The SMURF2-YY1-C-MYC Axis in the Germinal Center Reaction and Diffuse Large B Cell Lymphoma: A Dissertation

Diffuse large B cell lymphoma (DLBCL) is the most common non-Hodgkin’s lymphoma. Patients who fail conventional therapy (~50%) have a poor prognosis and few treatment options. It is essential to understand the underlying biological processes, the progression of the disease, and utilize this information to develop new therapeutics. DLBCL patients with high C-MYC expression have a poor prognosis and new therapeutics for these patients are needed. This thesis describes work testing the hypothesis that JQ1, which can indirectly inhibit C-MYC in some tumors, can be used as an effective treatment for DLBCL. Some tumors have an unknown mechanism causing high C-MYC expression, leading me to investigate the underlying mechanisms. YY1 is a transcriptional regulator of c- Myc and has been implicated in DLBCL and as a potential regulator of the germinal center (GC) reaction. DLBCL arises from GC cells or post-GC cells. I tested the hypothesis that YY1 regulates the GC reaction. SMURF2 is an E3-ubiquitin ligase for YY1 and a tumor suppressor for DLBCL. I was interested in examining the mechanism underlying the suppression of DLBCL by SMURF2 leading to the hypothesis that SMURF2 regulates the GC. This thesis shows JQ1 leads to cell death and cellular senescence in human DLBCL cells. I conclude that BRD4 inhibition by JQ1 or derivatives could provide a new therapeutic avenue for DLBCL patients. I also show loss of YY1 perturbs the GC by decreasing the dark zone and increasing apoptosis. Finally I show modulation of SMURF2 does not affect the GC, suggesting SMURF2 utilizes a different mechanism to act as a tumor suppressor and may not modulate YY1 in the context of the GC

Smurf2 regulates hematopoietic stem cell self-renewal and aging

The age-dependent decline in the self-renewal capacity of stem cells plays a critical role in aging, but the precise mechanisms underlying this decline are not well understood. By limiting proliferative capacity, senescence is thought to play an important role in age-dependent decline of stem cell self-renewal, although direct evidence supporting this hypothesis is largely lacking. We have previously identified the E3 ubiquitin ligase Smurf2 as a critical regulator of senescence. In this study, we found that mice deficient in Smurf2 had an expanded hematopoietic stem cell (HSC) compartment in bone marrow under normal homeostatic conditions, and this expansion was associated with enhanced proliferation and reduced quiescence of HSCs. Surprisingly, increased cycling and reduced quiescence of HSCs in Smurf2-deficient mice did not lead to premature exhaustion of stem cells. Instead, HSCs in aged Smurf2-deficient mice had a significantly better repopulating capacity than aged wild-type HSCs, suggesting that decline in HSC function with age is Smurf2 dependent. Furthermore, Smurf2-deficient HSCs exhibited elevated long-term self-renewal capacity and diminished exhaustion in serial transplantation. As we found that the expression of Smurf2 was increased with age and in response to regenerative stress during serial transplantation, our findings suggest that Smurf2 plays an important role in regulating HSC self-renewal and aging

Inhibition of Bromodomain Proteins in Treatment of Diffuse Large B-cell Lymphoma

Only ~50% of patients with diffuse large B-cell lymphoma (DLBCL), the most common and aggressive subtype of non-Hodgkin’s lymphoma, enter long-term remission after standard chemotherapy, and patients who do not respond to treatment have few options. Therefore, there is a critical need for effective and targeted therapeutics for DLBCL. Recent studies highlight the incidence of increased c-MYC protein in DLBCL and the correlation between high levels of c-MYC and poor survival prognosis of DLBCL patients, suggesting that c-MYC is a compelling therapeutic target for DLBCL therapy. The small molecule JQ1 suppresses c-MYC expression through inhibition of the BET family of bromodomain proteins. We show that JQ1 efficiently inhibited cell proliferation of human DLBCL cells regardless of their molecular subtypes, suggesting a broad effect of JQ1 in DLBCL. After JQ1 treatment, initial G1 arrest in DLBCL cells was followed by either apoptosis or senescence. In DLBCL cells treated with JQ1, we found that c-MYC expression was suppressed in the context of the natural, chromosomally-translocated or an amplified gene locus. Furthermore, JQ1 treatment significantly suppressed growth of DLBCL cells engrafted subcutaneously and improved survival of mice engrafted with DLBCL cells intraperitoneally. These results demonstrate that inhibition of the BET family of bromodomain proteins, and consequently c-MYC, has the potential clinical utility in DLBCL treatment

