Lamina-Dependent Stretching and Unconventional Chromosome Compartments in Early C. elegans Embryos.

Current models suggest that chromosome domains segregate into either an active (A) or inactive (B) compartment. B-compartment chromatin is physically separated from the A compartment and compacted by the nuclear lamina. To examine these models in the developmental context of C. elegans embryogenesis, we undertook chromosome tracing to map the trajectories of entire autosomes. Early embryonic chromosomes organized into an unconventional barbell-like configuration, with two densely folded B compartments separated by a central A compartment. Upon gastrulation, this conformation matured into conventional A/B compartments. We used unsupervised clustering to uncover subpopulations with differing folding properties and variable positioning of compartment boundaries. These conformations relied on tethering to the lamina to stretch the chromosome; detachment from the lamina compacted, and allowed intermingling between, A/B compartments. These findings reveal the diverse conformations of early embryonic chromosomes and uncover a previously unappreciated role for the lamina in systemic chromosome stretching.

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.

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.

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.

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.

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.

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.

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.

Bmp signaling maintains a mesoderm progenitor cell state in the mouse tailbud.

Caudal somites are generated from a pool of progenitor cells located in the tailbud region. These progenitor cells form the presomitic mesoderm that gradually differentiates into somites under the action of the segmentation clock. The signals responsible for tailbud mesoderm progenitor pool maintenance during axial elongation are still elusive. Here, we show that Bmp signaling is sufficient to activate the entire mesoderm progenitor gene signature in primary cultures of caudal mesoderm cells. Bmp signaling acts through the key regulatory genes brachyury (T) and Nkx1-2 and contributes to the activation of several other regulators of the mesoderm progenitor gene network. In the absence of Bmp signaling, tailbud mesoderm progenitor cells acquire aberrant gene expression signatures of the heart, blood, muscle and skeletal embryonic lineages. Treatment of embryos with the Bmp inhibitor noggin confirmed the requirement for Bmp signaling for normal T expression and the prevention of abnormal lineage marker activation. Together, these results identify Bmp signaling as a non-cell-autonomous signal necessary for mesoderm progenitor cell homeostasis.

Zinc exocytosis is sensitive to myosin light chain kinase inhibition in mouse and human eggs.

Zinc dynamics are essential for oocyte meiotic maturation, egg activation, and preimplantation embryo development. During fertilisation and egg activation, the egg releases billions of zinc atoms (Zn2+) in an exocytotic event termed the 'zinc spark'. We hypothesised that this zinc transport and exocytosis is dependent upon the intracellular trafficking of cortical granules (CG) which requires myosin-actin-dependent motors. Treatment of mature mouse and human eggs with ML-7, a myosin light chain kinase inhibitor (MLCK), resulted in an 80% reduction in zinc spark intensity compared to untreated controls when activated with ionomycin. Moreover, CG migration towards the plasma membrane was significantly decreased in ML-7-treated eggs compared with controls when activated parthenogenetically with ionomycin. In sperm-induced fertilisation via intracytoplasmic sperm injection (ICSI), ML-7-treated mouse eggs exhibited decreased labile zinc intensity and cortical CG staining. Collectively, these data demonstrate that ML-7 treatment impairs zinc release from both murine and human eggs after activation, demonstrating that zinc exocytosis requires myosin light chain kinase activity. Further, these results provide additional support that zinc is likely stored and released from CGs. These data underscore the importance of intracellular zinc trafficking as a crucial component of egg maturation necessary for egg activation and early embryo development.

Modulation of apoptotic response by LAR family phosphatases-cIAP1 signaling during urinary tract morphogenesis.

