The implementation of a hepatitis C testing service in community pharmacies: I-COPTIC consensus statement.

OBJECTIVES: This aimed to develop a blueprint for an effective community pharmacy Hepatitis C virus (HCV) testing service by producing a consensus statement. STUDY DESIGN: This was a modified Delphi process. METHODS: We recruited a heterogenous panel of experts (who had been involved in the setup or delivery of a community pharmacy HCV testing service) by purposive and chain referral methods. We had three rounds of a modified Delphi process. The first was a series of questions with free text responses and was analysed using thematic analysis, and the second and third were statements for the respondents to rate using a 7-point Likert scale. Consensus was predefined in a published protocol, and the results were reviewed by a public and patient involvement panel before the statement was finalised. RESULTS: We had 24 participants, including community and hospital-based pharmacists, local pharmaceutical committee members, charity representatives (Hepatitis C Trust), local clinical service lead, nurse specialists and doctors. The response rate of the first, second and third rounds were 100%, 96% and 88%, respectively. After the third round, we had 60 statements that reached consensus. We discussed the accepted statements with a patient and public involvement group. We used these statements to produce the I-COPTIC statement and a graphical summary. CONCLUSIONS: We developed a blueprint for the design of a gold standard community pharmacy HCV testing service. We believe this will support the successful implementation of community pharmacy testing for HCV. Community pharmacy testing is an important service to help achieve and maintain HCV elimination.

Non-invasive suppression of the human nucleus accumbens (NAc) with transcranial focused ultrasound (tFUS) modulates the reward network: a pilot study.

Background: The nucleus accumbens (NAc) is a key node of the brain reward circuit driving reward-related behavior. Dysregulation of NAc has been demonstrated to contribute to pathological markers of addiction in substance use disorder (SUD) making it a potential therapeutic target for brain stimulation. Transcranial focused ultrasound (tFUS) is an emerging non-invasive brain stimulation approach that can modulate deep brain regions with a high spatial resolution. However, there is currently no evidence showing how the brain activity of NAc and brain functional connectivity within the reward network neuromodulated by tFUS on the NAc. Methods: In this pilot study, we carried out a single-blind, sham-controlled clinical trial using functional magnetic resonance imaging (fMRI) to investigate the underlying mechanism of tFUS neuromodulating the reward network through NAc in ten healthy adults. Specifically, the experiment consists of a 20-min concurrent tFUS/fMRI scan and two 24-min resting-state fMRI before and after the tFUS session. Results: Firstly, our results demonstrated the feasibility and safety of 20-min tFUS on NAc. Additionally, our findings demonstrated that bilateral NAc was inhibited during tFUS on the left NAc compared to sham. Lastly, increased functional connectivity between the NAc and medial prefrontal cortex (mPFC) was observed after tFUS on the left NAc, but no changes for the sham group. Conclusion: Delivering tFUS to the NAc can modulate brain activations and functional connectivity within the reward network. These preliminary findings suggest that tFUS could be potentially a promising neuromodulation tool for the direct and non-invasive management of the NAc and shed new light on the treatment for SUD and other brain diseases that involve reward processing.

Comparative analysis of thermal indices for modeling cold and heat stress in US dairy systems.

Quantifying the impact of thermal stress on milk yields is essential to effectively manage present and future risks in dairy systems. Despite the existence of numerous heat indices designed to communicate stress thresholds, little information is available regarding the accuracy of different indices in estimating milk yield losses from both cold and heat stress at large spatio-temporal scales. To address this gap, we comparatively analyzed the performance of existing thermal indices in capturing US milk yield response to both cold and heat stress at the national scale. We selected four commonly used thermal indices: the Temperature and Humidity Index (THI), Black Globe Humidity Index (BGHI), Adjusted Temperature and Humidity Index (THIadj), and Comprehensive Climate Index (CCI). Using a statistical panel regression model with observational and reanalysis weather data from 1981-2020, we systematically compared the patterns of yield sensitivities and statistical performance of the four indices. We found that the US state-level milk yield variability was better explained by the THIadj and CCI, which combine the effects of temperature, humidity, wind, and solar radiation. Our analysis also reveals a continuous and nonlinear responses of milk yields to a range of cold to heat stress across all four indices. This implies that solely relying on fixed thresholds of these indices to model milk yield changes may be insufficient to capture cumulative thermal stress. Cold extremes reduced milk yields comparably to those impacted by heat extremes on the national scale. Additionally, we found large spatial variability in milk yield sensitivities, implying further limitations to the use of fixed thresholds across locations. Moreover, we found decreased yield sensitivity to thermal stress in the most recent two decades, suggesting adaptive changes in management to reduce weather-related risks.

