A genome-wide screen identifies SCAI as a modulator of the UV-induced replicative stress response.

Helix-destabilizing DNA lesions induced by environmental mutagens such as UV light cause genomic instability by strongly blocking the progression of DNA replication forks (RFs). At blocked RF, single-stranded DNA (ssDNA) accumulates and is rapidly bound by Replication Protein A (RPA) complexes. Such stretches of RPA-ssDNA constitute platforms for recruitment/activation of critical factors that promote DNA synthesis restart. However, during periods of severe replicative stress, RPA availability may become limiting due to inordinate sequestration of this multifunctional complex on ssDNA, thereby negatively impacting multiple vital RPA-dependent processes. Here, we performed a genome-wide screen to identify factors that restrict the accumulation of RPA-ssDNA during UV-induced replicative stress. While this approach revealed some expected "hits" acting in pathways such as nucleotide excision repair, translesion DNA synthesis, and the intra-S phase checkpoint, it also identified SCAI, whose role in the replicative stress response was previously unappreciated. Upon UV exposure, SCAI knock-down caused elevated accumulation of RPA-ssDNA during S phase, accompanied by reduced cell survival and compromised RF progression. These effects were independent of the previously reported role of SCAI in 53BP1-dependent DNA double-strand break repair. We also found that SCAI is recruited to UV-damaged chromatin and that its depletion promotes nascent DNA degradation at stalled RF. Finally, we (i) provide evidence that EXO1 is the major nuclease underlying ssDNA formation and DNA replication defects in SCAI knockout cells and, consistent with this, (ii) demonstrate that SCAI inhibits EXO1 activity on a ssDNA gap in vitro. Taken together, our data establish SCAI as a novel regulator of the UV-induced replicative stress response in human cells.

The Nrd1-like protein Seb1 coordinates cotranscriptional 3' end processing and polyadenylation site selection.

Termination of RNA polymerase II (RNAPII) transcription is associated with RNA 3' end formation. For coding genes, termination is initiated by the cleavage/polyadenylation machinery. In contrast, a majority of noncoding transcription events in Saccharomyces cerevisiae does not rely on RNA cleavage for termination but instead terminates via a pathway that requires the Nrd1-Nab3-Sen1 (NNS) complex. Here we show that the Schizosaccharomyces pombe ortholog of Nrd1, Seb1, does not function in NNS-like termination but promotes polyadenylation site selection of coding and noncoding genes. We found that Seb1 associates with 3' end processing factors, is enriched at the 3' end of genes, and binds RNA motifs downstream from cleavage sites. Importantly, a deficiency in Seb1 resulted in widespread changes in 3' untranslated region (UTR) length as a consequence of increased alternative polyadenylation. Given that Seb1 levels affected the recruitment of conserved 3' end processing factors, our findings indicate that the conserved RNA-binding protein Seb1 cotranscriptionally controls alternative polyadenylation.

Interrater reliability of the Standing and Walking Assessment Tool for spinal cord injury.

STUDY DESIGN: Psychometric study. OBJECTIVES: The Standing and Walking Assessment Tool (SWAT) is a standardized approach to the evaluation of standing and walking capacity following traumatic spinal cord injury (tSCI) in Canada. The SWAT classifies individuals with a tSCI into 12 stages of standing and walking capacity that are paired with well-established outcome measures, such as the Berg Balance Scale and 10-m Walk Test. Prior research has demonstrated the validity and responsiveness of the SWAT stages; however, the reliability remains unknown. The objective of this study was to evaluate the interrater reliability of the SWAT stages. SETTING: Inpatient units of two Canadian rehabilitation hospitals. METHODS: Adults with sub-acute tSCI were recruited. SWAT stage was evaluated for each participant by two physical therapists separately. The two therapists aimed to complete the evaluations within one day of each other. To evaluate interrater reliability, the percentage agreement between the SWAT stages rated by the two physical therapists was calculated, along with a linear weighted kappa statistic with a 95% CI. RESULTS: Forty-five individuals with sub-acute tSCI (36 males, 9 females, mean (SD) age of 54.8 (17.9) years) participated. The percentage agreement in SWAT stages between the two physical therapists was 75.6%. A kappa statistic of 0.93 with a 95% CI, 0.81-1.05 was obtained. In cases where therapists disagreed (18% of participants), therapists differed by 1-2 stages only. CONCLUSIONS: The SWAT stages have high interrater reliability, providing further support for the use of the SWAT in rehabilitation practice in Canada.

