Polarised cell intercalation during Drosophila axis extension is robust to an orthogonal pull by the invaginating mesoderm.

As tissues grow and change shape during animal development, they physically pull and push on each other, and these mechanical interactions can be important for morphogenesis. During Drosophila gastrulation, mesoderm invagination temporally overlaps with the convergence and extension of the ectodermal germband; the latter is caused primarily by Myosin II-driven polarised cell intercalation. Here, we investigate the impact of mesoderm invagination on ectoderm extension, examining possible mechanical and mechanotransductive effects on Myosin II recruitment and polarised cell intercalation. We find that the germband ectoderm is deformed by the mesoderm pulling in the orthogonal direction to germband extension (GBE), showing mechanical coupling between these tissues. However, we do not find a significant change in Myosin II planar polarisation in response to mesoderm invagination, nor in the rate of junction shrinkage leading to neighbour exchange events. We conclude that the main cellular mechanism of axis extension, polarised cell intercalation, is robust to the mesoderm invagination pull. We find, however, that mesoderm invagination slows down the rate of anterior-posterior cell elongation that contributes to axis extension, counteracting the tension from the endoderm invagination, which pulls along the direction of GBE.

Different temporal requirements for tartan and wingless in the formation of contractile interfaces at compartmental boundaries.

Compartmental boundaries physically separate developing tissues into distinct regions, which is fundamental for the organisation of the body plan in both insects and vertebrates. In many examples, this physical segregation is caused by a regulated increase in contractility of the actomyosin cortex at boundary cell-cell interfaces, a property important in developmental morphogenesis beyond compartmental boundary formation. We performed an unbiased screening approach to identify cell surface receptors required for actomyosin enrichment and polarisation at parasegmental boundaries (PSBs) in early Drosophila embryos, from the start of germband extension at gastrulation and throughout the germband extended stages (stages 6 to 11). First, we find that Tartan is required during germband extension for actomyosin enrichment at PSBs, confirming an earlier report. Next, by following in real time the dynamics of loss of boundary straightness in tartan mutant embryos compared with wild-type and ftz mutant embryos, we show that Tartan is required during germband extension but not beyond. We identify candidate genes that could take over from Tartan at PSBs and confirm that at germband extended stages, actomyosin enrichment at PSBs requires Wingless signalling.

An evaluation of 4 commercial assays for the detection of SARS-CoV-2 antibodies in a predominantly mildly symptomatic low prevalence Australian population.

A total of 1080 individual patient samples (158 positive serology samples from confirmed, predominantly mildly symptomatic COVID-19 patients and 922 serology negative including 496 collected pre-COVID) from four states in Australia were analysed on four commercial SARS-CoV-2 serological assays targeting antibodies to different antigens (Roche Elecsys and Abbott Architect: nucleocapsid; Diasorin Liaison and Euroimmun: spike). A subset was compared to immunofluorescent antibody (IFA) and micro-neutralisation. Sensitivity and specificity of the Roche (n = 1033), Abbott (n = 806), Diasorin (n = 1034) and Euroimmun (n = 175) were 93.7 %/99.5 %, 90.2 %/99.4 %, 88.6 %/98.6 % and 91.3 %/98.8 %, respectively. ROC analysis with specificity held at 99 % increased the sensitivity for the Roche and Abbott assays from 93.7% to 98.7% (cut-off 0.21) and 90.2 % to 94.0 % (cut-off 0.91), respectively. Overall seropositivity of samples increased from a maximum of 23 % for samples 0-7 days-post-onset of symptoms (dpos), to 61 % from samples 8-14dpos and 93 % from those >14dpos. IFA and microneutralisation values correlated best with assays targeting antibodies to spike protein with values >80 AU/mL on the Diasorin assay associated with neutralising antibody. Detectable antibody was present in 22/23 (96 %), 20/23 (87 %), 15/23 (65 %) and 9/22 (41 %) patients with samples >180dpos on the Roche, Diasorin, Abbott and microneutralisation assays respectively. Given the low prevalence in this community, two-step algorithms on initial positive results saw an increase in the positive predictive value (PPV) of positive samples (39 %-65 % to ≥98 %) for all combinations. Similarly accuracy increased from a range of 98.5 %-99.4 % to ≥99.8 % assuming a 1 % seroprevalence. Negative predictive value (NPV) was high (≥99.8 %) regardless of which assay was used initially.

