A novel diagnostic gene region for distinguishing between two pest fruit flies: Bactrocera tryoni (Froggatt) and Bactrocera neohumeralis (Hardy) (Diptera: Tephritidae).

Bactrocera tryoni and Bactrocera neohumeralis are morphologically similar sibling pest fruit fly species that possess different biological attributes, geographic distributions, and host ranges. The need to differentiate between the two species is critical for accurate pest status assessment, management, biosecurity, and maintenance of reference colonies. While morphologically similar, adults may be separated based on subtle characters; however, some characters exhibit intraspecific variability, creating overlap between the two species. Additionally, there is currently no single molecular marker or rapid diagnostic assay that can reliably distinguish between B. neohumeralis and B. tryoni; therefore, ambiguous samples remain undiagnosed. Here we report the first molecular marker that can consistently distinguish between B. tryoni and B. neohumeralis. Our diagnostic region consists of two adjacent single nucleotide polymorphisms (SNPs) within the pangolin (pan) gene region. We confirmed the genotypes of each species are consistent across their distributional range, then developed a tetra-primer amplification refractory mutation system (ARMS) PCR assay for rapid diagnosis of the species. The assay utilizes four primers in multiplex, with two outer universal primers, and two internal primers: one designed to target two adjacent SNPs (AA) present in B. tryoni and the other targeting adjacent SNPs present in B. neohumeralis (GG). The assay accurately discriminates between the two species, but their SNP genotypes are shared with other nontarget tephritid fruit fly species. Therefore, this assay is most suited to adult diagnostics where species confirmation is necessary in determining ambiguous surveillance trap catches; maintaining pure colony lines; and in Sterile Insect Technique management responses.

Development of a cost-effective, morphology-preserving method for DNA isolation from bulk invertebrate trap catches: Tephritid fruit flies as an exemplar.

Insect identification and preservation of voucher specimens is integral to pest diagnostic and surveillance activities; yet bulk-trapped insects are a diagnostic challenge due to high catch numbers and the susceptibility of samples to environmental damage. Many insect trap catches rely on examination of morphological characters for species identifications, which is a time consuming and highly skilled task, hence there is a need for more efficient molecular approaches. Many bulk DNA extraction methods require destructive sampling of specimens, resulting in damaged, or fully destroyed, voucher specimens. We developed an inexpensive, rapid, bulk DNA isolation method that preserves specimens as pinned vouchers to a standard that allows for post-extraction morphological examination and inclusion in insect reference collections. Our protocol was validated using a group of insects that are time-consuming to identify when trapped in large numbers-the dacine fruit flies (Diptera: Tephritidae: Dacinae). In developing our method, we evaluated existing protocols against the following criteria: effect on morphology; suitability for large trap catches; cost; ease of handling; and application to downstream molecular diagnostic analyses such as real-time PCR and metabarcoding. We found that the optimum method for rapid isolation of DNA extraction was immersing flies in a NaOH:TE buffer at 75°C for 10 minutes, without the need for proteinase K or detergents. This HotSOAK method produced sufficient high-quality DNA whilst preserving morphological characters suitable for species-level identification with up to 20,000 flies in a sample. The lysates performed well in down-stream analyses such as loop-mediated isothermal amplification (LAMP) and real-time PCR applications, while for metabarcoding PCR the lysate required an additional column purification step. Development of this method is a key step required for upscaling our capacity to accurately detect insects captured in bulk traps, whether for biodiversity, biosecurity, or pest management objectives.

Exposure to the troubles in Northern Ireland, memory functioning, and social activity engagement: results from NICOLA.

We investigated the potential impact of a cohort traumatic exposure, the Troubles in Northern Ireland, on memory functioning in later life, and the potential moderating effect of social activity engagement. Using data from 6571 participants aged 60 + in the Northern Ireland Cohort for the Longitudinal Study of Ageing (NICOLA) cohort, we used a structural equation modelling framework to explore associations between traumatic exposure during the Troubles and memory functioning. As expected, social activity engagement was positively associated with memory functioning, ß = .102. Traumatic exposure was also positively associated with memory functioning, ß = .053. This association was stronger at low levels of social activity engagement; among those with higher levels, there was little association, interaction ß = - 0.054. The positive association between traumatic exposure during the Troubles and memory functioning was not moderated by the age at which the exposures occurred (based on analysis of a subsample with available data), interaction ß = - 0.015. We conclude that superior memory functioning was associated with higher levels of traumatic exposure during the Troubles, particularly among those with lower levels of social activity engagement, and regardless of the age at which the exposures occurred. Future longitudinal analyses are required to build on these results, which potentially have implications for life-course epidemiology, in relation to critical periods for traumatising experiences. Supplementary Information: The online version contains supplementary material available at 10.1007/s10433-022-00683-5.

