Effectiveness of enzymatic hydrolysis for reducing the allergenic potential of legume by-products

The interest in agri-food residues and their valorization has grown considerably, and many of them are today considered to be valuable, under-exploited sources of different compounds and notably proteins. Despite the beneficial properties of legumes by-products, there are also some emerging risks to consider, including their potential allergenicity. In this work the immunoreactivity of chickpea, pea, and white bean by-products was assessed, and whether the production of enzymatic hydrolysates can be an effective strategy to reduce this allergenic potential. The results presented clearly indicate that the efficiency of this strategy is strongly related to the enzyme used and the food matrix. All legume by-products showed immunoreactivity towards serum of legume-allergic patients. Hydrolysates from alcalase did not show residual immunoreactivity for chickpea and green pea, whereas hydrolysates from papain still presented some immunoreactivity. However, for white beans, the presence of antinutritional factors prevented a complete hydrolysis, yielding a residual immunoreactivity even after enzymatic hydrolysis with alcalase

Genetic and phenotypic attributes of splenic marginal zone lymphoma

Splenic marginal zone B-cell lymphoma (SMZL) is a heterogeneous clinico-biological entity. The clinical course is variable, multiple genes are mutated with no unifying mechanism, and essential regulatory pathways and surrounding microenvironments are diverse. We sought to clarify the heterogeneity of SMZL by resolving different subgroups and their underlying genomic abnormalities, pathway signatures, and microenvironment compositions to uncover biomarkers and therapeutic vulnerabilities. We studied 303 SMZL spleen samples collected through the IELSG46 multicenter international study (NCT02945319) by using a multiplatform approach. We carried out genetic and phenotypic analyses, defined self-organized signatures, validated the findings in independent primary tumor metadata and in genetically modified mouse models, and determined correlations with outcome data. We identified 2 prominent genetic clusters in SMZL, termed NNK (58% of cases, harboring NF-κB, NOTCH, and KLF2 modules) and DMT (32% of cases, with DNA-damage response, MAPK, and TLR modules). Genetic aberrations in multiple genes as well as cytogenetic and immunogenetic features distinguished NNK- from DMT-SMZLs. These genetic clusters not only have distinct underpinning biology, as judged by differences in gene-expression signatures, but also different outcomes, with inferior survival in NNK-SMZLs. Digital cytometry and in situ profiling segregated 2 basic types of SMZL immune microenvironments termed immune-suppressive SMZL (50% of cases, associated with inflammatory cells and immune checkpoint activation) and immune-silent SMZL (50% of cases, associated with an immune-excluded phenotype) with distinct mutational and clinical connotations. In summary, we propose a nosology of SMZL that can implement its classification and also aid in the development of rationally targeted treatments

IN VITRO CHARACTERISTICS OF HEMATOPOIETIC PROGENITORS FROM PRIMARY MYELOFIBROSIS PATIENTS CORRELATE WITH IPSS/DIPSS RISK CATEGORY

Hyperplasia of morphologically abnormal megakaryocytes (MKs) is a hallmark of primary myelofibrosis (PMF) but the molecular events leading to MK abnormalities are still unclear. We previously demonstrated that, in thrombopoietin (TPO)-stimulated cultures, PMF CD34+ cells showed enhanced in vitro expansion capacity and impaired megakaryocytic differentiation compared to CD34+ cells from healthy individuals, and that the over-expression of the proto-oncogene protein kinase C epsilon (PKC) contributes to these abnormalities (Masselli et al. ASH 2013 abstr.114). Here we investigated whether clinical (included in the IPSS or DIPSS risk category) or biological (JAK2 mutational status) variables might impact the in vitro behavior of TPO-stimulated PMF CD34+ cells. We stratified 8 PMF patients according to the IPSS or DIPSS category (low/intermediate vs high risk) and JAK2 mutational status and evaluated: (1) Fold increase (FI) at day 14 of culture, (2) MK differentiation (% of CD41+ and CD42b+ cells and% of proplatelet-forming MKs) and (3) PKC protein levels (by western blot). CD34+ cells from high risk PMFs displayed increased proliferative capacity as compared to low/intermediate risk (FI: 44±0.2 vs 26.5±4.3, p=0.012), while no difference could be observed between JAK2V617F+ and JAK2V617F- PMFs (FI: 37.2±11.8 vs 27.9±5, p=0.39). Additionally, high risk PMFs revealed impaired MK differentiation potential, as indicated by the lower% of CD41+ and CD42b+ cells (respectively: 26.3±9.4 vs 54.2.6, p=0.008 and 16.1±8 vs 38.2±3, p=0.011) and proplatelet-forming MKs (0.67±0.21 vs 1.8±0.25, p=0.035). By contrast, no statistical difference was observed according to the JAK2 mutation. Finally, we found that high risk patients- derived MKs are characterized by higher expression of PKC as compared to low/intermediate risk ones (relative PKC /GAPDH OD values: 1.75±0.52 in high risk vs 0.82±0.29 in low risk, p=0.023). Conversely, PKC levels were comparable among JAK2V617F+ and JAK2V617F- PMFs (1.11±0.34 vs 1.3±0.8, p=0.72). These data indicate that the degree of in vitro growth and megakaryocytic commitment of PMF CD34+ cells is correlated to the aggressiveness of the disease (indicated by IPSS/DIPSS risk category) and not to the JAK2V617F mutation. Similarly, we found that PKC levels are significantly greater in high vs low/intermediate risk patients, leading us to speculate that PKC can be utilized as a marker of high disease burden and a more aggressive disease

Genetic and phenotypic attributes of splenic marginal zone lymphoma

: Splenic marginal zone B-cell lymphoma (SMZL) is a heterogeneous clinico-biological entity. The clinical course is variable, multiple genes are mutated with no unifying mechanism, and essential regulatory pathways and surrounding microenvironments are diverse. We sought to clarify the heterogeneity of SMZL by resolving different subgroups and their underlying genomic abnormalities, pathway signatures, and microenvironment compositions to uncover biomarkers and therapeutic vulnerabilities. We studied 303 SMZL spleen samples collected through the IELSG46 multicenter international study (NCT02945319) by using a multiplatform approach. We carried out genetic and phenotypic analyses, defined self-organized signatures, validated the findings in independent primary tumor metadata and in genetically modified mouse models, and determined correlations with outcome data. We identified 2 prominent genetic clusters in SMZL, termed NNK (58% of cases, harboring NF-κB, NOTCH, and KLF2 modules) and DMT (32% of cases, with DNA-damage response, MAPK, and TLR modules). Genetic aberrations in multiple genes as well as cytogenetic and immunogenetic features distinguished NNK- from DMT-SMZLs. These genetic clusters not only have distinct underpinning biology, as judged by differences in gene-expression signatures, but also different outcomes, with inferior survival in NNK-SMZLs. Digital cytometry and in situ profiling segregated 2 basic types of SMZL immune microenvironments termed immune-suppressive SMZL (50% of cases, associated with inflammatory cells and immune checkpoint activation) and immune-silent SMZL (50% of cases, associated with an immune-excluded phenotype) with distinct mutational and clinical connotations. In summary, we propose a nosology of SMZL that can implement its classification and also aid in the development of rationally targeted treatments