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Objective: Environmental DNA (eDNA) detection is a transformative tool for ecological surveys which in many cases offers greater accuracy and cost-effectiveness for tracking low-density, cryptic species compared to conventional methods. For the use of targeted quantitative PCR (qPCR)-based eDNA detection, protocols typically require freshly prepared reagents for each sample, necessitating systematic evaluation of reagent stability within the functional context of eDNA standard curve preparation and environmental sample evaluation. Herein, we assessed the effects of long-term storage and freeze–thaw cycles on qPCR reagents for eDNA analysis across six assays. Results: Results demonstrate qPCR plates (containing pre-made PCR mix, primer-probe, and DNA template) remain stable at 4 °C for three days before thermocycling without fidelity loss irrespective of qPCR assay used. Primer-probe mixes remain stable for five months of − 20 °C storage with monthly freeze-thaw cycles also irrespective of qPCR assay used. Synthetic DNA stocks maintain consistency in standard curves and sensitivity for three months under the same conditions. These findings enhance our comprehension of qPCR reagent stability, facilitating streamlined eDNA workflows by minimizing repetitive reagent preparations. [ABSTRACT FROM AUTHOR]

A current challenge for environmental DNA (eDNA) applications is how to account for an environmental (or false-positive) background in surveys. We performed two controlled experiments in the Goldstream Hatchery in British Columbia using a validated coho salmon (Oncorhynchus kisutch) eDNA assay (eONKI4). In the density experiment at high copy number, eDNA in 2 L water samples was measured from four 10 kL tanks containing 1 to 65 juvenile coho salmon. At these densities, we obtained a strong positive 1:1 relationship between predicted copy number/L and coho salmon biomass (g/L). The dilution experiment simulated a situation where fish leave a pool environment, and water from upstream continues to flow through at rates of 141–159 L/min. Here, three coho salmon were placed in four 10 kL tanks, removed after nine days, and the amount of remaining eDNA was measured at times coinciding with dilutions of 20, 40, 80, 160, and 1000 kL. The dilution experiment demonstrates a novel method using Binomial–Poisson distributions to detect target species eDNA at low copy number in the presence of an environmental background. This includes determination of the limit of blank with background (LOB-B) with a controlled false positive rate, and limit of detection with background (LOD-B) with a controlled false negative rate, which provides a statistically robust "Detect" or "No Detect" assessment for eDNA surveys. [ABSTRACT FROM AUTHOR]

The coastal tailed frog (Ascaphus truei) is endemic to the Pacific Northwest of North America and is listed as a species of Special Concern under the Canadian Species at Risk Act. Its range is limited to British Columbia where it occurs widely west of the Coast Mountain Ranges extending north almost to the Alaskan Panhandle. The present study focused on surveying within the Cayoosh, Bridge (Shulaps), Seton, Anderson, Carpenter, and Downton Lake drainages. Four years of previous inventory efforts using conventional time-constrained search (TCS) methods detected tailed frog at 23/292 discrete sites (7.9% detection rate) in seven watersheds. Non-invasive environmental DNA (eDNA) methods hold promise for cryptic and low-abundance species detection. We rigorously validated a quantitative real-time polymerase chain reaction (qPCR)-based tool for detecting coastal tailed frog eDNA in water samples. This eASTR4 test is highly specific and sensitive. We applied a two-step targeted eDNA analysis approach on duplicate filtered water samples from a total of 72 sites collected over five days. The first IntegritE-DNA step mitigates false negative results and tests all DNA samples for the ability to support amplification from endogenous plant chloroplast DNA as a measure of sample viability. Three DNA samples failed this step even after inhibitor clean up suggesting that these samples were poor quality and not reliable for targeted species’ DNA analyses. All other DNA samples were deemed viable and were then tested for species-specific DNA. Coastal tailed frog eDNA was detected in 55/72 (76%) discrete stream reaches; nine sites with historical known occurrence were all eDNA positive. The false negative rate for TCS compared to eDNA methods was 58%. The results expand known coastal tailed frog distribution to 24 watersheds effectively more than tripling extant occurrences and confirm a previously suspected, apparently isolated coastal tailed frog metapopulation in the Shulaps drainage. [ABSTRACT FROM AUTHOR]

Several magnetic resonance parallel imaging techniques require explicit estimates of the receive coil sensitivity profiles. These estimates must be accurate over both the object and its surrounding regions to avoid generating artifacts in the reconstructed images. Regularized estimation methods that involve minimizing a cost function containing both a data-fit term and a regularization term provide robust sensitivity estimates. However, these methods can be computationally expensive when dealing with large problems. In this paper, we propose an iterative algorithm based on variable splitting and the augmented Lagrangian method that estimates the coil sensitivity profile by minimizing a quadratic cost function. Our method, ADMM-Circ, reformulates the finite differencing matrix in the regularization term to enable exact alternating minimization steps. We also present a faster variant of this algorithm using intermediate updating of the associated Lagrange multipliers. Numerical experiments with simulated and real data sets indicate that our proposed method converges approximately twice as fast as the preconditioned conjugate gradient method over the entire field-of-view. These concepts may accelerate other quadratic optimization problems. [ABSTRACT FROM AUTHOR]