Pertussis toxin IgA testing over-diagnoses recent pertussis infection.

The importance of pertussis toxin (PT) IgA testing in the diagnosis of recent pertussis infection remains unclear. The contribution of PT IgA to the diagnosis of recent pertussis was reviewed in two separate analyses. Firstly, an evaluation of two new automated assays [DiaSorin Liaison (DL), Italy] for PT IgG and PT IgA provided an opportunity to assess the contribution of PT IgA testing to PT IgG results. Secondly, a retrospective review of results from the PT IgA assay currently in use [Sullivan Nicolaides Pathology (SNP) PT IgA] was performed from 2013 to 2015 (n=63,474). For both the DL and SNP assays, the combination of PT IgG and PT IgA resulted in reduced specificity as compared to PT IgG results alone. For DL assays, an algorithm restricting DL PT IgA testing to samples with equivocal PT IgG results, demonstrated superior specificity to routinely testing both assays. The retrospective review indicated that only a minority of patients had a SNP PT IgA response without an accompanying rise in SNP PT IgG. There was also evidence of an age-related increase in the prevalence of isolated positive SNP PT IgA results which did not appear to be associated with recent pertussis infection. In general, PT IgA appears to contribute little diagnostic value to an accurate PT IgG assay in a community-based, Australian population. Reflex testing of PT IgA in the context of equivocal PT IgG results may be worthwhile if laboratory workflow permits.

The Cost-Effectiveness of Intensive Interdisciplinary Pediatric Chronic Pain Rehabilitation.

OBJECTIVE: Examine the cost-effectiveness of a 3-week interdisciplinary pediatric chronic pain rehabilitation program. METHODS: Self-reported health care utilization and parent missed work of youth with chronic pain (n = 127) at admission and 1-year follow-up were compared. Financials were calculated from program revenue and established national costs for health care and wages. RESULTS: Data indicate significant reductions in days hospitalized, physician office visits, physical/occupational therapy services, psychotherapy visits, and parental missed work. Estimated health care expenses were $61,988 in the year before admission and $14,189 in the year after admission (-$58,839). Estimated cost of missed work was $12,229 in the year prior and $1,189 in the year after (-$11,039). CONCLUSIONS: Comparing estimated expenses before ($74,217) and after ($15,378) minus program costs ($31,720), yielded estimated savings of $27,119 per family in the year following admission. These findings extend the benefit of the program beyond clinical improvement, to outcomes important to both families and insurers.

Psychometric Properties of the Pain Stages of Change Questionnaire: New Insights on the Measurement of Readiness to Change in Adolescents, Mothers, and Fathers.

UNLABELLED: There is increasing interest in the measurement of "readiness to change," or willingness to engage in a self-management approach to pain coping, as a predictor of treatment response in pediatric pain populations. The primary aim of the present study was to provide cross-validation of the Pain Stages of Change Questionnaire-Adolescent and -Parent versions in a new, independent pediatric chronic pain sample by examining aspects of reliability, validity, and generalizability of the factor structures identified in the initial validation study. Secondary aims were to 1) expand upon previously identified differences between the Pain Stages of Change Questionnaire-Adolescent and -Parent versions and 2) examine previously unreported aspects of father data. Although slight differences emerged, the factor structures identified in the initial validation were largely replicated, suggesting that the psychometric properties of the measure are robust across pediatric outpatient chronic pain samples. Variability between parent and adolescent reports suggests that there may be meaningful differences in the interpretation of each measure and that factors other than readiness to change may influence response patterns. Findings highlight the need for more fine-tuned analyses of the way the construct operates in youth with pediatric pain and their parents. PERSPECTIVE: Findings provide further validation of the Pain Stages of Change Questionnaire-Adolescent and -Parent versions measures in a new outpatient pediatric chronic pain sample. Previously uninvestigated father data showed good reliability and patterns of findings similar to validated mother reports. Moreover, the study suggests that the adolescent and parent versions may function in meaningfully different ways.

