Hamstring Injuries: A Paradigm for Return to Play.

Muscle injuries are the most common sports-related injuries, with hamstring involvement most common in professional athletes. These injuries can lead to significant time lost from play and have a high risk of reinjury. We review the anatomy, mechanisms of injury, diagnostic imaging modalities, and treatment techniques for hamstring injuries. We also present the latest evidence related to return to play (RTP) after hamstring injuries, including a review of articles targeted to RTP in European soccer (Union of European Football Associations), American football (National Football League), and other professional sports. Review of imaging findings in hamstring injury, grading systems for injuries, considerations for RTP, as well as advances in injury prevention, are discussed.

Deep Learning-Assisted Identification of Femoroacetabular Impingement (FAI) on Routine Pelvic Radiographs.

To use a novel deep learning system to localize the hip joints and detect findings of cam-type femoroacetabular impingement (FAI). A retrospective search of hip/pelvis radiographs obtained in patients to evaluate for FAI yielded 3050 total studies. Each hip was classified separately by the original interpreting radiologist in the following manner: 724 hips had severe cam-type FAI morphology, 962 moderate cam-type FAI morphology, 846 mild cam-type FAI morphology, and 518 hips were normal. The anteroposterior (AP) view from each study was anonymized and extracted. After localization of the hip joints by a novel convolutional neural network (CNN) based on the focal loss principle, a second CNN classified the images of the hip as cam positive, or no FAI. Accuracy was 74% for diagnosing normal vs. abnormal cam-type FAI morphology, with aggregate sensitivity and specificity of 0.821 and 0.669, respectively, at the chosen operating point. The aggregate AUC was 0.736. A deep learning system can be applied to detect FAI-related changes on single view pelvic radiographs. Deep learning is useful for quickly identifying and categorizing pathology on imaging, which may aid the interpreting radiologist.

Enhanced fungal specificity and in vivo therapeutic efficacy of a C-22-modified FK520 analog against C. neoformans.

IMPORTANCE: Fungal infections cause significant morbidity and mortality globally. The therapeutic armamentarium against these infections is limited, and the development of antifungal drugs has been hindered by the evolutionary conservation between fungi and the human host. With rising resistance to the current antifungal arsenal and an increasing at-risk population, there is an urgent need for the development of new antifungal compounds. The FK520 analogs described in this study display potent antifungal activity as a novel class of antifungals centered on modifying an existing orally active FDA-approved therapy. This research advances the development of much-needed newer antifungal treatment options with novel mechanisms of action.

A semi-quantitative, rapid, point of care SARS-CoV-2 serologic assay predicts neutralizing antibody levels.

The ongoing COVID-19 pandemic has caused millions of deaths and the continued emergence of new variants suggests continued circulation in the human population. In the current time of vaccine availability and new therapeutic development, including antibody-based therapies, many questions about long-term immunity and protection remain uncertain. Identification of protective antibodies in individuals is often done using highly specialized and challenging assays such as functional neutralizing assays, which are not available in the clinical setting. Therefore, there is a great need for the development of rapid, clinically available assays that correlate with neutralizing antibody assays to identify individuals who may benefit from additional vaccination or specific COVID-19 therapies. In this report, we apply a novel semi-quantitative method to an established lateral flow assay (sqLFA) and analyze its ability to detect the presence functional neutralizing antibodies from the serum of COVID-19 recovered individuals. We found that the sqLFA has a strong positive correlation with neutralizing antibody levels. At lower assay cutoffs, the sqLFA is a highly sensitive assay to identify the presence of a range of neutralizing antibody levels. At higher cutoffs, it can detect higher levels of neutralizing antibody with high specificity. This sqLFA can be used both as a screening tool to identify individuals with any level of neutralizing antibody to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), or as a more specific tool to identify those with high neutralizing antibody levels who may not benefit from antibody-based therapies or further vaccination.

Enhanced fungal specificity and in vivo therapeutic efficacy of a C-22 modified FK520 analog against C. neoformans.

