Elements in the Canine Distemper Virus M 3′ UTR Contribute to Control of Replication Efficiency and Virulence

Canine distemper virus (CDV) is a negative-sense, single-stranded RNA virus within the genus Morbillivirus and the family Paramyxoviridae. The Morbillivirus genome is composed of six transcriptional units that are separated by untranslated regions (UTRs), which are relatively uniform in length, with the exception of the UTR between the matrix (M) and fusion (F) genes. This UTR is at least three times longer and in the case of CDV also highly variable. Exchange of the M-F region between different CDV strains did not affect virulence or disease phenotype, demonstrating that this region is functionally interchangeable. Viruses carrying the deletions in the M 3′ UTR replicated more efficiently, which correlated with a reduction of virulence, suggesting that overall length as well as specific sequence motifs distributed throughout the region contribute to virulence

Effects of antiplatelet therapy on stroke risk by brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases: subgroup analyses of the RESTART randomised, open-label trial

Background Findings from the RESTART trial suggest that starting antiplatelet therapy might reduce the risk of recurrent symptomatic intracerebral haemorrhage compared with avoiding antiplatelet therapy. Brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases (such as cerebral microbleeds) are associated with greater risks of recurrent intracerebral haemorrhage. We did subgroup analyses of the RESTART trial to explore whether these brain imaging features modify the effects of antiplatelet therapy

Identifying profiles of multisystem physiological activity across early childhood: Examining developmental shifts and associations with stress and internalizing problems

Physiological regulation is an important predictor of health across the lifespan. Regulation occurs across multiple collaborative systems, yet few empirical studies explore multisystem activity and how this collaborative regulation develops early in life. The current study used latent profile analysis to evaluate multisystem regulation in the autonomic nervous system and hypothalamic pituitary adrenal (HPA) axis in 150 racially/ethnically diverse, low-income children at 18- and 36-months. At both timepoints, profiles of generally moderate activity (Moderate Arousal) and heightened baseline activity (Anticipatory Arousal) emerged. A profile of typically adaptive patterns across all systems (Active Copers) emerged at 18-months and a profile of heightened HPA Axis activity (HPA-axis Responders) emerged at 36-months. Persistent membership in the Anticipatory Arousal profile across time was associated with exposure to greater maternal stress at 18-months and child internalizing problems at 36-months. These findings highlight early multisystem profile development and suggest associations with stress and later behavior problems

Developmental consequences of early life stress on risk for psychopathology: Longitudinal associations with children's multisystem physiological regulation and executive functioning

The etiology of psychopathology is multifaceted and warrants consideration of factors at multiple levels and across developmental time. Although experiences of adversity in early life have been associated with increased risk of developing psychopathology, pathways toward maladaptation or resilience are complex and depend upon a variety of factors, including individuals' physiological regulation and cognitive functioning. Therefore, in a longitudinal cohort of 113 mother-child dyads, we explored associations from early adverse experiences to physiological co-regulation across multiple systems and subsequent variations in executive functioning. Latent profile analysis derived multisystem profiles based on children's heart rate, respiratory sinus arrhythmia, pre-ejection period, and cortisol measured during periods of rest and reactivity throughout a developmentally challenging protocol. Three distinct profiles of multisystem regulation emerged: heightened multisystem baseline activity (Anticipatory Arousal/ANS Responder), typically adaptive patterns across all systems (Active Copers/Mobilizers) and heightened HPA axis activity (HPA Axis Responders). Path models revealed that children exposed to adversity before 18-months were more likely to evidence an Anticipatory Arousal/ANS Responders response at 36-months, and children in this profile had lower executive functioning scores than the Active Copers/Mobilizers. In sum, these findings provide important information about potential physiological associations linking early adversity to variations in children's task-based executive functioning

Cumulative comorbid conditions influence mortality risk after staged palliation for hypoplastic left heart syndrome and variants

Objective: Prematurity, low birth weight, genetic syndromes, extracardiac conditions, and secondary cardiac lesions are considered high-risk conditions associated with mortality after stage 1 palliation. We report the impact of these conditions on outcomes from a prospective multicenter improvement collaborative. Methods: The National Pediatric Cardiology Quality Improvement Collaborative Phase II registry was queried. Comorbid conditions were categorized and quantified to determine the cumulative burden of high-risk diagnoses on survival to the first birthday. Logistic regression was applied to evaluate factors associated with mortality. Results: Of the 1421 participants, 40% (575) had at least 1 high-risk condition. The aggregate high-risk group had lower survival to the first birthday compared with standard risk (76.2% vs 88.1%, P \u3c .001). Presence of a single high-risk diagnosis was not associated with reduced survival to the first birthday (odds ratio, 0.71; confidence interval, 0.49-1.02, P = .066). Incremental increases in high-risk diagnoses were associated with reduced survival to first birthday (odds ratio, 0.23; confidence interval, 0.15-0.36, P \u3c .001) for 2 and 0.17 (confidence interval, 0.10-0.30, P \u3c .001) for 3 to 5 high-risk diagnoses. Additional analysis that included prestage 1 palliation characteristics and stage 1 palliation perioperative variables identified multiple high-risk diagnoses, poststage 1 palliation extracorporeal membrane oxygenation support (odds ratio, 0.14; confidence interval, 0.10-0.22, P \u3c .001), and cardiac reoperation (odds ratio, 0.66; confidence interval, 0.45-0.98, P = .037) to be associated with reduced survival odds to the first birthday. Conclusions: The presence of 1 high-risk diagnostic category was not associated with decreased survival at 1 year. Cumulative diagnoses across multiple high-risk diagnostic categories were associated with decreased odds of survival. Further patient accrual is needed to evaluate the impact of specific comorbid conditions within the broader high-risk categories

