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Many patients with B-cell malignancies can be successfully treated, although tumor eradication is rarely achieved. T-cell-directed killing of tumor cells using engineered T-cells or bispecific antibodies is a promising approach for the treatment of hematologic malignancies. We investigated the efficacy of CD19xCD3 DART bispecific antibody in a broad panel of human primary B-cell malignancies. The CD19xCD3 DART identified 2 distinct subsets of patients, in which the neoplastic lymphocytes were eliminated with rapid or slow kinetics. Delayed responses were always overcome by a prolonged or repeated DART exposure. Both CD4 and CD8 effector cytotoxic cells were generated, and DART-mediated killing of CD4+ cells into cytotoxic effectors required the presence of CD8+ cells. Serial exposures to DART led to the exponential expansion of CD4 + and CD8 + cells and to the sequential ablation of neoplastic cells in absence of a PD-L1-mediated exhaustion. Lastly, patient-derived neoplastic B-cells (B-Acute Lymphoblast Leukemia and Diffuse Large B Cell Lymphoma) could be proficiently eradicated in a xenograft mouse model by DART-armed cytokine induced killer (CIK) cells. Collectively, patient tailored DART exposures can result in the effective elimination of CD19 positive leukemia and B-cell lymphoma and the association of bispecific antibodies with unmatched CIK cells represents an effective modality for the treatment of CD19 positive leukemia/lymphoma.

In the autumn of 2014, a series of rainfall events affected several sectors of the Liguria region, triggering many shallow landslides and causing three casualties and severe structural damages. The most intensely unstable area covered 385 km2, in which more than 1600 landslides have been identified. After these events, an airborne Light Detection and Ranging survey was carried out. The survey yielded a high-resolution digital terrain model (DTM) and aerial images that provided a means of identifying and mapping all the occurred landslides. The distribution analysis of slope instabilities highlighted the link with various human activities. In fact, the majority of the detected landslides occurred in man-modified areas. Geospatial and statistical analyses provided the identification of three main anthropic factors: terraces, their level of maintenance and road network. Moreover, they quantified their role in landslide triggering. These factors were not analysed as separate elements, but as a continuous process, overlapping in time, in man-made influence on landscape. The identification of such factors is a key element for a correct behaviour characterization of this landscape towards extreme flash floods events.

Exploration of the body representation system (BRS) from kinaesthetic illusions in fMRI has revealed a complex network composed of sensorimotor and frontoparietal components. Here, we evaluated the degree of maturity of this network in children aged 7–11 years, and the extent to which structural factors account for network differences with adults. Brain activation following tendon vibration at 100 Hz (‘illusion’) and 30 Hz (‘no illusion’) were analysed using the two-stage random effects model, with or without white and grey matter covariates. The BRS was already well established in children as revealed by the contrast ‘illusion’ vs ‘no illusion’, although still immature in some aspects. This included a lower level of activation in primary somatosensory and posterior parietal regions, and the exclusive activation of the frontopolar cortex (FPC) in children compared to adults. The former differences were related to structure, while the latter difference reflected a functional strategy where the FPC may serve as the ‘top’ in top-down modulation of the activity of the other BRS regions to facilitate the establishment of body representations. Hence, the development of the BRS not only relies on structural maturation, but also involves the disengagement of an executive region not classically involved in body processing. Keywords: Body representation system, Proprioception, fMRI, DTI, VBM, Children

