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Summary: Blood lipid traits are treatable and heritable risk factors for heart disease, a leading cause of mortality worldwide. Although genome-wide association studies (GWASs) have discovered hundreds of variants associated with lipids in humans, most of the causal mechanisms of lipids remain unknown. To better understand the biological processes underlying lipid metabolism, we investigated the associations of plasma protein levels with total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL) cholesterol, and low-density lipoprotein (LDL) cholesterol in blood. We trained protein prediction models based on samples in the Multi-Ethnic Study of Atherosclerosis (MESA) and applied them to conduct proteome-wide association studies (PWASs) for lipids using the Global Lipids Genetics Consortium (GLGC) data. Of the 749 proteins tested, 42 were significantly associated with at least one lipid trait. Furthermore, we performed transcriptome-wide association studies (TWASs) for lipids using 9,714 gene expression prediction models trained on samples from peripheral blood mononuclear cells (PBMCs) in MESA and 49 tissues in the Genotype-Tissue Expression (GTEx) project. We found that although PWASs and TWASs can show different directions of associations in an individual gene, 40 out of 49 tissues showed a positive correlation between PWAS and TWAS signed p values across all the genes, which suggests high-level consistency between proteome-lipid associations and transcriptome-lipid associations.

Adolescence is a period of profound biopsychosocial development, with pubertally-driven neural reorganization as social demands increase in peer contexts. The explosive increase in social media access has fundamentally changed peer interactions among youth, creating an urgent need to understand its impact on neurobiological development and mental health. Extant literature indicates that using social media promotes social comparison and feedback seeking (SCFS) behaviors in youth, which portend increased risk for mental health disorders, but little is known about its impact on neurobiological development. We assessed social media behaviors, mental health symptoms, and spontaneous cortical activity using magnetoencephalography (MEG) in 80 typically developing youth (8–16 years) and tested how self-reported pubertal stage moderates their relationship. More mature adolescents who engaged in more SCFS showed weaker fusiform/parahippocampal alpha and medial prefrontal beta activity, and increased symptoms of anxiety and attention problems. Engaging in SCFS on social media during adolescence may thus relate to developmental differences in brain regions that undergo considerable development during puberty. These results are consistent with works indicating altered neurodevelopmental trajectories within association cortices surrounding the onset of many mental health disorders. Importantly, later pubertal stages may be most sensitive to the detrimental effects of social media use.

Background: Digital pathology systems (DPS) are emerging as capable technologies for clinical practice. Studies have analyzed pathologists' diagnostic concordance by comparing reviews of whole slide images (WSIs) to glass slides (e.g., accuracy). This observational study evaluated the reproducibility of pathologists' diagnostic reviews using the Aperio GT 450 DX under slightly different conditions (precision). Method: Diagnostic precision was tested in three conditions: intra-system (within systems), inter-system/site (between systems/sites), and intra- and inter-pathologist (within and between pathologists). A total of five study/reading pathologists (one pathologist each for intra-system, inter-system/site, and three for intra-pathologist/inter-pathologist analyses) were assigned to the respective sub-studies.A panel of 69 glass slides with 23 unique histological features was used to evaluate the WSI system's precision. Each glass slide was scanned to generate a unique WSI. From each WSI, the field of view (FOV) was generated (at least 2 FOVs/WSI), which included the selected features (1–3 features/FOV). Each pathologist reviewed the digital slides and identified which morphological features, if any, were present in each defined FOV. To minimize recall bias, an additional 12 wild card slides from different organ types were used for which FOVs were extracted. The pathologists also read these wild card slides FOVs; however, the corresponding feature identification was not included in the final data analysis. Results: Each measured endpoint met the pre-defined acceptance criteria of the lower bound of the 95% confidence interval (CI) overall agreement (OA) rate being ≥85% for each sub-study. The lower bound of the 95% CI for the intra-system OA rate was 95.8%; for inter-system analysis, it was 94.9%; for intra-pathologist analysis, 92.4%, whereas for inter-pathologist analyses, the lower bound of the 95% CI of the OA was 90.6%. Conclusion: The study results indicate that pathologists using the Aperio GT 450 DX WSI system can precisely identify histological features that may be required for accurately diagnosing anatomic pathology cases.

The vector genome (vg) titer measurement, which is used to control patient dosing and ensure control over drug product manufacturing, is essential for the development of recombinant adeno-associated virus (AAV) gene therapy products. While qPCR and droplet digital PCR technologies are commonly implemented for measuring vg titer, chromatographic techniques with UV detectors represent promising future approaches, in line with traditional biotherapeutics. Here, we introduce a novel vg titer measurement approach using size-exclusion high-performance liquid chromatography with UV detection, which achieves excellent method precision (

Objectives: The association between SARS-CoV-2 spike protein and cerebrovascular diseases raised a concern of cerebrovascular safety of COVID-19 vaccines. We aimed to determine the risk of radiologic cerebral small vessel disease (cSVD) progression with BNT162b2 and CoronaVac. Methods: In this community-based prospective cohort study, community-dwelling subjects underwent brain magnetic resonance imaging (MRI) before and 4 months after vaccination with BNT162b2 or CoronaVac. Unvaccinated subjects received serial brain MRI over a comparable interval. The primary outcome was progression of a composite of six standard cSVD biomarkers. We compared the risk of cSVD progression between vaccinated and unvaccinated subjects and identified predictors of primary outcome within each vaccine subgroup. Results: Of the 415 subjects recruited, 190 received BNT162b2, 152 received CoronaVac, and 73 remained unvaccinated. A total of 60 (14.4%) had COVID-19 infection before follow-up MRI, and 109 (26.3%) developed the primary outcome. Neither BNT162b2 (adjusted odds ratio [aOR] 0.61, 95% confidence interval [CI] 0.30-1.26, P = 0.179) nor CoronaVac (aOR 0.71, 95% CI 0.34-1.47, P = 0.349) was associated with cSVD progression. Among the BNT162b2 recipients, a higher surrogate virus neutralization test was associated (aOR 0.97, 95% CI 0.95-0.99, P = 0.002) with a lower risk of cSVD progression. Conclusions: BNT162b2 and CoronaVac did not increase cSVD burden in community-dwelling citizens. The association between surrogate virus neutralization test and cSVD progression among BNT162b2 recipients requires further investigation.

