Loss of the candidate tumor suppressor ZEB1 (TCF8, ZFHX1A) in Sézary syndrome

Cutaneous T-cell lymphoma is a group of incurable extranodal non-Hodgkin lymphomas that develop from the skin-homing CD4+ T cell. Mycosis fungoides and Sézary syndrome are the most common histological subtypes. Although next-generation sequencing data provided significant advances in the comprehension of the genetic basis of this lymphoma, there is not uniform consensus on the identity and prevalence of putative driver genes for this heterogeneous group of tumors. Additional studies may increase the knowledge about the complex genetic etiology characterizing this lymphoma. We used SNP6 arrays and GISTIC algorithm to prioritize a list of focal somatic copy-number alterations in a dataset of multiple sequential samples from 21 Sézary syndrome patients. Our results confirmed a prevalence of significant focal deletions over amplifications: single well-known tumor suppressors, such as TP53, PTEN, and RB1, are targeted by these aberrations. In our cohort, ZEB1 (TCF8, ZFHX1A) spans a deletion having the highest level of significance. In a larger group of 43 patients, we found that ZEB1 is affected by deletions and somatic inactivating mutations in 46.5% of cases; also, we found potentially relevant ZEB1 germline variants. The survival analysis shows a worse clinical course for patients with ZEB1 biallelic inactivation. Multiple abnormal expression signatures were found associated with ZEB1 depletion in Sézary patients we verified that ZEB1 exerts a role in oxidative response of Sézary cells. Our data confirm the importance of deletions in the pathogenesis of cutaneous T-cell lymphoma. The characterization of ZEB1 abnormalities in Sézary syndrome fulfils the criteria of a canonical tumor suppressor gene. Although additional confirmations are needed, our findings suggest, for the first time, that ZEB1 germline variants might contribute to the risk of developing this disease. Also, we provide evidence that ZEB1 activity in Sézary cells, influencing the reactive oxygen species production, affects cell viability and apoptosis

Purification, characterization and molecular cloning of the major chitinase from Tenebrio molitor larval midgut

Insect chitinases are involved in degradation of chitin from the exoskeleton cuticle or from midgut peritrophic membrane during molts. cDNAs coding for insect cuticular and gut chitinases were cloned, but only chitinases from moulting fluid were purified and characterized. In this study the major digestive chitinase from T. molitor midgut (TmChi) was purified to homogeneity, characterized and sequenced after cDNA cloning. TmChi is secreted by midgut epithelial cells, has a molecular weight of 44 kDa and is unstable in the presence of midgut proteinases. TmChi shows strong substrate inhibition when acting on umbelliferyl-derivatives of chitobio- and chitotriosaccharides, but has normal Michaelis kinetics with the N-acetylglucosamine derivative as substrate. TmChi has very low activity against colloidal chitin, but effectively converts oligosaccharides to shorter fragments. The best substrate for TmChi is chitopentaose, with highest kcat/KM value. Sequence analysis and chemical modification experiments showed that the TmChi active site contains carboxylic groups and a tryptophane, which are known to be important for catalysis in family 18 chitinases. Modification with p-hidroximercuribenzoate of a cysteine residue, which is exposed after substrate binding, leads to complete inactivation of the enzyme. TmChi mRNA encodes a signal peptide plus a protein with 37 kDa and high similarity with other insect chitinases from family 18. Surprisingly, this gene does not encode the C-terminal Ser-Thr-rich connector and chitin-binding domain normally present in chitinases. The special features of TmChi probably result from its adaptation to digest chitin-rich food without damaging the peritrophic membrane. © 2006 Elsevier Ltd. All rights reserved

T Cell Leukemia/Lymphoma 1A is essential for mouse epidermal keratinocytes proliferation promoted by insulin-like growth factor 1