Background This non-interventional retrospective study, is intended to analyze toxicity and effectiveness of venetoclax of a large cohort of R/R and de novo AML patients treated in Italy since 2015 outside clinical trials with the ultimate goal of improving the knowledge related to venetoclax treatment in the real-life setting. Interim analysis on first 59 patients enrolled is presented here. Aims The aim of this study is to collect data on safety and efficacy of venetoclax in a large cohort of AML patients treated in Italy from 2015 to 2020 in a real-life setting (out of clinical trials). Methods This is a multicenter, retrospective, observational study. All patients with AML treated outside clinical trials with venetoclax as single agent or in combination with other drugs from 1 January 2015 to 1 April 2020 in 40 Italian Hematology Units will be considered for enrollment. Data are collected in accordance with GCP and Helsinky declaration. For this interim analysis data registered into eCRF at 30 June 2020 have been considered. Adverse events (AEs) are graded according to CTCAE v4.03. Survival is estimated with Kaplan-Meyer method. Results The study started on August 2019 and will be completed on August 2021. Up to 30 June 2020 (first data cut off) 30 sites have been activated and 59 AML patients have been registered, 27 males (46%) and 32 females (54%), with a median age of 64 years (range 35 - 83). At the time of venetoclax initiation, 14/59 (24%) patients had a refractory disease, 31 (52.5%) had relapsed disease (14 first relapse and 17 a second or further relapse), eight patients (15%) were newly diagnosed and received venetoclax as first line therapy while in six patients disease status was missing at the time of data cut off. At diagnosis, 52 patients (88%) had bone marrow involvement, of which four (7%) had extramedullary involvement (skin or subcutaneous and deep muscular localization) and one had CNS involvement. 35/59 (59%) patients were classified as fit for intensive chemotherapy, 16 (27%) unfit and two (3%) frail according to SIE, SIES and GITMO group. For six patients fitness status was missing at the time of data cut off. 45/59 (76%) patients had received previous therapy for AML, eight patients were treatment naïve at the time of venetoclax initiation. while for six patients data regarding previous AML therapy were missing at the time of data cut off. In the R/R setting (n=45), the median number of previous therapy lines were 2 (range 1-7). Nine patients had relapse after stem cell transplantation (SCT) and one of them had received double transplant. For 42/45 R/R AML patients the data regarding treatment with venetoclax were available at the time of data cut off. Three out of 42(7%) patients received venetoclax as single agents, 32 (76%) in combination with HMAs (31 Azacitidine and 1 Decitabine), and six with high or intermediate dose ARA-C. In the cohort of R/R patients treated with combination of venetoclax and HMA, in 29/32 patients venetoclax was started as a ramp up phase. The median number of venetoclax cycles was 2 (range 1-13) and the median dose administered was 400 mg daily (range 100-600). Regarding toxicity, 72 adverse events (AEs) were recorded of which 49 were grade III-IV (39 hematologic toxicities, 4 pneumonia, 2 sepsis and 4 other) and 2 fatal (sepsis). For 25 out of 32 patients treated with venetoclax and HMAs combination, a response evaluation was available at the time of data cut off. 15 patients had an evaluable response within 2 months, 7 within 4-months and for 3 the date was missing. 14 (56%) patients obtained composite complete remission (CR=9; 5 =CRi), three patients had a partial response (PR), one stable disease (SD) while seven patients were refractory. Overall, six out of 32 (19%) patients underwent allogenic transplantation after venetoclax-HMAs combination. Median OS of R/R patients treated with venetoclax plus HMA was 182 days (95% C.I 85-421.). Conclusions These preliminary data show that venetoclax in combination with HMAs has an acceptable toxicity profile and is effective in this setting of R/R patients with unfavorable prognosis. Data from more than 100 patients treated in real life setting with venetoclax in Italy since 2015 is expected by the end of 2020, and further analysis on RR patients is ongoing. Data on newly diagnosed patients treated with venetoclax alone or in combination with HMA or chemotherapy will be analyzed. *G.M. and C.C. contributed equally as last author Disclosures Todisco: Jannsen, Abbvie, Jazz:Membership on an entity's Board of Directors or advisory committees.Fracchiolla:Amgen:Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel, accommodations, expenses, Speakers Bureau;ABBVIE:Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel, accommodations, expenses;Gilead:Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel, accommodations, expenses, Speakers Bureau;Pfizer:Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel, accommodations, expenses, Speakers Bureau.Papayannidis:Abbvie, Janssen, Novartis, Amgen, Pfizer:Membership on an entity's Board of Directors or advisory committees, Patents & Royalties.Martelli:Pfizer:Membership on an entity's Board of Directors or advisory committees;Novartis:Membership on an entity's Board of Directors or advisory committees;Celgene:Membership on an entity's Board of Directors or advisory committees;Janssen:Membership on an entity's Board of Directors or advisory committees;AbbVie:Membership on an entity's Board of Directors or advisory committees;Amgen:Membership on an entity's Board of Directors or advisory committees;Jazz Pharmaceuticals:Membership on an entity's Board of Directors or advisory committees.Di Renzo:BerGenBio ASA:Research Funding.Tarella:ADC Therapeutics:Membership on an entity's Board of Directors or advisory committees, Research Funding;ImmunoGen:Research Funding;TG-therapeutics:Research Funding.Rossi:Jazz:Membership on an entity's Board of Directors or advisory committees;Abbvie:Membership on an entity's Board of Directors or advisory committees;Astellas:Membership on an entity's Board of Directors or advisory committees;Novartis:Other: Advisory board;Amgen:Honoraria;Pfizer:Membership on an entity's Board of Directors or advisory committees;Janssen:Membership on an entity's Board of Directors or advisory committees;Daiichi Sankyo:Consultancy, Honoraria;Takeda:Honoraria, Membership on an entity's Board of Directors or advisory committees;Alexion:Membership on an entity's Board of Directors or advisory committees;Celgene:Membership on an entity's Board of Directors or advisory committees;Sanofi:Honoraria.

Background: Acute lymphoblastic leukemia (ALL) is a molecularly heterogeneous disease originating from clonal proliferation of precursor B-lineage cells. In adults, ALL diagnosis is still associated with a dismal prognosis due to the lack of specific targeted therapies. This study was designed to investigate the expression of interleukin-2 receptor alpha chain CD25 in B-ALL and its biological significance, especially following the availability of specific CD25 targeting compounds. Methods:The expression of IL2RA (CD25 gene) was detected by flow cytometry (FC), immunohistochemistry and Western blot analysis, in 25 newly diagnosed ALL patients, both Philadelphia positive (12 patients) and Philadelphia negative (13 patients). Similarly, CD25 expression was assessed in four B-ALL commercially available cell lines. Infection with shRNA specifically directed against CD25 was used to evaluate apoptosis induction and cell cycle arrest in primary B-ALL cells established from two patients. Results:Our data suggest that ALL, and in particular Ph-positive ALL, aberrantly expresses the interleukin-2 receptor alpha chain, CD25. Whereas normal B cells display low amounts of CD25, primary ALL cells and ALL cell lines (over)-express CD25. While the high frequency of CD25 on the surface of many different hematological tumor cells has been established and confirmed in our study, there is little investigation focusing on the significance of CD25 expression. Indeed, CD25 may be present on ALL cells and enable oncogenic signaling pathways. In such respect, we observed that CD25 silencing in primary cells promotes cell cycle arrest and apoptosis induction. While these data support the rational to target CD25, ALL cells did not appear to be in-vitro sensitive to basiliximab, an antibody able to target the Il2RA, but in-vivo investigations are needed to better assess the effects of this therapeutic approach in ALL context. Conclusions:We concluded that CD25 expression is elevated in patients with B-ALL. Our results also demonstrate that CD25 silencing induces cell cycle arrest and apoptosis. The latter result has important implications from a therapeutic point of view. Targeting CD25 receptor with anti-CD25 antibodies or peptide mimetics could be an effective strategy for targeting leukemic cells. Additionally, high CD25 expression could be exploited for the development of CAR-T therapy Disclosures Saglio: Roche:Research Funding;Pfizer:Research Funding;Incyte:Research Funding;Novartis:Research Funding;Ariad:Research Funding;Bristol-Myers Squibb:Research Funding.

