An assessment of the Midwest climate adaptation network: A call for improved coordination and collaboration.

Climate adaptation and the management of climate impacts require cross-sectoral and regional coordination and collaboration, but presently there is no thorough assessment of the adaptation network in the Midwest United States to evaluate how well it achieves such collaboration. We investigated the climate adaptation network across the Midwest to inform the strategic agenda for a climate adaptation boundary organization in Minnesota - the University of Minnesota Climate Adaptation Partnership (MCAP). We identified 150 organizations and more than 500 unique connections between them. About ten organizations with more than 25 connections each link the existing Midwest climate adaptation network, but most organizations have fewer than five connections. This asymmetry can affect the flow of resources such as information, technical assistance, and financial support. It can also hinder coordination and collaboration as called for by the Intergovernmental Panel on Climate Change (IPCC et al., 2019). The Midwest adaptation network is not well-balanced with respect to the adaptation cycle: many organizations focus on understanding or planning for climate change, with few organizations focused on problem identification, plan implementation, or monitoring. The gaps identified here suggest that MCAP and other regional adaptation organizations can (1) improve cross-sectoral and intraregional coordination and collaboration, and (2) fill gaps in the adaptation cycle, particularly implementation and monitoring. As more communities and jurisdictions move beyond climate planning toward adaptation implementation and management, and as an increasing number of state, federal and private sector funds become available to support implementation, climate service providers such as MCAP should evaluate their services and capacities and adapt alongside the communities they support.

Ultrasensitive Detection of Aß42 Seeds in Cerebrospinal Fluid with a Nanopipette-Based Real-Time Fast Amyloid Seeding and Translocation Assay.

In this work, early-stage Aß42 aggregates were detected using a real-time fast amyloid seeding and translocation (RT-FAST) assay. Specifically, Aß42 monomers were incubated in buffer solution with and without preformed Aß42 seeds in a quartz nanopipette coated with L-DOPA. Then, formed Aß42 aggregates were analyzed on flyby resistive pulse sensing at various incubation time points. Aß42 aggregates were detected only in the sample with Aß42 seeds after 180 min of incubation, giving an on/off readout of the presence of preformed seeds. Moreover, this RT-FAST assay could detect preformed seeds spiked in 4% cerebrospinal fluid/buffer solution. However, in this condition, the time to detect the first aggregates was increased. Analysis of Cy3-labeled Aß42 monomer adsorption on a quartz substrate after L-DOPA coating by confocal fluorescence spectroscopy and molecular dynamics simulation showed the huge influence of Aß42 adsorption on the aggregation process.

A rare case of cystic ganglionosis in a child with associated imaging findings.

Ganglia are benign soft tissue masses that are found adjacent to joints and tendons. They can be multifocal but they are rarely more numerous than a few around any given joint. "Cystic ganglionosis" has been used to describe a condition in which multifocal and extensive ganglia are present. We present a rare case of cystic ganglionosis in a Caucasian girl with clinical symptoms detected at 6 months of age. To the authors' knowledge, only a single other case report of cystic ganglionosis is documented in the English medical literature. The ganglia in this case are more extensive, manifested at an earlier age and caused erosions of multiple bones, a rarely observed complication of ganglia. Additionally, radiograph, MR and sonographic images collected over 9 years time allows for a detailed description of the imaging characteristics of this case of cystic ganglionosis, and offers unique insight into the natural history of this diagnosis. Extensive ganglia in multiple locations in a young child should alert clinicians to the possibility of cystic ganglionosis. Disease progression may lead to deleterious effects on bone warranting the use of maintenance imaging and possibly surgical resection of symptomatic lesions.

Characterizing Prion-Like Protein Aggregation: Emerging Nanopore-Based Approaches.

Prion-like protein aggregation is characteristic of numerous neurodegenerative diseases, such as Alzheimer's and Parkinson's diseases. This process involves the formation of aggregates ranging from small and potentially neurotoxic oligomers to highly structured self-propagating amyloid fibrils. Various approaches are used to study protein aggregation, but they do not always provide continuous information on the polymorphic, transient, and heterogeneous species formed. This review provides an updated state-of-the-art approach to the detection and characterization of a wide range of protein aggregates using nanopore technology. For each type of nanopore, biological, solid-state polymer, and nanopipette, discuss the main achievements for the detection of protein aggregates as well as the significant contributions to the understanding of protein aggregation and diagnostics.