Estimating Ejection Fraction from the 12 Lead ECG among Patients with Acute Heart Failure.

Background: Identifying patients with low left ventricular ejection fraction (LVEF) in the emergency department using an electrocardiogram (ECG) may optimize acute heart failure (AHF) management. We aimed to assess the efficacy of 527 automated 12-lead ECG features for estimating LVEF among patients with AHF. Method: Medical records of patients >18 years old and AHF-related ICD codes, demographics, LVEF %, comorbidities, and medication were analyzed. Least Absolute Shrinkage and Selection Operator (LASSO) identified important ECG features and evaluated performance. Results: Among 851 patients, the mean age was 74 years (IQR:11), male 56% (n=478), and the median body mass index was 29 kg/m2 (IQR:1.8). A total of 914 echocardiograms and ECGs were matched; the time between ECG-Echocardiogram was 9 hours (IQR of 9 hours); ≤30% LVEF (16.45%, n=140). Lasso demonstrated 42 ECG features important for estimating LVEF ≤30%. The predictive model of LVEF ≤30% demonstrated an area under the curve (AUC) of 0.86, a 95% confidence interval (CI) of 0.83 to 0.89, a specificity of 54% (50% to 57%), and a sensitivity of 91 (95% CI: 88% to 96%), accuracy 60% (95% CI:60 % to 63%) and, negative predictive value of 95%. Conclusions: An explainable machine learning model with physiologically feasible predictors may be useful in screening patients with low LVEF in AHF.

Disparities in 30-Day Readmission Between Medicare/Medicaid and Private Insurance Among Patients With Heart Failure Screened for Cognitive Impairment.

BACKGROUND: Racial disparities exist among patients with heart failure (HF). HF is often comorbid with cognitive impairment. Appropriate self-care can prevent HF hospital readmissions but requires access to resources through insurance. Racial differences exist between insurance types, and this may influence the disparity between races and patients with HF and cognitive impairment. OBJECTIVE: The objectives of this study were to examine the relationships between insurance type and self-care stratified by race and to assess for differences in time-to-30-day readmission among patients with HF with cognitive impairment. METHODS: This is a secondary analysis of data collected among hospitalized patients with HF with cognitive impairment. Patients completed surveys on self-care (Self-Care of Heart Failure Index), HF knowledge (Dutch Heart Failure Knowledge Scale), depression (Geriatric Depression Scale), and social support (Enhancing Recovery in Coronary Heart Disease Social Support Inventory). Socioeconomic data were collected. Linear models were created to examine the relationships between insurance type and self-care by race. Kaplan-Meier curves and Cox regression were used to assess readmission. RESULTS: The sample of 125 patients with HF with cognitive impairment was predominantly Black (68%, n = 85) and male (53%, n = 66). The sample had either Medicare/Medicaid (62%, n = 78) or private insurance (38%, n = 47). Black patients with HF with cognitive impairment and private insurance reported higher self-care confidence compared with Black patients with HF with cognitive impairment and Medicare/Medicaid (P < .05). Medicare/Medicaid was associated with a higher frequency of 30-day readmission and a faster time-to-readmission. CONCLUSIONS: Patients with HF with cognitive impairment and Medicare/Medicaid insurance reported lower self-care confidence and more likely to be readmitted within 30 days.

Development of a high-throughput scale-down model in Ambr® 250 HT for plasmid DNA fermentation processes.

Recent advances in messenger ribonucleic acid (mRNA) vaccines and gene therapy vectors have increased the need for rapid plasmid DNA (pDNA) screening and production within the biopharmaceutical industry. High-throughput (HT) fermentor systems, such as the Ambr® 250 HT, can significantly accelerate process development timelines of pDNA upstream processes compared to traditional bench-scale glass fermentors or small-scale steam-in-place (SIP) fermentors. However, such scale-down models must be qualified to ensure that they are representative of the larger scale process similar to traditional small-scale models. In the current study, we developed a representative scale-down model of a Biostat® D-DCU 30 L pDNA fermentation process in Ambr® 250 HT fermentors using three cell lines producing three different constructs. The Ambr scale-down model provided comparable process performance and pDNA quality as the 30 L SIP fermentation process. In addition, we demonstrated the predictive value of the Ambr model by two-way qualification, first by accurately reproducing the prior trends observed in a 30 L process, followed by predicting new process trends that were then successfully reproduced in the 30 L process. The representative and predictive scale-down Ambr model developed in this study would enable a faster and more efficient approach to strain/clone/host-cell screening, pDNA process development and characterization studies, process scale-up studies, and manufacturing support.

