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In the present study, the effect of Rosmarinus officinalis L. dietary supplementation on meat quality and oxidative stability of Nero Siciliano pigs was examined. During the growing–fattening period, 32 Nero Siciliano pigs were allotted into two treatment groups consisting of 8 replicates with 2 pigs per pen. For 90 days, the animals received a basal diet: one group (CTR) was not dietary supplemented, whereas the other group received (1 g/kg) rosemary extract (ROX). Supplementation with rosemary extract significantly improved the polyunsaturated fatty acid content of the meat, which showed higher values in the meat of the ROX group compared with the CTR group (P0.05). Color measurement performed in the present study on meat samples from the two dietary treatments showed that redness decreased (P=0.046) and hue values increased (P=0.036), indicating that a deterioration of the initial color occurred and that the rosemary extract was ineffective in preventing color deterioration. Nevertheless, the lightness, yellowness and chroma color descriptors showed similar values in relation to dietary treatment (P>0.05). Considering the nutritional value of meat as an important contributor to the overall quality, the results obtained in this study support the possibility of the dietary supplementation with R. officinalis L. extract in pigs as a functional additive in livestock feeding.

During the growing-fattening period (93 days; ILW 33.5±6 kg to FLW 67±3 kg), 30 Nero Siciliano pigs were fed on a basal diet supplemented with (ROX group) or not (CTR group) a rosemary extract (1g?kg-1). At 1, 3 and 5 days after slaughtering, the oxidative stability of the Longissimus dorsi muscle was determined by using TBArs test. Moreover, on the individual samples of the muscle the lipid content, the fatty acid and the sterol composition were determined; the acidic composition of the lard, removed from backfat, was also studied. Data were subjected to ANOVA. The fat content of the muscle was unaffected by the dietary treatment as well as the oxidative stability whereas, significant differences (P²0.01) were observed for the polyunsaturated fatty acid content which showed the highest values in the meat as well as in the lard of the ROX group; PUFA/SFA ratio was also significantly highest in the muscle (P=0.004) and in the lard (P=0.017) of the ROX group, testifying a possible antioxidative activity of the rosemary extract. The sterol fraction (cholesterol, cholestanol, stigmasterol, beta-sitosterol and delta 5-avenasterol) of the Longissimus dorsi muscle was unaffected by the rosemary supplementation; a significant difference was observed only for the campesterol (CTR group=1.08, ROX group=0.90; P = 0.021).

