Intron and Small RNA Localization in Mammalian Neurons

RNA molecules are diverse in form and function. They include messenger RNAs (mRNAs) that are templates for proteins, splice products such as introns that can generate functional noncoding RNAs, and a slew of smaller RNAs such as transfer RNAs (tRNAs) that help decode mRNAs into proteins. RNAs can show distinct patterns of subcellular localization that play an important role in protein localization. However, RNA distribution in cells is incompletely understood, with prior studies focusing primarily on RNAs that are long (\u3e200 nucleotides), fully processed, and polyadenylated. We examined the distribution of RNAs in neurons. Neuronal compartments can be separated by long distances and play distinct roles, raising the possibility that RNA localization is especially overt and functionally meaningful in these cells. In our exploration, we physically dissected projections from cell bodies of neurons from the rat brain and sequenced total RNA. We describe two main findings. First, we identified excised introns that are enriched in neuronal projections and confirmed their localization by single- molecule fluorescence in situ hybridization. These are a previously unknown set of circular RNAs in neuronal projections: tailless lariats that possess a non- canonical C branchpoint. Second, we observed a highly abundant population of small (20-150 nucleotide) RNAs in neuronal projections, most of which are tRNAs. For both circular introns and tRNAs, we did not observe known RNA localization signals. Thus, many types of RNA, if sufficiently stable, appear free to diffuse to distant locations, their localization perhaps aided by the movement of large organelles in the confines of neuronal projections. Our survey of RNA molecules across subcellular compartments provides a foundation for investigating the function of these molecules and the mechanisms that localize them

Free circular introns with an unusual branchpoint in neuronal projections

The polarized structure of axons and dendrites in neuronal cells depends in part on RNA localization. Previous studies have looked at which polyadenylated RNAs are enriched in neuronal projections or at synapses, but less is known about the distribution of non-adenylated RNAs. By physically dissecting projections from cell bodies of primary rat hippocampal neurons and sequencing total RNA, we found an unexpected set of free circular introns with a non-canonical branchpoint enriched in neuronal projections. These introns appear to be tailless lariats that escape debranching. They lack ribosome occupancy, sequence conservation, and known localization signals, and their function, if any, is not known. Nonetheless, their enrichment in projections has important implications for our understanding of the mechanisms by which RNAs reach distal compartments of asymmetric cells

ORIGINAL ARTICLE: Status of Methicillin Resistant Staphylococcus aureus Infections and Evaluation of PVL Producing Strains in Belgaum, South India

Background: Panton Valentine Leukocidin (PVL) toxin, responsible for increased virulence and more severe infections can be found in both Methicillin-sensitive and Methicillin-resistant strains of Staphylococcus aureus (MSSA and MRSA). Aims & Objectives: To generate baseline data on the extent of MRSA infections and to estimate the frequency of PVL-positive S.aureus in Belgaum, South India. Material & Methods: 70 clinical isolates of S.aureus were obtained from various laboratories in Belgaum city. Theseisolates were identified, phenotypically characterized as MRSA/MSSA by disc diffusion method using oxacillin discs (1 µg) and genetically by multiplex PCR for mecA and fem B genes. PCR was subsequently carried out on all isolates to detect LukS-PV and LukF-PV genes, the markers for potential producers of PVL toxin. Results: 27 out of 70 isolates (38.6%) were confirmed as MRSA by PCR formecA. The prevalence of PVL gene was 85.1% and 48.8% in MRSA and MSSA respectively. The overall prevalence of PVL positive S.aureuswas 62.85%. Conclusion: Our study showed high percentage of PVL positive MRSA and MSSA, higher than the most reports worldwide. In the backdrop of bacterial strains gaining multiple drug resistance, our study warrants further epidemiological studies in hospitals and community levels in the region

Passively transferred human NMO-IgG exacerbates demyelination in mouse experimental autoimmune encephalomyelitis

BACKGROUND: Neuromyelitis optica (NMO) is a devastating inflammatory disorder of the optic nerves and spinal cord characterized by frequently recurring exacerbations of humoral inflammation. NMO is associated with the highly specific NMO-IgG biomarker, an antibody that binds the aquaporin-4 water channel. Aquaporin-4 is present on glial endfeet in the central nervous system (CNS). In humans, the NMO-IgG portends more frequent exacerbations and a worse long-term clinical outcome. METHODS: We tested the longer-term outcome of mice with EAE injected with NMO-IgG and followed them for 60 days. Clinical exams and pathology of the spinal cord and optic nerves were compared to mice that received control human IgG. RESULTS: Passively transferred human NMO-IgG leads to more severe neurology disability over two months after onset of disease. Clinical worsening is associated with an increased concentration of large demyelinating lesions primarily to subpial AQP4-rich regions of the spinal cord. CONCLUSIONS: NMO-IgG is pathogenic in the context of EAE in mice

Autorregulación y trabajo autónomo del estudiante en una actividad de aprendizaje basada en TIC

