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Autoantibodies are important for the diagnosis of autoimmune interstitial lung disease (ILD). Standard immunoassays have limitations, including their qualitative nature and/or a narrow dynamic range of detection, hindering the usefulness of autoantibodies as biomarkers of disease activity. Here, the luciferase immunoprecipitation system (LIPS) was evaluated for measuring myositis-specific and other lung-related autoantibodies in 25 subjects with idiopathic inflammatory myopathies (IIM), 26 with Sjögren’s disease (SjD), and 10 healthy volunteers. LIPS detected a broad dynamic range of autoantibodies, to MDA5, Jo-1, PL12, KS, U1-70K, and Ro52, and matched seropositivity status with established immunoassays. Robust anti-MDA5 autoantibodies in four IIM-ILD patients had a median value of 1,134,000 LU (IQR 473,000-2,317,000), which was 500 times higher than in 21 seronegative IIM patients. Markedly elevated anti-Jo-1 autoantibodies in five IIM-ILD patients demonstrated a median value of 1,177,000 LU (IQR: 604,000-2,520,000), which was 1000-fold higher than in seronegative patients. Robust anti-Ro52 and other anti-tRNA-synthetase autoantibodies were detected in a subset of IIM-ILD subjects. In SjD, only anti-U1-70K and KS autoantibodies were identified in ILD patients with a prevalence of 30% and 20%, respectively. In longitudinal samples of five IIM-ILD patients, anti-Jo-1 autoantibody levels paralleled clinical improvement of lung function. LIPS can accurately quantify autoantibody levels as biomarkers for treatment response in patients with autoimmune ILD.

Abstract Advances in gene sequencing technologies have accelerated the identification of genetic variants, but better tools are needed to understand which are causal of disease. This would be particularly useful in fields where gene therapy is a potential therapeutic modality for a disease-causing variant such as inherited retinal disease (IRD). Here, we apply structure-based network analysis (SBNA), which has been successfully utilized to identify variant-constrained amino acid residues in viral proteins, to identify residues that may cause IRD if subject to missense mutation. SBNA is based entirely on structural first principles and is not fit to specific outcome data, which makes it distinct from other contemporary missense prediction tools. In 4 well-studied human disease-associated proteins (BRCA1, HRAS, PTEN, and ERK2) with high-quality structural data, we find that SBNA scores correlate strongly with deep mutagenesis data. When applied to 47 IRD genes with available high-quality crystal structure data, SBNA scores reliably identified disease-causing variants according to phenotype definitions from the ClinVar database. Finally, we applied this approach to 63 patients at Massachusetts Eye and Ear (MEE) with IRD but for whom no genetic cause had been identified. Untrained models built using SBNA scores and BLOSUM62 scores for IRD-associated genes successfully predicted the pathogenicity of novel variants (AUC = 0.851), allowing us to identify likely causative disease variants in 40 IRD patients. Model performance was further augmented by incorporating orthogonal data from EVE scores (AUC = 0.927), which are based on evolutionary multiple sequence alignments. In conclusion, SBNA can used to successfully identify variants as causal of disease in human proteins and may help predict variants causative of IRD in an unbiased fashion.

Abstract Protein-based virus-like particles (P-VLPs) are commonly used to spatially organize antigens and enhance humoral immunity through multivalent antigen display. However, P-VLPs are thymus-dependent antigens that are themselves immunogenic and can induce B cell responses that may neutralize the platform. Here, we investigate thymus-independent DNA origami as an alternative material for multivalent antigen display using the receptor binding domain (RBD) of the SARS-CoV-2 spike protein, the primary target of neutralizing antibody responses. Sequential immunization of mice with DNA-based VLPs (DNA-VLPs) elicits protective neutralizing antibodies to SARS-CoV-2 in a manner that depends on the valency of the antigen displayed and on T cell help. Importantly, the immune sera do not contain boosted, class-switched antibodies against the DNA scaffold, in contrast to P-VLPs that elicit strong B cell memory against both the target antigen and the scaffold. Thus, DNA-VLPs enhance target antigen immunogenicity without generating scaffold-directed immunity and thereby offer an important alternative material for particulate vaccine design.

IntroductionProstate-specific membrane antigen (PSMA) is present in high amounts in salivary glands, but it is unclear whether labeled binders of PSMA are excreted in the saliva.MethodsTen patients with prostate cancer underwent whole-body [18F]DCFPyL PET/CT (NCT03181867), and saliva samples were collected between 0-120 minutes post-injection. [18F]DCFPyL salivary excretion was measured over 120 minutes and expressed as %ID/g. Protein-associated binding was estimated by the percentage of [18F]DCFPyL versus parent radiotracer.ResultsAll PET scans of 10 patients (69 ± 8 years) with histologically confirmed prostate cancer (PSA= 2.4 ± 2.4, and Gleason Grade = 6-9) showed high uptake of [18F]-DCFPyL in salivary glands while 8 patients demonstrated high uptake in the saliva at 45 minutes. The intact [18F]-DCFPyL (98%) was also confirmed in the saliva samples at 120 min with increasing salivary radioactivity between 30-120 min.ConclusionSystemically injected [18F]DCFPyL shows salivary gland uptake, an increasing amount of which is secreted in saliva over time and is not maximized by 120 minutes post-injection. Although probably insignificant for diagnostic studies, patients undergoing PSMA-targeted therapies should be aware of radioactivity in saliva.

Background: There is a gap in the literature regarding musculoskeletal risk factors for concussion within the National Football League (NFL), which is an avenue that must be explored to promote player safety given the high incidence of both injury types. This study aims to observe if ankle injuries are associated with an increased risk of subsequent concussion in NFL players. Methods: The public online database ProFootballReference.com was used to identify ankle injuries and concussions in NFL players from the 2009-2010 to 2019-2020 seasons. Multivariable logistic regression for subsequent concussion and ankle injury was performed, adjusting for body mass index (BMI), age, and player position. For descriptive statistics, unpaired t tests with unequal variance were performed for continuous variables, including BMI and age. χ 2 testing was performed for categorical variables, including player position, and whether the position was offensive, defensive, or on special teams. Results: Of the 5538 NFL players included in the study, 941 had an ankle injury, 633 had a concussion, and 240 had both an ankle injury and a subsequent concussion. The adjusted odds ratio (aOR) for concussion following a single ankle injury was 0.90 (95% CI 0.72-1.14, P = .387); however, the aOR for concussion following multiple ankle injuries was 2.87 (95% CI 1.23-6.75, P = .015). Special teams players had the lowest risk for concussion (aOR 0.17, 95% CI 0.069-0.36, P

Summary: Background: Tracking infectious diseases at the community level is challenging due to asymptomatic infections and the logistical complexities of mass surveillance. Wastewater surveillance has emerged as a valuable tool for monitoring infectious disease agents including SARS-CoV-2 and Mpox virus. However, detecting the Mpox virus in wastewater is particularly challenging due to its relatively low prevalence in the community. In this study, we aim to characterize three molecular assays for detecting and tracking the Mpox virus in wastewater from El Paso, Texas, during February and March 2023. Methods: In this study, a combined approach utilizing three real-time PCR assays targeting the C22L, F3L, and F8L genes and sequencing was employed to detect and track the Mpox virus in wastewater samples. The samples were collected from four sewersheds in the City of El Paso, Texas, during February and March 2023. Wastewater data was compared with reported clinical case data in the city. Findings: Mpox virus DNA was detected in wastewater from all the four sewersheds, whereas only one Mpox case was reported during the sampling period. Positive signals were still observed in multiple sewersheds after the Mpox case was identified. Higher viral concentrations were found in the pellet than in the supernatant of wastewater. Notably, an increasing trend in viral concentration was observed approximately 1–2 weeks before the reporting of the Mpox case. Further sequencing and epidemiological analysis provided supporting evidence for unreported Mpox infections in the city. Interpretation: Our analysis suggests that the Mpox cases in the community is underestimated. The findings emphasize the value of wastewater surveillance as a public health tool for monitoring infectious diseases even in low-prevalence areas, and the need for heightened vigilance to mitigate the spread of Mpox disease for safeguarding global health. Funding: Center of Infectious Diseases at UTHealth, the University of Texas System, and the Texas Epidemic Public Health Institute. The content of this paper is solely the responsibility of the authors and does not necessarily represent the official views of these funding organizations.