A simulated high-strength [chemical oxygen demand (COD) ] wastewater containing primarily acrylic acid, acetic acid, and formaldehyde was treated in an anaerobic membrane bioreactor. The reactor consisted of a 25 L complete-mix tank fitted with three Kubota flat-panel membrane cartridges. The reactor was operated at an organic loading rate of . More than 99% removal of COD and formaldehyde was achieved. Acrylic acid concentration in the effluent stream was below the detection limit. This preliminary study, which lasted for 45 days, showed the potential of application of anaerobic membrane bioreactors in treatment of high-strength wastewaters containing toxic substances like acrylic acid and formaldehyde. [ABSTRACT FROM AUTHOR]

Background: Chronic obstructive pulmonary disease (COPD) is increasing worldwide and thus its associated morbidity and mortality. However, COPD often goes undiagnosed. Objectives: We evaluated the rate of non-diagnosed irreversible airway obstruction (AO) and characterized this patient group. We further assessed the possible effects of conducting targeted spirometry in a population sample in Salzburg, Austria, as part of the Burden of Obstructive Lung Disease (BOLD) study. Methods: 1,258 adults ≥40 years of age completed a questionnaire and performed spirometry before and after bronchodilator therapy (post-BD). Irreversible AO was defined as post-BD FEV1/FVC below the lower limit of normal; we used the FEV1% predicted (pred.) to further grade the disease. Participants without a physician diagnosis of COPD who reported respiratory symptoms and a history of risk factors (ever smoking or occupational risk) were defined as eligible for targeted spirometry. Results: 85.9% (171/199) of the participants with irreversible AO did not report a prior diagnosis of COPD. Non-diagnosed AO was inversely related to severity, age, self-reported prior respiratory diseases and cough as a respiratory symptom. 343 participants were eligible for targeted spirometry and irreversible AO was present in 86 (25.1%) participants. Therefore, targeted spirometry could reduce the underdiagnosis of irreversible AO of any severity by 50.3% (86 of 171). The diagnosis of 1 person with FEV1 <80% pred. would require spirometry in 8.4 subjects (95% confidence interval 6.2-11.1). Conclusion: Although several factors are associated with non-diagnosed AO, spirometry in individuals with respiratory symptoms and exposure to risk factors could reduce undiagnosed irreversible AO by half. Copyright © 2010 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

[Display omitted] • Cryptic, fossorial snake distribution is challenging to determine. • Environmental DNA (eDNA) may solve this but there are few terrestrial applications. • A new sharp-tailed snake eDNA assay was developed and validated. • eDNA was measured from swabs and soil under artificial cover objects. • Detection rate was an order of magnitude better than conventional surveys. Performing ecological surveys for secretive, fossorial snakes is challenging. Traditional survey methods involve visual observation under artificial cover objects (ACOs); this is labor-intensive and requires multiple consistent surveys of suitable habitats. Detection of snake DNA deposited under ACOs represents an innovative method for species detection. However, for terrestrial species, common issues with soil-based methods include the challenges of adequately removing enzyme inhibitors that reduce environmental DNA (eDNA) detection and potential photodegradation of DNA taken from surface samples. These issues may be circumvented by obtaining swabs and soil samples directly from the underside of ACOs for eDNA analysis. We demonstrate the application of this method in surveys of sharp-tailed snake (Contia tenuis), an endangered species under the Canadian Species at Risk Act. We describe the design and validation of a new quantitative real-time polymerase chain reaction (qPCR)-based eDNA eCOTE3 assay with high specificity and sensitivity for sharp-tailed snake. We developed a practical and robust protocol for obtaining eDNA samples by swabbing the underside of ACOs and collecting soil samples under ACOs. Traditional surveys were conducted over two successive years (2018–19) on 220 paired ACOs at 110 sites monitored between 12 and 30 times each. Of the 6,060 ACO visits, only 24 resulted in sharp-tailed snake observations (0.4% success rate) illustrating the considerable difficulty in detecting these snakes. During this same time, 109 swabs were taken directly from the undersides of ACOs and 78 soil samples were collected from a subset of these ACOs. Of the 24 occurrences where sharp-tailed snakes were visually observed, 13 of 23 ACO swabs (57%) and nine of 20 soil samples (45%) tested positive for DNA. eDNA deposition is likely low because of the small size and behavior of this cryptic species, yet DNA was detected from soil exposed to captured snakes for only 10 min. Nevertheless, sharp-tailed snake eDNA was detected at eight sites (9%) from ACO swabs (n = 86) and seven sites (13%) from soil samples (n = 56) where snakes were not observed. This is an overall detection rate of 25% (14/56) for swab and soil samples testing positive in sites where both were tested, representing a substantial reduction in the effort required for detection of this species. Given the time-consuming nature of traditional surveys, eDNA holds great promise as a complementary survey tool for this terrestrial species. While further work is needed to delineate DNA deposition rates, this work represents a significant advance in monitoring a challenging species. [ABSTRACT FROM AUTHOR]