The elimination of unwanted cells by apoptosis is necessary for tissue morphogenesis. However, the cellular control of morphogenetic apoptosis is poorly understood, notably the modulation of cell sensitivity to apoptotic stimuli. Ureter maturation, the process by which the ureter is displaced to the bladder wall, represents an exquisite example of morphogenetic apoptosis, requiring the receptor protein tyrosine phosphatases (RPTPs): LAR and RPTPσ. Here we show that LAR-RPTPs act through cellular inhibitor of apoptosis protein 1 (cIAP1) to modulate caspase 3,7-mediated ureter maturation. Pharmacologic or genetic inactivation of cIAP1 reverts the apoptotic deficit of LAR-RPTP-deficient embryos. Moreover, Birc2 (cIAP1) inactivation generates excessive apoptosis leading to vesicoureteral reflux in newborns, which underscores the importance of apoptotic modulation during urinary tract morphogenesis. We finally demonstrate that LAR-RPTP deficiency increases cIAP1 stability during apoptotic cell death. Together these results identify a mode of cIAP1 regulation playing a critical role in the cellular response to apoptotic pathway activation in the embryo.

Barriers to a Timely Discharge From Short-Term Care in VA Community Living Centers.

This study aimed to identify the barriers to a timely discharge from short-term care in Veterans Health Administration (VHA) Community Living Centers (CLCs). Ninety-nine interviews were conducted with CLC staff in leadership and direct-care positions in eight varied CLCs. Major themes identified through qualitative analysis as barriers to a timely discharge were a lack of patients' financial resources, low social support, and reluctance of some veterans and staff to view a timely veteran discharge as their goal. Staff also perceived that barriers were much more difficult to overcome in regions where community-based long-term services and supports were limited or nonexistent. Because VHA has lagged behind Medicaid more generally in terms of investment in these types of services, additional strategies are warranted to achieve the important policy goal of deinstitutionalizing VHA care and returning veterans to their homes in the community.

A Point Mutation in p190A RhoGAP Affects Ciliogenesis and Leads to Glomerulocystic Kidney Defects.

Rho family GTPases act as molecular switches regulating actin cytoskeleton dynamics. Attenuation of their signaling capacity is provided by GTPase-activating proteins (GAPs), including p190A, that promote the intrinsic GTPase activity of Rho proteins. In the current study we have performed a small-scale ENU mutagenesis screen and identified a novel loss of function allele of the p190A gene Arhgap35, which introduces a Leu1396 to Gln substitution in the GAP domain. This results in decreased GAP activity for the prototypical Rho-family members, RhoA and Rac1, likely due to disrupted ordering of the Rho binding surface. Consequently, Arhgap35-deficient animals exhibit hypoplastic and glomerulocystic kidneys. Investigation into the cystic phenotype shows that p190A is required for appropriate primary cilium formation in renal nephrons. P190A specifically localizes to the base of the cilia to permit axoneme elongation, which requires a functional GAP domain. Pharmacological manipulations further reveal that inhibition of either Rho kinase (ROCK) or F-actin polymerization is able to rescue the ciliogenesis defects observed upon loss of p190A activity. We propose a model in which p190A acts as a modulator of Rho GTPases in a localized area around the cilia to permit the dynamic actin rearrangement required for cilia elongation. Together, our results establish an unexpected link between Rho GTPase regulation, ciliogenesis and glomerulocystic kidney disease.

Structural basis for substrate placement by an archaeal box C/D ribonucleoprotein particle.

Box C/D small nucleolar and Cajal body ribonucleoprotein particles (sno/scaRNPs) direct site-specific 2'-O-methylation of ribosomal and spliceosomal RNAs and are critical for gene expression. Here we report crystal structures of an archaeal box C/D RNP containing three core proteins (fibrillarin, Nop56/58, and L7Ae) and a half-mer box C/D guide RNA paired with a substrate RNA. The structure reveals a guide-substrate RNA duplex orientation imposed by a composite protein surface and the conserved GAEK motif of Nop56/58. Molecular modeling supports a dual C/D RNP structure that closely mimics that recently visualized by electron microscopy. The substrate-bound dual RNP model predicts an asymmetric protein distribution between the RNP that binds and methylates the substrate RNA. The predicted asymmetric nature of the holoenzyme is consistent with previous biochemical data on RNP assembly and provides a simple solution for accommodating base-pairing between the C/D guide RNA and large ribosomal and spliceosomal substrate RNAs.

