Digital assay for rapid electronic quantification of clinical pathogens using DNA nanoballs.

Fast and accurate detection of nucleic acids is key for pathogen identification. Methods for DNA detection generally rely on fluorescent or colorimetric readout. The development of label-free assays decreases costs and test complexity. We present a novel method combining a one-pot isothermal generation of DNA nanoballs with their detection by electrical impedance. We modified loop-mediated isothermal amplification by using compaction oligonucleotides that self-assemble the amplified target into nanoballs. Next, we use capillary-driven flow to passively pass these nanoballs through a microfluidic impedance cytometer, thus enabling a fully compact system with no moving parts. The movement of individual nanoballs is detected by a change in impedance providing a quantized readout. This approach is flexible for the detection of DNA/RNA of numerous targets (severe acute respiratory syndrome coronavirus 2, HIV, ß-lactamase gene, etc.), and we anticipate that its integration into a standalone device would provide an inexpensive (<$5), sensitive (10 target copies), and rapid test (<1 hour).

Emergency Department Point-of-care Ultrasound Identification of Suspected Lemierre's Syndrome: A Case Report.

INTRODUCTION: Lemierre's syndrome is septic thrombophlebitis of the internal jugular vein, most commonly associated with head and neck infections. While central catheters are associated with venous thromboembolism and catheter-associated bacterial infections, cases of Lemierre's syndrome caused by central catheters are extraordinarily rare. CASE REPORT: We detail a case of Lemierre's syndrome resulting from a peripherally inserted central catheter in a pregnant female patient. Diagnosis of this rare and potentially life-threatening disease process was expedited using point-of-care ultrasound. CONCLUSION: Diagnosis of rare but potentially life- or limb-threatening pathologies is paramount to the successful practice of emergency medicine. Identifying these rare disease processes requires a high index of suspicion and a work-up strategy that includes consideration of medical history in combination with lab and imaging findings to determine appropriate interventions.

The Use of Whole Blood in Rural Trauma Leads to Decreased Resource Utilization.

BACKGROUND: Nearly 40% of trauma deaths result from uncontrolled hemorrhage. Most of these deaths occur within 24 hours, highlighting the importance of early resuscitation. Balanced component resuscitation has been shown to improve outcomes in hemorrhagic shock. However, hemostatic properties may then be decreased, leading to inadequate coagulopathy treatment or higher transfusion requirements. Data comparing the efficacy of component vs. whole blood (WB) resuscitation in early trauma is poor, particularly in the rural population. This study investigates WB use and resource utilization at a rural Level 1 trauma center. METHODS: A prospective cohort study with historical controls (HC) was performed using patients over age 17 presenting as the highest priority trauma. Two units of WB were available to patients with signs of hemorrhagic shock, with subsequent transfusions via massive transfusion protocol or thromboelastography guidance. Component utilization, time to hemorrhage control, complications, and transfer times were examined. RESULTS: Forty patients received WB vs. 153 HC. WB patients had lower complication rates (35% vs. 55.6%; P = .02), and a significant reduction in pRBC utilization in the emergency department (0 vs. 2; P < .0001) and throughout admission (2.0 vs. 4.0; P = .0003). All patients had prolonged transport times given the rural setting (1.42 hours HC vs. 2.03 hours WB; P = .002). DISCUSSION: Unlike most urban WB studies, this study occurred in a rural area with extended transportation times, when WB is inaccessible for patients. Despite this delay, WB patients demonstrated lower component utilization and complication rates. Further research is needed to characterize the impact of early WB access.

Development of a multicentre automated model to reduce planning variability in radiotherapy of prostate cancer.