Loop-mediated isothermal amplification (LAMP) assays for detection of the New Guinea fruit fly Bactrocera trivialis (Drew) (Diptera: Tephritidae).

The cue-lure-responding New Guinea fruit fly, Bactrocera trivialis, poses a biosecurity risk to neighbouring countries, e.g., Australia. In trapping programs, lure caught flies are usually morphologically discriminated from non-target species; however, DNA barcoding can be used to confirm similar species where morphology is inconclusive, e.g., Bactrocera breviaculeus and B. rufofuscula. This can take days-and a laboratory-to resolve. A quicker, simpler, molecular diagnostic assay would facilitate a more rapid detection and potential incursion response. We developed LAMP assays targeting cytochrome c oxidase subunit I (COI) and Eukaryotic Translation Initiation Factor 3 Subunit L (EIF3L); both assays detected B. trivialis within 25 min. The BtrivCOI and BtrivEIF3L assay anneal derivatives were 82.7 ± 0.8 °C and 83.3 ± 1.3 °C, respectively, detecting down to 1 × 101 copies/µL and 1 × 103 copies/µL, respectively. Each assay amplified some non-targets from our test panel; however notably, BtrivCOI eliminated all morphologically similar non-targets, and combined, the assays eliminated all non-targets. Double-stranded DNA gBlocks were developed as positive controls; anneal derivatives for the COI and EIF3L gBlocks were 84.1 ± 0.7 °C and 85.8 ± 0.2 °C, respectively. We recommend the BtrivCOI assay for confirmation of suspect cue-lure-trapped B. trivialis, with BtrivEIF3L used for secondary confirmation when required.

VATA-ADL: The Visual Analogue Test for Anosognosia for Activities of Daily Living.

OBJECTIVE: To study awareness of problems with one's own Activities of Daily Living (ADL) following stroke by means of a novel instrument-the Visual-Analogue Test for Anosognosia for Activities of Daily Living (VATA-ADL). METHODS: The new test overcomes some of the methodological problems of traditional structured interviews and self-rating questionnaires. In particular, to account for possible verbal communication difficulties, each question is illustrated by a drawing and a 4-point visual-analogue Likert scale. The patient's self-rating is compared with that given by informants (personal or professional caregiver) to acquire a measure of metacognition of one's own problems in performing everyday tasks. RESULTS: The VATA-ADL was validated in 61 dyads of older people and their informants. A group of 80 post-acute stroke patients and their informants then completed the test. Informant ratings correlated highly with traditional ADL scales, the questionnaire items showed high internal consistency (α = .95) and loaded onto one factor. By comparison to informants' assessments, the patients showed a generally poor appreciation of their functional disabilities. Thirty-nine patients overestimated their abilities (anosognosia) whereas nine showed underestimation of their abilities. CONCLUSIONS: Anosognosia (overestimation of abilities) for ADL is frequent, even in post-acute stages post-stroke. Some other patients underestimated their abilities, indicating that poor metacognition of one's own abilities in brain damaged patients is bi-directional. Both types of misestimation may have clinical consequences worth considering for the wellbeing of patients and their carers.

Conversation Initiation of Mothers, Fathers, and Toddlers in their Natural Home Environment.

In a conversational exchange, interlocutors use social cues including conversational turn-taking to communicate. There has been attention in the literature concerning how mothers, fathers, boys, and girls converse with each other, and in particular who initiates a conversation. Better understanding of conversational dynamics may deepen our understanding of social roles, speech and language development, and individual language variability. Here we use large-scale automatic analysis of 186 naturalistic daylong acoustic recordings to examine the conversational dynamics of 26 families with children about 30 months of age to better understand communication roles. Families included 15 with boys and 11 with girls. There was no difference in conversation initiation rate by child sex, but children initiated more conversations than mothers, and mothers initiated more than fathers. Results support developmental theories of the different and variable roles that interlocutors play in a social context.

A new brachylophosaurin (Dinosauria: Hadrosauridae) from the Upper Cretaceous Menefee Formation of New Mexico.

