79 research outputs found
A Case for Adult Two-Way Bilingual Immersion
No full textThe present study investigates 2-way bilingual immersion (TWBI) as a potentially viable pedagogical model for adult language learners. A review of the literature on TWBI at the K-6 level is provided, followed by an examination of key issues in adult second and foreign language education. Implications for potential adult TWBI programs are discussed along with recommendations for further investigation. Finally, the author presents an exploratory study of a nonformal, communitybased adult TWBI program in Los Angeles known as I HABLO U. The results of this study suggest that while adult TWBI shares many of the learner and administrative challenges documented in K-6 TWBI programs, adult learners in TWBI programs contend with a unique set of problems and also enjoy a number of advantages that K-6 learners may not experience. The author concludes that scholars must widen the focus of current research and evaluative efforts of TWBI to consider adult learners
High-throughput analysis reveals novel maternal germline RNAs crucial for primordial germ cell preservation and proper migration
No full textDuring oogenesis, hundreds of maternal RNAs are selectively localized to the animal or vegetal pole, including determinants of somatic and germline fates. Although microarray analysis has identified localized determinants, it is not comprehensive and is limited to known transcripts. Here, we utilized high-throughput RNA sequencing analysis to comprehensively interrogate animal and vegetal pole RNAs in the fully grown Xenopus laevis oocyte. We identified 411 (198 annotated) and 27 (15 annotated) enriched mRNAs at the vegetal and animal pole, respectively. Ninety were novel mRNAs over 4-fold enriched at the vegetal pole and six were over 10-fold enriched at the animal pole. Unlike mRNAs, microRNAs were not asymmetrically distributed. Whole-mount in situ hybridization confirmed that all 17 selected mRNAs were localized. Biological function and network analysis of vegetally enriched transcripts identified protein-modifying enzymes, receptors, ligands, RNA-binding proteins, transcription factors and co-factors with five defining hubs linking 47 genes in a network. Initial functional studies of maternal vegetally localized mRNAs show that sox7 plays a novel and important role in primordial germ cell (PGC) development and that ephrinB1 (efnb1) is required for proper PGC migration. We propose potential pathways operating at the vegetal pole that highlight where future investigations might be most fruitful.Fil: Owens, Dawn A.. University of Miami; Estados UnidosFil: Butler, Amanda M.. University of Miami; Estados UnidosFil: Agüero, Tristán Horacio. University of Miami; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; ArgentinaFil: Newman, Karen M.. University of Miami; Estados UnidosFil: Van Booven, Derek. University of Miami; Estados UnidosFil: King, Mary Lou. University of Miami; Estados Unido
The Congo as topos of dystopic transgression in fin-de-siècle literature
No full textIn this essay, I compare the representation of the Congo as a topos of dystopic transgression in Conrad’s Heart of Darkness (1902), and in a lesser-known novel entitled Tropenwee (Tropical agony) by the Dutch author Henri van Booven, published in 1904. The idea of the Congo as a locus of degeneration will be read, not so much as a Conradian theme, but rather, as an idea that had gained wide currency throughout Europe during the fin-de-siècle period. Particular attention will be paid to some of the narrative techniques that shape this idea and the ideological assumptions it conveys. Moreover, I hope to show that degeneration as reflected by the writings under investigation is at once a colonial and anti-colonial theme, and therefore its significance requires moving beyond singular and clear-cut ideological labels. 