Bioengineering of the Marine Diatom Phaeodactylum tricornutum with Cannabis Genes Enables the Production of the Cannabinoid Precursor, Olivetolic Acid.

The increasing demand for novel natural compounds has prompted the exploration of innovative approaches in bioengineering. This study investigates the bioengineering potential of the marine diatom Phaeodactylum tricornutum through the introduction of cannabis genes, specifically, tetraketide synthase (TKS), and olivetolic acid cyclase (OAC), for the production of the cannabinoid precursor, olivetolic acid (OA). P. tricornutum is a promising biotechnological platform due to its fast growth rate, amenability to genetic manipulation, and ability to produce valuable compounds. Through genetic engineering techniques, we successfully integrated the cannabis genes TKS and OAC into the diatom. P. tricornutum transconjugants expressing these genes showed the production of the recombinant TKS and OAC enzymes, detected via Western blot analysis, and the production of cannabinoids precursor (OA) detected using the HPLC/UV spectrum when compared to the wild-type strain. Quantitative analysis revealed significant olivetolic acid accumulation (0.6-2.6 mg/L), demonstrating the successful integration and functionality of the heterologous genes. Furthermore, the introduction of TKS and OAC genes led to the synthesis of novel molecules, potentially expanding the repertoire of bioactive compounds accessible through diatom-based biotechnology. This study demonstrates the successful bioengineering of P. tricornutum with cannabis genes, enabling the production of OA as a precursor for cannabinoid production and the synthesis of novel molecules with potential pharmaceutical applications.

Differences in neurometabolites and transcranial magnetic stimulation motor maps in children with attention-deficit/hyperactivity disorder.

BACKGROUND: Although much is known about cognitive dysfunction in attention-deficit/hyperactivity disorder (ADHD), few studies have examined the pathophysiology of disordered motor circuitry. We explored differences in neurometabolite levels and transcranial magnetic stimulation (TMS)-derived corticomotor representations among children with ADHD and typically developing children. METHODS: We used magnetic resonance spectroscopy (MRS) protocols to measure excitatory (glutamate + glutamine [Glx]) and inhibitory (γ-aminobutyric acid [GABA]) neurometabolite levels in the dominant primary motor cortex (M1) and the supplementary motor area (SMA) in children with ADHD and typically developing children. We used robotic neuronavigated TMS to measure corticospinal excitability and create corticomotor maps. RESULTS: We collected data from 26 medication-free children with ADHD (aged 7-16 years) and 25 typically developing children (11-16 years). Children with ADHD had lower M1 Glx (p = 0.044, d = 0.6); their mean resting motor threshold was lower (p = 0.029, d = 0.8); their map area was smaller (p = 0.044, d = 0.7); and their hotspot density was higher (p = 0.008, d = 0.9). M1 GABA levels were associated with motor map area (p = 0.036).Limitations: Some TMS data were lost because the threshold of some children exceeded 100% of the machine output. The relatively large MRS voxel required to obtain sufficient signal-to-noise ratio and reliably measure GABA levels encompassed tissue beyond the M1, making this measure less anatomically specific. CONCLUSION: The neurochemistry and neurophysiology of key nodes in the motor network may be altered in children with ADHD, and the differences appear to be related to each other. These findings suggest potentially novel neuropharmacological and neuromodulatory targets for ADHD.

Reliability Validity and Responsiveness of the Spinal Cord Independence Measure 4th Version in a Multicultural Setup.