• Forage fish are keystones of the coastal marine food web. • Pacific Sand Lance and Surf Smelt habitat use data are difficult to obtain. • Environmental DNA (eDNA) may improve data from visual egg surveys from sand samples. • Novel eDNA assays for both species were developed and validated. • Detection rate shows promise for broad application of eDNA for forage fish. Many coastal forage fish species spend most of their life history using the water column, but some species such as Pacific Sand Lance (Ammodytes personatus) and Surf Smelt (Hypomesus pretiosus) have a requirement for intertidal sediments for spawning and or burying. Detection of suitable but typically uncommon intertidal habitats for spawning has historically relied on extensive visual surveys for eggs from large sand samples. In the present study, we developed and validated two new non-destructive qPCR-based tools for detecting Sand Lance and Surf Smelt eDNA from small sand samples. A total of 279 composite transect sand samples were collected from 101 beaches and were tested for eDNA sample integrity using the IntegritE-DNATM test. Initial testing showed that all but 42 of 46 samples passed after an inhibitor clean-up step. Of the 101 beaches sampled 275 times, there were 111 detections of Sand Lance on 52 beaches, and there were 13 detections of Surf Smelt on 10 beaches. One hundred and eighty-one samples were paired with visual egg counts and 76 (42%) of these tested positive for Sand Lance while 9 of 132 (7%) samples tested positive for Surf Smelt. For the 27 paired samples with two or more Sand Lance eggs visually detected there was a significant positive correlation (r = 0.694, p = 0.0001) with eDNA copies per liter suggesting that a larger Sand Lance presence is easily and reliably detected using eDNA. The methods and approaches described herein will serve as a foundation for broad application of eDNA approaches for forage fish species that utilize intertidal benthic habitats. [ABSTRACT FROM AUTHOR]

Objective: To determine the possible association between insomnia and risk of type 2 diabetes mellitus (T2DM) in the naturalistic clinical setting., Research Design and Methods: We conducted a retrospective cohort study to examine the risk of developing T2DM among patients with pre-diabetes with and without insomnia. Participants with pre-diabetes (identified by a physician or via two laboratory tests) between January 1, 2007 and December 31, 2015 and without sleep apnea were followed until December 31, 2016. Patients were determined to have T2DM when two of the following occurred within a 2-year window: physician-entered outpatient T2DM diagnosis (International Classification of Diseases [ICD]-9 250.00; ICD-10 E11), dispensing of an antihyperglycemia agent, and hemoglobin A1c (A1c) >6.5% (48 mmol/mol) or fasting plasma glucose (FPG) >125 mg/dL. One hospital inpatient stay with an associated T2DM diagnosis was also sufficient for classification of T2DM., Results: Our cohort consisted of 81 233 persons with pre-diabetes, 24 146 (29.7%) of whom had insomnia at some point during the 4.3-year average observation period. After adjustment for traditional risk factors, those with insomnia were 28% more likely to develop T2DM than those without insomnia (HR 1.28; 95% CI 1.24 to 1.33). The estimate was essentially unchanged after adjusting for baseline A1c level (HR 1.32; 95% CI 1.25 to 1.40) or FPG (HR 1.28; 95% CI 1.23 to 1.33)., Conclusions: Insomnia imparts an increased risk of T2DM comparable with that conferred by traditional risk factors (eg, overweight, non-white race, cardiovascular risk factors). This association could have clinical importance because it suggests a new potentially modifiable risk factor that could be targeted to prevent diabetes., Competing Interests: Competing interests: ESL has had unrelated funding from Merck in the last years. GAN has unrelated grant funding from Boehringer-Ingelheim, Merck & Co, Sanofi, Janssen Pharmaceuticals and Amarin.

Most information concerning possible cancer risks attributable to lifetime exposure to diagnostic x-rays comes from studies in which x-ray history was ascertained by interview or questionnaire, but little is known about the accuracy of such information. The authors assessed agreement between medical x-ray histories obtained through interview and by review of medical records from thyroid cancer case-control studies conducted in Sweden (1985-1992; 123 cases and 123 controls) and from members of a prepaid health plan in the United States (1986-1991; 50 cases and 50 controls). In both studies, substantial disagreement was found between the numbers of x-ray examinations reported in the interview and in the medical records. There was an indication of relatively poorer reporting among controls, particularly for certain types of x-ray examinations and for large numbers of such examinations. Estimates of the risk associated with exposure to diagnostic x-rays were similar, regardless of whether interview or medical record data were used, even though ordinal dose classifications based on the two sources differed considerably. In populations with a high frequency of exposure, spurious associations with numbers of x-ray examinations or estimated thyroid dose might arise because of differences in recall. However, in the present data, reporting errors by cases and controls seemed to be largely nondifferential.