Optical coherence tomography-guided Brillouin microscopy highlights regional tissue stiffness differences during anterior neural tube closure in the Mthfd1l murine mutant.

Neurulation is a highly synchronized biomechanical process leading to the formation of the brain and spinal cord, and its failure leads to neural tube defects (NTDs). Although we are rapidly learning the genetic mechanisms underlying NTDs, the biomechanical aspects are largely unknown. To understand the correlation between NTDs and tissue stiffness during neural tube closure (NTC), we imaged an NTD murine model using optical coherence tomography (OCT), Brillouin microscopy and confocal fluorescence microscopy. Here, we associate structural information from OCT with local stiffness from the Brillouin signal of embryos undergoing neurulation. The stiffness of neuroepithelial tissues in Mthfd1l null embryos was significantly lower than that of wild-type embryos. Additionally, exogenous formate supplementation improved tissue stiffness and gross embryonic morphology in nullizygous and heterozygous embryos. Our results demonstrate the significance of proper tissue stiffness in normal NTC and pave the way for future studies on the mechanobiology of normal and abnormal embryonic development.

The Hip-spine Assessment of a Novel Surgical Planning Software Provides Acetabular Component Targets That Are Reliable and in Agreement With Current Clinical Recommendations.

Background: Spinopelvic immobility has been reported to increase dislocation risk following total hip arthroplasty. Surgically placing acetabular components in a functional orientation has been shown to mitigate risk. The aim of this study was to evaluate the validity and reliability of novel surgical planning software to generate clinically recommended cup targets. Methods: Hip-spine assessments were performed retrospectively on 40 patients. Five reviewers, including 3 arthroplasty-trained surgical fellows and 2 clinical research scientists performed the assessments. Hip-spine assessments consisted of measuring anterior pelvic plane tilt, sacral slope, pelvic incidence, and lumbar lordosis on standing anteroposterior pelvis and lateral standing and seated hip-spine images. Generated cup targets and a control group (40°/20° relative to the anterior pelvic plane) were compared to clinically recommended cup targets. Agreement was defined as a cup position within the recommended range or within 3° of a specific target (eg, 40° inclination) when no range was provided. Intraclass correlation coefficients were used to assess interrater and intrarater reliability, and McNemar's chi-square test was used to measure success relative to the control group. Results: The intraclass correlation coefficient was 0.88 for delta sacral slope and 0.92 for pelvic incidence-lumbar lordosis mismatch. For patients with spinopelvic risk factors, the generated targets matched the clinical recommendations in 81% of patients compared to only 16% in the control group. Conclusions: Excellent interrater and intrarater reliability was achieved using the novel surgical planning software. The resultant target values agreed with clinical recommendations to a greater extent than the control group.

Performance evaluation and model of spacesuit cooling by hydrophobic hollow fiber-membrane based water evaporation through pores.

A comprehensive mathematical model is presented that accurately estimates and predicts failure modes through the computations of heat rejection, temperature drop and lumen side pressure drop of the hollow fiber (HF) membrane-based NASA Spacesuit Water Membrane Evaporator (SWME). The model is based on mass and energy balances in terms of the physical properties of water and membrane transport properties. The mass flux of water vapor through the pores is calculated based on Knudsen diffusion with a membrane structure parameter that accounts for effective mean pore diameter, porosity, thickness, and tortuosity. Lumen-side convective heat transfer coefficients are calculated from laminar flow boundary layer theory using the Nusselt correlation. Lumen side pressure drop is estimated using the Hagen-Poiseuille equation. The coupled ordinary differential equations for mass flow rate, water temperature and lumen side pressure are solved simultaneously with the equations for mass flux and convective heat transfer to determine overall heat rejection, water temperature and lumen side pressure drop. A sensitivity analysis is performed to quantify the effect of input variability on SWME response and identify critical failure modes. The analysis includes the potential effect of organic and/or inorganic contaminants and foulants, partial pore entry due to hydrophilization, and other unexpected operational failures such as bursting or fiber damage. The model can be applied to other hollow fiber membrane-based applications such as low temperature separation and concentration of valuable biomolecules from solution.

Endemic parkinsonism: clusters, biology and clinical features.