Fungal infections are of mounting global concern, and the current limited treatment arsenal poses challenges when treating such infections. In particular, infections by Cryptococcus neoformans are associated with high mortality, emphasizing the need for novel therapeutic options. Calcineurin is a protein phosphatase that mediates fungal stress responses, and calcineurin inhibition by the natural product FK506 blocks C. neoformans growth at 37°C. Calcineurin is also required for pathogenesis. However, because calcineurin is conserved in humans, and inhibition with FK506 results in immunosuppression, the use of FK506 as an anti-infective agent is precluded. We previously elucidated the structures of multiple fungal calcineurin-FK506-FKBP12 complexes and implicated the C-22 position on FK506 as a key point for differential modification of ligand inhibition of the mammalian versus fungal target proteins. Through in vitro antifungal and immunosuppressive testing of FK520 (a natural analog of FK506) derivatives, we identified JH-FK-08 as a lead candidate for further antifungal development. JH-FK-08 exhibited significantly reduced immunosuppressive activity and both reduced fungal burden and prolonged survival of infected animals. JH-FK-08 exhibited additive activity in combination with fluconazole in vivo . These findings further advance calcineurin inhibition as an antifungal therapeutic approach. Importance: Fungal infections cause significant morbidity and mortality globally. The therapeutic armamentarium against these infections is limited and development of antifungal drugs has been hindered by the evolutionary conservation between fungi and the human host. With rising resistance to the current antifungal arsenal and an increasing at-risk population, there is an urgent need for the development of new antifungal compounds. The FK520 analogs described in this study display potent antifungal activity as a novel class of antifungals centered on modifying an existing orally-active FDA approved therapy. This research advances the development of much needed newer antifungal treatment options with novel mechanisms of action.

Drug Target Elucidation Through Isolation and Analysis of Drug-Resistant Mutants in Cryptococcus neoformans.

Drug target identification is an essential component to antifungal drug development. Many methods, including large chemical library screening, natural product screening, and drug repurposing efforts, can identify compounds with favorable in vitro antifungal activity. However, these approaches will often identify compounds with no known mechanism of action. Herein, we describe a method utilizing the human fungal pathogen Cryptococcus neoformans to identify antifungal drug targets through the isolation of spontaneous resistant mutants, antifungal testing, whole-genome sequencing, and variant analysis.

Development and performance of a point-of-care rapid antigen test for detection of SARS-COV-2 variants.

Background: SARS-CoV-2 antigen-based tests are well-calibrated to infectiousness and have a critical role to play in the COVID-19 public health response. We report the development and performance of a unique lateral flow immunoassay (LFA). Methods: Combinations of several monoclonal antibodies targeting multiple antigenic sites on the SARS-CoV-2 nucleocapsid protein (NP) were isolated, evaluated, and chosen for the development of a LFA termed CoV-SCAN (BioMedomics, Inc.). Clinical point-of-care studies in symptomatic and asymptomatic individuals were conducted to evaluate positive predictive agreement (PPA) and negative predictive agreement (NPA) with RT-PCR as comparator. Results: In laboratory testing, CoV-SCAN detected 14 recombinant N-proteins of SARS-CoV-2 variants with sensitivity in the range of 0.2-3.2 ng/mL, and 10 authentic SARS-CoV-2 variants with sensitivity in the range of 1.6-12.5 TCID50/swab. No cross reactivity was observed with other human coronaviruses or other respiratory pathogens. In clinical point-of-care testing on 148 individuals over age 2 with symptoms of ≤5 days, PPA was 87.2% (CI 95: 78.3-94.8%) and NPA was 100% (CI 95: 94.2-100%). In another 884 asymptomatic individuals, PPA was 85.7% (CI 95: 42.1-99.6%) and 99.7% (99.0-99.9%). Overall, CoV-SCAN detected over 97.2% of specimens with CT values <30 and 93.8% of nasal swab specimens with the Omicron variant, even within the first 2 days after symptom onset. Conclusions: The unique construction of CoV-SCAN using two pairs of monoclonal antibodies has resulted in a test with high performance that remains durable across multiple variants in both laboratory and clinical evaluations. CoV-SCAN should identify almost all individuals harboring infectious SARS-CoV-2. Summary: Unique construction of a point-of-care rapid antigen test using two pairs of monoclonal antibodies has led to good performance that remained durable across multiple variants in laboratory and clinical evaluations. Test should identify almost all individuals harboring infectious SARS-CoV-2.

