BCL6 and LRF crosstalk in follicular lymphoma

B cell Lymphoma 6 (BCL6) and Leukaemia/Lymphoma related factor (LRF) are Pokproteins over-expressed in some types of Non-Hodgkin\u27s lymphoma. BCL6 is located on chromosome 3 in the breakpoint affecting 3q27 band which is the most frequent translocation in Non-Hodgkin\u27s lymphomas. This gene is a transcriptional repressor whose principal effect is to thwart the response to DNA damage by directly inhibiting both p53 and the cell cycle inhibitor p21. In lymphoma cells BCL6 prevents apoptosis induced by DNA damage. LRF, encoded by the Zbtb7a gene, also known as Pokemon, is a transcriptional repressor involved in many cellular processes as viral infection, differentiation, inflammation and oncogenesis. LRF plays an important role as proto-oncogene in Non- Hodgkin\u27s lymphomas: LRF indirectly inhibits p53, by repressing p14/ARF with consequent activation of MDM2 followed by p53 degradation. LRF is often aberrantly over-expressed in association with BCL6 in diffuse large B cell lymphoma (DLBCL) and follicular lymphoma (FL), the commonest types of Non- Hodgkin\u27s lymphomas. It has been demonstrated that microRNAs, a class of endogenous 22-25 nt single stranded RNA molecules, regulate target gene expression at the post-transcriptional level by binding with imperfect complementarity to specific regions of the 3\u27UTR of the target mRNA and act by repressing its translation. Recently miRNAs were shown to participate in the network of oncogenes and tumour suppressors, behaving either as tumour suppressors or as oncogenes, with the consequent implication of their potential use as novel anti tumourigenic drug. For these reason we investigated the role of miRNAs in the complex network connecting BCL6 and LRF in follicular lymphoma. Results In this work we focused our attention on the possible correlation between BCL6 and LRF in DOHH2 cells taking into consideration miRNAs which potentially link these two oncogenes. Using our data and data present in the literature, we reconstructed a hypothetical circuit connecting BCL6 and LRF. A negative feedback loop appears to link BCL6 to LRF, however in DLBCL and FL both genes are over expressed; these data strongly indicate that the negative feedback loop is bypassed in tumour cells to favour high proliferation rate. Different approaches were used to modulate the expression of BCL6 and LRF in Dohh2, a cell line derived from Follicular Lymphoma. A) Dohh2 cells were treated with etoposide, a chemotherapeutic agent that is able to reduce both BCL6 and LRF. Our results show that the reduction of BCL-6/LRF cells is followed by cell proliferation block and over-expression of miR-145, a tumour suppressor miRNA, under p53 control, which targets c-myc. Interestingly while c-myc was down regulated as expected, E2F1 resulted to be up regulated. B) To verify whether miR-145 could substitute for etoposide treatment miR-145 was transiently over expressed in Dohh2 cells. Although the network was influenced by miR-145 and both BCL6 and LRF were down regulated, no change in the proliferation rate of Dohh2 cells was found. Unexpectedly p53 was also markedly down regulated which could in part explain why cell proliferation was not inhibited notwithstanding BCL6/LRF decrease. C) In order to see the effect of a prolonged increase of miR-145, stably BCL6 silenced Dohh2 cells were used. In this case miR-145 was stably up regulated due to p53 increase and LRF was down regulated, yet again no effect on cell proliferation was observed. Our results clearly show that in the tumour Dohh2 cells BCL6 and LRF are always co-regulated (contrary to the hypothetical network); however their concomitant down regulation is not sufficient to determine cell proliferation block as cells appear to find a new equilibrium to counteract these anti-proliferative signals. We suggest that the down regulation of BCL6/LRF must be concomitant to the upregulation of the tumour suppressor p53 and the proto-oncogene E2F1 to influence the cell cycle and that miRNAs (such as miR-145 and miR-20) are part of these regulatory mechanisms

Immortalization of MEF is characterized by the deregulation of specific miRNAs with potential tumor suppressor activity

The life span (Hayflick limit) of primary mouse embryo fibroblasts (MEF) in culture is variable but it is still unclear if the escape of the Hayflick limit is also variable. To address this point MEF were expanded every fifteen days (6T15) instead of every three days (6T3) until they became immortal. With this protocol MEF lifespan was extended and immortalization accordingly delayed. By testing a panel of genes (p19ARF, p16, p21) and miRNAs (miR-20a, miR-21, miR-28, miR-290) related to primary MEF senescence, a switch of p21 from up to down regulation, the down regulation of specific miRNAs as well as a massive shift from diploidy to hyperdiploidy were observed in coincidence with the resumption of cell proliferation. Collectively, these data indicate that the inactivation of genes and miRNAs, important in controlling cell proliferation, might be determinant for the escape from the Hayflick limit. In support of this hypothesis was the finding that some of the down regulated miRNAs transfected in immortalized MEF inhibited cell proliferation thus displaying a tumor suppressor-like activity