BACKGROUND AND PURPOSE: Inter-institutional studies highlighted correlation between consistent radiotherapy quality and improved overall patient survival. In treatment planning automation has the potential to address differences due to user-experience and training, promoting standardisation. The aim of this study was to evaluate implementation and clinical effect of a multicentre collaboratively-developed automated planning model for Intensity-Modulated Radiation Therapy/Volumetric-Modulated Arc Therapy of prostate. The model was built using a variety of public institutions' clinical plans, incorporating different contouring and dose protocols, aiming at minimising their variation. METHODS AND MATERIALS: A model using 110 clinically approved and treated prostate plans provided by different radiotherapy centres was built with RapidPlan (RP), for use on intact and post-prostatectomy prostate cases. The model was validated, distributed and introduced into clinical practice in all institutions. To investigate its impact a total of 126 patients, originally manually inverse planned (OP), were replanned using RP without additional planner manual intervention. Target and organ-at-risk (OAR) metrics were statistically compared between original and automated plans. RESULTS: For all centres combined and individually, RP provided plans comparable or superior to OP for all dose metrics. Statistically significant reductions with RP were found in bladder (V40Gy) and rectal (V50Gy) low doses (within 2.3% and 3.4% for combined and 4% and 10% individually). No clinically significant changes were seen for the PTV, independently of seminal vesicle inclusion. CONCLUSION: This project showed it is feasible to develop, share and implement RP models created with plans from different institutions treated with a variety of techniques and dose protocols, with the potential of improving treatment planning results and/or efficiency despite the original variability.

Porous AlGaN-Based Ultraviolet Distributed Bragg Reflectors.

Utilising dislocation-related vertical etching channels in gallium nitride, we have previously demonstrated a simple electrochemical etching (ECE) process that can create layered porous GaN structures to form distributed Bragg reflectors for visible light at wafer scale. Here, we apply the same ECE process to realise AlGaN-based ultraviolet distributed Bragg reflectors (DBRs). These are of interest because they could provide a pathway to non-absorbing UV reflectors to enhance the performance of UV LEDs, which currently have extremely low efficiency. We have demonstrated porous AlGaN-based UV DBRs with a peak reflectance of 89% at 324 nm. The uniformity of these devices is currently low, as the as-grown material has a high density of V-pits and these alter the etching process. However, our results indicate that if the material growth is optimised, the ECE process will be useful for the fabrication of UV reflectors.

DNA assembly with error correction on a droplet digital microfluidics platform.

BACKGROUND: Custom synthesized DNA is in high demand for synthetic biology applications. However, current technologies to produce these sequences using assembly from DNA oligonucleotides are costly and labor-intensive. The automation and reduced sample volumes afforded by microfluidic technologies could significantly decrease materials and labor costs associated with DNA synthesis. The purpose of this study was to develop a gene assembly protocol utilizing a digital microfluidic device. Toward this goal, we adapted bench-scale oligonucleotide assembly methods followed by enzymatic error correction to the Mondrian™ digital microfluidic platform. RESULTS: We optimized Gibson assembly, polymerase chain reaction (PCR), and enzymatic error correction reactions in a single protocol to assemble 12 oligonucleotides into a 339-bp double- stranded DNA sequence encoding part of the human influenza virus hemagglutinin (HA) gene. The reactions were scaled down to 0.6-1.2 µL. Initial microfluidic assembly methods were successful and had an error frequency of approximately 4 errors/kb with errors originating from the original oligonucleotide synthesis. Relative to conventional benchtop procedures, PCR optimization required additional amounts of MgCl2, Phusion polymerase, and PEG 8000 to achieve amplification of the assembly and error correction products. After one round of error correction, error frequency was reduced to an average of 1.8 errors kb- 1. CONCLUSION: We demonstrated that DNA assembly from oligonucleotides and error correction could be completely automated on a digital microfluidic (DMF) platform. The results demonstrate that enzymatic reactions in droplets show a strong dependence on surface interactions, and successful on-chip implementation required supplementation with surfactants, molecular crowding agents, and an excess of enzyme. Enzymatic error correction of assembled fragments improved sequence fidelity by 2-fold, which was a significant improvement but somewhat lower than expected compared to bench-top assays, suggesting an additional capacity for optimization.

A comparison of physical activity, physical fitness levels, BMI and blood pressure of adults with intellectual disability, who do and do not take part in Special Olympics Ireland programmes: Results from the SOPHIE study.