Improving Naloxone Co-prescribing Through Clinical Decision Support.

BACKGROUND: Despite national guidelines recommending naloxone co-prescription with high-risk medications, rates remain low nationally. This was reflected at our institution with remarkably low naloxone prescribing rates. We sought to determine if a clinical decision support (CDS) tool could increase rates of naloxone co-prescribing with high-risk prescriptions. METHODS:  An alert in the electronic health record was triggered upon signing an order for a high-risk opioid medication without a naloxone co-prescription. We examined all opioid prescriptions written by family and general internal medicine practitioners at the University of Iowa Hospitals and Clinics in outpatient encounters between November 30, 2020, and February 28, 2022. Once triggered by a high-risk prescription, the CDS tool had the option to choose an order set with an automatically selected co-prescription for naloxone along with patient instructions automatically added to the patient's after-visit summary (AVS). We examined the monthly percentage of patients receiving Schedule II opioid prescriptions ≥90 morphine milliequivalents (MME)/day who received concurrent naloxone prescriptions in the 12 months before the CDS went live and the three months following go-live. RESULTS:  Concurrent naloxone prescriptions increased from 1.1% in the 12 months prior to implementation in November 2021 to 9.4% (p<0.001) during the post-intervention period across eight family medicine and internal medicine clinics. DISCUSSION:  This single-center quality improvement project with retrospective analysis demonstrates the potential efficacy of a single CDS tool in increasing the rate of naloxone prescription. The impact of such prescribing on overall mortality requires further research. CONCLUSIONS: The CDS tool was easy to implement and improved rates of appropriate naloxone co-prescribing.

Spatiotemporal dynamics of the proton motive force on single bacterial cells.

Electrochemical gradients across biological membranes are vital for cellular bioenergetics. In bacteria, the proton motive force (PMF) drives essential processes like adenosine triphosphate production and motility. Traditionally viewed as temporally and spatially stable, recent research reveals a dynamic PMF behavior at both single-cell and community levels. Moreover, the observed lateral segregation of respiratory complexes could suggest a spatial heterogeneity of the PMF. Using a light-activated proton pump and detecting the activity of the bacterial flagellar motor, we perturb and probe the PMF of single cells. Spatially homogeneous PMF perturbations reveal millisecond-scale temporal dynamics and an asymmetrical capacitive response. Localized perturbations show a rapid lateral PMF homogenization, faster than proton diffusion, akin to the electrotonic potential spread observed in passive neurons, explained by cable theory. These observations imply a global coupling between PMF sources and consumers along the membrane, precluding sustained PMF spatial heterogeneity but allowing for rapid temporal changes.

Successfully transitioning an interruptive alert into a non-interruptive alert for central line dressing changes in the Neonatal Intensive Care Unit.

BACKGROUND: Interruptive alerts are known to be associated with clinician alert fatigue, and poorly performing alerts should be evaluated for alternative solutions. An interruptive alert to remind clinicians about a required peripherally inserted central catheter (PICC) dressing change within the first 48-hours after placement resulted in 617 firings in a 6-month period with only 11 (1.7%) actions taken from the alert. OBJECTIVE: To enhance a poorly functioning interruptive alert by converting it to a non-interruptive alert aiming to improve compliance with the institutional PICC dressing change protocol. The primary outcome was to measure the percentage of initial PICC dressing changes that occurred beyond the recommended 48-hour timeframe after PICC placement. Secondary outcomes included measuring the time to first dressing change and, qualitatively, if this solution could replace the manual process of maintaining a physical list of patients. METHODS: A clinical informatics team met with stakeholders to evaluate the clinical workflow and identified an additional need to track which patients qualified for dressing changes. A non-interruptive patient column clinical decision support (CDS) tool was created to replace an interruptive alert. A pre-post intervention mixed-methods cohort study was conducted between January 2022 - November 2022. RESULTS: The number of patients with overdue PICC dressing changes decreased from 21.9% (40/183) to 7.8% (10/128) of eligible patients (p <0.001), and mean time to first PICC dressing changes also significantly decreased from 40.8 hours to 30.7 hours (p = 0.02). There was universal adoption of the CDS tool, and clinicians no longer used the manual patient list. CONCLUSIONS: While previous studies have reported that non-interruptive CDS may not be as effective as interruptive CDS, this case report demonstrates that developing a population-based CDS in the patient list column that provides an additional desired functionality to clinicians may result in improved adoption of CDS.