ABSTRACT Bacterial infection is the most common complication following staged post-mastectomy breast reconstruction initiated with a tissue expander (TE). To limit bacterial infection, antibiotic irrigation of the surgical site is commonly performed despite little high-quality data to support this practice. We performed a prospective randomized control trial to compare the impact of saline irrigation alone to a triple antibiotic irrigation regimen (1 g cefazolin, 80 mg gentamicin, and 50,000 units of bacitracin in 500 mL of saline) for breast implant surgery. The microbiome in breasts with cancer (n = 16) was compared to those without (n = 16), as all patients (n = 16) had unilateral cancers but bilateral mastectomies (n = 32). Biologic and prosthetic specimens procured both at the time of mastectomy and during TE removal months later were analyzed for longitudinal comparison. Outcomes included clinical infection, bacterial abundance, and relative microbiome composition. No patient in either group suffered a reconstructive failure or developed an infection. Triple antibiotic irrigation administered at the time of immediate TE reconstruction did not reduce bacterial abundance or impact microbial diversity relative to saline irrigation at the time of planned exchange. Implanted prosthetic material adopted the microbial composition of the surrounding host tissue. In cancer-naïve breasts, relative to saline, antibiotic irrigation increased bacterial abundance on periprosthetic capsules (P = 0.03) and acellular dermal matrices (P = 0.04) and altered the microbiota on both. These data show that, relative to saline only, the use of triple antibiotic irrigation in TE breast reconstruction does impact the bacterial abundance and diversity of certain biomaterials from cancer-naïve breasts. IMPORTANCE The lifetime risk of breast cancer is ~13% in women and is treated with a mastectomy in ~50% of cases. The majority are reconstructed, usually starting with a tissue expander to help restore the volume for a subsequent permanent breast implant or the women’s own tissues. The biopsychosocial benefits of breast reconstruction, though, can be tempered by a high complication rate of at least 7% but over 30% in some women. Bacterial infection is the most common complication, and can lead to treatment delays, patient physical and emotional distress and escalating health care cost. To limit this risk, plastic surgeons have tried a variety of strategies to limit bacterial infection including irrigating the pocket created after removing the breast implant with antibiotic solutions, but good-quality data are scarce. Herein, we study the value of antibiotics in pocket irrigation using a robust randomized clinical trial design and molecular microbiology approaches.

Abstract Development of SARS-CoV-2 vaccines that protect vulnerable populations is a public health priority. Here, we took a systematic and iterative approach by testing several adjuvants and SARS-CoV-2 antigens to identify a combination that elicits antibodies and protection in young and aged mice. While demonstrating superior immunogenicity to soluble receptor-binding domain (RBD), RBD displayed as a protein nanoparticle (RBD-NP) generated limited antibody responses. Comparison of multiple adjuvants including AddaVax, AddaS03, and AS01B in young and aged mice demonstrated that an oil-in-water emulsion containing carbohydrate fatty acid monosulphate derivative (CMS:O/W) most effectively enhanced RBD-NP-induced cross-neutralizing antibodies and protection across age groups. CMS:O/W enhanced antigen retention in the draining lymph node, induced injection site, and lymph node cytokines, with CMS inducing MyD88-dependent Th1 cytokine polarization. Furthermore, CMS and O/W synergistically induced chemokine production from human PBMCs. Overall, CMS:O/W adjuvant may enhance immunogenicity and protection of vulnerable populations against SARS-CoV-2 and other infectious pathogens.

Abstract Sjögren’s disease (SjD) is an autoimmune disease that affects exocrine tissues and is characterized by increased apoptosis in salivary and lacrimal glands. Although the pathogenic mechanism triggering SjD is not well understood, overexpression of lysosome-associated membrane protein 3 (LAMP3) is associated with the disease in a subset of SjD patients and the development of SjD-like phenotype in mice. In this study, histological analysis of minor salivary glands of SjD patients suggested that LAMP3-containing material is being ejected from cells. Follow-on in vitro experiments with cells exposed to extracellular particles (EPs) derived from LAMP3-overexpressing cells showed increased apoptosis. Proteomics identified LAMP3 as a major component of EPs derived from LAMP3-overexpressing cells. Live-cell imaging visualized release and uptake of LAMP3-containing EPs from LAMP3-overexpressing cells to naïve cells. Furthermore, experiments with recombinant LAMP3 protein alone or complexed with Xfect protein transfection reagent demonstrated that internalization of LAMP3 was required for apoptosis in a caspase-dependent pathway. Taken together, we identified a new role for extracellular LAMP3 in cell-to-cell communication via EPs, which provides further support for targeting LAMP3 as a therapeutic approach in SjD.

Abstract Sjögren’s disease (SjD) is a chronic autoimmune sialadenitis resulting in salivary gland hypofunction with dry mouth symptom. Previous studies showed that lysosome-associated membrane protein 3 (LAMP3) overexpression is involved in the development of salivary gland hypofunction associated with SjD. However, the molecular mechanisms are still unclear, and no effective treatment exists to reverse gland function in SjD. Analysis on salivary gland samples from SjD patients showed that salivary gland hypofunction was associated with decreased expression of sodium–potassium-chloride cotransporter-1 (NKCC1) and aquaporin 5 (AQP5), which are membrane proteins involved in salivation. Further studies revealed that LAMP3 overexpression decreased their expression levels by promoting endolysosomal degradation. Additionally, we found that LAMP3 overexpression enhanced gene transfer by increasing internalization of adeno-associated virus serotype 2 (AAV2) via the promoted endolysosomal pathway. Retrograde cannulation of AAV2 vectors encoding AQP1 gene (AAV2-AQP1) into salivary glands induced glandular AQP1 expression sufficient to restore salivary flow in LAMP3-overexpressing mice. LAMP3 could play a critical role in the development of salivary gland hypofunction in SjD by promoting endolysosomal degradation of NKCC1 and AQP5. But it also could enhance AAV2-mediated gene transfer to restore fluid movement through induction of AQP1 expression. These findings suggested that AAV2-AQP1 gene therapy is useful in reversing salivary gland function in SjD patients.

Abstract Background Caenorhabditis elegans is an excellent research model whose populations have been used in many studies to address various biological questions. Although worm-to-worm phenotypic variations in isogenic populations have been persistently observed, they are not well understood and are often ignored or averaged out in studies, masking the impacts of such variations on data collection and interpretation. Single-worm RNA sequencing that profiles the transcriptomes of individual animals has the power to examine differences between individuals in a worm population, but this approach has been understudied. The integrity of the starting RNA, the quality of the library and sequence data, as well as the transcriptome-profiling effectiveness of single-worm RNA-seq remain unclear. Therefore, more studies are needed to improve this technique and its application in research. Results In this study, we aimed to develop a single-worm RNA-seq method that includes five steps: worm lysis and RNA extraction, cDNA synthesis, library preparation, sequencing, and sequence data analysis. We found that the mechanical lysis of worms using a Qiagen TissueLyser maintained RNA integrity and determined that the quality of our single-worm libraries was comparable to that of standard RNA-seq libraries based on assessments of a variety of parameters. Furthermore, analysis of pathogen infection-induced gene expression using single-worm RNA-seq identified a core set of genes and biological processes relating to the immune response and metabolism affected by infection. These results demonstrate the effectiveness of our single-worm RNA-seq method in transcriptome profiling and its usefulness in addressing biological questions. Conclusions We have developed a single-worm RNA-seq method to effectively profile gene expression in individual C. elegans and have applied this method to study C. elegans responses to pathogen infection. Key aspects of our single-worm RNA-seq libraries were comparable to those of standard RNA-seq libraries. The single-worm method captured the core set of, but not all, infection-affected genes and biological processes revealed by the standard method, indicating that there was gene regulation that is not shared by all individuals in a population. Our study suggests that combining single-worm and standard RNA-seq approaches will allow for detecting and distinguishing shared and individual-specific gene activities in isogenic populations.

