Arrhythmia Management in Pediatric Patients with Ventricular Assist Devices

Ventricular assist device therapy has emerged as an important approach in the management of advanced heart failure. Atrial and ventricular arrhythmias are commonly encountered in patients with heart failure. Patients requiring ventricular assist devices are at an increased risk of arrhythmia, which may cause symptoms and significant complications. There is recent focus on the prevalence and impact of atrial and ventricular arrhythmias in patients with durable ventricular assist devices. Ventricular arrhythmias in particular have been associated with significant symptoms and worse clinical outcomes. The goal of this chapter is to outline approaches to arrhythmia management in pediatric patients with ventricular assist devices

Drosophila SPF45: A Bifunctional Protein with Roles in Both Splicing and DNA Repair

The sequence of the SPF45 protein is significantly conserved, yet functional studies have identified it as a splicing factor in animal cells and as a DNA-repair protein in plants. Using a combined genetic and biochemical approach to investigate this apparent functional discrepancy, we unify and validate both of these studies by demonstrating that the Drosophila melanogaster protein is bifunctional, with independent functions in DNA repair and splicing. We find that SPF45 associates with the U2 snRNP and that mutations that remove the C-terminal end of the protein disrupt this interaction. Although animals carrying this mutation are viable, they are nevertheless compromised in their ability to regulate Sex-lethal splicing, demonstrating that Sex-lethal is an important physiological target of SPF45. Furthermore, these mutant animals exhibit phenotypes diagnostic of difficulties in recovering from exogenously induced DNA damage. The conclusion that SPF45 functions in the DNA-repair pathway is strengthened by finding both genetic and physical interactions between SPF45 and RAD201, a previously uncharacterized member of the RecA/Rad51 protein family. Together with our finding that the fly SPF45 protein increases the survival rate of mutagen-treated bacteria lacking the RecG helicase, these studies provide the tantalizing suggestion that SPF45 has an ancient and evolutionarily conserved role in DNA repair

Discutindo a educação ambiental no cotidiano escolar: desenvolvimento de projetos na escola formação inicial e continuada de professores

A presente pesquisa buscou discutir como a Educação Ambiental (EA) vem sendo trabalhada, no Ensino Fundamental e como os docentes desta escola compreendem e vem inserindo a EA no cotidiano escolar., em uma escola estadual do município de Tangará da Serra/MT, Brasil. Para tanto, realizou-se entrevistas com os professores que fazem parte de um projeto interdisciplinar de EA na escola pesquisada. Verificou-se que o projeto da escola não vem conseguindo alcançar os objetivos propostos por: desconhecimento do mesmo, pelos professores; formação deficiente dos professores, não entendimento da EA como processo de ensino-aprendizagem, falta de recursos didáticos, planejamento inadequado das atividades. A partir dessa constatação, procurou-se debater a impossibilidade de tratar do tema fora do trabalho interdisciplinar, bem como, e principalmente, a importância de um estudo mais aprofundado de EA, vinculando teoria e prática, tanto na formação docente, como em projetos escolares, a fim de fugir do tradicional vínculo “EA e ecologia, lixo e horta”.Facultad de Humanidades y Ciencias de la Educació

Genetic and Physical Interactions between SPF45 and RAD201, a Member of the RecA/Rad51 Protein Family

<div><p>(A) <i>spf45^J23</i> does not complement the MMS mutant phenotype of <i>rad201</i>. Relative survival to adulthood of the double heterozygous mutant animals was assessed by comparing the number of experimental animals with the number of sibling controls.</p><p>(B) <i>rad201</i> splicing in wild-type and <i>spf45^J23</i> mutants. Genomic organization of <i>rad201</i> (CG2412). The position of the C-to-T transition leading to a proline-to-leucine change at position 95 in the <i>rad201¹</i> open reading frame is indicated. The <i>rad201</i> gene encodes a single transcript. RT–PCR analysis using primers that span the first and second introns demonstrates that splicing is unaffected in <i>spf45^J23</i> mutants.</p><p>(C) RAD201 interacts with SPF45 in whole-cell extracts in an RNA-independent manner. The ability of RAD201 to associate with SPF45 in whole-cell extracts was tested by GST pull-down assays followed by Western blotting. GST::RAD201, GST::U2A′, or GST alone, bound to glutathione sepharose beads, were incubated with extracts made from wild-type or <i>spf45^J23</i> embryos, followed by Western blotting using an antibody directed against SPF45 or SNF. The lane marked 5% input is a control in which the amount of extract corresponds to ∼5% of the material applied to the glutathione affinity beads. The RNase sensitivity of these interactions was tested by pretreating the extracts with a combination of RNase A and RNase T1.</p></div