Immortalized mouse embryo fibroblasts are resistant to miR-290-induced senescence regardless of p53 status

Immortalized mouse embryo fibroblasts are resistant to miR-290-induced senescence regardless of p53 status. Physiol Genomics 43: 1153-1159, 2011. First published August 16, 2011; doi:10.1152/physiolgenomics.00064.2011.-The prosenescence role of miR-290 and nocodazole has been documented in primary mouse embryo fibroblasts (MEF), while it is not clear whether immortal murine fibroblasts are still responsive to these senescence inducing stimuli. To establish this point, immortal murine fibroblasts with functional (NIH3T3) or nonfunctional p53 (I-MEF) and low levels of miR-290 were tested for their capability to undergo senescence after exposure to either nocodazole or miR-290. Our results clearly indicate that nocodazole induces senescence only in NIH3T3 cells with a functional p53 but not in I-MEF lacking a functional p53. miR-290 overexpression is unable to address any of the tested immortalized clones toward senescence, regardless of the p53 status, suggesting that the prosenescence role of miR-290 is specific for primary but not for immortal murine fibroblasts. Moreover our findings suggest that the mere downregulation of a potential tumor suppressor miRNA in a given cell type does not necessarily imply that it behaves as a tumor suppressor

O uso do conhecimento como técnica evolutiva voltada à melhoria dos processos e de tomada de decisão

In the current scenario of organizations Process Management aims to perform process in an organized, prioritizing actions through techniques and methods that are linked to the improvement of the organization in the market segment it operates. As a determinant for achieving goals successfully, organizational memory is all important in the management of processes, allowing all areas meet in a systemic way, crediting their information to the various organizational sectors and thus using knowledge to direct an action with a focus planning to achieve strategic organizational goal. The organizational memory can be registered through the development of a knowledge base. The work in question aims to provide a better understanding of the importance of the knowledge base in an organization, to perform appropriate actions, planning, simulating and reaching a decision through meaningful data. Soon after the development of a bibliographic research, a bibliometric study on the proposed topic was accomplished with the main events of the Brazilian scientific areas of production engineering, analyzing how the topic has been addressed by authors in the areas of Process Management and Base Knowledge. The main results so far observed the need for the implementation of knowledge-based models in systems that seek to improve the execution of processes and thus reduce failures and decision-making processes more appropriate.No atual cenário das organizações, a Gestão por Processos tem como foco realizar processos de forma organizada, priorizando ações por meio de técnicas e métodos que estejam vinculados à melhoria da organização no segmento de mercado onde atua. Como fator determinante para atingir metas com sucesso, a memória organizacional é de total importância no gerenciamento dos processos, possibilitando atender a todas as áreas de forma sistêmica, creditando suas informações aos diversos setores organizacionais e, assim, utilizando o conhecimento para dirigir uma ação com foco no planejamento para alcançar um objetivo estratégico organizacional. A memória organizacional pode ser registrada por meio do desenvolvimento de uma base de conhecimento. O trabalho em questão tem como objetivo permitir uma melhor compreensão sobre a importância da base de conhecimento em uma organização, para realizar ações apropriadas, planejando, simulando e alcançando uma decisão através de dados significativos. Logo após o desenvolvimento de uma pesquisa bibliográfica, um estudo bibliométrico sobre o tema proposto foi realizado junto aos principais eventos científicos brasileiros das áreas de engenharia de produção, analisando-se como o assunto tem sido abordado por autores das áreas da Gestão por Processos e Base de Conhecimento. Nos principais resultados encontrados até o momento, observou-se a necessidade da implantação de modelos baseados em conhecimento, em sistemas que buscam a melhoria na execução dos processos e consequente diminuição de falhas e processos de tomada de decisão mais adequados

The Proto-Oncogene LRF Is under Post-Transcriptional Control of MiR-20a: Implications for Senescence

