1,486 research outputs found
Error propagation in reprocessing input volume measurement. A study based on experiments and computer simulation
No full textDetermination of Material Unaccounted For (MUF) in Input verification of reprocessing plant
Anche il carcere è un luogo della città
No full textLo spazio del carcere è spesso considerato, nella città come nell’immaginario collettivo, un luogo lontano, da nascondere, circondato da una sorta di sospetto difficile da rimuovere. Maturato a partire dalla conduzione di un Laboratorio di progettazione sul carcere di Bollate (Milano), svolto presso il Politecnico di Milano, il saggio si interroga sul posto e sul ruolo che questo luogo così particolare può assumere nella città. L’ipotesi è che anch’esso debba essere considerato un luogo urbano, e che un suo ripensamento debba coinvolgere molteplici dimensioni: le modalità di accedervi, la definizione di spazi di soglia tra interno ed esterno come dotazioni collettive per la città, la natura complessa del recinto inteso non solo come elemento di separazione, l’esplorazione di forme di abitabilità per certi versi estreme, la possibile articolazione dello spazio aperto compreso dentro le mura. Pensare al carcere come “spazio abitabile” è il tema generale che naturalmente attraversa sia il saggio, sia le esplorazioni progettuali da cui esso trae spunto
A phospho-specific flow cytometry study reveals the impact of BCR signaling activation in mantle cell lymphoma progression and therapy
No full textMantle cell lymphoma (MCL) is a rare and heterogeneous subtype of Non-Hodgkin lymphoma characterized by the expansion of mature
B-cells in the mantle zone of the lymph nodes. Recent evidence supports that B-cell receptor (BCR) signaling is pivotal for MCL initiation
and progression and is a target for therapeutic treatment. However, drug resistance inevitably emerges.
In this study, we characterized the activation status of phosphoproteins on the BCR pathway, and evaluated the impact of the BCR in
MCL progression and response to therapy.
We measured the activation status of 9 BCR signaling kinases (SYK, LCK, BTK, PLCγ2, p38, ERK1/2, AKT, NF-κB p65, STAT5) in
peripheral blood mononuclear cells (PBMC) from 30 MCL patients and 10 healthy donors, in the basal and anti-IgM modulated conditions,
at single cell level using phospho-specific flow cytometry.
BCR modulation with anti-IgM determined a heterogeneous activation of the BCR signaling among patients’ samples, with identification
of two clusters showing differential responses to BCR stimulation. The cluster comprising samples with higher BCR signaling response
(HR) was associated with shorter survival than samples grouped in the lower BCR signaling response (LR) cluster. This finding was
further corroborated with time-to-event analyses, which showed that the MCL international prognostic index (MIPI) high-risk category
together with high STAT5 response to the stimulation were significant predictors of shorter PFS and OS. In addition, MIPI high-risk
category combined with high SYK response predicted shorter OS. While basal BCR activation did not provide prognostic information,
higher constitutive activation of AKT was predictive of inferior response to the Bruton's tyrosine kinase (BTK) inhibitor ibrutinib.
In conclusion, we identified BCR signaling activation profiles that were associated with poorer clinical outcome and resistance to ibrutinib.
This study highlighted the prognostic and predictive significance of BCR activity in MCL and advanced our understanding of signaling
heterogeneity underlying clinical behavior of MCL. A future challenge is the integration of BCR signaling data with genetic signature to
predict patient’s clinical behavior and drug response for a more personalized treatment approach. We thank Fondazione Italiana Linfomi
(PGR Ed. 2019) for funding support
SARA: A Montecarlo simulation code for NRTA implementation to EUREX pilot reprocessing plant
No full textImplementation of Near Real Time Accountancy procedures for Eurex Reprocessing Plan
PROGNOSTIC AND PREDICTIVE SIGNIFICANCE OF THE B-CELL RECEPTOR SIGNALING IN MANTLE CELL LYMPHOMA
No full textMantle cell lymphoma (MCL) is a rare subtype of Non-Hodgkin
lymphoma characterized by the expansion of mature B-cells in the
mantle zone of the lymph nodes. MCL presents a high clinical vari-
ability, with some patients experiencing an indolent disease while
others characterized by an aggressive clinical course. Recent evi-
dence supports that B-cell receptor (BCR) signaling is crucial for
MCL initiation and progression and is a target for therapeutic inter-
vention. However, drug resistance inevitably emerges. Therefore, the
definition of parameters identifying high-risk patients for aggressive
disease and therapy resistance is an unmet need in MCL management.