Background: Hematopoietic cell transplantation (HCT) is an effective treatment for pediatric patients with high-risk, refractory, or relapsed acute myeloid leukemia (AML). However, a large proportion of transplanted patients eventually die due to relapse. To improve overall survival, we propose a combined strategy based on cord blood (CB)-HCT with the application of AML-specific T cell receptor (TCR)-engineered T cell therapy derived from the same CB graft., Methods: We produced CB-CD8 + T cells expressing a recombinant TCR (rTCR) against Wilms tumor 1 (WT1) while lacking endogenous TCR (eTCR) expression to avoid mispairing and competition. CRISPR-Cas9 multiplexing was used to target the constant region of the endogenous TCRα ( TRAC ) and TCRβ ( TRBC ) chains. Next, an optimized method for lentiviral transduction was used to introduce recombinant WT1-TCR. The cytotoxic and migration capacity of the product was evaluated in coculture assays for both cell lines and primary pediatric AML blasts., Results: The gene editing and transduction procedures achieved high efficiency, with up to 95% of cells lacking eTCR and over 70% of T cells expressing rWT1-TCR. WT1-TCR-engineered T cells lacking the expression of their eTCR (eTCR -/- WT1-TCR) showed increased cell surface expression of the rTCR and production of cytotoxic cytokines, such as granzyme A and B, perforin, interferon-γ (IFNγ), and tumor necrosis factor-α (TNFα), on antigen recognition when compared with WT1-TCR-engineered T cells still expressing their eTCR (eTCR +/+ WT1-TCR). CRISPR-Cas9 editing did not affect immunophenotypic characteristics or T cell activation and did not induce increased expression of inhibitory molecules. eTCR -/- WT1-TCR CD8 + CB-T cells showed effective migratory and killing capacity in cocultures with neoplastic cell lines and primary AML blasts, but did not show toxicity toward healthy cells., Conclusions: In summary, we show the feasibility of developing a potent CB-derived CD8 + T cell product targeting WT1, providing an option for post-transplant allogeneic immune cell therapy or as an off-the-shelf product, to prevent relapse and improve the clinical outcome of children with AML., Competing Interests: Competing interests: JK reports grants from Gadeta, Novartis, and Miltenyi Biotec and is the inventor of patents dealing with γδT cell-related aspects, as well as the cofounder and shareholder of Gadeta. ZS is an inventor of patents dealing with γδT cell-related aspects., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Faced with the national emergency linked to the spread of COVID-19 in Italy, digital technologies have made it possible to carry out the ordinary activities of the various educational agencies through the main tool of Distance Learning (DaD). The intensive use of information and communication technologies (ICT) and the guarantee of accessibility represent an enabling prerogative for current education systems, enriching training in a variety of ways and opportunities which must be accessible to all. ICTs take on a propulsive function for change in schools because they make it possible to affect the school setting, to transform the learning environment by redesigning space, reorganizing time, modifying communication and socialization processes, encouraging, in students, the development of key competencies in digital literacy and media education. Considering the context of a school transformed and renewed by the teaching and training potential of ICT, it becomes central to reconstruct the requests and needs developed by the practitioners of educational policies to cope with the reorganization of teaching methods and times at the time of DaD. Starting from these premises, the paper focuses attention on the social impact (Stern, 2016) of DaD to evaluate: the extent and intensity of the methodological-didactic innovation required of teachers for the organization and conducting remote lessons; the increase-in students-of transversal and digital literacy skills (team working, problem solving, etc.) potentially associated with the use of ICTs; the involvement and collaboration of families in the process of assessing and verifying learning. The reflection is part of a broader research project by the University of Sapienza University of Rome entitled "The social impact assessment of DaD after COVID-19"; a 3 year evaluation research addressed to a typological sample of upper secondary schools in Rome classified on the basis of the Infrastructure and Equipment indicator of the Rav and the social effect of the school (school effect) on the academic performance of students in the tests Invalsi. The evaluation aimed to identify-from the DaD experience-indications useful in re-designing the school's intervention strategy in the phases following the pandemic; for this reason it adopted an analysis perspective that valorized the positive and most successful aspects in the testimonies of the teachers and students involved in the first phase of the research (conducted in May-June 2021). Within the framework of the Positive Thinking Evaluation, the empirical evidence-collected through the administration/conduction of semi-structured interviews, focus groups, online ethnographic observation of the lessons in DaD-will allow us to reflect on some dimensions of success and of particular social innovation for the teachers' teaching practices and the students' learning processes in DaD. In the Positive Thinking Evaluation, success is a positive effect (not just a "good practice"), even an unexpected one, of an activity that has produced a positive change in the context of program implementation., Competing Interests: The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer EC declared a past collaboration with the author to the handling editor., (Copyright © 2023 Lo Presti.)