People with an intellectual disability are less physically active, live more sedentary lives, have lower fitness levels and are more likely to be overweight or obese than the general population. No evidence exists on the impact of participation in Special Olympics Ireland (SOI) on physical activity and physical fitness levels. Adults with intellectual disabilities (16-64 years) were recruited from services and SOI clubs. Physical measures included waist circumference, height, weight, blood pressure, heart rate and 6-min walking test. Self-report questionnaires gathered data on physical activity levels. Actigraph (GT3X) accelerometers were used to gain an objective measure of physical activity. SOI participants accumulated more moderate to vigorous physical activity per day, had higher fitness levels and more positive health profile scores than those not taking part in SOI. SOI has the potential to make a positive difference to people's physical health and subsequently their overall health and well-being.

Scalable Device for Automated Microbial Electroporation in a Digital Microfluidic Platform.

Electrowetting-on-dielectric (EWD) digital microfluidic laboratory-on-a-chip platforms demonstrate excellent performance in automating labor-intensive protocols. When coupled with an on-chip electroporation capability, these systems hold promise for streamlining cumbersome processes such as multiplex automated genome engineering (MAGE). We integrated a single Ti:Au electroporation electrode into an otherwise standard parallel-plate EWD geometry to enable high-efficiency transformation of Escherichia coli with reporter plasmid DNA in a 200 nL droplet. Test devices exhibited robust operation with more than 10 transformation experiments performed per device without cross-contamination or failure. Despite intrinsic electric-field nonuniformity present in the EP/EWD device, the peak on-chip transformation efficiency was measured to be 8.6 ± 1.0 × 108 cfu·µg-1 for an average applied electric field strength of 2.25 ± 0.50 kV·mm-1. Cell survival and transformation fractions at this electroporation pulse strength were found to be 1.5 ± 0.3 and 2.3 ± 0.1%, respectively. Our work expands the EWD toolkit to include on-chip microbial electroporation and opens the possibility of scaling advanced genome engineering methods, like MAGE, into the submicroliter regime.

An examination of the nutritional intake and anthropometric status of individuals with intellectual disabilities: Results from the SOPHIE study.

The prevalence of obesity appears greater in people with intellectual disabilities than those in the general population. This study aimed to examine the nutritional intake and anthropometric status of individuals with intellectual disabilities. Participants aged 16-64 years were recruited from intellectual disability service provider organizations ( n = 131). Data were collected using questionnaires; 4-day food dairies and weight, height and waist circumference measurements. Participants' mean body mass index (BMI) was 29.4 kg/m2 ± 6.1, 2.4% were underweight, 22.6% were normal weight, 28.2% were overweight and 46.8% were obese. Having a diagnosis of Down syndrome ( p = 0.03) was associated with increasing BMI. Increasing waist circumference was associated with increasing severity of ID ( p = 0.04). The mean-reported energy intake was 1890 kcal/day. Mean energy intakes from sugar, fat and saturated fat were above recommendations and few participants met micronutrient recommended daily amounts. This study highlights the alarming prevalence of overweight and obesity and poor diet quality of individuals with intellectual disabilities.

People with intellectual disability and their families' perspectives of Special Olympics Ireland: Qualitative findings from the SOPHIE study.

Previous research has highlighted that while involvement in Special Olympics can have benefits for athletes and their families, there can also be many barriers to participation. This qualitative study, which was part of a large mixed-method study, examined the experiences and perspectives of people with intellectual disability, their families and staff who work with them, about Special Olympics Ireland (SOI). A total of 47 participants (15 athletes, 6 non-athletes, 18 family members and 8 staff members) participated in focus group and individual interviews. Supplemental data, gathered as part of the larger study extracted from open-ended survey questions completed by 97 family members also informed this element. Findings revealed four main themes: impact of participation on athletes, impact of involvement on families, barriers to participation and how to enhance participation rates. Involvement in Special Olympics impacted positively on the quality of life of athletes and families. Enhanced availability of user-friendly information and service accessibility were important drivers identified for enhancing participation rates in Special Olympics.