Structure-Guided Synthesis of FK506 and FK520 Analogs with Increased Selectivity Exhibit In Vivo Therapeutic Efficacy against Cryptococcus.

Calcineurin is an essential virulence factor that is conserved across human fungal pathogens, including Cryptococcus neoformans, Aspergillus fumigatus, and Candida albicans. Although an excellent target for antifungal drug development, the serine-threonine phosphatase activity of calcineurin is conserved in mammals, and inhibition of this activity results in immunosuppression. FK506 (tacrolimus) is a naturally produced macrocyclic compound that inhibits calcineurin by binding to the immunophilin FKBP12. Previously, our fungal calcineurin-FK506-FKBP12 structure-based approaches identified a nonconserved region of FKBP12 that can be exploited for fungus-specific targeting. These studies led to the design of an FK506 analog, APX879, modified at the C-22 position, which was less immunosuppressive yet maintained antifungal activity. We now report high-resolution protein crystal structures of fungal FKBP12 and a human truncated calcineurin-FKBP12 bound to a natural FK506 analog, FK520 (ascomycin). Based on information from these structures and the success of APX879, we synthesized and screened a novel panel of C-22-modified compounds derived from both FK506 and FK520. One compound, JH-FK-05, demonstrates broad-spectrum antifungal activity in vitro and is nonimmunosuppressive in vivo. In murine models of pulmonary and disseminated C. neoformans infection, JH-FK-05 treatment significantly reduced fungal burden and extended animal survival alone and in combination with fluconazole. Furthermore, molecular dynamic simulations performed with JH-FK-05 binding to fungal and human FKBP12 identified additional residues outside the C-22 and C-21 positions that could be modified to generate novel FK506 analogs with improved antifungal activity. IMPORTANCE Due to rising rates of antifungal drug resistance and a limited armamentarium of antifungal treatments, there is a paramount need for novel antifungal drugs to treat systemic fungal infections. Calcineurin has been established as an essential and conserved virulence factor in several fungi, making it an attractive antifungal target. However, due to the immunosuppressive action of calcineurin inhibitors, they have not been successfully utilized clinically for antifungal treatment in humans. Recent availability of crystal structures of fungal calcineurin-bound inhibitor complexes has enabled the structure-guided design of FK506 analogs and led to a breakthrough in the development of a compound with increased fungal specificity. The development of a calcineurin inhibitor with reduced immunosuppressive activity and maintained therapeutic antifungal activity would add a significant tool to the treatment options for these invasive fungal infections with exceedingly high rates of mortality.

Long-term Safety and Efficacy of the Anti-MAdCAM-1 Monoclonal Antibody Ontamalimab [SHP647] for the Treatment of Ulcerative Colitis: The Open-label Study TURANDOT II.

BACKGROUND AND AIMS: Ontamalimab, a fully-human monoclonal antibody targeting MAdCAM-1, induced remission in patients with moderate-to-severe ulcerative colitis [UC] in the TURANDOT study. We aimed to assess long-term safety, tolerability, and efficacy of ontamalimab in TURANDOT II. METHODS: TURANDOT II was a phase 2, multicentre, open-label [OL] study in patients with moderate-to-severe UC who completed TURANDOT on placebo or ontamalimab (NCT01771809). Patients were randomised to 75 mg or 225 mg ontamalimab every 4 weeks for 72 weeks [OL1]. The dosage could be increased to 225 mg from Week 8 at the investigator's discretion. All patients then received 75 mg every 4 weeks for 72 weeks [OL2], followed by 6-month safety follow-up. The primary objective was safety, measured by adverse events [AEs], serious AEs [SAEs], and AEs leading to withdrawal. Mucosal healing [MH; centrally read endoscopy] was assessed. RESULTS: Of 330 patients, 180 completed OL1; 94 escalated to 225 mg; 127 completed OL2. Overall, 36.1% experienced drug-related AEs. The most common SAE [10.0%] was worsening/ongoing UC; 5.5% of patients had serious infections, the most common being gastroenteritis [0.9%]. One death and four cancers [all unrelated to ontamalimab] occurred. No PML [progressive multifocal leukoencephalopathy]/lymphoproliferative disorders occurred. Geometric mean high-sensitivity C-reactive protein [hsCRP] and faecal calprotectin decreased across OL1 in both dose groups. The proportion of patients assigned to placebo in TURANDOT achieving MH increased from 8.8% [6/68] at baseline to 35.3% at Week 16 [24/68; non-responder imputation]. The corresponding increase in the ontamalimab group was from 23.3% [61/262] to 26.7% [70/262]. CONCLUSIONS: Ontamalimab was well tolerated up to 144 weeks in patients with moderate-to-severe UC, with good safety and efficacy.