The term 'endemic parkinsonism' refers to diseases that manifest with a dominant parkinsonian syndrome, which can be typical or atypical, and are present only in a particular geographically defined location or population. Ten phenotypes of endemic parkinsonism are currently known: three in the Western Pacific region; two in the Asian-Oceanic region; one in the Caribbean islands of Guadeloupe and Martinique; and four in Europe. Some of these disease entities seem to be disappearing over time and therefore are probably triggered by unique environmental factors. By contrast, other types persist because they are exclusively genetically determined. Given the geographical clustering and potential overlap in biological and clinical features of these exceptionally interesting diseases, this Review provides a historical reference text and offers current perspectives on each of the 10 phenotypes of endemic parkinsonism. Knowledge obtained from the study of these disease entities supports the hypothesis that both genetic and environmental factors contribute to the development of neurodegenerative diseases, not only in endemic parkinsonism but also in general. At the same time, this understanding suggests useful directions for further research in this area.

Republication of "Osteochondral Lesions of the Talus: Current Concepts in Diagnosis and Treatment".

Osteochondral lesions of the talus (OLTs) are a difficult pathologic entity to treat. They require a strong plan. Lesion size, location, chronicity, and characteristics such as displacement and the presence of subchondral cysts help dictate the appropriate treatment required to achieve a satisfactory result. In general, operative treatment is reserved for patients with displaced OLTs or for patients who have failed nonoperative treatment for 3 to 6 months. Operative treatments can be broken down into cartilage repair, replacement, and regenerative strategies. There are many promising treatment options, and research is needed to elucidate which are superior to minimize the morbidity from OLTs.

Evaluation of Mechanically-Assisted Crevice Corrosion of Different Modular Dual Mobility Constructs.

BACKGROUND: Modular dual mobility (MDM) acetabular component use is rising in total hip arthroplasty. However, concern of mechanically assisted crevice corrosion (MACC) at the shell-liner interface remains. We investigated shell-liner corrosion using retrieval analyses and corrosion chamber testing. METHODS: We analyzed fretting and corrosion on 10 matched pairs of 2 commercial MDM constructs (MDM1 and MDM2). Also, pristine pairs of Ti6Al4V shells and CoCrMo liners from 3 commercial dual mobility systems (MDM1, MDM2, and MDM3) were tested in vitro to model MACC performance. Three pairs of each were placed into an electrochemical chamber with stepwise increasing cyclic compression loads while measuring currents generated at the shell-liner taper. Onset fretting loads and fretting currents were calculated. RESULTS: Corrosion damage scores on retrieved components were low but higher in the MDM2 to MDM1 liners (P = .006), specifically outside the taper region (P = .00003). Fretting currents were higher in the MDM2 than in MDM1 or MDM3 (P = .011). Onset loads were also higher in the MDM2 (P = .001). CONCLUSION: Among retrieved liners, MDM2 tapers seem prone to non-mechanical corrosion modes. Higher onset loads and fretting currents in MDM2 tapers indicate greater MACC resistance but higher severity once corrosion begins. Differences among the devices were likely due to taper design and surface finish. Currents in all 3 were <5 µA, much lower than those observed with head-neck tapers. Our findings suggest that, among the types of corrosion observed in these MDM designs, mechanically driven corrosion may not be the most significant.

Deep Learning Phenotype Automation and Cohort Analyses of 1,946 Knees Using the Coronal Plane Alignment of the Knee Classification.