Background One-half of all orthopedic surgeries require bone grafting for successful outcomes in fusions, reconstructive procedures, and the treatment of osseous defects resulting from trauma, tumor, infection, or congenital deformity. Autologous bone grafts are taken from the patient's own body and remain the "gold standard" graft choice but are limited in supply and impart significant patient morbidity. Xenograft bone is an attractive alternative from donors with controlled biology, in large supply and at a theoretically lower cost. Clinical results with xenograft bone for orthopedic applications have been mixed in the limited clinical trials published. Methods In the current review, we introduce fundamental principles of bone grafting, systematically review all orthopedic clinical studies reporting outcomes on patients transplanted with xenograft bone, and we present our own clinical results from patients grafted with bovine bone in foot and ankle reconstructive procedures. Results Thirty-one clinical studies were identified for review and the majority (47%) were from spine surgery literature. Favorable results were reported in 44% of studies while 47% of studies reported poor outcomes and discouraged use of xenograft bone products. In our own clinical series, xenograft failed to integrate with host bone in 58% of cases and persistent pain was reported in 83% of cases. Conclusions This is the first systematic review of clinical results reported after bone xenotransplantation for orthopaedic surgery applications. Current literature does not support the use of xenograft bone products and our institution's results are consistent with this conclusion. Our laboratory has reported promising pre-clinical results with a xenograft product derived from porcine cancellous bone, but additional testing is required before considering clinical translation.

Resumen El objetivo del presente trabajo es socializar la mirada docente en torno a las problemáticas detectadas en la población estudiantil del ciclo básico de la carrera Medicina Veterinaria, a partir del análisis de los factores que desde su perspectiva inciden en el recorrido académico de los estudiantes. La instrumentación de una encuesta diseñada para docentes permitió acercarnos a las interpretaciones que realizan de lo que acontece en este tramo de la carrera, con el fin de tomar conciencia de aquellos factores que tienden a repercutir significativamente en el desempeño de los jóvenes. Los resultados obtenidos nos indican que los docentes observan dificultades vinculadas a la interpretación de las consignas, las cuales son atribuidas principalmente a la escasa preparación que han obtenido en el nivel secundario, así como también a factores emocionales que tienden a impactar en la continuidad de los estudios. A su vez, expresan que la desorientación vocacional resulta ser un signo importante que repercute en la motivación por la carrera elegida. Otros indicadores importantes refieren a la falta de métodos de estudios propios, y a dificultades para la expresión oral y escrita. Además, señalan que las propuestas pedagógicas ofrecidas no siempre satisfacen las necesidades de los estudiantes actuales.

Structure from Motion (SfM) is a powerful tool to provide 3D point clouds from a sequence of images taken from different remote sensing technologies. The use of this approach for processing images captured from both Remotely Piloted Aerial Vehicles (RPAS), historical aerial photograms, and smartphones, constitutes a valuable solution for the identification and characterization of active landslides. We applied SfM to process all the acquired and available images for the study of the Champlas du Col landslide, a complex slope instability reactivated in spring 2018 in the Piemonte Region (north-western Italy). This last reactivation of the slide, principally due to snow melting at the end of the winter season, interrupted the main road used to reach Sestriere, one of the most famous ski resorts in north-western Italy. We tested how SfM can be applied to process high-resolution multisource datasets by processing: (i) historical aerial photograms collected from five diverse regional flights, (ii) RGB and multi-spectral images acquired by two RPAS, taken in different moments, and (iii) terrestrial sequences of the most representative kinematic elements due to the evolution of the landslide. In addition, we obtained an overall framework of the historical development of the area of interest, and distinguished several generations of landslides. Moreover, an in-depth geomorphological characterization of the Champlas du Col landslide reactivation was done, by testing a cost-effective and rapid methodology based on SfM principles, which is easily repeatable to characterize and investigate active landslides.

The potential of satellite-based interferometry for landslides activity definition is well known. Many applications have demonstrated that this technique represents a strong supporting tool for landslide risk assessment and monitoring, at both local and regional scale. The ESA' Sentinel-1 (S1) constellation acquires with high revisit-time in a conflict-free operational mode, timely distributing data without costs or restrictions. All these characteristics allow relying upon S1 service for a long-term operational and sustainable use, as a support of both disaster risk reduction and response. For this reason, more and more research projects are based on the use of S1 in geohazards monitoring and early warning. The aim of this work is to present the results achieved in the framework of the European Project (ECHO) U-Geohaz regarding landslides monitoring and early warning supporting tools based on S1. The study area of the project is the Valle d'Aosta region (VDA) that is widely affected by landslide process of different size and typology. Specifically, the Deep-Seated Gravitational Slope Deformations (DSGSDs) are the most representative in terms of area coverage. The study of the state of activity and evolution of these phenomena is very important because DSGSDs affect entire valley flanks and several villages have been built on them or nearby. The use of S1-based interferometry can potentially give a near-real time information on their activity over the whole region supporting decision makers and early warning systems. At the same time, VDA presents challenging characteristics for what concerns the radar response and thus the obtainable results. The main limiting factor is the snow coverage, which is also strongly related to the topography. The topographic relief is highly variable, ranging from 300 m a.s.l. to peaks higher than 4000 m a.s.l. with steep slopes. The regional climate is characterized by wide range of temperatures varying a lot from the mountainous zone to the bottom of the valleys, implying also a strong variation in snow precipitations. This strong variation, in space and time, affects the DInSAR results in terms of coherence and spatial coverage. In this context, a strong effort has been done to develop a processing approach to improve the spatial and temporal sampling, without losing the quality of the results. Moreover, a method that extracts the most significant Active Deformation Areas (ADA) in a semi-automatic way has been adapted and applied. The main results will be exposed.