Higher Doses of Calcium Associated With Survival in Trauma Patients.

INTRODUCTION: Calcium is required for coagulation, cardiac output, and peripheral vascular resistance. Between 85% and 94% of trauma patients treated with massive blood transfusion develop hypocalcemia.1 The aim of this study is to evaluate the relationship between increased intravenous calcium administration during massive transfusion and improved survival of trauma patients. METHODS: We performed a retrospective analysis of trauma patients who received massive transfusion over a 2-y period. Doses of elemental calcium administered per unit of blood product transfused were calculated by calcium to blood product ratio (CBR). Chi-square test evaluated association between coagulopathy and 30-d mortality. Two-sample t-test evaluated association between CBR and coagulopathy. Bivariate regression analysis evaluated association between CBR and blood products transfused per patient. Multivariable logistic regression analysis, controlling for age, sex, coagulopathy, and Injury Severity Score evaluated the association between CBR and mortality. RESULTS: The study included 77 patients. Coagulopathy was associated with increased 30-d mortality (P < 0.05). Patients who survived had higher CBR than those who died (P < 0.05). CBR was associated with a significant reduction in total blood products transfused per patient (P < 0.05). CBR was not associated with coagulopathy (P = 0.24). Multivariable logistic regression analysis demonstrated that Injury Severity Score ≥16, coagulopathy and decreased CBR were significant predictors of mortality (P < 0.05). CBR above 50 mg was a predictor of survival (P < 0.05). CONCLUSIONS: Higher doses of calcium given per blood product transfused were associated with improved 30-d survival and decreased blood product transfusions.

Trends in NIH-funding to schools of nursing in the last 17 years.

BACKGROUND: Growing clinical demands, faculty retirements, fewer PhD-prepared graduates, and funding instability are challenges for nursing science. PURPOSE: The purpose of this analysis was to investigate National Institutes of Health (NIH) funding patterns in schools of nursing (SONs). METHODS: Data were extracted from the Blue Ridge Institute for Medical Research between 2006 and 2022. Growth modeling examined changes in funding over time between private and public SONs. DISCUSSION: In the last 17 years, NIH funding for SONs has risen nearly 25% but remains only 1% of the total NIH budget for extramural research. Overall, 109 (75%) of the SONs were public and 36 (25%) were private institutions. Regarding geography, 90% of the States received NIH funding except six: ID, ME, MS, NH, VT, and WY. Private SONs consistently received more funding than public SONs but the difference was only statistically significant in 2022. CONCLUSION: NIH funding has significantly increased to SONs, there is better geographic distribution but a funding disparity exists between public and private SONs.

Misfolded protein oligomers: mechanisms of formation, cytotoxic effects, and pharmacological approaches against protein misfolding diseases.

The conversion of native peptides and proteins into amyloid aggregates is a hallmark of over 50 human disorders, including Alzheimer's and Parkinson's diseases. Increasing evidence implicates misfolded protein oligomers produced during the amyloid formation process as the primary cytotoxic agents in many of these devastating conditions. In this review, we analyze the processes by which oligomers are formed, their structures, physicochemical properties, population dynamics, and the mechanisms of their cytotoxicity. We then focus on drug discovery strategies that target the formation of oligomers and their ability to disrupt cell physiology and trigger degenerative processes.

The Effect of Mandatory Fitness Requirements on Cardiovascular Events: A State-by-State Analysis Using a National Database.

BACKGROUND: Cardiovascular events are known to be the leading cause of death among on-duty firefighters. Implementing fitness standards may help reduce the incidence of cardiovascular deaths; however, standards vary between firefighter type and states. We aimed to investigate the rate of cardiovascular events among firefighters across states. METHODS: Using publicly available data from the United States Fire Administration, we explored the rates of cardiovascular deaths between firefighter type (e.g., career, volunteer, and wildland) and state. Specifically, we examined rates of cardiovascular deaths between California and Tennessee, which have fitness standards for all firefighters, and New York, which does not have fitness standards for volunteer firefighters. We used descriptive statistics and trend analysis to examine the data. FINDINGS: Most cardiovascular events occur among volunteer firefighters (60.6%, n = 877). Volunteer firefighters had 7.5 (95% CI = [4.8, 11.7], p < .001) greater odds of cardiovascular events compared to wildland firefighters, who had the lowest incidence of cardiovascular events (1.7%, n = 24). New York reported the most cardiovascular events (n = 161), primarily among volunteer firefighters (73.9%, n = 119). After the passage of legislation mandating fitness standards in California, a downtrend in the number of volunteer firefighter fatalities is observed. However, a null effect was observed in Tennessee after the passage of similar fitness standards as in California. CONCLUSIONS/APPLICATIONS TO PRACTICE: Volunteer firefighters are significantly more likely to die of a cardiovascular event than career and wildland firefighters, both of which have stricter fitness standards. However, the effect of legislation mandating stricter fitness standards among volunteers did not produce a clear benefit for preventing fatalities. Nurses need to promote cardiovascular health among volunteer firefighters.

