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ABSTRACT Tuberous sclerosis complex (TSC) is a variable multisystem disorder. The “no mutations identified” (NMI) group are reportedly phenotypically milder than those with an identified molecular cause, and often have mosaic or intronic variants not detected by standard sequencing methods. Methods We describe the phenotypes in an Australian TSC NMI group (n = 18) and a molecular testing strategy implementable in a diagnostic laboratory. Massively parallel sequencing (MPS) of the whole genomic regions of TSC1 and TSC2 was performed using DNA extracted from multiple tissue samples per participant. Results Our study showed that the phenotype in TSC NMI individuals can be similar to those with heterozygous, particularly TSC1, variants. Although neurodevelopmental outcomes can be less severe, the number of organ systems involved was similar to the non‐mosaic groups. A diagnostic yield of 72% (13/18) was achieved, with the majority (10/13) being mosaic variants and the remainder heterozygous variants missed on previous testing. Conclusion Testing DNA from multiple tissue samples allowed for validation of otherwise discarded low‐level mosaic variants and detection of mosaic variants by MPS without excessive cost or the need for specialised techniques. Implementing this approach in a diagnostic setting is viable and allows optimal clinical care of patients with NMI TSC.

Abstract Episomal AMA1‐based plasmids are increasingly used for expressing biosynthetic pathways and CRISPR/Cas systems in filamentous fungi cell factories due to their high transformation efficiency and multicopy nature. However, the gene expression from AMA1 plasmids has been observed to be highly heterogeneous in growing mycelia. To overcome this limitation, here we developed next‐generation AMA1‐based plasmids that ensure homogeneous and strong expression. We achieved this by evaluating various degradation tags fused to the auxotrophic marker gene on the AMA1 plasmid, which introduces a more stringent selection pressure throughout multicellular fungal growth. With these improved plasmids, we observed in Aspergillus nidulans a 5‐fold increase in the expression of a fluorescent reporter, a doubling in the efficiency of a CRISPRa system for genome mining, and a up to a 10‐fold increase in the production of heterologous natural product metabolites. This strategy has the potential to be applied to diverse filamentous fungi.

Abstract Background This study sought to identify cognitive and behavioral predictors of COVID‐19 vaccine hesitancy. Specifically, this study examined the effect of anxiety about developing COVID‐19 and proactive coping behavior on the likelihood of reporting COVID‐19 vaccine hesitancy in a sample of adults living in the United States. Methods An online survey of proactive coping strategies, anxiety related to developing COVID‐19, and vaccine hesitancy was administered in October 2020 to 534 adults aged 21–79‐years old. Age, gender, race, self‐rated health, years of education, COVID‐19 knowledge, and perceived constraints were included as covariates. Results Over half of the study participants (56.7%) were COVID‐19 vaccine hesitant. People who were less anxious about developing COVID‐19 were more likely to be vaccine hesitant. A statistically significant COVID‐19 anxiety × proactive coping interaction showed the odds of vaccine hesitancy was highest among individuals with low anxiety about developing COVID‐19 and high proactive coping, whereas vaccine hesitancy was lowest among individuals with high COVID‐19 anxiety and high proactive coping. Conclusion Results support a future‐oriented approach to public health outreach efforts regarding COVID‐19 vaccines. Improvement of proactive coping skills and emphasis on the likelihood of contracting COVID‐19 may be more effective in increasing vaccine uptake than simply restating scientific facts regarding safety or efficacy.

Abstract Smith‐Lemli‐Opitz syndrome (SLOS) is a relatively common genetic cause of developmental delay and may only present in conjunction with 2,3 toe syndactyly. This case series illustrates a milder phenotype of SLOS, where the predominant findings are neurocognitive in the presence of 2,3 toe syndactyly.

Abstract Background and Aim In 2017, ustekinumab (UST) was included in Israel's National Basket of Health Services for treatment of biologic‐experienced Crohn's disease (CD) patients with moderately to severely active disease. This study aims to provide real‐world evidence on persistence and clinical outcomes among early users of UST. Methods This retrospective cohort study was conducted using data from Maccabi Healthcare Services (MHS; 2.5‐million‐member state‐mandated health provider, Israel). Adult patients with a CD diagnosis code who had ≥1 dispensed UST prescription in 2017–2018 and at least 12 months of prior continuous health plan enrollment were included. Outcomes, including treatment discontinuation, dose‐escalation (based on shortened intervals between purchases), CD‐related surgery, CD‐related hospitalization, corticosteroid (CS) discontinuation, and use of opioids were evaluated from the date of first dispensed UST through the end of 2019 using Kaplan–Meier analysis. Results A total of 162 eligible patients (81 [49.4%] female; median age 34.4 years [IQR 23.2–46.3]; median years since CD diagnosis 8.6 [IQR 4·8–16.0]) were enrolled in the study. Discontinuation rate after 365 and 540 days of follow‐up was 27.8% and 35.6%. Dose escalation was estimated at 15.4% and 28.6%, respectively. The first‐year cumulative rate of CD‐related surgery and CD‐related hospitalization were estimated at 4.7% and 9.8%, respectively. Conclusion In this real‐world CD cohort of UST users, results suggest persistence is relatively high as compared to other biologics for CD. Comparative effectiveness of different biologic treatments for CD in this population should be further explored.