Adoptive transfer of T lymphocytes (ACT) engineered with T-cell receptors (TCRs) of known antitumor specificity is an effective therapeutic strategy. However, a major constraint of ACT is the unpredictable interference of the endogenous TCR α and β chains in pairing of the transduced TCR. This effect reduces the efficacy of the genetically modified primary T cells and carries the risk of generating novel TCR reactivities with unintended functional consequences. Here, we show a powerful approach to overcome these limitations. We engineered TCR α and β chains with mutations encompassing a conserved motif (FXXXFXXS) required to stabilize the pairing of immunoglobulin heavy chain transmembrane domains. Molecular modeling supported the preferential pairing of mutated TCR and impaired pairing between mutated and wild-type TCRs. Expression of the mutated TCR was similar to wild type and conferred the expected specificity. Fluorescence resonance energy transfer analysis in mouse splenocytes transduced with mutated or wild-type TCRs showed a higher proximity of the former over the latter. Importantly, we show that mutated TCRs effectively outcompete endogenous TCRs and improve in vitro antitumor cytotoxicity when expressed in ex vivo isolated human T cells. This approach should contribute to improving current protocols of anticancer immunetherapy protocols.

In acute myeloid leukemia (AML), blasts lose their ability to differentiate into mature cells and to undergo apoptosis. Accordingly, a proapoptotic and differentiating therapy (arsenic and retinoic acid) has dramatically improved survival in acute promyelocytic leukemia; however, a similar combination therapy is not available for other AML subtypes. In 2016, inhibition of dihydroorotate dehydrogenase (DHODH), a key enzyme of the pyrimidine biosynthesis, was found to induce differentiation in several AML models; for in vivo studies, brequinar (BRQ) was utilized. Starting from BRQ and applying a scaffold-hopping replacement, we have recently developed a new DHODH inhibitor, Meds433, which could induce differentiation at a 1-log lower concentration compared to BRQ (Sainas, J Med Chem 2018). Here we characterize Meds433 with in vitro and in vivo experiments, showing that it has a significant pro-apoptotic effect in several AML cell lines, which is at least partially independent from the differentiating effect. Analyzing the kinetic of differentiation induced by Meds433 on U937 and THP1 cell lines, and comparing the data with the number of viable cells, we noticed that cells started to die before the differentiation effect could be significant. Hence, we decided to investigate the proapoptotic effect of Meds433, treating several AML (U937, THP1, OCI-AML3, NB4) and non-AML (CEM, P3j, peripheral blood mononuclear cells-PBMC) cell lines with Meds433, and analyzing the expression of Annexin V and propidium in flow cytometry. Experiments demonstrated that Meds433 had a significant pro-apoptotic effect on several AML cell lines (Fig.1), but not on non-AML cell lines. The apoptotic rate increased with the time of exposure (3 vs 6 days), allowing to obtain a good apoptotic rate also in OCI-AML3, the most resistant cell line in our hands. As for the differentiation experiments, Meds433 could induce apoptosis at a 1-log inferior concentration compared to BRQ. More interestingly, in NB4 cells, a strong apoptotic effect was not associated with any differentiating feature, indicating that DHODH inhibition can induce apoptosis directly. As NB4 is a promyelocytic cell line, we also compared the effects of ATRA (all-trans retinoic acid), Meds433 and their combination. ATRA alone was found to induce strong differentiation and mild apoptosis on NB4 cells, while Meds433 alone could induce exclusively apoptosis; finally, the combination of ATRA and Meds433 increased both the rate of differentiating and apoptotic cells compared to ATRA only. We next tried to further characterize the apoptotic effect. When experiments were performed in the presence of uridine, a downstream product of DHODH in the pyrimidine biosynthesis, the apoptotic effect was totally abrogated. This phenomenon was already observed in the differentiation experiments (Sainas, J Med Chem 2018), and it confirms that both differentiation and apoptosis are indeed caused by the pyrimidine depletion rather than off-target mechanisms. Moreover, when experiments were performed in hypoxic conditions, the rate of apoptosis did not change, suggesting that Meds433 could work in the bone marrow hypoxic niche of (leukemic) stem cells. Further analyzing the effect of Meds433 on non-AML cells, we evaluated the maturation of T-lymphocytes, from naïve to TEMRA (T effector memory RA), finding no influence at all. Finally, preliminary results from in vivo experiments show that i) Meds433 is not toxic on Balb/c mice after 5 weeks of intraperitoneal administration; ii) the half-life is limited to 4-6 hours and iii) Meds433 has a good antileukemic activity (approximately 50% reduction of the tumor volume compared with control, after an 18-day treatment of THP1-xenograft in NSG mice). In conclusion, our work demonstrates that: i) DHODH inhibition can induce apoptosis in AML cells both directly and as a result of differentiation, partially depending on the cell line; ii) Meds433 is a novel, promising antileukemic drug, with limited toxicity, which could be active on a wide variety of AML. Since continuous exposure to the drug is fundamental in the pyrimidine starvation strategy, we are currently optimizing Meds433 pharmacokinetic profile in order to maximize the in vivo antileukemic activity. Disclosures Saglio: BMS: Consultancy; Novartis: Consultancy; Ariad: Consultancy; Incyte: Consultancy; Pfizer: Consultancy; Jansen: Consultancy; Celgene: Consultancy.

Cytokine-induced killer (CIK) cells consist of a heterogeneous population of polyclonal T lymphocytes displaying NK phenotype and HLA-unrestricted cytotoxic activity against a broad range of tumors. We sought to determine whether transduction of CIK cells with T cell receptor (TCR) genes specific for tumor-associated antigens could generate effector cells endowed with a double mechanism of tumor recognition. HLA-A2-restricted TCR-transduced (TD) CIK directed against the melanoma antigens Mart1 and NY-ESO1 were generated by lentiviral transduction and successfully expanded over a 3-4-week period. TD-CIK cells were both CD3(+)/CD56(-) and CD3(+)/CD56(+) (31±8% and 59±9%, respectively), indicating that both major histocompatibility complex (MHC)-restricted T cells and MHC-unrestricted CIK could be targeted by lentiviral transduction. At the end of the culture, the majority of both unmodified and TD-CIK displayed an effector memory phenotype, without considerable expression of replicative senescence and exhaustion markers. Functionally, TD-CIK specifically recognized tumor cells expressing the relevant antigen as well as maintained their MHC-unrestricted tumor activity. The cytotoxic activity of TD-CIK against HLA-A2(+) melanoma cell lines was significantly higher than the untransduced counterparts at a low effector:target ratio (cytotoxic activity of TD-CIK was from 1.9- to 4.3-fold higher than untransduced counterparts). TD-CIK were highly proficient in releasing high amount of IFN-γ upon antigen-specific stimulation and were able to recognize primary melanoma targets. In conclusion, we showed that (1) the reproducibility and simplicity of CIK transduction and expansion might solve the problem of obtaining adequate numbers of potent antitumor effector cells for adoptive immunotherapy; (2) the presence of both terminal effectors as well as of less differentiated progenitors might confer them long survival in vivo; and (3) the addition of an MHC-restricted antigen recognition allows not only targeting tumor surface antigens but also a wider range of cytoplasmic or nuclear antigens, involved in tumor proliferation and survival. TD-CIK cells with a double mechanism of tumor recognition are an attractive and alternative tool for the development of efficient cell therapeutic strategies.

