Mayo Clinic VT calculator: A practical tool for accurate wide complex tachycardia differentiation.

The discrimination of ventricular tachycardia (VT) versus supraventricular wide complex tachycardia (SWCT) via 12-lead electrocardiogram (ECG) is crucial for achieving appropriate, high-quality, and cost-effective care in patients presenting with wide QRS complex tachycardia (WCT). Decades of rigorous research have brought forth an expanding arsenal of applicable manual algorithm methods for differentiating WCTs. However, these algorithms are limited by their heavy reliance on the ECG interpreter for their proper execution. Herein, we introduce the Mayo Clinic ventricular tachycardia calculator (MC-VTcalc) as a novel generalizable, accurate, and easy-to-use means to estimate VT probability independent of ECG interpreter competency. The MC-VTcalc, through the use of web-based and mobile device platforms, only requires the entry of computerized measurements (i.e., QRS duration, QRS axis, and T-wave axis) that are routinely displayed on standard 12-lead ECG recordings.

The history of orthopaedic use of nitinol compression staples.

INTRODUCTION: The use of nitinol continuous compression staples has shown clinical utility in the management of various orthopaedic injuries. While literature is most robust in the realm of foot/ankle and spine surgery, the use of nitinol staples has been documented in fixation of wrist, olecranon, patella, and pelvis fractures. METHODOLOGY: A narrative review was conducted by searching three online databases - PubMed, Web of Science, and Cochrane using the terms "Nitinol" and "Staple" published between 2003 and 2023. A total of 42 articles met inclusion/exclusion criteria and were included in this review. REVIEW: Literature outside of foot/ankle and spine surgery is largely limited to biomechanical studies, case reports, and finite element analyses. The literature is summarized within this review by anatomic location including foot/ankle, lower extremity, hand, upper extremity, spine, and pelvis. CONCLUSION: Existing literature demonstrates a diverse array of applications for nitinol continuous compression staples in both axial and appendicular orthopaedic care. Advantages of these implants include ease of application, ability to capture small bony fragments, continuous compression across a fracture or arthrodesis, and full coaptation which maximizes the surface area for healing and/or fusion.

A strategy for high antibody expression with low anti-drug antibodies using AAV9 vectors.

Introduction: Use of adeno-associated virus (AAV) vectors is complicated by host immune responses that can limit transgene expression. Recent clinical trials using AAV vectors to deliver HIV broadly neutralizing antibodies (bNAbs) by intramuscular administration resulted in poor expression with anti-drug antibodies (ADA) responses against the bNAb. Methods: Here we compared the expression of, and ADA responses against, an anti-SIV antibody ITS01 when delivered by five different AAV capsids. We first evaluated ITS01 expression from AAV vectors three different 2A peptides. Rhesus macaques were selected for the study based on preexisiting neutralizing antibodies by evaluating serum samples in a neutralization assay against the five capsids used in the study. Macaques were intramuscularly administered AAV vectors at a 2.5x10^12 vg/kg over eight administration sites. ITS01 concentrations and anti-drug antibodies (ADA) were measured by ELISA and a neutralization assay was conducted to confirm ex vivo antibody potency. Results: We observed that ITS01 expressed three-fold more efficiently in mice from AAV vectors in which heavy and light-chain genes were separated by a P2A ribosomal skipping peptide, compared with those bearing F2A or T2A peptides. We then measured the preexisting neutralizing antibody responses against three traditional AAV capsids in 360 rhesus macaques and observed that 8%, 16%, and 42% were seronegative for AAV1, AAV8, and AAV9, respectively. Finally, we compared ITS01 expression in seronegative macaques intramuscularly transduced with AAV1, AAV8, or AAV9, or with the synthetic capsids AAV-NP22 or AAV-KP1. We observed at 30 weeks after administration that AAV9- and AAV1-delivered vectors expressed the highest concentrations of ITS01 (224 µg/mL, n=5, and 216 µg/mL, n=3, respectively). The remaining groups expressed an average of 35-73 µg/mL. Notably, ADA responses against ITS01 were observed in six of the 19 animals. Lastly, we demonstrated that the expressed ITS01 retained its neutralizing activity with nearly the same potency of purified recombinant protein. Discussion: Overall, these data suggest that the AAV9 capsid is a suitable choice for intramuscular expression of antibodies in nonhuman primates.

A lentiviral vector B cell gene therapy platform for the delivery of the anti-HIV-1 eCD4-Ig-knob-in-hole-reversed immunoadhesin.

