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At least 451 records · Page 25

Large-scale survey of prion protein genetic variability in scrapie disease-free goats from the United States

Scrapie is a slowly progressive neurodegenerative disease of small ruminants caused by an accumulation of an abnormal isoform of prion protein in the central nervous system. Polymorphisms of the prion protein gene (PRNP) strongly modulate scrapie resistance and incubation period in goats. The aim of this study was to identify PRNP genetic variability in goats across the United States. Blood from a total of 6,029 apparent scrapie disease-free goats from 654 operations and 19 breeds were analyzed. Sequencing of PRNP revealed 26 genotypes with different rates based on eight codons. The GG127, RR154, and QQ222 genotypes were predominant and showed a remarkably high rate across all goats. The QK222 and NS146 genotypes, known to be protective against scrapie, were found in 0.6% [with 95% CI = (0.3, 1.2)] and 22.0% [95% CI = (19.1, 25.2)] of goats, respectively. The QK222 genotype was found in 23.1% of Oberhasli goats tested, with 95%CI = (3.9, 68.7)] and 22.0% of Toggenburg goats tested with 95%CI = (9.7, 42.5)], while NS146 was found in 65.5% of Savannah goats tested, with 95%CI = (30.8, 89.9), 36.7% of Boer goats tested, with 95%CI = (33.1, 40.4), 36.3% of Nubian goats tested, with 95%CI = (27.0, 46.7)], and 35.6% of LaMancha goats tested, with 95%CI = (22.8, 50.8%). The MM142 and IM142 genotypes were found more frequently in goats on dairy operations, while the HR143, NS146, and ND146 genotypes were found more frequently in goats on meat operations. Goats in the east region had a higher percentage of goats with RH154, RQ211, and QK222 genotypes than goats in the west region. The results of this study showed high genetic variability of PRNP among the U.S. goat population, with differences by location and breed, and may serve as a rationale for development of goat breeding programs at the national level to mitigate the risk of scrapie.

59 BASIC BIOLOGICAL SCIENCES↗

Quantile-specific heritability of plasma fibrinogen concentrations

Fibrinogen is a moderately heritable blood protein showing different genetic effects by sex, race, smoking status, pollution exposure, and disease status. These interactions may be explained in part by “quantile-dependent expressivity”, where the effect size of a genetic variant depends upon whether the phenotype (e.g. plasma fibrinogen concentration) is high or low relative to its distribution. Determine whether fibrinogen heritability (h 2 ) is quantile-specific, and whether quantile-specific h 2 could account for fibrinogen gene-environment interactions. Plasma fibrinogen concentrations from 5689 offspring-parent pairs and 1932 sibships from the Framingham Heart Study were analyzed. Quantile-specific heritability from offspring-parent (β OP , h 2 = 2β OP /(1+r spouse )) and full-sib regression slopes (β FS , h 2 = {(1+8r spouse β FS ) 0.05 –1}/(2r spouse )) were robustly estimated by quantile regression with nonparametric significance assigned from 1000 bootstrap samples. Quantile-specific h 2 (±SE) increased with increasing percentiles of the offspring’s age- and sex-adjusted fibrinogen distribution when estimated from β OP (P trend = 5.5x10 -6 ): 0.30±0.05 at the 10 th , 0.37±0.04 at the 25 th , 0.48±0.05 at the 50 th , 0.61±0.06 at the 75 th , and 0.65±0.08 at the 90 th percentile, and when estimated from β FS (P trend = 0.008): 0.28±0.04 at the 10 th , 0.31±0.04 at the 25 th , 0.36±0.03 at the 50 th , 0.41±0.05 at the 75 th , and 0.50±0.06 at the 90 th percentile. The larger genetic effect at higher average fibrinogen concentrations may contribute to fibrinogen’s greater heritability in women than men and in Blacks than Whites, and greater increase from smoking and air pollution for the FGB -455G>A A-allele. It may also explain greater fibrinogen differences between: 1) FGB -455G>A genotypes during acute phase reactions than usual conditions, 2) GTSM1 and IL-6 -572C>G genotypes in smokers than nonsmokers, 3) FGB -148C>T genotypes in untreated than treated diabetics, and LPL PvuII genotypes in macroalbuminuric than normoalbuminuric patients. Fibrinogen heritability is quantile specific, which may explain or contribute to its gene-environment interactions. The analyses do not disprove the traditional gene-environment interpretations of these examples, rather quantile-dependent expressivity provides an alternative explanation that warrants consideration.

