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Defects in blood development frequently occur among syndromic congenital anomalies. Thrombocytopenia-Absent Radius (TAR) Syndrome is a rare congenital condition with reduced platelets (hypomegakaryocytic thrombocytopenia) and forelimb anomalies, concurrent with more variable heart and kidney defects. TAR syndrome associates with hypomorphic gene function forRBM8A/Y14that encodes a component of the exon junction complex involved in mRNA splicing, transport, and nonsense-mediated decay. How perturbing a general mRNA-processing factor causes the selective TAR Syndrome phenotypes remains unknown. Here, we connect zebrafishrbm8aperturbation to early hematopoietic defects via attenuated non-canonical Wnt/Planar Cell Polarity (PCP) signaling that controls developmental cell arrangements. In hypomorphicrbm8azebrafish, we observe a significant reduction ofcd41-positive thrombocytes.rbm8a-mutant zebrafish embryos accumulate mRNAs with individual retained introns, a hallmark of defective nonsense-mediated decay; affected mRNAs include transcripts for non-canonical Wnt/PCP pathway components. We establish thatrbm8a-mutant embryos show convergent extension defects and that reducedrbm8afunction interacts with perturbations in non-canonical Wnt/PCP pathway genes wnt5b,wnt11f2,fzd7a, andvangl2. Using live-imaging, we found reducedrbm8afunction impairs the architecture of the lateral plate mesoderm (LPM) that forms hematopoietic, cardiovascular, kidney, and forelimb skeleton progenitors as affected in TAR Syndrome. Both mutants forrbm8aand for the PCP genevangl2feature impaired expression of early hematopoietic/endothelial genes includingrunx1and the megakaryocyte regulatorgfi1aa. Together, our data propose aberrant LPM patterning and hematopoietic defects as possible consequence of attenuated non-canonical Wnt/PCP signaling upon reducedrbm8afunction. These results link TAR Syndrome to a potential LPM origin and developmental mechanism.

Due to their optical clarity and rapid development, zebrafish embryos are an excellent system for examining cell behaviors and developmental processes. However, because of the complexity and redundancy of embryonic signals, it can be challenging to discern the complete role of any single signal during early embryogenesis. By explanting the animal region of the zebrafish blastoderm, relatively naive clusters of embryonic cells are generated that can be easily cultured and manipulated ex vivo. By introducing a gene of interest by RNA injection before explantation, one can assess the effect of this molecule on gene expression, cell behaviors, and other developmental processes in relative isolation. Furthermore, cells from embryos of different genotypes or conditions can be combined in a single chimeric explant to examine cell/tissue interactions and tissue-specific gene functions. This article provides instructions for generating zebrafish blastoderm explants and demonstrates that a single signaling molecule - a Nodal ligand - is sufficient to induce germ layer formation and extension morphogenesis in otherwise naive embryonic tissues. Due to their ability to recapitulate embryonic cell behaviors, morphogen gradients, and gene expression patterns in a simplified ex vivo system, these explants are anticipated to be of great utility to many zebrafish researchers.

Background Whether the plasma proteome can predict the course of heart failure (HF) and has incremental value to established predictors is uncertain. Methods Patients meeting Framingham HF criteria with history of reduced ejection fraction (n=1017) were prospectively enrolled in a registry and donated fasting blood samples. Plasma underwent analysis on the SOMAscan proteomic discovery platform, quantifying 4789 proteins using standard assay and quality controls. Patients were randomly divided into derivation (n=681) and validation (n=336) cohorts. We derived a proteomic risk score (PRS) in the derivation cohort using Lasso-penalized Cox regression and then tested it in the validation cohort. Both models were adjusted for an establish HF clinical risk score (MAGGIC) and NTproBNP. We assessed risk stratification improvement in the validation cohort by comparing models with and without PRS using the model C statistic, continuous net reclassification index (NRI), integrated discrimination index (IDI), and the median improvement in risk score (MIRS). Results Overall 47.5% of patients were African American, 35.2% were female, mean ejection fraction was 34.8%, and average age was 67.9 years. After median follow-up of 3.6 years, there were 296 deaths (194 in derivation and 102 in validation). Optimized modeling defined a 32 protein PRS (hazard ratio [HR] 2.33, p Methods Variables Estimate 95% CI P Validation Testing MAGGIC 1.06 1.027, 1.092 2.84E-04 NTproBNP 1.84 1.430, 2.359 1.88E-06 PRS 1.19 1.001, 1.408 4.87E-02 Risk Stratification Assessment C-statistic improvement 0.012 −0.076, 0.101 8.30E-01 IDI 0.034 0.007, 0.095 Conclusion A plasma multi-protein predictive score can improve risk stratification in HF patients on top of a validated clinical score and NTproBNP. Additional investigation is warranted to define mechanisms underlying individual proteins and explore proteomic clinical applications.