OBJECTIVE: To examine the fourth version of the Spinal Cord Independence Measure for reliability and validity. DESIGN: Partly blinded comparison with the criterion standard Spinal Cord Independence Measure III, and between examiners and examinations. SETTING: A multicultural cohort from 19 spinal cord injury units in 11 countries. PARTICIPANTS: A total of 648 patients with spinal cord injury. INTERVENTION: Assessment with Spinal Cord Independence Measure (SCIM IV) and Spinal Cord Independence Measure (SCIM III) on admission to inpatient rehabilitation and before discharge. MAIN OUTCOME MEASURES: SCIM IV interrater reliability, internal consistency, correlation with and difference from SCIM III, and responsiveness. RESULTS: Total agreement between examiners was above 80% on most SCIM IV tasks. All Kappa coefficients were above 0.70 and statistically significant (P<.001). Pearson's coefficients of the correlation between the examiners were above 0.90, and intraclass correlation coefficients were above 0.90. Cronbach's alpha was above 0.96 for the entire SCIM IV, above 0.66 for the subscales, and usually decreased when an item was eliminated. Reliability values were lower for the subscale of respiration and sphincter management, and on admission than at discharge. SCIM IV and SCIM III mean values were very close, and the coefficients of Pearson correlation between them were 0.91-0.96 (P<.001). The responsiveness of SCIM IV was not significantly different from that of SCIM III in most of the comparisons. CONCLUSIONS: The validity, reliability, and responsiveness of SCIM IV, which was adjusted to assess specific patient conditions or situations that SCIM III does not address, and which includes more accurate definitions of certain scoring criteria, are very good and quite similar to those of SCIM III. SCIM IV can be used for clinical and research trials, including international multi-center studies, and its group scores can be compared with those of SCIM III.

Validity and responsiveness of the Standing and Walking Assessment Tool for sub-acute traumatic spinal cord injury.

STUDY DESIGN: This is a retrospective longitudinal study. OBJECTIVE: The Standing and Walking Assessment Tool (SWAT) combines stages of standing and walking recovery (SWAT stages) with established measures (Berg Balance Scale (BBS), 10-m walk test (10MWT), 6-min walk test (6MWT), and modified Timed Up-and-Go (mTUG)). We evaluated the SWAT's validity (known-groups and convergent) and responsiveness among inpatients with sub-acute, traumatic spinal cord injury (SCI). SETTING: Ten Canadian rehabilitation hospitals. METHODS: Upon admission, SWAT stage and core measures (BBS, 10MWT, 6MWT, and mTUG), International Standards for Neurological Classification of SCI sensory and motor scores, and Spinal Cord Independence Measure III (SCIM) were collected from 618 adults with SCI. Known-groups validity was evaluated by comparing SWAT stage distributions across American Spinal Injury Association Impairment Scale (AIS) classification. Convergent validity was evaluated by correlating SWAT stages with scores on other measures using Spearman's rho. The SWAT (stage and core measures) was re-administered at discharge. To evaluate responsiveness, SWAT stages at admission and discharge were compared. The standardized response mean (SRM) was used to evaluate the responsiveness of core SWAT measures. RESULTS: The SWAT stage distribution of participants with AIS D injuries differed from those of participants with AIS A-C injuries (p ≤ 0.002). SWAT stages correlated strongly with BBS and motor scores (ρ = 0.778-0.836), and moderately with SCIM, mTUG, 10MWT, 6MWT, and sensory scores (ρ = 0.409-0.692). Discharge SWAT stage was greater than the admission stage (p < 0.0001). The BBS was the most responsive core SWAT measure (SRM = 1.26). CONCLUSIONS: The SWAT is a valid and responsive approach to the measurement of standing and walking ability during sub-acute SCI.

Cross-validation of ELISA and a portable surface plasmon resonance instrument for IgG antibody serology with SARS-CoV-2 positive individuals.

