Grades 3-5 Genres in Literature

This is an English language arts lesson for grades three through five on studying genres in literature. Through this lesson students will gain an understanding of the literary elements of each genre, improve comprehension by interpreting, analyzing, synthesizing, and evaluating written text in order to categorize. Students will also be able to identify characteristics of different genres, select genres that interest them, and be able to compare and contrast different works of literature with each other. This lesson spans over ten days and offers a variety of projects using a tic-tack-toe chart on three tiered levels where students can choose their favorite activity in each subject area to create a line based on their interests

Book Highlight: Winning the Customer

vol. 3, no.

Methods for Improving Efficiency of Planned Missing Data Designs

Any survey specifically constructed so that at least some variables are unobserved on a subset of participants is a planned missing data design, where missing data represent an intentional feature of the study. Use of planned missing data designs can potentially reduce costs, improve data quality, and reduce unplanned missing data, and advancements in missing data methodology and multiple imputation software make planned missing data designs more attractive than before. Two commonly used planned missing data designs are two-phase sampling and split questionnaire design. Two-phase sampling is used to improve the efficiency of an estimate for a single outcome that is costly to measure, while the split questionnaire design is primarily used to reduce survey length. First, we propose new methods for selecting our second phase sample in two-phase surveys to reduce the variance of our estimate. When our outcome variable is continuous, we can use the data collected in Phase I for selecting our Phase II sample in order to increase the precision of the estimates. For other instances, we propose an adaptive sampling method to select Phase II samples in order to improve estimation of the quantity of interest. Next, we examine the performance of several design allocations for implementing a split questionnaire survey in a longitudinal study. While many papers examined the administration of split questionnaire designs in cross-sectional studies, research in applying these methods to longitudinal studies has been limited. For our project, we focus on the commonly used 3-form design and propose several methods to administer the 3-form split questionnaire in a longitudinal study. Using simulations and data from the Health and Retirement Study, we compare the performance of each proposed design under several correlation structures. Finally, we propose a method for improving variable allocation in split questionnaire designs. We establish a criterion that allows us to determine which variable allocations minimize the loss of information due to missing data. We use the Kullback-Leibler divergence between the posterior distribution of the parameters with missing data and the posterior distribution without missing data to locate optimal designs. After establishing a criterion for comparing designs, we propose a search algorithm to find optimal variable allocations, as it would be difficult to enumerate all possible designs as the number of variables grows.PHDBiostatisticsUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttps://deepblue.lib.umich.edu/bitstream/2027.42/144155/1/pimbri_1.pd

Histone Marks-Dependent Effect on Alternative Splicing: New Perspectives for Targeted Splicing Modulation in Cancer?

Alternative splicing (AS) is a tightly regulated mechanism that generates the complex human proteome from a small number of genes. Cis-regulatory RNA motifs in exons and introns control AS, recruiting positive and negative trans-acting splicing regulators. At a higher level, chromatin affects splicing events. Growing evidence indicates that the popular histone code hypothesis can be extended to RNA-level processes, such as AS. In addition to nucleosome positioning, which can generate transcriptional barriers to shape the final splicing outcome, histone post-translational modifications can contribute to the detailed regulation of single exon inclusion/exclusion. A histone-based system can identify alternatively spliced chromatin stretches, affecting RNAPII elongation locally or recruiting splicing components via adaptor complexes. In tumor cells, several mechanisms trigger misregulated AS events and produce cancer-associated transcripts. On a genome-wide level, aberrant AS can be the consequence of dysfunctional epigenetic splicing code, including altered enrichment in histone post-translational modifications. This review describes the main findings related to the effect of histone modifications and variants on splicing outcome and how a dysfunctional epigenetic splicing code triggers aberrant AS in cancer. In addition, it highlights recent advances in programmable DNA-targeting technologies and their possible application for AS targeted epigenetic modulation

Dynamic recruitment of transcription factors and epigenetic changes on the ER stress response gene promoters