We measured the activation status of 9 BCR signaling kinases (SYK,
LCK, BTK, PLCγ2, p38, ERK1/2, AKT, NF-κB p65, STAT5) in
peripheral blood mononuclear cells (PBMC) from 30 MCL patients
and 10 healthy donors, in the basal and anti-IgM modulated condi-
tions, using phospho-specific flow cytometry. To measure phospho-
rylation statuses as well as responses to external stimulation of the
signaling proteins, flow cytometry data were normalized with respect
to the controls and subjected to unsupervised hierarchical clustering
analysis (HCA). Progression free survival (PFS) and overall survival
(OS) curves were estimated using the Kaplan-Meier method and
compared using the log-rank test. Univariate and bivariate models
for PFS and OS were performed using Cox proportional hazard
regression. BCR modulation with anti-IgM determined a heteroge-
neous activation of the BCR signaling among patients’ samples, with
identification of two clusters showing differential responses to BCR
stimulation. The cluster comprising samples with higher BCR sig-
naling response (HR) was associated with shorter survival than sam-
ples grouped in the lower BCR signaling response (LR) cluster (p=0.
042 and p=0. 041 for PFS and OS, respectively). This finding was
confirmed with time-to-event analyses, which showed that the MCL
international prognostic index (MIPI) high-risk category together
with high STAT5 response to the stimulation were significant pre-
dictors of shorter PFS and OS. In addition, MIPI high-risk category
combined with high SYK response predicted shorter OS. While basal
BCR activation did not provide prognostic information, higher con-
stitutive activation of AKT was predictive of inferior response to the
Bruton’s tyrosine kinase inhibitor (BTK[/i] ibrutinib. In conclusion,
we identified BCR signaling activation profiles that were associated
with poorer clinical outcome and resistance to ibrutinib. This study
highlighted the prognostic and predictive significance of BCR activ-
ity in MCL and advanced our understanding of signaling heterogene-
ity underlying clinical behavior of MCL. A future challenge is the
integration of BCR signaling data with genetic signature to predict
patient’s clinical behavior and drug response for a more personalized treatment approach. We thank Fondazione Italiana Linfomi (PGR
Ed. 2019) for funding support
Mathematics in a fun and playful way : A video –observation study of children`s play in preschool
No full textAbstract Swedish title; Matematik på ett lekfullt sätt – En videoobservationsstudie av barns lek i förskolan. English title; Mathematics in a fun and playful way. A video –observation study of children`s play in preschool Author; Barbra Aparo Language; Swedish with an English summary. Supervisor; Gabriella Gejard Examiner; Pia Maria Ivarsson Term; Spring 2017 The purpose of this study is to discover what types of mathematical activities that can arise from children`s free play. The study focuses on children between the ages of 3 and 6 years. In this paper, i explore how children through play engage in different mathematical activities. The research questions are; In what kind of situations do children use mathematical concepts? How can the mathematics that arise in children`s play be categorized? The study is based on Alan Bishops six cultural and historic mathematical activities and variations theory. Alan Bishops mathematical activities were used to analyse and categorise the different types of mathematics that children engage in during their free play. Variations theory was used as a method to describe how children learn those mathematical activities that arise in their play and what they need in order for learning to occur. Data were collected through qualitative methods and the primary data source for the study were video recordings. Results show that all the Alan Bishops mathematical activities arise in children`s free play. These activities are; counting, locating, measuring, designing, playing and explaining. The results also show that children in their free play explore patterns and shape, compare sizes, measurement, weight and volume, develop problem-solving, count things, group, sort, categorize and classify various objects. Results