Fungal pathogens.

Fungi are key components in global biogeochemical cycles, play important roles in manufacturing industries and biomedical research, and influence humans through their impact on global health, agriculture, and biodiversity. Fungi have been isolated from almost every environmental niche across the planet, including from air, soil, fresh water, and the oceans. Although the vast majority of fungi do not exhibit pathogenic traits, some species cause infections in humans ranging from superficial to life-threatening. Moreover, some fungal species are plant pathogens and have devastating impacts on agriculture. In this primer, we aim to provide a broad picture of what makes fungal pathogens unique, as well as the challenges of combating fungal pathogens.

The Search for Optimal Oxygen Saturation Targets in Critically Ill Patients: Observational Data From Large ICU Databases.

BACKGROUND: Although low oxygen saturations are generally regarded as deleterious, recent studies in ICU patients have shown that a liberal oxygen strategy increases mortality. However, the optimal oxygen saturation target remains unclear. The goal of this study was to determine the optimal range by using real-world data. METHODS: Replicate retrospective analyses were conducted of two electronic medical record databases: the eICU Collaborative Research Database (eICU-CRD) and the Medical Information Mart for Intensive Care III database (MIMIC). Only patients with at least 48 h of oxygen therapy were included. Nonlinear regression was used to analyze the association between median pulse oximetry-derived oxygen saturation (Spo2) and hospital mortality. We derived an optimal range of Spo2 and analyzed the association between the percentage of time within the optimal range of Spo2 and hospital mortality. All models adjusted for age, BMI, sex, and Sequential Organ Failure Assessment score. Subgroup analyses included ICU types, main diagnosis, and comorbidities. RESULTS: The analysis identified 26,723 patients from eICU-CRD and 8,564 patients from MIMIC. The optimal range of Spo2 was 94% to 98% in both databases. The percentage of time patients were within the optimal range of Spo2 was associated with decreased hospital mortality (OR of 80% vs 40% of the measurements within the optimal range, 0.42 [95% CI, 0.40-0.43] for eICU-CRD and 0.53 [95% CI, 0.50-0.55] for MIMIC). This association was consistent across subgroup analyses. CONCLUSIONS: The optimal range of Spo2 was 94% to 98% and should inform future trials of oxygen therapy.

Harnessing calcineurin-FK506-FKBP12 crystal structures from invasive fungal pathogens to develop antifungal agents.

Calcineurin is important for fungal virulence and a potential antifungal target, but compounds targeting calcineurin, such as FK506, are immunosuppressive. Here we report the crystal structures of calcineurin catalytic (CnA) and regulatory (CnB) subunits complexed with FK506 and the FK506-binding protein (FKBP12) from human fungal pathogens (Aspergillus fumigatus, Candida albicans, Cryptococcus neoformans and Coccidioides immitis). Fungal calcineurin complexes are similar to the mammalian complex, but comparison of fungal and human FKBP12 (hFKBP12) reveals conformational differences in the 40s and 80s loops. NMR analysis, molecular dynamic simulations, and mutations of the A. fumigatus CnA/CnB-FK506-FKBP12-complex identify a Phe88 residue, not conserved in hFKBP12, as critical for binding and inhibition of fungal calcineurin. These differences enable us to develop a less immunosuppressive FK506 analog, APX879, with an acetohydrazine substitution of the C22-carbonyl of FK506. APX879 exhibits reduced immunosuppressive activity and retains broad-spectrum antifungal activity and efficacy in a murine model of invasive fungal infection.

The Effect of Turf Toe Injuries on Player Performance in the National Football League.