MicroRNAs (miRNAs) are short 20–22 nucleotide RNA molecules that act as negative regulators of gene expression via translational repression: they have been shown to play a role in development, proliferation, stress response, and apoptosis. The transcriptional regulator LRF (Leukemia/lymphoma Related Factor) has been shown to prevent p19ARF transcription and consequently to inhibit senescence in mouse embryonic fibroblasts (MEF). Here we report, for the first time, that LRF is post-transcriptionally regulated by miR-20a. Using a gene reporter assay, direct interaction of miR-20a with the LRF 3′UTR is demonstrated. To validate the interaction miR-20a/3′UTR LRF miR-20a was over-expressed, either by transient transfection or retroviral infection, in wild type mouse embryo fibroblasts and in LRF-null MEF derived from LRF knock-out mice. We observed LRF decrease, p19ARF increase, inhibition of cell proliferation and induction of senescence. The comparison of miR-20a activity in wt and LRF-null MEF indicates that LRF is the main mediator of the miR-20a-induced senescence and that other targets are cooperating. As LRF down-regulation/p19ARF induction is always accompanied by E2F1 down-regulation and increase of p16, we propose that all these events act in synergy to accomplish miR-20a-induced senescence in MEF. Senescence has been recently revaluated as a tumor suppressor mechanism, alternative to apoptosis; from this point of view the discovery of new physiological “senescence inducer” appears to be promising as this molecule could be used as anticancer drug

Lavaggio delle mani: confronto tra comportamenti dei Professionisti e degli Studenti in un grande Ospedale universitario

RIASSUNTO L'obiettivo principale dello studio è valutare le conoscenze, le opinioni e il rispetto delle procedure di operatori sanitari (medici, infermieri, studenti di medici e di infermieristica) sull'igiene delle mani (HH). Numerose ricerche indicano una minor accuratezza nel rispetto delle procedure di igiene della mani comparata agli infermieri mentre poche ricerche indagano l'atteggiamento durante gli studi. Un questionario è stato somministrato ad un campione di 756 partecipanti: 252 medici, 268 infermieri, 117 studenti infermieristica, 119 studenti di medicina in un grande Ospedale universitario (Roma), per determinare la conoscenza, la conformití  e le procedure rispetto all'igiene delle mani. Per l'analisi delle risposte dei questionari è stato utilizzato SPSS 17.0. La conoscenza e l'aderenza alle procedure dei futuri medici è inferiore a quella dei futuri infermieri. Sia nell'HBS (opinioni sull'igiene della mani) che nell'HHPI (procedure nell'igiene delle mani), gli studenti infermieri hanno punteggi più elevati rispetto agli infermieri, ai medici e agli studenti di medicina. Le domande sull'uso di detergenti a base di alcool sono state quelle in cui vi è il più basso numero di risposte corrette, in tutte le professioni. I medici hanno una minore aderenza alla pratica dell'igiene delle mani, rispetto gli infermieri. Future ricerche dovrebbe chiarire quali siano le differenze nella costruzione dello studio sulla pratica dell'igiene delle mani, quali sono le barriere che impediscono l' aderenza dei sanitari alla stessa e le migliori metodiche per trasmettere l'abitudine all'igiene della mani. I professionisti dovrebbero inoltre essere maggiormente coinvolti nella lotta contro le infezioni nosocomiali (ICA). Parole chiave: igiene delle mani, infezioni associate all'assistenza ICA, student di medicina, studenti infermieri, studenti delle professioni sanitarie, compliance nell'igiene delle mani, procedure per l'igiene delle mani ABSTRACT The main objective of the study is assessing knowledge, opinions and compliance with the procedures of health professionals (physician, nurses, medical and nursing students) about Hand Hygiene (HH). There is a number of research which indicates that physicians respect less than nurses Hand Hygiene, there are a smaller number which investigates the differences in the attitude of the aforementioned subjects during their studies. A questionnaire was administered to a sample of 756 participants (252 doctors, 268 nurses, 117 nursing students and 119 students of medicine) at a large University Hospital in Rome, to determine their knowledge, compliance and procedures with Hand Hygiene. Knowledge of medical students is lower than that of nursing students, as well as they have lower values in adherence to practice. In both HH Beliefs Scale (HBS) and HH Practicies Inventory (HHPI) questionnaires, nursing students have higher scores than nurses, doctors and medical students. The questions on the use of alcohol-based cleaners have been those where there was the lowest number of correct responses, across all professions. Physicians compared to nurses have a lower adherence to Hand Hygiene. Future research should clarify what the differences are in the construction of the study on the practice of washing hands, what are the barriers to health professionals and best methods for teaching habits, namely the effectiveness of hand washing. Professionals should be more involved in the fight against Healthcare-associated infections. Key words: hand hygiene, health care associated infection, medical students. Nursing students, health care professionals, hand hygiene compliance, hand hygiene procedure

