A blood-brain barrier-penetrant AAV gene therapy improves neurological function in symptomatic mucolipidosis IV mice.

Mucolipidosis IV (MLIV) is a rare, autosomal recessive, lysosomal disease characterized by intellectual disability, motor deficits, and progressive vision loss. Using adeno-associated vector 9 (AAV9) and AAV-PHP.B as delivery vectors, we previously demonstrated the feasibility of modifying disease course in a mouse model of MLIV by the human MCOLN1 gene transfer. Here, using a primate-enabling capsid AAV.CPP.16 (CPP16), we constructed a new, clinic-oriented MCOLN1 gene expression vector and demonstrated its efficacy in the preclinical model of MLIV. Systemic administration of CPP16-MCOLN1 in adult symptomatic Mcoln1 -/- mice at a dose of 1e12 vg per mouse resulted in MCOLN1 expression in the brain and peripheral tissues, alleviated brain pathology, rescued neuromotor function, and completely prevented paralysis. Notable expression of MCOLN1 transcripts was also detected in the retina of the mouse, which had exhibited significant degeneration at the time of the treatment. However, no increase in retinal thickness was observed after gene therapy treatment. Our results suggest a new AAV-based systemic gene replacement therapy for the treatment of MLIV that could be translated into clinical studies.

Interrater Agreement of BT-RADS for Evaluation of Follow-Up MRI in Treated Primary Brain Tumor Patients.

BACKGROUND AND PURPOSE: The Brain Tumor Reporting and Data System (BT-RADS) is a structured radiology reporting algorithm that was introduced to provide uniformity in post-treatment primary brain tumor follow-up and reporting, but its interrater reliability (IRR) assessment has not been widely studied. Our goal is to evaluate the IRR among neuroradiologists and radiology residents in the use of BT-RADS. MATERIALS AND METHODS: This retrospective study reviewed 103 consecutive MR studies in 98 adult patients previously diagnosed with and treated for primary brain tumor (January 2019 to February 2019). Six readers with varied experience (4 neuroradiologists and 2 radiology residents) independently evaluated each case and assigned a BT-RADS score. Readers were blinded to the original score reports and the reports from other readers. Cases in which at least one neuroradiologist scored differently were subjected to consensus scoring. After the study, a post-hoc reference score was also assigned by 2 readers using future imaging and clinical information previously unavailable to readers. The interrater reliabilities were assessed using Gwet's AC2 index with ordinal weights and percent agreement. RESULTS: Of the 98 patients evaluated (median age, 53 years; interquartile range, 41-66 years), 53% were males. The most common tumor type was astrocytoma (77%) of which 56% were grade 4 glioblastoma. Gwet's index for interrater reliability among all six readers was 0.83 (95% CI: 0.78, 0.87). The Gwet's index for the neuroradiologists' group (0.84 [95% CI: 0.79, 0.89]) was not statistically different from that for the residents' group (0.79 [95% CI: 0.72, 0.86]) (χ2 = 0.85; p = 0.36). All four neuroradiologists agreed on the same BT-RADS score in 57 of the 103 studies, three neuroradiologists agreed in 21 of the 103 studies, and two neuroradiologists agreed in 21 of the 103 studies. Percent agreement between neuroradiologist blinded scores and post-hoc reference scores ranged from 41%-52%. CONCLUSIONS: A very good interrater agreement was found when tumor reports were interpreted by independent blinded readers using BT-RADS criteria. Further study is needed to determine if this high overall agreement can translate into greater consistency in clinical care. ABBREVIATIONS: BI-RADS = Breast Imaging Reporting and Data System; BT-RADS = Brain Tumor Reporting and Data System; IQR = interquartile range; IRR = interrater reliability; NI-RADS = Neck Imaging Reporting and Data System.

Craniodental ecomorphology of the large Jurassic ichthyosaurian Temnodontosaurus.