BACKGROUND Younger maternal age at birth is associated with increased risk of asthma in offspring in European descent populations, but has not been studied in Latino populations. OBJECTIVES We sought to examine the relationship between maternal age at birth and prevalence of asthma in a nationwide study of Latino children. METHODS We included 3473 Latino children aged 8-21 years (1696 subjects with physician-diagnosed asthma and 1777 healthy controls) from five US centres and Puerto Rico recruited from July 2008 through November 2011. We used multiple logistic regression models to examine the effect of maternal age at birth on asthma in offspring overall and in analyses stratified by ethnic subgroup (Mexican American, Puerto Rican and other Latino). Secondary analyses evaluated the effects of siblings, acculturation and income on this relationship. RESULTS Maternal age < 20 years was significantly associated with decreased odds of asthma in offspring, independent of other risk factors (OR = 0.73, 95% CI: 0.57-0.93). In subgroup analyses, the protective effect of younger maternal age was observed only in Mexican Americans (OR = 0.53, 95% CI: 0.36, 0.79). In Puerto Ricans, older maternal age was associated with decreased odds of asthma (OR = 0.65, 95% CI: 0.44-0.97). In further stratified models, the protective effect of younger maternal age in Mexican Americans was seen only in children without older siblings (OR = 0.44, 95% CI: 0.23-0.81). CONCLUSION AND CLINICAL RELEVANCE In contrast to European descent populations, younger maternal age was associated with decreased odds of asthma in offspring in Mexican American women. Asthma is common in urban minority populations but the factors underlying the varying prevalence among different Latino ethnicities in the United States is not well understood. Maternal age represents one factor that may help to explain this variability.

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Objective Managing staff stress and preventing long-term burnout in oncology staff are highly important for both staff and patient well-being. Research addressing work-related stress in adult oncology is well documented; however, less is known about this topic in the pediatric context. This study examined sources of work-related stress and reward specific to multidisciplinary staff working in pediatric oncology in Australia. Method Participants were 107 pediatric oncology clinicians, including medical, nursing, and allied health staff from two Australian pediatric oncology centers. Participants completed an online survey using two newly developed measures: the work stressors scale-pediatric oncology and the work rewards scale-pediatric oncology. Results The most commonly reported sources of both stress and reward are related to patient care and interactions with children. Results indicated that levels of work-related stress and reward were similar between the professional disciplines and between the two hospitals. Regression analyses revealed no demographic or organizational factors that were associated with either stress or reward. Conclusions Work-related stress and reward are not mutually exclusive; particular situations and events can be simultaneously stressful and rewarding for healthcare providers. Although patient care and interactions with children was found to be the most stressful aspect of working in this speciality, it was also the greatest source of reward. Results are discussed in relation to workplace approaches to staff well-being and stress reduction.

Background Very little research has examined the role of parenting in managing behavioural side-effects of cancer treatment. The purpose of this paper was to explore parent perceptions of (a) parenting in the context of childhood cancer; (b) the parenting strategies used in the context of managing child behavioural side-effects of cancer treatment; and (c) the perceived impact that cancer-specific parenting strategies have on child behaviour. Methods Participants were 15 mothers of children aged 2–6 years in the maintenance phase of treatment for acute lymphoblastic leukaemia at the Royal Children's Hospital Children's Cancer Centre, Melbourne, Australia. Mothers participated in a one-on-one semi-structured telephone interview using an interview guide which included questions on parenting in the context of childhood cancer, specifically in relation to behavioural side-effects (problems with behaviour, sleep and eating) and any perceived impact cancer-specific parenting may have on the ill child. Results Many parents reported that following their child's cancer diagnosis, they had to implement a suite of ‘new’ strategies that ‘pre-diagnosis’ were used only in moderation, if at all. The most salient theme that emerged was parents' perception that their parenting became more lax since their child's diagnosis. Parents further reported specific parenting strategies for each of the main child behavioural side-effects of cancer treatment. Conclusion Data from the current qualitative exploratory study highlight the role of specific parenting strategies in managing or assisting child behavioural side-effects of cancer treatment. Further quantitative research is needed to more fully examine the association between parenting and child behavioural outcomes in order to develop modifiable approaches to improving child behavioural side-effects in a paediatric oncology context.