BACKGROUND: The Coronal Plane Alignment of the Knee (CPAK) classification allows for knee phenotyping which can be used in preoperative planning prior to total knee arthroplasty. We used deep learning (DL) to automate knee phenotyping and analyzed CPAK distributions in a large patient cohort. METHODS: Patients who had full-limb radiographs from a large arthritis database were retrospectively included. A DL algorithm was developed to automate CPAK knee alignment parameters including the lateral distal femoral, medial proximal tibia, hip-knee-ankle, and joint line obliquity angles. The algorithm was validated against a fellowship-trained arthroplasty surgeon. After applying the algorithm in a large patient cohort (n = 1,946 knees), the distribution of CPAK was compared across patient sex and baseline Kellgren-Lawrence (KL) scores. RESULTS: There was no significant difference in the CPAK angles (n = 140, P = .66-.98, inter-class correlation coefficient = 0.89-0.91) or phenotype classifications made by the algorithm and surgeon (P = .96). The deep learning algorithm measured the entire cohort (n = 1,946 knees, mean age 61 years [range, 46 to 80 years], 51% women) in < 5 hours. Women had more valgus CPAK phenotypes than men (P < .05). Patients who had higher KL grades at baseline (2 to 4) were more varus using the CPAK classification compared to lower KL grades (0 to 1) (P < .05). CONCLUSION: We applied an accurate, automated DL algorithm on a large patient cohort to determine knee phenotypes, helping to validate and strengthen the CPAK classification system. Analyses revealed that sex-specific and major bone loss adjustments may need to be accounted for when using this system.

In vivo tracking of KCC2b expression during early brain development.

The neuronal chloride (Cl-) exporter, KCC2, regulates neuron excitability and development and undergoes a stereotypical pattern of delayed upregulation as neurons mature. KCC2 upregulation favors neural inhibition by establishing a negative Cl- gradient, ensuring GABA-induced Cl- currents are inward and inhibitory. We developed a zebrafish fluorescent reporter line, KCC2b:mCitrine, to track KCC2 expression in vivo during early brain development. KCC2b:mCitrine was first detected at 16 h postfertilization and by day 6 labeled most central and peripheral neurons and processes. At 20 h, expression was greatest in the soma-dense basal neuroepithelium but largely absent in apical and mantle zones where differentiation and migration primarily occur, and time lapse imaging at this stage supports a postmigration upregulation of KCC2b. Central dopamine neurons showed low KCC2b expression as observed in other species. KCC2b:mCitrine fluorescence was stable over minutes in most neurons, but brightness transients observed in single cells fit our expectation for real-time tracking of KCC2b upregulation in new neurons. To further assess whether fluorescence brightness tracks KCC2b expression, zebrafish embryos were exposed to bisphenol-A (BPA), which is known to suppress KCC2 expression. Fluorescence decreased after 6 days of BPA exposure but not after 2 or 4 days, suggesting that it is an accurate but delayed indicator of KCC2b expression. KCC2b:mCitrine zebrafish present a new method for visualizing KCC2b's complex dynamics during brain development, and potentially screening compounds aimed at modulating KCC2 expression.

Re-initiation of elective total knee arthroplasty with an adapted pathway during the 2020 COVID-19 pandemic was safe and effective.

INTRODUCTION: In June 2020 when elective total knee arthroplasty (TKA) resumed after the initial COVID-19 surge, we adapted our TKA pathway focusing on a shorter hospitalization, increased home discharge, and use of post-discharge telemedicine and telerehabilitation. The purpose of this study was to evaluate if changes in postoperative care affected early TKA outcomes. MATERIALS AND METHODS: Five hundred and fifty-four patients who underwent elective primary unilateral TKA for primary osteoarthritis between June and August 2020 (study group) were matched 1:1 for age, sex, body mass index, and Charlson comorbidity index with control patients who underwent surgery between August and November 2019. Study patients were discharged 25 h earlier on average compared to controls, more frequently on the same-day or postoperative day-1 (24.9% vs. 16.1%; p = 0.001), and more frequently home (97.3% vs. 83.8%; p < 0.001). Study patients used telemedicine (11.7% vs. 0%; p < 0.001) and telerehabilitation (19.7% vs. 2.5%; p < 0.001) at higher rates than controls. Generalized estimating equations, Mann-Whitney U, and Chi-Square tests were used to compare outcomes between groups including unscheduled office visits, ER visits, readmissions, Center for Medicare and Medicaid Services (CMS) complications, manipulation under anesthesia (MUA), and patient-reported outcomes measures (PROMs). RESULTS: Rates of emergency room visits, readmissions, CMS complications, MUA, and improvements in PROMs were similar between cohorts. Study patients experienced higher rates of unscheduled outpatient visits (9.2% vs. 4.9%; p = 0.004), predominantly due to wound complications. CONCLUSIONS: A protocol implemented during the COVID-19 pandemic that leveraged a shortened hospitalization, higher rates of home discharge, and increased use of telemedicine and telerehabilitation was safe and effective.

