Improved Trajectory Reconstruction for Markerless Pose Estimation.

Markerless pose estimation allows reconstructing human movement from multiple synchronized and calibrated views, and has the potential to make movement analysis easy and quick, including gait analysis. This could enable much more frequent and quantitative characterization of gait impairments, allowing better monitoring of outcomes and responses to interventions. However, the impact of different keypoint detectors and reconstruction algorithms on markerless pose estimation accuracy has not been thoroughly evaluated. We tested these algorithmic choices on data acquired from a multicamera system from a heterogeneous sample of 53 individuals in a rehabilitation hospital. We found that using a top-down keypoint detector and reconstructing trajectories with an implicit function enabled accurate, smooth, and anatomically plausible trajectories, with a noise in the step width estimates compared to a GaitRite walkway of only 9mm.

Machine Learning-Based Gait Mode Prediction for Hybrid Knee Prosthesis Control.

Recently, hybrid prosthetic knees, which can combine the advantages of passive and active prosthetic knees, have been proposed for individuals with a transfemoral amputation. Users could potentially take advantage of the passive knee mechanics during walking and the active power generation during stair ascent. One challenge in controlling the hybrid knees is accurate gait mode prediction for seamless transitions between passive and active modes. However, data imbalance between passive and active modes may impact the performance of a classifier. In this study, we used a dataset collected from nine individuals with a unilateral transfemoral amputation as they ambulated over level ground, inclines, and stairs. We evaluated several machine learning-based classifiers on the prediction of passive (level-ground walking, incline walking, descending stairs, and donning and doffing the prosthesis) and active mode (ascending stairs). In addition, we developed a generative adversarial network (GAN) to create synthetic data for improving classification performance. The results indicated that linear discriminant analysis and random forest might be the best classifiers regarding sensitivity to the active mode and overall accuracy, respectively. Further, we demonstrated that using the GAN-based synthetic data for training improves the sensitivity of classifiers.

Optimizing Trajectories and Inverse Kinematics for Biomechanical Analysis of Markerless Motion Capture Data.

Markerless motion capture using computer vision and human pose estimation (HPE) has the potential to expand access to precise movement analysis. This could greatly benefit rehabilitation by enabling more accurate tracking of outcomes and providing more sensitive tools for research. There are numerous steps between obtaining videos to extracting accurate biomechanical results and limited research to guide many critical design decisions in these pipelines. In this work, we analyze several of these steps including the algorithm used to detect keypoints and the keypoint set, the approach to reconstructing trajectories for biomechanical inverse kinematics and optimizing the IK process. Several features we find important are: 1) using a recent algorithm trained on many datasets that produces a dense set of biomechanically-motivated keypoints, 2) using an implicit representation to reconstruct smooth, anatomically constrained marker trajectories for IK, 3) iteratively optimizing the biomechanical model to match the dense markers, 4) appropriate regularization of the IK process. Our pipeline makes it easy to obtain accurate biomechanical estimates of movement in a rehabilitation hospital.

Signal transduction at GPCRs: Allosteric activation of the ERK MAPK by ß-arrestin.

ß-arrestins are multivalent adaptor proteins that bind active phosphorylated G protein-coupled receptors (GPCRs) to inhibit G protein signaling, mediate receptor internalization, and initiate alternative signaling events. ß-arrestins link agonist-stimulated GPCRs to downstream signaling partners, such as the c-Raf-MEK1-ERK1/2 cascade leading to ERK1/2 activation. ß-arrestins have been thought to transduce signals solely via passive scaffolding by facilitating the assembly of multiprotein signaling complexes. Recently, however, ß-arrestin 1 and 2 were shown to activate two downstream signaling effectors, c-Src and c-Raf, allosterically. Over the last two decades, ERK1/2 have been the most intensely studied signaling proteins scaffolded by ß-arrestins. Here, we demonstrate that ß-arrestins play an active role in allosterically modulating ERK kinase activity in vitro and within intact cells. Specifically, we show that ß-arrestins and their GPCR-mediated active states allosterically enhance ERK2 autophosphorylation and phosphorylation of a downstream ERK2 substrate, and we elucidate the mechanism by which ß-arrestins do so. Furthermore, we find that allosteric stimulation of dually phosphorylated ERK2 by active-state ß-arrestin 2 is more robust than by active-state ß-arrestin 1, highlighting differential capacities of ß-arrestin isoforms to regulate effector signaling pathways downstream of GPCRs. In summary, our study provides strong evidence for a new paradigm in which ß-arrestins function as active "catalytic" scaffolds to allosterically unlock the enzymatic activity of signaling components downstream of GPCR activation.