We report on the development of surface plasmon resonance (SPR) sensors and matching ELISAs for the detection of nucleocapsid and spike antibodies specific against the novel coronavirus 2019 (SARS-CoV-2) in human serum, plasma and dried blood spots (DBS). When exposed to SARS-CoV-2 or a vaccine against SARS-CoV-2, the immune system responds by expressing antibodies at levels that can be detected and monitored to identify the fraction of the population potentially immunized against SARS-CoV-2 and support efforts to deploy a vaccine strategically. A SPR sensor coated with a peptide monolayer and functionalized with various sources of SARS-CoV-2 recombinant proteins expressed in different cell lines detected human anti-SARS-CoV-2 IgG antibodies in clinical samples. Nucleocapsid expressed in different cell lines did not significantly change the sensitivity of the assays, whereas the use of a CHO cell line to express spike ectodomain led to excellent performance. This bioassay was performed on a portable SPR instrument capable of measuring 4 biological samples within 30 minutes of sample/sensor contact and the chip could be regenerated at least 9 times. Multi-site validation was then performed with in-house and commercial ELISA, which revealed excellent cross-correlations with Pearson's coefficients exceeding 0.85 in all cases, for measurements in DBS and plasma. This strategy paves the way to point-of-care and rapid testing for antibodies in the context of viral infection and vaccine efficacy monitoring.

Using wearable sensors to characterize gait after spinal cord injury: evaluation of test-retest reliability and construct validity.

STUDY DESIGN: Quantitative cross-sectional study. OBJECTIVES: Evaluate the test-retest reliability and the construct validity of inertial measurement units (IMU) to characterize spatiotemporal gait parameters in individuals with SCI. SETTING: Two SCI rehabilitation centers in Canada. METHODS: Eighteen individuals with SCI participated in two evaluation sessions spaced 2 weeks apart. Fifteen able-bodied individuals were also recruited. Participants walked 20 m overground under five conditions that challenged balance to varying degrees. Five IMU were attached to the lower-extremities and the sacrum to collect the mean and the coefficient of variation of five gait parameters (gait cycle time, double-support percentage, cadence, stride length, stride velocity). Intra-class correlation coefficients (ICC) were used to evaluate the test-retest reliability. Linear mixed-effects models were used to compare the five walking conditions to evaluate known-group validity while Spearman's correlation coefficients were used to characterize the level of association between gait parameters and the Mini BESTest (MBT). RESULTS: Cadence was reliable across all walking conditions. Reliability was higher for the mean (ICC = 0.55-0.98) of the parameters compared to their coefficient of variation (ICC = 0.16-0.97). Cadence collected with IMU had construct validity as their values differed across walking conditions and groups of participants. The coefficient of variation was generally better than the mean to show differences across the five walking conditions. The MBT was moderately to strongly associated with mean cadence (ρ ≥ 0.498) and its coefficient of variation (ρ ≤ -0.49) during most walking conditions. CONCLUSIONS: IMU provide reliable and valid measurements of gait parameters in ambulatory individuals with SCI.

Parental evaluation of a revised autism spectrum disorder diagnostic process for children under 36 months of age.

BACKGROUND: The Alberta Children's Hospital-Autism Spectrum Disorder Diagnostic Clinic (ACH-ASDC) was restructured due to long wait times and unsustainable clinic workflow. Major changes included the initiation of pre- and post-ASD parent education sessions and distinct ASD screening appointments before the ASD diagnostic appointment. METHODS: We conducted a parental program evaluation in summer 2018 of the ACH-ASDC. We used a cross-sectional survey to evaluate key outcomes including parental satisfaction, and the percentage of families obtaining access to government supports and early intervention programs. RESULTS: For the 101 eligible patients diagnosed with ASD under 36 months of age 70 (69.3%) parents agreed to participate. The mean diagnostic age of the children diagnosed with ASD was 30.6 months (SD=4.1 months). There were no statistically significant age differences between biological sexes. Ninety-three per cent of parents felt that ASD educational sessions were useful, and 92% of parents were satisfied to very satisfied with the overall ASD diagnostic process. Ninety per cent of parents had access to at least one of the key resources available for ASD early intervention in our province following diagnosis. Parents reported a positive impact on intervention provided to their child in the areas of communication, social interaction, and behaviour. CONCLUSION: Parents of children diagnosed with ASD expressed a high level of satisfaction with the restructured ACH-ASDC process. Implementing parent education sessions was well received and met parents' needs. Parents were able to access intervention services following diagnosis and reported positive impacts for their child. Re-envisioning program approaches to incorporate novel strategies to support families should be encouraged.