Marine amniotes have played many crucial roles in ocean ecosystems since the Triassic, including predation at the highest trophic levels. One genus often placed into this guild is the large Early Jurassic neoichthyosaurian Temnodontosaurus, the only post-Triassic ichthyosaurian known with teeth which bear a distinct cutting edge or carina. This taxonomically problematic genus is currently composed of seven species which show a wide variety of skull and tooth morphologies. Here we assess the craniodental disparity in Temnodontosaurus using a series of functionally informative traits. We describe the range of tooth morphologies in the genus in detail, including the first examples of serrated carinae in ichthyosaurians. These consist of false denticles created by the interaction of enamel ridgelets with the carinal keel, as well as possible cryptic true denticles only visible using scanning electron microscopy. We also find evidence for heterodonty in the species T. platyodon, with unicarinate mesial teeth likely playing a role in prey capture and labiolingually compressed, bicarinate distal teeth likely involved in prey processing. This type of heterodonty appears to be convergent with a series of other marine amniotes including early cetaceans. Overall, the species currently referred to as the genus Temnodontosaurus show a range of craniodental configurations allowing prey to be captured and processed in different ways - for example, T. eurycephalus has a deep snout and relatively small bicarinate teeth likely specialised for increased wound infliction and grip-and-tear feeding, whereas T. platyodon has a more elongate yet robust snout and larger teeth and may be more adapted for grip-and-shear feeding. These results suggest the existence of niche partitioning at higher trophic levels in Early Jurassic ichthyosaurians and have implications for future work on the taxonomy of this wastebasket genus, as well as for research into the ecology of other extinct megapredatory marine tetrapods.

Total Knee Arthroplasty in the Valgus Knee.

An estimated 10 to 15% of total knee arthroplasties (TKAs) are implanted for a diagnosis of arthritis when a valgus deformity is present. There are various techniques and considerations that must be considered for a successful TKA in a patient with a valgus deformity. This article provides a detailed summary of the anatomy, pathology, bone preparation, soft tissue management, implant selection, and complications when performing a TKA in a patient with valgus deformity.

Building an Extracorporeal Cardiopulmonary Resuscitation Program at a High-volume Extracorporeal Membrane Oxygenation Center.

Extracorporeal Cardiopulmonary Resuscitation (ECPR) is an emerging approach to cardiac arrest. We present two contrasting cases from a high-volume extracorporeal membrane oxygenation (ECMO) center (defined as greater than 30 ECMO cases per year) without a 24/7 ECPR program to highlight how to establish an ECPR program with a focus on patient selection and outcome optimization. In one case, a patient presented with cardiac arrest during initial triage for chest pain within the emergency department, and in the other case, a patient experienced an out-of-hospital cardiac arrest with prolonged no-flow and low-flow time. Despite the lack of a 24/7 ECPR program at the presenting center, both patients received an ECPR evaluation, as both patients presented while all services necessary for ECMO cannulation were available. The in-hospital cardiac arrest patient was successfully cannulated for ECMO during cardiopulmonary resuscitation and survived with few complications. The out-of-hospital cardiac arrest patient was deemed a poor candidate for ECPR and expired soon after presentation. These two cases highlight the complex decision-making in ECPR and further illustrate how to create ECPR protocols at a high-volume ECMO center before resources are available for a 24/7 ECPR program.

Primary Care Physician Supply and Population Health Outcomes in Florida, 2010-2019.

Background: Primary care physicians play vital roles in the prevention and management of chronic disease. With increasing rates of chronic disease and a national primary care physician shortage, the role that primary care physician supply has on health outcomes in Florida is not well understood. The objective of this study was to investigate the relationship between primary care physician supply (PCPS) and population health outcomes of obesity, life expectancy, coronary artery disease hospitalization, and death rate as reported by county in the state of Florida for the years 2010, 2013, 2016, and 2019. Methods: This was a retrospective, cross-sectional study. Secondary data was used from the Florida Department of Health. Numerous population health and social determinants of health variables related to PCPS in the literature were selected for analysis. Correlation and linear regression analyses were conducted using STATA14. Results: The association between PCPS and obesity was the strongest association in this analysis and was significant for each year with an average of 9.25 primary care physicians per 100 000 people needed to decrease the obesity rate by 1%. PCPS was positively correlated with life expectancy for years 2013, 2016, and 2019 and negatively correlated with the death rate in 2010 and 2019. In the multiple regression, PCPS was negatively associated with areas having a high rate of uninsured persons, unemployment, decreased education, and age over 65. Conclusion: Increased supply of primary care physicians in Florida is significantly associated with decreased rates of obesity and death and increased life expectancy. Our results also indicate that areas with higher levels of social vulnerability also have inequitable distributions of PCPS. Therefore, PCPS should be increased, particularly in areas with the highest need, as primary care physicians in the state of Florida play an important role in improving the overall health of the populations they serve.