Secondary Nucleation of Aß Revealed by Single-Molecule and Computational Approaches.

Understanding the mechanisms underlying amyloid-ß (Aß) aggregation is pivotal in the context of Alzheimer's disease. This study aims to elucidate the secondary nucleation process of Aß42 peptides by combining experimental and computational methods. Using a newly developed nanopipette-based amyloid seeding and translocation assay, confocal fluorescence spectroscopy, and molecular dynamics simulations, the influence of the seed properties on Aß aggregation is investigated. Both fragmented and unfragmented seeds played distinct roles in the formation of oligomers, with fragmented seeds facilitating the formation of larger aggregates early in the incubation phase. The results show that secondary nucleation leads to the formation of oligomers of various sizes and structures as well as larger fibrils structured in ß-sheets. From these findings a mechanism of secondary nucleation involving two types of aggregate populations, one released and one growing on the mother fiber is proposed.

Aß42 fibril and non-fibril oligomers characterization using a nanopipette.

The Aß42 aggregates with different structures and morphology was investigated through a single molecule label-free technique. To this end, the quartz nanopipettes were functionalized with polyethylene glycol. The set of Aß42- epigallocatechin-3-gallate fibrils with length (from 85 nm to 250 nm) obtained by sonication was detected. The comparison of experimental and computed value of the amplitude of relative current blockade using a geometrical model show that for fibrils longer than 80 nm, the discriminating parameter is their diameter. Then, non-fibril oligomers obtain from Aß42(Osaka) aggregation at different time seed was investigated. The analysis of the amplitude of relative current blockade shows that detected oligomers are smaller than 30 nm regardless the aggregation time. In addition, the wide distributions of the dwell time suggests the polymorph character of the sample.

Combining iontronic, chromatography and nanopipette for Aß42 aggregates detection and separation.

In this work, we aim to capture, detect and analysis at single molecule level Aß42 aggregates. To this end, two strategies of track-etched nanopore membranes functionalization were investigated. The first one uses an aptamer and requires only three steps, whereas the second strategy uses Lecanemab antibodies and requires six steps. Out of the two presented strategies, the second one was found to be the most suitable to detect Aß42 aggregates using a quick current-voltage readout. The resulting single nanopore was then upscale to multipore membranes to capture the Aß42 aggregates before analysis through them through a single-molecule approach. By comparing the species present in the retentate and filtrate, we confirmed the membrane's affinity for the larger Aß42 aggregates present in the sample. We found that chromatographic membranes combined with an ionic diode for binary on/off readout are powerful tools for detecting rare biomarkers before single molecule analysis.

Cross-Section Observational Study to Assess Antimicrobial Resistance Prevalence among Bovine Respiratory Disease Bacterial Isolates from Commercial US Feedlots.

Antimicrobial resistance (AMR) is a global public health threat that jeopardizes efficacy of antibiotics in veterinary and human medicine. Antibiotics are commonly administered to target the bacterial component of bovine respiratory disease (BRD). The objectives of this study were to obtain a better understanding of antibiotic resistance in BRD-associated bacteria (Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni), investigate the clinical significance of AMR by monitoring clinical outcomes, and determine if regional differences exist in AMR trends. Deep pharyngeal swabs were used to sample beef cattle at initial BRD diagnosis (n = 453) from US feedlots representing three geographic regions. Organisms were identified by bacterial culture and subjected to broth microdilution antimicrobial susceptibility testing. Bacterium prevalence include P. multocida (36.0%), M. haemolytica (32.7%), and H. somni (28.5%). Of the Histophilus isolates, 39.5% were resistant to at least one antimicrobial, compared to 11.7% and 8.8% Pasteurella and Mannheimia, respectively. Non-susceptibility across all organisms was 5.7 X more likely in animals that received metaphylaxis, than those that did not (p < 0.0001; OR 5.7; CI 2.6-12.5). During days on feed 21-40, non-susceptibility of Histophilus was 8.7 X more likely than Mannheimia (p = 0.0002; OR 8.7; CI 2.8 to 27.4) and 6 X more likely than Pasteurella (p = 0.0016; OR 6.0; CI 2.0-18.0).