Adeno-associated virus (AAV) gene therapy vectors, which contain a DNA transgene packaged into a protein capsid, have shown tremendous therapeutic potential in recent years. Methods traditionally used in quality control labs, such as high-performance liquid chromatography (HPLC) and capillary electrophoresis (CE), do not provide a complete understanding of capsid viral protein (VP) charge heterogeneity. In the present study, we developed simple, one-step sample preparation and charge-based VP separation using imaged capillary isoelectric focusing (icIEF) for monitoring AAV products. The robustness of the method was confirmed through a design of experiments (DoE) exercise. An orthogonal reverse-phase (RP) HPLC method coupled with mass spectrometry was developed to separate and identify charge species. Additionally, capsid point mutants demonstrate the capability of the method to resolve deamidation at a single site on the viral proteins. Finally, case studies using two different AAV serotype vectors establish the icIEF method as stability indicating and demonstrate that increases in acidic species measured by icIEF correlate with increased deamidation, which, we show, results in decreased transduction efficiency. The addition of a rapid and robust icIEF method to the AAV capsid analytical toolkit enables development and consistent manufacturing of well-characterized gene therapy products.

Background: Acromial morphology is an important pathophysiological factor for the development of subacromial impingement syndrome. There are 3 radiological methods to evaluate acromial morphology: Bigliani, modified Epstein, and acromial angle. However, their reliability have not been compared in a single study, nor using standardized radiographs. Consequently, the evaluation of acromial morphology is currently not validated though its widespread use across the world. The objective of this study was to investigate reliability of the 3 known classifications and the novel Copenhagen Acromial Curve classification. Methods: Three experienced clinicians rated 102 standardized supraspinatus outlet view radiographs with the 4 classification methods in 2 separate sessions a month apart. All measurements were blinded. With an expected kappa (κ) and intraclass correlation coefficient (ICC) > 0.7 (+/−0.15), the target sample size was 87 radiographs. Results: The Bigliani classification had interrater and intrarater reliability ranging from fair to good (κ 0.32-0.41 and 0.26-0.62). The modified Epstein classification had fair to good interrater and intrarater reliability (κ 0.24-0.69 and 0.57-0.63). The acromial angle classification had moderate to good interrater and intrarater reliability (κ 0.53-0.60 and 0.59-0.72). The novel Copenhagen Acromial Curve classification showed moderate to good interrater and intrarater reliability (ICC 0.66-0.71 and 0.75-0.78, respectively). Conclusion: The Copenhagen Acromial Curve was the only classification method with an ICC value > 0.7. The popular Bigliani classification had the worst reliability. The Copenhagen Acromial Curve classification produces numerical data, as opposed to the other 3 classification methods. This could potentially be utilized in future research to establishing cut-off values for treatment stratification.

Background: Current protein biomarkers are only moderately predictive at identifying individuals with mild traumatic brain injury or concussion. Therefore, more accurate diagnostic markers are needed for sport-related concussion. Methods: This was a multicenter, prospective, case-control study of athletes who provided blood samples and were diagnosed with a concussion or were a matched non-concussed control within the National Collegiate Athletic Association–Department of Defense Concussion Assessment, Research, and Education Consortium conducted between 2015 and 2019. The blood was collected within 48 h of injury to identify protein abnormalities at the acute and subacute timepoints. Athletes with concussion were divided into 6 h post-injury (0–6 h post-injury) and after 6 h post-injury (7–48 h post-injury) groups. We applied a highly multiplexed proteomic technique that used a DNA aptamers assay to target 1305 proteins in plasma samples from athletes with and without sport-related concussion. Results: A total of 140 athletes with concussion (79.3% males; aged 18.71 ± 1.10 years, mean ± SD) and 21 non-concussed athletes (76.2% males; 19.14 ± 1.10 years) were included in this study. We identified 338 plasma proteins that significantly differed in abundance (319 upregulated and 19 downregulated) in concussed athletes compared to non-concussed athletes. The top 20 most differentially abundant proteins discriminated concussed athletes from non-concussed athletes with an area under the curve (AUC) of 0.954 (95% confidence interval: 0.922‒0.986). Specifically, after 6 h of injury, the individual AUC of plasma erythrocyte membrane protein band 4.1 (EPB41) and alpha-synuclein (SNCA) were 0.956 and 0.875, respectively. The combination of EPB41 and SNCA provided the best AUC (1.000), which suggests this combination of candidate plasma biomarkers is the best for diagnosing concussion in athletes after 6 h of injury. Conclusion: Our data suggest that proteomic profiling may provide novel diagnostic protein markers and that a combination of EPB41 and SNCA is the most predictive biomarker of concussion after 6 h of injury.

Background: Both anatomic total shoulder arthroplasty (aTSA) and reverse total shoulder arthroplasty (RTSA) are being increasingly performed. In the event of a complication necessitating revision, RTSA is more commonly performed in both scenarios. The purpose of this study was to compare clinical outcomes between patients undergoing revision RTSA for failed primary anatomic versus reverse total shoulder arthroplasty. Methods: We performed a retrospective review of a prospective single-institution shoulder arthroplasty database. All revision RTSAs performed between 2007 and 2019 with a minimum 2-year clinical follow-up were initially included. After excluding patients with a preoperative diagnosis of infection, an oncologic indication, or incomplete outcomes assessment, we included 45 revision RTSAs performed for failed primary aTSA and 15 for failed primary RTSA. Demographics, surgical characteristics, active range of motion (external rotation [ER], internal rotation, forward elevation [FE], abduction), outcome scores (American Shoulder and Elbow Surgeons score, Constant Score, Shoulder Pain and Disability Index, Simple Shoulder Test, and University of California, Los Angeles score), and the incidence of postoperative complications was compared between groups. Results: Primary aTSA was most often indicated for degenerative joint disease (82%), whereas primary RTSA was more often indicated for rotator cuff arthropathy (60%). On bivariate analysis, no statistically significant differences in any range of motion or clinical outcome measure were found between revision RTSA performed for failed aTSA vs. RTSA. On multivariate linear regression analysis, revision RTSA performed for failed aTSA vs. RTSA was not found to significantly influence any outcome measure. Humeral loosening as an indication for revision surgery was associated with more favorable outcomes for all four range of motion measures and all five outcome scores assessed. In contrast, an indication for revision of peri-prosthetic fracture was associated with poorer outcomes for three of four range of motion measures (ER, FE, abduction) and four of five outcome scores (Constant, Shoulder Pain and Disability Index, Simple Shoulder Test, University of California, Los Angeles). A preoperative diagnosis of fracture was associated with a poorer postoperative range of motion in ER, FE, and abduction, but was not found to significantly influence any outcome score. However, only two patients in our cohort had this indication. Complication and re-revision rates after revision RTSA for failed primary aTSA and RTSA were 27% and 9% vs. 20% and 14% (P = .487 and P = .515), respectively. Conclusion: Clinical outcomes of patients undergoing revision RTSA for failed primary shoulder arthroplasty did not significantly differ based on whether aTSA or RTSA was initially performed. However, larger studies are needed to definitively ascertain the influence of the primary construct on the outcomes of revision RTSA.