CONSEQUÊNCIAS DAS INTERVENÇÕES ANTRÓPICAS NA NASCENTE DA MURITIBA EM LENÇÓIS, BAHIA

Este trabalho teve como objetivo estabelecer uma relação entre a situação da nascente da Muritiba, localizada na cidade de Lençóis, Bahia e as intervenções humanas por ela sofrida ao longo dos anos. A pesquisa buscou demonstrar como as intervenções se estabeleceram e foram determinantes para a ocorrência das alterações ecológicas. Após uma visita de campo e observações do local, foram levantadas diversas hipóteses, sob olhares de profissionais de diversas áreas, sobre as possíveis causas da nascente, antes permanente, ter se tornado temporária. Historicamente, a facilidade de acesso e a qualidade da água, fez com que a população buscasse a nascente, principalmente nos períodos de seca. Entretanto, a captação de águas subterrâneas no entorno da nascente, inviabilizou a disponibilidade de água, gerando questões socioambientais relevantes. Dessa forma, o estudo apontou a necessidade de atividades do Poder Público local e de Educação Ambiental com vista a proteção ambiental da área

Clinical relevance of postzygotic mosaicism in Cornelia de Lange syndrome and purifying selection of NIPBL variants in blood

Postzygotic mosaicism (PZM) in NIPBL is a strong source of causality for Cornelia de Lange syndrome (CdLS) that can have major clinical implications. Here, we further delineate the role of somatic mosaicism in CdLS by describing a series of 11 unreported patients with mosaic disease-causing variants in NIPBL and performing a retrospective cohort study from a Spanish CdLS diagnostic center. By reviewing the literature and combining our findings with previously published data, we demonstrate a negative selection against somatic deleterious NIPBL variants in blood. Furthermore, the analysis of all reported cases indicates an unusual high prevalence of mosaicism in CdLS, occurring in 13.1% of patients with a positive molecular diagnosis. It is worth noting that most of the affected individuals with mosaicism have a clinical phenotype at least as severe as those with constitutive pathogenic variants. However, the type of genetic change does not vary between germline and somatic events and, even in the presence of mosaicism, missense substitutions are located preferentially within the HEAT repeat domain of NIPBL. In conclusion, the high prevalence of mosaicism in CdLS as well as the disparity in tissue distribution provide a novel orientation for the clinical management and genetic counselling of families.Spanish Ministry of Health-ISCIII Fondo de Investigación Sanitaria (FIS) [Ref. PI19/01860, to F.J.R. and J.P.]; Diputación General de Aragón-FEDER: European Social Fund [Grupo de Referencia B32_17R / B32_20R, to J.P.]. A.L-P is supported by a “Juan de la Cierva-Incorporación” postdoctoral grant from MICIU (Spanish Ministry of Science and Universities

Protocolos de regulação ambulatorial : dermatologia

Telemedicin

Lack of SARS-CoV-2 RNA environmental contamination in a tertiary referral hospital for infectious diseases in Northern Italy