Impact of COVID-19 on lifestyle habits and mental health symptoms in children with attention-deficit/hyperactivity disorder in Canada.

OBJECTIVES: The COVID-19 pandemic created an environment of restricted access to health and recreation services. Lifestyle habits including sleep, eating, exercise, and screen use were modified, potentially exacerbating adverse mental health outcomes. This study investigates the impact of COVID-19 on lifestyle habits and mental health symptoms in paediatric attention-deficit/hyperactivity disorder (ADHD) in Canada. METHODS: An online survey was distributed across Canada to caregivers of children with ADHD (children aged 5 to 18 years) assessing depression (PHQ-9), anxiety (GAD-7), ADHD (SNAP-IV), and lifestyle behaviours. Data were analyzed by gender (male/female) and age category (5 to 8, 9 to 12, and 13 to 18 years). Spearman's correlations between lifestyle habits and mental health outcomes were conducted. RESULTS: A total of 587 surveys were completed. Mean child age was 10.14 years (SD 3.06), including 166 females (28.3%). The PHQ-9 and GAD-7 indicated that 17.4% and 14.1% of children met criteria for moderately severe to severe depression and anxiety symptoms respectively. Children met SNAP-IV cut-off scores for inattention (73.7%), hyperactivity/impulsivity (66.8%), and oppositional defiant disorder (38.6%) behaviours. Caregivers reported changes in sleep (77.5%), eating (58.9%), exercise (83.7%), and screen use (92.9%) in their ADHD child, greatly impacting youth. Sleeping fewer hours/night, eating more processed foods, and watching TV/playing videogames >3.5 hours/day correlated with greater depression, anxiety and ADHD symptoms, and exercising <1 hour/day further correlated with depression symptoms (P<0.01). CONCLUSIONS: The COVID-19 pandemic has resulted in less healthy lifestyle habits and increased mental health symptoms in Canadian children with ADHD. Longitudinal studies to better understand the relationship between these factors are recommended.

Comparative analysis of alternative polyadenylation in S. cerevisiae and S. pombe.

Alternative polyadenylation (APA) is a widespread mechanism that generates mRNA isoforms with distinct properties. Here we have systematically mapped and compared cleavage and polyadenylation sites (PASs) in two yeast species, S. cerevisiae and S. pombe Although >80% of the mRNA genes in each species were found to display APA, S. pombe showed greater 3' UTR size differences among APA isoforms than did S. cerevisiae PASs in different locations of gene are surrounded with distinct sequences in both species and are often associated with motifs involved in the Nrd1-Nab3-Sen1 termination pathway. In S. pombe, strong motifs surrounding distal PASs lead to higher abundances of long 3' UTR isoforms than short ones, a feature that is opposite in S. cerevisiae Differences in PAS placement between convergent genes lead to starkly different antisense transcript landscapes between budding and fission yeasts. In both species, short 3' UTR isoforms are more likely to be expressed when cells are growing in nutrient-rich media, although different gene groups are affected in each species. Significantly, 3' UTR shortening in S. pombe coordinates with up-regulation of expression for genes involved in translation during cell proliferation. Using S. pombe strains deficient for Pcf11 or Pab2, we show that reduced expression of 3'-end processing factors lengthens 3' UTR, with Pcf11 having a more potent effect than Pab2. Taken together, our data indicate that APA mechanisms in S. pombe and S. cerevisiae are largely different: S. pombe has many of the APA features of higher species, and Pab2 in S. pombe has a different role in APA regulation than its mammalian homolog, PABPN1.

Long noncoding RNA repertoire and targeting by nuclear exosome, cytoplasmic exonuclease, and RNAi in fission yeast.