Combined direct antineoplastic activity and the long-lasting immunological effects of allogeneic hematopoietic cell transplant (HCT) can cure many hematological malignancies, but broad adoption requires non-relapse mortality (NRM) rates and graft-versus-host disease (GVHD) control. Recently, posttransplant cyclophosphamide (PTCy) given after a bone marrow transplant significantly reduced GVHD-incidence, while PTCy given with tacrolimus/mofetil mycophenolate (T/MMF) showed activity following allogeneic peripheral blood stem cell transplantation (alloPBSCT). Here, we report the experience of a larger cohort (85 consecutive patients) and expanded follow-up period (03/2011–12/2019) with high-risk hematological malignancies who received alloPBSCT from Human-Leukocyte-Antigens HLA-matched unrelated/related donors. GVHD-prophylaxis was PTCy 50 mg/kg (days+3 and +4) combined with T/MMF (day+5 forward). All patients stopped MMF on day+28 with day+110 = median tacrolimus discontinuation. Cumulative incidences were 12% for acute and 7% for chronic GVHD- and no GVHD-attributed deaths. For surviving patients, the 12, 24, and 36-month probabilities of being off immunosuppression were 92, 96, and 96%, respectively. After a 36-month median follow-up, NRM was 4%, median event-free survival (EFS) and overall survival (OS) had yet to occur. One- and two-year chronic GVHD-EFS results were 57% (95% CI, 46–68%) and 53% (95% CI, 45–61%), respectively, with limited late infections and long-term organ toxicities. Disease relapse caused the most treatment failures (38% at 2 years), but low transplant toxicity allowed many patients (14/37, 38%) to receive donor lymphocyte infusions as a post-relapse strategy. We confirmed that PTCy+T/MMF treatment effectively prevented acute and chronic GVHD and limited NRM to unprecedented low rates without loss of disease control efficacy in an expanded patient cohort. This trial is registered at U.S. National Library of Medicine as #NCT02300571.

How neural specificity for distinct conceptual knowledge categories arises is central for understanding the organization of semantic memory in the human brain. Although there is a large body of research on the neural processing of distinct object categories, the organization of action categories remains largely unknown. In particular, it is unknown whether different action categories follow a specific topographical organization on the cortical surface analogously to the category-specific organization of object knowledge. Here, we tested whether the neural representation of action knowledge is organized in terms of nonsocial versus social and object-unrelated versus object-related actions (sociality and transitivity, respectively, hereafter). We hypothesized a major distinction of sociality and transitivity along dorsal and ventral lateral occipitotemporal cortex (LOTC), respectively. Using fMRI-based multivoxel pattern analysis, we identified neural representations of action information associated with sociality and transitivity in bilateral LOTC. Representational similarity analysis revealed a dissociation between dorsal and ventral LOTC. We found that action representations in dorsal LOTC are segregated along features of sociality, whereas action representations in ventral LOTC are segregated along features of transitivity. In addition, representations of sociality and transitivity features were found more anteriorly in LOTC than representations of specific subtypes of actions, suggesting a posterior-anterior gradient from concrete to abstract action features. These findings elucidate how the neural representations of perceptually and conceptually diverse actions are organized in distinct subsystems in the LOTC.The lateral occipitotemporal cortex (LOTC) is critically involved in the recognition of objects and actions, but our knowledge about the underlying organizing principles is limited. Here, we discovered a dorsal-ventral distinction of actions in LOTC: dorsal LOTC represents actions based on sociality (how much an action is directed to another person) in proximity to person knowledge. In contrast, ventral LOTC represents actions based on transitivity (how much an action involves the interaction with inanimate objects) in proximity to tools/artifacts in ventral LOTC, suggesting a mutually dependent organization of actions and objects. In addition, we found a posterior-to-anterior organization of the LOTC for concrete and abstract representations, respectively. Our findings provide important insights about the organization of actions in LOTC.