Inhaled short-acting beta-agonist (SABA) medication is commonly used in asthma patients to rapidly reverse airway obstruction and improve acute symptoms. We performed a genome-wide association study of SABA medication response using gene-based association tests. A linear mixed model approach was first used for single-nucleotide polymorphism associations, and the results were later combined using GATES to generate gene-based associations. Our results identified SPATA13-AS1 as being significantly associated with SABA bronchodilator response in 328 healthy African Americans. In replication, this gene was associated with SABA response among the two separate groups of African Americans with asthma (n=1073, P=0.011 and n=1968, P=0.014), 149 healthy African Americans (P=0.003) and 556 European Americans with asthma (P=0.041). SPATA13-AS1 was also associated with longitudinal SABA medication usage in the two separate groups of African Americans with asthma (n=658, P=0.047 and n=1968, P=0.025). Future studies are needed to delineate the precise mechanism by which SPATA13-AS1 may influence SABA response.

Motivation: The inference of local ancestry of admixed individuals at every locus provides the basis for admixture mapping. Local ancestry information has been used to identify genetic susceptibility loci. Results: In this study, we developed a statistical method, efficient inference of local ancestry (EILA), which uses fused quantile regression and k-means classifier to infer the local ancestry for admixed individuals. We also conducted a simulation study using HapMap data to evaluate the performance of EILA in comparison with two competing methods, HAPMIX and LAMP. In general, the performance declined as the ancestral distance decreased and the time since admixture increased. EILA performed as well as the other two methods in terms of computational efficiency. In the case of closely related ancestral populations, all the three methods performed poorly. Most importantly, when the ancestral distance was large or moderate, EILA had higher accuracy and lower variation in comparison with the other two methods. Availability and implementation: EILA is implemented as an R package, which is freely available from the Comprehensive R Archive Network (http://cran.r-project.org/). Contact: moc.liamg@tatsgnayj

Managing staff stress and preventing long-term burnout in oncology staff are highly important for both staff and patient well-being. Research addressing work-related stress in adult oncology is well documented; however, less is known about this topic in the pediatric context. This study examined sources of work-related stress and reward specific to multidisciplinary staff working in pediatric oncology in Australia.Participants were 107 pediatric oncology clinicians, including medical, nursing, and allied health staff from two Australian pediatric oncology centers. Participants completed an online survey using two newly developed measures: the work stressors scale-pediatric oncology and the work rewards scale-pediatric oncology.The most commonly reported sources of both stress and reward are related to patient care and interactions with children. Results indicated that levels of work-related stress and reward were similar between the professional disciplines and between the two hospitals. Regression analyses revealed no demographic or organizational factors that were associated with either stress or reward.Work-related stress and reward are not mutually exclusive; particular situations and events can be simultaneously stressful and rewarding for healthcare providers. Although patient care and interactions with children was found to be the most stressful aspect of working in this speciality, it was also the greatest source of reward. Results are discussed in relation to workplace approaches to staff well-being and stress reduction.