also show how children in various play situations used mathematical concepts like large –small, heavy –light, long –short, same –different, more –less etc. Keywords: Free play, play, mathematics, variation theory, children, mathematical activities, preschool Sammanfattning Syftet med studien är att genom observationer undersöka om och i så fall vilken matematik som förekommer i barns fria lek. Jag har undersökt och observerat hur barn genom lek engagerar sig i olika matematiska aktiviteter. Denna uppsats är begränsad till att endast undersöka barn mellan 3 och 6 år. Jag använder mig av följande frågeställningar; I vilka situationer/lekar använder sig barnen av matematiska begrepp? Hur kan matematiken som förekommer i barnens lekar kategoriseras? De teoretiska utgångspunkterna är Alan Bishops sex kulturella och historiska matematiska aktiviteter och variationsteori. Bishops sex matematiska aktiviteter har jag valt för att analysera och kategorisera de olika matematiska aktiviteter som förekommer i barns fria lek. Jag har använt mig av variationsteori som ett verktyg för att beskriva hur barn lär matematik i leken, och vad de behöver för att kunna lära sig. I studien har jag använt mig av en kvalitativ metod i form av videoobservationer. Resultatet visar att alla Bishops matematiska aktiviteter förekommer i barns fria lek. Dessa kategorier är räkna, mäta, lokalisera, förklara, konstruera/designa och leka. Resultatet visar även att barn i den fria leken får möjlighet att utforska mönster och former, storlek, volym, mäta, räkna, utveckla problemlösning, sortera och kategorisera mängder av olika föremål. Resultatet visar också att barn i olika leksituationer bekantar sig med begreppen lika –olika, lång –kort, smal –bred, hög –låg, och så vidare. Nyckelord: Fri lek, lek, matematik, matematiska aktiviteter, variationsteori, barn, förskola
ACTIVATION OF BCR SIGNALING REWIRES THE RESPONSE OF BCR-ASSOCIATED KINASES TO IBRUTINIB IN MANTLE CELL LYMPHOMA CELLS
No full textMantle cell lymphoma (MCL) is a rare B-cell malignancy with sig-
nificant clinical heterogeneity. We have recently shown that a deeper re-
sponse of MCL cells to B-cell receptor (BCR) stimulation identified a
subset of patients with a higher risk of progression. Although therapies
targeting the BCR, such as the BTK inhibitor ibrutinib (Ibr), have a high
initial response rate in MCL, relapse remains a challenge. To investigate
the ability of Ibr to modulate the BCR signaling network, we analyzed
the activating phosphorylation status of BCR-associated kinases (BAK),
i.e., SYK, PLCγ2, STAT5, ERK1/2, NF-κB p65, AKT, BTK, STAT3, in
cells from peripheral blood of 29 MCL patients at diagnosis following Ibr treatment in vitro, in the basal condition and after BCR stimulation
with anti-IgM, using phospho-specific flow cytometry, a multiparametric
assay allowing functional signaling analysis at a single-cell level, com-
bined with fluorescent cell barcoding. In the basal condition, Ibr induced
a significant average reduction of phosphorylation level for all BAKs
but NF-κB p65 and STAT3. In the BCR-stimulated condition, we de-
tected a significant average reduction of phosphorylation for all BAKs,
including NF-κB p65, but STAT3. Comparison of the phosphorylation
responses to Ibr for each BAK between the basal and BCR-stimulated
conditions showed that the phosphorylation reduction in response to Ibr
was significantly deeper in the BCR-stimulated condition compared with
the basal one for SYK, ERK1/2, NF-κB p65, and AKT. In contrast, re-
duction in BTK phosphorylation resulted attenuated, although not sig-
nificantly, in the BCR-stimulated condition compared with the basal one
(Figure 1). In conclusion, our data show that in the basal condition most
BAKs are highly connected in response to Ibr, thus suggesting that these
proteins belong to the same signaling subnetwork. Importantly, the acti-
vation of the BCR quantitatively and qualitatively rewires the network
of BAKs sensible to Ibr, thus suggesting that the microenvironment may
influence the response of tumor cells to therapies. Moreover, identifying
novel BAKs that are inhibited by Ibr may form the rationale to target
multiple signaling nodes to overcome resistance in MCL.
