Splicing Dysregulation as Oncogenic Driver and Passenger Factor in Brain Tumors

Brain tumors are a heterogeneous group of neoplasms ranging from almost benign to highly aggressive phenotypes. The malignancy of these tumors mostly relies on gene expression reprogramming, which is frequently accompanied by the aberrant regulation of RNA processing mechanisms. In brain tumors, defects in alternative splicing result either from the dysregulation of expression and activity of splicing factors, or from mutations in the genes encoding splicing machinery components. Aberrant splicing regulation can generate dysfunctional proteins that lead to modification of fundamental physiological cellular processes, thus contributing to the development or progression of brain tumors. Herein, we summarize the current knowledge on splicing abnormalities in brain tumors and how these alterations contribute to the disease by sustaining proliferative signaling, escaping growth suppressors, or establishing a tumor microenvironment that fosters angiogenesis and intercellular communications. Lastly, we review recent efforts aimed at developing novel splicing-targeted cancer therapies, which employ oligonucleotide-based approaches or chemical modulators of alternative splicing that elicit an impact on brain tumor biology

c-MYC empowers transcription and productive splicing of the oncogenic splicing factor Sam68 in cancer

The splicing factor Sam68 is upregulated in many human cancers, including prostate cancer (PCa) where it promotes cell proliferation and survival. Nevertheless, in spite of its frequent upregulation in cancer, the mechanism(s) underlying its expression are largely unknown. Herein, bioinformatics analyses identified the promoter region of the Sam68 gene (KHDRBS1) and the proto-oncogenic transcription factor c-MYC as a key regulator of Sam68 expression. Upregulation of Sam68 and c-MYC correlate in PCa patients. c-MYC directly binds to and activates the Sam68 promoter. Furthermore, c-MYC affects productive splicing of the nascent Sam68 transcript by modulating the transcriptional elongation rate within the gene. Importantly, c-MYC-dependent expression of Sam68 is under the tight control of external cues, such as androgens and/or mitogens. These findings uncover an unexpected coordination of transcription and splicing of Sam68 by c-MYC, which may represent a key step in PCa tumorigenesis

Innovation and telemedicine: an applied project

Abstract.Telemedicine, defined by the World Health Organization as 'integrative part of telecommunication systems in the promotion of public health', generates different positive results: more efficiency, less costs, better patient's quality of life. Thanks to the advances in diagnostic technologies, information technologies, remote monitoring and long-distance care, telemedicine have increased the viability of home-based care, even for patients with serious conditions. At the same time we observe the increase in the elderly population suffering from pluripatologyand chronic diseases, that causes an increasing recourse to medical and hospital care; but prolonged hospital stay can determine loss of function and psychophysical disorders. Telemedicine is 'the investigation, monitoring and management of patients and education of patients and staff using systems which allow ready access to expert advice and patient information, no matter where the patient or relevant information is located'. We are empowering in different areas new biomedical and information technology as a support to clinical practice. The goal is to make a case study and to use for future analysis. In the experimental phase 4 patients were followed on hemodialysis and 4 in peritoneal dialysis, previously evaluated for clinical, social, logistical and psychological conditions. It is estimated that today there are 50.000 patients on dialysis and about 10.000 new cases a year only in Italy. Naturally there are threats to solve because users face difficulties, in terms of interaction with robots, in their usual context of life.The right methodology needs to coordinate the entire hospital, different professionals, patients, providing tools for care givers that follow patients, training technicians, aligning data collection and communication.The project reached great results in relation to the indicators: efficiency in dialysis, compliance with health protocols, impact on patient quality of life. Telemedicine and telecardiology projects are active at the 'Hospital at Home Service of Bari'.Key words: health, telemedicine, home-based care,hospital at home, robotics, ict

DNA Damage Regulates the Functions of the RNA Binding Protein Sam68 through ATM-Dependent Phosphorylation

Simple Summary Alterations of the complex network of interactions between the DNA damage response pathway and RNA metabolism have been described in several tumors, and increasing efforts are devoted to the elucidation of the molecular mechanisms involved in this network. Previous large-scale proteomic studies identified the RNA binding protein Sam68 as a putative target of the ATM kinase. Herein, we demonstrate that ATM phosphorylates Sam68 upon DNA damage induction, and this post-translational modification regulates both the signaling function of Sam68 in the initial phase of the DNA damage response and its RNA processing activity. Thus, our study uncovers anew crosstalk between ATM and Sam68, which may represent a paradigm for the functional interaction between the DDR pathway and RNA binding proteins, and a possible actionabletarget in human cancers. Cancer cells frequently exhibit dysregulation of the DNA damage response (DDR), genomic instability, and altered RNA metabolism. Recent genome-wide studies have strongly suggested an interaction between the pathways involved in the cellular response to DDR and in the regulation of RNA metabolism, but the molecular mechanism(s) involved in this crosstalk are largely unknown. Herein, we found that activation of the DDR kinase ATM promotes its interaction with Sam68, leading to phosphorylation of this multifunctional RNA binding protein (RBP) on three residues: threonine 61, serine 388 and serine 390. Moreover, we demonstrate that ATM-dependent phosphorylation of threonine 61 promotes the function of Sam68 in the DDR pathway and enhances its RNA processing activity. Importantly, ATM-mediated phosphorylation of Sam68 in prostate cancer cells modulates alternative polyadenylation of transcripts that are targets of Sam68, supporting the notion that the ATM-Sam68 axis exerts a multifaceted role in the response to DNA damage. Thus, our work validates Sam68 as an ATM kinase substrate and uncovers an unexpected bidirectional interplay between ATM and Sam68, which couples the DDR pathway to modulation of RNA metabolism in response to genotoxic stress