59 BASIC BIOLOGICAL SCIENCES↗

Comparative evaluation of antimicrobial activity of human granulysin, bovine and porcine NK-lysins against Shiga toxin-producing Escherichia coli O157:H7

Shiga toxin-producing Escherichia coli (STEC) O157:H7 (O157) is a foodborne pathogen causing human disease ranging from hemorrhagic colitis and hemolytic uremic syndrome to kidney failure, while remaining harmless to cattle, its primary reservoir. The severity of the human disease associated mainly with Shiga toxin production and a global emergence of antibiotic resistant STEC highlights the need for effective non-antibiotic, pre-harvest strategies to reduce O157 in cattle, the principal source of human infection. Towards this goal three synthetic antimicrobial peptides (AMPs): human granulysin (hGRNL), bovine NK-lysin (bNK2A), and porcine NK-lysin (pNKL), were tested in vitro against O157 isolates. As expected, circular dichroism spectroscopy findings were consistent with a predominantly α-helical conformation for all three AMPs in an environment mimicking bacterial outer surface or liposaccharides. The minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations of hGRNL (200 μM), bNK2A (12.5 μM against strain 86–24 and 25 μM against EDL933), and pNKL (6.25 μM) were determined using the Clinical and Laboratory Standards Institute broth microdilution method in Müeller-Hinton broth (cation-adjusted). The bNK2A and pNKL AMPs did not induce Shiga toxin expression in O157 at MIC, as there was a significant decrease or no change in toxin expression following 4- or 20 h incubation with the AMPs; bNK2A p <0.0001 (4 h) and p = 0.4831 (20 h); pNKL p <0.0001 (4 h) and p = 0.0001 (20 h). Propidium iodide uptake assay revealed faster O157 membrane damage or killing kinetics with bNK2A and pNKL compared to hGRNL. Nonetheless, transmission electron microscopy demonstrated that all three AMPs mediated damage to O157 membranes. In contrast, the three AMPs showed minimal cytotoxicity (<2%) against cattle red blood cells at tested concentrations (0.39–50 μM). Overall, our results demonstrate the potential for bNK2A and pNKL to be further developed into novel non-antibiotic agents to reduce O157 shedding in cattle.

59 BASIC BIOLOGICAL SCIENCES↗

Temporal serum neurofilament light chain concentrations in sheep inoculated with the agent of classical scrapie

Objective: Neurofilament light chain (Nf-L) has been used to detect neuroaxonal damage in the brain caused by physical injury or disease. The purpose of this study was to determine if serum Nf-L could be used as a biomarker for pre-symptomatic detection of scrapie in sheep. Methods: Four sheep with prion protein genotype AVQQ were intranasally inoculated with the classical scrapie strain x124. Blood was collected every 4 weeks until 44 weeks post-inoculation, at which point weekly collection commenced. Serum was analyzed using single molecule array (Quanterix SR-X) to evaluate Nf-L concentrations. Results: Scrapie was confirmed in each sheep by testing homogenized brainstem at the level of the obex with a commercially available enzyme immunoassay. Increased serum Nf-L concentrations were identified above the determined cutoff during the last tenth of the respective incubation period for each sheep. Throughout the time course study, PrP Sc accumulation was not detected antemortem by immunohistochemistry in rectal tissue at any timepoint for any sheep. RT-QuIC results were inconsistently positive throughout the timepoints tested for each sheep; however, each sheep had at least one timepoint detected positive. When assessing serum Nf-L utility using receiver operator characteristic curves against different clinical parameters, such as asymptomatic and symptomatic (pruritus or neurologic signs), results showed that Nf-L was most useful at being an indicator of disease only late in disease progression when neurologic signs were present. Conclusion: Serum Nf-L concentrations in the cohort of sheep increased as disease progressed; however, serum Nf-L did not increase during the presymptomatic window. The levels increased substantially throughout the final 10% of the animals’ scrapie incubation period when other clinical signs were present. Serum Nf-L is not a reliable biomarker for pre-clinical detection of scrapie.