Data-Driven Approach for Upper Limb Fatigue Estimation Based on Wearable Sensors.

Muscle fatigue is defined as a reduced ability to maintain maximal strength during voluntary contraction. It is associated with musculoskeletal disorders that affect workers performing repetitive activities, affecting their performance and well-being. Although electromyography remains the gold standard for measuring muscle fatigue, its limitations in long-term work motivate the use of wearable devices. This article proposes a computational model for estimating muscle fatigue using wearable and non-invasive devices, such as Optical Fiber Sensors (OFSs) and Inertial Measurement Units (IMUs) along the subjective Borg scale. Electromyography (EMG) sensors are used to observe their importance in estimating muscle fatigue and comparing performance in different sensor combinations. This study involves 30 subjects performing a repetitive lifting activity with their dominant arm until reaching muscle fatigue. Muscle activity, elbow angles, and angular and linear velocities, among others, are measured to extract multiple features. Different machine learning algorithms obtain a model that estimates three fatigue states (low, moderate and high). Results showed that between the machine learning classifiers, the LightGBM presented an accuracy of 96.2% in the classification task using all of the sensors with 33 features and 95.4% using only OFS and IMU sensors with 13 features. This demonstrates that elbow angles, wrist velocities, acceleration variations, and compensatory neck movements are essential for estimating muscle fatigue. In conclusion, the resulting model can be used to estimate fatigue during heavy lifting in work environments, having the potential to monitor and prevent muscle fatigue during long working shifts.

Management of open wounds in dogs using a bioresorbable polymeric wound matrix: 14 Cases (2019-2021).

The objective of this retrospective case series is to evaluate clinical outcomes associated with the use of a bioresorbable polymeric wound matrix (MicroLyte Vet; Imbed Biosciences) for the management of soft tissue wounds of various etiologies in dogs. Medical records were reviewed between August 2019 and November 2021 for signalment, wound etiology, location, size, initial wound management, antibiotic therapy, time to initial application, number of applications, time between applications, secondary dressing used, time to wound closure, complications, and clinical outcomes. Time to initial application ranged from 1 to 61 days (median, 7.5). The number of applications ranged from 1 to 15 (median, 4.5). Time between applications ranged from 1 to 8 days (median, 3.7). Additional surgical interventions were implemented in 3 dogs. Wound closure by second intention healing (n=14) ranged from 2 to 12 weeks (median, 5). There were no adverse events attributed to the use of bioresorbable polymeric wound matrix. Complete wound healing occurred with use of the bioresorbable polymeric wound matrix in all dogs.

Diagnostic Excellence in Pediatric Spine Imaging: Using Contextualized Imaging Protocols.

Contextual design and selection of MRI protocols is critical for making an accurate diagnosis given the wide variety of clinical indications for spine imaging in children. Here, we describe our pediatric spine imaging protocols in detail, tailored to specific clinical questions.

A fabric-based soft hand exoskeleton for assistance: the ExHand Exoskeleton.

Introduction: The rise of soft robotics has driven the development of devices for assistance in activities of daily living (ADL). Likewise, different types of actuation have been developed for safer human interaction. Recently, textile-based pneumatic actuation has been introduced in hand exoskeletons for features such as biocompatibility, flexibility, and durability. These devices have demonstrated their potential use in assisting ADLs, such as the degrees of freedom assisted, the force exerted, or the inclusion of sensors. However, performing ADLs requires the use of different objects, so exoskeletons must provide the ability to grasp and maintain stable contact with a variety of objects to lead to the successful development of ADLs. Although textile-based exoskeletons have demonstrated significant advancements, the ability of these devices to maintain stable contact with a variety of objects commonly used in ADLs has yet to be fully evaluated. Materials and methods: This paper presents the development and experimental validation in healthy users of a fabric-based soft hand exoskeleton through a grasping performance test using The Anthropomorphic Hand Assessment Protocol (AHAP), which assesses eight types of grasping with 24 objects of different shapes, sizes, textures, weights, and rigidities, and two standardized tests used in the rehabilitation processes of post- stroke patients. Results and discussion: A total of 10 healthy users (45.50 ± 14.93 years old) participated in this study. The results indicate that the device can assist in developing ADLs by evaluating the eight types of grasps of the AHAP. A score of 95.76 ± 2.90% out of 100% was obtained for the Maintaining Score, indicating that the ExHand Exoskeleton can maintain stable contact with various daily living objects. In addition, the results of the user satisfaction questionnaire indicated a positive mean score of 4.27 ± 0.34 on a Likert scale ranging from 1 to 5.