none140noNAnoneColaneri M.; Seminari E.; Piralla A.; Zuccaro V.; Di Filippo A.; Baldanti F.; Bruno R.; Mondelli M.U.; Brunetti E.; Di Matteo A.; Maiocchi L.; Pagnucco L.; Mariani B.; Ludovisi S.; Lissandrin R.; Parisi A.; Sacchi P.; Patruno S.F.A.; Michelone G.; Gulminetti R.; Zanaboni D.; Novati S.; Maserati R.; Orsolini P.; Vecchia M.; Sciarra M.; Asperges E.; Sambo M.; Biscarini S.; Lupi M.; Roda S.; Chiara Pieri T.; Gallazzi I.; Sachs M.; Valsecchi P.; Perlini S.; Alfano C.; Bonzano M.; Briganti F.; Crescenzi G.; Giulia Falchi A.; Guarnone R.; Guglielmana B.; Maggi E.; Martino I.; Pettenazza P.; Pioli di Marco S.; Quaglia F.; Sabena A.; Salinaro F.; Speciale F.; Zunino I.; De Lorenzo M.; Secco G.; Dimitry L.; Cappa G.; Maisak I.; Chiodi B.; Sciarrini M.; Barcella B.; Resta F.; Moroni L.; Vezzoni G.; Scattaglia L.; Boscolo E.; Zattera C.; Michele Fidel T.; Vincenzo C.; Vignaroli D.; Bazzini M.; Iotti G.; Mojoli F.; Belliato M.; Perotti L.; Mongodi S.; Tavazzi G.; Marseglia G.; Licari A.; Brambilla I.; Daniela B.; Antonella B.; Patrizia C.; Giulia C.; Giuditta C.; Marta C.; Rossana D.; Milena F.; Bianca M.; Roberta M.; Enza M.; Stefania P.; Maurizio P.; Elena P.; Antonio P.; Francesca R.; Antonella S.; Maurizio Z.; Guy A.; Laura B.; Ermanna C.; Giuliana C.; Luca D.; Gabriella F.; Gabriella G.; Alessia G.; Viviana L.; Claudia L.; Valentina M.; Simona P.; Marta P.; Alice B.; Giacomo C.; Irene C.; Alfonso C.; Di Martino R.; Di Napoli A.; Alessandro F.; Guglielmo F.; Loretta F.; Federica G.; Alessandra M.; Federica N.; Giacomo R.; Beatrice R.; Maria S.I.; Monica T.; Nepita Edoardo V.; Calvi M.; Tizzoni M.; Nicora C.; Triarico A.; Petronella V.; Marena C.; Muzzi A.; Lago P.; Comandatore F.; Bissignandi G.; Gaiarsa S.; Rettani M.; Bandi C.Colaneri, M.; Seminari, E.; Piralla, A.; Zuccaro, V.; Di Filippo, A.; Baldanti, F.; Bruno, R.; Mondelli, M. U.; Brunetti, E.; Di Matteo, A.; Maiocchi, L.; Pagnucco, L.; Mariani, B.; Ludovisi, S.; Lissandrin, R.; Parisi, A.; Sacchi, P.; Patruno, S. F. A.; Michelone, G.; Gulminetti, R.; Zanaboni, D.; Novati, S.; Maserati, R.; Orsolini, P.; Vecchia, M.; Sciarra, M.; Asperges, E.; Sambo, M.; Biscarini, S.; Lupi, M.; Roda, S.; Chiara Pieri, T.; Gallazzi, I.; Sachs, M.; Valsecchi, P.; Perlini, S.; Alfano, C.; Bonzano, M.; Briganti, F.; Crescenzi, G.; Giulia Falchi, A.; Guarnone, R.; Guglielmana, B.; Maggi, E.; Martino, I.; Pettenazza, P.; Pioli di Marco, S.; Quaglia, F.; Sabena, A.; Salinaro, F.; Speciale, F.; Zunino, I.; De Lorenzo, M.; Secco, G.; Dimitry, L.; Cappa, G.; Maisak, I.; Chiodi, B.; Sciarrini, M.; Barcella, B.; Resta, F.; Moroni, L.; Vezzoni, G.; Scattaglia, L.; Boscolo, E.; Zattera, C.; Michele Fidel, T.; Vincenzo, C.; Vignaroli, D.; Bazzini, M.; Iotti, G.; Mojoli, F.; Belliato, M.; Perotti, L.; Mongodi, S.; Tavazzi, G.; Marseglia, G.; Licari, A.; Brambilla, I.; Daniela, B.; Antonella, B.; Patrizia, C.; Giulia, C.; Giuditta, C.; Marta, C.; D'Alterio, Rossana; Milena, F.; Bianca, M.; Roberta, M.; Enza, M.; Stefania, P.; Maurizio, P.; Elena, P.; Antonio, P.; Francesca, R.; Antonella, S.; Maurizio, Z.; Guy, A.; Laura, B.; Ermanna, C.; Giuliana, C.; Luca, D.; Gabriella, F.; Gabriella, G.; Alessia, G.; Viviana, L.; Meisina, Claudia; Valentina, M.; Simona, P.; Marta, P.; Alice, B.; Giacomo, C.; Irene, C.; Alfonso, C.; Di Martino, R.; Di Napoli, A.; Alessandro, F.; Guglielmo, F.; Loretta, F.; Federica, G.; Albertini, Alessandra; Federica, N.; Giacomo, R.; Beatrice, R.; Maria, S. I.; Monica, T.; Nepita Edoardo, V.; Calvi, M.; Tizzoni, M.; Nicora, C.; Triarico, A.; Petronella, V.; Marena, C.; Muzzi, A.; Lago, P.; Comandatore, F.; Bissignandi, G.; Gaiarsa, S.; Rettani, M.; Bandi, C