Cotton (Gossypium hirsutum L.) is an important fiber cash crop, but its root traits related to phosphorus (P) acquisition, including mycorrhizal root traits, are poorly understood. Eight cotton varieties bred in northwestern China that were released between 1950 and 2013 were grown in pots with or without one arbuscular mycorrhizal fungal (AMF) species (Funneliformis mosseae) at three P supply levels (0, 50 and 300 mg P as KH2PO4 kg−1). Eleven root traits were measured and calculated after 7 wk of growth. The more recent accessions had smaller root diameters, acquired less P and produced less biomass, indicating an (inadvertent) varietal selection for thinner roots that provided less cortical space for AMF, which then increased the need for a high P fertilizer level. At the two lower P levels, the mycorrhizal plants acquired more P and produced more biomass than non-mycorrhizal plants (3.2 vs. 0.9 mg P per plant; 1.8 vs. 0.9 g biomass per plant at P0; 14.5 vs. 1.7 mg P per plant; and 4.7 vs. 1.6 g biomass per plant at P50). At the highest P level, the mycorrhizal plants acquired more P than non-mycorrhizal plants (18.8 vs. 13.4 mg per P plant), but there was no difference in biomass (6.2 vs. 6.3 g per plant). At the intermediate P level, root diameter was significantly positively correlated with shoot biomass, P concentration and the P content of mycorrhizal plants. The results of our study support the importance of the outsourcing model of P acquisition in the root economics space framework. Inadvertent varietal selection in the last decades, resulting in thinner roots and a lower benefit from AMF, has led to a lower productivity of cotton varieties at moderate P supply (i.e., when mycorrhizal, the average biomass of older varieties 5.0 g per plant vs. biomass of newer varieties 4.4 g per plant), indicating the need to rethink cotton breeding efforts in order to achieve high yields without very high P input. One feasible way to solve the problem of inadvertent varietal selection for cotton is to be aware of the trade-offs between the root do-it-yourself strategy and the outsourcing towards AMF strategy, and to consider both morphological and mycorrhizal root traits when breeding cotton varieties.

Xanthogranuloma (XG) of the sellar region is a non-neoplastic inflammatory lesion characterized histologically by recent and remote hemorrhage, necrotic debris, fibrosis, chronic inflammation, and cholesterol clefts with associated foreign-body giant cells. The inflammatory lesion was recognized by the World Health Organization in 2000. XG of the sellar region is rare. Cases of pituitary adenoma (PA) with an associated XG (PA/XG) are extremely rare, with a total of 16 cases in the literature. PA/XG lacks specific clinical and radiologic signs, making pre-operative diagnosis challenging. Herein, we report a case of PA/XG, describe the radiologic and pathologic findings, and discuss the role of so-called silent or “subclinical pituitary apoplexy” in the possible histogenesis of PA/XGs.

Background: Poorer health outcomes experienced by young women with acute coronary syndrome may be related to sex differences in the safety and efficacy of antiplatelet agents, such as clopidogrel. Polymorphisms in drug metabolism enzyme (cytochrome P450 [CYP] family) genes are independent factors for the variability in response to clopidogrel. However, a sex-specific impact of genetics to explain worse clinical outcomes in women has not been explored extensively. Therefore, our objective was to determine whether an interaction of sex with CYP variants occurs among users of clopidogrel, and if so, its impact on 1-year adverse clinical outcomes. Methods: We used data from a combined cohort of 2272 patients (median age 49 years; 56% female) hospitalized for acute coronary syndrome. We examined interactions between sex and CYP variants among clopidogrel users at admission and discharge to assess associations with 1-year readmission due to cardiac events. Results: The case-only analysis of 177 participants on clopidogrel at the time of presentation showed that the risk of an atherothrombotic event was greater in female carriers of the CYP2C9∗3 loss-of-function allele (odds ratio = 3.77, 95% confidence interval = 1.54-9.24). The results of the multivariable logistic regression model for users of clopidogrel at discharge (n = 1733) indicated that women had significantly higher risk of atherothrombotic readmissions at 1 year (odds ratio = 1.55, 95% confidence interval = 1.16-2.07), compared to the risk for men, but the loss-of-function alleles, either individually or through a genetic risk score, were not associated with 1-year readmissions. Conclusion: This study highlights the need for an improved understanding of the role of sex-by-gene interactions in causing sex differences in drug metabolism. Résumé: Contexte: Les piètres résultats cliniques observés chez les jeunes femmes atteintes d’un syndrome coronarien aigu pourraient être liés à des différences entre les sexes en ce qui concerne l’innocuité et l’efficacité des antiplaquettaires, comme le clopidogrel. Le polymorphisme génétique des enzymes intervenant dans le métabolisme des médicaments (famille des cytochromes P450 [CYP]) est un facteur indépendant de la variabilité de la réponse au clopidogrel. Cependant, jamais la possibilité d’un effet génétique propre au sexe qui expliquerait les résultats cliniques défavorables chez les femmes n’a été examinée en profondeur. Notre objectif était donc de déterminer s’il se produit une interaction entre le sexe et les variants de CYP chez les personnes prenant du clopidogrel et, si tel est le cas, quels sont ses effets sur les résultats cliniques indésirables après un an. Méthodologie: Nous avons utilisé les données d’une cohorte combinée de 2 272 patients (âge médian : 49 ans; 56 % de femmes) hospitalisés en raison d’un syndrome coronarien aigu. Nous avons examiné les interactions entre le sexe et les variants du CYP chez les utilisateurs de clopidogrel au moment de leur admission à l’hôpital et de leur congé pour évaluer les liens entre ces variables et les réhospitalisations après un an en raison d’événements cardiaques. Résultats: L’analyse de cas de 177 participants prenant du clopidogrel au moment de leur admission à l’hôpital a montré que le risque d’événement athérothrombotique était plus élevé chez les femmes porteuses de l’allèle non fonctionnel CYP2C9∗3 (rapport de cotes = 3,77; intervalle de confiance [IC] à 95 % = 1,54 à 9,24). Les résultats obtenus avec le modèle de régression logistique multivariée pour les utilisateurs de clopidogrel au moment du congé (n = 1 733) indiquent que les femmes présentaient un risque significativement plus élevé de réhospitalisation en raison d’un événement athérothrombotique après un an (rapport de cotes = 1,55; [IC] à 95 % = 1,16 à 2,07), comparativement aux hommes. Toutefois, les allèles non fonctionnels, considérés individuellement ou sur la base d’un score de risque génétique, n’étaient pas liés à la réhospitalisation après un an. Conclusion: Cette étude fait ressortir la nécessité de mieux comprendre le rôle de l’interaction sexe-gène dans les différences entre les sexes relativement au métabolisme des médicaments.