Barriers to effective gene therapy for many diseases include the number of modified target cells required to achieve therapeutic outcomes and host immune responses to expressed therapeutic proteins. As long-lived cells specialized for protein secretion, antibody-secreting B cells are an attractive target for foreign protein expression in blood and tissue. To neutralize HIV-1, we developed a lentiviral vector (LV) gene therapy platform for delivery of the anti-HIV-1 immunoadhesin, eCD4-Ig, to B cells. The EµB29 enhancer/promoter in the LV limited gene expression in non-B cell lineages. By engineering a knob-in-hole-reversed (KiHR) modification in the CH3-Fc eCD4-Ig domain, we reduced interactions between eCD4-Ig and endogenous B cell immunoglobulin G proteins, which improved HIV-1 neutralization potency. Unlike previous approaches in non-lymphoid cells, eCD4-Ig-KiHR produced in B cells promoted HIV-1 neutralizing protection without requiring exogenous TPST2, a tyrosine sulfation enzyme required for eCD4-Ig-KiHR function. This finding indicated that B cell machinery is well suited to produce therapeutic proteins. Lastly, to overcome the inefficient transduction efficiency associated with VSV-G LV delivery to primary B cells, an optimized measles pseudotyped LV packaging methodology achieved up to 75% transduction efficiency. Overall, our findings support the utility of B cell gene therapy platforms for therapeutic protein delivery.

Curvafix: A novel implant for pelvic fragility fractures.

Fragility fractures of the pelvis are a frequently encountered injury in orthopedic practice. These injuries occur in patients with diminished bone density secondary to low energy mechanisms. This injury is associated with increased mortality and decreased quality of life. These fractures have been treated conservatively and with open or percutaneous surgical fixation. This case report illustrates the use of the Curvafix, a novel intramedullary implant used to treat a patient with a pelvic fragility fracture.

Seroprevalence of Adeno-Associated Virus Neutralizing Antibodies in Males with Duchenne Muscular Dystrophy.

Adeno-associated virus (AAV)-based gene therapies are emerging strategies in Duchenne muscular dystrophy (DMD) treatment. Exposure to wild-type AAV can lead to development of neutralizing antibodies (NAbs) and blocking of AAV transduction, thereby limiting the delivery of AAV vector-based gene therapy. Therefore, it is imperative to check for the presence of AAV NAbs in a patient who is a candidate for gene therapy. We prospectively enrolled 101 genetically confirmed males with DMD (median age 11 years, 48% ambulatory and 59% on steroids) and performed AAV neutralization assays against AAV2, AAV8, AAV9, and AAVrh74 serotypes. The serotype analysis showed that AAV9 (36%) and AAVrh74 (32%) seroprevalence was lower compared with AAV2 (56%) and AAV8 (47%). Interestingly, age was not correlated with NAb titer for any of the capsids. NAb responses were observed at a higher frequency in African American participants and at a lower frequency in Caucasian participants for all four serotypes. Further analysis showed no significant differences in NAb titers regardless of serotype and whether participants were taking steroids or not. Finally, we observed higher AAV8, AAV9, and AAVrh74 seroprevalence and significantly higher AAV2 and AAV8 NAb titers in participants who were ambulatory compared with nonambulatory participants. Overall, these data identify AAV9 and AAVrh74 as the two serotypes with lower pre-existing NAb titers in this study's cohort of 101 males with DMD, possibly showing their utility in future gene therapy applications in treatment of this cohort of patients with DMD.

The role of the lateral orbitofrontal cortex in creating cognitive maps.

We use mental models of the world-cognitive maps-to guide behavior. The lateral orbitofrontal cortex (lOFC) is typically thought to support behavior by deploying these maps to simulate outcomes, but recent evidence suggests that it may instead support behavior by underlying map creation. We tested between these two alternatives using outcome-specific devaluation and a high-potency chemogenetic approach. Selectively inactivating lOFC principal neurons when male rats learned distinct cue-outcome associations, but before outcome devaluation, disrupted subsequent inference, confirming a role for the lOFC in creating new maps. However, lOFC inactivation surprisingly led to generalized devaluation, a result that is inconsistent with a complete mapping failure. Using a reinforcement learning framework, we show that this effect is best explained by a circumscribed deficit in credit assignment precision during map construction, suggesting that the lOFC has a selective role in defining the specificity of associations that comprise cognitive maps.

Fast Protocols for Characterizing Antibody-Peptide Binding.

