BEAMing and Droplet Digital PCR Analysis of Mutant IDH1 mRNA in Glioma Patient Serum and Cerebrospinal Fluid Extracellular Vesicles

Development of biofluid-based molecular diagnostic tests for cancer is an important step towards tumor characterization and real-time monitoring in a minimally invasive fashion. Extracellular vesicles (EVs) are released from tumor cells into body fluids and can provide a powerful platform for tumor biomarkers because they carry tumor proteins and nucleic acids. Detecting rare point mutations in the background of wild-type sequences in biofluids such as blood and cerebrospinal fluid (CSF) remains a major challenge. Techniques such as BEAMing (beads, emulsion, amplification, magnetics) PCR and droplet digital PCR (ddPCR) are substantially more sensitive than many other assays for mutant sequence detection. Here, we describe a novel approach that combines biofluid EV RNA and BEAMing RT-PCR (EV-BEAMing), as well droplet digital PCR to interrogate mutations from glioma tumors. EVs from CSF of patients with glioma were shown to contain mutant IDH1 transcripts, and we were able to reliably detect and quantify mutant and wild-type IDH1 RNA transcripts in CSF of patients with gliomas. EV-BEAMing and EV-ddPCR represent a valuable new strategy for cancer diagnostics, which can be applied to a variety of biofluids and neoplasms

Pharmacologic Targeting of Bacterial -Glucuronidase Alleviates Nonsteroidal Anti-Inflammatory Drug-Induced Enteropathy in Mice

Small intestinal mucosal injury is a frequent adverse effect caused by nonsteroidal anti-inflammatory drugs (NSAIDs). The underlying mechanisms are not completely understood, but topical (luminal) effects have been implicated. Many carboxylic acid-containing NSAIDs, including diclofenac (DCF), are metabolized to acyl glucuronides (AGs), and/or ether glucuronides after ring hydroxylation, and exported into the biliary tree. In the gut, these conjugates are cleaved by bacterial β-glucuronidase, releasing the potentially harmful aglycone. We first confirmed that DCF-AG was an excellent substrate for purified Escherichia coli β-d-glucuronidase. Using a previously characterized novel bacteria-specific β-glucuronidase inhibitor (Inhibitor-1), we then found that the enzymatic hydrolysis of DCF-AG in vitro was inhibited concentration dependently (IC50 ∼164 nM). We next hypothesized that pharmacologic inhibition of bacterial β-glucuronidase would reduce exposure of enterocytes to the aglycone and, as a result, alleviate enteropathy. C57BL/6J mice were administered an ulcerogenic dose of DCF (60 mg/kg i.p.) with or without oral pretreatment with Inhibitor-1 (10 μg per mouse, b.i.d.). Whereas DCF alone caused the formation of numerous large ulcers in the distal parts of the small intestine and increased (2-fold) the intestinal permeability to fluorescein isothiocyanate-dextran, Inhibitor-1 cotreatment significantly alleviated mucosal injury and reduced all parameters of enteropathy. Pharmacokinetic profiling of DCF plasma levels in mice revealed that Inhibitor-1 coadministration did not significantly alter the Cmax, half-life, or area under the plasma concentration versus time curve of DCF. Thus, highly selective pharmacologic targeting of luminal bacterial β-d-glucuronidase by a novel class of small-molecule inhibitors protects against DCF-induced enteropathy without altering systemic drug exposure

Arqueología cultural de los videojuegos. Entre discursos nostálgicos, experiencia del gamer e innovación tecnológica

In this paper, it is presented a videogames history as an innovation beyond a form of entertainment, offering reasons why it is important to know and investigate its history in relation to their social and cultural contexts. Marking the importance that users have when creating video games through their experience, questioning masculinity constructed in the first era of videogames and no or less participation of the female gender in the early decades. The social and cultural context in which those games were born is basic to understanding the spread and gained popularity in the ‘80s and especially in the ‘90s. The objective of this study is to identify communication strategies and information about the videogame before the arrival of the Internet and how information was shared in the Spanish context. In the first part we introduce the theoretical and methodological framework in which this research is based through the concept of cultural archeology. In the second part we present stories built by users to analyze the experiences of the game and how to share it, using the concepts of playformance and playworld. To finish questioning the identity of the gamer as a male subject, young white and upper middle class.En este trabajo, se presentara una historia de los videojuegos como una innovación más allá de una forma de entretenimiento, ofreciendo razones acerca de por qué es importante conocer e investigar su historia en relación a sus contextos sociales y culturales. Haciendo énfasis en la importancia que los usuarios tienen a la hora de crear videojuegos a través de su experiencia, cuestionando la masculinidad construida en los primeros videojuegos y la ausencia, o menor participación, del género femenino en las primeras décadas. El contexto social y cultural en que nacieron aquellos videojuegos es básico para entender la difusión y la popularidad que obtuvieron en los años ‘80 y sobre todo, en los ‘90. El objetivo de este trabajo es identificar las estrategias de comunicación y de información respecto a los videojuegos antes de la llegada de Internet, en especial la forma y el cómo se compartía esta información en el contexto español. En la primera parte introducimos el marco teórico y metodológico en que se basa esta investigación a través del concepto de arqueología cultural. En la segunda parte presentamos los relatos construidos por los usuarios para analizar las experiencias del juego y la forma de compartirlo, utilizando los conceptos de playformance y playworld. Para finalizar cuestionando la identidad del gamer como sujeto masculino, blanco y joven de clase media-alta

Atomic Energy Commission Report NYO-9077

Nuclear processes are random in character, and, accordingly, are amenable to statistical treatment. In 1905, shortly after the discover of natural radioactivity, E. Von Schweldler showed that the analytical description of the decay distribution of radioactive substances followed from probability considerations regardless of the mechanism involved in the process of atomic disintegration

Visual outcomes in children with neurofibromatosis type 1-associated optic pathway glioma following chemotherapy: a multicenter retrospective analysis

Optic pathway gliomas (OPGs) occur in 15%-20% of children with neurofibromatosis type 1 (NF1); up to half become symptomatic. There is little information regarding ophthalmologic outcomes after chemotherapy. A retrospective multicenter study was undertaken to evaluate visual outcomes following chemotherapy for NF1-associated OPG, to identify risks for visual loss, and to ascertain indications for treatment. Subjects included children undergoing initial treatment for OPGs with chemotherapy between January 1997 and December 2007. Of 115 subjects, visual acuity (VA) decline and tumor progression were the primary reasons to initiate treatment, although there were significant differences in the pattern of indications cited among the institutions. Eighty-eight subjects and 168 eyes were evaluable for VA outcome. At completion of chemotherapy, VA improved (32% of subjects), remained stable (40%), or declined (28%). Tumor location was the most consistent prognostic factor for poor VA outcome. There was poor correlation between radiographic and VA outcomes. Although visual outcomes for NF1-associated OPG are not optimal, approximately one-third of children regain some vision with treatment. Since radiographic outcomes do not predict visual outcomes, their use as the primary measure of treatment success is in question. The lack of consensus regarding the indications for treatment underlines the need for better standardization of care. Future clinical trials for OPG require standardized visual assessment methods and clear definitions of visual outcomes.</p