Purpose: The purpose of this study was to use a smartphone app to collect gait metrics in patients undergoing hip arthroscopy for femoroacetabular impingement syndrome (FAIS) and determine when gait metrics return to preoperative levels and if gait metrics correlate with postoperative hip-specific patient-reported outcomes. Methods: Patients undergoing primary hip arthroscopy for FAIS from August 2021 to December 2022 were prospectively enrolled. Patients downloaded the iPhone app, rHip, at ∼6 months postoperatively, enabling retroactive access to Apple Health data. Gait metrics included step count, step length, and walking speed. Each patient’s preoperative baseline was compared to that individual’s successive postoperative week metrics. All patients underwent a standard 4-stage rehabilitation protocol. We compared metrics from beginning to end of each phase (i.e., week 1 to 6 for phase 1) and evaluated correlations with patient outcomes. Results: Fifty patients (40 women; age: 31.9 ± 14.5 years) were included. Patients had significantly reduced step count from postoperative weeks 1 to 5 and improved upon baseline at weeks 13 and 16 to 26. Patients had significantly reduced step length from postoperative weeks 1 to 4 and improved upon baseline at weeks 16 and 20. Patients had significantly lower walking speed from postoperative weeks 1 to 6 and improved upon baseline at week 20 (P < .05). Step count significantly improved over phase 1 rehabilitation (P < .05). Step length and walking speed significantly improved over each of phases 1 to 3 (P < .05 for all). Preoperatively, weak correlations were noted between step length and Patient-Reported Outcomes Measurement Information System (PROMIS) for Physical Function (PF)/Pain, as well as walking speed and Hip Outcome Score–Activities of Daily Living (HOS-ADL)/PROMIS-Pain. At 3 months, moderate correlations were noted between step count and HOS-ADL/12-item international Hip Outcome Tool (iHOT-12), step length and HOS-ADL/PROMIS-PF, and walking speed and HOS-ADL/iHOT-12/PROMIS-PF/PROMIS-Pain. At 6 months, weak to moderate correlations were noted between all 3 gait metrics and HOS-ADL. Conclusions: A smartphone app was capable of capturing health data gait metrics. Patients undergoing hip arthroscopy for FAIS returned to baseline levels in step count, step length, and walking speed after phase 1 (6 weeks) of rehabilitation. The most consistent correlations between gait metrics and step count were seen at 3 months, although only weak to moderate. Gait metrics similarly had weak to moderate correlations with HOS-ADL at 6 months. Level of Evidence: Level IV, case series.

While it is well established that the KEAP1-NRF2 pathway regulates the main inducible cellular response to oxidative stress, this cytoprotective function of NRF2 could become deleterious to the host if it confers survival onto irreparably damaged cells. In this regard, we have found that in diseased states, NRF2 promotes the transcriptional activation of a specific subset of the senescence-associated secretory phenotype (SASP) gene program, which we have named the NRF2-induced secretory phenotype (NISP). In two models of hepatic disease using Pten::Keap1 and Keap1::Atg7 double knockout mice, we found that the NISP functions in the liver to recruit CCR2 expressing monocytes, which function as immune system effector cells to directly remove the damaged cells. Through activation of this immune surveillance pathway, in non-transformed cells, NRF2 functions as a tumour suppressor to mitigate the long-term survival of damaged cells which otherwise would be detrimental for host survival. This pathway represents the final stage of the oxidative stress response, as it allows cells to be safely removed if the macromolecular damage caused by the original stressor is so extensive that it is beyond the repair capacity of the cell.

Summary: Each human genome has tens of thousands of rare genetic variants; however, identifying impactful rare variants remains a major challenge. We demonstrate how use of personal multi-omics can enable identification of impactful rare variants by using the Multi-Ethnic Study of Atherosclerosis, which included several hundred individuals, with whole-genome sequencing, transcriptomes, methylomes, and proteomes collected across two time points, 10 years apart. We evaluated each multi-omics phenotype’s ability to separately and jointly inform functional rare variation. By combining expression and protein data, we observed rare stop variants 62 times and rare frameshift variants 216 times as frequently as controls, compared to 13–27 times as frequently for expression or protein effects alone. We extended a Bayesian hierarchical model, “Watershed,” to prioritize specific rare variants underlying multi-omics signals across the regulatory cascade. With this approach, we identified rare variants that exhibited large effect sizes on multiple complex traits including height, schizophrenia, and Alzheimer’s disease.

Surgical repair of proximal hamstring injuries can relieve pain and restore lower extremity function in active individuals. Whereas traditional surgical techniques are performed via an open approach, more recent endoscopic proximal hamstring repair techniques have proven safe, effective, and potentially associated with fewer complications than open repair. One theorized disadvantage of existing endoscopic techniques is reduced security at the suture-tendon interface, as compared to open surgery, during which a running suture technique, such as a Krackow stitch, may be employed. In this article, we present a technique for increasing suture purchase by performing an all-endoscopic, running, locking stitch during proximal hamstring repair.

Summary: The balance of contralateral and ipsilateral retinogeniculate projections is critical for binocular vision, but the transcriptional programs regulating this process remain ill defined. Here we show that the Pou class homeobox protein POU3F1 is expressed in nascent mouse contralateral retinal ganglion cells (cRGCs) but not ipsilateral RGCs (iRGCs). Upon Pou3f1 inactivation, the proportion of cRGCs is reduced in favor of iRGCs, leading to abnormal projection ratios at the optic chiasm. Conversely, misexpression of Pou3f1 in progenitors increases the production of cRGCs. Using CUT&RUN and RNA sequencing in gain- and loss-of-function assays, we demonstrate that POU3F1 regulates expression of several key members of the cRGC gene regulatory network. Finally, we report that POU3F1 is sufficient to induce RGC-like cell production, even in late-stage retinal progenitors of Atoh7 knockout mice. This work uncovers POU3F1 as a regulator of the cRGC transcriptional program, opening possibilities for optic nerve regenerative therapies.

Rib hyperostosis has previously been described in conjunction with disorders causing excessive vertebral ossification due to osseous bridging across the costovertebral joint, such as in diffuse idiopathic skeletal hyperostosis. Hyperostosis is believed to be a reactive process due to altered forces across the affected rib as bridging osteophytes decrease mobility at the respective costovertebral joint. The imaging characteristics of rib hyperostosis can be highly suspicious for malignancy. We share 2 cases of biopsy-proven benign rib hyperostosis with imaging across multiple modalities in hopes of increasing awareness of this entity and its imaging characteristics. In the first case, a 62-year-old female without history of malignancy underwent rib biopsy after bone scintigraphy demonstrated intense radiotracer uptake along a posteromedial rib. In the second case, a 66-year-old male with history of recurrent prostate cancer underwent rib biopsy after interval development of intense radiotracer uptake on bone scintigraphy along a posteromedial rib, new compared to 6 months prior. Both cases were seen in the setting of osseous bridging at the respective costovertebral joint. Imaging findings include contiguous radiotracer uptake on bone scintigraphy confined to the rib and respective costovertebral joint, cortical bone thickening with osseous excrescence at the costovertebral joint on radiographic and cross-sectional imaging, and increased osseous edema-like change, postcontrast enhancement, and surrounding soft tissue edema on magnetic resonance imaging. By increasing awareness to these imaging features, we hope to improve diagnostic confidence and decrease unnecessary, expensive, and sometimes invasive workup for future patients.