Real-Time Fast Amyloid Seeding and Translocation of α-Synuclein with a Nanopipette.

The detection to α-synuclein (αS) assemblies as a biomarker of synucleinopathies is an important challenge for further development of an early diagnosis tool. Here, we present proof of concept real-time fast amyloid seeding and translocation (RT-FAST) based on a nanopipette that combines in one unique system a reaction vessel to accelerate the seed amplification and nanopore sensor for single-molecule αS assembly detection. RT-FAST allows the detection of the presence αS seeds WT and A53T variant in a given sample in only 90 min by adding a low quantity (35 µL at 100 nM) of recombinant αS for amplification. It also shows cross-seeding aggregation by adding mixing seeds A53T with WT monomers. Finally, we establish the dependence between the capture rate of aggregates by the nanopore sensor and the initial seed concentration from 200 pM to 2 pM, which promises further development toward a quantitative analysis of the initial seed concentration.

Bovine Respiratory Disease Considerations in Young Dairy Calves.

Raising young dairy calves presents many challenges for producers and veterinarians including losses attributable to BRD. This article will discuss several key concepts for practitioners to consider when applying evidence-based medicine for the control and treatment of BRD in young dairy calves. The authors review BRD complex, provide considerations for diagnostic approaches, and discuss research associated with the control and treatment of BRD.

Investigation of α-Synuclein and Amyloid-ß(42)-E22Δ Oligomers Using SiN Nanopore Functionalized with L-Dopa.

Solid-state nanopores are an emerging technology used as a high-throughput, label-free analytical method for the characterization of protein aggregation in an aqueous solution. In this work, we used Levodopamine to coat a silicon nitride nanopore surface that was fabricated through a dielectric breakdown in order to reduce the unspecific adsorption. The coating of inner nanopore wall by investigation of the translocation of heparin. The functionalized nanopore was used to investigate the aggregation of amyloid-ß and α-synuclein, two biomarkers of degenerative diseases. In the first application, we demonstrate that the α-synuclein WT is more prone to form dimers than the variant A53T. In the second one, we show for the Aß(42)-E22Δ (Osaka mutant) that the addition of Aß(42)-WT monomers increases the polymorphism of oligomers, while the incubation with Aß(42)-WT fibrils generates larger aggregates.

Conical nanopores highlight the pro-aggregating effects of pyrimethanil fungicide on Aß(1-42) peptides and dimeric splitting phenomena.

The Aß(1-42) aggregation is a key event in the physiopathology of Alzheimer's disease (AD). Exogenous factors such as environmental pollutants, and more particularly pesticides, can corrupt Aß(1-42) assembly and could influence the occurrence and pathophysiology of AD. However, pesticide involvement in the early stages of Aß(1-42) aggregation is still unknown. Here, we employed conical track-etched nanopore in order to analyse the Aß(1-42) fibril formation in the presence of pyrimethanil, a widely used fungicide belonging to the anilinopyrimidine class. Our results evidenced a pro-aggregating effect of pyrimethanil on Aß(1-42). Aß(1-42) assemblies were successfully detected using conical nanopore coated with PEG. Using an analytical model, the large current blockades observed (>0.7) were assigned to species with size close to the sensing pore. The long dwell times (hundreds ms scale) were interpreted by the possible interactions amyloid/PEG using molecular dynamic simulation. Such interaction could leave until splitting phenomena of the dimer structure. Our work also evidences that the pyrimethanil induce an aggregation of Aß(1-42) mechanism in two steps including the reorganization prior the elongation phase.

Solid-state and polymer nanopores for protein sensing: A review.