A Scoping Review of Varying Mobile Electrocardiographic Devices.

The electrocardiogram (ECG) can now be measured using mobile devices. Mobile ECG devices, which are defined as devices capable of recording and transmitting non-standard ECGs, offer numerous advantages such as cost-effectiveness and being user-friendly. Mobile ECG can also extend recording lengths (e.g., 2 days, 14 days), which is necessary to capture important intermittent events (e.g., cardiac arrhythmias) and evaluate prognostic risk markers (e.g., prolonged corrected QT (QTc) interval). Some mobile ECG devices can even connect to broadband networks allowing patients to remotely transmit their ECG to a clinician. This article systematically examines different mobile ECG devices used in prior studies and provides a detailed assessment of five diverse yet commonly used mobile ECG devices: AliveCor KardiaMobile; AliveCor KardiaMobile 6L; iRhythm ZioPatch; Apple Smartwatch ECG; and CardioSecur System. These mobile ECG devices are diverse in the number of leads measured and the duration of monitoring. Similar to their diversity, there has been a wide range of clinical applications of mobile ECG devices. Despite significant progress, questions regarding data quality, and clinican and patient acceptance and compliance persist.

Impact of Renal Access Angle and Speed of Nephroscope Retrieval Movements on the Vortex Effect.

OBJECTIVE: To analyze the influence of different renal access angles (AAs) and nephroscope retrieval speeds on the efficacy of the vortex effect (VE) in mini-percutaneous nephrolithotomy (mini-PCNL). This study aimed to understand the poorly understood physical components of the VE. MATERIALS AND METHODS: A Pexiglas™ (KUS®) model was built based on the dimensions of a 15/16 F mini-PCNL set (Karl Storz). The flow rate was continuous via an automatic pump and calibrated to achieve hydrodynamic equivalence to the real equipment. One experiment consisted of manually retrieving all 30 stone phantoms (3 mm diameter) utilizing only the VE. Cumulative time to retrieve all stones was measured. An accelerometer recorded instant speeds of the nephroscope every 0.08 seconds (s), and 3 experiments were performed at each angle (0°, 45°, and 90°). A logistic regression model was built utilizing maximum speeds and access angles to predict the effectiveness of the VE. RESULTS: Mean cumulative time for complete stone retrieval was 28.1 seconds at 0° vs 116.5 seconds at 45° vs 101.4 seconds at 90° (P < .01). We noted significantly higher speeds at 0° compared to 45° and 90° (P < .01); however, differences in average and maximum speed between 45° and 90° were not statistically significant (P = .21 and P = .25, respectively). The regression model demonstrated a negative association between increasing maximum speed and VE's effectiveness (OR 0.547, CI 95% 0.350-0.855, P < .01). When controlling for maximum speed, the 0° angle had significantly higher chances of achieving at least a partially effective VE. CONCLUSION: Increasing the renal access angle or nephroscope extraction speed negatively impacts the effectiveness of the VE. This significantly increased procedure time in the laboratory model, suggesting that the VE is less effective at higher sheath angles.

Signs and Symptoms Clusters Among Patients With Acute Heart Failure: A Correlational Study.

