Multilayered Reprogramming in Response to Persistent DNA Damage in C. elegans.

DNA damage causally contributes to aging and age-related diseases. Mutations in nucleotide excision repair (NER) genes cause highly complex congenital syndromes characterized by growth retardation, cancer susceptibility, and accelerated aging in humans. Orthologous mutations in Caenorhabditis elegans lead to growth delay, genome instability, and accelerated functional decline, thus allowing investigation of the consequences of persistent DNA damage during development and aging in a simple metazoan model. Here, we conducted proteome, lipidome, and phosphoproteome analysis of NER-deficient animals in response to UV treatment to gain comprehensive insights into the full range of physiological adaptations to unrepaired DNA damage. We derive metabolic changes indicative of a tissue maintenance program and implicate an autophagy-mediated proteostatic response. We assign central roles for the insulin-, EGF-, and AMPK-like signaling pathways in orchestrating the adaptive response to DNA damage. Our results provide insights into the DNA damage responses in the organismal context.

Doppler Ultrasound Flow Evaluation of the Uterine Arteries Significantly Correlates with Tumor Size in Cervical Cancer Patients.

PURPOSE: The aim of this present study was to evaluate the sonographic correlation between Doppler flow characteristics of the uterine arteries and tumor size in patients with cervical cancer, in order to establish a new potential marker to monitor treatment response. METHODS: This was a retrospective cohort study of 25 patients who underwent a sonographic evaluation of Doppler flow characteristics of the uterine arteries before surgery or radiochemotherapy for early and locally advanced/advanced cervical cancer, respectively, was analyzed. The primary outcome was the correlation between Doppler flow characteristics of the uterine arteries and tumor size in patients with cervical cancer. RESULTS: Median age was 49 (range 26-85) years, and mean tumor size was 40.8 ± 17 mm. A significant positive correlation was found between tumor diameter and the uterine artery end-diastolic velocity (r = 0.47, p < 0.05) as well as the peak systolic velocity (r = 0.41, p < 0.05). No correlation was found between tumor size and the pulsatility index or resistance index. CONCLUSIONS: In cervical cancer, uterine artery velocity parameters are associated with tumor size. This finding could become particularly useful in the follow-up of locally advanced cervical cancer patients undergoing radiochemotherapy or in corroborating the selection of women with more possibility of a high response rate during neoadjuvant chemotherapy before surgery.

DAF-16/FOXO and EGL-27/GATA promote developmental growth in response to persistent somatic DNA damage.

Genome maintenance defects cause complex disease phenotypes characterized by developmental failure, cancer susceptibility and premature ageing. It remains poorly understood how DNA damage responses function during organismal development and maintain tissue functionality when DNA damage accumulates with ageing. Here we show that the FOXO transcription factor DAF-16 is activated in response to DNA damage during development, whereas the DNA damage responsiveness of DAF-16 declines with ageing. We find that in contrast to its established role in mediating starvation arrest, DAF-16 alleviates DNA-damage-induced developmental arrest and even in the absence of DNA repair promotes developmental growth and enhances somatic tissue functionality. We demonstrate that the GATA transcription factor EGL-27 co-regulates DAF-16 target genes in response to DNA damage and together with DAF-16 promotes developmental growth. We propose that EGL-27/GATA activity specifies DAF-16-mediated DNA damage responses to enable developmental progression and to prolong tissue functioning when DNA damage persists.

cAMP-dependent protein kinase A selects the excited state of the membrane substrate phospholamban.

Phosphorylation of membrane proteins is a central regulatory and signaling mechanism across cell compartments. However, the recognition process and phosphorylation mechanism of membrane-bound substrates by kinases are virtually unknown. cAMP-dependent protein kinase A (PKA) is a ubiquitous enzyme that phosphorylates several soluble and membrane-bound substrates. In cardiomyocytes, PKA targets phospholamban (PLN), a membrane protein that inhibits the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA). In the unphosphorylated state, PLN binds SERCA, reducing the calcium uptake and generating muscle contraction. PKA phosphorylation of PLN at S16 in the cytoplasmic helix relieves SERCA inhibition, initiating muscle relaxation. Using steady-state kinetic assays, NMR spectroscopy, and molecular modeling, we show that PKA recognizes and phosphorylates the excited, membrane-detached R-state of PLN. By promoting PLN from a ground state to an excited state, we obtained a linear relationship between rate of phosphorylation and population of the excited state of PLN. The conformational equilibrium of PLN is crucial to regulate the extent of PLN phosphorylation and SERCA inhibition.

Functional analysis in public schools: a summary of 90 functional analyses.

Several review and epidemiological studies have been conducted over recent years to inform behavior analysts of functional analysis outcomes. None to date have closely examined demographic and clinical data for functional analyses conducted exclusively in public school settings. The current paper presents a data-based summary of 90 functional analyses conducted in public school settings from 2006 through 2009 for 69 students. Specifically, we present data on gender, age, race, diagnosis, topography of target behaviors, number of conditions, duration of sessions, duration of analysis, functional outcomes, setting, and person serving the role of therapist. Results suggest that functional analyses in schools are possible, practical, and produce results that are comparable to those in past research.

Nematode sbRNAs: homologs of vertebrate Y RNAs.