Relationship among DDR gene mutations, TMB and PD-L1 in solid tumour genomes identified using clinically actionable biomarker assays.

The DNA damage response (DDR) pathway regulates DNA repair and cell survival, and inactivating mutations in DDR genes can increase tumour mutational burden (TMB), a predictive biomarker of treatment benefit from anti-PD-1/PD-L1 immunotherapies. However, a better understanding of the relationship among specific DDR mutations, TMB and PD-L1 expression is needed to improve translational strategies. Here, we determined genomic alteration frequencies in selected DDR genes that are clinically actionable biomarkers and investigated their association with TMB and PD-L1 in bladder, colorectal, non-small cell lung, ovarian and prostate cancers using the FoundationInsights® web portal. Our results not only confirm known associations, such as mismatch repair and POLE gene mutations with high TMB, but also identify significant associations between mutations in the SWI/SNF chromatin remodelling genes ARID1A and SMARCA4 and high TMB in multiple tumour types. Mutations in the ATR gene were associated with high TMB in colorectal and prostate cancers; however, associations between individual DDR mutations and high PD-L1 expression were uncommon and tumour-type specific. Finally, we found that high TMB and high PD-L1 expression were poorly associated, emphasising their independence as predictive biomarkers for immune checkpoint inhibitor use.

Impact of HEART Score Decision Aid on Coronary Computed Tomography Angiography Utilization and Diagnostic Yield in the Emergency Department.

OBJECTIVE: Emergency physicians are challenged to efficiently and reliably risk stratify patients presenting with chest pain (CP) to optimize diagnostic testing and avoid unnecessary hospital admissions. The objective of our study was to evaluate the impact of a HEART score-based decision aid (HSDA) integrated in the electronic health record on coronary computed tomography angiography (CCTA) utilization and diagnostic yield in adult emergency department (ED) CP patients with suspected acute coronary syndrome. METHODS: We conducted a before and after study to determine whether implementation of a mandatory computerized HSDA would reduce CCTA utilization in ED CP patients and improve the diagnostic yield of obstructive coronary artery disease (CAD) (≥50%). We included all adult ED CP patients with suspected acute coronary syndrome during the first 6 months of 2018 (before) and 2020 (after) at a large academic center. CCTA utilization and obstructive CAD yield were compared in patients before and after implementing the HSDA using χ2 tests. Secondarily, we assessed the association of HEART scores and CCTA results. RESULTS: Of the 3095 CP patients during the before study period, 733 underwent CCTA. Of the 2692 CP patients during the after study period, 339 underwent CCTA. CCTA utilization before and after HSDA was 23.4% [95% confidence interval (95% CI), 22.2-25.2] and 12.6% (95% CI, 11.4-13.0), respectively; mean difference was 11.1% (95% CI, 0.9-13.0). Among 1072 patients undergoing CCTA, mean (SD) age and percent females before versus after HSDA were 54 (11) versus 56 (11) years and 50% versus 49%, respectively. We included 1014 patients (686 before and 328 after) for the yield analysis. Obstructive CAD was present in 15% (95% CI, 12.7-17.9) and 20.1% (95% CI, 16.1-24.7) before and after HSDA, respectively; mean difference was 4.9% (95% CI, 0.1-10.1). CONCLUSIONS: Implementation of a mandatory electronic health record HSDA aid reduced ED CCTA utilization by half and improved the diagnostic yield.

Mismatch repair deficiency, next-generation sequencing-based microsatellite instability, and tumor mutational burden as predictive biomarkers for immune checkpoint inhibitor effectiveness in frontline treatment of advanced stage endometrial cancer.