59 BASIC BIOLOGICAL SCIENCES↗

Binding of adenovirus species C hexon to prothrombin and the influence of hexon on vector properties in vitro and in vivo

The majority of adenovirus (Ad) vectors are based on human Ad type 5, which is a member of Ad species C. Species C also includes the closely-related types 1, 2, 6, 57 and 89. It is known that coagulation factors bind to Ad5 hexon and play a key role in the liver tropism of Ad5 vectors, but it is unclear how coagulation factors affect vectors derived from other species C Ads. We evaluated species C Ad vectors both in vitro and following intravenous injection in mice. To assess the impact of hexon differences, we constructed chimeric Ad5 vectors that contain the hexon hypervariable regions from other species C types, including vectors with hexon mutations that decreased coagulation factor binding. After intravenous injection into mice, vectors with Ad5 or Ad6 hexon had strong liver tropism, while vectors with chimeric hexon from other Ad types had weaker liver tropism due to inhibition by natural antibodies and complement. In addition, we discovered a novel ability of hexon to bind prothrombin, which is the most abundant coagulation factor in blood, and we found striking differences in the affinity of Ads for human, mouse and bovine coagulation factors. When compared to Ad5, vectors with non-Ad5 species C hexons had considerably higher affinity for both human and mouse prothrombin. Most of the vectors tested were strongly dependent on coagulation factors for liver transduction, but vectors with chimeric Ad6 hexon showed much less dependence on coagulation factors than other vectors. We found that in vitro neutralization experiments with mouse serum predicted in vivo behavior of Ad5 vectors, but in vitro experiments did not predict the in vivo behavior of vectors based on other Ad types. In sum, hexons from different human Ad species C viruses confer diverse properties on vectors, including differing abilities to target the liver.

Tian, Jie↗

Ketone monoester plus high‐dose glucose supplementation before exercise does not affect immediate post‐exercise erythropoietin concentrations versus glucose alone

Abstract The objective of this study was to examine the effect of consuming ketone monoester plus a high dose of carbohydrate from glucose (KE + CHO) on the change in erythropoietin (EPO) concentrations during load carriage exercise compared with carbohydrate (CHO) alone. Using a randomized, crossover design, 12 males consumed KE + CHO (573 mg KE/kg body mass, 110 g glucose) or CHO (110 g glucose) 30 min before 4 miles of self‐paced treadmill exercise (KE + CHO:51 ± 13%, CHO: 52 ± 12% V̇O 2peak ) wearing a weighted vest (30% body mass; 25 ± 3 kg). Blood samples for analysis were obtained under resting fasted conditions before (Baseline) consuming the KE + CHO or CHO supplement and immediately after exercise (Post). βHB increased ( p < 0.05) from Baseline to Post in KE + CHO, with no change in CHO. Glucose and glycerol increased ( p < 0.05) from Baseline to Post in CHO, with no effect of time in KE + CHO. Insulin and lactate increased ( p < 0.05) from Baseline to Post independent of treatment. EPO increased ( p < 0.05) from Baseline to Post in KE + CHO and CHO with no difference between treatments. Although KE + CHO altered βHB, glucose, and glycerol concentrations, results from this study suggest that KE + CHO supplementation before load carriage exercise does not enhance immediate post‐exercise increases in EPO compared with CHO alone.