The introduction of different generations of tyrosine kinase inhibitors (TKIs) significantly improved outcome and survival rate in chronic myeloid leukemia (CML) patients. However, long-term use of TKIs is concomitant with many side effects that affect the quality of life in patients. Approximately half of CML patients achieve deep molecular response (DMR), this makes them suitable candidates to discontinue the TKI therapy in a controlled condition, and about half of them will remain in treatment free remission (TFR) after discontinuation. It has been shown that a small population of leukemia stem cells (LSCs) as the residual disease burden is present at diagnosis, during the treatment, and in patients who are in TFR. While CML LSCs have many features in common with HSCs, they express specific markers such as CD25, CD26, IL1-RAP, etc., which can be used for the diagnosis and targeting. Protection by the bone marrow microenvironment and activity of signaling pathways such as WNT/β catenin, Hedgehog, PI3K, JAK/STAT in CML LSCs in a BCR-ABL dependent and independent manner guarantee their survival and elimination of these cells solely using TKIs seems ineffective. Herein we designed a pegylated liposomal nanocarrier conjugated with a specific antibody against CD26 (Begelomab, ADIENNE, Lugano, Switzerland). Then we loaded this immunoliposome with venetoclax, a BCL2 inhibitor, to eliminate CML LSCs selectively and to spare normal HSCs. First, we measured the expression of CD26 in the bone marrow and peripheral blood samples of newly diagnosed patients. We had a high expression of CD26 in CD34+/CD38- of both PB and BM, and a low expression on CD34+/CD38+ (progenitors) cells. Also, the expression of this marker in resistant patients to TKIs was visible while it was absent in normal stem cells. After the synthesis of the liposome, we conjugated Begelomab to the liposome. Then, we tested the selectivity of the designed system in different positive and negative cells. Our designed immunoliposome showed a strong selectivity toward CD26 positive cells. We also tested the selectivity on CML primary cells; in particular, we sorted newly diagnosed CML samples based on CD34+/CD38-/CD26- for HSCs and CD34+/CD38-/CD26+ for LSCs. Based on the confocal and flow cytometry analysis, our designed immunoliposome selectively targets LSCs and spares HSCs. Then we loaded this immunoliposome with venetoclax, and we treated CD26 positive and negative cells with this system. Based on our preliminary results, this immunoliposome loaded venetoclax specifically induced apoptosis in CD26+ cells, with higher activity compared to free venetoclax at the same dose. However, more analysis will be performed to confirm the selectivity of this system. Based on the obtained results, CD26 in newly diagnosed CML patients is expressed by CML LSCs and is a suitable option for diagnosis and targeting. Our preliminary data strongly suggest that we can selectively target CML LSCs. The main advantage of this system is its precision to hit the target. So we expect that after the drug release, the LSCs will be eliminated without any side effects on normal cells. Liposomes are suitable carriers because of their biocompatibility, self-assembly, large drug payload, and minimal toxicity. This strategy may help us to increase the number of patients attaining and maintaining TFR without relapsing. Disclosures Saglio: Ariad: Research Funding; Pfizer: Research Funding; Incyte: Research Funding; Roche: Research Funding; Bristol-Myers Squibb: Research Funding; Novartis: Research Funding.

Deep-seated Gravitational Slope Deformations (DsGSDs) are huge ground-deformation slow evolving phenomena, highly widespread in alpine territory. Their long-lasting evolution, with continuous deformation rate, may represents a natural hazard, able to endanger various anthropic structures and infrastructures. Until today, the development of technical and regulatory tools, aimed to effectively manage the interactions between DsGSDs and anthropic elements, has been generally lightly considered in risk management and land use planning. The definition of the type and severity of impacts on the anthropic elements, becomes increasingly important in terms of urban planning and risk management, and deserve an update in the current adopted procedures. Focusing on the Western Italian Alps, we implemented an interdisciplinary analysis, based on multi-source data, by means of geoinformatics, remote sensing and archive consultation approaches. Intersecting DsGSDs available information with the urbanized territory in a Geographic Information System environment, we obtained, despite the high data heterogeneity, an overall framework of the existing interactions. Specifically, we defined the interactions between these large phenomena and buildings, roads and rail networks, and linear infrastructures, as penstocks, waterworks or dams, also highlighing the state of activity of the inventoried phenomena. Moreover, we analysed the degree of the DsGSD impacts on the anthropic elements, detecting and classifying all the documented damages within the Italian Western Alps territory. The obtained results highlight the need for an innovative approach in DsGSDs risk assessment, both in terms of the definition of their behavior over time and of their impacts on the anthropic elements, for a more effective land use planning and a proper handling of these phenomena in the legislation framework.

Context The inhibition of Dihydroorotate Dehydrogenase (DHODH) was recently found to induce differentiation and apoptosis in acute myeloid leukemia (AML). Multiple clinical trials testing DHODH inhibitors are currently ongoing, but the clinical effectiveness of these compounds is expected to be lower than in vitro due to: i) the heterogeneity of the AML genetic landscape and the well-known ability of leukemic cells to undergo clonal escape when treated with single agents, and ii) the pyrimidine salvage pathway, which could mitigate the pyrimidine starvation induced by DHODH inhibitors. Objective To increase the apoptotic rate induced by DHODH inhibitors in AML cells by testing multiple associations both with classical chemotherapeutical agents and targeted compounds, leveraging the synthetic lethality concept. Design MEDS433, the DHODH inhibitor developed by our group, was tested on several AML cell lines (THP1, MV4-11, NB4, U937, OCI-AML3) and primary cells, alone and in combination with cytarabine, anthracyclines, and decitabine. Moreover, in order to block the pyrimidine salvage pathway, MEDS433 was also combined with dipyridamole, a potent inhibitor of hENT1 and hENT2 nucleotide/nucleoside transport channels. Results The association of MEDS433 with cytarabine, idarubicin, and decitabine resulted in an additive effect, with a significant increase of the apoptotic rate in all tested AML cell lines. Moreover, the combination of MEDS433 with dipyridamole resulted in a synergistic effect, with an apoptotic rate ranging between 69% and 95% after just 3 days of treatment at the lowest effective MEDS433 concentration (0.1 μM), depending on the cell line. The combination of MEDS433, dipyridamole, and chemotherapy resulted in the massive death of all AML cells (>90%). The best associations were evaluated on primary cells, confirming optimal results. Conclusions The addition of DHODH inhibitors to the (3+7) regimen or to hypomethylating agents could significantly increase the performances of these classical treatments. Moreover, a deep pyrimidine starvation obtained through the inhibition of both DHODH and the pyrimidine salvage pathway, could further increase the antileukemic activity. Ongoing in vivo experiments will be determinant to verify this hypothesis and to assess the toxicity on normal cells. This work was supported by AIRC (IG-23344).