The contemporary treatment of hamstring avulsions has been evolving, as more patients are being identified as having persistently symptomatic partial hamstring tears recalcitrant to nonoperative treatment. The endoscopic hamstring repair allows surgeons improved visualization of the footprint, as well as safe dissection of the sciatic nerve. The present technique article provides a step-by-step technical note to allow for safe and effective surgical treatment of partial hamstring tears.

Summary: Despite substantial lignocellulose conversion during mycelial growth, previous transcriptome and proteome studies have not yet revealed how secretomes from the edible mushroom Agaricus bisporus develop and whether they modify lignin models in vitro. To clarify these aspects, A. bisporus secretomes collected throughout a 15-day industrial substrate production and from axenic lab-cultures were subjected to proteomics, and tested on polysaccharides and lignin models. Secretomes (day 6–15) comprised A. bisporus endo-acting and substituent-removing glycoside hydrolases, whereas β-xylosidase and glucosidase activities gradually decreased. Laccases appeared from day 6 onwards. From day 10 onwards, many oxidoreductases were found, with numerous multicopper oxidases (MCO), aryl alcohol oxidases (AAO), glyoxal oxidases (GLOX), a manganese peroxidase (MnP), and unspecific peroxygenases (UPO). Secretomes modified dimeric lignin models, thereby catalyzing syringylglycerol-β-guaiacyl ether (SBG) cleavage, guaiacylglycerol-β-guaiacyl ether (GBG) polymerization, and non-phenolic veratrylglycerol-β-guaiacyl ether (VBG) oxidation. We explored A. bisporus secretomes and insights obtained can help to better understand biomass valorization.

Neurotransmitter receptors are key molecules in signal transmission, their alterations are associated with brain dysfunction. Relationships between receptors and their corresponding genes are poorly understood, especially in humans. We combined in vitro receptor autoradiography and RNA sequencing to quantify, in the same tissue samples (7 subjects), the densities of 14 receptors and expression levels of their corresponding 43 genes in the Cornu Ammonis (CA) and dentate gyrus (DG) of human hippocampus. Significant differences in receptor densities between both structures were found only for metabotropic receptors, whereas significant differences in RNA expression levels mostly pertained ionotropic receptors. Receptor fingerprints of CA and DG differ in shapes but have similar sizes; the opposite holds true for their “RNA fingerprints”, which represent the expression levels of multiple genes in a single area. In addition, the correlation coefficients between receptor densities and corresponding gene expression levels vary widely and the mean correlation strength was weak-to-moderate. Our results suggest that receptor densities are not only controlled by corresponding RNA expression levels, but also by multiple regionally specific post-translational factors.

Introduction: This mixed-methods study analyzed English-language U.S.-based Twitter posts related to E-cigarette use from February 2021. Methods: Posts were manually identified as health-related or not and, if health-related, whether they were posted by an E-cigarette user. A random selection of 1,000 health-related tweets from 986 unique E-cigarette users were qualitatively content analyzed for theory of planned behavior constructs as well as nature and tone of each tweet message. Using quantitative semantic network analysis, relationships among the identified topics and sentiment-specific conversation patterns were explored. Results: The most salient health-related conversation topics of E-cigarette users, health beliefs corresponding to each theory of planned behavior construct, and major motivational contexts of E-cigarette use were identified. Seven topics emerged in positive tweets: smoking cessation, social impact generation, controls over addiction, therapeutic effects on physical and mental health, social support, device attachment, and peer influence. Nine topics emerged in negative tweets: side effects on physical health, vaping addiction, lack of E-cigarette regulations, peer pressure, increased risk of COVID-19, side effects on mental health, no help in smoking cessation, social conflict, and polysubstance use. Most assertions for E-cigarette benefits were not substantiated. Jokes in tweets appeared to contribute to the view of vaping as an attractive, enjoyable, safe, and fun activity. Discussions about positive aspects of E-cigarette use were concentrated on a few related topics, whereas tweets discouraging E-cigarette use presented a diverse, less related set of topics. Conclusions: The results provide insights into the drivers of E-cigarette use behaviors. E-cigarette user perspectives gathered from social media may inform research to guide future prevention and cessation interventions.

Summary: Arbuscular mycorrhizal fungi (AMF) have evolved associations with roots of 60% plant species, but the net benefit for plants vary broadly from mutualism to parasitism. Yet, we lack a general understanding of the evolutionary and ecological forces driving such variation. To this end, we conducted a comparative phylogenetic experiment with 24 species of Plantago, encompassing worldwide distribution, to address the effect of evolutionary history and environment on plant growth and chemical defenses in response to AMF colonization. We demonstrate that different species within one plant genus vary greatly in their ability to associate with AMF, and that AMF arbuscule colonization intensity decreases monotonically with increasing phylogenetic branch length, but not with concomitant changes in pedological and climatic conditions across species. Moreover, we demonstrate that species with the highest colonization levels are also those that change their defensive chemistry the least. We propose that the costs imposed by high AMF colonization in terms of reduced changes in secondary chemistry might drive the observed macroevolutionary decline in mycorrhization.

We present a 67-year-old male patient who presented with insidious worsening of right hip pain over a 6-month period with clinical and radiographic evidence of severe osteoarthritis. The patient underwent a primary total hip arthroplasty where the femoral head specimen was sent to pathology as a routine specimen. Pathology results demonstrated metastatic adenocarcinoma of prostate origin. The present case emphasizes the importance of routine pathologic examination of femoral head specimens retrieved during total hip arthroplasty, particularly since this was a clinically unsuspected finding. Although cases like these are rare and the process of routine pathologic examination raises a concern for economic implications, a timely diagnosis of adenocarcinoma provides benefits for the patient, for which cost-benefit ratios are difficult to quantify.

Background: Both activated by environmental odorants, there is a clear role for the intranasal trigeminal and olfactory nerves in smell function. Unfortunately, our ability to perceive odorants decreases with age or with injury, and limited interventions are available to treat smell loss. Objective: We investigated whether electrical stimulation of the trigeminal nerve via trigeminal nerve stimulation (TNS) or transcranial direct current stimulation (tDCS) modulates odor sensitivity in healthy individuals. Methods: We recruited 20 healthy adults (12 Female, mean age = 27) to participate in this three-visit, randomized, double-blind, sham-controlled trial. Participants were randomized to receive one of three stimulation modalities (TNS, tDCS, or sham) during each of their visits. Odor detection thresholds were obtained at baseline, immediately post-intervention, and 30-min post-intervention. Furthermore, participants were asked to complete a sustained attention task and mood assessments before odor detection testing. Results: Findings reveal a timeXcondition interaction for guaiacol (GUA) odorant detection thresholds (F (3.188, 60.57) = 3.833, P = 0.0125), but not phenyl ethyl alcohol (PEA) odorant thresholds. At 30-min post-stimulation, both active TNS and active tDCS showed significantly increased sensitivity to GUA compared to sham TNS (Sham TNS = −8.30% vs. Active TNS = 9.11%, mean difference 17.43%, 95% CI 5.674 to 29.18, p = 0.0044; Sham TNS = −8.30% vs. Active tDCS = 13.58%, mean difference 21.89%, 95% CI 10.47 to 33.32, p = 0.0004). Conclusion: TNS is a safe, simple, noninvasive method for boosting olfaction. Future studies should investigate the use of TNS on smell function across different stimulation parameters, odorants, and patient populations.