BACKGROUND: Patients with acute heart failure present to the emergency department with a myriad of signs and symptoms. Symptoms evaluated in clusters may be more meaningful than those evaluated individually by clinicians. OBJECTIVE: Among emergency department patients, we aimed to identify signs and symptoms correlations, clusters, and differences in clinical variables between clusters. METHODS: Medical record data included adults older than 18 years, International Classification of Diseases, Tenth Revisions codes , and positive Framingham Heart Failure Diagnostic Criteria. Exclusion criteria included medical records with a ventricular assist device and dialysis. For analysis, correlation, and the Gower distance, the independent t test, Mann-Whitney U test, χ 2 test, and regression were performed. RESULTS: A secondary analysis was conducted from the data set to evaluate door-to-diuretic time among patients with acute heart failure in the emergency department. A total of 218 patients were included, with an average age of 69 ± 15 years and predominantly White (74%, n = 161). Two distinct symptom clusters were identified: severe and mild congestion. The severe congestion cluster had a more comorbidity burden compared with the mild congestion cluster, as measured by the Charlson Comorbidity index (cluster 1 vs cluster 2, 6 [5-7] vs 5 [4-6]; P = .0019). Heart failure with preserved ejection fraction was associated with the severe congestion symptom cluster ( P = .009), and heart failure with mildly reduced ejection fraction was associated with the mild congestion cluster ( P = .019). CONCLUSIONS: In conclusion, 2 distinct symptom clusters were identified among patients with acute heart failure. Symptom clusters may be related to ejection fraction or overall cardiac output and comorbidity burden.

TubULAR: tracking in toto deformations of dynamic tissues via constrained maps.

A common motif in biology is the arrangement of cells into tubes, which further transform into complex shapes. Traditionally, analysis of dynamic tissues has relied on inspecting static snapshots, live imaging of cross-sections or tracking isolated cells in three dimensions. However, capturing the interplay between in-plane and out-of-plane behaviors requires following the full surface as it deforms and integrating cell-scale motions into collective, tissue-scale deformations. Here, we present an analysis framework that builds in toto maps of tissue deformations by following tissue parcels in a static material frame of reference. Our approach then relates in-plane and out-of-plane behaviors and decomposes complex deformation maps into elementary contributions. The tube-like surface Lagrangian analysis resource (TubULAR) provides an open-source implementation accessible either as a standalone toolkit or as an extension of the ImSAnE package used in the developmental biology community. We demonstrate our approach by analyzing shape change in the embryonic Drosophila midgut and beating zebrafish heart. The method naturally generalizes to in vitro and synthetic systems and provides ready access to the mechanical mechanisms relating genetic patterning to organ shape change.

A comparison of energy expenditure and perceived exertion between standard axillary crutches, knee scooters, and a hands-free crutch.

BACKGROUND: Ambulation using standard axillary crutches (SACs) is associated with increased energy expenditure (EE) and decreased ability to perform activities of daily living (ADLs). Using a hands-free crutch (HFC) displays potential for easier completion of ADLs and reduction in energy requirements. OBJECTIVES: To determine if a HFC elicits lower EE and heart rate (HR), improvement in performance of ADLs, and decreased rating of perceived exertion (RPE) compared to common ambulatory devices. DESIGN: A randomized crossover-controlled trial. SETTING: University community. PARTICIPANTS: Twenty healthy college students. MAIN OUTCOME MEASURES: Participants completed a 6-minute walk test at 50 m/min, an ADLs course, and a two-flight stair climb with SACs, HFC, knee scooter (KS), and unassisted ambulation (UA). The order of trial conditions was randomized. EE, HR, time to complete ADLs course and stair climb, and RPE during each condition were obtained. One-way analyses of variance were performed to compare EE, HR response, and RPE between the assistive devices and UA. RESULTS: In all outcomes UA resulted in lower EE, HR, and RPE compared to all the assistive devices (p < .05). For the ADLs course, EE was the same for the three assistive devices, whereas HR was significantly lower for HFC compared to SACs and KS (p < .05). RPE for HFC and KS was lower than SACs (p < .05). For the 6MWT, each device significantly differed from the other devices for EE, HR, and RPE, with KS eliciting the lowest values, followed by HFC. For the stair climbing task, HFC elicited lower EE, HR, and RPE than SACs. Fourteen participants indicated their overall preference for HFCs. CONCLUSIONS: In individuals prescribed weight-bearing restrictions, using a HFC may offer an easier and more preferred alternative to more commonly used SACs during ambulation, stair climbing, and other ADLs.

Active cell divisions generate fourfold orientationally ordered phase in living tissue.