Howard, Emily E.↗

Chymotrypsin-like Elastase-1 Mediates Progressive Emphysema in Alpha-1 Antitrypsin Deficiency

Alpha-1 antitrypsin (AAT) deficiency is a rare disease affecting approximately 1 in 2000 White individuals with approximately 10% of these individuals developing AATD lung disease. This lung disease is marked by progressive alveolar loss despite the withdrawal of triggering agents such as cigarette smoke. Although the PiZZ genotype is the most common mutation, there are over 100 described variants making true population estimates difficult and AAT deficiency is typically diagnosed by reduced levels of AAT in the blood. Typically developing in the fourth and fifth decades of life, AAT-deficient lung disease is marked by progressive emphysema and is the fourth leading indication for lung transplantation. AAT augmentation therapy does not prevent disease progression making the development of new therapeutic approaches critical. Chymotrypsin-like elastase 1 (CELA1) is a serine protease synthesized and secreted by alveolar type 2 cells with a physiologic role in reducing postnatal lung elastance. At a molecular level, CELA1 binds and cleaves non-crosslinked, hydrophobic domains of tropoelastin, and its binding to lung elastin fibers is increased with strain—similar to other pancreatic elastases. CELA1 is neutralized by covalent binding with AAT, and Cela1 -/- mice were completely protected from emphysema in an antisense oligonucleotide model of AAT-deficient emphysema. This model, however, did not include any injury apart from administration of the antisense oligonucleotide with levels of emphysema exceeding that seen in mice with genetic ablation of 5 Serpina1 paralogues and subjected to tracheal lipopolysacharide or cigarette smoke. Here, we use this murine genetic model of AAT deficiency to test the role of the CELA1 gene in AAT-deficient emphysema using multiple models to show that CELA1 has a role in progressive airspace enlargement in AAT-deficiency independent of inflammation.

60 APPLIED LIFE SCIENCES↗

A Million Person Study Innovation: Evaluating Cognitive Impairment and other Morbidity Outcomes from Chronic Radiation Exposure Through Linkages with the Centers for Medicaid and Medicare Services Assessment and Claims Data

Here, the study of One Million U.S. Radiation Workers and Veterans, the Million Person Study (MPS), examines the health consequences, both cancer and non-cancer, of exposure to ionizing radiation received gradually over time. Recently the MPS has focused on mortality patterns from neurological and behavioral conditions, e.g., Parkinson's disease, Alzheimer's disease, dementia, and motor neuron disease such as amyotrophic lateral sclerosis. A fuller picture of radiation-related late effects comes from studying both mortality and the occurrence (incidence) of conditions not leading to death. Accordingly, the MPS is identifying neurocognitive diagnoses from fee-for-service insurance claims from the Centers for Medicare and Medicaid Services (CMS), among Medicare beneficiaries beginning in 1999 (the earliest date claims data are available). Linkages to date have identified ∼540,000 workers with available health information. Such linkages provide individual information on important co-factor and confounding variables such as smoking, alcohol consumption, blood pressure, obesity, diabetes and many other health and demographic characteristics. The total person-level set of time-dependent variables, outcomes, organ-specific dose measures, co-factors, and demographics will be massive and much too large to be evaluated with standard software. Thus, development of specialized open-source software designed for large datasets (Colossus) is nearly complete. The wealth of information available from CMS claims data, coupled with individual dose reconstructions, will thus greatly enhance the quality and precision of health evaluations for this new field of low-dose radiation and neurocognitive effects.