A complete accounting of nearby objects—from the highest-mass white dwarf progenitors down to low-mass brown dwarfs—is now possible, thanks to an almost complete set of trigonometric parallax determinations from Gaia, ground-based surveys, and Spitzer follow-up. We create a census of objects within a Sun-centered sphere of 20 pc radius and check published literature to decompose each binary or higher-order system into its separate components. The result is a volume-limited census of ∼3600 individual star formation products useful in measuring the initial mass function across the stellar (

ABSTRACT Breast implant-associated infections (BIAIs) are the primary complication following placement of breast prostheses in breast cancer reconstruction. Given the prevalence of breast cancer, reconstructive failure due to infection results in significant patient distress and health care expenditures. Thus, effective BIAI prevention strategies are urgently needed. This study tests the efficacy of one infection prevention strategy: the use of a triple antibiotic pocket irrigant (TAPI) against Staphylococcus aureus, the most common cause of BIAIs. TAPI, which consists of 50,000 U bacitracin, 1 g cefazolin, and 80 mg gentamicin diluted in 500 mL of saline, is used to irrigate the breast implant pocket during surgery. We used in vitro and in vivo assays to test the efficacy of each antibiotic in TAPI, as well as TAPI at the concentration used during surgery. We found that planktonically grown S. aureus BIAI isolates displayed susceptibility to gentamicin, cefazolin, and TAPI. However, TAPI treatment enhanced biofilm formation of BIAI strains. Furthermore, we compared TAPI treatment of a S. aureus reference strain (JE2) to a BIAI isolate (117) in a mouse BIAI model. TAPI significantly reduced infection of JE2 at 1 and 7 days postinfection (dpi). In contrast, BIAI strain 117 displayed high bacterial burdens in tissues and implants, which persisted to 14 dpi despite TAPI treatment. Lastly, we demonstrated that TAPI was effective against Pseudomonas aeruginosa reference (PAO1) and BIAI strains in vitro and in vivo. Together, these data suggest that S. aureus BIAI strains employ unique mechanisms to resist antibiotic prophylaxis treatment and promote chronic infection. IMPORTANCE The incidence of breast implant associated infections (BIAIs) following reconstructive surgery postmastectomy remains high, despite the use of prophylactic antibiotic strategies. Thus, surgeons have begun using additional antibiotic-based prevention strategies, including triple antibiotic pocket irrigants (TAPIs). However, these strategies fail to reduce BIAI rates for these patients. To understand why these therapies fail, we assessed the antimicrobial resistance patterns of Staphylococcus aureus strains, the most common cause of BIAI, to the antibiotics in TAPI (bacitracin, cefazolin, and gentamicin). We found that while clinically relevant BIAI isolates were more susceptible to the individual antibiotics compared to a reference strain, TAPI was effective at killing all the strains in vitro. However, in a mouse model, the BIAI isolates displayed recalcitrance to TAPI, which contrasted with the reference strain, which was susceptible. These data suggest that strains causing BIAI may encode specific recalcitrance mechanisms not present within reference strains.

Interleukin (IL)-7 is broadly active on T-cell populations, and modified versions have been clinically evaluated for a variety of therapeutic applications, including cancer, lymphopenia, and infectious diseases; and found to be relatively well-tolerated and biologically active. Here we describe novel IL-7R agonists that are unrelated in structure to IL-7, bind to the receptor subunits differently from IL-7, but closely emulate IL-7 biology. The small size, low structural complexity, and the natural amino acid composition of the pharmacologically active peptide MDK1472 allows facile incorporation into protein structures, such as the IgG2-Fc fusion MDK-703. This molecule possesses properties potentially better suited to therapeutic applications than native IL-7 or its derivatives. We compared these compounds with IL-7 for immune cell selectivity, induction of IL-7R signaling, receptor-mediated internalization, proliferation, and generation of immune cell phenotypes in human and non-human primate (NHP) peripheral blood cells in vitro; and found them to be similar in biological activity to IL-7. In cynomolgus macaques, MDK-703 exhibits a circulating half-life of 46 hr and produces sustained T-cell expansion characteristic of IL-7 treatment. In the huCD34+-engrafted NSG mouse model of the human immune system, MDK-703 induces an immune cell profile very similar to that generated by IL-7-derived compounds; including the pronounced expansion of memory T-cells, particularly the population of stem-like memory T-cells (Tscm) which may be important for anti-tumor activities reported with IL-7 treatment. Clinical administration of IL-7 and modified variants has been reported to induce anti-drug antibodies (ADAs), including IL-7 neutralizing antibodies. The novel peptide agonist reported here scores very low in predicted immunogenicity, and because the peptide lacks sequence similarity with IL-7, the problematic immunogenic neutralization of endogenous cytokine should not occur. The properties we report here implicate MDK-703 as a candidate for clinical evaluation in oncology, anti-viral and other infectious disease, vaccine enhancement, and treatment of lymphopenia.

The recent development of a system for long-term in vitro culture of the syphilis spirochete, Treponema pallidum subsp. pallidum, has introduced the possibility of detailed genetic analysis of this bacterium. In this study, the in vitro culture system was used to isolate and characterize clonal populations of T. pallidum subsp. pallidum Nichols, the most widely studied strain. In limiting dilutions experiments, it was possible to establish cultures with inocula as low as 0.5 T. pallidum per well despite the long generation time (~35 to 40 hours) of this organism. Six Nichols strain clones isolated by limiting dilution were characterized in detail. All clones exhibited indistinguishable morphology and motility, highly similar in vitro multiplication rates, and comparable infectivity in the rabbit model (ID50 ≤ 100 bacteria). Genomic sequencing revealed sequence heterogeneity in the form of insertions or deletions at 5 sites, single nucleotide variations at 20 sites, and polynucleotide (polyG/C) tract length differences at 22 locations. Genomic sequences of the uncloned Nichols strain preparations propagated in rabbits or in vitro cultures exhibited substantial heterogeneity at these locations, indicating coexistence of many varied 'clonotypes' within these populations. Nearly all genetic variations were specific for the Nichols strain and were not detected in the >280 T. pallidum genomic sequences that are currently available. We hypothesize that these Nichols strain-specific sequence variations arose independently either during human infection or within the 110 years since the strain's initial isolation, and thus represent examples of microevolution and divergence.

Hydroxychloroquine (HCQ) is a lysosomotropic agent that is commonly used for treating Sjögren's disease (SjD). However, its efficacy is controversial because of the divergent response to the drug among patients. In a subgroup of SjD patients, lysosome-associated membrane protein 3 (LAMP3) is elevated in expression in the salivary glands and promotes lysosomal dysregulation and lysosome-dependent apoptotic cell death. In this study, chloroquine (CQ) and its derivative HCQ were tested for their ability to prevent LAMP3-induced apoptosis, in vitro and on a mouse model of SjD. In addition, efficacy of HCQ treatment was retrospectively compared between high LAMP3 mRNA expression in minor salivary glands and those with LAMP3 mRNA levels comparable with healthy controls. Study results show that CQ treatment stabilized the lysosomal membrane in LAMP3-overexpressing cells via deactivation of cathepsin B, resulting in decreased apoptotic cell death. In mice with established SjD-like phenotype, HCQ treatment also significantly decreased apoptotic cell death and ameliorated salivary gland hypofunction. Retrospective analysis of SjD patients found that HCQ tended to be more effective in improving disease activity index, symptom severity and hypergammaglobulinemia in patients with high LAMP3 expression compared those with normal LAMP3 expression. Taken together, these findings suggested that by determining salivary gland LAMP3 mRNA level, a patient's response to HCQ treatment could be predicted. This finding may provide a novel strategy for guiding the development of more personalized medicine for SjD.

Domestic wastewater, when collected and evaluated appropriately, can provide valuable health-related information for a community. As a relatively unbiased and non-invasive approach, wastewater surveillance may complement current practices towards mitigating risks and protecting population health. Spurred by the COVID-19 pandemic, wastewater programs are now widely implemented to monitor viral infection trends in sewersheds and inform public health decision-making. This review summarizes recent developments in wastewater-based epidemiology for detecting and monitoring communicable infectious diseases, dissemination of antimicrobial resistance, and illicit drug consumption. Wastewater surveillance, a quickly advancing Frontier in environmental science, is becoming a new tool to enhance public health, improve disease prevention, and respond to future epidemics and pandemics.

Benzodiazepines (BZDs) are among one of the most widely prescribed drug classes in the United States. BZDs are a class of psychoactive drugs known for their depressant effect on the central nervous system (CNS). They quickly diffuse through the blood–brain barrier to affect the inhibitory neurotransmitter GABA and exert sedative effects. Related to their rapid onset and immediate symptom relief, BZDs are used for those struggling with sleep, anxiety, spasticity due to CNS pathology, muscle relaxation, and epilepsy. One of the debilitating side effects of BZDs is their addictive potential. The dependence on BZDs generally leads to withdrawal symptoms, requiring careful tapering of the medication when prescribed. Regular use of BZDs has been shown to cause severe, harmful psychological and physical dependence, leading to withdrawal symptoms similar to that of alcohol withdrawal. Some of these withdrawal symptoms can be life threatening. The current treatment for withdrawal is through tapering with clonazepam. Many drugs have been tested as a treatment for withdrawal, with few proving efficacious in randomized control trials. Future research is warranted for further exploration into alternative methods of treating BZD withdrawal. This call to action proves especially relevant, as those seeking treatment for BZD dependence and withdrawal are on the rise in the United States.