Brachylophosaurini is a clade of hadrosaurid dinosaurs from the Campanian of western North America. Although well-known from northern localities in Montana and Alberta, including abundant material of Brachylophosaurus canadensis and Maiasaura peeblesorum and the holotypes of Acristavus gagslarsoni and Probrachylophosaurus bergei, material from southern localities in Utah and Colorado is restricted to a partial skull referred to A. gagslarsoni and several indeterminate specimens. Here we describe Ornatops incantatus gen. et sp. nov., a new brachylophosaurin known from a partial skeleton from the Allison Member of the Menefee Formation in New Mexico. Ornatops is the first brachylophosaurin reported from New Mexico and the southernmost occurrence of the clade. Ornatops shares with Probrachylophosaurus and Brachylophosaurus a caudally expanded nasofrontal suture on the frontals, but also exhibits an autapomorphic nasofrontal suture morphology, with a horizontal rostral region and elevated caudal region with two prominent parasagittal bumps, which is different from other brachylophosaurin specimens, including juvenile and adult Brachylophosaurus. A phylogenetic analysis places Ornatops in a trichotomy with Probrachylophosaurus and Brachylophosaurus, with Maiasaura and Acristavus as successive outgroups.

Transitional evolutionary forms in chasmosaurine ceratopsid dinosaurs: evidence from the Campanian of New Mexico.

Three new chasmosaurines from the Kirtland Formation (~75.0-73.4 Ma), New Mexico, form morphological and stratigraphic intermediates between Pentaceratops (~74.7-75 Ma, Fruitland Formation, New Mexico) and Anchiceratops (~72-71 Ma, Horseshoe Canyon Formation, Alberta). The new specimens exhibit gradual enclosure of the parietal embayment that characterizes Pentaceratops, providing support for the phylogenetic hypothesis that Pentaceratops and Anchiceratops are closely related. This stepwise change of morphologic characters observed in chasmosaurine taxa that do not overlap stratigraphically is supportive of evolution by anagenesis. Recently published hypotheses that place Pentaceratops and Anchiceratops into separate clades are not supported. This phylogenetic relationship demonstrates unrestricted movement of large-bodied taxa between hitherto purported northern and southern provinces in the late Campanian, weakening support for the hypothesis of extreme faunal provincialism in the Late Cretaceous Western Interior.

Scientific Review Committees as part of institutional review of human participant research: Initial implementation at institutions with Clinical and Translational Science Awards.

INTRODUCTION: Scientific quality and feasibility are part of ethics review by Institutional Review Boards (IRBs). Scientific Review Committees (SRCs) were proposed to facilitate this assessment by the Clinical and Translational Science Award (CTSA) SRC Consensus Group. This study assessed SRC feasibility and impact at CTSA-affiliated academic health centers (AHCs). METHODS: SRC implementation at 10 AHCs was assessed pre/post-intervention using quantitative and qualitative methods. Pre-intervention, four AHCs had no SRC, and six had at least one SRC needing modifications to better align with Consensus Group recommendations. RESULTS: Facilitators of successful SRC implementation included broad-based communication, an external motivator, senior-level support, and committed SRC reviewers. Barriers included limited resources and staffing, variable local mandates, limited SRC authority, lack of anticipated benefit, and operational challenges. Research protocol quality did not differ significantly between study periods, but respondents suggested positive effects. During intervention, median total review duration did not lengthen for the 40% of protocols approved within 3 weeks. For the 60% under review after 3 weeks, review was lengthened primarily due to longer IRB review for SRC-reviewed protocols. Site interviews recommended designing locally effective SRC processes, building buy-in by communication or by mandate, allowing time for planning and sharing best practices, and connecting SRC and IRB procedures. CONCLUSIONS: The CTSA SRC Consensus Group recommendations appear feasible. Although not conclusive in this relatively short initial implementation, sites perceived positive impact by SRCs on study quality. Optimal benefit will require local or federal mandate for implementation, adapting processes to local contexts, and employing SRC stipulations.

Building On The Gains Of The ACA: Federal Proposals To Improve Coverage And Affordability.

The Affordable Care Act (ACA) significantly improved health insurance coverage in the US, but too many Americans remain under- or uninsured. This article examines federal strategies under consideration that build on the ACA to extend comprehensive coverage to all low-income Americans and increase coverage affordability for middle-income Americans. For low-income Americans these policy options include extending the enhanced match rate offered to states that expanded eligibility for Medicaid in the early years of the ACA to states that have not yet expanded Medicaid and increasing Marketplace cost-sharing subsidies. To address the issue of affordability for middle-income Americans, this article considers options for lowering premiums (for example, extending tax credits to people with incomes above the current eligibility threshold, increasing the generosity of tax credits for those currently eligible, and making reinsurance permanent), lowering cost sharing (such as tying premium tax credits to the second-lowest-cost gold plan rather than the equivalent silver plan and extending federal assistance for cost sharing to people with incomes above the current threshold), and establishing a public option.