Response to stresses that alter the function of the endoplasmic reticulum is an important cellular function, which relies on the activation of specific genes. Several transcription factors (TFs) are known to affect this pathway. Using RT–PCR and ChIP assays, we studied the recruitment of promoter-specific TFs, general TFs and epigenetic marks in activated promoters. H3-K4 di- and tri-methylation and H3-K79 di-methylation are present before induction. H3 acetylation is generally high before induction, and H4 acetylation shows a promoter-specific increase. Interestingly, there is a depletion of histone H3 under maximal induction, explaining an apparent decrease of H3-K4 tri-methylation and H3-K79 di-methylation. Pol II is found enriched on some promoters under basal conditions, unlike TBP and p300, which are recruited selectively. Most genes are bound by XBP-1 after induction, some before induction, presumably by the inactive isoform. ATF6 and CHOP associate to largely different set of genes. C/EBPβ is selective and binding to the CHOP promoter precedes that of XBP-1, ATF6 and CHOP. Finally, one of the ER-stress inducible genes analyzed, HRD1, is not bound by any of these factors. Among the constitutive TFs, NF-Y, but not Sp1, is found on all genes before induction. Intriguingly, siRNA interference of the NF-YB subunit indicates transcriptional impairment of some, but not all genes. These data highlight a previously unappreciated complexity of TFs binding and epigenetic changes, pointing to different TFs-specific pathways within this broad response

Transcription Factors in Cancer: When Alternative Splicing Determines Opposite Cell Fates

Alternative splicing (AS) is a finely regulated mechanism for transcriptome and proteome diversification in eukaryotic cells. Correct balance between AS isoforms takes part in molecular mechanisms that properly define spatiotemporal and tissue specific transcriptional programs in physiological conditions. However, several diseases are associated to or even caused by AS alterations. In particular, multiple AS changes occur in cancer cells and sustain the oncogenic transcriptional program. Transcription factors (TFs) represent a key class of proteins that control gene expression by direct binding to DNA regulatory elements. AS events can generate cancer-associated TF isoforms with altered activity, leading to sustained proliferative signaling, differentiation block and apoptosis resistance, all well-known hallmarks of cancer. In this review, we focus on how AS can produce TFs isoforms with opposite transcriptional activities or antagonistic functions that severely impact on cancer biology. This summary points the attention to the relevance of the analysis of TFs splice variants in cancer, which can allow patients stratification despite the presence of interindividual genetic heterogeneity. Recurrent TFs variants that give advantage to specific cancer types not only open the opportunity to use AS transcripts as clinical biomarkers but also guide the development of new anti-cancer strategies in personalized medicine

Attenuation of choroidal tickness in patients with Alzheimer disease: evidence from an Italian prospective study

INTRODUCTION: To compare the 12-month choroidal thickness (CT) change between Alzheimer disease (AD) patients and normal subjects. METHODS: In this prospective, observational study, 39 patients with a diagnosis of mild to moderate AD and 39 age-matched control subjects were included. All the subjects underwent neuropsychological (Mini Mental State Examination, Alzheimer disease Assessment Scale-Cognitive Subscale, and the Clinical Dementia Rating Scale) and ophthalmological evaluation, including spectral domain optical coherence tomography, at baseline and after 12 months. CT was measured manually using the caliper tool of the optical coherence tomography device. RESULTS: After 12 months, AD patients had a greater reduction of CT than controls (P≤0.05, adjusted for baseline CT, age, sex, axial length, and smoking). DISCUSSION: CT in patients with AD showed a rate of thinning greater than what could be expected during the natural course of aging

In situ immunofluorescent staining of autophagy in muscle stem cells

Increasing evidence points to autophagy as a crucial regulatory process to preserve tissue homeostasis. It is known that autophagy is involved in skeletal muscle development and regeneration, and the autophagic process has been described in several muscular pathologies and agerelated muscle disorders. A recently described block of the autophagic process that correlates with the functional exhaustion of satellite cells during muscle repair supports the notion that active autophagy is coupled with productive muscle regeneration. These data uncover the crucial role of autophagy in satellite cell activation during muscle regeneration in both normal and pathological conditions, such as muscular dystrophies. Here, we provide a protocol to monitor the autophagic process in the adult Muscle Stem Cell (MuSC) compartment during muscle regenerative conditions. This protocol describes the setup methodology to perform in situ immunofluorescence imaging of LC3, an autophagy marker, and MyoD, a myogenic lineage marker, in muscle tissue sections from control and injured mice. The methodology reported allows for monitoring the autophagic process in one specific cell compartment, the MuSC compartment, which plays a central role in orchestrating muscle regeneration