Universal mobile electrochemical detector designed for use in resource-limited applications.

This paper describes an inexpensive, handheld device that couples the most common forms of electrochemical analysis directly to "the cloud" using any mobile phone, for use in resource-limited settings. The device is designed to operate with a wide range of electrode formats, performs on-board mixing of samples by vibration, and transmits data over voice using audio--an approach that guarantees broad compatibility with any available mobile phone (from low-end phones to smartphones) or cellular network (second, third, and fourth generation). The electrochemical methods that we demonstrate enable quantitative, broadly applicable, and inexpensive sensing with flexibility based on a wide variety of important electroanalytical techniques (chronoamperometry, cyclic voltammetry, differential pulse voltammetry, square wave voltammetry, and potentiometry), each with different uses. Four applications demonstrate the analytical performance of the device: these involve the detection of (i) glucose in the blood for personal health, (ii) trace heavy metals (lead, cadmium, and zinc) in water for in-field environmental monitoring, (iii) sodium in urine for clinical analysis, and (iv) a malarial antigen (Plasmodium falciparum histidine-rich protein 2) for clinical research. The combination of these electrochemical capabilities in an affordable, handheld format that is compatible with any mobile phone or network worldwide guarantees that sophisticated diagnostic testing can be performed by users with a broad spectrum of needs, resources, and levels of technical expertise.

Susceptibility of Soil Bound Mercury to Gaseous Emission As a Function of Source Depth: An Enriched Isotope Tracer Investigation.

Soil mercury (Hg) emissions are an important component of the global Hg cycle. Sunlight induced photoreduction of oxidized Hg to gaseous elemental Hg is an important mechanism controlling emissions from the soil surface, however we currently understand little about how subsurface Hg stores participate in gaseous Hg cycling. Our study objective was to investigate the ability of Hg at deeper soil depths to participate in emissions. Soil fluxes were measured under controlled laboratory conditions utilizing an enriched stable Hg isotope tracer buried at 0, 1, 2, and 5 cm below the surface. Under dry and low-light conditions, the Hg isotope tracer buried at the different depths participated similarly in surface emissions (median flux: 7.5 ng m(-2) h(-1)). When the soils were wetted, Hg isotope tracer emissions increased significantly (up to 285 ng m(-2) h(-1)), with the highest fluxes (76% of emissions) originating from the surface 1 cm amended soils and decreasing with depth. Mercury associated with sandy soil up to 6 cm below the surface can be emitted, clearly demonstrating that volatilization can occur via processes unrelated to sunlight. These results have important implications for considering how long older, legacy soil Hg contamination continues to cycle between soil and atmosphere.

Paper-based potentiometric ion sensing.

This paper describes the design and fabrication of ion-sensing electrochemical paper-based analytical devices (EPADs) in which a miniaturized paper reference electrode is integrated with a small ion-selective paper electrode (ISPE) for potentiometric measurements. Ion-sensing EPADs use printed wax barriers to define electrochemical sample and reference zones. Single-layer EPADs for sensing of chloride ions include wax-defined sample and reference zones that each incorporate a Ag/AgCl electrode. In EPADs developed for other electrolytes (potassium, sodium, and calcium ions), a PVC-based ion-selective membrane is added to separate the sample zone from a paper indicator electrode. After the addition of a small volume (less than 10 µL) of sample and reference solutions to different zones, ion-sensing EPADs exhibit a linear response, over 3 orders of magnitude, in ranges of electrolyte concentrations that are relevant to a variety of applications, with a slope close to the theoretical value (59.2/z mV). Ion-selective EPADs provide a portable, inexpensive, and disposable way of measuring concentrations of electrolyte ions in aqueous solutions.

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.

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.