Background: Immunotherapy in high-risk neuroblastoma (HR-NBL) does not live up to its full potential due to inadequate (adaptive) immune engagement caused by the extensive immunomodulatory capacity of HR-NBL. We aimed to tackle one of the most notable immunomodulatory processes in neuroblastoma (NBL), absence of major histocompatibility complex class I (MHC-I) surface expression, a process greatly limiting cytotoxic T cell engagement. We and others have previously shown that MHC-I expression can be induced by cytokine-driven immune modulation. Here, we aimed to identify tolerable pharmacological repurposing strategies to upregulate MHC-I expression and therewith enhance T cell immunogenicity in NBL., Methods: Drug repurposing libraries were screened to identify compounds enhancing MHC-I surface expression in NBL cells using high-throughput flow cytometry analyses optimized for adherent cells. The effect of positive hits was confirmed in a panel of NBL cell lines and patient-derived organoids. Compound-treated NBL cell lines and organoids were cocultured with preferentially expressed antigen of melanoma (PRAME)-reactive tumor-specific T cells and healthy-donor natural killer (NK) cells to determine the in vitro effect on T cell and NK cell cytotoxicity. Additional immunomodulatory effects of histone deacetylase inhibitors (HDACi) were identified by transcriptome and translatome analysis of treated organoids., Results: Drug library screening revealed MHC-I upregulation by inhibitor of apoptosis inhibitor (IAPi)- and HDACi drug classes. The effect of IAPi was limited due to repression of nuclear factor kappa B (NFκB) pathway activity in NBL, while the MHC-I-modulating effect of HDACi was widely translatable to a panel of NBL cell lines and patient-derived organoids. Pretreatment of NBL cells with the HDACi entinostat enhanced the cytotoxic capacity of tumor-specific T cells against NBL in vitro, which coincided with increased expression of additional players regulating T cell cytotoxicity (eg, TAP1/2 and immunoproteasome subunits). Moreover, MICA and MICB, important in NK cell cytotoxicity, were also increased by entinostat exposure. Intriguingly, this increase in immunogenicity was accompanied by a shift toward a more mesenchymal NBL cell lineage., Conclusions: This study indicates the potential of combining (immuno)therapy with HDACi to enhance both T cell-driven and NKcell-driven immune responses in patients with HR-NBL., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Immunotherapies targeting checkpoint inhibition and cell therapies are considered breakthroughs for cancer therapy. However, only a part of patients benefit from these treatments and resistance has been observed. Combining both approaches can potentially further enhance their efficacy. With the advent of gene editing techniques, such as clustered regularly interspaced short palindromic repeats-CRISPR associated protein 9 (CRISPR-Cas9), the elimination of checkpoint molecules became available as an option in good manufacturing practice conditions to improve persistence and efficacy. However, no data of CRISPR-Cas9 application have been reported in cord blood (CB)-derived immune cells, potentially usable for allogeneic cell therapy purposes. In this article, we describe the optimization of a protocol to deplete checkpoint molecules at the genomic level using CRISPR-Cas9 technology from CB-dendritic cells (DCs) and CB-CD8 + T cells. The protocol is based on the electroporation of a ribonucleoprotein complex, easily translatable to clinical settings. In both cell types, the knock-out (KO) was successful and did not affect cell viability. CB-DCs showed a decrease in expression of the targeted protein ranging from 50% to 95%, while CB-CD8 + T cells showed a reduction in the range of 25-45%. The procedure did not affect the stimulatory function of the CB-DCs or the response of CB-CD8 + T cells (proliferation or TNF-α production). In conclusion, we optimized a protocol to eliminate checkpoint molecules from CB-derived DCs and CD8 + T cells, with the aim to further implement allogeneic cell therapies for cancer.