OBJECTIVE: Molecular profiling is developing to inform treatment in endometrial cancer. Using real world evidence, we sought to evaluate frontline immune checkpoint inhibitor vs chemotherapy effectiveness in advanced endometrial cancer, stratified by Tumor Mutational Burden (TMB) ≥10 mut/MB and microsatellite instability (MSI). METHODS: Patients with advanced endometrial cancer in the US-based de-identified Flatiron Health-Foundation Medicine Clinico-Genomic Database were included. Data originated from patients treated between January 2011- March 2022 at 280 US clinics. Next-generation sequencing assays were performed via FoundationOne or FoundationOneCDx. Longitudinal clinical data were derived from electronic health records. Immune checkpoint inhibitor treatment included pembrolizumab, dostarlimab, and nivolumab monotherapies. Time to next treatment, time to treatment discontinuation, and overall survival were assessed with the log-rank test and Cox proportional hazard models with adjusted hazard ratios (aHR) for known prognostic factors. We used the Likelihood ratio test to compare biomarker performance. RESULTS: A total of 343 patients received chemotherapy and 28 received immune checkpoint inhibitor monotherapy as frontline treatment. Patients who received monotherapy were more likely to be stage III at diagnosis (immune checkpoint inhibitor: 54.6% vs chemotherapy: 15.0%; p<0.001) and more likely to test MSI-high via next-generation sequencing (immune checkpoint inhibitor: 53.6% vs chemotherapy: 19.2%; p<0.001). In MSI-high cancers, single-agent immune checkpoint inhibitor had a more favorable time to next treatment (aHR: 0.18, p=0.001) and overall survival (aHR 0.29, p=0.045). Additional analyses on 70 unique tumor specimens revealed mismatch repair deficiency (dMMR) via immunohistochemistry and MSI-high via next-generation sequencing concordance (91%), with nominal improvement of MSI over dMMR to predict time to treatment discontinuation (p=0.030), time to next treatment (p=0.032), and overall survival (p=0.22). MSI status was concordant with tumor mutational burden ≥10 in 94.3% of cases. CONCLUSION: Immune checkpoint inhibitors may have improved efficacy over chemotherapy in frontline treatment for advanced endometrial cancer defined by MSI-high using next-generation sequencing as a nominally better predictor of outcomes than dMMR with immunohistochemistry. This provides the biologic rationale of active phase III trials.

5-FU-miR-15a Inhibits Activation of Pancreatic Stellate Cells by Reducing YAP1 and BCL-2 Levels In Vitro.

Chronic pancreatitis is characterized by chronic inflammation and fibrosis, processes heightened by activated pancreatic stellate cells (PSCs). Recent publications have demonstrated that miR-15a, which targets YAP1 and BCL-2, is significantly downregulated in patients with chronic pancreatitis compared to healthy controls. We have utilized a miRNA modification strategy to enhance the therapeutic efficacy of miR-15a by replacing uracil with 5-fluorouracil (5-FU). We demonstrated increased levels of YAP1 and BCL-2 (both targets of miR-15a) in pancreatic tissues obtained from Ptf1aCreERTM and Ptf1aCreERTM;LSL-KrasG12D mice after chronic pancreatitis induction as compared to controls. In vitro studies showed that delivery of 5-FU-miR-15a significantly decreased viability, proliferation, and migration of PSCs over six days compared to 5-FU, TGFß1, control miR, and miR-15a. In addition, treatment of PSCs with 5-FU-miR-15a in the context of TGFß1 treatment exerted a more substantial effect than TGFß1 alone or when combined with other miRs. Conditioned medium obtained from PSC cells treated with 5-FU-miR-15a significantly inhibits the invasion of pancreatic cancer cells compared to controls. Importantly, we demonstrated that treatment with 5-FU-miR-15a reduced the levels of YAP1 and BCL-2 observed in PSCs. Our results strongly suggest that ectopic delivery of miR mimetics is a promising therapeutic approach for pancreatic fibrosis and that 5-FU-miR-15a shows specific promise.