Splicing targeting drugs highlight intron retention as an actionable vulnerability in advanced prostate cancer

BackgroundAdvanced prostate cancer (PC) is characterized by insensitivity to androgen deprivation therapy and chemotherapy, resulting in poor outcome for most patients. Thus, advanced PC urgently needs novel therapeutic strategies. Mounting evidence points to splicing dysregulation as a hallmark of advanced PC. Moreover, pharmacologic inhibition of the splicing process is emerging as a promising option for this disease.MethodBy using a representative androgen-insensitive PC cell line (22Rv1), we have investigated the genome-wide transcriptomic effects underlying the cytotoxic effects exerted by three splicing-targeting drugs: Pladienolide B, indisulam and THZ531. Bioinformatic analyses were performed to uncover the gene structural features underlying sensitivity to transcriptional and splicing regulation by these treatments. Biological pathways altered by these treatments were annotated by gene ontology analyses and validated by functional experiments in cell models.ResultsAlthough eliciting similar cytotoxic effects on advanced PC cells, Pladienolide B, indisulam and THZ531 modulate specific transcriptional and splicing signatures. Drug sensitivity is associated with distinct gene structural features, expression levels and cis-acting sequence elements in the regulated exons and introns. Importantly, we identified PC-relevant genes (i.e. EZH2, MDM4) whose drug-induced splicing alteration exerts an impact on cell survival. Moreover, computational analyses uncovered a widespread impact of splicing-targeting drugs on intron retention, with enrichment in genes implicated in pre-mRNA 3'-end processing (i.e. CSTF3, PCF11). Coherently, advanced PC cells displayed high sensitivity to a specific inhibitor of the cleavage and polyadenylation complex, which enhances the effects of chemotherapeutic drugs that are already in use for this cancer.ConclusionsOur study uncovers intron retention as an actionable vulnerability for advanced PC, which may be exploited to improve therapeutic management of this currently incurable disease

Analysis of Secreted Proteins from Prepubertal Ovarian Tissues Exposed In Vitro to Cisplatin and LH

It is well known that secreted and exosomal proteins are associated with a broad range of physiological processes involving tissue homeostasis and differentiation. In the present paper, our purpose was to characterize the proteome of the culture medium in which the oocytes within the primordial/primary follicles underwent apoptosis induced by cisplatin (CIS) or were, for the most part, protected by LH against the drug. To this aim, prepubertal ovarian tissues were cultured under control and in the presence of CIS, LH, and CIS + LH. The culture media were harvested after 2, 12, and 24 h from chemotherapeutic drug treatment and analyzed by liquid chromatography-mass spectrometry (LC-MS). We found that apoptotic conditions generated by CIS in the cultured ovarian tissues and/or oocytes are reflected in distinct changes in the extracellular microenvironment in which they were cultured. These changes became evident mainly from 12 h onwards and were characterized by the inhibition or decreased release of a variety of compounds, such as the proteases Htra1 and Prss23, the antioxidants Prdx2 and Hbat1, the metabolic regulators Ldha and Pkm, and regulators of apoptotic pathways such as Tmsb4x. Altogether, these results confirm the biological relevance of the LH action on prepuberal ovaries and provide novel information about the proteins released by the ovarian tissues exposed to CIS and LH in the surrounding microenvironment. These data might represent a valuable resource for future studies aimed to clarify the effects and identify biomarkers of these compounds' action on the developing ovary

Body piercing and tattoos: a survey on young adults' knowledge of the risks and practices in body art