The chemical and microbiological characteristics of filets of Spaurus aurata L. specimens fed with diets containing a Hermetia illucens meal (HIM) at the 25, 35, and 50%, as a partial replacement for fish meal (FM) were evaluated. The diets, formulated to satisfy the nutritional needs of fish, were isoenergetic (22 MJ/kg gross energy), isonitrogenous (43 g/100 g, a.f.), and isolipidic (19 g/100 g, a.f.). Seventy-two specimens were randomly killed after 186 days of growing trials. Then, the filets were analyzed for chemical profile, fatty acids, amino acids, minerals, and microbial flora. Data were subjected to statistical analysis. No significant differences were observed in chemical composition. The sum of polyunsaturated fatty acids (PUFAs) showed a similar content in the filets; eicosapentaenoic acid was similar in the filets of HIM0, HIM35%, and HIM50%, whereas docosahexaenoic acid was higher in filets of the HIM0 group. n3/n6 PUFA ratio and the sum of EPA + DHA showed a high value ( p < 0.001) in filets of the group fed with FM. No significant difference was observed in thrombogenic index and hypocholesterolaemic/hypercholesterolaemic ratio in the groups; the atherogenic index showed a higher value ( p = 0.001) in the HIM50% group. Indispensable amino acids showed some significant ( p < 0.0001) differences in the groups; arginine and phenylalanine content was higher in the filets of fish fed with FM; isoleucine and valine content was higher in the filets of HIM50%; leucine, lysine and methionine content was lower in the filets of HIM35%; histidine content was lower in the filets of HIM25%; tryptophan content was lower in filets of the HIM50% group. EAA/NEAA ratio showed highest value in the filets of the group that received FM. The presence of HIM in the three diets kept chromium, manganese, iron, copper, zinc, and nickel levels lower than those recommended by various authorities. Ca/P ratio showed a higher level ( p < 0.0001) in the group fed with FM than those fed with diets containing HIM. The insect meal in the diets did not influence the microbiological profile of fish. Use of HIM as an unconventional feed ingredient in Sparus aurata diet looks promising, although the quality of filets may be affected., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Oteri, Chiofalo, Maricchiolo, Toscano, Nalbone, Lo Presti and Di Rosa.)

Objective: This systematic review and meta-analysis aimed to summarise the available evidence on the pre- and intra-operative risk factors for anastomotic leakage (AL) after bowel resection and anastomosis for ovarian cancer (OC)., Study Design: We searched online databases from Pubmed, Scopus, ScienceDirect, and Cochrane Library from inception to October 2020. Pre- and intra-operative risk factors for AL were considered as the primary outcomes. Research heterogeneity and bias were evaluated by I 2 and by the Newcastle Ottawa scale, respectively. The study was registered with PROSPERO, CRD42018095225., Results: The overall AL rate after OC surgery (median ± SD) was 5.3 ± 12% (277 AL on 5178 anastomoses). Thirteen non-randomised studies were included in the meta-analysis enrolling a total of 3274 patients. Pre albumin level ≤ 3 gr/dl, multiple bowel resections and primary cytoreductive surgery were associated with a significantly high risk of AL with a pooled OR of 5.29 (95% CI: 1.51-18.59), OR = 4.4 (95% CI: 1.19-16.66) and OR = 1.71 (95% CI: 1.05-2.77), respectively. Optimal cytoreduction, ASA score, ascites, and protective stoma were not associated with an increased risk of AL., Conclusion: Based on the best available evidence, preoperative albumin level <3 gr/dl, multiple bowel resections and primary cytoreductive surgery were associated with an increased risk for AL after bowel surgery for OC., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021. Published by Elsevier B.V.)

The determination of alkaline phosphatase (ALP) in cheeses has become an official method for controlling cheeses with a protected designation of origin (PDO), all of which use raw milk. PDO cheeses, characterized by high craftsmanship, usually have an uneven quality. However, for these cheeses, it is necessary to establish ALP values so that they can be defined as a raw milk product. In this study, a dataset with Pecorino Siciliano PDO samples was analyzed to determine ALP both at the core and under the rind. The results showed that there was no significant difference between the different zones in Pecorino cheese. A second dataset of 100 pecorino cheese samples determined that ALP was only at the core of the cheese. Moreover, there was a statistically significant difference between the ALP values of cheeses produced with raw milk and those produced with pasteurized milk. Furthermore, according to the temperatures, a wide variability of ALP values was observed in the Pecorino Siciliano PDO samples from the core of the cheeses. This was a result of several under scotta whey cooking methodologies adopted by cheesemakers, which do not permit a clear range. Therefore, further investigation is desirable.

Competing Interests: Competing interests: None declared.