Thank to Excellence Project 2023-2027 of the DNBM, University of
Verona funded by MUR; European Union, MUR and MSAL (PNRR-
PNC-E3-2022-23683266 PNC-HLS-DA-INNOVA and PNRR-TR1-2023-
12378287) for funding support
THE PROGNOSTIC ROLE OF DEL 9P21.3 (CDKN2A) IN PATIENTS WITH MANTLE CELL LYMPHOMA. RESULTS FROM THE FIL_MANTLE-FIRST BIO STUDY
No full textMantle Cell Lymphoma (MCL) is an aggressive malignancy with
variable clinical behaviour, largely reflecting its molecular heterogeneity.
The genomic landscape of MCL encompasses gene mutations with
strong prognostic implications (i.e., TP53, NOTCH1/2, KMT2D, etc),and secondary genetic events such as copy number variations (CNVs),
which are also implicated in pathogenesis and prognosis. We analyzed
genomic DNA (gDNA) extracted from diagnostic formalin-fixed paraf-
fin-embedded (FFPE) lymph nodes or extra-nodal biopsies of 81 treat-
ment-naïve patients with MCL enrolled in the MANTLE-FIRST BIO
study (NCT04882475). We performed targeted sequencing on the Illu-
mina platform using a customized panel. The somatic mutations (SNVs)
were annotated with MuTect2, while CNVs with CNVkit. To be in-
cluded, patients had to have experienced relapse or progression (R/R)
after induction therapy and during the observation period, which was
held by centres from the Fondazione Italiana Linfomi (FIL). Results were
analyzed according to time to first R/R and divided following accepted
definition of early versus late progression of disease (POD). Overall, 18
CNVs were identified (11 amplifications and 7 deletions). The oncoprint
showed that Del13q14 (RB1, 34%,p=0.02), Del6q (21%, p=0.01), and
Del9p21.3 (CDKN2A, 19%, p=0.02) were significantly associated with
more frequent early-POD (Figure 1). CNVs’ unbiased clusterization
highlighted four different molecular clusters, which could split the cohort
into groups with significantly different time to first POD (p=0.01).
Specifically, cluster1 was enriched by Del 9p21.3 (CDKN2A). A back-
ward multivariate analysis was performed in terms of time to first POD,
including all variables that were significant in univariate analysis, as
tumor morphology (classic vs blastoid/pleomorphic), MIPI score (high
versus others), Ki-67 (≤30% vs >30%), Del 9p21.3 (CDKN2A), Del
13q14 (RB1), mutations of TP53, EZH2, BTK, NFKBIE, and STAT3.
Del 9p21.3 (CDKN2A) emerged as the only significant alteration asso-
ciated with significantly inferior time to first POD (p=0.01, HR=2.47),
together with blastoid/pleomorphic morphology (p=0.14, HR 1.74), and
high-risk MIPI (p<0.001, HR 3.72). Our comprehensive analysis shows
that CNVs reflect the high heterogeneity of MCL genomic landscape,
and reveal a strong and independent association between Del 9p21.3
(CDKN2A) and the occurrence of early POD. We thank FIL for funding
support (Premio Giovani Ricercatori, Ed. 2019)
GENOMIC ANALYSES OF PATIENTS WITH MANTLE CELL LYMPHOMA THAT WERE REFRACTORY OR RELAPSED AFTER INDUCTION THERAPY: RESULTS FROM THE FIL_MANTLE- FIRST BIO STUDY
No full textMANTLE-FIRST BIO retrospective study enrolls mantle cell lym-
phoma (MCL) patients, aged 18-80 years, experiencing first relapse or
refractoriness (R/R) to frontline chemo-immunotherapy (CIT). Patients
with available diagnostic samples other than bone marrow biopsy from
16 centers affiliated with Fondazione Italiana Linfomi (FIL) were en-
rolled in this study. We performed a targeted sequencing analysis (Illu-
mina platform) on genomic DNA extracted from formalin-fixed
paraffin-embedded diagnostic samples (lymph node or extra-nodal tis-
sue). A customized panel including 37 genes was used to identify gene
mutations associated with different time to first progression of disease
(time to POD). Single nucleotide variants (SNVs) calling was performed
with MuTect2. The previously defined threshold of 24 months since
MCL diagnosis was used to define patients as early- or late-POD. The
primary endpoint was to investigate the correlation between time to POD
and clinical or molecular features. Eighty-one MCL patients (29 early-