How Many Bodies Are There? Unexpected Discoveries About the Subtle Body and Psychic Body

1. Patrizio E. Tressoldi[1][1][⇑][2] 2. Luciano Pederzoli[2][3] 3. Patrizio Caini[2][3] 4. Alessandro Ferrini[2][3] 5. Simone Melloni[2][3] 6. Elena Prati[2][3] 7. Diana Richeldi[2][3] 8. Florentina Richeldi[2][3] 9. Alice Trabucco[2][3] 1. 1Universita di Padova, Italy 2. 2EvanLab, Firenze, Italy 1. Patrizio E. Tressoldi, Universita di Padova, via Venezia 8, Padova 35131, Italy. Email: patrizio.tressoldi{at}unipd.it The possibility to induce real out-of-body experiences (OBEs) using hypnotic inductions, with the opportunity to interview participants during their experience, permits to investigate in depth the characteristics of different aspects of this particular state of consciousness from a first-person point of view. In this article, six selected participants report the description of another "body" we named "subtle body," identified as an intermediate entity between the physical body (Pb) and their "Self" or "I-identity" that was named "psychic body," and their relationships and characteristics. The "subtle body" was described as a sort of white silvered cloud surrounding the Pb, with a particular enlargement of its hands and feet that could move quickly like flying from one place to another even if less easily than the "psychic body," and a vague sense of attrition was perceived when passing through walls. Similar to the "psychic body," the "subtle" one too could move forward and backward in time even if they did not seem perceiving the sense of time. The "subtle body" was referred to be connected with the physical one by a sort of white brilliant link sometimes described like a silvered string more or less visible, whereas no visible links were identified between the "subtle body" and the "psychic" one. These reports were compared with similar descriptions deriving from the Vedanta philosophy and Theosophical tradition. [1]: #aff-1 [2]: #corresp-1 [3]: #aff-

Stiffness and slip in multi-dowel flitch-plate timber connections

Large multi-dowel connections can provide the strong and ductile connections required for large, highly-loaded timber structures, but their slip under load is not well understood. This is important because accumulated local displacements at connections can have significant implications for overall building serviceability. Empirical relationships for the slip of a single-dowel connection do not capture the dowel interaction effects of the multi-dowel connections used in larger structures. We present the results of an experimental test series and probabilistic numerical analysis investigating the development of stiffness in multi-dowel timber flitch plate connections. The influence of the diameter and number of dowels on the stiffness of the connection are investigated, including the influence of off-centring of dowels due to manufacturing tolerances. The test series is used to validate a probabilistic model for the stiffness of such a connection. The model incorporates the nonlinear stiffness and hole opening observed in single-dowel connections to predict the behaviour of the group. The study shows that the random off-centring of dowels in multi-dowel connections reduces the range of displacements over which the connection displays zero stiffness, but that this zone is not eliminated as a result of irreversible hole opening under load

Medical competence, anatomy and the polity in seventeenth-century Rome

At the centre of this article are two physicians active in Rome between 1600 and 1630 who combined medical practice with broader involvement in the dynamic cultural, economic and political scene of the centre of the Catholic world. The city's distinctive and very influential social landscape magnified issues of career-building and allows us to recapture physicians’ different strategies of self-fashioning at a time of major social and religious reorganization. At one level, reconstructing Johannes Faber and Giulio Mancini's medical education, arrival in Rome and overlapping but different career trajectories contributes to research on physicians’ identity in early modern Italian states. Most remarkable are their access to different segments of Roman society, including a dynamic art market, and their diplomatic and political role, claimed as well as real. But following these physicians from hospitals to courts, including that of the Pope, and from tribunals to the university and analysing the wide range of their writing – from medico-legal consilia to political essays and reports of anatomical investigations – also enriches our view of medical practice, which included, but went beyond, the bedside. Furthermore, their activities demand that we reassess the complex place of anatomical investigations in a courtly society, and start recovering the fundamental role played by hospitals – those quintessential Catholic institutions – as sites of routine dissections for both medical teaching and research. (pp. 551–567