Long noncoding RNAs (lncRNAs), which are longer than 200 nucleotides but often unstable, contribute a substantial and diverse portion to pervasive noncoding transcriptomes. Most lncRNAs are poorly annotated and understood, although several play important roles in gene regulation and diseases. Here we systematically uncover and analyze lncRNAs in Schizosaccharomyces pombe. Based on RNA-seq data from twelve RNA-processing mutants and nine physiological conditions, we identify 5775 novel lncRNAs, nearly 4× the previously annotated lncRNAs. The expression of most lncRNAs becomes strongly induced under the genetic and physiological perturbations, most notably during late meiosis. Most lncRNAs are cryptic and suppressed by three RNA-processing pathways: the nuclear exosome, cytoplasmic exonuclease, and RNAi. Double-mutant analyses reveal substantial coordination and redundancy among these pathways. We classify lncRNAs by their dominant pathway into cryptic unstable transcripts (CUTs), Xrn1-sensitive unstable transcripts (XUTs), and Dicer-sensitive unstable transcripts (DUTs). XUTs and DUTs are enriched for antisense lncRNAs, while CUTs are often bidirectional and actively translated. The cytoplasmic exonuclease, along with RNAi, dampens the expression of thousands of lncRNAs and mRNAs that become induced during meiosis. Antisense lncRNA expression mostly negatively correlates with sense mRNA expression in the physiological, but not the genetic conditions. Intergenic and bidirectional lncRNAs emerge from nucleosome-depleted regions, upstream of positioned nucleosomes. Our results highlight both similarities and differences to lncRNA regulation in budding yeast. This broad survey of the lncRNA repertoire and characteristics in S. pombe, and the interwoven regulatory pathways that target lncRNAs, provides a rich framework for their further functional analyses.

Postural control strategy after incomplete spinal cord injury: effect of sensory inputs on trunk-leg movement coordination.

BACKGROUND: Postural control is affected after incomplete spinal cord injury (iSCI) due to sensory and motor impairments. Any alteration in the availability of sensory information can challenge postural stability in this population and may lead to a variety of adaptive movement coordination patterns. Hence, identifying the underlying impairments and changes to movement coordination patterns is necessary for effective rehabilitation post-iSCI. This study aims to compare the postural control strategy between iSCI and able-bodied populations by quantifying the trunk-leg movement coordination under conditions that affects sensory information. METHODS: 13 individuals with iSCI and 14 aged-matched able-bodied individuals performed quiet standing on hard and foam surfaces with eyes open and closed. We used mean Magnitude-Squared Coherence between trunk-leg accelerations measured by accelerometers placed over the sacrum and tibia. RESULTS: We observed a similar ankle strategy at lower frequencies (f ≤ 1.0 Hz) between populations. However, we observed a decreased ability post-iSCI in adapting inter-segment coordination changing from ankle strategy to ankle-hip strategy at higher frequencies (f > 1.0 Hz). Moreover, utilizing the ankle-hip strategy at higher frequencies was challenged when somatosensory input was distorted, whereas depriving visual information did not affect balance strategy. CONCLUSION: Trunk-leg movement coordination assessment showed sensitivity, discriminatory ability, and excellent test-retest reliability to identify changes in balance control strategy post-iSCI and due to altered sensory inputs. Trunk-leg movement coordination assessment using wearable sensors can be used for objective outcome evaluation of rehabilitative interventions on postural control post-iSCI.

Widespread exon skipping triggers degradation by nuclear RNA surveillance in fission yeast.