Background/purpose: A common reason for dental composite restoration failure is recurrent caries at the margins. Our objectives were to: (1) develop a novel low-shrinkage-stress, antibacterial and remineralizing resin composite; (2) evaluate the effects of dimethylaminohexadecyl methacrylate (DMAHDM) on mechanical properties, biofilm inhibition, calcium (Ca) and phosphate (P) ion release, degree of conversion, and shrinkage stress on the new low-shrinkage-stress resin composite for the first time. Material and methods: The resin consisted of urethane dimethacrylate (UDMA) and triethylene glycol divinylbenzyl ether (TEG-DVBE) with high resistance to salivary hydrolytic degradation. Composites were made with 0%–8% of DMAHDM for antibacterial activity, and 20% of nanoparticles of amorphous calcium phosphate (NACP) for remineralization. Mechanical properties and Streptococcus mutans biofilm growth on composites were assessed. Ca and P ion releases, degree of conversion and shrinkage stress were evaluated. Results: Adding 2–5% DMAHDM and 20% NACP into the low-shrinkage-stress composite did not compromise the mechanical properties (p > 0.05). The incorporation of DMAHDM greatly reduced S. mutans biofilm colony-forming units by 2–5 log and lactic acid production by 7 folds, compared to a commercial composite (p

Background: Little is known about the role of disorders of the acromioclavicular joint (ACJ) and how they relate to complications after reverse shoulder arthroplasty (RSA). The purpose of this study is to compare the severity of ACJ osteoarthritis in patients undergoing RSA with and without postoperative acromial and scapular spine fractures. Methods: A retrospective review was performed to identify all patients who underwent primary RSA between 1/1/2007 and 10/31/2019 with a postoperative acromial or scapular spine stress fracture from a single institution. Patients who underwent RSA with a fracture were compared with an age-, sex-, and preoperative diagnosis-matched control group (1:4 controls) with a minimum 2-year follow-up. We compared demographics, medical comorbidities, and ACJ osteoarthritis between the 2 groups. Preoperative radiographs and 3-dimensional computed tomography scans were evaluated for ACJ osteoarthritis in all patients. The Petersson classification, a modified Petersson classification, location of the osteophytes, subchondral cysts, ACJ space, and size of the largest osteophyte were recorded and compared between the 2 groups. Results: The study included 11 patients who underwent primary RSA (8 women and 3 men) with acromial (6) and scapular spine (5) fractures confirmed radiographically and 44 matched controls (average follow-up 3.1 vs. 4.3 years, P = .17). Average age at surgery was similar between study and control groups (69.6 vs. 70.0 years, P = .86). ACJ osteoarthritis with osteophytes larger than 2 mm was common and similar between the 2 groups (91% of patients with acromial fracture and 66% of controls, P = .15). There was no significant difference in the size or location of the ACJ osteophytes. The Petersson classification was similar between groups. However, the percentage of patients with subchondral ACJ cysts was higher in the fracture group (91% vs. 50%, P = .02), and the percentage of patients with large spanning or fused osteophytes was significantly higher in the fracture group (55% vs. 14%, P = .008). Conclusion: Radiographic ACJ osteoarthritis is common in patients undergoing RSA. Severe ACJ osteoarthritis with completely spanning or fused osteophytes may predispose patients to acromial or scapular spine fractures after RSA.

Background: The rate at which patients regain shoulder strength after anatomic and reverse total shoulder arthroplasty (TSA) is unknown. In this study, we aimed to quantify differences in the timeline during which patients gained shoulder strength after primary anatomic and reverse TSA. Methods: We retrospectively reviewed prospectively collected data from 374 shoulders after primary anatomic TSA (aTSA) and 601 shoulders after primary reverse TSA (rTSA). Postoperative improvement in external rotation (ER) strength and forward elevation (FE) strength from baseline was assessed at 3 months, 6 months, 1 year, and 2 years. Percent change in mean shoulder strength between each time point was determined for anatomic and reverse groups separately. A handheld dynamometer was used to assess ER strength with the involved shoulder in 0° ER, 0° abduction, and the elbow in 90° flexion and FE strength with the involved shoulder in the scapular plane at 30° of flexion and 30° of abduction. Results: Both aTSA and rTSA groups ceased to have statistically significant gains in FE strength after 1 year postoperatively. In contrast, patients continued to have statistically significant gains in ER strength between 1 year and 2 years postoperatively after rTSA (P = .001), but not after aTSA (P = .476). Both aTSA and rTSA groups saw improvement in strength in both ER (+32.1% and +51.4%, respectively) and FE (+38.3% and +90.3%, respectively) at 2-year follow-up. The aTSA group’s ER and FE strength increased the most between 3 and 6 months (+16.2% and +35.7%, respectively). In contrast, the rTSA group gained the most ER strength between 6 months and 1 year (+14.8%) and the greatest FE strength between baseline and 3 months (+40.3%). Conclusion: Patients gain ER strength earlier and FE strength later after aTSA compared with rTSA. Most gains in strength occurred in the first year. However, statistically significant gains in shoulder ER strength in the rTSA group continued between 1 year and 2 years postoperatively, suggesting that 2-year follow-up may be inadequate to capture the full benefits of rTSA on shoulder strength. The results of this study provide insight into the timeline of strength recovery after aTSA and rTSA that will help inform patient counseling and future study design.