Soft-Sensor System for Grasp Type Recognition in Underactuated Hand Prostheses.

This paper presents the development of an intelligent soft-sensor system to add haptic perception to the underactuated hand prosthesis PrHand. Two sensors based on optical fiber were constructed, one for finger joint angles and the other for fingertips' contact force. Three sensor fabrications were tested for the angle sensor by axially rotating the sensors in four positions. The configuration with the most similar response in the four rotations was chosen. The chosen sensors presented a polynomial response with R2 higher than 92%. The tactile force sensors tracked the force made over the objects. Almost all sensors presented a polynomial response with R2 higher than 94%. The system monitored the prosthesis activity by recognizing grasp types. Six machine learning algorithms were tested: linear regression, k-nearest neighbor, support vector machine, decision tree, k-means clustering, and hierarchical clustering. To validate the algorithms, a k-fold test was used with a k = 10, and the accuracy result for k-nearest neighbor was 98.5%, while that for decision tree was 93.3%, enabling the classification of the eight grip types.

All-in-One digital microfluidics pipeline for proteomic sample preparation and analysis.

Highly sensitive and reproducible analysis of samples containing low amounts of protein is restricted by sample loss and the introduction of contaminants during processing. Here, we report an All-in-One digital microfluidic (DMF) pipeline for proteomic sample reduction, alkylation, digestion, isotopic labeling and analysis. The system features end-to-end automation, with integrated thermal control for digestion, optimized droplet additives for sample manipulation and analysis, and an automated interface to liquid chromatography with tandem mass spectrometry (HPLC-MS/MS). Dimethyl labeling was integrated into the pipeline to allow for relative quantification of the trace samples at the nanogram level, and the new pipeline was applied to evaluating cancer cell lines and cancer tissue samples. Several known proteins (including HSP90AB1, HSPB1, LDHA, ENO1, PGK1, KRT18, and AKR1C2) and pathways were observed between model breast cancer cell lines related to hormone response, cell metabolism, and cell morphology. Furthermore, differentially quantified proteins (such as PGS2, UGDH, ASPN, LUM, COEA1, and PRELP) were found in comparisons of healthy and cancer breast tissues, suggesting potential utility of the All-in-One pipeline for the emerging application of proteomic cancer sub-typing. In sum, the All-in-One pipeline represents a powerful new tool for automated proteome processing and analysis, with the potential to be useful for evaluating mass-limited samples for a wide range of applications.

Classifying Bone Loss in Failed Stemmed Total Knee Arthroplasty: Determining Reliability.

BACKGROUND: Failed stemmed total knee arthroplasty (TKA) components present with varying degrees of bone loss and technical challenges. A classification system has been proposed based upon metaphyseal bone loss and diaphyseal cortical integrity. A validation study was performed to determine interobserver and intraobserver reliability at multiple institutions and with different levels of training. METHODS: An online survey with digital anteroposterior and lateral radiographs was sent to 5 arthroplasty surgeons and 5 adult reconstruction fellows. The survey included 62 cases with stemmed femoral and tibial components, considered failures and pending revision, and scored by each reviewer independently using the classification system. Each case was scored in 2 separate sessions. Interobserver and intraobserver reliability was assessed using the intraclass correlation coefficient (ICC). RESULTS: Interobserver grading for both the femur (0.69) and tibia (0.72) showed strong reliability among the attendings and fellows, with slightly stronger reliability in tibia cases. The intraclass correlation coefficient (ICC) for attendings and fellows was similar overall, demonstrating consistency of the grading regardless of training level. Intraobserver comparisons showed a strong ICC for attendings and fellows in femoral cases, while fellows had near-perfect ICC in tibia cases. Across all reviewers there was on average 93% agreement within 1 grade per case with the majority of the discrepancy occurring at the metaphyseal-diaphyseal junction. CONCLUSION: This classification demonstrated overall strong interobserver and intraobserver reliability, with 93% agreement within 1 grade of bone loss. With further education, this classification system can ultimately be used to standardize the degree of bone loss in failed stemmed components and help with preoperative planning.