Abstract 6053: Increased nitric oxide augments the action of CSF-1R inhibition against tumor associated macrophages in castration resistant prostate cancer
No full textAbstract Introduction and Objectives: Prostate cancer (PCa) is the most common non-skin cancer among American men, and growing evidence suggests that targeting the tumor microenvironment (TME) could be essential in combating the progression of metastasis and resistance development in cancer. In this context, the colony-stimulating factor 1 (CSF1)/colony-stimulating factor 1 receptor (CSF1R) axis has gained the most attention, and various approaches targeting either the ligands or the receptor are currently in clinical development. However, as a class, CSF1R inhibitors have proved mostly disappointing in early clinical trials when used as monotherapy. Researchers believe their true potential can be tapped by combining them with other anticancer drugs. Sensitization to CSF-1R blockage therapy is tumor microenvironment (TME) driven. In one of our studies, we demonstrated that increased nitric oxide (NO) reduces tumor burden in murine models for CRPC by targeting TME . Therefore, in the present study, we evaluated the hypothesis that Increased Nitric oxide augments the action of CSF-1R inhibition against tumor associated macrophages in castration resistant prostate cancer. Methods: The castrated SCID mice were treated with CSF1R inhibitor (GS2580) at the dosage of 40mg/kg/day IP or/and GSNO at the dosage of 10mg/kg/day IP. After 4 weeks mice were humanely sacrificed. Tumor RNA and proteins to analyze the markers that are important for prostate cancer progression using qPCR, western blot and cytokine antibody array. RNA library was generated, and RNA sequencing was performed using RNA isolated from tumors. Molecular analyses were performed using standard procedures. GraphPad Prism (GraphPad Software) was used for statistical analysis. All data were presented as the means ± SEM. Results: Mice which recieved CSF1R inhibition showed significant reduction in tumor burden (p<0.05), however, in over 50% of the mice, the expression of markers like AR, pERK, p-GSK, and VEGF was found to be increased. Next, to study if increased Nitric oxide levels are able to augment the action of CSF1Ri against CRPC, we studied the effects of GSNO monotherapy, CSF1Ri monotherapy and GSNO+CSF1Ri combination on overall tumor burden. Results revealed that the most significant reduction in tumor burden were in mice that recieved the combination of GSNO-CSF1Ri, compared to GSNO or CSF1Ri monotherpies. Furthermore, RNA sequencing analysis demonstrated that the combination therapy is capable of targeting (suppressing) tumor immunology (signature of interferon-alpha, gama, and Myc signaling were significantly reduced). This was further supported by cytokine antibody array and immunostaning which showed that several cytokines like CXCL5, FGF4, IGFBP-3, MCP-4, IL-6, TNFalpha, expression of CRRPC markers- AR, ARV7, PSA, TMRPSS2, p-GSK, p-ERK, p90RSK, and markers of anti-inflamatorry macrophages (F4/80, CD206 etc) were suppressed in tumor proteins from mice which received combination therapy. Conclusions: Our findings suggest that CSF1R inhibition induced changes against CRPC are augmented in the presence of increased NO levels therefore demonstrating the therapeutic potential of increased NO levels against CRPC. Citation Format: Yash Soni, Manish Kuchakulla, Rehana Qureshi, Van Booven Derek J, Joshua M. Hare, Ranjith Ramasamy, Himanshu Arora. Increased nitric oxide augments the action of CSF-1R inhibition against tumor associated macrophages in castration resistant prostate cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6053
Dysregulation of the histone demethylase KDM6B in alcohol dependence is associated with epigenetic regulation of inflammatory signaling pathways
No full textEpigenetic enzymes oversee long-term changes in gene expression by integrating genetic and environmental cues. While there are hundreds of enzymes that control histone and DNA modifications, their potential roles in substance abuse and alcohol dependence remain underexplored. A few recent studies have suggested that epigenetic processes could underlie transcriptomic and behavioral hallmarks of alcohol addiction. In the present study, we sought to identify epigenetic enzymes in the brain that are dysregulated during protracted abstinence as a consequence of chronic and intermittent alcohol exposure. Through quantitative mRNA expression analysis of over 100 epigenetic enzymes, we identified 11 that are significantly altered in alcohol-dependent rats compared with controls. Follow-up studies of one of these enzymes, the histone demethylase KDM6B, showed that this enzyme exhibits region-specific dysregulation in the prefrontal cortex and nucleus accumbens of alcohol-dependent rats. KDM6B was also upregulated in the human alcoholic brain. Upregulation of KDM6B protein in alcohol-dependent rats was accompanied by a decrease of trimethylation levels at histone H3, lysine 27 (H3K27me3), consistent with the known demethylase specificity of KDM6B. Subsequent epigenetic (chromatin immunoprecipitation [ChIP]–sequencing) analysis showed that alcohol-induced changes in H3K27me3 were significantly enriched at genes in the IL-6 signaling pathway, consistent with the well-characterized role of KDM6B in modulation of inflammatory responses. Knockdown of KDM6B in cultured microglial cells diminished IL-6 induction in response to an inflammatory stimulus. Our findings implicate a novel KDM6B-mediated epigenetic signaling pathway integrated with inflammatory signaling pathways that are known to underlie the development of alcohol addiction
mitoTALEN Eliminates Mutant mtDNA Genomes in Neurons