PRICKLE1 Interaction with SYNAPSIN I Reveals a Role in Autism Spectrum Disorders

<div><p>The frequent comorbidity of Autism Spectrum Disorders (ASDs) with epilepsy suggests a shared underlying genetic susceptibility; several genes, when mutated, can contribute to both disorders. Recently, <i>PRICKLE1</i> missense mutations were found to segregate with ASD. However, the mechanism by which mutations in this gene might contribute to ASD is unknown. To elucidate the role of PRICKLE1 in ASDs, we carried out studies in <i>Prickle1^+/−</i> mice and <i>Drosophila</i>, yeast, and neuronal cell lines. We show that mice with <i>Prickle1</i> mutations exhibit ASD-like behaviors. To find proteins that interact with PRICKLE1 in the central nervous system, we performed a yeast two-hybrid screen with a human brain cDNA library and isolated a peptide with homology to SYNAPSIN I (SYN1), a protein involved in synaptogenesis, synaptic vesicle formation, and regulation of neurotransmitter release. Endogenous Prickle1 and Syn1 co-localize in neurons and physically interact via the <i>SYN1</i> region mutated in ASD and epilepsy. Finally, a mutation in <i>PRICKLE1</i> disrupts its ability to increase the size of dense-core vesicles in PC12 cells. Taken together, these findings suggest <i>PRICKLE1</i> mutations contribute to ASD by disrupting the interaction with SYN1 and regulation of synaptic vesicles.</p></div

USIPP antibody recognizes a brain-specific, 74 kDa protein.

<p>(<b>a</b>) Anti-USIPP immunoblot shows an unknown brain-specific 74 kDa in a wild-type mouse. Lysates prepared from wild-type mouse brain, kidney or liver were resolved by SDS-PAGE and subjected to anti-USIPP Western blot analysis. (<b>b</b>) Confocal images show USIPP antibody immunostaining endogenous protein in dentate gyrus region of a wild-type mouse hippocampus. Sections of WT mouse hippocampi were incubated in anti-PSD-95 and rabbit pre-immune serum or immune USIPP serum followed by red AlexaFluor568 goat anti-mouse (PSD-95) and green AlexaFluor488 goat anti-rabbit (USIPP) secondary antibodies. Confocal images were captured with a Zeiss 710 microscope. The size markers correspond to 100 µm.</p

PRICKLE1 Yeast two-hybrid screen isolates Unknown Short Interacting Prickle1 Peptide (USIPP).

<p>Using Human PRICKLE1 (aa 1 to 827) N-LexA-PRICKLE1-C cloned into pB27 vector as bait, a yeast two-hybrid assay was used to screen random-primed adult and fetal cDNA libraries constructed into the pP6 vector. 53 million clones (adult brain library) and 99 million clones (fetal brain library) were screened using a mating approach and nutritional selection in media lacking tryptophan, leucine, and histidine. Applying a high Predicted Biological Score (PBS), nine final sequences were identified out of a total of 507 positive clones selected. Multiple sequence alignment of the peptides with Kalign® highlights an unknown 42-residue consensus peptide sequence (USIPP).</p

Endogenous SYN co-localizes with PRICKLE in the mouse brain and <i>Drosophila</i> neuromuscular junction, and coimmunoprecipitates with PRICKLE1 in mouse brain.

<p>(<b>a</b>) Co-localization of Prickle1 with Synapsin I in mouse hippocampal neurons in primary culture. Merged fluorescence image shows endogenous expression of PRICKLE1 (green) and SYNAPSIN I (red). Hippocampal neurons were fixed 10 days post-culture and immunostained with PRICKLE1 and SYNAPSIN I primary antibodies followed by AlexaFluor568 (Synapsin1) and AlexaFluor488 (Prickle1) secondary antibodies (<b>b</b>) EGFP-Pk and Synapsin co-localize at the neuromuscular junction (NMJ) in <i>Drosophila</i> third instar larvae. (I–III) 40X or (I′–III′) 100X immunohistochemistry confocal images of EGFP-Pk (I and I′) and Synapsin (II and II′) visualized at synaptic boutons of larval NMJs, along with the relevant merged images (III and III′). Note that there is a substantial co-localization of the EGFP-Pk and Synapsin signals. Scale bars = 5 µm (40X) and 5 µm (100X). Anti-GFP = rabbit anti-GFP Anti-Synapsin = mouse anti-Synapsin. (<b>c</b>) Lysates prepared from wild-type mouse brain was incubated with A/G agarose beads and USIPP antibodies, pre-immune serum or no serum overnight. Immunoprecipitates eluted in Laemmli buffer, resolved by SDS-PAGE and subjected to Western blot analyses with anti-PRICKLE1. (<b>d</b>) Lysates prepared from wild-type mouse brain was incubated with A/G agarose beads and Prickle1 antibodies, pre-immune serum or no serum overnight. Immunoprecipitates eluted in Laemmli buffer, resolved by SDS-PAGE and subjected to Western blot analyses with anti-Synapsin I.</p