T Cell Leukemia/Lymphoma 1A is expressed during B-cell differentiation and, when overexpressed, acts as an oncogene in mouse (Tcl1a) and human (TCL1A) B-cell chronic lymphocytic leukemia (B-CLL) and T-cell prolymphocytic leukemia (T-PLL). Furthermore, in the murine system Tcl1a is expressed in the ovary, testis and in pre-implantation embryos, where it plays an important role in blastomere proliferation and in embryonic stem cell (ESC) proliferation and self-renewal. We have also observed that Tcl1-/-adult mice exhibit alopecia and deep ulcerations. This finding has led us to investigate the role of TCL1 in mouse skin and hair follicles. We have found that TCL1 is expressed in the proliferative structure (i.e.The secondary hair germ) and in the stem cell niche (i.e.The bulge) of the hair follicle during regeneration phase and it is constitutively expressed in the basal layer of epidermis where it is required for the correct proliferative-differentiation program of the keratinocytes (KCs). Taking advantage of the murine models we have generated, including the Tcl1-/-and the K14-TCL1 transgenic mouse, we have analysed the function of TCL1 in mouse KCs and the molecular pathways involved. We provide evidence that in the epidermal compartment TCL1 has a role in the regulation of KC proliferation, differentiation, and apoptosis. In particular, the colony-forming efficiency (CFE) and the insulin-like growth factor 1 (IGF1)-induced proliferation are dramatically impaired, while apoptosis is increased, in KCs from Tcl1-/-mice when compared to WT. Moreover, the expression of differentiation markers such as cytokeratin 6 (KRT6), filaggrin (FLG) and involucrin (IVL) are profoundly altered in mutant mice (Tcl1-/-). Importantly, by over-expressing TCL1A in basal KCs of the K14-TCL1 transgenic mouse model, we observed a significant rescue of cell proliferation, differentiation and apoptosis of the mutant phenotype. Finally, we found TCL1 to act, at least in part, via increasing phospho-ERK1/2 and decreasing phospho-P38 MAPK. Hence, our data demonstrate that regulated levels of Tcl1a are necessary for the correct proliferation and differentiation of the interfollicular KC

Association of processed meat intake and obesity in a population-based study of Japanese-Brazilians

OBJECTIVE: The aim of this study was to investigate the association between the consumption of processed meat with overall, abdominal, and overall with abdominal obesity in a Japanese-Brazilian population, which is known to be at cardiometabolic risk. SUBJECTS AND METHODS: A total of 329 men and 443 women aged ≥ 30 years were evaluated in a cross-sectional population-based survey. Diagnosis of overall obesity and abdominal obesity were based on the World Health Organization (WHO) criteria for Asians. Food intake was assessed by a validated food frequency questionaire. RESULTS: In men, processed meat intake was positively associated with overall with abdominal obesity (OR 2.97; 95%CI 1.13-7.78) after adjustment. In women, only the red meat group was associated with overall with abdominal obesity after adjustment (OR 0.47, 95%CI 0.23-0.96). CONCLUSION: Our results showed that high intakes of processed meats were associated with overall with abdominal obesity in male Japanese-Brazilians, but not in females.OBJETIVO: Investigar a associação entre consumo de alimentos embutidos e obesidade generalizada, abdominal e generalizada com abdominal em nipo-brasileiros de Bauru, SP. SUJEITOS E MÉTODOS: Quatrocentos e quarenta e três mulheres e 329 homens nipo-brasileiros não miscigenados (≥ 30 anos) foram avaliados em estudo transversal de base populacional. Para o diagnóstico de obesidade, foram empregados os critérios da Organização Mundial da Saúde para asiáticos. A ingestão de alimentos foi avaliada por meio de questionário de frequência alimentar validado. A ingestão foi estratificada em terços para análise. RESULTADOS: Nos homens, a ingestão de colesterol e alimentos embutidos mostrou-se positivamente associada à obesidade generalizada com abdominal quando o primeiro terço de ingestão foi comparado ao último, após ajustes (OR 2,97; IC95% 1,13-7,78). Em mulheres, somente o grupo das carnes vermelhas associou-se à obesidade geral com abdominal após ajustes (OR 0,47; IC95% 0,23-0,96). CONCLUSÃO: Ingestão elevada de alimentos embutidos associou-se à obesidade generalizada com adiposidade abdominal em homens nipo-brasileiros, mas não em mulheres.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade de São Paulo Faculdade de Saúde Pública Departamento de NutriçãoUniversidade Federal de São Paulo (UNIFESP) Departamento de Medicina PreventivaUNIFESP, Depto. de Medicina PreventivaSciEL

Blood and skin-derived Sezary cells: differences in proliferation-index, activation of PI3K/AKT/mTORC1 pathway and its prognostic relevance