<p>Abstract</p> <p>Background</p> <p>The practice of tattooing and piercing has expanded in western society. In order to verify young adults' knowledge of the risk and practices related to body art, an investigation was conducted among freshmen of the University of Bari in the region of Apulia, Italy.</p> <p>Methods</p> <p>The study was carried out in the Academic Year 2009-2010 through an anonymous self-administered written questionnaire distributed to 1.656 freshmen enrolled in 17 Degree Courses.</p> <p>Results</p> <p>Of the 1.598 students included in the analysis, 78.3% believe it is risky to undergo piercing/tattoo practices. AIDS was indicated as a possible infection by 60.3% of freshmen, hepatitis C by 38.2%, tetanus by 34.3% and hepatitis B by 33.7% of the sample. 28.1% of freshmen were not aware that there are also non-infectious complications. 29% of the sample had at least one piercing or tattoo (this percentage does not include earlobe piercing in women). Of those with body art, the decision to undergo body art was made autonomously in 57.9% of the participants. 56.3% of freshmen undergoing body art had taken less than a month to decide. With regard to the reasons that led the sample to undergo body art, 28.4% were unable to explain it, 23.8% answered to improve their aesthetic aspect, 18.4% to distinguish themselves from others, 12.3% for fashion; 17.1% for other reasons. 25.4% of the sample declared that they had a piercing (79.8% female vs 20.2% male; ratio M/F 1:4.0). The average age for a first piercing was 15.3 years (range 10-27; SD ± 2.9). 9.6% of the sample declared that they have a tattoo (69.9% female vs 30.1% male; ratio M/F 1:2.3). The average age for a first tattoo was 17.5 years (range 10-26, SD ± 2.4).</p> <p>Conclusions</p> <p>Most of the freshmen knew about AIDS-related risks but not other potential risks. Body art is fairly common among young adults (especially women). The decision is often not shared with the family and is undertaken mostly without a specific reason or for the improvement of aesthetic aspect. Information about freshmen's knowledge, attitudes and practices could help in effective planning of health promotion strategies.</p

Antifungal susceptibility of invasive yeast isolates in Italy: the GISIA3 study in critically ill patients

<p>Abstract</p> <p>Background</p> <p>Yeasts are a common cause of invasive fungal infections in critically ill patients. Antifungal susceptibility testing results of clinically significant fungal strains are of interest to physicians, enabling them to adopt appropriate strategies for empiric and prophylactic therapies. We investigated the antifungal susceptibility of yeasts isolated over a 2-year period from hospitalised patients with invasive yeast infections.</p> <p>Methods</p> <p>638 yeasts were isolated from the blood, central venous catheters and sterile fluids of 578 patients on general and surgical intensive care units and surgical wards. Etest strips and Sensititre panels were used to test the susceptibility of the isolates to amphotericin B, anidulafungin, caspofungin, fluconazole, itraconazole, posaconazole and voriconazole in 13 laboratories centres (LC) and two co-ordinating centres (CC). The Clinical and Laboratory Standards Institute (CLSI) reference broth microdilution method was used at the CCs for comparison.</p> <p>Results</p> <p>Etest and Sensititre (LC/CC) MIC₉₀values were, respectively: amphotericin B 0.5/0.38, 1/1 mg/L; anidulafungin 2/1.5 and 1/1 mg/L; caspofungin 1/0.75 and 0.5/0.5 mg/L; fluconazole 12/8 and 16/16 mg/L; itraconazole 1/1.5, 0.5/0.5 mg/L; posaconazole 0.5 mg/L and voriconazole 0.25 mg/L for all. The overall MIC₉₀values were influenced by the reduced susceptibility of <it>Candida parapsilosis </it>isolates to echinocandins and a reduced or lack of susceptibility of <it>Candida glabrata </it>and <it>Candida krusei </it>to azoles, in particular fluconazole and itraconazole. Comparison of the LC and CC results showed good Essential Agreement (90.3% for Etest and 92.9% for Sensititre), and even higher Categorical Agreement (93.9% for Etest and 96% for Sensititre); differences were observed according to the species, method, and antifungal drug. No cross-resistance between echinocandins and triazoles was detected.</p> <p>Conclusions</p> <p>Our data confirm the different antifungal susceptibility patterns among species, and highlight the need to perform antifungal susceptibility testing of clinically relevant yeasts. With the exception of a few species (e.g. <it>C. glabrata </it>for azoles and <it>C. parapsilosis </it>for echinocandins), the findings of our study suggest that two of the most widely used commercial methods (Etest and Sensititre) provide valid and reproducible results.</p

Resource efficient low power laser cleaning of compact discs for material reuse by polycarbonate recovery

This paper deals with laser cleaning of compact discs with the aim to recover the polycarbonate layers and allow their reuse for further applications. A low power 30 W MOPFA and Q-switched pulsed Yb:YAG fibre laser is employed to ablate the metal substrate located between the polycarbonate layer and the outer serigraphy of the CDs. Compared to traditional cleaning processes, this method offers low environmental impact and greater flexibility of use. Two experimental test series are carried out to investigate the interaction mechanisms involved in the process and the effect of the process parameters on the mechanism of separation. The final aim is to identify the process conditions that could ensure a 100% cleaned surface without polymer degradation and evaluate the process time required to clean a CD

Juiz constitucional : Estado e poder no século XXI : homenagem ao Ministro Enrique Ricardo Lewandowski

- Divulgação dos SUMÁRIOS das obras recentemente incorporadas ao acervo da Biblioteca Ministro Oscar Saraiva do STJ. Em respeito à lei de Direitos Autorais, não disponibilizamos a obra na íntegra.- Localização na estante: 342 J93