Outcomes and Cost Analysis of a Surgical Care Unit for Outpatient Total Joint Arthroplasties Performed at a Tertiary Academic Center.

Background: Total joint arthroplasty (TJA) has shifted toward being an outpatient surgery with advances in perioperative protocols and patient selection. Most literature on outpatient arthroplasty focuses on the surgery performed in ambulatory centers. This study sought to compare (1) short-term outcomes and (2) the total procedural cost in patients who underwent TJAs at a tertiary academic center and were discharged from the accelerated surgical care unit (SCU) vs the inpatient floor unit. Methods: This is a retrospective review of 1231 procedures (637 total knee arthroplasties and 594 total hip arthroplasties) performed between January 2020 and May 2021 at 1 tertiary academic center. The minimum required follow-up duration was 90 days. Patients were divided into 2 cohorts based on discharge location (SCU vs inpatient). Patient demographics, medical comorbidities, 90-day hospital returns, and revisions were evaluated with univariate and multivariate analyses. Of the 1231 patients, 1092 had available cost data that were analyzed by a univariate analysis. Results: Patients discharged from the SCU were younger (P < .01), with lower American Society of Anesthesiologists scores (P = .04). SCU patients trended toward fewer 90-day hospital returns and revisions; however, these results were not significant. The overall encounter cost for TJA was significantly lower in patients discharged from the SCU (P < .01). Cost remained significantly less at 30 and 90 days postoperatively (P < .01). Conclusions: The present study represents the current activity at many large tertiary American academic medical centers. Results demonstrate that a unit for accelerated discharge after TJAs in a tertiary academic center is safe and cost-effective. With proper implementation, its development will enhance arthroplasty programs at a large tertiary academic center.

Front-End Design for SiPM-Based Monolithic Neutron Double Scatter Imagers.

Neutron double scatter imaging exploits the kinematics of neutron elastic scattering to enable emission imaging of neutron sources. Due to the relatively low coincidence detection efficiency of fast neutrons in organic scintillator arrays, imaging efficiency for double scatter cameras can also be low. One method to realize significant gains in neutron coincidence detection efficiency is to develop neutron double scatter detectors which employ monolithic blocks of organic scintillator, instrumented with photosensor arrays on multiple faces to enable 3D position and multi-interaction time pickoff. Silicon photomultipliers (SiPMs) have several advantageous characteristics for this approach, including high photon detection efficiency (PDE), good single photon time resolution (SPTR), high gain that translates to single photon counting capabilities, and ability to be tiled into large arrays with high packing fraction and photosensitive area fill factor. However, they also have a tradeoff in high uncorrelated and correlated noise rates (dark counts from thermionic emissions and optical photon crosstalk generated during avalanche) which may complicate event positioning algorithms. We have evaluated the noise characteristics and SPTR of Hamamatsu S13360-6075 SiPMs with low noise, fast electronic readout for integration into a monolithic neutron scatter camera prototype. The sensors and electronic readout were implemented in a small-scale prototype detector in order to estimate expected noise performance for a monolithic neutron scatter camera and perform proof-of-concept measurements for scintillation photon counting and three-dimensional event positioning.

Embryonic Hypotaurine Levels Contribute to Strain-Dependent Susceptibility in Mouse Models of Valproate-Induced Neural Tube Defects.