With the aim to explore the use of A. hypochondriacus seeds for animal feeding, the agronomic traits, nutrients, and bioactive compounds of four accessions with different origin (India, Nebraska, Iowa, and Pennsylvania) grown in a Mediterranean environment were studied. Proximate composition was determined using the official methods of analyses, fatty acid profile by gas chromatography, total phenolic content (TPC) and the scavenging activity (DPPH • and ABTS •+ ) by colorimetric method. A one-way ANOVA model was performed to determine the differences between accessions. The four A. hypochondriacus accessions showed interesting seed yield results. No significant differences were observed for crude protein and crude fiber; the oil content showed the significant highest values in the seeds from Nebraska and Pennsylvania, but their nutritional characteristics were significantly different. The accession from Nebraska showed the highest oleic and linoleic acid levels, the highest values of polyunsaturated fatty acids, the best atherogenic and thrombogenic indices and hypocholesterolemic/hypercholesterolaemic ratio, and the highest TPC content. The accession from Pennsylvania showed the highest antioxidant activity and lowest peroxidation index. On the whole, A. hypochondriacus seeds can be used as pseudo-cereal to balance the animal diet and the accession should be chosen according to the different metabolic pathways of unsaturated fatty acids in ruminant and monogastric animals.

Adoptive T cell therapy utilizing tumor-specific autologous T cells has shown promising results for cancer treatment. However, the limited numbers of autologous tumor-associated antigen (TAA)-specific T cells and the functional aberrancies, due to disease progression or treatment, remain factors that may significantly limit the success of the therapy. The use of allogeneic T cells, such as umbilical cord blood (CB) derived, overcomes these issues but requires gene modification to induce a robust and specific anti-tumor effect. CB T cells are readily available in CB banks and show low toxicity, high proliferation rates, and increased anti-leukemic effect upon transfer. However, the combination of anti-tumor gene modification and preservation of advantageous immunological traits of CB T cells represent major challenges for the harmonized production of T cell therapy products. In this manuscript, we optimized a protocol for expansion and lentiviral vector (LV) transduction of CB CD8 + T cells, achieving a transduction efficiency up to 83%. Timing of LV treatment, selection of culture media, and the use of different promoters were optimized in the transduction protocol. LentiBOOST was confirmed as a non-toxic transduction enhancer of CB CD8 + T cells, with minor effects on the proliferation capacity and cell viability of the T cells. Positively, the use of LentiBOOST does not affect the functionality of the cells, in the context of tumor cell recognition. Finally, CB CD8 + T cells were more amenable to LV transduction than peripheral blood (PB) CD8+ T cells and maintained a more naive phenotype. In conclusion, we show an efficient method to genetically modify CB CD8 + T cells using LV, which is especially useful for off-the-shelf adoptive cell therapy products for cancer treatment., Competing Interests: N.P.v.T. is an employee of AVROBIO, Inc., Cambridge, MA, USA. J.K. is cofounder and shareholder of Gadeta; is an inventor on multiple patents dealing with γδTCR topics as well as isolation of engineered immune cells; and received research support from Miltenyi Biotec, Novartis, and Gadeta. The other authors declare no competing interests., (© 2021 The Authors.)

Recent developments in gene engineering technologies have drastically improved the therapeutic treatment options for cancer patients. The use of effective chimeric antigen receptor T (CAR-T) cells and recombinant T cell receptor engineered T (rTCR-T) cells has entered the clinic for treatment of hematological malignancies with promising results. However, further fine-tuning, to improve functionality and safety, is necessary to apply these strategies for the treatment of solid tumors. The immunosuppressive microenvironment, the surrounding stroma, and the tumor heterogeneity often results in poor T cell reactivity, functionality, and a diminished infiltration rates, hampering the efficacy of the treatment. The focus of this review is on recent advances in rTCR-T cell therapy, to improve both functionality and safety, for potential treatment of solid tumors and provides an overview of ongoing clinical trials. Besides selection of the appropriate tumor associated antigen, efficient delivery of an optimized recombinant TCR transgene into the T cells, in combination with gene editing techniques eliminating the endogenous TCR expression and disrupting specific inhibitory pathways could improve adoptively transferred T cells. Armoring the rTCR-T cells with specific cytokines and/or chemokines and their receptors, or targeting the tumor stroma, can increase the infiltration rate of the immune cells within the solid tumors. On the other hand, clinical "off-tumor/on-target" toxicities are still a major potential risk and can lead to severe adverse events. Incorporation of safety switches in rTCR-T cells can guarantee additional safety. Recent clinical trials provide encouraging data and emphasize the relevance of gene therapy and gene editing tools for potential treatment of solid tumors.