Este estudio analiza las estrategias de autorregulación en una práctica basada en las TIC con trabajo autónomo del estudiante (TAE). Participaron 53 estudiantes del primer curso de psicología de la Universidad Autónoma de Barcelona que realizaron prácticas de atención y percepción. Los objetivos, contenidos, materiales y evaluación de la práctica se diseñaron y desarrollaron en un entorno en línea. Posteriormente, se aplicaron varios cuestionarios con la finalidad de identificar las autovaloraciones que sobre la actividad autorregulatoria y el nivel de participación manifestaron los estudiantes. Los resultados muestran que los componentes de la dimensión cognitiva de la autorregulación (concepciones de aprendizaje y estrategias metacognitivas) se relacionan significativamente entre sí; sin embargo, no explican las diferencias encontradas en la valoración que los estudiantes realizan de su participación activa. Los factores del componente motivacional (orientación y autoeficacia) se encuentran, igualmente, relacionados entre sí, pero sólo la autoeficacia explica las diferencias observadas en la valoración de la participación activaThis work analyses a self-regulation strategies in autonomous task in an ICT-based learning activity. Participants were 53 psychology undergraduates at the Universidad Autónoma de Barcelona who made a practice in Attention and Perception Psychology. The aims, contents, materials and assessment of the practice were implemented in a virtual environment. We applied several questionnaires in order to identify the students' self-assessments about their self-regulatory activity and participation level. The results showed that cognitive dimension components in the self-regulation construct (conception of learning and metacognitive strategies) are significantly interrelated. However, this dimension does not account for the differences found in self-assessments students' active participation. With regard to motivational dimension factors (goal orientation and self-efficacy), a significant relation between them is observed, but only self-efficacy explains the differences found in self-assessments' active participatio

Response surface methodology for lovastatin production by Aspergillus terreus GD13 strain

A wild type Aspergillus terreus GD13 strain, chosen after extensive screening, was optimized for lovastatin production using statistical Box-Behnken design of experiments. The interactive effect of four process parameters, i.e. lactose and soybean meal, inoculum size (spore concentration) and age of the spore culture, on the production of lovastatin was evaluated employing response surface methodology (RSM). The model highlighted the positive effect of soybean meal concentration and inoculum level for achieving maximal level of lovastatin (1342 mg/l). The optimal fermentation conditions improved the lovastatin titre by 7.0-folds when compared to the titres obtained under unoptimized conditions

Screening and selection of lovastatin hyper-producing mutants of Aspergillus terreus using cyclic mutagenesis

134 fungal cultures isolated from different soil samples were screened for lovastatin production. Of these, 38 isolates produced different levels of lovastatin. An Aspergillus terreus strain GD13 , producing 190 mg/l of lovastatin was selected and subjected to a rational mutation-selection programme based on the resistance to lovastatin and fatty acid synthase (FAS) inhibitors, viz., iodoacetamide and N-ethylmaleimide. After three cycles of mutagenesis, a hyper-producing mutant (EM19) exhibiting 7.5-fold (1424 mg/l) higher levels of lovastatin when compared to wild type parent strain was obtained

Machine Learning Quantification of Amyloid Deposits in Histological Images of Ligamentum Flavum

Wild-type transthyretin amyloidosis (ATTRwt) is an underdiagnosed and potentially fatal disease. Interestingly, ATTRwt deposits have been found to deposit in the ligamentum flavum (LF) of patients with lumbar spinal stenosis before the development of systemic and cardiac amyloidosis. In order to study this phenomenon and its possible relationship with LF thickening and systemic amyloidosis, a precise method of quantifying amyloid deposits in histological slides of LF is critical. However, such a method is currently unavailable. Here, we present a machine learning quantification method with Trainable Weka Segmentation (TWS) to assess amyloid deposition in histological slides of LF. Images of ligamentum flavum specimens stained with Congo red are obtained from spinal stenosis patients undergoing laminectomies and confirmed to be positive for ATTRwt. Amyloid deposits in these specimens are classified and quantified by TWS through training the algorithm via user-directed annotations on images of LF. TWS can also be automated through exposure to a set of training images with user-directed annotations, and then applied] to a set of new images without additional annotations. Additional methods of color thresholding and manual segmentation are also used on these images for comparison to TWS. We develop the use of TWS in images of LF and demonstrate its potential for automated quantification. TWS is strongly correlated with manual segmentation in the training set of images with user-directed annotations (R = 0.98; p = 0.0033) as well as in the application set of images where TWS was automated (R = 0.94; p = 0.016). Color thresholding was weakly correlated with manual segmentation in the training set of images (R = 0.78; p = 0.12) and in the application set of images (R = 0.65; p = 0.23). TWS machine learning closely correlates with the gold-standard comparator of manual segmentation and outperforms the color thresholding method. This novel machine learning method to quantify amyloid deposition in histological slides of ligamentum flavum is a precise, objective, accessible, high throughput, and powerful tool that will hopefully pave the way towards future research and clinical applications