Background: Reverse total shoulder arthroplasty (RTSA) is a procedure growing in prevalence among younger populations. Consequently, its use in revision arthroplasty is growing in this demographic. However, studies examining the functional outcomes of revision RTSA in younger populations compared with older populations are lacking. The primary purpose of this study is to evaluate the functional outcomes of revision RTSA in patients 65 years old and younger compared with older patients who underwent revision RTSA. We hypothesized that younger patients would have similar outcomes to older patients and both groups would demonstrate improvement in outcomes. Methods: A retrospective review was conducted on a prospectively collected research database at a single tertiary referral center of all patients who underwent RTSA between 2007 and 2018. Patients 65 years old or younger who underwent a revision RTSA and had minimum 2-year follow-up were evaluated. A control group of patients ≥70 years old who underwent revision RTSA were also evaluated. Demographics, surgical factors, active range of motion (ROM), and patient-reported outcomes (PROMs) were compared. The ROM parameters measured were forward elevation, abduction, external rotation, and level of internal rotation. The PROMs collected included American Shoulder and Elbow Surgeons score, Simple Shoulder Test score, University of California–Los Angeles score, Constant score, normalized Constant, and Shoulder Pain and Disability Index 130. The differences in outcomes were compared against the minimal clinically important difference and substantial clinical benefit reported for primary reverse shoulder arthroplasty. Results: A total of 81 patients undergoing revision RTSA were evaluated at a mean follow-up of 4.5 years with 42 patients in the study group and 39 patients in the control group. Both groups demonstrated similar demographics and rates of prior surgeries. Preoperative outcome scores were lower in the study group (≤65 years old) than those in the older control group with American Shoulder and Elbow Surgeons score, Simple Shoulder Test score, and Shoulder Pain and Disability Index 130 remaining worse postoperatively. Both groups experienced statistically significant improvements in ROM from before operation to after operation, with slightly higher improvements in overhead motion in the younger cohort. Both the study group and the control group demonstrated statistically significant improvements in all PROMs with improvement above the substantial clinical benefit for the Constant and Simple Shoulder Test scores. Despite lower functional outcomes reported in the study group postoperatively, the improvement from before operation to after operation in all PROMs was similar between groups. Conclusion: Revision RTSA is a viable option for patients ≤65 years old with a poorly functioning shoulder arthroplasty. ROM and outcome improvements are similar compared with older patients undergoing revision RTSA, but the preoperative and postoperative functional outcomes are worse in the younger patients.

Background: Rotator cuff tears and glenoid loosening remain the two most common causes for revision after anatomic total shoulder arthroplasty. Oversizing of the humeral head leads to increased contact force across the glenohumeral joint and is hypothesized to contribute to clinical and radiographic failure. The purpose of this study is to compare the rate of radiographic overstuffing between standard short humeral heads and newer extra-short heads with decreased lateral offset. Methods: Fifty-five consecutive anatomic total shoulder arthroplasties performed using extra-short humeral heads were retrospectively reviewed and compared with age- and sex-matched controls receiving standard short heads. A total of 110 postoperative radiographs were analyzed using the Iannotti's perfect circle method to compare the prosthesis' center of rotation (COR) with the native humeral head COR. A difference in the COR of >3.0 mm was considered malpositioned. Malpositioning medially was considered overstuffed, and malpositioning laterally was considered understuffed. The direction of displacement of malpositioned prostheses was categorized using a quadrant system. Furthermore, we used a novel method to evaluate medial and superior overstuffing by measuring the displacement between the anatomic and prosthetic head positions along perpendicular axes. Results: Using the Iannotti's perfect circle method, 56% of heads were malpositioned. Overstuffing occurred more frequently with short heads compared with extra-short heads (47% vs. 4%, P

Background: Accurate glenoid component placement in shoulder arthroplasty is often difficult even with the use of preoperative planning. Computer navigation and patient-specific guides increase component placement accuracy, but which patients benefit most is unknown. Our purpose was to assess surgeons' accuracy in placing a glenoid component in vivo using 3-dimensional preoperative planning and standard instruments among various glenoid wear patterns. Methods: We conducted a retrospective review of 170 primary anatomic total shoulder arthroplasty (aTSA) and reverse total shoulder arthroplasty (rTSA) performed at a single institution. Commercially available preoperative planning software was used in all arthroplasties with multiplanar 2-dimensional computed tomography and a 3-dimensional implant overlay. After registration of intraoperative bony landmarks to the navigation system, participating surgeons with knowledge of the preoperative plan were blinded to the computer screen and attempted to implement their preoperative plan by simulating placement of a central-axis glenoid guide pin. Two hundred thirty-three screenshots of surgeon's simulated guide pin placement were included. Glenoid displacement, error in version and inclination, and overall malposition from the preoperatively planned target point were stratified by posterior wear status (with [Walch B2 or B3] or without [A1, A2, or B1]) and Walch classification (A1, A2, B1, B2, or B3). The glenoid component was considered malpositioned when version or inclination errors exceeded 10° or the starting point displacement exceeded 4 mm. Results: For rTSA, errors in version were greater for glenoids with posterior wear compared with those without (8.1° ± 5.6° vs. 4.7° ± 4.0°; P 10° compared with those without (31% vs. 8%; P

Aging is by far the most prominent risk factor for Alzheimer's disease (AD), and both aging and AD are associated with apparent metabolic alterations. As developing effective therapeutic interventions to treat AD is clearly in urgent need, the impact of modulating whole-body and intracellular metabolism in preclinical models and in human patients, on disease pathogenesis, have been explored. There is also an increasing awareness of differential risk and potential targeting strategies related to biological sex, microbiome, and circadian regulation. As a major part of intracellular metabolism, mitochondrial bioenergetics, mitochondrial quality-control mechanisms, and mitochondria-linked inflammatory responses have been considered for AD therapeutic interventions. This review summarizes and highlights these efforts.

Post-acute sequelae of SARS-CoV-2 infection (PASC) affects a wide range of organ systems among a large proportion of patients with SARS-CoV-2 infection. Although studies have identified a broad set of patient-level risk factors for PASC, little is known about the association between “exposome”—the totality of environmental exposures and the risk of PASC. Using electronic health data of patients with COVID-19 from two large clinical research networks in New York City and Florida, we identified environmental risk factors for 23 PASC symptoms and conditions from nearly 200 exposome factors. The three domains of exposome include natural environment, built environment, and social environment. We conducted a two-phase environment-wide association study. In Phase 1, we ran a mixed effects logistic regression with 5-digit ZIP Code tabulation area (ZCTA5) random intercepts for each PASC outcome and each exposome factor, adjusting for a comprehensive set of patient-level confounders. In Phase 2, we ran a mixed effects logistic regression for each PASC outcome including all significant (false positive discovery adjusted p-value < 0.05) exposome characteristics identified from Phase I and adjusting for confounders. We identified air toxicants (e.g., methyl methacrylate), particulate matter (PM2.5) compositions (e.g., ammonium), neighborhood deprivation, and built environment (e.g., food access) that were associated with increased risk of PASC conditions related to nervous, blood, circulatory, endocrine, and other organ systems. Specific environmental risk factors for each PASC condition and symptom were different across the New York City area and Florida. Future research is warranted to extend the analyses to other regions and examine more granular exposome characteristics to inform public health efforts to help patients recover from SARS-CoV-2 infection.