Dauer, Lawrence T.↗

Identification of a Class of WNK Isoform-Specific Inhibitors Through High-Throughput Screening

Introduction: WNK [with no lysine (K)] kinases are serine/threonine kinases associated with familial hyperkalemic hypertension (FHHt). WNKs are therapeutic targets for blood pressure regulation, stroke and several cancers including triple negative breast cancer and glioblastoma. Here, we searched for and characterized novel WNK kinase inhibitors. Methods: We used a ~210,000-compound library in a high-throughput screen, re-acquisition and assay, commercial specificity screens and crystallography to identify WNK-isoform-selective inhibitors. Results: We identified five classes of compounds that inhibit the kinase activity of WNK1: quinoline compounds, halo-sulfones, cyclopropane-containing thiazoles, piperazine-containing compounds, and nitrophenol-derived compounds. The compounds are strongly pan-WNK selective, inhibiting all four WNK isoforms. A class of quinoline compounds was identified that further shows selectivity among the WNK isoforms, being more potent toward WNK3 than WNK1. The crystal structure of the quinoline-derived SW120619 bound to the kinase domain of WNK3 reveals active site binding, and comparison to the WNK1 structure reveals the potential origin of isoform specificity. Discussion: The newly discovered classes of compounds may be starting points for generating pharmacological tools and potential drugs treating hypertension and cancer.

60 APPLIED LIFE SCIENCES↗

COVID-19 biomarkers based on respiratory microbiome content

COVID-19 patient care management would greatly benefit from new tools that enable accurate assessment of disease severity and stage, potentially enabling a personalized medicine approach. Detection of the SARS-CoV-2 virus itself, or even quantitation of viral loads, is not sufficient for accurate assessment of disease state beyond diagnosis of infection [eg, doi:10.1093/cid/ciaa344]. Levels of usual suspect protein biomarkers associated with host response to infection [eg, C-reactive protein (CRP); cytokines like IL-6, TNF-alpha, and IL-10; complement proteins like C3a and C5a], and of individual blood cell types (eg, leukocytes, lymphocytes, and subsets thereof), show limited correlation with disease severity and stage, with high patient-to-patient and study-to-study variability [eg, doi:10.1093/cid/ciaa248]. High-dimensional panels of biomarkers should have greater predictive power and resilience to unavoidable sources of variability; however, their assembly from proteins and cell types is extremely difficult, due to technical limitations in analyte measurement, especially with regard to starting material requirements and detection sensitivity. Host response profiling through Next Generation Sequencing (NGS) of gene expression patterns (ie, RNA-Seq) is a promising approach, but at the time of this project there were only two publicly available datasets of relevance [doi:10.1093/cid/ciaa203, doi:10.1080/22221751.2020.1747363], and close inspection of them revealed that each had at least one major flaw that severely undermined its value in supporting robust analysis of host response to SARS-CoV- 2 infection. However, the first of these studies [doi:10.1093/cid/ciaa203] fortuitously collected NGS data not only from host cells, but also from bacteria present in bronchoalveolar lavage fluid (BALF) recovered from COVID-19 patients; and because the respiratory microbiome (in terms of bacterial species content) is far less complex than the human transcriptome, the NGS data collected were sufficient to provide coverage depth supporting robust analysis. Surprisingly, the authors of the study did not carry out a detailed analysis of these data and their potential for revealing important new information about COVID-19. Therefore, we carried out a meta-analysis of the dataset as a first step in evaluating the potential for profiling of respiratory microbiome dynamics as a means of accurately assessing COVID-19 disease state.

59 BASIC BIOLOGICAL SCIENCES↗

Testing a novel peptide drug towards a goal of reducing mortality in critically ill COVID19 patients

COVID 19 has been devastating the globe over the past year, and although our knowledge of how the disease progresses is constantly evolving, there is a significant amount we now about the mechanisms underlying COVID19. There is growing evidence that a 'cytokine storm' is a significant driver of mortality associated with COVID19, with studies showing high levels of hallmark inflammatory indicators in critically ill COVID19 patients. This essentially consists of the patient's immune system going awry and causing white blood cells to constantly release a large number of small molecules called cytokines. The result of this is further activation of the immune system, which ends up attacking not only infected but also healthy patient tissues, resulting in organ failure.