Exon skipping is considered a principal mechanism by which eukaryotic cells expand their transcriptome and proteome repertoires, creating different splice variants with distinct cellular functions. Here we analyze RNA-seq data from 116 transcriptomes in fission yeast (Schizosaccharomyces pombe), covering multiple physiological conditions as well as transcriptional and RNA processing mutants. We applied brute-force algorithms to detect all possible exon-skipping events, which were widespread but rare compared to normal splicing events. Exon-skipping events increased in cells deficient for the nuclear exosome or the 5'-3' exonuclease Dhp1, and also at late stages of meiotic differentiation when nuclear-exosome transcripts decreased. The pervasive exon-skipping transcripts were stochastic, did not increase in specific physiological conditions, and were mostly present at less than one copy per cell, even in the absence of nuclear RNA surveillance and during late meiosis. These exon-skipping transcripts are therefore unlikely to be functional and may reflect splicing errors that are actively removed by nuclear RNA surveillance. The average splicing rate by exon skipping was ∼ 0.24% in wild type and ∼ 1.75% in nuclear exonuclease mutants. We also detected approximately 250 circular RNAs derived from single or multiple exons. These circular RNAs were rare and stochastic, although a few became stabilized during quiescence and in splicing mutants. Using an exhaustive search algorithm, we also uncovered thousands of previously unknown splice sites, indicating pervasive splicing; yet most of these splicing variants were cryptic and increased in nuclear degradation mutants. This study highlights widespread but low frequency alternative or aberrant splicing events that are targeted by nuclear RNA surveillance.

The nuclear poly(A)-binding protein interacts with the exosome to promote synthesis of noncoding small nucleolar RNAs.

Poly(A)-binding proteins (PABPs) are important to eukaryotic gene expression. In the nucleus, the PABP PABPN1 is thought to function in polyadenylation of pre-mRNAs. Deletion of fission yeast pab2, the homolog of mammalian PABPN1, results in transcripts with markedly longer poly(A) tails, but the nature of the hyperadenylated transcripts and the mechanism that leads to RNA hyperadenylation remain unclear. Here we report that Pab2 functions in the synthesis of noncoding RNAs, contrary to the notion that PABPs function exclusively on protein-coding mRNAs. Accordingly, the absence of Pab2 leads to the accumulation of polyadenylated small nucleolar RNAs (snoRNAs). Our findings suggest that Pab2 promotes poly(A) tail trimming from pre-snoRNAs by recruiting the nuclear exosome. This work unveils a function for the nuclear PABP in snoRNA synthesis and provides insights into exosome recruitment to polyadenylated RNAs.

Redesign of the autism spectrum screening and diagnostic process for children aged 12 to 36 months.

Diagnosing autism spectrum disorder (ASD) is challenging, resource-intense and time-consuming due to clinical and etiologic heterogeneity. With the rapid increase in prevalence of ASD, higher demand for diagnostic assessment often means long waitlists for families, and limited access to specialized intervention and support. In 2013, the Alberta Children's Hospital-Autism Spectrum Disorder Diagnostic Clinic (ACH-ASDC) experienced a significant waitlist in the 12 to 36 months' population. A Quality Improvement Project was started in 2014; one program aim was to create an efficient, sustainable and evidence-based ASD diagnostic evaluation process. The redesigned diagnostic process included: 1) pre- and postassessment parent information sessions, 2) a screening appointment and 3) standardized clinical appointment pathways. Within its first year, the new process reduced wait times to under a month without an increase in resources, leading to an efficient diagnostic process being sustained since its implementation.

Justify Your Answer: The Role of Written Think Aloud in Script Concordance Testing.