Microarray assay formats gained popularity in the 1990s, first implemented in DNA-based arrays but later adopted for use with proteins, namely antibodies, peptides, low molecular weight (LMW) molecules, such as lipids, and even tissues. In nucleic acid-based affinity assays and arrays, but not in protein or peptide arrays, the specificity and affinity of complementary strand interactions can be deduced from or adjusted through modifications to the nucleotide sequence. Arrays of LMW molecules are characterized by largely uniform but low binding affinities. Multiplexed protein-based affinity assays, such as microarrays, might present an additional challenge due to heterogeneity of antigen properties and of their binding affinities. The use of peptides instead of proteins reduces physical heterogeneity of these reagents through either the widened peptide selection options or rational sequence engineering. However, rational engineering of binding affinities remains an unmet need, and peptide-binding affinities to the respective antipeptide antibodies could vary by orders of magnitude. Hence, multiplexing of such assays by using a microarray format and data analysis and interpretation requires some knowledge of their binding affinities. Low-throughput binding assays to characterize such peptide-antipeptide antibodies interactions are widely available, but scaling-up of traditional protein- and peptide-binding assays might present practical challenges. Here, we describe fast label-free practical approach especially suitable for estimating peptide-binding affinities. The method in question relies on commercially available biolayer interferometry-based equipment with a protocol which can be easily scaled-up, subject to user needs and equipment availability.

Peptides and Anti-peptide Antibodies for Small- and Medium-Scale Peptide and Anti-peptide Affinity Microarrays.

This chapter describes the principles for selection of antigenic peptides for the development of anti-peptide antibodies suitable for microarray-based multiplex affinity assays and optional mass spectrometry detection. The methods described here are mostly applicable to small- and medium-scale multiplex affinity assay and microarrays. Although the same principles of peptide selection may also be applied to larger-scale arrays (with 100+ features), informatics software and printing methods may well differ. Due to the sheer number of proteins/peptides to be processed and analyzed, dedicated software with high processing capacity and enterprise-level array robotics may be required for larger-scale efforts. This report aims to provide practical advice to those seeking to develop or use arrays with up to ~100 different peptide or protein features.

Calcium activity is a degraded estimate of spikes.

Recording action potentials extracellularly during behavior has led to fundamental discoveries regarding neural function-hippocampal neurons respond to locations in space,1 motor cortex neurons encode movement direction,2 and dopamine neurons signal reward prediction errors3-observations undergirding current theories of cognition,4 movement,5 and learning.6 Recently it has become possible to measure calcium flux, an internal cellular signal related to spiking. The ability to image calcium flux in anatomically7,8 or genetically9 identified neurons can extend our knowledge of neural circuit function by allowing activity to be monitored in specific cell types or projections, or in the same neurons across many days. However, while initial studies were grounded in prior unit recording work, it has become fashionable to assume that calcium is identical to spiking, even though the spike-to-fluorescence transformation is nonlinear, noisy, and unpredictable under real-world conditions.10 It remains an open question whether calcium provides a high-fidelity representation of single-unit activity in awake, behaving subjects. Here, we have addressed this question by recording both signals in the lateral orbitofrontal cortex (OFC) of rats during olfactory discrimination learning. Activity in the OFC during olfactory learning has been well-studied in humans,11,12,13,14 nonhuman primates,15,16 and rats,17,18,19,20,21 where it has been shown to signal information about both the sensory properties of odor cues and the rewards they predict. Our single-unit results replicated prior findings, whereas the calcium signal provided only a degraded estimate of the information available in the single-unit spiking, reflecting primarily reward value.

Technical considerations for the management of segmental osseous defects with an internal bone transport nail.

INTRODUCTION: Traditionally, distraction osteogenesis has been accomplished with an external fixator. All internal transport utilizing magnetic intramedullary nails is a newer technique for bone reconstruction. The Precice Bone Transport Nail is a new implant that allows for noninvasive transport via a magnetically driven motor. AREAS COVERED: This report describes the function of the Bone Transport Nail along with the technical considerations on how to successfully manage bone defects with this new technology. Appropriate use of the nail, preoperative planning, intraoperative considerations, and postoperative management are discussed in detail. EXPERT OPINION: The Precice Bone Transport Nail utilizes the technology of the original Precice nail to provide an all-internal option for reconstruction of intercalary defects. This obviates the need for an additional plate with a standard Precice nail when performing bone transport and allows for a less invasive option that decreases operating room time. It provides a more cosmetic result than external fixation and avoids the risks of pin tract infection. Preoperative planning is essential to appropriate execution of the operative procedure and to perform a successful transport. A thorough understanding of the nail design and limitations are a prerequisite as this implant is significantly different from a standard intramedullary nail.

High concordance of ELISA and neutralization assays allows for the detection of antibodies to individual AAV serotypes.