Essential oils (EOs) obtained from aromatic plants are rich in natural components with interesting antimicrobial effects. The aim of this study was to evaluate the chemical composition of EOs extracted from Origanum majorana (OM-EO), Mentha suaveolens (MS-EO), Rosmarinus officinalis (RO-EO), Salvia officinalis (SO-EO) and Mentha pulegium (MP-EO). Their antioxidant properties and antibacterial activity against Listeria monocytogenes and different serotypes of Salmonella enterica subsp. enterica were also studied. The EOs were extracted from plants by hydro-distillation and their chemical composition was determined by GC-MS. Terpinen-4-ol, 1,8-Cineole, Camphor, Limonene and Cinerone were the main chemical components found in OM-EO, RO-EO, SO-EO, MP-EO and MS-EO, respectively. To the best of our knowledge, Limonene and Cinerone were reported, for the first time, as the major components of MP-EO and MS-EO. Moreover, our results showed that MS-EO had the best antioxidant activity with an IC 50 of 0.78 ± 0.05 mg/mL, EC 50 of 1.53 ± 0.07 mg/mL, and RC 50 of 0.98 ± 0.04 mg/mL, and the higher antibacterial activity using microdilution broth method with MIC of 0.5% for Salmonella and 0.25% for L. monocytogenes, while OM-EO had the best antibacterial activity using disc diffusion method (inhibition diameters ranged between 15.3 ± 0.3 mm and 18.5 ± 0.3 mm for Salmonella and between 20.1 ± 0.2 mm and 25.4 ± 0.4 mm for L. monocytogenes). However, OM-EO and MS-EO present the higher percentage of sub-lethally injured cells against S. enterica (5.50 ± 0.11%) and L. monocytogenes (5.23 ± 0.07%), respectively. From this study, we can conclude that the investigated EOs are rich in components with interesting antibacterial activity and they could be applied in food preparations as natural preservatives to extend the shelf life of food products and to inhibit the growth of food-borne pathogens., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

β-Thalassemia intermedia is a disorder characterized by ineffective erythropoiesis (IE), anemia, splenomegaly, and systemic iron overload. Novel approaches are being explored based on the modulation of pathways that reduce iron absorption (ie, using hepcidin activators like Tmprss6-antisense oligonucleotides [ASOs]) or increase erythropoiesis (by erythropoietin [EPO] administration or modulating the ability of transferrin receptor 2 [Tfr2] to control red blood cell [RBC] synthesis). Targeting Tmprss6 messenger RNA by Tmprss6-ASO was proven to be effective in improving IE and splenomegaly by inducing iron restriction. However, we postulated that combinatorial strategies might be superior to single therapies. Here, we combined Tmprss6-ASO with EPO administration or removal of a single Tfr2 allele in the bone marrow of animals affected by β-thalassemia intermedia (Hbbth3/+). EPO administration alone or removal of a single Tfr2 allele increased hemoglobin levels and RBCs. However, EPO or Tfr2 single-allele deletion alone, respectively, exacerbated or did not improve splenomegaly in β-thalassemic mice. To overcome this issue, we postulated that some level of iron restriction (by targeting Tmprss6) would improve splenomegaly while preserving the beneficial effects on RBC production mediated by EPO or Tfr2 deletion. While administration of Tmprss6-ASO alone improved the anemia, the combination of Tmprss6-ASO + EPO or Tmprss6-ASO + Tfr2 single-allele deletion produced significantly higher hemoglobin levels and reduced splenomegaly. In conclusion, our results clearly indicate that these combinatorial approaches are superior to single treatments in ameliorating IE and anemia in β-thalassemia and could provide guidance to translate some of these approaches into viable therapies., (© 2020 by The American Society of Hematology.)