Developing an Alternative Rorschach Administration Method to Optimize the Number of Responses and Enhance Clinical Inferences.

UNLABELLED: Variability in the number of Rorschach responses (R) has stimulated controversy among clinicians and researchers for many years, and recent research reveals that R is much more variable than previously thought. Because R is correlated with other scores, its excessive variability may reduce the reliability, validity and clinical utility of these other scores. We present two experimental studies and additional results from other clinical datasets with the aim of developing a new administration procedure to diminish variability in R by reducing the number of very short and long records. In the first experiment, protocols were obtained using standard Comprehensive System administration or an alternative where we encouraged a second response if only one was given to a card and allowed only four responses on each card. This alternative method reduced the proportion of short records but produced an undesirable number of long records. To minimize the proportion of long records, in a second experiment, we added an instruction to give two or maybe three responses per card when introducing the test. Comparisons to CS administrations revealed that this procedure reduced variability in R by limiting the proportion of both short and long records. This reduced range was largely retained in an outpatient sample of older respondents with schizophrenia and a mixed clinical sample. Thus, we recommend this method of optimizing the range of R, which has since been included with very minor changes in the Rorschach Performance Assessment System. KEY PRACTITIONER MESSAGE: Alternative administration method successfully eliminated overly short and excessively long records. Utility is potentially increased by greatly reducing both short records that often lack reliability and validity, as well as long records that consume an excessive amount of examiner administration and scoring time. Psychometric properties and the ability to apply parametric statistics are likely increased across variables given that the distribution of R is more normal. Re-administration due to inadequate R is almost never needed. Results are consistent with the conclusion that this alternative procedure reduces examiner variability by offering simple, but explicit instructions for encouraging sufficient productivity. Overall variability of R produced using the refined alternative procedure was significantly less than that produced using the traditional CS method, although more in line with Exner's (2003) normative expectations. Suggests that when using the alternative method, R becomes less of a confound for all other scores that are moderately to highly correlated with R. Also demonstrated that the reduced variability of R and the reduced number of less useful short and long records are generalized to clinical samples. Additional research (Reese, Viglione, & Giromini, 2014) provides support for these conclusions with child clinical samples.

The impact of disability on the lives of children; cross-sectional data including 8,900 children with disabilities and 898,834 children without disabilities across 30 countries.

BACKGROUND: Children with disabilities are widely believed to be less likely to attend school or access health care, and more vulnerable to poverty. There is currently little large-scale or internationally comparable evidence to support these claims. The aim of this study was to investigate the impact of disability on the lives of children sponsored by Plan International across 30 countries. METHODS AND FINDINGS: We conducted a cross-sectional survey including 907,734 children aged 0-17 participating in the Plan International Sponsorship Programme across 30 countries in 2012. Parents/guardians were interviewed using standardised questionnaires including information on: age, sex, health, education, poverty, and water and sanitation facilities. Disability was assessed through a single question and information was collected on type of impairment. The dataset included 8,900 children with reported disabilities across 30 countries. The prevalence of disability ranged from 0.4%-3.0% and was higher in boys than girls in 22 of the 30 countries assessed - generally in the range of 1.3-1.4 fold higher. Children with disabilities were much less likely to attend formal education in comparison to children without disabilities in each of the 30 countries, with age-sex adjusted odds ratios exceeding 10 for nearly half of the countries. This relationship varied by impairment type. Among those attending school, children with disabilities were at a lower level of schooling for their age compared to children without disabilities. Children with disabilities were more likely to report experiencing a serious illness in the last 12 months, except in Niger. There was no clear relationship between disability and poverty. CONCLUSIONS: Children with disabilities are at risk of not fulfilling their educational potential and are more vulnerable to serious illness. This exclusion is likely to have a long-term deleterious impact on their lives unless services are adapted to promote their inclusion.

Development and validation of an Australian in-house anti-pertussis toxin IgG and IgA enzyme immunoassay.