Prescreening of participants in clinical trials that use adeno-associated virus (AAV) vectors is required to identify naive participants, as preexisting neutralizing antibodies can limit the efficacy of AAV gene therapies. The presence of antibodies to individual AAV serotypes is typically detected by neutralization assay. To streamline the screening process, we compared an ELISA-based screening method with a neutralization assay for the detection of antibodies against AAV1, AAV8, and AAV9 in a collection of 50 rhesus macaque sera and 20 human sera. We observed a high level of concordance between the two assays (Pearson r > 0.8) for all three serotypes in both sample sets. We thus investigated pre- vs post-vector inoculation sera samples from rhesus macaques that received AAV1 or AAV8 vector inoculations for cross-reactive anti-AAV antibodies. All 12 macaques seroconverted to the vector they received, but many also reacted to the other serotypes. Our results validate an easy-to-use ELISA for reliable detection of antibodies to individual serotypes of AAV. Our results also demonstrate that an antibody response post-AAV inoculation may partially cross-react with other AAV serotypes. Overall, these results suggest that either assay can be used by academic labs for prescreening samples for preexisting anti-AAV antibodies.

Estimation of the in vivo neutralization potency of eCD4Ig and conditions for AAV-mediated production for SHIV long-term remission.

The engineered protein eCD4Ig has emerged as a promising approach to achieve HIV remission in the absence of antiviral therapy. eCD4Ig neutralizes nearly all HIV-1 isolates and induces antibody-dependent cell-mediated cytotoxicity (ADCC) in vitro. To characterize the in vivo antiviral neutralization and possible ADCC effects of eCD4Ig, we fit mathematical models to eCD4Ig, anti­eCD4Ig-drug antibody (ADA), and viral load kinetics from healthy and simian-human immunodeficiency virus AD8 (SHIV-AD8) infected nonhuman primates that were treated with single or sequentially dosed eCD4Ig passive administrations. Our model predicts that eCD4Ig transiently decreases SHIV viral loads due to neutralization only with an in vivo IC50 of ~25 µg/ml but with limited effect due to ADA. Simulations suggest that endogenous, continuous expression of eCD4Ig at levels greater than 105 µg/day, as is possible with Adeno-associated virus (AAV) vector-based production, could overcome the diminishing effects of ADA and allow for long-term remission of SHIV viremia in nonhuman primates.

Anterior cingulate neurons signal neutral cue pairings during sensory preconditioning.

Of all frontocortical subregions, the anterior cingulate cortex (ACC) has perhaps the most overlapping theories of function.1-3 Recording studies in rats, humans, and other primates have reported diverse neural responses that support many theories,4-12 yet nearly all these studies have in common tasks in which one event reliably predicts another. This leaves open the possibility that ACC represents associative pairing of events, independent of their overt biological significance. Sensory preconditioning13 provides an opportunity to test this. In the first phase, preconditioning, value-neutral sensory stimuli are paired (A→B). To test whether this was learned, subjects are given standard conditioning during which one of the previously neutral sensory cues is paired with a biologically meaningful outcome (B→outcome). During the final probe test, the neutral cue which was never paired with a biologically meaningful outcome is presented alone (A→) and will elicit a conditional response, suggesting that subjects had learned the associative structure during preconditioning and use that knowledge to infer presentation of the biologically relevant outcome (A→B→outcome). Inference-based responding demonstrates a fundamental property of model-based reasoning14,15 and requires learning of the associations between neutral stimuli before rewards are introduced.16-19 ACC neurons developed firing patterns that reflected the learning of sensory associations during preconditioning, even though no rewards were present. The strength of these correlates predicted rats' ability to later mobilize and use that associative information during the probe test. These results demonstrate that clear biological significance is not necessary to produce correlates of learning in ACC.

Plate-Assisted Bone Segment Transport Versus Precice Bone Transport Nail.

SUMMARY: Management of segmental long bone defects is a complex and challenging undertaking for orthopaedic surgeons. These defects can be encountered in cases of high-energy trauma, tumor resection, or infection, and they are often associated with significant soft tissue injury. Traditional approaches of bone transport rely on external fixation and principles of distraction osteogenesis. Plate-assisted bone segment transport (PABST) using the Precice limb lengthening nail has been adapted for use in bone transport with the use of a plate in an effort to eliminate the need for external fixation and its associated complications. Recently, the arrival of the Precice Bone Transport (PBT) System intramedullary nail eliminates the need for plating and some of the problems encountered in PABST; however, it also introduces some new issues. PABST and the PBT nail have become viable alternatives to bone transport using a frame; however, each has its own unique set of advantages and disadvantages. Although the problems of using external fixation devices are eliminated with these techniques, there is less forgiveness in execution and very little chance of correcting as the transport is underway. The arrival of the PBT nail does not eliminate the need for PABST as seen by the difficulty maintaining alignment in short metaphyseal segments. This review reflects the current state of these methods based on available evidence; however, optimization of the protocol for transport using PABST and the PBT nail will require additional cases and data.