Sézary syndrome (SS) is a rare and aggressive variant of Cutaneous T-Cell Lymphoma characterized by neoplastic distribution mainly involving blood, skin, and lymph-node. Although a role of the skin microenvironment in SS pathogenesis has long been hypothesized, its function in vivo is poorly characterized. To deepen this aspect, here we compared skin to blood-derived SS cells concurrently obtained from SS patients highlighting a greater proliferation-index and a PI3K/AKT/mTORC1 pathway activation level, particularly of mTOR protein, in skin-derived-SS cells. We proved that SDF-1 and CCL21 chemokines, both overexpressed in SS tissues, induce mTORC1 signaling activation, cell proliferation and Ki67 up-regulation in a SS-derived cell line and primary-SS cells. In a cohort of 43 SS cases, we observed recurrent copy number variations (CNV) of members belonging to this cascade, namely: loss of LKB1 (48%), PTEN (39%) and PDCD4 (35%) and gains of P70S6K (30%). These alterations represent druggable targets unraveling new therapeutic treatments as metformin here evaluated in vitro. Moreover, CNV of PTEN, PDCD4, and P70S6K, evaluated individually or in combination, are associated with reduced survival of SS patients. These data shed light on effects in vivo of skin-SS cells interaction underlying the prognostic and therapeutic relevance of mTORC1 pathway in SS

Biowaiver Monographs for Immediate‐Release Solid Oral Dosage Forms: Codeine Phosphate

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/106898/1/jps23977.pd

Following the Science? Scientific Knowledge Use in Entrepreneurial Ecosystem Policymaking

The academic community is often considered a relevant actor in variouspolicymaking arenas. The literature on the use and impact of research in policymak-ing has pointed to several ways that knowledge can flow between the academic worldto policy (and vice-versa), even though gaps in understanding this relationship existfor this in certain areas, as is the case with Entrepreneurship Ecosystem policies. Inthis sense, this study aims to identify and analyze how research and scientific knowl-edge have been used by policy actors in policy documents and policymaking processesrelated to entrepreneurial ecosystems (EEs) in the European Union. The study focuseson the role of the Joint Research Centre (JRC), a cluster of 10 scientific areas that formthe EU Science Hub. We used an altimetry tool (Overton) to identify the use of re-search in policy documents. The results indicate the prevalence of apoliticalapproachunderpinned by strategic use of knowledge and expertise as well as strong expert con-centration and convergence effects. We discuss these results in terms of their implica-tions for EEs policy and research and identify avenues for future research.publishedVersio

Fenomenologia del concetto di élite e network analysis come metodo di information retrieval

According to Sartori, “Language is the sine-qua-non instrument of knowing”. Nevertheless, social sciences often adopt terms lacking clear and precise definitions. Adopting the method proposed in Sartori (1984), the present Master’s thesis consists of an explicatory study aiming to clarify the concept of “elites”. First, the essential works concerning elites are identified by analyzing a large citation network. Then, the thesis proposes a definition of elite able to encompass those of previous works. The new definition attempts to include the concept of elites within the broader framework of the resource-dependence theories (Pfeffer e Salancik, 1978) and purposive theories (Coleman, 2005)