The use of andexanet alfa and 4-factor prothrombin complex concentrate in intracranial hemorrhage.

OBJECTIVE: to describe the clinical and safety outcomes between andexanet alfa (AA) and 4-factor prothrombin complex concentrate (4F-PCC) for the reversal of apixaban or rivaroxaban in the setting of an intracranial hemorrhage (ICH). METHODS: A retrospective, multicentered descriptive study was conducted in hospitalized patients 18 years of age or older from June 2018 to October 2019 who received AA or 4F-PCC for the reversal of apixaban or rivaroxaban in the setting of ICH. Patients were excluded if they had received 4F-PCC prior to AA after its addition to the institution wide formulary. Other exclusion criteria were history or presence of heparin-induced thrombocytopenia or disseminated intravascular coagulation, estimated hematoma volume of >60 mL, Glasgow Coma Scores <7, or no repeat CT head scan. Information was collected from the electronic medical records. The primary outcome was the achievement of excellent or good hemostatic efficacy upon the repeat computer tomography (CT) scan performed after the infusion of study drugs. Secondary outcomes included disposition, survival to hospital discharge, 30-day readmission, length of hospital stay, length of ICU stay, incidence of thromboembolic events. RESULTS: A total of 24 patients were included in the study, of which 9 received AA and 15 received 4F-PCC. The achievement of excellent or good hemostatic efficacy upon repeat CT scan occurred in 7 (77.8%) patients in the AA group and in 14 (93.3%) patients in the 4-F PCC group. All patients in the AA group survived to hospital discharge with no 30-day morality and 86.7% patients in the 4F-PCC group. CONCLUSION: This study suggests that real-world clinical and safety outcomes between andexanet alfa and 4F-PCC for the reversal of factor Xa inhibitors in the setting of ICH are similar to ones reported in clinical trials.

A Comparison of Topical Agents for Eschar Removal in a Porcine Model: Bromelain-enriched vs Traditional Collagenase Agents.

Surgical excision and grafting of deep partial-thickness (DPT) and full-thickness (FT) burns is a cornerstone of wound care. The use of commercially available topical enzymatic agents has been limited due to slower and less complete eschar removal than surgical excision. Using a porcine model of DPT and FT burns, we compared the eschar removal efficacy of a bromelain-enriched enzymatic agent derived from the stems of pineapple plants and a commercially available collagenase. We created 40 DPT and 40 FT burns on four anesthetized Yorkshire pigs. Eschar removal was initiated 24 hours later. Two pigs each were randomly assigned to collagenase or the bromelain-enriched agent. The bromelain-enriched agent was applied topically once for 4 hours followed by a 2-hour soaking. The collagenase was applied topically daily until complete removal of eschar or for up to 14 days. All bromelain-enriched treated FT burns underwent complete removal of the eschar after a single application while none of the collagenase-treated FT burns underwent complete removal of the eschar even after 14 days of treatment. All bromelain-enriched treated DPT burns had complete eschar removal after the single application. None of the collagenase-treated DPT burns experienced complete removal of eschar after 10 days; by day 14, 35% had complete eschar removal, 30% had >50% eschar removed, and 35% had <50% eschar removed. We conclude that eschar removal is quicker and more complete with the bromelain-enriched compared with collagenase debriding agent.

Discovery of Anthranilic Acid Derivatives as Difluoromethylornithine Adjunct Agents That Inhibit Far Upstream Element Binding Protein 1 (FUBP1) Function.

Polyamine biosynthesis is regulated by ornithine decarboxylase (ODC), which is transcriptionally activated by c-Myc. A large library was screened to find molecules that potentiate the ODC inhibitor, difluoromethylornithine (DFMO). Anthranilic acid derivatives were identified as DFMO adjunct agents. Further studies identified the far upstream binding protein 1 (FUBP1) as the target of lead compound 9. FUBP1 is a single-stranded DNA/RNA binding protein and a master controller of specific genes including c-Myc and p21. We showed that 9 does not inhibit 3H-spermidine uptake yet works synergistically with DFMO to limit cell growth in the presence of exogenous spermidine. Compound 9 was also shown to inhibit the KH4 FUBP1-FUSE interaction in a gel shift assay, bind to FUBP1 in a ChIP assay, reduce both c-Myc mRNA and protein expression, increase p21 mRNA and protein expression, and deplete intracellular polyamines. This promising hit opens the door to new FUBP1 inhibitors with increased potency.