BACKGROUND: Abnormal baseline renal function and worsening renal function (WRF) during heart failure (HF) hospitalization are associated with worse outcomes. However, which measure of renal function is the most prognostic is unknown. We sought to determine the renal function measure most predictive of in-hospital and post-discharge mortality. METHODS: This was a retrospective cohort study of patients with HF who received care from a large health system between Jan 1, 2000 and June 30, 2008. Patients with preexisting end-stage renal disease were excluded. Blood urea nitrogen (BUN), creatinine (Cr), and MDRD estimated glomerular filtration rate (eGFR) values at admission, and changes during hospitalization were tested for their association with in-hospital and one year post-discharge mortality. Logistic regression and conditional receiver operating curves for each index were used to determine the association with mortality by comparing the area under curve (AUC). RESULTS: Among 7394 patients meeting the inclusion criteria, 204 died in-hospital, and 1652 were discharged but died within 1 year. Baseline BUN was the strongest correlate for both in-hospital and post-discharge mortality (AUC= 0.724 and 0.656, respectively; ptable . CONCLUSIONS: Among hospitalized HF patients baseline BUN is the renal index most strongly associated with mortality. Common WRF definitions are suboptimal for predicting mortality, and the optimal thresholds depend on baseline renal function. Optimized WRF creatinine definition vs. traditional thresholds for predicting mortality (N=7394) BUN Strata ΔCreatinine cutpoint Sensitivity In-hospital death Specificity In-hospital death Odds Ratio 0 < BUN ≤ 15 0.2 0.3 0.5 94.4 (17/18) 72.2 (13/18) 50.0 (9/18) 48.9 (822/1680) 67.7 (1137/1680) 88.5 (1487/1680) 16.29 5.44 7.70 16 ≤ BUN ≤ 22 0.3 0.5 0.6 62.5 (20/32) 59.4 (19/32) 59.4 (19/32) 63.5 (1279/2014) 82.2 (1656/2014) 88.0 (1772/2014) 2.90 6.76 10.70 23 ≤ BUN ≤ 32 0.3 0.5 0.7 63.9 (23/36) 50.0 (18/36) 44.4 (16/36) 60.0 (1039/1731) 79.4 (1375/1731) 89.3 (1546/1731) 2.66 3.86 6.69 33 ≤ BUN < 200 0.3 0.5 0.8 67.8 (80/118) 60.2 (71/118) 54.9 (65/118) 56.9 (1005/1765) 74.2 (1310/1765) 85.8 (1514/1765) 2.78 4.35 7.40 BUN Strata ΔCreatinine cutpoint Sensitivity Post-discharge death Specificity Post-discharge death Odds ratio 0 < BUN ≤ 15 0.3 0.5 83.0 (181/218) 93.6 (204/218) 18.4 (206/1118) 7.4 (83/1118) 1.11 1.17 16 ≤ BUN ≤ 22 0.3 0.5 42.6 (142/333) 22.5 (75/333) 65.4 (902/1380) 83.7 (1155/1380) 1.40 1.49 23 ≤ BUN ≤ 32 0.1 0.3 0.5 73.3 (302/412) 56.6 (233/412) 20.9 (86/412) 33.0 (362/1096) 46.9 (514/1096) 80.1(878/1096) 1.35 1.15 1.06 33 ≤ BUN < 200 0.1 0.3 0.5 69.2 (477/689) 46.0 (317/689) 28.0 (193/689) 37.0 (331/895) 58.4 (523/895) 74.2 (664/895) 1.32 1.20 1.19

Fluorous phase soluble polymer supports derived from fluoroacrylate polymers are described. N-Acryloxysuccinimide-containing fluoroacrylate polymers were readily prepared from commercially available monomers. The activated acrylates so prepared were then converted into chelating and non-chelating ligands by amidation of the N-acryloxysuccinimide active ester residues. Phosphine ligands attached to these supports were used to prepare neutral and cationic rhodium(I) hydrogenation catalysts as well as palladium(0) catalysts. Similar substitution of pendant active ester groups to form hydroxamic acid ligands for metal sequestration is also feasible. Liquid/liquid extraction readily separated, recycled and reused these polymer-bound ligands and catalysts. While fluorous phase solubility could be attained with polymers containing only heptafluorobutyryl groups, selective solubility in a fluorous phase in contact with an organic phase was only seen with fluoroacrylates that contained larger fluorinated este r groups.

Sensitive individuals are at risk of adverse health effects from exposure to even modest levels of ozone. Such individuals are likely to develop respiratory symptoms and are more likely to be hospitalized when exposed to ozone at currently accepted levels. A review of scientific documents regarding the health effects of ozone has led the DHHS to agree with the EPA that the primary ambient air standard for ozone should be changed from 0.12 ppm (1-hour average) to 0.08 ppm (8-hour average). Achieving the proposed 0.08 ppm standard should decrease physician visits, hospitalization, and use of respiratory medications. In addition, because very susceptible healthy individuals may experience adverse health effects of the current standard, it is clear that the current standard provides little or no margin of safety, particularly for sensitive individuals. In order to avoid exposure to unhealthy levels of ozone, people should take precautions to minimize exposure from 2:00 to 6:00 pm on days when ozone levels are high. This is especially so for children, adults who are active outdoors, those with respiratory disease such as asthma, and people with unusual susceptibility to ozone.

This study examines perceptions of bankers regarding the usefulness of generally accepted accounting principles for small businesses financial statements. Perceptions were obtained regarding four accounting standards that have been criticized as being unnecessarily costly for smaller businesses. Although results were somewhat mixed, support was given to prior research that has suggested bankers would accept financial information prepared from a less costly non-GAAP basis for some small businesses.