Valproic acid (VPA, valproate, Depakote) is a commonly used anti-seizure medication (ASM) in the treatment of epilepsy and a variety of other neurological disorders. While VPA and other ASMs are efficacious for management of seizures, they also increase the risk for adverse pregnancy outcomes, including neural tube defects (NTDs). Thus, the utility of these drugs during pregnancy and in women of childbearing potential presents a continuing public health challenge. Elucidating the underlying genetic or metabolic risk factors for VPA-affected pregnancies may lead to development of non-teratogenic ASMs, novel prevention strategies, or more targeted methods for managing epileptic pregnancies. To address this challenge, we performed unbiased, whole embryo metabolomic screening of E8.5 mouse embryos from two inbred strains with differential susceptibility to VPA-induced NTDs. We identified metabolites of differential abundance between the two strains, both in response to VPA exposure and in the vehicle controls. Notable enriched pathways included lipid metabolism, carnitine metabolism, and several amino acid pathways, especially cysteine and methionine metabolism. There also was increased abundance of ω-oxidation products of VPA in the more NTD-sensitive strain, suggesting differential metabolism of the drug. Finally, we found significantly reduced levels of hypotaurine in the susceptible strain regardless of VPA status. Based on this information, we hypothesized that maternal supplementation with L-carnitine (400 mg/kg), coenzyme A (200 mg/kg), or hypotaurine (350 mg/kg) would reduce VPA-induced NTDs in the sensitive strain and found that administration of hypotaurine prior to VPA exposure significantly reduced the occurrence of NTDs by close to one-third compared to controls. L-carnitine and coenzyme A reduced resorption rates but did not significantly reduce NTD risk in the sensitive strain. These results suggest that genetic variants or environmental exposures influencing embryonic hypotaurine status may be factors in determining risk for adverse pregnancy outcomes when managing the health care needs of pregnant women exposed to VPA or other ASMs.

Treatment Outcomes in Spinal Metastatic Disease With Indeterminate Stability.

STUDY DESIGN: Retrospective cohort study. OBJECTIVE: The purpose of this study was to compare outcomes between different treatment modalities for metastatic disease with indeterminate instability (Spinal Instability Neoplastic Score [SINS] 7-12). METHODS: We retrospectively reviewed neurologically intact patients treated for spinal metastatic disease with a SINS of 7 to 12. The cohort was stratified by treatment approach: external beam radiation therapy alone (EBRT), surgery + EBRT (S+E), and cement augmentation + EBRT (K+E). Kaplan-Meier analysis was used to assess differences in length of survival (LOS) and ability to ambulate at time of death. Multivariate analysis was performed to assess adjusted LOS and ability to ambulate at time of death. RESULTS: The cohort included 211 patients, S+E (n = 57), EBRT (n = 128), and K+E (n = 27). In the S+E group, the median LOS was 430 days, which was statistically longer than the median LOS for the EBRT group (121 days) and the K+E group (169 days). In the S+E group, 52 patients (91.2%) and in the K+E group 24 patients (92.3%) retained the ability to ambulate at their time of death compared to 99 patients (77.3%) of the EBRT patients (P = .01). The overall rate of revision treatment at the spinal level initially treated was 17.5%, S+E (15.8%), EBRT (20.3%), and K+E (7.7%). CONCLUSIONS: The length of survival, ability to maintain ambulatory ability, and revision treatment rates were all improved following surgical management and radiation therapy compared to radiation therapy alone. The authors' conclusion from these results are that patients with indeterminate spinal instability should be discussed in a multidisciplinary setting for the need of spinal stabilization in addition to radiation therapy.

Hypermethylation of PI3K-AKT signalling pathway genes is associated with human neural tube defects.

Neural tube defects (NTDs) are a group of common and severe congenital malformations. The PI3K-AKT signalling pathway plays a crucial role in the neural tube development. There is limited evidence concerning any possible association between aberrant methylation in PI3K-AKT signalling pathway genes and NTDs. Therefore, we aimed to investigate potential associations between aberrant methylation of PI3K-AKT pathway genes and NTDs. Methylation studies of PI3K-AKT pathway genes utilizing microarray genome-methylation data derived from neural tissues of ten NTD cases and eight non-malformed controls were performed. Targeted DNA methylation analysis was subsequently performed in an independent cohort of 73 NTD cases and 32 controls to validate the methylation levels of identified genes. siRNAs were used to pull-down the target genes in human embryonic stem cells (hESCs) to examine the effects of the aberrant expression of target genes on neural cells. As a result, 321 differentially hypermethylated CpG sites in the promoter regions of 30 PI3K-AKT pathway genes were identified in the microarray data. In target methylation analysis, CHRM1, FGF19, and ITGA7 were confirmed to be significantly hypermethylated in NTD cases and were associated with increased risk for NTDs. The down-regulation of FGF19, CHRM1, and ITGA7 impaired the formation of rosette-like cell aggregates. The down-regulation of those three genes affected the expression of PAX6, SOX2 and MAP2, implying their influence on the differentiation of neural cells. This study for the first time reported that hypermethylation of PI3K-AKT pathway genes such as CHRM1, FGF19, and ITGA7 is associated with human NTDs.