Increased placental expression of cannabinoid receptor 1 in preeclampsia

BackgroundThe endocannabinoid system plays a key role in female reproduction, including implantation, decidualization and placentation. In the present study, we aimed to analyze cannabinoid receptor 1 (CB1), CB2 and fatty acid amid hydrolase (FAAH) expressions and localization in normal and preeclamptic placenta, in order to determine whether placental endocannabinoid expression pattern differs between normal pregnancy and preeclampsia.MethodsEighteen preeclamptic patients and 18 normotensive, healthy pregnant women with uncomplicated pregnancies were involved in our case inverted question markcontrol study. We determined CB1, CB2 and FAAH expressions by Western blotting and immunohistochemistry in placental samples collected directly after Cesarean section.ResultsCB1 expression semi-quantified by Western blotting was significantly higher in preeclamptic placenta, and these findings were confirmed by immunohistochemistry. CB1 immunoreactivity was markedly stronger in syncytiotrophoblasts, the mesenchymal core, decidua, villous capillary endothelial and smooth muscle cells, as well as in the amnion in preeclamptic samples compared to normal pregnancies. However, we did not find significant differences between preeclamptic and normal placenta in terms of CB2 and FAAH expressions and immunoreactivity.ConclusionsWe observed markedly higher expression of CB1 protein in preeclamptic placental tissue. Increased CB1 expression might cause abnormal decidualization and impair trophoblast invasion, thus being involved in the pathogenesis of preeclampsia. Nevertheless, we did not find significant differences between preeclamptic and normal placental tissue regarding CB2 and FAAH expressions. While the detailed pathogenesis of preeclampsia is still unclear, the endocannabinoid system could play a role in the development of the disease

Linguistic profile automated characterisation in pluripotential clinical high-risk mental state (CHARMS) conditions: methodology of a multicentre observational study

Introduction: Language is usually considered the social vehicle of thought in intersubjective communications. However, the relationship between language and high- order cognition seems to evade this canonical and unidirectional description (ie, the notion of language as a simple means of thought communication). In recent years, clinical high at-risk mental state (CHARMS) criteria (evolved from the Ultra-High-Risk paradigm) and the introduction of the Clinical Staging system have been proposed to address the dynamicity of early psychopathology. At the same time, natural language processing (NLP) techniques have greatly evolved and have been successfully applied to investigate different neuropsychiatric conditions. The combination of at-risk mental state paradigm, clinical staging system and automated NLP methods, the latter applied on spoken language transcripts, could represent a useful and convenient approach to the problem of early psychopathological distress within a transdiagnostic risk paradigm. Methods and analysis: Help-seeking young people presenting psychological distress (CHARMS+/− and Clinical Stage 1a or 1b; target sample size for both groups n=90) will be assessed through several psychometric tools and multiple speech analyses during an observational period of 1-year, in the context of an Italian multicentric study. Subjects will be enrolled in different contexts: Department of Neuroscience, Rehabilitation, Ophthalmology, Genetics, Maternal and Child Health (DINOGMI), Section of Psychiatry, University of Genoa—IRCCS Ospedale Policlinico San Martino, Genoa, Italy; Mental Health Department—territorial mental services (ASL 3—Genoa), Genoa, Italy; and Mental Health Department—territorial mental services (AUSL—Piacenza), Piacenza, Italy. The conversion rate to full-blown psychopathology (CS 2) will be evaluated over 2 years of clinical observation, to further confirm the predictive and discriminative value of CHARMS criteria and to verify the possibility of enriching them with several linguistic features, derived from a fine-grained automated linguistic analysis of speech. Ethics and dissemination: The methodology described in this study adheres to ethical principles as formulated in the Declaration of Helsinki and is compatible with International Conference on Harmonization (ICH)-good clinical practice. The research protocol was reviewed and approved by two different ethics committees (CER Liguria approval code: 591/2020—id.10993; Comitato Etico dell’Area Vasta Emilia Nord approval code: 2022/0071963). Participants will provide their written informed consent prior to study enrolment and parental consent will be needed in the case of participants aged less than 18 years old. Experimental results will be carefully shared through publication in peer- reviewed journals, to ensure proper data reproducibility. Trial registration number DOI:10.17605/OSF.IO/BQZTN

Notulae to the Italian flora of algae, bryophytes, fungi and lichens: 8

In this contribution, new data concerning algae, bryophytes, fungi, and lichens of the Italian flora are presented. It includes new records and confirmations for the algae genus Chara, the bryophyte genera Homalia, Mannia, and Tortella, the fungal genera Cortinarius, Russula, and Stereum, and the lichen genera Cetrelia, Cladonia, Enterographa, Graphis, Lecanora, Lepraria, Multiclavula, Mycomicrothelia, Parmelia, Peltigera, Pleopsidium, Psora, Scytinium, Umbilicaria, and Rhizocarpon