Hematopoietic cell transplantation (HCT) is a last resort, potentially curative treatment option for pediatric patients with refractory acute myeloid leukemia (AML). Cord blood transplantation (CBT) results in less relapses and less graft-versus-host disease when compared to other sources. Nevertheless, still more than half of the children die from relapses. We therefore designed a strategy to prevent relapses by inducing anti-AML immunity after CBT, using a CB-derived dendritic cell (CBDC) vaccine generated from CD34+ CB cells from the same graft. We here describe the optimization and validation of good manufacturing practice (GMP)-grade production of the CBDC vaccine. We show the feasibility of expanding low amounts of CD34+ cells in a closed bag system to sufficient DCs per patient for at least three rounds of vaccinations. The CBDCs showed upregulated costimulatory molecules after maturation and showed enhanced CCR7-dependent migration toward CCL19 in a trans-well migrations assay. CBDCs expressed Wilms' tumor 1 (WT1) protein after electroporation with WT1 -mRNA, but were not as potent as CBDCs loaded with synthetic long peptides (peptivator). The WT1-peptivator loaded CBDCs were able to stimulate T-cells both in a mixed lymphocyte reaction as well as in an antigen-specific (autologous) setting. The autologous stimulated T-cells lysed not only the WT1+ cell line, but most importantly, also primary pediatric AML cells. Altogether, we provide a GMP-protocol of a highly mature CBDC vaccine, loaded with WT1 peptivator and able to stimulate autologous T-cells in an antigen-specific manner. Finally, these T-cells lysed primary pediatric AML demonstrating the competence of the CBDC vaccine strategy., (Copyright © 2020 Plantinga, Lo Presti, de Haar, Dünnebach, Madrigal, Lindemans, Boelens and Nierkens.)

Agronomic traits, oil content, fatty acid composition, antioxidant activity, and total phenolic content were studied on eight A. cruentus accessions cultivated in Southern Italy. A one-way ANOVA model was performed to compare accessions and the Principal Components Analysis was applied to identify patterns in our dataset and highlight similarities and differences. A. cruentus showed valuable seed yield (0.27 kg/m 2 , on average) comparable to the main tradition cereals used for animal feeding. Seed-oil composition showed significant differences among the accessions. Data showed a higher lipid content than most cereal grains (from 5.6 to 7.3%). Approximately 60% of fatty acids were unsaturated; linoleic fatty acid ranged from 19 to 34%, oleic acid from 29 to 36%, and alfa-linolenic fatty acid from 0.3 to 0.5%, respectively. The saturated/unsaturated fatty acid ratio ranged from 0.5 to 0.8, the hypocholesterolemic:hypercholesterolaemic ratio from 1.7 to 2.7, the Atherogenic Index from 0.38 to 0.66, the Thrombogenic Index from 0.85 to 1.48, the total phenolic content from 0.14 to 0.36 mg/g seeds, and the antioxidant activity (DPPH • ) from 0.30 to 0.50. The studied seed-oil composition evidenced A. cruentus as a healthy ingredient for animal feed and consequently, as a possible substitute for traditional cereals. Accessions from Mexico and Arizona emerged for their high qualitative traits.

Dietary partially destoned olive cake supplementation on performance, carcass traits and meat quality of intensively finished bulls was evaluated. Forty-five Limousin bulls, divided into three homogenous groups, received a diet with no supplementation (Control-CTR), 7.5% (Low Olive Cake-LOC), and 15.0% of olive cake supplementation (High Olive Cake-HOC). The trial was realized for 150 days; all bulls were individually weighed at the beginning, middle, and end of the trial, to calculate the individual average daily gain (ADG). At slaughtering, on each carcass, hot weight was recorded and, after 7 days, the pH and temperature were measured. On Longissimus lumborum muscle, color, cooking loss, and shear force of the cooked sample were determined. The chemical composition and the fatty acid content of muscle were determined. Olive cake inclusions (7.5% and 15.0%) increased ( p < 0.05) the body weight, ADG, slaughter traits and intramuscular fat content and influenced ( p < 0.05) the quality indices. The 15.0% of the inclusion reduced ( p < 0.05) the cooking loss and shear force, and increased the unsaturated fatty acid content. The olive cake can be considered as a functional component in beef production and, in substitution to a quote of cereals into the diet of bulls, could be an opportunity to improve agriculture sustainability.