Femoral stem migration after direct lateral and direct anterior total hip arthroplasty: a prospective cohort study.

BACKGROUND: The direct lateral (DL) approach to total hip arthroplasty is more commonly used than the newer direct anterior (DA) approach. Both approaches use collared or collarless femoral stems. We sought to assess implant stability of stem designs implanted with the DL approach and compare outcomes from this cohort with those of a previously reported cohort of patients who underwent arthroplasty with a DA approach. We also sought to determine if early recovery influences differences in migration. METHODS: Patients underwent total hip arthroplasty using the DL or the DA approach and were randomized to receive either a collared or collarless, cementless femoral stem. On the day of surgery and at 6 follow-up visits through to 1 year, patients underwent supine radiostereometric imaging to track implant migration. At follow-up visits, patients performed an instrumented walking test to assess their functional ability and logged an average daily step count to assess their activity levels. We assessed whether patient function and activity were correlated with migration. RESULTS: Stem design did not have a significant effect on migration for the DL group (p = 0.894). Compared with the DA group, the DL group migrated significantly less for both collared (p = 0.031) and collarless (p = 0.002) stems. Migration was not correlated with function or activity at any time point (p > 0.05). CONCLUSION: Most implant migration occurred from the day of surgery to 2 weeks after the operation and stabilized thereafter, suggesting adequate fixation and a low risk for aseptic loosening in both patient groups.

Chronic myocardial and coronary arterial effects of intracoronary supersaturated oxygen therapy in swine with normal and ischemic-reperfused myocardium.

The study assessed chronic myocardial, coronary and systemic effects of intracoronary supersaturated oxygen (SSO2) therapy. Left anterior descending coronary arteries of 40 swine were stented and randomized to 90-min selective intracoronary infusion of SSO2 (pO2 760-1000 mmHg) or normoxemic saline. In 20 out of 40 animals, SSO2 delivery followed a 60-min balloon occlusion to induce myocardial infarction (MI). In both normal and MI models, intracoronary treatment with hyperoxemic SSO2 therapy showed no evidence of coronary thrombosis. There were no biologically relevant differences between treatments at either time point in regard to coronary intervention site healing and neointimal growth. No signs of any myocardial or systemic toxicity were observed after 7 or 30 days. A trend was observed toward reduced incidence of microscopic MI scars and reduced infarct size in histopathology, as well as toward better recovery of echocardiographically evaluated global and regional contractility at 30 days. No treatment related infarcts or thromboemboli were observed in the downstream organs.

Definition and time course of pericavity edema after minimally invasive endoscopic intracerebral hemorrhage evacuation.

BACKGROUND: Perihematomal edema (PHE) volume correlates with intracerebral hemorrhage (ICH) volume and is associated with functional outcome. Minimally invasive surgery (MIS) for ICH decreases clot burden and PHE. MIS may therefore alter the time course of PHE, mitigating a critical source of secondary injury. OBJECTIVE: To describe a new method for the quantitative measurement of cerebral edema surrounding the evacuated hematoma cavity, termed pericavity edema (PCE), and obtain details of its time course following MIS for ICH. METHODS: The study included 48 consecutive patients presenting with ICH who underwent MIS evacuation. Preoperative and postoperative CT scans were assessed by two independent raters. Hematoma, edema, cavity, and pneumocephalus volumes were calculated using semi-automatic, threshold-guided volume segmentation software (AnalyzePro). Follow-up CT scans at variable delayed time points were available for 36 patients and were used to describe the time course of PCE. RESULTS: Mean preoperative, postoperative, and delayed PCE were 21.0 mL (SD 15.5), 18.6 mL (SD 11.4), and 18.4 mL (SD 15.5), respectively. The percentage of ICH evacuated correlated significantly with a decrease in postoperative PCE (r=-0.46, p<0.01). Linear regression analysis revealed a significant relation between preoperative hematoma volume and both postoperative PCE (p<0.001) and postoperative relative PCE (p<0.001). The mean peak PCE was 26.4 mL (SD 15.6) and occurred at 6.5 days (SD 4.8) post-ictus. The 2-week postoperative time course of relative PCE did not fluctuate, suggesting stability in edema during the perioperative period surrounding evacuation and up to 2 weeks after the initial bleed. CONCLUSIONS: We present a detailed and accurate method for measuring PCE volume with semi-automatic, threshold-guided segmentation software in the postoperative patient with ICH. Decrease in PCE after MIS evacuation correlated with evacuation percentage, and relative PCE remained stable after minimally invasive endoscopic ICH evacuation.