SPF45 Function Is Required to Repair DNA Damage

<div><p>(A) Ectopic expression of SPF45 improves the survival rate of <i>E. coli recG</i> mutants after MMS exposure. The survival was expressed as the average-fold increase in survival over the control <i>E. coli recG</i> mutants. Error bars are standard deviations calculated from five independent experiments. An unpaired <i>t</i> test was performed to assess whether there is a statistically significant difference between survival rates of an <i>E. coli recG</i> strain expressing a D. melanogaster protein (either SPF45^J23 or SPF45⁺) and the survival rate of the same <i>E. coli recG</i> strain expressing an empty GST vector<i>.</i> Asterisks denote differences significant at the 95% level (<i>p</i> < 0.04).</p><p>(B) <i>spf45</i> animals have an MMS-sensitive mutant phenotype. Survival was assessed by comparing the number of mutant animals that survived to adulthood with the number of control siblings (<i>spf45^J23/CyO</i> or <i>Df(2Lh)D1/CyO</i>) recovered from the same cross.</p><p>(C) Damage after <i>P-element</i> excision is not repaired efficiently in an <i>spf45</i> mutant background. Eye colors of female progeny that inherited the repaired <i>P{w^a}</i> were scored. The total number of females scored is indicated below the relevant genotype. A chi square test was performed to assess whether there is a significant difference between the frequency of repair events. Asterisks denote differences significant at the 99.9% level (<i>p</i> < 0.0005).</p></div

D. melanogaster SPF45 Protein Is Conserved

<p>Alignment of SPF45 proteins from D. melanogaster and humans and the DRT111 protein from A. thaliana. Identical residues are shaded. The residues that comprise the G-patch domain are shaded in red, and the residues that comprise the RRM-like motif are shaded in blue. An arrow indicates the position of the <i>P-element</i> insertion in <i>spf45^J23</i>.</p

<i>Sxl</i> Splicing Is Compromised in a <i>spf45</i> Loss-of-Function Background

<div><p>(A) Synergistic genetic interactions between <i>Sxl</i> and <i>spf45^J23</i> leads to female lethality. In these assays females of the indicated genotype were mated to either <i>Sxl^f1/Y</i> males or <i>Sxl⁺/Y</i> control males and the resulting progeny scored. On the assumption that an equal number of male and female progeny should be generated from each cross, the percent female viability was calculated by comparing the number of females recovered with the number of males recovered.</p><p>(B) Diagram of the <i>Sxl</i> reporter construct that mirrors native <i>Sxl</i> splicing in all tissues tested. The arrows below the construct indicate the position of the PCR pairs used for RT–PCR.</p><p>(C) Synergistic lethal interactions between <i>Sxl</i> and <i>spf45^J23</i> are correlated with <i>Sxl</i> splicing defects. Splicing was assayed by an RT–PCR-based assay using RNA isolated from a pool of embryos in which only the female embryos carry the reporter construct (lanes 3 and 4). In lane 4, this pool of embryos was collected from <i>spf45^J23</i> homozygous females crossed to males carrying an X chromosome that carries both <i>Sxl^f1</i> and a copy of the <i>Sxl</i> reporter construct. Controls include splicing of the reporter construct in adult males (lane 1), splicing of the reporter construct in adult females (lane 2), and <i>Sxl⁺</i> embryos collected from <i>spf45^J23</i> homozygous mothers (lane 3).</p></div