Construct: Clinical reasoning assessment is a growing area of interest in the medical education literature. Script concordance testing (SCT) evaluates clinical reasoning in conditions of uncertainty and has emerged as an innovative tool in the domain of clinical reasoning assessment. SCT quantifies the degree of concordance between a learner and an experienced clinician and attempts to capture the breadth of responses of expert clinicians, acknowledging the significant yet acceptable variation in practice under situations of uncertainty. BACKGROUND: SCT has been shown to be a valid and reliable clinical reasoning assessment tool. However, as SCT provides only quantitative information, it may not provide a complete assessment of clinical reasoning. APPROACH: Think aloud (TA) is a qualitative research tool used in clinical reasoning assessment in which learners verbalize their thought process around an assigned task. This study explores the use of TA, in the form of written reflection, in SCT to assess resident clinical reasoning, hypothesizing that the information obtained from the written TA would enrich the quantitative data obtained through SCT. Ninety-one pediatric postgraduate trainees and 21 pediatricians from 4 Canadian training centers completed an online test consisting of 24 SCT cases immediately followed by retrospective written TA. Six of 24 cases were selected to gather TA data. These cases were chosen to allow all phases of clinical decision making (diagnosis, investigation, and treatment) to be represented in the TA data. Inductive thematic analysis was employed when systematically reviewing TA responses. RESULTS: Three main benefits of adding written TA to SCT were identified: (a) uncovering instances of incorrect clinical reasoning despite a correct SCT response, (b) revealing sound clinical reasoning in the context of a suboptimal SCT response, and (c) detecting question misinterpretation. CONCLUSIONS: Written TA can optimize SCT by demonstrating when correct examinee responses are based on guessing or uncertainty rather than robust clinical rationale. TA can also enhance SCT by allowing examinees to provide justification for responses that otherwise would have been considered incorrect and by identifying questions that are frequently misinterpreted to avoid including them in future examinations. TA also has significant value in differentiating between acceptable variations in expert clinician responses and deviance associated with faulty rationale or question misinterpretation; this could improve SCT reliability. A written TA protocol appears to be a valuable tool to assess trainees' clinical reasoning and can strengthen the quantitative assessment provided by SCT.

Evolutions in clinical reasoning assessment: The Evolving Script Concordance Test.

INTRODUCTION: Script concordance testing (SCT) is a method of assessment of clinical reasoning. We developed a new type of SCT case design, the evolving SCT (E-SCT), whereby the patient's clinical story is "evolving" and with thoughtful integration of new information at each stage, decisions related to clinical decision-making become increasingly clear. OBJECTIVES: We aimed to: (1) determine whether an E-SCT could differentiate clinical reasoning ability among junior residents (JR), senior residents (SR), and pediatricians, (2) evaluate the reliability of an E-SCT, and (3) obtain qualitative feedback from participants to help inform the potential acceptability of the E-SCT. METHODS: A 12-case E-SCT, embedded within a 24-case pediatric SCT (PaedSCT), was administered to 91 pediatric residents (JR: n = 50; SR: n = 41). A total of 21 pediatricians served on the panel of experts (POE). A one-way analysis of variance (ANOVA) was conducted across the levels of experience. Participants' feedback on the E-SCT was obtained with a post-test survey and analyzed using two methods: percentage preference and thematic analysis. RESULTS: Statistical differences existed across levels of training: F = 19.31 (df = 2); p < 0.001. The POE scored higher than SR (mean difference = 10.34; p < 0.001) and JR (mean difference = 16.00; p < 0.001). SR scored higher than JR (mean difference = 5.66; p < 0.001). Reliability (Cronbach's α) was 0.83. Participants found the E-SCT engaging, easy to follow and true to the daily clinical decision-making process. CONCLUSIONS: The E-SCT demonstrated very good reliability and was effective in distinguishing clinical reasoning ability across three levels of experience. Participants found the E-SCT engaging and representative of real-life clinical reasoning and decision-making processes. We suggest that further refinement and utilization of the evolving style case will enhance SCT as a robust, engaging, and relevant method for the assessment of clinical reasoning.

Contribution of SWEET to improve paediatric diabetes care in developing countries.

Diabetes affects many children living in developing countries. Through an informal survey, five SWEET (Better control in Pediatric and Adolescent diabeteS: Working to crEate CEnTers of Reference) centers from developing countries (Mali, Costa Rica, Argentina and two from India) share their perspective on caring for children with diabetes. Each center provides a description of the population of children with diabetes they serve, the organization of care, and the challenges encountered on a daily basis in the provision of this care. In the second part, we summarize the anticipated benefits and challenges associated with participation in SWEET. This resulting article is a testimony of the reality of managing diabetes by dynamic teams striving to achieve recommended standards of care for pediatric diabetes in an environment with limited resources.