Abstract Machine learning algorithms are being increasingly used to process large volumes of wildlife imagery data from unmanned aerial vehicles (UAVs); however, suitable algorithms to monitor multiple species are required to enhance efficiency. Here, we developed a machine learning algorithm using a low‐cost computer. We trained a convolutional neural network and tested its performance in: (1) distinguishing focal organisms of three marine taxa (Australian fur seals, loggerhead sea turtles and Australasian gannets; body size ranges: 0.8–2.5 m, 0.6–1.0 m, and 0.8–0.9 m, respectively); and (2) simultaneously delineating the fine‐scale movement trajectories of multiple sea turtles at a fish cleaning station. For all species, the algorithm performed best at detecting individuals of similar body length, displaying consistent behaviour or occupying uniform habitat (proportion of individuals detected, or recall of 0.94, 0.79 and 0.75 for gannets, seals and turtles, respectively). For gannets, performance was impacted by spacing (huddling pairs with offspring) and behaviour (resting vs. flying shapes, overall precision: 0.74). For seals, accuracy was impacted by morphology (sexual dimorphism and pups), spacing (huddling and creches) and habitat complexity (seal sized boulders) (overall precision: 0.27). For sea turtles, performance was impacted by habitat complexity, position in water column, spacing, behaviour (interacting individuals) and turbidity (overall precision: 0.24); body size variation had no impact. For sea turtle trajectories, locations were estimated with a relative positioning error of

The purpose of this study was to use the Altmetric Attention Score to determine the 50 most impactful medial ulnar collateral ligament articles in online media and compare their characteristics to the most-cited medial ulnar collateral ligament articles in the scientific literature. The Altmetric database was queried to identify all published articles about the medial ulnar collateral ligament, and this list was stratified by the Altmetric Attention Score to identify the 50 highest scoring articles. Several data elements were extracted, including article topic, article type, journal name, and the number of online mentions on Facebook, Twitter, news, and other platforms. Each article’s geographic origin was determined based on the institutional affiliation of the first author. Our index search yielded 1283 articles published between 1987 and 2020, from which the 50 articles with the highest Altmetric Attention Scores were included for analysis. Altmetric Attention Scores of the top 50 medial ulnar collateral ligament articles ranged from 20 to 482 (median: 32, interquartile range: 20–62). The most common article type was original research (72%), and the most common topic was epidemiology/risk factors (26%). A majority of studies were Level 3 (36%) or Level 4 evidence (36%). Of the top 50 medial ulnar collateral ligament articles, 94% originated from the United States. A few articles had a high Altmetric Attention Score, suggesting that medial ulnar collateral ligament research does not generate consistently high online attention. The lack of Level 1 studies suggests the need for high-level studies on the medial ulnar collateral ligament. Most studies originated in the United States and were published in the American Orthopaedic Society for Sports Medicine–affiliated journals. The medial ulnar collateral ligament articles included in this study differed substantially from a previous report of the most-cited medial ulnar collateral ligament articles in the literature, suggesting that alternative metrics add a unique dimension to understanding the overall impact of published research on the medial ulnar collateral ligament.

Remotely piloted aircraft systems (RPAS, or ‘drones’ hereafter) have potential for surveying waterbird species and habitats, but there is a risk that the disturbance from drones could compromise count accuracy and bird welfare. We examined the response of 16 waterbird species to repeated up-and-back overhead drone flights (n = 50 flights) at multiple flight heights (80, 60, 40 and 20 m) using three common drone platforms (DJI Matrice 300, DJI Mavic 2 Enterprise Advanced and DJI Phantom 4). A ground observer scored the species’ responses to overhead drone flights, which ranged from no response (no change to initial behavior), vigilance (head turning and tracking), movement within the site (swimming, diving, flight into or on the water) and substantial flight resulting in departure from the pond (fleeing). A total of 280 waterbird encounters with overhead drones were observed. The most common response across all flights was no response (70.7%), followed by vigilance (27.5%), whereas more intense responses were comparatively rare (1.8%). The responses were of higher intensity during earlier overhead drone flights, before moderating substantially during later flights. Thus, our case study provides the first unambiguous evidence of the attenuation of responses of bird species to drones.

Abstract Background PSMA-targeted radionuclide therapy (TRT) is a promising treatment for prostate cancer (PCa), but dose-limiting xerostomia can severely limit its clinical adaptation, especially when using alpha-emitting radionuclides. With [18F]DCFPyL as a surrogate for PSMA-TRT, we report a novel method to selectively reduce salivary gland (SG) uptake of systemically administered [18F]DCFPyL by immediate prior infusion of non-radioactive standard of [18F]DCFPyL (DCFPyL) directly into the SG via retrograde cannulation. Methods A dose-finding cohort using athymic nude mice demonstrated proof of principle that SG uptake can be selectively blocked by DCFPyL administered either locally via cannulation (CAN group) or systemically (SYS group). The experiments were repeated in a validation cohort of 22RV1 tumor-bearing mice. Submandibular glands (SMG) of CAN mice were locally blocked with either saline or DCFPyL (dose range: 0.01× to 1000× molar equivalent of the radioactive [18F]DCFPyL dose). The radioactive dose of [18F]DCFPyL was administered systemically 10 min later and the mice euthanized after 1 h for biodistribution studies. Toxicity studies were done at up to 1000× dose. Results In the dose-finding cohort, the SYS group showed a dose-dependent 12–40% decrease in both the SMG T/B and the kidney (tumor surrogate). Mild blocking was observed at 0.01× , with maximal blocking reached at 1× with no additional blocking up to 1000× . In the CAN group, blocking at the 0.1× and 1× dose levels resulted in a similar 42–53% decrease, but without the corresponding decrease in kidney uptake as seen in the SYS group. Some evidence of “leakage” of DCFPyL from the salivary gland into the systemic circulation was observed. However, experiments in 22RV1 tumor-bearing mice at the 0.1× and 1× dose levels confirm that, at the appropriate blocking dose, SG uptake of [18F]DCFPyL can be selectively reduced without affecting tumor uptake and with no toxicity. Conclusion Our results suggest that direct retrograde instillation of DCFPyL into the SG could predictably and selectively decrease salivary uptake of systemically administered [18F]DCFPyL without altering tumor uptake, if given at the appropriate dose. This novel approach is easily translatable to clinical practice and has the potential to mitigate xerostomia, without compromising the therapeutic efficacy of the PSMA-TRT.

Since the emergence of SARS-CoV-2 (SARS-2), multiple vaccine candidates were developed and studied both preclinically and clinically. Nearly all are based on the SARS-2 spike glycoprotein or its receptor binding domain (RBD). Studies of these vaccine candidates have largely been in a SARS-2 naïve context. However, pre-existing immunity to SARS-2 acquired through infection or vaccination continues to increase. Evaluating future vaccine candidates in context of this pre-existing immunity is necessary to understand how immune responses are subsequently influenced. Here, we evaluated the serum and IgG+ B cell responses to the SARS-2 RBD in context of pre-existing immunity elicited by the full SARS-2 spike, and we compared this to boosting with the full SARS-2 spike. Boosting with the SARS-2 RBD resulted in increased reactivity to RBD epitopes, but both immunization regimens resulted in similarly broad neutralization across diverse sarbecoviruses. These findings may inform comparison among SARS-2 RBD-based vaccine candidates to currently approved spike-based candidates.

Current standards for safe delivery of electrical stimulation to the central nervous system are based on foundational studies which examined post-mortem tissue for histological signs of damage. This set of observations and the subsequently proposed limits to safe stimulation, termed the “Shannon limits,” allow for a simple calculation (using charge per phase and charge density) to determine the intensity of electrical stimulation that can be delivered safely to brain tissue. In the three decades since the Shannon limits were reported, advances in molecular biology have allowed for more nuanced and detailed approaches to be used to expand current understanding of the physiological effects of stimulation. Here, we demonstrate the use of spatial transcriptomics (ST) in an exploratory investigation to assess the biological response to electrical stimulation in the brain. Electrical stimulation was delivered to the rat visual cortex with either acute or chronic electrode implantation procedures. To explore the influence of device type and stimulation parameters, we used carbon fiber ultramicroelectrode arrays (7 μm diameter) and microwire electrode arrays (50 μm diameter) delivering charge and charge density levels selected above and below reported tissue damage thresholds (range: 2–20 nC, 0.1–1 mC/cm2). Spatial transcriptomics was performed using Visium Spatial Gene Expression Slides (10x Genomics, Pleasanton, CA, United States), which enabled simultaneous immunohistochemistry and ST to directly compare traditional histological metrics to transcriptional profiles within each tissue sample. Our data give a first look at unique spatial patterns of gene expression that are related to cellular processes including inflammation, cell cycle progression, and neuronal plasticity. At the acute timepoint, an increase in inflammatory and plasticity related genes was observed surrounding a stimulating electrode compared to a craniotomy control. At the chronic timepoint, an increase in inflammatory and cell cycle progression related genes was observed both in the stimulating vs. non-stimulating microwire electrode comparison and in the stimulating microwire vs. carbon fiber comparison. Using the spatial aspect of this method as well as the within-sample link to traditional metrics of tissue damage, we demonstrate how these data may be analyzed and used to generate new hypotheses and inform safety standards for stimulation in cortex.