Use of Patient-Reported Outcomes for Assessing Diabetes Outcomes.

The treatment of diabetes can be complex and overwhelming for patients as it demands persistent attention to lifestyle management, adherence to medications, monitoring of side effects of drugs, and management of devices for glucose monitoring and/or insulin infusion. Therefore, understanding patient-reported outcomes (PROs) that provide direct insight into the patient's experience with diabetes is crucial for optimizing diabetes management.This review provides an overview of commonly used PRO questionnaires that assess different aspects of diabetes management.

Awareness of the Malaria Vaccine in India.

BACKGROUND: Malaria remains a serious public health problem in India. According to the World Health Organization (WHO), as per the 2021 report, India accounts for 83% of malaria cases in Southeast Asia. Various interventions have been implemented to control malaria's burden in India. In October 2021, the WHO approved the RTS,S/AS01 (RTS,S) malaria vaccine for administration in four scheduled doses in children five months of age to reduce the burden and severity of malaria. The objectives of this study were to assess public awareness about the vaccine among residents of India and determine any associations with demographic characteristics. METHODS: The study was a web-based, cross-sectional survey. The survey questionnaire was sent out electronically using Qualtrics® (Provo, UT) and remained active for 12 weeks (December 2021 to March 2022). The questionnaire was self-administered anonymously, using a link that was shared with people across India through social media platforms. A total of 2,371 respondents above 18 years of age and current residents of India participated in the study. The chi-square test was used to examine the association between awareness about the vaccine and demographic characteristics. A p-value of <0.05 was used to describe a statistically significant association. RESULTS: Most participants (71.95%) had heard about the malaria vaccine, and 68.75% favored making it a required childhood vaccine. Similarly, 67.27% indicated that they would encourage caregivers to get their children/wards vaccinated. Age, gender, educational status, residence, and caregiver status were associated with the awareness regarding the malaria vaccine (p < 0.05). Males, participants between 18 and 24 years old, and caregivers of children aged five years or less were more likely to be aware of the vaccine. Participants with higher education and residing in urban localities had more awareness of the vaccine. CONCLUSION: The malaria vaccine has the potential to eradicate malaria in India, especially if included in the immunization schedule for children. However, it is critical that health policymakers target populations that are less aware of information on any intended rollout of the malaria vaccine to ensure rapid uptake toward the goal of eliminating malaria from India.

Ambulation Control System Design for a Hybrid Knee Prosthesis.

Prosthetic knees available to individuals with transfemoral amputation seek to restore functional ability to the user. Passive prosthetic knees are lightweight but can restore only limited, dissipative ambulation activities whereas active knees can provide energy to restore additional ambulation activities such as stair climbing and standing up from a chair. Semi-active prosthetic devices aim to only power a subset of activities and use passive components and control when that power is not necessary. Here, we outline an ambulation control system for a lightweight Hybrid Knee prosthesis that is powered for climbing stairs and passive for other ambulation activities (level-ground walking, walking on an incline, and stair descent). We include preliminary offline and online intent recognition system results for one able-bodied user and one individual with a transfemoral amputation demonstrating low error rates in predicting between active and passive control.

An Oncological Emergency: Severe Type B Lactic Acidosis From Warburg Effect in Diffuse Large B-cell Lymphoma.

Lactic acidosis is the most common anion gap metabolic acidosis in critically ill patients. Type B lactic acidosis is most commonly seen with hematological malignancies, especially lymphomas. It is considered an oncological emergency and is associated with high mortality and poor outcomes if not treated promptly. Here, we present the case of a 48-year-old male who developed Type B lactic acidosis secondary to newly diagnosed diffuse large B-cell lymphoma. This case highlights the importance of including Type B lactic acidosis in the differential diagnosis in a patient with unexplained lactic acidosis and hypoglycemia with otherwise vague symptoms and the need for a thorough search for quick diagnosis and early management.