and 52 late-POD) were included. Median age was 67 years, 62 were
males, median time to POD was 35.3 months (range 3-149.2), and me-
dian follow-up from lymphoma diagnosis was 69.9 months. Data anal-
ysis identified ATM as the most frequently mutated gene (42%), followed
by NOTCH1 (32%), SMARCA4 (30%), TP53 (29%) and KMT2D (26%)
(Figure 1). Among mutated genes, TP53 (p=0.0003, HR 2.21), NFKBIE
(p=0.01, HR 2.97) and STAT3 (p=0.02, HR 2.78) were significantly as-
sociated with shorter time to POD at univariate analysis. Non-negative
matrix factorization identified four different clusters with well-defined
molecular composition and with significantly different time to POD
(p<0.011). Cluster 1 included patients who presented SNVs such as
NOTCH1 (40%), TP53 (38%), and KMT2D (20%) that were associated
with the shortest time to POD. In multivariate analysis, TP53 (p=0.01,
HR 2.15), tumour morphology (classic versus blastoid/pleomorphic,
p=0.14, HR 1.74) and MIPI score (high versus others, p<0.001, HR 3.72)
were independently associated with shorter time to POD. In conclusion,
our MCL cohort showed high genetic complexity that underlies the het-
erogeneous clinical behavior. Our findings may contribute to the identi-
fication of distinct biological profiles that are associated to significantly
different time to POD.
We thank FIL for funding support (Premio Giovani Ricercatori, Ed.
2019)
CATALASE BLOCKING INDUCES APOPTOSIS AND POTENTIATES VENETOCLAX-INDUCED CELL DEATH IN CHRONIC LYMPHOCYTIC LEUKEMIA CELLS
No full textChronic lymphocytic leukemia (CLL) is an incurable disease char-
acterized by a highly variable clinical behavior. Although targeted ther-
apies, such as the BCL-2 inhibitor Venetoclax, have high initial response
rates, relapse remains a challenge. We have recently shown that low lev-
els of the catalase antioxidant enzyme identify an indolent CLL, leading
us to hypothesize that catalase downregulation could induce escalated
levels of reactive oxygen species (ROS) that promote antitumor signals
such as cell death, thus accounting for less aggressive behavior and
higher therapy sensitivity of cancer cells. To verify our hypothesis, we
manipulated catalase activity using the specific inhibitor 3-amino-1,2,4-
triazole (ATZ) in MEC1 CLL cell line and primary cells from CLL pa-
tients. Moreover, in MEC1 cells we targeted catalase using siRNA
technology. We analyzed ROS levels using CM-H2DCFDA fluorescent
probe while we measured apoptosis with PI-Annexin V staining and flow
cytometry. First, we observed that in vitro spontaneous apoptosis of pri-
mary CLL cells was associated with a significant increased accumulation
of ROS (Pearson r=0.7552; p=0.0186), thus suggesting a functional role
of ROS in inducing apoptosis in CLL cells. Importantly, blocking cata-
lase either with ATZ or siRNA induced escalated ROS levels and in-
creased apoptosis in leukemic cells (Figure 1A). To extend our findings
to a therapeutic setting, we analyzed the ability of catalase to modulate
cell survival in vitro in the presence of Venetoclax. Remarkably, inhibit-
ing catalase with ATZ in combination with Venetoclax significantly po-
tentiated apoptosis in CLL cells compared to those induced by each
single agent (Figure 1B). These data support the hypothesis that catalase
might play a critical role in regulating susceptibility to spontaneous as
well as BCL-2 inhibitor-induced apoptosis in CLL, thus influencing the
behavior of cancer cells and contributing to resistance. Moreover, this
study provides the rationale for developing novel precision therapies tar-
geting redox pathways that, alone or in combination with currently used
therapies, could overcome resistance in CLL and other B-cell malignan-
cies where BCL-2 inhibitor is used, such as mantle cells lymphoma. We
thank the Excellence Project 2023-2027 of the DNBM of the University
of Verona funded by MUR; European Union, MUR and MSAL (PNRR
PNC-E3-2022-23683266 PNC-HLS-DA, INNOVA and PNRR-TR1-
2023-12378287) for funding support
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