Objectives: To identify the top 100 most impactful anterior cruciate ligament articles in online media as measured by the Altmetric Attention Score and compare their characteristics to the most-cited anterior cruciate ligament articles in the scientific literature. Methods: The Altmetric database was queried to identify all published articles pertaining to the anterior cruciate ligament. The search yielded 9445 articles, which were stratified by highest to lowest Altmetric Attention Score. The top 100 articles were included. Collected data included article type, article topic, journal name, and online mentions in news, blogs, Twitter, Facebook, Wikipedia, and other sources. The geographic origin of each article was also determined based on the institutional affiliation of the first author. Results: Altmetric Attention Score of the top 100 anterior cruciate ligament articles ranged from 109 to 2193 (median 172.0, interquartile range 137.5–271.5). Of the 100 articles, 65 were published in three journals: American Journal of Sports Medicine, British Journal of Sports Medicine, and Journal of Orthopaedic & Sports Physical Therapy. The most prevalent article type was original research (60%), followed by systematic review/meta-analysis (18%). The most prevalent article topic was rehabilitation and return to play after anterior cruciate ligament reconstruction (22%), followed by epidemiology/risk factors (16%), injury prevention (14%), and biomechanics of anterior cruciate ligament injuries (14%). Of the top 100 articles, 54% were American, 31% were European, and 15% were published in other countries outside of the United States and Europe. Conclusion: This study used Altmetric Attention Score to identify the 100 most engaged anterior cruciate ligament articles in online media. The characteristics of these articles differed substantially from the most-cited anterior cruciate ligament articles in the literature with regard to article type, article topic, geographic origin, and publication journal. These findings suggest that alternative metrics measure distinct components of anterior cruciate ligament article engagement and add an important dimension to understanding the overall impact of published research on the anterior cruciate ligament.

Abstract Several early childhood obesity prediction models have been developed, but none for New Zealand's diverse population. We aimed to develop and validate a model for predicting obesity in 4–5-year-old New Zealand children, using parental and infant data from the Growing Up in New Zealand (GUiNZ) cohort. Obesity was defined as body mass index (BMI) for age and sex ≥ 95th percentile. Data on GUiNZ children were used for derivation (n = 1731) and internal validation (n = 713). External validation was performed using data from the Prevention of Overweight in Infancy Study (POI, n = 383) and Pacific Islands Families Study (PIF, n = 135) cohorts. The final model included: birth weight, maternal smoking during pregnancy, maternal pre-pregnancy BMI, paternal BMI, and infant weight gain. Discrimination accuracy was adequate [AUROC = 0.74 (0.71–0.77)], remained so when validated internally [AUROC = 0.73 (0.68–0.78)] and externally on PIF [AUROC = 0.74 [0.66–0.82)] and POI [AUROC = 0.80 (0.71–0.90)]. Positive predictive values were variable but low across the risk threshold range (GUiNZ derivation 19–54%; GUiNZ validation 19–48%; and POI 8–24%), although more consistent in the PIF cohort (52–61%), all indicating high rates of false positives. Although this early childhood obesity prediction model could inform early obesity prevention, high rates of false positives might create unwarranted anxiety for families.

Background: Lateral patellar instability is common in young, active patients. Patients who fail conservative treatment may benefit from medial patellofemoral ligament (MPFL) reconstruction. Indications: Recurrent lateral patellar instability. Technique Description: Examination assesses patellar translation, eversion and range of instability. Diagnostic arthroscopy is performed for loose bodies, cartilage damage, trochlear dysplasia, and tracking. A longitudinal incision is made from superomedial to mid-body of the patella. The plane between the capsule and retinaculum is developed for palpation of the medial epicondyle and adductor tubercle. Electrocautery and rongeur are used to create a trough on the patella from centromedially to superomedially. Two suture anchors are placed at the proximal and distal trough. Fluoroscopy is utilized to identify Schöttle point with a perfect lateral radiograph. A 3-centimeter incision is made, and blunt dissection connects the 2 incisions in the developed plane. A guidepin is advanced at Schöttle point, and suture anchor sutures are shuttled through the plane, posterior to the guidepin. There should be loosening of tension with knee flexion. A semitendinosus allograft is whipstitched with terminal tapering. The whipstitched end is tunneled around the guidepin and brought back to the patella, so that both ends have an excess of 20 to 25 millimeters, and excess graft is trimmed from the free end prior to whipstitching. The doubled graft is sized. The midportion of the tendon is tagged and passed through both anchors. The graft is tensioned to the patellar trough and the graft ends are advanced through the developed plane. The femoral tunnel is reamed, and the graft is tensioned into the tunnel after nitinol wire placement with the knee in full extension. Isometry and lateral patellar translation are assessed, aiming for 1 quadrant with firm endpoint. The patella is proximalized and the tensioned graft is secured with an interference screw. Results: MPFL reconstruction is successful for the majority of patients, with 1.2% reporting instability, 3.6% apprehension, and 3.1% reoperation. Possible complications include patellar fracture, patellofemoral pain, and knee stiffness (loss of range of motion). Discussion/Conclusion: Lateral patellar instability is common, and MPFL reconstruction is typically successful for the majority of patients.

ABSTRACT Carbapenem-resistant Klebsiella pneumoniae (CRKp) is an urgent public health threat. Worldwide dissemination of CRKp has been largely attributed to clonal group (CG) 258. However, recent evidence indicates the global emergence of a CRKp CG307 lineage. Houston, TX, is the first large city in the United States with detected cocirculation of both CRKp CG307 and CG258. We sought to characterize the genomic and clinical factors contributing to the parallel endemic spread of CG258 and CG307. CRKp isolates were collected as part of the prospective, Consortium on Resistance against Carbapenems in Klebsiella and other Enterobacterales 2 (CRACKLE-2) study. Hybrid short-read and long-read genome assemblies were generated from 119 CRKp isolates (95 originated from Houston hospitals). A comprehensive characterization of phylogenies, gene transfer, and plasmid content with pan-genome analysis was performed on all CRKp isolates. Plasmid mating experiments were performed with CG307 and CG258 isolates of interest. Dissection of the accessory genomes suggested independent evolution and limited horizontal gene transfer between CG307 and CG258 lineages. CG307 contained a diverse repertoire of mobile genetic elements, which were shared with other non-CG258 K. pneumoniae isolates. Three unique clades of Houston CG307 isolates clustered distinctly from other global CG307 isolates, indicating potential selective adaptation of particular CG307 lineages to their respective geographical niches. CG307 strains were often isolated from the urine of hospitalized patients, likely serving as important reservoirs for genes encoding carbapenemases and extended-spectrum β-lactamases. Our findings suggest parallel cocirculation of high-risk lineages with potentially divergent evolution. IMPORTANCE The prevalence of carbapenem-resistant Klebsiella pneumoniae (CRKp) infections in nosocomial settings remains a public health challenge. High-risk clones such as clonal group 258 (CG258) are particularly concerning due to their association with blaKPC carriage, which can severely complicate antimicrobial treatments. There is a recent emergence of clonal group 307 (CG307) worldwide with little understanding of how this successful clone has been able to adapt while cocirculating with CG258. We provide the first evidence of potentially divergent evolution between CG258 and CG307 with limited sharing of adaptive genes. Houston, TX, is home to the largest medical center in the world, with a large influx of domestic and international patients. Thus, our unique geographical setting, where two pandemic strains of CRKp are circulating, provides an indication of how differential accessory genome content can drive stable, endemic populations of CRKp. Pan-genomic analyses such as these can reveal unique signatures of successful CRKp dissemination, such as the CG307-associated plasmid (pCG307_HTX), and provide invaluable insights into the surveillance of local carbapenem-resistant Enterobacterales (CRE) epidemiology.