Five-year efficacy and safety data of exenatide once weekly: long-term results from the DURATION-1 randomized clinical trial.

OBJECTIVE: To evaluate the 5-year efficacy and safety of once weekly exenatide. PATIENTS AND METHODS: The Diabetes Therapy Utilization: Researching Changes in A1C, Weight and Other Factors Through Intervention with Exenatide Once Weekly (DURATION-1) randomized clinical trial consisted of a 30-week controlled phase (2 mg of exenatide once weekly vs 10 µg of exenatide twice daily) with an open-ended uncontrolled extension (once weekly exenatide only) in patients with type 2 diabetes mellitus on background glucose-lowering therapies (April 15, 2006, through February 21, 2012). At week 30, patients initially receiving 10 µg of exenatide twice daily switched to 2 mg of exenatide once weekly. Study end points included changes from baseline in hemoglobin A1c, fasting plasma glucose, weight, lipids, and blood pressure. Long-term safety data included adverse events, liver and renal function, and heart rate. RESULTS: Of 258 extension-phase patients, 153 (59.3%) completed 5 years of treatment. Hemoglobin A1c levels were significantly and durably reduced from baseline (least-squares mean, -1.6%; 95% CI, -1.8% to -1.4%; vs -1.9% for exenatide once weekly at week 30), and 65 (43.9%) of 148 patients achieved hemoglobin A1c levels of less than 7.0%. Significant improvements in fasting plasma glucose level (-28.8 mg/dL; 95% CI, -36.2 to -21.5 mg/dL), weight (-3.0 kg; 95% CI, -4.6 to -1.3 kg), lipids, and diastolic blood pressure were observed, with minimal heart rate increase. Frequencies of nausea and injection-site reactions or nodules were decreased vs the initial 30-week controlled phase. Minor hypoglycemia occurred predominantly with sulfonylurea use, and no major hypoglycemia or new safety signals were observed. CONCLUSION: Long-term once weekly exenatide treatment was generally well tolerated with sustained glycemic improvement, weight reduction, and improved markers of cardiovascular risk in patients with type 2 diabetes. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00308139.

Immediate and 1-year chest compression quality: effect of instantaneous feedback in simulated cardiac arrest.

INTRODUCTION: Several studies have demonstrated subpar chest compression (CC) performance by trained health care professionals. The objective of this study was to determine the immediate and sustained effect of instantaneous audiovisual feedback on CC quality. METHODS: A prospective, randomized, crossover study measuring the effect of audiovisual feedback training on the performance of CCs by health care providers and medical students in a simulated cardiopulmonary arrest scenario was performed. Compression rate, hand placement, depth, and recoil were collected using 60-second epochs of CC on a simulation mannequin. RESULTS: Data from 200 initial enrollments and 100 tested 1 year later were analyzed by evaluators using standard criterion. At initial testing, feedback trainees demonstrated significantly improved depth compliance, recoil compliance, and accuracy of hand placement. One year later, the previous year's control group now receiving feedback demonstrated immediate improvement in depth, hand placement, and rate. In the feedback group, the only statistically significant improvement from initial baseline to the baseline 1 year later was an 18% improvement in depth compliance. However, the same improvement rate was seen in the control group. Improved depth compliance performance was correlated to the number of cardiopulmonary resuscitation training sessions received external to the study. CONCLUSIONS: Instantaneous audiovisual feedback training on CC quality produces immediate improvements in compression rate, hand placement, as well as depth and recoil compliance. These improvements, however, are not retained 1 year later. Improved depth performance may be correlated to an increased training frequency.

Caffeine overdose resulting in severe rhabdomyolysis and acute renal failure.