Spatial Representations in Rat Orbitofrontal Cortex.

The orbitofrontal cortex (OFC) and hippocampus share striking cognitive and functional similarities. As a result, both structures have been proposed to encode "cognitive maps" that provide useful scaffolds for planning complex behaviors. However, while this function has been exemplified by spatial coding in neurons of hippocampal regions-particularly place and grid cells-spatial representations in the OFC have been investigated far less. Here we sought to address this by recording OFC neurons from male rats engaged in an open-field foraging task like that originally developed to characterize place fields in rodent hippocampal neurons. Single-unit activity was recorded as rats searched for food pellets scattered randomly throughout a large enclosure. In some sessions, particular flavors of food occurred more frequently in particular parts of the enclosure; in others, only a single flavor was used. OFC neurons showed spatially localized firing fields in both conditions, and representations changed between flavored and unflavored foraging periods in a manner reminiscent of remapping in the hippocampus. Compared with hippocampal recordings taken under similar behavioral conditions, OFC spatial representations were less temporally reliable, and there was no significant evidence of grid tuning in OFC neurons. These data confirm that OFC neurons show spatial firing fields in a large, two-dimensional environment in a manner similar to hippocampus. Consistent with the focus of the OFC on biological meaning and goals, spatial coding was weaker than in hippocampus and influenced by outcome identity.SIGNIFICANCE STATEMENT The orbitofrontal cortex (OFC) and hippocampus have both been proposed to encode "cognitive maps" that provide useful scaffolds for planning complex behaviors. This function is exemplified by place and grid cells identified in hippocampus, the activity of which maps spatial environments. The current study directly demonstrates very similar, though not identical, spatial representatives in OFC neurons, confirming that OFC-like hippocampus-can represent a spatial map under the appropriate experimental conditions.

Prospective representations in rat orbitofrontal ensembles.

The orbitofrontal cortex (OFC) has been proposed to encode expected outcomes, which is thought to be important for outcome-directed behavior. However, such neural encoding can also often be explained by the recall of information about the recent past. To dissociate the retrospective and prospective aspects of encoding in the OFC, we designed a nonspatial, continuous, alternating odor-sequence task that mimicked a continuous T-maze. The task consisted of two alternating sequences of four odor-guided trials (2 sequences × 4 positions). In each trial, rats were asked to make a "go" or "no-go" action based on a fixed odor-reward contingency. Odors at both the first and last positions were distinct across the two sequences, such that they resembled unique paths in the past and future, respectively; odors at positions in between were the same and thus resembled a common path. We trained classifiers using neural activity to distinguish between either sequences or positions and asked whether the neural activity patterns in the common path were more like the ones in the past or the future. We found a proximal prospective code for sequence information as well as a distal perspective code for positional information, the latter of which was closely associated with rats' ability to predict future outcomes. This study demonstrates a behaviorally relevant predictive code in rat OFC. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

The orbitofrontal cartographer.

Theories of orbitofrontal cortex (OFC) function have evolved substantially over the last few decades. There is now a general consensus that the OFC is important for predicting aspects of future events and for using these predictions to guide behavior. Yet the precise content of these predictions and the degree to which OFC contributes to agency contingent upon them has become contentious, with several plausible theories advocating different answers to these questions. In this review we will focus on three of these ideas-the economic value, credit assignment, and cognitive map hypotheses-describing both their successes and failures. We will propose that these failures hint at a more nuanced and perhaps unique role for the OFC, particularly the lateral subdivision, in supporting the proposed functions when an underlying model or map of the causal structures in the environment must be constructed or updated. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Is the core function of orbitofrontal cortex to signal values or make predictions?

One dominant hypothesis about the function of the orbitofrontal cortex (OFC) is that the OFC signals the subjective values of possible outcomes to other brain areas for learning and decision making. This popular view generally neglects the fact that OFC is not necessary for simple value-based behavior (i.e., when values have been directly experienced). An alternative, emerging view suggests that OFC plays a more general role in representing structural information about the task or environment, derived from prior experience, and relevant to predicting behavioral outcomes, such as value. From this perspective, value signaling is simply one derivative of the core underlying function of OFC. New data in favor of both views have been accumulating rapidly. Here we review these new data in discussing the relative merits of these two ideas.