59 BASIC BIOLOGICAL SCIENCES↗

Radiobiology, Omics and Microdosimetry of Systemic and Targeted Radiotherapeutics Workshop

The Radiobiology, Omics and Microdosimetry of Systemic and Targeted Radiotherapeutics Workshop fostered engagement across disciplines on critical topics that impact the development of targeted radionuclide therapies, bringing together scientific and medical professionals actively involved in radiotherapeutic design, optimization testing, patient treatment, and clinical trials along with basic science researchers, whose novel techniques have the potential to further our understanding of targeted radionuclide therapy. Discussion of the applications of radiotherapeutics to pediatric and adult, brain, prostate and blood cancers, and metastatic disease allowed scientific research to be presented in the context of real-world medical applications and provided an important backdrop to the basic research performed in computers, laboratories, and animal facilities. The workshop format, presentations, and panel discussions were designed to consider a holistic approach to the development of systemic and targeted radiotherapies, integrating radiobiology and microdosimetry, biological assessments on the microscale, and physiologically relevant models of tumor and tissue microenvironments. Important needs highlighted in the workshop include better models to assess the biological effects of radionuclide therapy, novel strategies to target radiotherapeutics to disease sites with high specificity, and new dosimetry techniques that address the importance of tumor microenvironments.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

Capability: Selective Endotoxin Uptake

Researchers at Los Alamos National Lab are developing a new capability to selectively absorb lipopolysaccharide, also known as endotoxin. Current sepsis treatment technologies generally target the bacteria causing the infection, but do not mitigate the host response that causes sickness. This disruptive capability could enable the removal of endotoxin from patient blood adding a much needed tool to the toolkit of sepsis treatments. We are seeking a CRADA partner with the desire to develop sepsis treatments and collaborate on the next steps.

59 BASIC BIOLOGICAL SCIENCES↗

Health Management Clinic Report FY18

The Health Management Clinic (HMC) is a worksite specialty clinic designed to provide an exceptional level of health care for Sandia employees with diabetes, cholesterol and blood pressure disorders, and for those employees that need help with smoking cessation, depression, anxiety, sleep disorders, or weight management. With a unified commitment to the best care practices available, the HMC is Sandia’s interface to workplace healthcare and health plan services. The HMC provides Sandia employees access to onsite screenings, health care exams, preventative health education, disease management education, care management, periodic laboratory testing, immunizations, podiatry services, and behavioral, fitness, and nutrition counseling/education. Our multidisciplinary team of health professionals consists of physicians, nurses, medical assistants, certified diabetes educators, dietitians, health educators, and exercise specialists. Services offered by the Health Management clinic have been designed to reduce further complications from disease states and promote healthy behavior changes for Sandia employees.

60 APPLIED LIFE SCIENCES↗

Utilizing Additive Manufacturing for Biomimetic Designs

Chronic kidney disease effects 15% of adult Americans. Current technology utilizes polymer membranes for separation of toxins and waste from the blood. Yet there still is room for development of new materials to improve anti-biofouling characteristics and able to filter a range of molecular weights. This short review describes nano-porous membranes for molecular separation as well as limitations with current membrane designs.

36 MATERIALS SCIENCE↗

Stress Birth and Death: Disruptive Computational Mechanics and Novel Diagnostics for Fluid-to-Solid Transitions