In two decades, the solid state and polymer nanopores became attractive method for the protein sensing with high specificity and sensitivity. They also allow the characterization of conformational changes, unfolding, assembly and aggregation as well the following of enzymatic reaction. This review aims to provide an overview of the protein sensing regarding the technique of detection: the resistive pulse and ionic diodes. For each strategy, we report the most significant achievement regarding the detection of peptides and protein as well as the conformational change, protein-protein assembly and aggregation process. We discuss the limitations and the recent strategies to improve the nanopore resolution and accuracy. A focus is done about concomitant problematic such as protein adsorption and nanopore lifetime.

Detection of Amyloid-ß Fibrils Using Track-Etched Nanopores: Effect of Geometry and Crowding.

Several neurodegenerative diseases have been linked to proteins or peptides that are prone to aggregate in different brain regions. Aggregation of amyloid-ß (Aß) peptides is recognized as the main cause of Alzheimer's disease (AD) progression, leading to the formation of toxic Aß oligomers and amyloid fibrils. The molecular mechanism of Aß aggregation is complex and still not fully understood. Nanopore technology provides a new way to obtain kinetic and morphological aspects of Aß aggregation at a single-molecule scale without labeling by detecting the electrochemical signal of the peptides when they pass through the hole. Here, we investigate the influence of nanoscale geometry (conical and bullet-like shape) of a track-etched nanopore pore and the effect of molecular crowding (polyethylene glycol-functionalized pores) on Aß fibril sensing and analysis. Various Aß fibril samples that differed by their length were produced by sonication of fibrils obtained in the presence of epigallocatechin gallate. The conical nanopore functionalized with polyethylene glycol (PEG) 5 kDa is suitable for discrimination of the fibril size from relative current blockade. The bullet-like-shaped nanopore enhances the amplitude of the current and increases the dwell time, allowing us to well discern the fibrils. Finally, the nanopore crowded with PEG 20 kDa enhances the relative current blockade and increases the dwell time; however, the discrimination is not improved compared to the "bullet-shaped" nanopore.

Machine Learning to Improve the Sensing of Biomolecules by Conical Track-Etched Nanopore.

Single nanopore is a powerful platform to detect, discriminate and identify biomacromolecules. Among the different devices, the conical nanopores obtained by the track-etched technique on a polymer film are stable and easy to functionalize. However, these advantages are hampered by their high aspect ratio that avoids the discrimination of similar samples. Using machine learning, we demonstrate an improved resolution so that it can identify short single- and double-stranded DNA (10- and 40-mers). We have characterized each current blockade event by the relative intensity, dwell time, surface area and both the right and left slope. We show an overlap of the relative current blockade amplitudes and dwell time distributions that prevents their identification. We define the different parameters that characterize the events as features and the type of DNA sample as the target. By applying support-vector machines to discriminate each sample, we show accuracy between 50% and 72% by using two features that distinctly classify the data points. Finally, we achieved an increased accuracy (up to 82%) when five features were implemented.

Cholecystohepatic Duct: A Biliary Duct Variant Resulting in Postcholecystectomy Bile Leak-Case Report and Review of Normal and Common Variant Biliary Anatomy.

Although relatively infrequent, bile duct leaks are among the primary complications of hepatobiliary surgery and cholecystectomy given the large number of these operations performed annually around the world. Variant biliary anatomy increases the risk of surgical complications, especially if unrecognized on preoperative imaging or intraoperatively. Presented here is a case of a patient with an unrecognized cholecystohepatic duct at the time of surgery leading to bile leak after cholecystectomy. Numerous factors made for a technically difficult surgery with obscuration of the true anatomy, ultimately resulting in transection of the cholecystohepatic duct. Understanding normal and variant biliary anatomy will help prevent avoidable complications of hepatobiliary surgery.

Diagnosis and Management of Rumen Acidosis and Bloat in Feedlots.

Ruminal acidosis and ruminal bloat represent the most common digestive disorders in feedlot cattle. Ruminants are uniquely adapted to digest and metabolize a large range of feedstuffs. Although cattle have the ability to handle various feedstuffs, disorders associated with altered ruminal fermentation can occur. Proper ruminal microorganism adaptation and a consistent substrate (ration) help prevent digestive disorders. Feed bunk management, sufficient ration fiber, consistent feed milling, and appropriate response to abnormal weather are additional factors important in prevention of digestive disorders. When digestive disorders are suspected, timely diagnosis is imperative.