The immunophenotype is a key element to classify B-cell Non-Hodgkin Lymphomas (B-NHL), while it is routinely obtained through immunohistochemistry, the use of flow cytometry (FC) could bear several advantages. However, few FC laboratories can rely on a long-standing practical experience, and the literature in support is still limited, as a result, the use of FC is generally restricted to the analysis of lymphomas with bone marrow or peripheral blood involvement. In this work, we applied machine learning to our database of 1465 B-NHL samples from different sources, building four artificial predictive systems which could classify B-NHL in up to nine of the most common clinico-pathological entities. Our best model shows an overall accuracy of 92.68%, a mean sensitivity of 88.54% and a mean specificity of 98.77%. Beyond the clinical applicability, our models demonstrate (i) the strong discriminatory power of MIB1 and Bcl2, whose integration in the predictive model significantly increased the performance of the algorithm, (ii) the potential usefulness of some non-canonical markers in categorizing B-NHL, and (iii) that FC markers should not be described as strictly positive or negative according to fixed thresholds, but they rather correlate with different B-NHL depending on their level of expression.

Active landslide risk assessment and management are primarily based on the availability of dedicated studies and monitoring activities. The establishment of decision support for the efficient management of active landslides threatening urban areas is a worthwhile contribution. Nowadays, consistent information about major landslide hazards is obtained through an interdisciplinary approach, consisting of field survey data and long-time monitoring, with the creation of a high populated dataset. Nevertheless, the large number and variety of acquired data can generate some criticalities in their management. Data fragmentation and a missing standard format of the data should represent a serious hitch in landslide hazard management. A good organization in a standard format can be a good operative solution. Based on standardized approaches such as the ICAO (International Civil Aviation Organization), we developed a standard document called operative monography. This document summarizes all available information by organizing monitoring data and identifying possible lacks. We tested this approach in the Aosta Valley Region (NW Italy) on five different slow moving landslides monitored for twenty years. The critical analysis of the available dataset modifies a simple sequence of information in a more complex document, adoptable by local and national authorities for a more effective management of active landslides.

Purpose Obesity and excessive weight gain during pregnancy is a growing problem, conferring severe health risks for both mother and fetus. Exercise can help combat this epidemic. However, many pregnant women are not meeting the American Congress of Obstetricians and Gynecologists’ (ACOG’s) 2015 guidelines for exercise during pregnancy. The objective of this study was to evaluate obstetricians’ beliefs and recommendations regarding exercise during pregnancy compared to ACOG’s 2015 recommendations. Method Obstetricians were recruited via three different forums to complete a twenty-question survey: at a regional conference for Alabama and Mississippi ACOG members, at the University of Alabama at Birmingham’s Obstetrics and Gynecology Department’s Grand Rounds, and via telephone. Univariate statistical analysis was conducted with RedCap. Results Seventy-one surveys were completed: 33 from the ACOG conference, 27 from Grand Rounds, and 11 from those recruited by telephone. Eighty-eight percent (n=60) of respondents correctly identified ACOG’s recommendation of unrestricted exercise for women with uncomplicated pregnancies. One-fourth (24%; n=16) regularly discuss exercise with most (76%-100%) pregnant patients. Most (57%; n=59) do not consistently (“never,” “rarely,” or “sometimes”) recommend sedentary patients begin exercising during pregnancy. Nearly all (97%; n=66) advise first-trimester patients to perform aerobic exercise two to five days per week, but the recommended duration varies. One-fourth (24%; n=16) do not recommend strength-training exercise during the first trimester. Twenty-five percent (n=17) and 32% (n=22) recommend decreased aerobic or strength-training exercise, respectively, in the third trimester. More than half (54%; n=37) recommend pregnant patients limit exercise by heart rate, most commonly 121-140 bpm (25%; n=17) or 141-160 bpm (24%; n=16). Sixty-eight percent (n=46) feel “comfortable” or “very comfortable” providing advice on exercise during pregnancy. Conclusion Despite believing exercise benefits pregnant women, knowing ACOG’s 2015 guidelines endorse unrestricted exercise for women with uncomplicated pregnancies, and feeling comfortable discussing this topic with patients, obstetricians are not consistently counseling their pregnant patients on exercise. Notably, physicians are not instructing their sedentary pregnant patients to exercise. While most physicians provide appropriate advice on aerobic exercise, their advice on resistance training, maximum heart rate during exercise and third-trimester exercise are often discordant with ACOG’s guidelines.

Allogeneic hematopoietic cell transplantation (allo-HCT) is an adoptive immunotherapy strategy whose effectiveness relies on graft-versus-tumor (GVT) effect. We explored the feasibility of enhancing GVT after allo-HCT by peptide vaccination. Two myeloma patients were transplanted with a fludarabine-total body irradiation conditioning regimen and vaccinated with an HLA-A*0201-restricted modified survivin nonapeptide, plus montanide as adjuvant. At time of first vaccination, one patient had just attained serological remission despite documented relapse after transplant, while the other patient was in stable disease. Both patients had an immune response to vaccination: the frequency of survivin-specific CD8+ T cells increased between second and sixth vaccination and accounted for 0.5–0.8% of CD8+ cells; CD8+ cells were functional in ELISPOT assay. The first patient persists in complete remission with a follow-up of >5 years, while the second patient did not have a clinical response and vaccination was halted. We analyzed the T-cell receptor (TCR) repertoire of the first patient by spectratyping and found that vaccination did not affect the diversity of TCR profile, indicating that survivin clonotypes were probably spread in multiple TCR families. We generated a limited number (n = 4) of survivin-specific T cell clones: three were reactive only against the modified peptide, whereas one clone recognized also the naive peptide. Peptide vaccination is safe and applicable after allo-HCT and elicits an efficient antigen-specific T cell response without causing graft-versus-host disease.