Pan-TRK Immunohistochemistry: A Useful Diagnostic Adjunct For Secretory Carcinoma of the Breast.

Secretory carcinoma is a special-type breast carcinoma underpinned by a recurrent t(12;15)(p13;q25) translocation resulting in ETV6-NTRK3 gene fusion. Immunohistochemistry (IHC) using a pan-TRK antibody has been recently shown to help identify NTRK rearrangements in other tumor types. The purpose of this study was to assess the diagnostic utility of pan-TRK IHC in secretory carcinoma of the breast. Pan-TRK IHC was performed using a rabbit monoclonal antibody on whole sections of 24 breast secretory carcinomas and tissue microarray sections of other breast carcinoma types (n=203) and histologic mimics (n=15). Cases were assessed for staining intensity and localization. The 24 patients with secretory carcinoma had a median age of 44 years and a median tumor size of 1.0 cm. ETV6 fluorescence in situ hybridization was positive in all cases tested (n=20). Twenty-three cases (95.8%) showed staining with pan-TRK, which was exclusively nuclear in 19, primarily nuclear with weak cytoplasmic staining in 3, and primarily cytoplasmic with focal nuclear staining in 1. The nuclear staining was diffuse in 17 and at least focally strong in 17. The only pan-TRK negative case was a core biopsy with limited tumor. Among the 203 nonsecretory carcinomas, 21 (10.3%) showed focal, weak nuclear staining in <5% of tumor cells and 1 (0.5%) showed focal membranous staining. All histologic mimics were negative. In conclusion, diffuse and/or at least focally strong nuclear pan-TRK staining is a sensitive and specific marker for secretory carcinoma of the breast.

Wise up: Clarifying the role of metacognition in the Dunning-Kruger effect.

The Dunning-Kruger effect (DKE) is the finding that, across a wide range of tasks, poor performers greatly overestimate their ability, whereas top performers make more accurate self-assessments. The original account of the DKE involves the idea that metacognitive insight requires the same skills as task performance, so that unskilled people perform poorly and lack insight. However, global measures of self-assessment are prone to statistical and other biases that could explain the same pattern. We used psychophysical methods to examine metacognitive insight in simple movement and spatial memory tasks: pointing at a dot or recalling its position after a delay. We measured task skill in an initial block, and self-assessment in a second block, in which participants judged after every trial whether they had hit the target or not. Metacognitive calibration and sensitivity were related to task skill, but a path analysis showed that their net contribution to the DKE was weak. The major driver of the DKE was the level of task performance. In a second study, we again measured task skill in an initial block, but titrated task difficulty in the second block so that all participants performed at equivalent levels of success. Metacognitive measures were again related to task skill, but the DKE pattern was eliminated. We present a simple model of these findings, showing that metacognitive differences can contribute to the DKE but are neither necessary nor sufficient for it. This analysis clarifies and quantifies how metacognitive insight and other factors interact to determine this famous effect. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

Over- and underestimation of motor ability after a stroke: Implications for anosognosia.

We administered a discrepancy-based measure of anosognosia for hemiplegia (AHP) to a group of 42 right-brain-damaged (RBD) and left-brain-damaged (LBD) stroke patients with varying levels of functional motor ability. In addition to the expected (anosognosic) pattern of overestimation of motor function in some RBD patients, we found an equal and opposite underestimation in some others, both RBD and LBD. We also found that around a quarter of self-estimation error could be predicted directly from actual ability, such that patients with poorer motor function tended to overestimate, and vice versa. This pattern suggests that some misestimation is attributable simply to statistical regression. However, even after adjusting for this regression effect, levels of overestimation were significantly greater in RBD patients, while LBD patients were more likely to underestimate their motor ability.

In Vivo Characterisation of Five Strains of Bovine Viral Diarrhoea Virus 1 (Subgenotype 1c).

Bovine viral diarrhoea virus 1 (BVDV-1) is strongly associated with several important diseases of cattle, such as bovine respiratory disease, diarrhoea and haemoragic lesions. To date many subgenotypes have been reported for BVDV-1, currently ranging from subgenotype 1a to subgenotype 1u. While BVDV-1 has a world-wide distribution, the subgenotypes have a more restricted geographical distribution. As an example, BVDV-1 subgenotypes 1a and 1b are frequently detected in North America and Europe, while the subgenotype 1c is rarely detected. In contrast, BVDV-1 subgenotype 1c is by far the most commonly reported in Australia. Despite this, uneven distribution of the biological importance of the subgenotypes remains unclear. The aim of this study was to characterise the in vivo properties of five strains of BVDV-1 subgenotype 1c in cattle infection studies. No overt respiratory signs were reported in any of the infected cattle regardless of strain. Consistent with other subgenotypes, transient pyrexia and leukopenia were commonly identified, while thrombocytopenia was not. The quantity of virus detected in the nasal secretions of transiently infected animals suggested the likelihood of horizontal transmission was very low. Further studies are required to fully understand the variability and importance of the BVDV-1 subgenotype 1c.