Summary: The extracellular matrix (ECM) plays crucial roles in animal development and diseases. Here, we report that Wnt/β-catenin signaling induces the ECM remodeling during Hydra axis formation. We determined the micro- and nanoscopic arrangement of fibrillar type I collagen along Hydra’s body axis using high-resolution microscopy and X-ray scattering. Elasticity mapping of the ECM ex vivo revealed distinctive elasticity patterns along the body axis. A proteomic analysis of the ECM showed that these elasticity patterns correlate with a gradient-like distribution of metalloproteases along the body axis. Activation of the Wnt/β-catenin pathway in wild-type and transgenic animals alters these patterns toward low ECM elasticity patterns. This suggests a mechanism whereby high protease activity under control of Wnt/β-catenin signaling causes remodeling and softening of the ECM. This Wnt-dependent spatiotemporal coordination of biochemical and biomechanical cues in ECM formation was likely a central evolutionary innovation for animal tissue morphogenesis.

Summary: Protective immune responses against respiratory pathogens, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza virus, are initiated by the mucosal immune system. However, most licensed vaccines are administered parenterally and are largely ineffective at inducing mucosal immunity. The development of safe and effective mucosal vaccines has been hampered by the lack of a suitable mucosal adjuvant. In this study we explore a class of adjuvant that harnesses mucosal-associated invariant T (MAIT) cells. We show evidence that intranasal immunization of MAIT cell agonists co-administered with protein, including the spike receptor binding domain from SARS-CoV-2 virus and hemagglutinin from influenza virus, induce protective humoral immunity and immunoglobulin A production. MAIT cell adjuvant activity is mediated by CD40L-dependent activation of dendritic cells and subsequent priming of T follicular helper cells. In summary, we show that MAIT cells are promising vaccine targets that can be utilized as cellular adjuvants in mucosal vaccines.

Summary: Background: Keratitis ichthyosis deafness (KID) syndrome is a rare disorder caused by hemichannel (HC) activating gain-of-function mutations in the GJB2 gene encoding connexin (Cx) 26, for which there is no cure, or current treatments based upon the mechanism of disease causation. Methods: We applied Adeno Associated Virus (AAV) mediated mAb gene transfer (AAVmAb) to treat the epidermal features of KID syndrome with a well-characterized HC blocking antibody using male mice of a murine model that replicates the skin pathology of the human disease. Findings: We demonstrate that in vivo AAVmAb treatment significantly reduced the size and thickness of KID lesions, in addition to blocking activity of mutant HCs in the epidermis in vivo. We also show that AAVmAb treatment eliminated abnormal keratinocyte proliferation and enlarged cell size, decreased apoptosis, and restored the normal distribution of keratin expression. Interpretation: Our findings reinforce the critical role played by increased HC activity in the skin pathology associated with KID syndrome. They also underscore the clinical potential of anti-HC mAbs coupled with genetic based delivery systems for treating the underlying mechanistic basis of this disorder. Inhibition of HC activity is an ideal therapeutic target in KID syndrome, and the genetic delivery of mAbs targeted against mutant HCs could form the basis of new therapeutic interventions to treat this incurable disease. Funding: Fondazione Telethon grant GGP19148 and University of Padova grant Prot. BIRD187130 to FM; Foundation for Ichthyosis and Related Skin Types (FIRST) and National Institutes of Health grant EY 026911 to TWW.

Summary: Nematocysts are generated by secretion of proteins into a post-Golgi compartment. They consist of a capsule that elongates into a long tube, which is coiled inside the capsule matrix and expelled during its nano-second discharge deployed for prey capture. The driving force for discharge is an extreme osmotic pressure of 150 bar. The complex processes of tube elongation and invagination under these biomechanical constraints have so far been elusive. Here, we show that a non-muscle myosin II homolog (HyNMII) is essential for nematocyst formation in Hydra. In early nematocysts, HyNMII assembles to a collar around the neck of the protruding tube. HyNMII then facilitates tube outgrowth by compressing it along the longitudinal axis as evidenced by inhibitor treatment and genetic knockdown. In addition, live imaging of a NOWA::NOWA-GFP transgenic line, which re-defined NOWA as a tube component facilitating invagination, allowed us to analyze the impact of HyNMII on tube maturation.

Metallic foils are crucial to many applications ranging from printed circuit boards, aviation, aerospace, packaging, photovoltaics to batteries. In these applications they are often part of multifunctional composites, for which mechanical integrity is a prerequisite for the functionality. To ensure precise simulations and lifetime prediction, exact numerical material models are needed. In this work, material parameters of the Lemaitre-Chaboche model were fitted for electrodeposited copper foils. By using nanoindentation and microstructural characterisation, good agreement with literature was achieved. Additionally, it was found that the Tabor rule to estimate yield stress from indentation may not be applicable for metallic foils.

Extensions of the Standard Model have been attempted from the bottom up and from the top down, yet there remains a largely unexplored middle ground. In this paper, using the Mathematica package LieART, we exhaustively enumerate embeddings of the Standard Model within the class of theories with bifundamental fermions in product gauge group SU(a)×SU(b)×SU(c) with no more generators than E6, while achieving SM family unification rather than replication. We incorporate simple phenomenological constraints and find 151 unique models, including 9 that have only vector-like particle content beyond-the-Standard Model (BSM) particles, which we conjecture belong to 5 infinite families of such models. We describe the potentially most viable models: namely, the 9 with strictly vector-like BSM content along with the 29 models we found with no more than 30 additional BSM chiral particles. These include models with fractional electric charge color singlets, and hence magnetic monopoles with multiple Dirac charge. This latter collection of models predicts chiral particles with masses near the electroweak scale accessible to current and future collider experiments.

Summary: Assays detecting blood transcriptome changes are studied for infectious disease diagnosis. Blood-based RNA alternative splicing (AS) events, which have not been well characterized in pathogen infection, have potential normalization and assay platform stability advantages over gene expression for diagnosis. Here, we present a computational framework for developing AS diagnostic biomarkers. Leveraging a large prospective cohort of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and whole-blood RNA sequencing (RNA-seq) data, we identify a major functional AS program switch upon viral infection. Using an independent cohort, we demonstrate the improved accuracy of AS biomarkers for SARS-CoV-2 diagnosis compared with six reported transcriptome signatures. We then optimize a subset of AS-based biomarkers to develop microfluidic PCR diagnostic assays. This assay achieves nearly perfect test accuracy (61/62 = 98.4%) using a naive principal component classifier, significantly more accurate than a gene expression PCR assay in the same cohort. Therefore, our RNA splicing computational framework enables a promising avenue for host-response diagnosis of infection. Motivation: Host-based response assays (HRAs) can often diagnose infectious disease earlier and more precisely than pathogen-based tests. However, the role of RNA alternative splicing (AS) in HRAs remains unexplored, as existing HRAs are restricted to gene expression signatures. We report a computational framework for the identification, optimization, and evaluation of blood AS-based diagnostic assay development for infectious disease. Using SARS-CoV-2 infection as a case study, we demonstrate the improved accuracy of AS biomarkers for COVID-19 diagnosis when compared against six reported transcriptome signatures and when implemented as a microfluidic PCR diagnostic assay.