Tecniche di biologia molecolare nello studio dei sistemi chemosensoriali

TECNICHE DI BIOLOGIA MOLECOLARE NELLO STUDIO DEI SISTEMI CHEMOSENSORIALI RIASSUNTO La caratterizzazione ultrastrutturale a partire dagli anni sessanta e in seguito recentemente quella immunoistologica ha permesso di identificare cellule specializzate che possiedono diverse analogie morfologiche e molecolari con le cellule gustative ma non sono organizzate in calici gustativi. Queste cellule chemosensoriali solitarie (CCS) sono molto diffuse oltre che nella cavità orale anche nelle vie aeree e nell’apparato digerente. L’identificazione di queste cellule singole o disposte in cluster ha portato a concettualizzare l’esistenza di un sistema chemosensoriale diffuso (DCS) di cui le cellule gustative organizzate nei taste buds nella cavità orale sono solo la parte più evidente. Le funzioni del DCS sembrano coinvolgere la secrezione, la risposta immunitaria innata, l’assorbimento ma altri ruoli sono in parte ancora speculativi, e inoltre non tutti i tipi di cellule sono stati caratterizzati, lasciando aperte molte questioni che questo studio ha in parte analizzato utilizzando tecniche di biologia molecolare. In particolare sono state indagate tramite RT-PCR e western blot le differenti capacità chemorecettoriali degli organi di origine endodermica ricercando l’espressione genica dei recettori implicati nella percezione del gusto. Inoltre sono state coadiuvate ricerche finalizzate a meglio caratterizzare le cellule recettoriali all’interno dei taste buds, che risultano possedere un ventaglio di pathway molecolari molto ampio, comprendente elementi tipici delle vie aeree e dell’apparato gastrointestinale. È stato analizzato se l’espressione degli stessi geni sia modificabile a livello intestinale da diversi tipi di diete. I dati raccolti indicano una differente localizzazione dei recettori del dolce e dell’amaro lungo le vie aeree che hanno un ruolo di modulazione del movimento ciliare e della secrezione. Mentre la presenza dei recettori del dolce non va in maggior profondità della trachea, alcuni recettori dell’amaro risultano espressi anche nei polmoni. Invece lungo l’apparato gastrointestinale i recettori dell’amaro sembrano essere espressi in maniera selettiva e differenziata. L’intestino risulta inoltre sensibile a diete prolungate sovra esprimendo o sotto regolando l’espressione di proteine coinvolte nell’assorbimento e nella secrezione. In particolare è stato approfondito il sensing intestinale verso gli acidi grassi mediante quantificazione tramite PCR real time. I risultati indicano che diete iperlipidiche croniche abbattono sia l’espressione di CD36, una proteina con alta affinità per gli acidi grassi presente nella membrana apicale degli enterociti, sia l’espressione dei meccanismi di segnalazione di “saziazione” indotta dai grassi. Si tratta di un segnale, che diversamente da altri ormoni intestinali ha la capacità di prolungare la latenza tra i pasti, viene generato principalmente dalla produzione dell’ormone lipidico oleoil-etanolamide (OEA). Questo ulteriore studio oltre a evidenziare un nuovo aspetto negativo delle diete iperlipidiche, trova l’esistenza di una correlazione tra i diversi geni indagati che suggerisce che CD36 funga come sensore intestinale degli acidi grassi e abbia ruoli regolatori sui meccanismi di produzione di OEAMOLECULAR BIOLOGY TECHNIQUES IN CHEMOSENSORY SYSTEMS STUDY ABSTRACT Since 1960s, the ultrastructural and more recently the immunohystological characterization have identified specialized cells with several morphological and molecular similarities with the taste cells, without being organized into taste buds. These solitary chemosensory cells (CCS) are widespread not only in the oral cavity but even in the respiratory and gastrointestinal tract. The identification of these cell, that can be individually arranged or clustered, has led to conceptualize the existence of a diffuse chemosensory system (DCS) where the taste receptor cells organized within taste buds in the oral cavity are only the most obvious. The functions appear to involve the secretion and absorption of the DCS, and the innate immune response, but other roles are still partly speculative as not all cell types have been characterized, leaving opened many questions that this study has partly analyzed using molecular biology techniques. In particular, the different chemoreceptorial capacities of endodermal origin organs were assessed by RT-PCR and western blot examining the gene expression of those receptors that are involved in taste perception. We also performed experiments to better characterize the receptor cells within taste buds, showing that they have a very wide range of molecular pathways including typical features of the respiratory and gastrointestinal tract. It was analyzed whether the expression of these genes is modified by different types of diets at intestinal level. The data indicate a different localization of the bitter and sweet receptors along the respiratory pathway that have a role in the modulation of ciliary movement and secretion. Whereas the sweet receptors cannot be found deeper than trachea, some of the bitter receptors are also expressed in the lungs. On the other hand, along the gastrointestinal tract the bitter receptors appear to be expressed in a selective and differentiated manner. The intestine is also sensitive to prolonged diets, overexpressing or downregulating the expression of proteins involved in absorption and secretion. In particular, we studied the intestinal fatty acid sensing by real time PCR utilize. The results indicate that chronic high-fat diets cut down both the expression of CD36, a protein with high affinity for fatty acids present in the apical membrane of enterocytes, and the expression of the signaling mechanisms of satiation induced by fat. This signal, which unlike other intestinal hormones has the ability to prolong the latency between meals, is generated mainly by the production of the lipid hormone oleoylethanolamide (OEA). This study further highlights a new additional disadvantage of high-fat diets and the existence of a correlation between different genes, suggesting that CD36 act as a fatty acids sensor and have regulator roles upon the intestinal mechanisms of OEA production