Many materials of interest to Sandia transition from fluid to solid or have regions of both phases coexisting simultaneously. Currently there are, unfortunately, no material models that can accurately predict this material response. This is relevant to applications that "birth stress" related to geoscience, nuclear safety, manufacturing, energy production and bioscience. Accurately capturing solidification and residual stress enables fully predictive simulations of the evolving front shape or final product. Accurately resolving flow of proppants or blood could reduce environmental impact or lead to better treatments for heart attacks, thrombosis, or aneurism. We will address a science question in this proposal: When does residual stress develop during the critical transition from liquid to solid and how does it affect material deformation? Our hypothesis is that these early phases of stress development are critical to predictive simulation of material performance, net shape, and aging. In this project, we use advanced constitutive models with yield stress to represent both fluid and solid behavior simultaneously. The report provides an abbreviated description of the results from our LDRD "Stress Birth and Death: Disruptive Computational Mechanics and Novel Diagnostics for Fluid-to-Solid Transitions," since we have written four papers that document the work in detail and which we reference. We give highlights of the work and describe the gravitationally driven flow visualization experiment on a model yield stress fluid, Carbopol, at various concentrations and flow rates. We were able to collapse the data on a single master curve by showing it was self-similar. We also describe the Carbopol rheology and the constitutive equations of interest including the Bingham-Carreau-Yasuda model, the Saramito model, and the HB-Saramito model including parameter estimation for the shear and oscillatory rheology. We present several computational models including the 3D moving mesh simulations of both the Saramito models and Bingham-Carreau-Yasuda (BCY) model. We also show results from the BCY model using a 3D level set method and two different ways of handling reduced order Hele-Shaw modeling for generalized Newtonian fluids. We present some first ever two-dimensional results for the modified Jeffries Kamani-Donley-Rogers constitutive equation developed during this project. We include some recent results with a successful Saramito-level set coupling that allows us to tackle problems with complex geometries like mold filling in a thin gap with an obstacle, without the need for remeshing or remapping. We report on some experiments for curing systems where fluorescent particles are used to track material flow. These experiments were carried out in an oven on Sylgard 184 as a model polymerizing system. We conclude the report with a summary of accomplishments and some thoughts on follow-on work.

36 MATERIALS SCIENCE↗

International Benchmarking: A Global Perspective of Workplace Drug and Alcohol Testing

Workplace drug and alcohol testing has been widely implemented in diverse industries and is designed to detect the use of and deter the influence of legal and illegal substances that would impair the ability to perform work that has significant security and safety implications. Across the globe, patterns of drug and alcohol use and abuse and the regulations controlling drug and alcohol use vary by location (e.g., regions, countries, etc.). The prevalence of workplace drug and alcohol testing and protection of individuals’ rights in the workplace similarly tend to exhibit contextual differences. With the advancements in drug-testing technologies and the recent issuance of guidance for additional testing matrices (e.g., oral fluid, hair, etc.) by the U.S. Department of Health and Human Services, gaining a global perspective on the current state of drug and alcohol testing practices and regulations is important for leveraging such advancements to improve the effectiveness of drug and alcohol testing. The U.S. Nuclear Regulatory Commission (NRC) tasked the Pacific Northwest National Laboratory (PNNL) fitness for duty (FFD) team to conduct a benchmarking analysis of international drug and alcohol testing regulations and programs to identify similarities and differences in workplace testing programs and the basis for any differences. The benchmarking study focuses on a policy analysis and benchmarking metrics comparison among different countries and regions. The policy analysis includes the following three types of relevant international and national policies relevant to drug and alcohol testing in the nuclear industry: 1. National drug and alcohol policy 2. Workplace drug and alcohol testing 3. FFD regulations for Nuclear Power Plants The international drug and alcohol benchmarking metrics focusses on the use of blood, breath, urine, hair, and oral fluid in testing; drug and drug metabolite cutoff values; point-of-collection testing devices; roadside testing methodologies; innovations in drug testing; and the extent of availability of such data. Gaining an understanding of workplace drug and alcohol testing in other countries as well as information about new testing methods, matrices, tools, and technologies that other countries have adopted provides NRC with international comparative information regarding the technical basis for such programs and methods as it relates to the protection of worker and public health and safety.

98 NUCLEAR DISARMAMENT, SAFEGUARDS, AND PHYSICAL P↗