Driving under the influence: a multi-center evaluation of vehicular crashes in the era of cannabis legalization.

BACKGROUND: Eleven states have instituted laws allowing recreational cannabis use leading to growing public health concerns surrounding the effects of cannabis intoxication on driving safety. We hypothesized that after the 2016 legalization of cannabis in California, the use among vehicular injury patients would increase and be associated with increased injury severity. METHODS: San Diego County's five adult trauma center registries in were queried from January 2010 to June 2018 for motor vehicle or motorcycle crash patients with completed toxicology screens. Patients were stratified as toxicology negative (TOX-), positive for only THC (THC+), only blood alcohol >0.08% (ETOH+), THC+ETOH, or THC+ with any combination with methamphetamine or cocaine (M/C). County medical examiner data were reviewed to characterize THC use in those with deaths at the scene of injury. RESULTS: Of the 11,491 patients identified, there were 61.6% TOX-, 11.7% THC+, 13.7% ETOH+, 5.0% THC+ETOH, and 7.9% M/C. THC+ increased from 7.3% to 14.8% over the study period and peaked at 14.9% post-legalization in 2017. Compared with TOX- patients, THC+ patients were more likely to be male and younger. THC+ patients were also less likely to wear seatbelts (8.5% vs 14.3%, p<0.001) and had increased mean Injury Severity Score (8.4±9.4 vs 9.0±9.9, p<0.001) when compared with TOX- patients. There was no difference in in-hospital mortality between groups. From the medical examiner data of the 777 deaths on scene, 27% were THC+. DISCUSSION: THC+ toxicology screens in vehicular injury patients peaked after the 2016 legalization of cannabis. Public education on the risks of driving under the influence of cannabis should be a component of injury prevention initiatives. LEVEL OF EVIDENCE: III, Prognostic.

Unraveling the complex genetics of neural tube defects: From biological models to human genomics and back.

Neural tube defects (NTDs) are a classic example of preventable birth defects for which there is a proven-effective intervention, folic acid (FA); however, further methods of prevention remain unrealized. In the decades following implementation of FA nutritional fortification programs throughout at least 87 nations, it has become apparent that not all NTDs can be prevented by FA. In the United States, FA fortification only reduced NTD rates by 28-35% (Williams et al., 2015). As such, it is imperative that further work is performed to understand the risk factors associated with NTDs and their underlying mechanisms so that alternative prevention strategies can be developed. However, this is complicated by the sheer number of genes associated with neural tube development, the heterogeneity of observable phenotypes in human cases, the rareness of the disease, and the myriad of environmental factors associated with NTD risk. Given the complex genetic architecture underlying NTD pathology and the way in which that architecture interacts dynamically with environmental factors, further prevention initiatives will undoubtedly require precision medicine strategies that utilize the power of human genomics and modern tools for assessing genetic risk factors. Herein, we review recent advances in genomic strategies for discovering genetic variants associated with these defects, and new ways in which biological models, such as mice and cell culture-derived organoids, are leveraged to assess mechanistic functionality, the way these variants interact with other genetic or environmental factors, and their ultimate contribution to human NTD risk.

Producing Vaccines against Enveloped Viruses in Plants: Making the Impossible, Difficult.

The past 30 years have seen the growth of plant molecular farming as an approach to the production of recombinant proteins for pharmaceutical and biotechnological uses. Much of this effort has focused on producing vaccine candidates against viral diseases, including those caused by enveloped viruses. These represent a particular challenge given the difficulties associated with expressing and purifying membrane-bound proteins and achieving correct assembly. Despite this, there have been notable successes both from a biochemical and a clinical perspective, with a number of clinical trials showing great promise. This review will explore the history and current status of plant-produced vaccine candidates against enveloped viruses to date, with a particular focus on virus-like particles (VLPs), which mimic authentic virus structures but do not contain infectious genetic material.