Background: "Turf toe" results from hyperdorsiflexion of the first metatarsophalangeal joint, injuring the plantar capsuloligamentous complex. We hypothesized that National Football League (NFL) player performance following turf toe injury would decrease in comparison to controls at the same position. Methods: Demographics, return to play, and season performance data on players sustaining turf toe injuries in the NFL from 2010-2015 were collected. An Offensive Power Rating (OPR=[total yards/10]+[total touchdowns x6]) or Defensive Power Rating (DPR=total tackles+[total sacks x2]+[total interceptions x2]) was calculated for each player. Control data were collected for NFL players in 2013 with no history of turf toe injury. Statistical analysis was performed using Wilcoxon Rank Sum tests. Results: Twenty-four injured players and 436 controls were included. Nineteen players returned to play within the regular season of injury (mean 36.7 ± 28.9 days). Seventeen players were removed from team injury reports for turf toe within the regular season (mean 42.6 ± 26.2 days). Three players required season-ending surgery. Comparison of 1-year post- versus pre-injury revealed an insignificant median OPR difference (-18.9 IQR -43.4 to 10.3 vs. control -12.2 IQR -46.2 to 47.7, p = 0.328) and median DPR difference (-1.0 IQR -26.0 to 17.0 vs. control 2.0 IQR -15.0 to 18.0, p = NA). Comparison of 2-year data revealed no significant median OPR difference (-32.6 IQR -122.2 to 1.0 vs. control -20.7 IQR -72.6 to 44.7, p = 0.327) and median DPR difference (-5.0 IQR -19.0 to 6.0 vs. control -4.5 IQR -22.0 to 12.5, p= NA). Conclusions: Turf toe results in significant loss of playing time. Despite the long recovery period, NFL players have similar performance following injury compared to controls. The effect of turf toe injuries on performance is variable.Level of evidence: IV.

A comparison of narcotic usage and length of post-operative hospital stay in open versus minimally invasive lumbar interbody fusion with percutaneous pedicle screws.

BACKGROUND: Instrumented lumbar fusion can be accomplished through open or minimally invasive techniques. The focus of this study was to compare perioperative narcotic usage and length of hospital stay between patients undergoing open versus minimally invasive spinal surgery (MISS). METHODS: A retrospective chart review was performed on 110 patients who underwent instrumented lumbar fusion over 2 years at our institution. These patients were divided into two groups: those that received open transforaminal interbody fusion (n=69), and those whose surgeries were performed minimally invasively with lateral lumbar transpsoas interbody fusion (LLIF) and percutaneous pedicle screws (n=41). Narcotic usage was recorded for both groups intra-operatively and post-operatively throughout their hospital stay. These values were standardized using an equianalgesia chart. RESULTS: Average narcotic usage post-operatively was significantly lower for the LLIF group relative to those who underwent open lumbar fusion (278.48 vs. 442.06 mg, P=0.03). The average length of post-operative hospital stay was significantly shorter for patients who underwent LLIF compared to those who had an open procedure (4.10 vs. 6.19 days, P=0.02). CONCLUSIONS: Patients who underwent minimally invasive surgery (MIS) LLIF had decreased overall use of opioids in the perioperative period and shorter hospital stays when compared to patients who underwent the open transforaminal interbody fusion approach. These findings support pre-existing literature in favor of LLIF MISS with regards to the above stated outcome measures. The long-term benefits of MISS with regards to narcotic usage in spine patients are not yet known.

Evolutionarily Conserved and Divergent Roles of Unfolded Protein Response (UPR) in the Pathogenic Cryptococcus Species Complex.

The unfolded protein response (UPR) pathway, consisting of the evolutionarily conserved Ire1 kinase/endonuclease and the bZIP transcription factor Hxl1, is critical for the pathogenicity of Cryptococcus neoformans; however, its role remains unknown in other pathogenic Cryptococcus species. Here, we investigated the role of the UPR pathway in C. deuterogattii, which causes pneumonia and systemic cryptococcosis, even in immunocompetent individuals. In response to ER stress, C. deuterogattii Ire1 triggers unconventional splicing of HXL1 to induce the expression of UPR target genes such as KAR2, DER1, ALG7, and ERG29. Furthermore, C. deuterogattii Ire1 is required for growth at mammalian body temperature, similar to C. neoformans Ire1. However, deletion of HXL1 does not significantly affect the growth of C. deuterogattii at 37 °C, which is in contrast to the indispensable role of HXL1 in the growth of C. neoformans at 37 °C. Nevertheless, both C. deuterogattii ire1Δ and hxl1Δ mutants are avirulent in a murine model of systemic cryptococcosis, suggesting that a non-thermotolerance phenotypic trait also contributes to the role of the UPR pathway in the virulence of pathogenic Cryptococcus species. In conclusion, the UPR pathway plays redundant and distinct roles in the virulence of members of the pathogenic Cryptococcus species complex.