No full textMutations in the mitochondrial DNA (mtDNA) commonly cause severe encephalopathies. Because most of these mtDNA alterations are heteroplasmic, we used a mitochondrial-targeted TALEN (mitoTALEN) to specifically eliminate the mutant mtDNA in the CNS of a mouse model harboring a heteroplasmic mutation in the mitochondrial tRNA alanine gene (m.5024C>T). Delivery to neurons was achieved by using AAV-PHP.eB and neuronal expression was obtained by using a neuronal-specific synapsin promoter. We found that most CNS regions were effectively transduced and showed a significant reduction in mutant mtDNA. This reduction was accompanied by an increase in mitochondrial tRNA alanine level, which is drastically reduced by the mutation. These results showed, for the first time, that mitochondrial-targeted gene editing can be effective in reducing CNS mutant mtDNA in vivo, paving the way for clinical trials in patients with mitochondrial encephalopathies
Author Correction: Defective HNF4alpha-dependent gene expression as a driver of hepatocellular failure in alcoholic hepatitis
No full textAuthor Correction to: "Defective HNF4alpha-dependent gene expression as a driver of hepatocellular failure in alcoholic hepatitis
Additional file 4: Table S4. of Epigenomic and metabolic responses of hypothalamic POMC neurons to gestational nicotine exposure in adult offspring
No full textCo-expressed lncRNAs and protein-coding genes in cis. Co-expression of lncRNAs (columns A and B) with their nearest (adjacent or overlapping) protein-coding gene (column D) was determined by the Pearson’s correlation coefficient r (column F). An r value > 0.602 or 1 CPM (columns K and L) were considered for analyses. LncRNA expression was in most cases less than that of the coding gene (column M). Coding genes function in various biological processes (column N). (XLSX 63 kb
Whole Genome Sequencing and a New Bioinformatics Platform Allow for Rapid Gene Identification in D. melanogaster EMS Screens
No full textCo-author Jeannette Osterloh is a doctoral student in the Neuroscience program in the Morningside Graduate School of Biomedical Sciences (GSBS) at UMass Medical School. Forward genetic screens in Drosophila melanogaster using ethyl methanesulfonate (EMS) mutagenesis are a powerful approach for identifying genes that modulate specific biological processes in an in vivo setting. The mapping of genes that contain randomly-induced point mutations has become more efficient in Drosophila thanks to the maturation and availability of many types of genetic tools. However, classic approaches to gene mapping are relatively slow and ultimately require extensive Sanger sequencing of candidate chromosomal loci. With the advent of new high-throughput sequencing techniques, it is increasingly efficient to directly re-sequence the whole genome of model organisms. This approach, in combination with traditional chromosomal mapping, has the potential to greatly simplify and accelerate mutation identification in mutants generated in EMS screens. Here we show that next-generation sequencing (NGS) is an accurate and efficient tool for high-throughput sequencing and mutation discovery in Drosophila melanogaster. As a test case, mutant strains of Drosophila that exhibited long-term survival of severed peripheral axons were identified in a forward EMS mutagenesis. All mutants were recessive and fell into a single lethal complementation group, which suggested that a single gene was responsible for the protective axon degenerative phenotype. Whole genome sequencing of these genomes identified the underlying gene ect4. To improve the process of genome wide mutation identification, we developed Genomes Management Application (GEM.app, https://genomics.med.miami.edu), a graphical online user interface to a custom query framework. Using a custom GEM.app query, we were able to identify that each mutant carried a unique non-sense mutation in the gene ect4 (dSarm), which was recently shown by Osterloh et al. to be essential for the activation of axonal degeneration. Our results demonstrate the current advantages and limitations of NGS in Drosophila and we introduce GEM.app as a simple yet powerful genomics analysis tool for the Drosophila community. At a current cost of < $1,000 per genome, NGS should thus become a standard gene discovery tool in EMS induced genetic forward screens.Neuroscienc
Correcting a pathogenic mitochondrial DNA mutation by base editing in mice
No full textPrimary mitochondrial disorders are most often caused by deleterious mutations in the mitochondrial DNA (mtDNA). Here, we used a mitochondrial DddA-derived cytosine base editor (DdCBE) to introduce a compensatory edit in a mouse model that carries the pathological mutation in the mitochondrial transfer RNA (tRNA) alanine (mt-tRNAAla) gene. Because the original m.5024C -> T mutation (G -> A in the mt-tRNAAla) destabilizes the mt-tRNAAla aminoacyl stem, we designed a compensatory m.5081G -> A edit (C -> T in the mt-tRNAAla) that could restore the secondary structure of the tRNAAla aminoacyl stem. For this, the DdCBE gene construct was initially tested in an m.5024C -> T mutant cell line. The reduced mt-tRNAAla amounts in these cells were increased after editing up to 78% of the mtDNA. Then, DdCBE was packaged in recombinant adeno-associated virus 9 (AAV9) and intravenously administered by retro-orbital injections into mice. Expression of the transduced DdCBE was observed in the heart and skeletal muscle. Total mt-tRNAAla amounts were restored in heart and muscle by the m.5081G -> A edit in a dose-dependent manner. Lactate amounts, which were increased in the heart, were also decreased in treated mice. However, the highest dose tested of AAV9-DdCBE also induced severe adverse effects in vivo because of the extensive mtDNA off-target editing that it generated. These results show that although DdCBE is a promising gene therapy tool for mitochondrial disorders, the doses of the therapeutic constructs must be carefully monitored to avoid deleterious off-target editing
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