BMP6 is a central cytokine in the induction of Sjögren’s syndrome–associated (SS-associated) secretory hypofunction. However, the upstream initiation leading to the production of this cytokine in SS is unknown. In this study, RNA ISH on salivary gland sections taken from patients with SS indicated monocytic lineage cells as a cellular source of BMP6. RNA-Seq data on human salivary glands suggested that TLR4 signaling was an upstream regulator of BMP6, which was confirmed by in vitro cell assays and single-cell transcriptomics of human PBMCs. Further investigation showed that HSP70 was an endogenous natural TLR4 ligand that stimulated BMP6 expression in SS. Release of HSP70 from epithelial cells could be triggered by overexpression of lysosome-associated membrane protein 3 (LAMP3), a protein also associated with SS in several transcriptome studies. In vitro studies supported the idea that HSP70 was released as a result of lysosomal exocytosis initiated by LAMP3 expression, and reverse transcription PCR on RNA from minor salivary glands of patients with SS confirmed a positive correlation between BMP6 and LAMP3 expression. BMP6 expression could be experimentally induced in mice by overexpression of LAMP3, which developed an SS-like phenotype. The newly identified LAMP3/HSP70/BMP6 axis provided an etiological model for SS gland dysfunction and autoimmunity.

The antibody profile against autoantigens previously associated with autoimmune diseases and other human proteins in patients with COVID-19 or multisystem inflammatory syndrome in children (MIS-C) remains poorly defined. Here we show that 30% of adults with COVID-19 had autoantibodies against the lung antigen KCNRG, and 34% had antibodies to the SLE-associated Smith-D3 protein. Children with COVID-19 rarely had autoantibodies; one of 59 children had GAD65 autoantibodies associated with acute onset of insulin-dependent diabetes. While autoantibodies associated with SLE/Sjögren’s syndrome (Ro52, Ro60, and La) and/or autoimmune gastritis (gastric ATPase) were detected in 74% (40/54) of MIS-C patients, further analysis of these patients and of children with Kawasaki disease (KD), showed that the administration of intravenous immunoglobulin (IVIG) was largely responsible for detection of these autoantibodies in both groups of patients. Monitoring in vivo decay of the autoantibodies in MIS-C children showed that the IVIG-derived Ro52, Ro60, and La autoantibodies declined to undetectable levels by 45-60 days, but gastric ATPase autoantibodies declined more slowly requiring >100 days until undetectable. Further testing of IgG and/or IgA antibodies against a subset of potential targets identified by published autoantigen array studies of MIS-C failed to detect autoantibodies against most (16/18) of these proteins in patients with MIS-C who had not received IVIG. However, Troponin C2 and KLHL12 autoantibodies were detected in 2 of 20 and 1 of 20 patients with MIS-C, respectively. Overall, these results suggest that IVIG therapy may be a confounding factor in autoantibody measurements in MIS-C and that antibodies against antigens associated with autoimmune diseases or other human proteins are uncommon in MIS-C.

Summary: Eliciting antibodies to surface-exposed viral glycoproteins can generate protective responses that control and prevent future infections. Targeting conserved sites may reduce the likelihood of viral escape and limit the spread of related viruses with pandemic potential. Here we leverage rational immunogen design to focus humoral responses on conserved epitopes. Using glycan engineering and epitope scaffolding in boosting immunogens, we focus murine serum antibody responses to conserved receptor binding motif (RBM) and receptor binding domain (RBD) epitopes following severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike imprinting. Although all engineered immunogens elicit a robust SARS-CoV-2-neutralizing serum response, RBM-focusing immunogens exhibit increased potency against related sarbecoviruses, SARS-CoV, WIV1-CoV, RaTG13-CoV, and SHC014-CoV; structural characterization of representative antibodies defines a conserved epitope. RBM-focused sera confer protection against SARS-CoV-2 challenge. Thus, RBM focusing is a promising strategy to elicit breadth across emerging sarbecoviruses without compromising SARS-CoV-2 protection. These engineering strategies are adaptable to other viral glycoproteins for targeting conserved epitopes.

Background: As the popularity of Osteochondral allograft (OCA) transplantation increases, studies have commonly investigated potential risk factors that may influence OCA transplantation survivorship. Objectives: To investigate which risk factors of OCA transplantation failure have been evaluated, which have been found to be significant, and the frequency of inclusion of identified risk factors. Data sources: EMBASE, OVID, and Medline Study eligibility criteria: Studies investigating failure after OCA transplantation comparing or investigating one or more risk factors for failure through any statistical approach. Study appraisal and synthesis: All investigated failure risk factors and associated statistical significance were extracted and tabulated by study. The frequency of significance of each factor and inclusiveness of risk factor analysis by each study were assessed. Results: A total of 15 studies were included for qualitative analysis. Age, sex, body mass index (BMI), previous surgeries, and lesion location were among the most frequently investigated variables. Only BMI and previous surgeries were significant in more than one study. Limitations: There was heterogeneity in surgical techniques and varying definitions of failure utilized by each study, making a direct comparison difficult. Conclusions and implications of key findings: OCA failure risk factor analyses are limited primarily to basic demographic factors (eg, age, sex). Furthermore, only 2 identified risk factors were significant in more than one study, limiting the ability to make evidence-based decisions with the current data available. This information could impact operative planning, the decision to perform an OCA over other cartilage restoration procedures, and patient expectations. Systematic review registration number: CRD42021282852

Baseball is unique in its multiple facets: pitching, hitting, base rounding, and fielding are distinct activities that require different athletic skills to perform at a high level. Likewise, these different aspects of the game can contribute to a multitude of varying injuries. While high-velocity overhead throwing, along with batting, can produce a plethora of upper extremity injuries that often garner attention, injuries to the lower extremity can severely impact a player’s performance and ability to compete. The rigors of the short, explosive sprinting required for base running, as well as the dynamic movement required for fielding, create ample opportunity for lower limb injury, and even subtle pathology can affect a pitcher’s ability to perform or increase their long-term risk of injury. Chronic injury from conditions such as femoroacetabular impingement and hip labral tears can also occur. The purpose of the present review is to summarize the relevant epidemiology, pathophysiology, and treatment of lower extremity injuries in baseball athletes, with reference to current research into the prevention and management of such injuries.

Abstract Purpose The purpose of our study was to use the Altmetric Attention Score (AAS) to evaluate the 100 most impactful articles in online media pertaining to the rotator cuff and compare their characteristics to the most‐cited rotator cuff articles in the scientific literature. Methods We performed an article extraction using Altmetric Explorer to identify all published articles pertaining to the rotator cuff. The top 100 articles with the highest AAS were included for analysis. Several data elements were extracted for each included article: title, article type, article topic, year of publication, journal name, authors, institutional affiliations, and online mentions (i.e. the number of times the article was mentioned in news, blog, Twitter, Facebook, and Wikipedia sources). The geographic origin of each article was also determined by the institutional affiliation of the first author, which was categorized as American (originating in the United States), European (originating in Europe), or other. Results The 100 articles with the highest AAS were published between 2009 and 2020, with AAS ranging from 47 to 676 (median: 74.5, 25th percentile: 59.5, 75th percentile: 114.5). Of all online media sources, Twitter correlated most strongly with AAS (r = 0.9007, r2 = 0.8112). The selected articles were most frequently published in the American Journal of Sports Medicine (13), the Journal of Shoulder and Elbow Surgery (11), and the Journal of Bone and Joint Surgery (7). The most common article type was Systematic Review/Meta‐Analysis (29%), followed by Randomized Controlled Trial (15%). The top 3 AAS articles were all published by authors based in Europe. Conclusion The most impactful rotator cuff articles in online media generated substantial online attention. These studies were often performed in Europe and tended to be high level of evidence, focusing on treatment of rotator cuff pathology. The rotator cuff articles that produced the most online attention differed from a previous report of the most‐cited rotator cuff articles, suggesting that alternative metrics may be used in concert with conventional bibliometrics to obtain a more complete representation of scientific impact.

Category: Ankle; Hindfoot; Sports Introduction/Purpose: Achilles tendon injuries affect a wide range of individuals, including both recreational and competitive athletes. In the elite athlete population, Achilles tendon ruptures are typically season-ending injuries and can be career-threatening. Due to these impacts on individual players' careers as well as team success, Achilles tendon injuries often generate substantial discussion in the mainstream media and among the general public. However, traditional citation-based metrics fail to capture the dissemination of Achilles research that occurs outside the realm of scientific journal publications. Thus, the purpose of this study was to use the Altmetric Attention Score (AAS) to identify the 100 most impactful articles in online media pertaining to the Achilles tendon and assess their characteristics. Methods: The Altmetric database (Digital Science, Holtzbrinck Publishing) was queried to identify all published articles pertaining to the Achilles Tendon. The articles were stratified by highest to lowest AAS and the top 100 articles with the highest scores were included for analysis. Several data elements were extracted for each article: title, article type, article topic, year of publication, journal name, authors, institutional affiliations, and online mentions (i.e., the number of times the article was mentioned in news, blog, Twitter, Facebook and Wikipedia sources). The geographic origin of each article was also determined by the institutional affiliation of the first author, which was categorized as American (originating in the United States), European (originating in Europe), or Other. Linear regression was used to determine the relationship between online mentions and AAS. Results: The initial search yielded 3,810 articles published between 1957 and 2021. AAS of the top 100 articles ranged from 37 to 476 with a median of 65 (interquartile range: 42-110). The selected articles were published in 39 journals. The most prevalent article type was randomized controlled trial (21.8%), followed by systematic review / meta-analysis (13.9%). The most prevalent article topics were treatment (39.6%), rehabilitation and return to play (11.9%), epidemiology (11.9%), and biomechanics (11.9%). Of the top 100 articles, 25.0% were American, 45.0% were European, and 30.0% were published outside of the United States or Europe. AAS had a strong correlation to Twitter mentions (r = 0.81), a moderate correlation to Google Scholar mentions (r = 0.52), and a weak correlation to Facebook mentions (r = 0.49). Conclusion: This study used AAS to characterize the 100 most impactful Achilles articles in online media. As access to research continues to move away from the conventional printed format, it is critically important to understand how orthopaedic information is disseminated online. Our findings suggest that alternative metrics broaden the definition of article impact beyond what is provided by traditional citation-based metrics and should be considered as a supplemental means of assessing the overall impact of published scientific literature.