A complex obstetric case.

A 33-year-old primigravida presented with severe sepsis, severe pre-eclampsia, peripartum cardiomyopathy and the haemolysis, elevated liver enzymes and low platelets syndrome manifesting over the course of 24 h causing a diagnostic conundrum and a difficult sequence of physiological problems to overcome. We describe a previously unreported sequence of events involving a pre-eclamptic, septic parturient to improve anaesthetic and intensive care physician awareness of confounding factors that complicate assessment and management of these patients.

Metabolic network rewiring of propionate flux compensates vitamin B12 deficiency in C. elegans.

Metabolic network rewiring is the rerouting of metabolism through the use of alternate enzymes to adjust pathway flux and accomplish specific anabolic or catabolic objectives. Here, we report the first characterization of two parallel pathways for the breakdown of the short chain fatty acid propionate in Caenorhabditis elegans. Using genetic interaction mapping, gene co-expression analysis, pathway intermediate quantification and carbon tracing, we uncover a vitamin B12-independent propionate breakdown shunt that is transcriptionally activated on vitamin B12 deficient diets, or under genetic conditions mimicking the human diseases propionic- and methylmalonic acidemia, in which the canonical B12-dependent propionate breakdown pathway is blocked. Our study presents the first example of transcriptional vitamin-directed metabolic network rewiring to promote survival under vitamin deficiency. The ability to reroute propionate breakdown according to B12 availability may provide C. elegans with metabolic plasticity and thus a selective advantage on different diets in the wild.

Analyzing health insurance claims on different timescales to predict days in hospital.

Health insurers maintain large databases containing information on medical services utilized by claimants, often spanning several healthcare services and providers. Proper use of these databases could facilitate better clinical and administrative decisions. In these data sets, there exists many unequally spaced events, such as hospital visits. However, data mining of temporal data and point processes is still a developing research area and extracting useful information from such data series is a challenging task. In this paper, we developed a time series data mining approach to predict the number of days in hospital in the coming year for individuals from a general insured population based on their insurance claim data. In the proposed method, the data were windowed at four different timescales (bi-monthly, quarterly, half-yearly and yearly) to construct regularly spaced time series features extracted from such events, resulting in four associated prediction models. A comparison of these models indicates models using a half-yearly windowing scheme delivers the best performance on all three populations (the whole population, a senior sub-population and a non-senior sub-population). The superiority of the half-yearly model was found to be particularly pronounced in the senior sub-population. A bagged decision tree approach was able to predict 'no hospitalization' versus 'at least one day in hospital' with a Matthews correlation coefficient (MCC) of 0.426. This was significantly better than the corresponding yearly model, which achieved 0.375 for this group of customers. Further reducing the length of the analysis windows to three or two months did not produce further improvements.

High-glucose diets have sex-specific effects on aging in C. elegans: toxic to hermaphrodites but beneficial to males.

Diet and sex are important determinants of lifespan. In humans, high sugar diets, obesity, and type 2 diabetes correlate with decreased lifespan, and females generally live longer than males. The nematode Caenorhabditis elegans is a classical model for aging studies, and has also proven useful for characterizing the response to high-glucose diets. However, studies on male animals are lacking. We found a surprising dichotomy: glucose regulates lifespan and aging in a sex-specific manner, with beneficial effects on males compared to toxic effects on hermaphrodites. High-glucose diet resulted in greater mobility with age for males, along with a modest increase in median lifespan. In contrast, high-glucose diets decrease both lifespan and mobility for hermaphrodites. Understanding sex-specific responses to high-glucose diets will be important for determining which evolutionarily conserved glucose-responsive pathways that regulate aging are "universal" and which are likely to be cell-type or sex-specific.