Aim of the present study was to assess the effect of dietary dried partially destoned olive cake supplement on nutritional quality and sensory profile of milk and cheese produced by dairy cows. The experiment was carried out on eighty-four healthy dairy Friesian cows divided into two homogenous groups. The control group (CTR) received a conventional diet, whereas the experimental group (OC) received a conventional diet supplemented with olive cake as 15% of DM. The trial lasted five months. Monthly, on individual milk samples, yield and physical-chemical parameters were determined. Milk was used for the artisanal cheese production. On 10 samples of cheese for each group, physical-chemical and fatty acid profile were determined. Electronic nose analysis and sensory evaluation were performed. Data were analyzed by ANOVA. The diet affected ( p < 0.05) the milk yield, exclusively in September. Yield and quality of cheese of OC group after 60 d of ripening showed higher ( p < 0.05) yield, moisture and fat content, lower ( p < 0.05) pH, protein, salt and ash content, higher ( p < 0.01) MUFA and PUFA and CLA content, lower ( p < 0.05) SFA, higher ( p < 0.01) UFA/SFA and hypocholesterolemic/hypercholesterolemic ratios, better ( p < 0.01) atherogenic and thrombogenic indices. Data show dietary olive cake supplementation in lactating dairy cows improves nutritional and nutraceutical properties of cheese, volatile profile and level of assessors' acceptance.

In poultry production, the intensive use of high-performing hybrid animals led to loss of genetic variability and a consequent lower response to climatic change and disease. Poultry biodiversity is seriously threatened, and its safeguard is a strong objective in developed countries. According to the FAO, which emphasized the importance of native breeds for its country of origin, the aim of this study was to present the first contribution on eggs quality for endangered the Siciliana chicken breed and deepen knowledge on the local Livorno breed. At 20 weeks of age, 108 laying hens (54 Siciliana breed and 54 Livorno breed) were divided into six homogeneous groups of 18 hens each and reared according to requirements imposed by the EC Regulation 889/08 for organic production. The production cycle was controlled over one year, and egg production was recorded daily by group. Eggs were collected, weighted, and measured. Physico-chemical parameter and fatty acids profile were analyzed and nutritional indexes calculated. The statistical model included the effects of breed (Siciliana, Livorno). Egg production was 190 egg/head for Siciliana and 180 for Livorno group. The results showed similar values for Siciliana and Livorno egg quality, highlighting several valuable quality traits from these breeds which might be taken into account for conservation programs.

The aim of the study was to assess the inclusion of different levels of olive cake in pigs' diet as a strategy to replace conventional ingredients and to improve meat quality traits. Seventy-two Pietrain pigs, during the growing-finishing period (50-120 kg BW), were fed with three dietary treatments that contained or did not contain olive cake: 0% (Ctrl), 5% (Low), and 10% (High). The trial lasted 90 days. Weekly, individual body weight (BW) and feed intake (FI) were recorded to calculate average daily gain (ADG) and feed conversion ratio (FCR). At slaughter, the dressing percentage was calculated and carcass weight and backfat thickness were measured. On a section of Longissimus thoracis muscle (LT), pH, color, chemical, and fatty acid composition were determined. Fatty acid profile was also determined in backfat. The statistical model included the effects of diet (Ctrl, Low, and High). The inclusion of 5% of olive cake in the diet improved significantly ( p < 0.05) BW and FCR. Both levels of inclusion (5% and 10%) significantly reduced ( p < 0.05) backfat thickness and intramuscular fat and modified their fatty acid composition, increasing ( p < 0.05) the concentration of MUFA and PUFA and improving ( p < 0.05) quality indices. Results suggest that olive cake did not negatively affect the productive performances.