Gene family evolution underlies cell-type diversification in the hypothalamus of teleosts.

Hundreds of cell types form the vertebrate brain but it is largely unknown how similar cellular repertoires are between or within species or how cell-type diversity evolves. To examine cell-type diversity across and within species, we performed single-cell RNA sequencing of ~130,000 hypothalamic cells from zebrafish (Danio rerio) and surface and cave morphs of Mexican tetra (Astyanax mexicanus). We found that over 75% of cell types were shared between zebrafish and Mexican tetra, which diverged from a common ancestor over 150 million years ago. Shared cell types displayed shifts in paralogue expression that were generated by subfunctionalization after genome duplication. Expression of terminal effector genes, such as neuropeptides, was more conserved than the expression of their associated transcriptional regulators. Species-specific cell types were enriched for the expression of species-specific genes and characterized by the neofunctionalization of expression patterns of members of recently expanded or contracted gene families. Comparisons between surface and cave morphs revealed differences in immune repertoires and transcriptional changes in neuropeptidergic cell types associated with genomic differences. The single-cell atlases presented here are a powerful resource to explore hypothalamic cell types and reveal how gene family evolution and shifts in paralogue expression contribute to cellular diversity.

An analysis of Medtronic MiniMed 670G insulin pump use in clinical practice and the impact on glycemic control, quality of life, and compliance.

AIMS: This study evaluated the use of the Medtronic MiniMed 670G system in adults with type 1 diabetes mellitus from a large endocrinology practice and its impact on glycemic control, quality of life (QoL), compliance and safety. METHODS: 84 participants completed one site visit for data collection. Percentage of time in range (TIR: 70-180 mg/dL), hyperglycemia or time above range (TAB) (>180 mg/dL), hypoglycemia or time below range (TBR) (<70 mg/dL), HbA1c, average blood glucose (ABG), and other metrics were evaluated at the last visit using the system (LVMM) and compared between the last visit on previous insulin therapy (LVPT). RESULTS: The mean percentage of TIR at the LVMM was 74.0 ± 12.1%, with an increase of 27.1% (p < 0.001) in TIR from the LVPT. The mean percentage of TAR was 22.9 ± 11.8% and the mean percentage of TBR was 3.2 ± 5.1%. CONCLUSIONS: The use of the Medtronic MiniMed 670G system in our practice resulted in a TIR above the recommended target with a high degree of treatment satisfaction and compliance in adults with type 1 diabetes. Furthermore, the system may be a reasonable choice for patients struggling with significant amounts of hypoglycemia.

Statistical learning of blunt cerebrovascular injury risk factors using the elastic net.

PURPOSE: To compare logistic regression to elastic net for identifying and ranking clinical risk factors for blunt cerebrovascular injury (BCVI). MATERIALS AND METHODS: Consecutive trauma patients undergoing screening CTA at a level 1 trauma center over a 2-year period. Each internal carotid artery (ICA) and vertebral artery (VA) was independently graded by 2 neuroradiologists using the Denver grading scale. Unadjusted odds ratios were calculated by univariate and adjusted odds ratios by multiple logistic regression with FDR correction. We applied logistic regression with the elastic net penalty and tenfold cross-validation. RESULTS: Total of 467 patients; 73 patients with BCVI. Maxillofacial fracture, basilar skull fracture, and GCS had significant unadjusted odds ratios (OR) for ICA injury and C-spine fracture, spinal ligamentous injury, and age for VA injury. Only transverse foramen fracture had significant adjusted OR for VA injury, with none for ICA injury, after FDR correction. Using elastic net, ICA injury variables included maxillofacial fracture, basilar skull fracture, GCS, and carotid canal fracture. For VA injury, these included cervical spine transverse foramen fracture, ligamentous injury, C1-C3 fractures, posterior element fracture, and vertebral body fracture. CONCLUSION: Elastic net statistical learning methods identified additional risk factors and outperformed multiple logistic regression for BCVI. Elastic net allows the study of a large number of variables, and is useful when covariates are correlated.