Stem-bulge RNAs (sbRNAs) are a group of small, functionally yet uncharacterized noncoding RNAs first described in C. elegans, with a few homologous sequences postulated in C. briggsae. In this study, we report on a comprehensive survey of this ncRNA family in the phylum Nematoda. Employing homology search strategies based on both sequence and secondary structure models and a computational promoter screen we identified a total of 240 new sbRNA homologs. For the majority of these loci we identified both promoter regions and transcription termination signals characteristic for pol-III transcripts. Sequence and structure comparison with known RNA families revealed that sbRNAs are homologs of vertebrate Y RNAs. Most of the sbRNAs show the characteristic Ro protein binding motif, and contain a region highly similar to a functionally required motif for DNA replication previously thought to be unique to vertebrate Y RNAs. The single Y RNA that was previously described in C. elegans, however, does not show this motif, and in general bears the hallmarks of a highly derived family member.

Backbone NMR resonance assignment of the catalytic subunit of cAMP-dependent protein kinase A in complex with AMP-PNP.

The catalytic subunit of protein kinase A is involved with a number of signal transduction pathways and has been used as a benchmark to study the structural biology and biochemistry for the entire kinase family of enzymes. Here, we report the backbone assignment of the intact 41 kDa catalytic subunit bound to AMP-PNP.

Targeted gene knockout reveals a role in meiotic recombination for ZHP-3, a Zip3-related protein in Caenorhabditis elegans.

The meiotically expressed Zip3 protein is found conserved from Saccharomyces cerevisiae to humans. In baker's yeast, Zip3p has been implicated in synaptonemal complex (SC) formation, while little is known about the protein's function in multicellular organisms. We report here the successful targeted gene disruption of zhp-3 (K02B12.8), the ZIP3 homolog in the nematode Caenorhabditis elegans. Homozygous zhp-3 knockout worms show normal homologue pairing and SC formation. Also, the timing of appearance and the nuclear localization of the recombination protein Rad-51 seem normal in these animals, suggesting proper initiation of meiotic recombination by DNA double-strand breaks. However, the occurrence of univalents during diplotene indicates that C. elegans ZHP-3 protein is essential for reciprocal recombination between homologous chromosomes and thus chiasma formation. In the absence of ZHP-3, reciprocal recombination is abolished and double-strand breaks seem to be repaired via alternative pathways, leading to achiasmatic chromosomes and the occurrence of univalents during meiosis I. Green fluorescent protein-tagged C. elegans ZHP-3 forms lines between synapsed chromosomes and requires the SC for its proper localization.

Increasing variety of foods consumed by blending nonpreferred foods into preferred foods.

UNLABELLED: A treatment with differential or noncontingent reinforcement and nonremoval of the spoon increased the acceptance of one or two of 16 foods for 2 participants with severe food refusal. These differential levels of acceptance were demonstrated empirically in an ABAB design in which A was the presentation of the accepted (preferred) foods and B was the presentation of foods the participants refused (nonpreferred foods). Subsequently, we implemented a blending treatment that consisted of mixing (blending) nonpreferred foods into preferred foods in various ratios (e.g., 10% nonpreferred/90% preferred, 20% nonpreferred/80% preferred). We then presented nonpreferred foods that had been exposed to blending to determine if consumption of nonpreferred foods would increase following the blending treatment. We also conducted periodic reversals in which we presented nonpreferred foods that had not been exposed to the blending treatment. Following initial implementation of the blending treatment, consumption was high for nonpreferred foods that had been blended and low for nonpreferred foods that had not been blended. Consumption increased for all foods (i.e., foods that had been exposed to blending and foods that had not been exposed to blending) after seven or eight foods had been exposed to the blending treatment. Thus, the variety of foods consumed by the participants increased from one or two to 16. These results are discussed in terms of stimulus fading, conditioned food preferences, and escape extinction. DESCRIPTORS: conditioned food preferences, food refusal, negative reinforcement, stimulus fading

Training parents to implement pediatric feeding protocols.

Four different multicomponent training packages were evaluated to increase the treatment integrity of parents implementing pediatric feeding protocols. In Study 1 we exposed 3 parents to a training package that consisted of written protocols (baseline), verbal instructions, therapist modeling, and rehearsal training. Results suggested that the package was successful in increasing treatment integrity of the feeding protocols to high levels. Study 2 investigated three different parent-training packages comprised of components used in Study 1. Two parents were exposed to written protocols, verbal instructions, and modeling; 2 parents were exposed to written protocols, verbal instructions, and rehearsal; and 2 parents were exposed to written protocols and verbal instructions. Results of Study 2 showed that each parent-training package produced very high treatment integrity. Follow-up data in the clinic and home for 5 participants suggested that the results were durable for up to 3 months. These results demonstrate a first step in the transfer and application of research findings into routine clinical practice because we evaluated several methods for training parents to implement behavioral feeding protocols, and we demonstrated that these methods resulted in high levels of treatment integrity in a controlled clinical setting.

Translating multiple assessment techniques into an intervention selection model for classrooms.

Translating current research to school-based clinical practice highlights issues not often encountered in laboratory settings. With the assistance of a consultant, teachers conducted functional analyses, brief multielement treatment comparisons, and controlled treatment evaluations under naturalistic conditions in the classroom. Teachers also provided input on treatment selection. Treatment integrity data collected throughout the study suggested that teachers implemented analyses and treatments with high integrity. The functional analysis outcomes combined with effectiveness and acceptability data led to the selection of interventions that reduced problem behavior in the classrooms for each of 3 children.

The influence of therapist attention on self-injury during a tangible condition.

This study investigated the effects of therapist attention on the self-injurious behavior (SIB) of a 6-year-old girl with developmental disabilities. After results of a functional analysis indicated that SIB was maintained by attention and tangible reinforcement, tangible conditions with and without contingent verbal attention were compared. Results suggested that the inclusion of verbal attention in a tangible condition may confound functional analysis outcomes for behavior that is maintained by attention.