Background: Rotator cuff repair using graft augmentation for large-to-massive, full-thickness rotator cuff tears has been reported to have improved clinical outcomes relative to other therapeutic interventions. Although an effective and promising technique, graft augmentation can be technically challenging, particularly with deployment and positioning of the graft. CuffMend is a user-friendly rotator cuff augmentation technique that combines a decellularized dermal allograft with a novel graft deployment device, tendon anchor, and set of lateral knotless, tension-able anchors to allow for a reliable and stable implantation of the graft. Indications: Graft augmentation using CuffMend is indicated for tears with a high risk of retear or incomplete healing. This includes revision tears, poor-quality degenerative tissue, and massive full-thickness rotator cuff tears. Severe glenohumeral osteoarthritic change is the primary contraindication to this technique. Technique Description: The rotator cuff is reduced medially and secured using 3 anchors with sutures passed in a mattress configuration. Graft augmentation is then performed using the CuffMend graft spreader that facilitates placement and attachment of the graft onto the repaired tendon. Results: The patient is recovering as expected and started physical therapy 1 month postoperatively. He is able to perform light activities of daily living and reach the top of his head for daily self-care without pain. He will begin strengthening at 12 weeks postoperatively. Discussion/Conclusion: CuffMend is a new graft augmentation technique that uses a novel graft deployment device for consistent and precise deployment of a decellularized dermal allograft. This facilitates an expedited repair using the graft augmentation technique for optimal clinical and biomechanical outcomes in patients with rotator cuff tears with high risk of retear.

Purpose: To evaluate and compare the Patient-Reported Outcomes Measurement Information System (PROMIS) Physical Function (PF) computerized adaptive test and PROMIS Pain Interference (PI) instruments versus legacy patient-reported outcome measures (PROMs) in patients undergoing hip arthroscopy for femoroacetabular impingement syndrome at 1-year follow-up. Methods: Patients undergoing primary hip arthroscopy for the treatment of femoroacetabular impingement syndrome without concomitant procedures performed by a single surgeon between August 2018 and January 2019 were identified. The PROMIS PF score, PROMIS PI score, Hip Outcome Score–Activities of Daily Living Subscale (HOS-ADL), Hip Outcome Score–Sports Subscale (HOS-SS), modified Harris Hip Score (mHHS), International Hip Outcome Tool 12 (iHOT-12), and visual analog scale (VAS) pain score were obtained preoperatively and at 6 months and 1 year postoperatively. Bivariate correlation analyses between PROMIS and legacy PROMs were performed. The floor and ceiling effects, responsiveness, and relative efficiency (RE) of each PROM were calculated. Results: This study included 124 patients with an average age of 32.7 ± 12.3 years. The PROMIS PF score showed very good correlations with the HOS-ADL (r = 0.73, P < .001) and mHHS (r = 0.70, P < .001) and good correlations with the HOS-SS (r = 0.62, P < .001), iHOT-12 score (r = 0.62, P < .001), and VAS pain score (r = –0.64, P < .001). The PROMIS PI score showed very good inverse correlations with the HOS-ADL (r = –0.72, P < .001) and mHHS (r = –0.79, P < .001) and good correlations with the HOS-SS (r = –0.64, P < .001), iHOT-12 score (r = –0.65, P < .001), and VAS pain score (r = 0.65, P < .001). No floor effect was observed for any measure. Ceiling effects were not observed in the PROMIS PROMs but were detected for the HOS-ADL (16.1%) and mHHS (19.3%). The effect size was large for all outcomes: iHOT-12 score, d = 1.77; HOS-ADL, d = 1.37; HOS-SS, d = 1.45; PROMIS PI score, d = 1.05; and PROMIS PF score, d = 1.01. The iHOT-12 score was more responsive than the PROMIS PI score (RE = 1.69), PROMIS PF score (RE = 1.75), HOS-ADL (RE = 1.29), and HOS-SS (RE = 1.22). Conclusions: At 1-year follow-up, PROMIS PROMs displayed very good correlations with legacy PROMs. However, PROMIS PROMs had lower effect sizes and were not as responsive as legacy PROMs. Level of Evidence: Level II, development of diagnostic criteria on the basis of consecutive patients.

Background: Medial patellar instability is rare but may occur after lateral retinacular release. The lateral patellofemoral ligament is a significant stabilizer for medial and lateral patellar displacement, and soft tissue–based reconstruction may restore its stabilizing effect. Indications: Lateral patellofemoral ligament reconstruction (LPFLR) is indicated for patellar instability failing exhaustive nonoperative measures. Technique Description: Diagnostic arthroscopy for medial patellofemoral ligament (MPFL) status, patellar tracking, chondral status, and postoperative changes of lateral retinacular release is conducted. A 5-cm incision is made between the midpoint of the patella and the lateral epicondyle, or a single midline incision can be used if performing a concomitant MPFL reconstruction. Dissection is carried down to the lateral retinaculum and capsule, and two 1-cm incisions are made at the distal and proximal patella through the patellar and quadriceps tendon, respectively. The lateral epicondyle is then directly palpated, and two 1-cm incisions are made directly anterior and posterior to the epicondyle into the iliotibial band in full extension. A hamstring allograft is then shuttled in a V-shaped configuration from the quadriceps tendon through the iliotibial band and finally to the patellar tendon. Excess graft is trimmed and doubled over proximally and distally to replicate the normal (or contralateral) 0° to 15° of patellar eversion. Trial resting length and tension is set with temporary sutures through the proximal and distal doubled graft in full extension. The knee is taken through full range of motion to ensure there is no graft tightening in flexion or loosening in full extension. Eversion and patellar translation are then examined. Heavy nonabsorbable sutures are used to set length of the doubled allograft proximally and distally. Results: Case series of LPFLR have demonstrated significant improvements in patients’ pain, Lysholm score and Knee Injury and Osteoarthritis Outcome Score, and kinesiophobia with no instability events and normal range of motion at short-term follow-up. Discussion/Conclusion: Medial patellar instability can occur following lateral retinacular release and patients experiencing instability despite nonoperative treatment are likely to benefit from a soft tissue–based LPFLR. Although other nongraft-based techniques may confer similar outcomes, the authors find it more reliable to perform a lateral reconstruction as described.

Background: Posterior instability is less common than anterior instability but can be seen in contact athletes and posttraumatically. Distal tibial allograft reconstruction for glenoid bone loss was first described by Provencher and colleagues in 2009 and an arthroscopic technique for posterior glenoid reconstruction using a distal tibial allograft was later described by Gupta et al in 2013. Indications: The primary indications for posterior distal tibial allograft include the failure of conservative management, recurrent instability after an arthroscopic stabilization, or glenoid bone loss > 20% to 25%. Technique Description: The patient is positioned in lateral decubitus, and examination under anesthesia is performed. Following arthroscopic evaluation, an incision is made medial to the posterolateral aspect of the acromion at the glenohumeral joint level. Electrocautery is carried to the deltoid, which is split in line with its fibers. A split between the infraspinatus and teres minor is performed. Vertical capsulotomy is performed, and deep retractors are placed. Attention is turned to the back table for graft preparation. The graft is measured, marked on the lateral aspect of the articular surface, and cut accordingly. Two 3.5-mm holes are drilled 1 cm apart, and the graft is thoroughly irrigated before being placed into the wound. A 2.5-mm drill is used in the 3.5-mm holes, and two 3.5-mm solid fully threaded screws are placed under power and tightened by hand. The wound is closed in the traditional fashion. Results: Graft nonunion and/or resorption are the primary concerns following posterior distal tibial allograft. Amar et al found no cases of nonunion or partial unions on 6-month computerized tomography (CT) scan, most patients having no or

Summary: Current conventional detection of SARS-CoV-2 involves collection of a patient’s sample with a nasopharyngeal swab, storage of the swab during transport in a viral transport medium, extraction of RNA, and quantitative reverse transcription PCR (RT-qPCR). We developed a simplified preparation method using a chelating resin, Chelex, which obviates RNA extraction during viral testing. Direct detection RT-qPCR and digital droplet PCR were compared to the current conventional method with RNA extraction for simulated samples and patient specimens. The heat treatment in the presence of Chelex markedly improved detection sensitivity as compared to heat alone, and lack of RNA extraction shortens the overall diagnostic workflow. Furthermore, the initial sample heating step inactivates SARS-CoV-2 infectivity, thus improving workflow safety. This fast RNA preparation and detection method is versatile for a variety of samples, safe for testing personnel, and suitable for standard clinical collection and testing on high-throughput platforms.