AIMS: Although anti-pertussis toxin (PT) immunoglobulin G (IgG) is considered one of the most specific serological markers for Bordetella pertussis infection, there are few commercial kits available in Australia. We aimed to present the process of development, quality control and on-going clinical validation of an anti-PT IgG and IgA enzyme immunoassay (EIA) in use since 1999, and discuss the application of such tests in the diagnosis of B. pertussis infections. METHODS: A total of 1311 serum samples were used during multiple clinical validations from 1998 to 2010. The samples were drawn from healthy adults, children, patients with other respiratory infections, and patients with confirmed pertussis. Assay reproducibility, accuracy and precision criteria conformed to National Pathology Accreditation Advisory Council (NPAAC) guidelines. RESULTS: Using the World Health Organization clinical and/or laboratory definition of a definite case as the comparative standard, sensitivity was 84% [95% confidence interval (CI) 75-93] and specificity was 98% (95%CI: 90-100) for anti-PT IgG. Sensitivity was 72% (95%CI 64-80) and specificity was 98% (95%CI 90-100) for anti-PT IgA. There was minimal background positivity in either healthy adults or children using the established cut-offs. There was no appreciable effect of immunisation or cross reactions with other respiratory pathogens. CONCLUSION: Serological evaluation of various populations enabled the development of an anti-PT IgG and IgA EIA assay which was suitable for the diagnosis of acute infection in convalescent samples from clinically confirmed cases. Repeated evaluations of population-based cut-offs are required for in-house assays to ensure they remain clinically relevant. The subsequent validation of the cut-offs with WHO international standards has been published in a recent prospective study.

The psychometric properties of the screen for child anxiety related emotional disorders in pediatric chronic pain.

OBJECTIVE: Examine the psychometric properties of the SCARED in pediatric chronic pain. METHODS: Participants were parents (n = 313 mothers, 163 fathers) and youth (n = 349) presenting for treatment of pediatric chronic pain. Participants completed the SCARED and measures of pain catastrophizing, internalizing problems, and health-related quality of life. RESULTS: Internal consistency (Cronbach's α) of SCARED Total scores ranged from .92 to .93 across sources of report. All subscales except for School Phobia exhibited good internal consistency. SCARED scores were significantly positively related to internalizing symptoms and pain catastrophizing; and negatively related to health-related quality of life. Confirmatory factor analysis revealed acceptable fit of the SCARED measurement model. CONCLUSIONS: The SCARED shows promise as a measure of anxiety in pediatric pain. Important caveats for its usage and areas in need of additional research are discussed. Of importance in pediatric pain is improving current approaches for measuring school anxiety in this population.

Prospective evaluation of an Australian pertussis toxin IgG and IgA enzyme immunoassay.

Serological diagnosis of recent pertussis infection is an important part of both clinical assessment and epidemiological documentation of this disease. Standardization of serological testing and interpretation remains challenging despite international efforts to improve it. Currently, determining the anti-pertussis toxin (PT) IgG titer is recommended as the most accurate serological test in Europe and the United States, while Australia relies predominantly on measurement of Bordetella pertussis IgA antibody responses. Using B. pertussis PCR and the WHO clinical case definition as reference standards, the diagnostic utility of in-house anti-PT IgG and anti-PT IgA assays was evaluated prospectively in an Australian community-based cohort (n = 327). Patients provided up to four consecutive serum samples to document the kinetics of antibody response and decay. Previously validated cutoffs for positivity were converted to international units by using WHO-approved reference sera. At currently used cutoffs, both anti-PT IgG (>94 IU/ml) and anti-PT IgA (>20 IU/ml) assays had good specificity (80% [95% confidence interval {95% CI}, 68 to 88%] and 87% [95% CI, 77 to 94%]), but anti-PT IgG assay was consistently more sensitive than anti-PT IgA assay across a range of cutoffs (60 to 79% [95% CI, 53 to 84%] versus 41 to 62% [95% CI, 34 to 69%]). The combination of anti-PT IgG and anti-PT IgA assays performed no better than anti-PT IgG assay alone. The anti-PT IgA response in children under 12 years of age was poor. The accuracy of serology was optimal between 2 and 8 weeks after symptom onset. Cutoffs of >94 IU/ml for anti-PT IgG and >20 IU/ml for anti-PT IgA correlated well with recent pertussis infection and were consistent with recent recommendations from the EU Pertstrain group. Anti-PT IgG assay was superior to anti-PT IgA assay as the test of choice for the diagnosis of pertussis from a single sample.