte differentiation. Taken together, our results describe for the first time the role of miR-494- 3p during normal HSPC differentiation and suggest that its increased expression, and the subsequent downregulation of its target SOCS6, might contribute to the megakaryocyte hyperplasia commonly observed in PMF patients

An automated 3D-printed perfusion bioreactor combinable with pulsed electromagnetic field stimulators for bone tissue investigations

In bone tissue engineering research, bioreactors designed for replicating the main features of the complex native environment represent powerful investigation tools. Moreover, when equipped with automation, their use allows reducing user intervention and dependence, increasing reproducibility and the overall quality of the culture process. In this study, an automated uni-/bi-directional perfusion bioreactor combinable with pulsed electromagnetic field (PEMF) stimulation for culturing 3D bone tissue models is proposed. A user-friendly control unit automates the perfusion, minimizing the user dependency. Computational fluid dynamics simulations supported the culture chamber design and allowed the estimation of the shear stress values within the construct. Electromagnetic field simulations demonstrated that, in case of combination with a PEMF stimulator, the construct can be exposed to uniform magnetic fields. Preliminary biological tests on 3D bone tissue models showed that perfusion promotes the release of the early differentiation marker alkaline phosphatase. The histological analysis confirmed that perfusion favors cells to deposit more extracellular matrix (ECM) with respect to the static culture and revealed that bi-directional perfusion better promotes ECM deposition across the construct with respect to uni-directional perfusion. Lastly, the Real-time PCR results of 3D bone tissue models cultured under bi-directional perfusion without and with PEMF stimulation revealed that the only perfusion induced a ~ 40-fold up-regulation of the expression of the osteogenic gene collagen type I with respect to the static control, while a ~ 80-fold up-regulation was measured when perfusion was combined with PEMF stimulation, indicating a positive synergic pro-osteogenic effect of combined physical stimulation

Development and Multicenter Validation of a Novel Immune-Inflammation-Based Nomogram to Predict Survival in Western Resectable Gastric and Gastroesophageal Junction Adenocarcinoma (GEA): The NOMOGAST

Background. More than 50% of operable GEA relapse after curative-intent resection. We aimed at externally validating a nomogram to enable a more accurate estimate of individualized risk in resected GEA. Methods. Medical records of a training cohort (TC) and a validation cohort (VC) of patients undergoing radical surgery for c/uT2-T4 and/or node-positive GEA were retrieved, and potentially interesting variables were collected. Cox proportional hazards in univariate and multivariate regressions were used to assess the effects of the prognostic factors on OS. A graphical nomogram was constructed using R software’s package Regression Modeling Strategies (ver. 5.0-1). The performance of the prognostic model was evaluated and validated. Results. The TC and VC consisted of 185 and 151 patients. ECOG:PS > 0 (p < 0.001), angioinvasion (p < 0.001), log (Neutrophil/Lymphocyte ratio) (p < 0.001), and nodal status (p = 0.016) were independent prognostic values in the TC. They were used for the construction of a nomogram estimating 3- and 5-year OS. The discriminatory ability of the model was evaluated with the c-Harrell index. A 3-tier scoring system was developed through a linear predictor grouped by 25 and 75 percentiles, strengthening the model’s good discrimination (p < 0.001). A calibration plot demonstrated a concordance between the predicted and actual survival in the TC and VC. A decision curve analysis was plotted that depicted the nomogram’s clinical utility. Conclusions. We externally validated a prognostic nomogram to predict OS in a joint independent cohort of resectable GEA; the NOMOGAST could represent a valuable tool in assisting decision-making. This tool incorporates readily available and inexpensive patient and disease characteristics as well as immune-inflammatory determinants. It is accurate, generalizable, and clinically effectivex

Role of TGF-\u3b21/miR-382-5p/SOD2 axis in the induction of oxidative stress in CD34+ cells from primary myelofibrosis

Primary myelofibrosis (PMF) is a myeloproliferative neoplasm characterized by an excessive production of pro-inflammatory cytokines resulting in chronic inflammation and genomic instability. Besides the driver mutations in JAK2, MPL, and CALR genes, the deregulation of miRNA expression may also contribute to the pathogenesis of PMF. To this end, we recently reported the upregulation of miR-382-5p in PMF CD34+ cells. In order to unveil the mechanistic details of the role of miR-382-5p in pathogenesis of PMF, we performed gene expression profiling of CD34+ cells overexpressing miR-382-5p. Among the downregulated genes, we identified superoxide dismutase 2 (SOD2), which is a predicted target of miR-382-5p. Subsequently, we confirmed miR-382-5p/SOD2 interaction by luciferase assay and we showed that miR-382-5p overexpression in CD34+ cells causes the decrease in SOD2 activity leading to reactive oxygen species (ROS) accumulation and oxidative DNA damage. In addition, our data indicate that inhibition of miR-382-5p in PMF CD34+ cells restores SOD2 function, induces ROS disposal, and reduces DNA oxidation. Since the pro-inflammatory cytokine transforming growth factor-\u3b21 (TGF-\u3b21) is a key player in PMF pathogenesis, we further investigated the effect of TGF-\u3b21 on ROS and miR-382-5p levels. Our data showed that TGF-\u3b21 treatment enhances miR-382-5p expression and reduces SOD2 activity leading to ROS accumulation. Finally, inhibition of TGF-\u3b21 signaling in PMF CD34+ cells by galunisertib significantly reduced miR-382-5p expression and ROS accumulation and restored SOD2 activity. As a whole, this study reports that TGF-\u3b21/miR-382-5p/SOD2 axis deregulation in PMF cells is linked to ROS overproduction that may contribute to enhanced oxidative stress and inflammation. Our results suggest that galunisertib may represent an effective drug reducing abnormal oxidative stress induced by TGF-\u3b21 in PMF patients. Database linking: GEO: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE103464

miRNA-mRNA integrative analysis in primary myelofibrosis CD34+ cells: role of miR-155/JARID2 axis in abnormal megakaryopoiesis

Primary myelofibrosis (PMF) is a myeloproliferative neoplasm characterized by megakaryocyte (MK) hyperplasia, bone marrow fibrosis, and abnormal stem cell trafficking. PMF may be associated with somatic mutations in JAK2, MPL, or CALR. Previous studies have shown that abnormal MKs play a central role in the pathophysiology of PMF. In this work, we studied both gene and microRNA (miRNA) expression profiles in CD34(+) cells from PMF patients. We identified several biomarkers and putative molecular targets such as FGR, LCN2, and OLFM4. By means of miRNA-gene expression integrative analysis, we found different regulatory networks involved in the dysregulation of transcriptional control and chromatin remodeling. In particular, we identified a network gathering several miRNAs with oncogenic potential (eg, miR-155-5p) and targeted genes whose abnormal function has been previously associated with myeloid neoplasms, including JARID2, NR4A3, CDC42, and HMGB3. Because the validation of miRNA-target interactions unveiled JARID2/miR-155-5p as the strongest relationship in the network, we studied the function of this axis in normal and PMF CD34(+) cells. We showed that JARID2 downregulation mediated by miR-155-5p overexpression leads to increased in vitro formation of CD41(+) MK precursors. These findings suggest that overexpression of miR-155-5p and the resulting downregulation of JARID2 may contribute to MK hyperplasia in PMF

Annali storici di Principato Citra, A. 7, n. 1.1 (2009)

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