Human dihydroorotate dehydrogenase ( hDHODH) catalyzes the rate-limiting step in de novo pyrimidine biosynthesis, the conversion of dihydroorotate to orotate. hDHODH has recently been found to be associated with acute myelogenous leukemia, a disease for which the standard of intensive care has not changed over decades. This work presents a novel class of hDHODH inhibitors, which are based on an unusual carboxylic group bioisostere 2-hydroxypyrazolo[1,5- a]pyridine, that has been designed starting from brequinar, one of the most potent hDHODH inhibitors. A combination of structure-based and ligand-based strategies produced compound 4, which shows brequinar-like hDHODH potency in vitro and is superior in terms of cytotoxicity and immunosuppression. Compound 4 also restores myeloid differentiation in leukemia cell lines at concentrations that are one log digit lower than those achieved in experiments with brequinar. This Article reports the design, synthesis, SAR, X-ray crystallography, biological assays, and physicochemical characterization of the new class of hDHODH inhibitors.

El presente trabajo busca analizar diversos indicadores vinculados a la práctica evaluativa docente en las diferentes asignaturas que integran el primer semestre del programa de la carrera Medicina Veterinaria de la Facultad de Ciencias Veterinarias de la Universidad Nacional del Litoral, y visibilizar las percepciones que los estudiantes tienen en relación con dicha práctica. Se indagó acerca de las pautas que utilizan los docentes al momento de evaluar y del material de estudio utilizado por los estudiantes para la preparación de exámenes. Los resultados obtenidos permiten reafirmar que la evaluación es un proceso complejo. Este estudio reveló que los estudiantes perciben una falta de consenso en los criterios evaluativos docentes, así como una escasa retroalimentación del proceso realizado. En general, el proceso de aprendizaje estudiantil se complementa con la utilización de estrategias metacognitivas. Con respecto al material de estudio, la mayoría de los estudiantes utilizan apuntes de elaboración propia o confeccionados por las cátedras. El aporte de esta investigación manifiesta la necesidad de construir espacios de debate en torno a las concepciones que los docentes poseen a la hora de evaluar, generando otras miradas que podrían enriquecer los procesos de enseñanza, de aprendizaje y en particular, de las prácticas evaluativas.

Non-infectious soft tissue lesions of the foot and ankle are relatively rare clinically. These include benign and malignant neoplasms, as well as non-neoplastic or pseudotumoral lesions such as ganglionic, synovial and epidermoid cysts, intermetatarsal and adventitious bursitis, inflammatory lesions like gouty tophi and rheumatoid nodules, Morton's neuroma, and granuloma annulare. A 48-year-old male with a history of medically treated tophaceous gout presented with left foot neuropathic pain and paresthesia, in the setting of a well-circumscribed soft tissue lesion of the second intermetatarsal space, suspected to be a Morton's neuroma. Magnetic resonance imaging (MRI) showed a 4.1 x 2.7 x 2.6 cm heterogeneous soft tissue mass containing multiple cystic areas. Excisional biopsy was performed and histologic examination revealed well-circumscribed nodules of amorphous material containing needle-shaped clefts, rimmed by histiocytes, and multinucleated giant cells consistent with a gouty tophus. This is the first case reported in the literature of an intermetatarsal gouty tophus causing neuropathic pain and paresthesia. While Morton's neuroma is the most common cause of this presentation, this case illustrates that other pseudotumoral lesions, such as a gouty tophus, may present similarly, and should be considered in the differential diagnosis. While most cases of tophaceous gout can be adequately treated with urate-lowering therapy, surgery may be indicated for tophi that do not resolve with medical treatment based upon symptom severity, compression of nearby structures, and functional impairment.

La collana pubblica gli atti del convegno annuale di Linguistica Computazionale (CLiC-it), che ha lo scopo di costituire un luogo di discussione di riferimento nel campo delle ricerce sulla linguistica computazionale. Gli atti includono interventi sul trattamento automatico della lingua, comprendenti le riflessioni teoriche e metodologiche sul tema, e forniscono un contributo importante per questo campo di ricerca. Le altre tematiche principali sono: la linguistica computazionale, la linguistica, le scienze cognitive, l'apprendimento automatico, l'informatica, la rappresentazione della conoscenza, l'information retrieval e l'umanistica digitale. L'organizzazione del convegno è il risultato dello sforzo dell'Associazione Italiana di Linguistica Computazionale (AILC http://www.ai-lc.it/), rappresentata ogni anno da alcuni dei membri organizzatori, che sono affiliati anche ad altre organizzazioni che operano nell'ambito della linguistica computazionale. This book presents the proceedings of the annual Conference of Computational Linguistics (CLiC-it), which aims to provide a forum for discussion in the field of computational linguistics. The proceedings include interventions on the automatic treatment of language, with theoretical and methodological reflections on the subject, and provide an important contribution to this field of research. The other main topics are: computational linguistics, linguistics, cognitive sciences, automatic learning, computer science, knowledge representation, information retrieval and digital humanities. The organization of the conference is the result of the effort of the Italian Association of Computational Linguistics (AILC http://www.ai-lc.it/), represented every year by some of the organizing members, who are also affiliated to other organizations operating in the field of computational linguistics.

State-of-the-art Part-of-Speech taggers have been thoroughly evaluated on standard Italian. To understand how Part-of-Speech taggers that have been pretrained on standard Italian fare with a wide array of language anomalies, we evaluate five Part-of-Speech taggers on a corpus of student essays written throughout the largest Italian-speaking area outside of Italy. Our preliminary results show that there is a significant gap between their performance on non-standard Italian and on standard Italian, and that the performance loss mainly comes from relatively subtle tagging errors within morphological categories as opposed to coarse errors across categories. Gli strumenti di Part-of-Speech tagging più rappresentativi dello stato dell’arte sono stati analizzati a fondo con l’italiano standard. Per capire come strumenti pre-addestrati sull’italiano standard si comportano in presenza di un’ampia gamma di anomalie linguistiche, analizziamo le prestazioni di cinque strumenti su di un corpus di elaborati redatti da studenti della scuola dell’obbligo nella Svizzera Italiana. I nostri risultati preliminari mostrano che esiste un notevole divario tra le prestazioni sull’italiano nonstandard e quelle sull’italiano standard, e che la perdita di prestazioni deriva principalmente da errori di tagging relativamente sottili all’interno delle categorie grammaticali.