Despite the fact that caffeine is the most commonly used stimulant in modern society, cases of caffeine overdose are relatively rare, with fatalities reported from doses of 10 g and higher (Nord J Psychiatry. 2006;60:97-106). Large doses produce symptoms associated with stimulation of the cardiovascular, central nervous, and gastrointestinal symptoms (Associates of the California Poison Control Center, Poisoning and Drug Overdose, pp. 142-143. 5th Ed). We present the first reported case of a man with delayed presentation to the emergency department after ingesting 24 g of caffeine in a suicide attempt who suffered complications of severe rhabdomyolysis and acute renal failure requiring subsequent hemodialysis.

Door-to-balloon times from freestanding emergency departments meet ST-segment elevation myocardial infarction reperfusion guidelines.

BACKGROUND: Freestanding emergency departments (FEDs) introduce a challenge to physicians who care for the patient with an ST-segment elevation myocardial infarction (STEMI) because treatment is highly time dependent. FEDs have no percutaneous coronary intervention (PCI) capabilities, which necessitates transfer to a PCI-capable facility or fibrinolysis. STUDY OBJECTIVE: Our aim was to determine the proportion of STEMI patients who arrived to an FED and were subsequently transferred for PCI and met the door-to-balloon reperfusion guidelines of 90 min. METHODS: This was a dual-center retrospective cohort review of all patients 18 years and older who were diagnosed with an STEMI and presented to the main hospital-affiliated FEDs. Electronic medical records and emergency medical services documentation were reviewed for all cases since the opening of the FEDs in July 2007 and August 2009, respectively. Key time points were abstracted and statistical evaluation was performed using Fisher's exact test. RESULTS: A total of 47 patients met inclusion criteria. Median door-to-transport time was 34 min (interquartile range [IQR] 15 min). Median transport time from the FEDs to the main hospital catheterization laboratory was 21 min (IQR 5 min). Median arrival at the catheterization laboratory-to-balloon time was 25 min (IQR 13 min). Median total door-to-balloon time was 83 min (IQR 10.5 min), with 78.7% meeting the American Heart Association's recommended guidelines of ≤ 90 min. CONCLUSION: STEMI patients initially seen at two FEDs achieved door-to-balloon time goals of < 90 min.

Spontaneous septic arthritis in a patient without trauma, coinfection, or immunosuppression.

Septic arthritis is a rare infection, most often affecting the knee and hip [1]. Infections are often secondary to joint repair or replacement surgery, systemic infection, or intravenous recreational drug use [1,2].Diabetes, rheumatoid arthritis, hepatic dysfunction, and immunosuppression are common risk factors [1,2]. Although septic arthritis can occur spontaneously, such occurrences are rare. We report a case of a previously healthy 54-year-old woman with no known risk factors presenting to a freestanding emergency department with 5 days of shoulder pain.

Posted emergency department wait times are not always accurate.

OBJECTIVES: Hospitals around the United States are advertising emergency department (ED) wait times. The objective was to measure the difference between publicly posted and actual ED wait times and to compare these between ED site volumes. METHODS: This study was a retrospective consecutive sample of ED patients at one hospital system with four EDs. The wait times of 8,889 patients were included in this analysis. One ED was in a large teaching hospital with 5,000 ED patients per month; the other three were freestanding or community EDs without teaching and with fewer than 2,000 ED patients per month each. The publicly posted ED wait times at the time of patient arrival were recorded and compared to the actual wait times as retrieved from the ED tracking system. The difference between posted and actual wait times for each site was calculated. Separate one-way analysis of variance (ANOVA) tests with post hoc testing were conducted to assess actual wait time and wait time difference between ED sites. RESULTS: Mean and standard deviation (SD) wait time difference at the main ED with a volume of 5,000 patients per month was 31.5 (±61.2) minutes. At the facilities with fewer than 2,000 ED patients per month each, the differences in wait times were 4.2 (±21.8), 8.6 (±23.8), and 1.3 (±11.9) minutes. ANOVA results revealed that the main ED had significantly different actual wait time and wait time differences (p < 0.05) when compared to the other three EDs. CONCLUSIONS: In one hospital system, publicly posted ED wait times show better accuracy in EDs that see 2,000 or fewer patients per month and less accuracy for an ED that sees 5,000 patients per month, likely due to flow confounders.