Diversity of two-pore channels and the accessory NAADP receptors in intracellular Ca2+ signaling.

Intracellular Ca2+ signaling via changes or oscillation in cytosolic Ca2+ concentration controls almost every aspect of cellular function and physiological processes, such as gene transcription, cell motility and proliferation, muscle contraction, and learning and memory. Two-pore channels (TPCs) are a class of eukaryotic cation channels involved in intracellular Ca2+ signaling, likely present in a multitude of organisms from unicellular organisms to mammals. Accumulated evidence indicates that TPCs play a critical role in Ca2+ mobilization from intracellular stores mediated by the second messenger molecule, nicotinic acid adenine dinucleotide phosphate (NAADP). In recent years, significant progress has been made regarding our understanding of the structures and function of TPCs, including Cryo-EM structure determination of mammalian TPCs and characterization of a plastid TPC in a single-celled parasite.. The recent identification of Lsm12 and JPT2 as NAADP-binding proteins provides a new molecular basis for understanding NAADP-evoked Ca2+ signaling. In this review, we summarize basic structural and functional aspects of TPCs and highlight the most recent studies on the newly discovered TPC in a parasitic protozoan and the NAADP-binding proteins LSM12 and JPT2 as new key players in NAADP signaling.

Increased foot strike variability during turning in Parkinson's disease patients with freezing of gait.

BACKGROUND: Turning is a common trigger for freezing episodes in patients with Parkinson's disease (PD). Freezing during turning can lead to falls and fractures and decreased quality of life. RESEARCH QUESTION: Does foot-strike contact variability also increase during turning, as previously reported in straight gait in PD patients with Freezing of Gait (FOG)? METHODS: Subjects were instructed to walk on a gait mat, making "normal pivot" (180°) turns at each end. ProtoKinetics Movement Analysis Software (PKMAS) software was used for analysis. Video recordings and foot-pressure-prints were studied to identify and define turn segments. Spatiotemporal gait and turn measures were then determined only for the turn segments. A movement disorders neurologist determined clinical freezes. RESULTS: 100 subjects (28 controls, 38 noFOG and 34 FOG) were included. Compared to non-freezers (noFOG), FOG subjects had a smaller foot-strike during turning (a measure of completeness of foot contact with the mat) and increased foot-strike variability. FOG subjects also had a shorter stride-length, slower stride-velocity, and greater swing phase time and percentage during turns. After adjusting for turn direction, inner/outer leg dynamics showed heavier inner leg footsteps in FOG subjects. 38% of FOG subjects experienced freezes during turning. 69% of freezes occurred during the middle third of the turn. Turn-freezers had more severe spatiotemporal gait deficits. SIGNIFICANCE: Developing targeted therapies to retrain subjects to plant their whole foot on the ground with more consistency could help decrease episodes of freezing of gait.

Targeted therapy for LIMD1-deficient non-small cell lung cancer subtypes.

An early event in lung oncogenesis is loss of the tumour suppressor gene LIMD1 (LIM domains containing 1); this encodes a scaffold protein, which suppresses tumorigenesis via a number of different mechanisms. Approximately 45% of non-small cell lung cancers (NSCLC) are deficient in LIMD1, yet this subtype of NSCLC has been overlooked in preclinical and clinical investigations. Defining therapeutic targets in these LIMD1 loss-of-function patients is difficult due to a lack of 'druggable' targets, thus alternative approaches are required. To this end, we performed the first drug repurposing screen to identify compounds that confer synthetic lethality with LIMD1 loss in NSCLC cells. PF-477736 was shown to selectively target LIMD1-deficient cells in vitro through inhibition of multiple kinases, inducing cell death via apoptosis. Furthermore, PF-477736 was effective in treating LIMD1-/- tumours in subcutaneous xenograft models, with no significant effect in LIMD1+/+ cells. We have identified a novel drug tool with significant preclinical characterisation that serves as an excellent candidate to explore and define LIMD1-deficient cancers as a new therapeutic subgroup of critical unmet need.