Morphogenesis, the process through which genes generate form, establishes tissue-scale order as a template for constructing the complex shapes of the body plan. The extensive growth required to build these ordered substrates is fuelled by cell proliferation, which, naively, should destroy order. Understanding how active morphogenetic mechanisms couple cellular and mechanical processes to generate order-rather than annihilate it-remains an outstanding question in animal development. We show that cell divisions are the primary drivers of tissue flow, leading to a fourfold orientationally ordered phase. Waves of anisotropic cell proliferation propagate across the embryo with precise patterning. Defects introduced into the nascent lattice by cell divisions are moved out of the tissue bulk towards the boundary by subsequent divisions. Specific cell proliferation rates and orientations enable cell divisions to organize rather than fluidize the tissue. We observe this using live imaging and tissue cartography to analyse the dynamics of fourfold tissue ordering in the trunk segmental ectoderm of the crustacean Parhyale hawaiensis beginning 72 h after egg lay. The result is a robust, active mechanism for generating global orientational order in a non-equilibrium system that sets the stage for the subsequent development of shape and form.

Bed nucleus of the stria terminalis GABA neurons are necessary for changes in foraging behaviour following an innate threat.

Foraging is a universal behaviour that has co-evolved with predation pressure. We investigated the role of the bed nucleus of the stria terminalis (BNST) GABA neurons in robotic and live predator threat processing and their consequences in post-threat encounter foraging. Both robotic and live predator interactions increased BNST GABA neuron activity. Mice were trained to procure food in a laboratory-based foraging apparatus in which food pellets were placed at incrementally greater distances from a nest zone. After mice learned to forage, they were exposed to a robotic or live predator threat, while BNST GABA neurons were chemogenetically inhibited. Post-robotic threat encounter, mice spent more time in the nest zone, but other foraging parameters were unchanged compared with pre-encounter behaviour. Inhibition of BNST GABA neurons had no effect on foraging behaviour post-robotic threat encounter. Following live predator exposure, control mice spent significantly more time in the nest zone, increased their latency to successfully forage, and significantly altered their overall foraging performance. Inhibition of BNST GABA neurons during live predator exposure prevented changes in foraging behaviour from developing after a live predator threat. BNST GABA neuron inhibition did not alter foraging behaviour during robotic or live predator threats. We conclude that these results demonstrate that while both robotic and live predator encounters effectively intrude on foraging behaviour, the perceived risk and behavioural consequences of the threat are distinguishable. Additionally, BNST GABA neurons may play a role in the integration of prior innate predator threat experience that results in hypervigilance during post-encounter foraging behaviour.

Smell and Taste Disorders in Primary Care.

Disorders of smell and taste are reported by approximately one-fifth of people 40 years and older, and one-third of people 80 years and older. These disorders affect quality of life and the ability to identify smoke and toxins. Smell and taste disorders can be early signs of dementia or Parkinson disease and are associated with increased mortality. Dysfunction may be apparent or may develop insidiously. Screening questionnaires are available, but many patients are unaware of their disorder. Most smell and taste disorders are due to sinonasal disease but also could be caused by smoking, medications, head trauma, neurodegenerative disease, alcohol dependence, or less common conditions. The differential diagnosis should guide the evaluation and include anterior rhinoscopy and an examination of the oral cavity, head, and cranial nerves. Further investigation is often unnecessary, but nasal endoscopy and computed tomography of the sinuses may be helpful. Magnetic resonance imaging of the head with contrast should be performed if there is an abnormal neurologic examination finding or if trauma or a tumor is suspected. Olfactory testing is indicated in refractory cases or for patients with poor quality of life and disease associated with smell or taste dysfunction. Smell and taste disorders may resolve when reversible causes are treated, but improvement is less likely when they are due to trauma, age, or neurodegenerative disease. Olfactory training is a self-administered mindful exposure therapy that may improve olfactory function. Physicians should encourage patients to ensure that smoke and other alarms are operational and to adhere to food expiration dates.

Secondary Processes Dominate the Quiescent, Spontaneous Aggregation of α-Synuclein at Physiological pH with Sodium Salts.

The accurate recapitulation in an in vitro assay of the aggregation process of α-synuclein in Parkinson's disease has been a significant challenge. As α-synuclein does not aggregate spontaneously in most currently used in vitro assays, primary nucleation is triggered by the presence of surfaces such as lipid membranes or interfaces created by shaking, to achieve aggregation on accessible time scales. In addition, secondary nucleation is typically only observed by lowering the pH below 5.8. Here we investigated assay conditions that enables spontaneous primary nucleation and secondary nucleation at pH 7.4. Using 400 mM sodium phosphate, we observed quiescent spontaneous aggregation of α-synuclein and established that this aggregation is dominated by secondary processes. Furthermore, the presence of potassium ions enhanced the reproducibility of quiescent α-synuclein aggregation. This work provides a framework for the study of spontaneous α-synuclein aggregation at physiological pH.