Acquired progressive kinking of the hair in an elderly woman: a case report.

Acquired progressive kinking of the hair (APK) is a rare disorder that presents at any age and is infrequently presented in the literature. This diagnosis has been used to describe similar hair changes, though there are significant differences in presentation, population, and individual clinical features. Histologic analysis of hair shafts aids in diagnosis owing to commonly reported features such as irregular twisting, bending, and grooving of hair shafts with polygonal shapes on cross-section. We present a case with classic histologic features in an unusual patient to highlight this rare condition. Additional cases and studies are necessary to evaluate etiology, differentiate subsets of APK, and develop both treatment and prevention strategies.

Altered Expression of Angiotensinogen and Mediators of Angiogenesis in Ileal Crohn's Disease.

BACKGROUND AND AIMS: Angiotensin II (AII) is a powerful splanchnic vasoconstrictor with pro-inflammatory and pro-fibrotic properties. Angiotensin converting enzyme (ACE) inhibitors and AII Receptor Antagonists (ARBs) are therapeutic in animal models of colitis. The aim of this case-control study is to determine the expression of angiotensinogen and related genes in human ileal Crohn's disease. METHODS: Using quantitative real-time polymerase chain reaction (RT-PCR), we measured mRNA expression levels of angiotensinogen (AGT), hypoxia inducible factor (HIF)1α and melanoma cell adhesion molecule (MCAM; CD146) in 101 human samples (69 biopsy, 12 resection) from affected ileum (inflamed CD cases, n=36) and unaffected ileum (non-inflamed CD cases, n=45 and controls, n=20). Immunohistochemistry for angiotensinogen was also performed. The study was of case-control design in a tertiary care setting. RESULTS: Ileal expression of AGT was lower in CD cases compared to controls (p<0.0001), in inflamed CD samples (p=0.017) and current smokers (p=0.02). HIF1α expression was lower in non-inflamed CD biopsy samples than controls (p=0.006). The presence of disease-associated NOD2 variants was associated with increased expression of markers of angiogenesis (HIF1α p=0.009; MCAM p=0.007) in inflamed CD samples. Angiotensinogen immunohistochemical staining supported the quantitative RT-PCR expression findings. CONCLUSIONS: Angiotensinogen expression is down regulated in human ileal CD, particularly in the presence of inflammation and current cigarette smoking, implicating the mesenteric vasculature and mucosal hypoxia as co-factors in ileal CD pathogenesis. A novel reduction in HIF1α expression in non-inflamed ileal mucosa in CD patients was also demonstrated.

The first genome sequence of a metatherian herpesvirus: Macropodid herpesvirus 1.

BACKGROUND: While many placental herpesvirus genomes have been fully sequenced, the complete genome of a marsupial herpesvirus has not been described. Here we present the first genome sequence of a metatherian herpesvirus, Macropodid herpesvirus 1 (MaHV-1). RESULTS: The MaHV-1 viral genome was sequenced using an Illumina MiSeq sequencer, de novo assembly was performed and the genome was annotated. The MaHV-1 genome was 140 kbp in length and clustered phylogenetically with the primate simplexviruses, sharing 67% nucleotide sequence identity with Human herpesviruses 1 and 2. The MaHV-1 genome contained 66 predicted open reading frames (ORFs) homologous to those in other herpesvirus genomes, but lacked homologues of UL3, UL4, UL56 and glycoprotein J. This is the first alphaherpesvirus genome that has been found to lack the UL3 and UL4 homologues. We identified six novel ORFs and confirmed their transcription by RT-PCR. CONCLUSIONS: This is the first genome sequence of a herpesvirus that infects metatherians, a taxonomically unique mammalian clade. Members of the Simplexvirus genus are remarkably conserved, so the absence of ORFs otherwise retained in eutherian and avian alphaherpesviruses contributes to our understanding of the Alphaherpesvirinae. Further study of metatherian herpesvirus genetics and pathogenesis provides a unique approach to understanding herpesvirus-mammalian interactions.