PurposeTo develop a novel method to quantify the amount of fibrosis in the salivary gland and to investigate the relationship between fibrosis and specific symptoms associated with Sjögren’s syndrome (SS) using this method.Materials and MethodsParaffin-embedded labial salivary gland (LSG) slides from 20 female SS patients and their clinical and LSG pathology data were obtained from the Sjögren’s International Collaborative Clinical Alliance. Relative interstitial fibrosis area (RIFA) in Masson’s trichrome-stained LSG sections was quantified from digitally scanned slides and used for correlation analysis. Gene expression levels were assessed by microarray analysis. Core promoter accessibility for RIFA-correlated genes was determined using DNase I hypersensitive sites sequencing analysis.ResultsRIFA was significantly correlated with unstimulated whole saliva flow rate in SS patients. Sixteen genes were significantly and positively correlated with RIFA. In a separate analysis, a group of differentially expressed genes was identified by comparing severe and moderate fibrosis groups. This combined set of genes was distinct from differentially expressed genes identified in lung epithelium from idiopathic pulmonary fibrosis patients compared with controls. Single-cell RNA sequencing analysis of salivary glands suggested most of the RIFA-correlated genes are expressed by fibroblasts in the gland and are in a permissive chromatin state.ConclusionRIFA quantification is a novel method for assessing interstitial fibrosis and the impact of fibrosis on SS symptoms. Loss of gland function may be associated with salivary gland fibrosis, which is likely to be driven by a unique set of genes that are mainly expressed by fibroblasts.

Greater trochanteric pain syndrome is a common cause of lateral hip pain, encompassing a spectrum of disorders, including trochanteric bursitis, abductor tendon pathology, and external coxa saltans. Greater trochanteric pain syndrome is primarily a clinical diagnosis, and careful clinical examination is essential for accurate diagnosis and treatment. A thorough history and physical exam may be used to help differentiate greater trochanteric pain syndrome from other common causes of hip pain, including osteoarthritis, femoroacetabular impingement, and lumbar stenosis. Although not required for diagnosis, plain radiographs and magnetic resonance imaging may be useful to exclude alternative pathologies or guide treatment of greater trochanteric pain syndrome. The majority of patients with greater trochanteric pain syndrome respond well to conservative management, including physical therapy, non-steroidal anti-inflammatory drugs, and corticosteroid injections. Operative management is typically indicated in patients with chronic symptoms refractory to conservative therapy. A wide range of surgical options, both open and endoscopic, are available and should be guided by the specific etiology of pain. The purpose of this review is to highlight pertinent clinical and radiographic features used in the diagnosis and management of greater trochanteric pain syndrome. In addition, treatment indications, techniques, and outcomes are described.

While great progress has been made in the understanding of neurological illnesses, the pathologies, and etiologies that give rise to these diseases still remain an enigma, thus, also making treatments for them more challenging. For effective and individualized treatment, it is beneficial to identify the underlying mechanisms that govern the associated cognitive and behavioral processes that go awry in neurological disorders. Parvalbumin fast-spiking interneurons (Pv-FSI) are GABAergic cells that are only a small fraction of the brain's neuronal network, but manifest unique cellular and molecular properties that drastically influence the downstream effects on signaling and ultimately change cognitive behaviors. Proper brain functioning relies heavily on neuronal communication which Pv-FSI regulates, excitatory-inhibitory balances and GABAergic disinhibition between circuitries. This review highlights the depth of Pv-FSI involvement in the cortex, hippocampus, and striatum, as it pertains to expression, neurotransmission, role in neurological disorders, and dysfunction, as well as cognitive behavior and reward-seeking. Recent research has indicated that Pv-FSI play pivotal roles in the molecular pathophysiology and cognitive-behavioral deficits that are core features of many psychiatric disorders, such as schizophrenia, autism spectrum disorders, Alzheimer's disease, and drug addiction. This suggests that Pv-FSI could be viable targets for treatment of these disorders and thus calls for further examination of the undeniable impact Pv-FSI have on the brain and cognitive behavior.

Here, the authors explore the potential of the 16S gene for discriminating bacterial taxa and show that full-length sequencing combined with appropriate clustering of intragenomic sequence variation can provide accurate representation of bacterial species in microbiome datasets.

Background: Sjögren's syndrome (SS) is one of the most common autoimmune disorders leading to exocrine gland dysfunction. Both immune-dependent processes – like Type I Interferon (IFN) signaling and immune-independent processes – such as calcium signaling in epithelial cells – contribute to disease pathophysiology. However, a mechanistic link between these processes has not been demonstrated. Methods: Primary human salivary gland cells were used to evaluate the differential expression of miRNAs with smRNA-seq in primary epithelial cells culture and digital PCR was conducted in SS human salivary glands (SG) biopsies to verify the results. With siRNA screening and pull-down assays to establish the role of miRNA in IFN activation. Findings: Activation of IFN-β by miR-1248 is through the direct association with both RIG-I and AGO2. Further functional studies establish a unique dual functional role of miR-1248 in phSG cells: i) activation of the RIG-I pathway by acting as ligand of this sensor leading to IFN production and ii) regulation of the expression of mRNAs through the canonical microRNA function. Importantly, ITPR3, a key component of calcium signaling in epithelial cells, that has previously shown to be downregulated in SS SG, was directly targeted and downregulated by miR-1248, inducing the same functional calcium signaling changes as observed in SS SGs. Interpretation: Identification of the first endogenous mammalian microRNA that binds to RIG-I inducing IFN production but also demonstrate a novel pathophysiological underlying mechanism in which miR-1248 overexpression links two major pathways associated with SS, namely activation of IFN production with modulation of calcium signaling. Together, these findings suggest a unifying hypothesis for the immune-independent and -dependent processes contributing to the pathogenesis of SS. Fund: This research was supported by the Intramural Research Program of the National Institutes of Health (NIH), National Institute of Dental and Craniofacial Research (NIDCR). Keywords: Sjögren's syndrome, miR-1248, Interferon, RIG-I and ITPR3

Hemorrhage associated with anticoagulation therapy is a feared and relatively common complication. Few cases have been documented of spontaneous pancreatic hemorrhage related to anticoagulation, and fewer yet of ruptured pancreatic pseudocyst secondary to anticoagulation-related bleeding. We describe the case of a 74-year-old female with massive intra- and extraperitoneal hemorrhage secondary to an anticoagulation-related ruptured hemorrhagic pancreatic pseudocyst that was treated successfully with coil embolization. An inferior vena cava filter was placed at the same time as the embolization procedure, as she had a contraindication for anticoagulation in the setting of submassive pulmonary emboli and lower extremity deep venous thrombosis. Keywords: Embolization, Anticoagulation, Hemorrhage, Pancreas, Pseudocyst

With an increased interest in youth sports, the burden of overhead throwing elbow injuries accompanying early single-sport focus has steadily risen. During the overhead throwing motion, valgus torque can reach and surpass Newton meters (N m) during the late cocking and early acceleration phases, which exceeds the tensile strength (22.7–33 N m) of the ulnar collateral ligament. While the ulnar collateral ligament serves as the primary valgus stabilizer between and degrees of elbow flexion, other structures about the elbow must contribute to stability during throwing. Depending on an athlete’s stage of skeletal maturity, certain patterns of injury are observed with mechanical failures resulting from increased medial laxity, lateral-sided compression, and posterior extension shearing forces. Together, these injury patterns represent a wide range of conditions that arise from valgus extension overload. The purpose of this article is to review common pathologies observed in the adolescent overhead throwing athlete in the context of functional anatomy, osseous development, and throwing mechanics. Operative and non-operative management and their associated outcomes will be discussed for these injuries.