The epidermal growth factor receptor (EGF-R) on human epidermoid carcinoma cells, A431, was found to be predominantly associated with the detergent-insoluble cytoskeleton, where it retained both a functional ligand-binding domain and an intrinsic tyrosine kinase activity. The EGF-R was constitutively associated with the A431 cytoskeleton; this association was not a consequence of adventitious binding. The EGF-R was associated with cytoskeletal elements both at the cell surface, within intracellular vesicles mediating the internalization of the hormone-receptor complex, and within lysosomes. The EGF-R became more stably associated with cytoskeletal elements after its internalization. The cytoskeletal association of the EGF-R was partially disrupted on suspension of adherent cells, indicating that alteration of cellular morphology influences the structural association of the EGF-R, and that the EGF-R is not intrinsically insoluble. Cytoskeletons prepared from EGF-treated A431 cells, when incubated with gamma-32P-ATP, demonstrated enhanced autophosphorylation of the EGF-R in situ as well as the phosphorylation of several high molecular weight proteins. In this system, phosphorylation occurs between immobilized kinase and substrate. The EGF-R and several high molecular weight cytoskeletal proteins were phosphorylated on tyrosine residues; two of the latter proteins were phosphorylated transiently as a consequence of EGF action, suggesting that EGF caused the active redistribution of the protein substrates relative to protein kinases. The ability of EGF to stimulate protein phosphorylation in situ required treatment of intact cells at physiological temperatures; addition of EGF directly to cytoskeletons had no effect. These data suggest that the structural association of the EGF-R may play a role in cellular processing of the hormone, as well as in regulation of the EGF-R kinase activity and in specifying its cellular substrates.

The binding of nerve growth factor (NGF) to specific cell surface receptors initiates a variety of effects that lead to the morphological and biochemical differentiation of clonal pheochromocytoma, PC12, cells. The lectin wheat germ agglutinin (WGA) alters the characteristics of NGF-receptor interaction. We have found that treatment of PC12 cells with WGA dramatically and reversibly inhibits the ability of NGF to elicit three distinct biological effects characteristic of NGF action. Two of these events, the rapid ruffling of cell-surface membranes and the stimulation of the phosphorylation of a 250-kD cytoskeletal protein in situ, occur rapidly and are an immediate consequence of receptor occupancy. Both of these effects are blocked by pretreatment of the cells with WGA. WGA was also found to inhibit the NGF-stimulated regeneration of neurites that occurs over 1-2 d. Both the WGA inhibition of neurite outgrowth and the phosphorylation of the 250-kD cytoskeletal protein were reversed upon addition of the specific sugar N-acetylglucosamine. These data demonstrate that the WGA-induced changes in the NGF-receptor interaction reflect important alterations in the ability of the receptor to transmit biological signals, resulting in the abrogation of the biological effects of NGF on these cells.

Monoclonal antibodies were raised against the human germ-cell tumor line, LICR LON HT39/7. The cells of this human embryonal carcinoma cell line were first fixed in formol saline and then dehydrated prior to immunization so that antibodies were produced which reacted with antigens surviving conventional histological processing. Eleven hybrid clones secreted antibody that reacted with both formol saline-fixed and living cells of the immunogen. One of these antibodies, LICR LON FC 10.2, has been found to react with a membrane glycoprotein of molecular weight 200,000 restricted in distribution to the surfaces of undifferentiated teratoma-derived cell lines, the luminal surface of human fetal intestine and bronchus and areas of putative endodermal differentiation within germ-cell neoplasms.

Three novel species-specific monoclonal antibodies 5.1.H, 8.7.D and 13.7.A raised against semi-purified detergent solubilized fractions of the Tera 1 embryonal carcinoma (EC) cell line are restricted in their in vitro distribution to undifferentiated human EC cell lines. Competition experiments have established that distinct antigenic specificities are seen by the 3 different monoclonal antibodies. All 3 antigens 5.1.H, 8.7.D and 13.7.A, defined by these monoclonal antibodies, undergo developmental regulation and cease to be expressed on Tera 2 clones 5 and 12 after retinoic-acid-induced differentiation and on LICR LON HT 39/7 cells after phorbol-ester-induced differentiation. These results taken together with the extremely limited in vivo tissue distribution of the defined antigens suggest that the 5.1.H, 8.7.D and 13.7.A monoclonal antibodies define distinct onco-foetal antigens.