The structure of NAD+ consuming protein Acinetobacter baumannii TIR domain shows unique kinetics and conformations.

Toll-like and interleukin-1/18 receptor/resistance (TIR) domain-containing proteins function as important signaling and immune regulatory molecules. TIR domain-containing proteins identified in eukaryotic and prokaryotic species also exhibit NAD+ hydrolase activity in select bacteria, plants, and mammalian cells. We report the crystal structure of the Acinetobacter baumannii TIR domain protein (AbTir-TIR) with confirmed NAD+ hydrolysis and map the conformational effects of its interaction with NAD+ using hydrogen-deuterium exchange-mass spectrometry. NAD+ results in mild decreases in deuterium uptake at the dimeric interface. In addition, AbTir-TIR exhibits EX1 kinetics indicative of large cooperative conformational changes, which are slowed down upon substrate binding. Additionally, we have developed label-free imaging using the minimally invasive spectroscopic method 2-photon excitation with fluorescence lifetime imaging, which shows differences in bacteria expressing native and mutant NAD+ hydrolase-inactivated AbTir-TIRE208A protein. Our observations are consistent with substrate-induced conformational changes reported in other TIR model systems with NAD+ hydrolase activity. These studies provide further insight into bacterial TIR protein mechanisms and their varying roles in biology.

Competitive TcO4-, IO3-, and CrO42- Incorporation into Ettringite.

Ettringite is a naturally occurring mineral found in cementitious matrices that is known for its ability to incorporate environmentally mobile oxyanion contaminants. To better assess this immobilization mechanism for contaminants within cementitious waste forms intended for nuclear waste storage, this work explores how mixed oxyanion contaminants compete for ettringite incorporation and influence the evolving mineralogy. Ettringite was precipitated in the presence of TcO4-, IO3-, and/or CrO42-, known contaminants of concern to nuclear waste treatment, over pre-determined precipitation periods. Solution analyses quantified contaminant removal, and the collected solid was characterized using bulk and microprobe X-ray diffraction coupled with pair distribution function and microprobe X-ray fluorescence analyses. Results suggest that ≥96% IO3- is removed from solution, regardless of ettringite precipitation time or the presence of TcO4- or CrO42-. However, TcO4- removal remained <20%, was not significantly improved with longer ettringite precipitation times, and decreased to zero in the presence of IO3-. When IO3- is co-mingled with CrO42-, calcite and gypsum are formed as secondary mineral phases, which allows for oxyanion partitioning, e.g., IO3- incorporation into ettringite, and CrO42- incorporation into calcite. Results from this work exemplify the importance of competitive immobilization when assessing waste form performance and environmental risk of contaminant release.

Hydroxide promotes ion pairing in the NaNO2-NaOH-H2O system.

Nitrite (NO2-) is a prevalent nitrogen oxyanion in environmental and industrial processes, but its behavior in solution, including ion pair formation, is complex. This solution phase complexity impacts industries such as nuclear waste treatment, where NO2- significantly affects the solubility of other constituents present in sodium hydroxide (NaOH)-rich nuclear waste. This work provides molecular scale information into sodium nitrite (NaNO2) and NaOH ion-pairing processes to provide a physical basis for later development of thermodynamic models. Solubility isotherms of NaNO2 in aqueous mixtures with NaOH and total alkalinity were also measured. Spectroscopic characterization of these solutions utilized high-field nuclear magnetic resonance spectroscopy (NMR) and Raman spectroscopy, with additional solution structure detailed by X-ray total scattering pairwise distribution function analysis (X-ray PDF). Despite the NO2- deformation Raman band's insensitivity to added NaOH in saturated NaNO2 solutions, 23Na and 15N NMR studies indicated the Na+ and NO2- chemical environments change likely due to ion pairing. The ion pairing correlates with a decrease in diffusion coefficient of solution species as measured by pulsed field gradient 23Na and 1H NMR. Two-dimensional correlation analyses of the 2800-4000 cm-1 Raman region and X-ray PDF indicated that saturated NaNO2 and NaOH mixtures disrupt the hydrogen network of water into a new structure where the length of the OO correlations is contracted relative to the typical H2O structure. Beyond describing the solubility of NaNO2 in a multicomponent electrolyte mixture, these results also indicate that nitrite exhibits greater ion pairing in mixtures of concentrated NaNO2 and NaOH than in comparable solutions with only NaNO2.

Immobilizing Pertechnetate in Ettringite via Sulfate Substitution.

Technetium-99 immobilization in low-temperature nuclear waste forms often relies on additives that reduce environmentally mobile pertechnetate (TcO4-) to insoluble Tc(IV) species. However, this is a short-lived solution unless reducing conditions are maintained over the hazardous life cycle of radioactive wastes (some ∼10,000 years). Considering recent experimental observations, this work explores how rapid formation of ettringite [Ca6Al2(SO4)3(OH)12·26(H2O)], a common mineral formed in cementitious waste forms, may be used to directly immobilize TcO4-. Results from ab initio molecular dynamics (AIMD) simulations and solid-phase characterization techniques, including synchrotron X-ray absorption, fluorescence, and diffraction methods, support successful incorporation of TcO4- into the ettringite crystal structure via sulfate substitution when synthesized by aqueous precipitation methods. One sulfate and one water are replaced with one TcO4- and one OH- during substitution, where Ca2+-coordinated water near the substitution site is deprotonated to form OH- for charge compensation upon TcO4- substitution. Furthermore, AIMD calculations support favorable TcO4- substitution at the SO42- site in ettringite rather than gypsum (CaSO4·2H2O, formed as a secondary mineral phase) by at least 0.76 eV at 298 K. These results are the first of their kind to suggest that ettringite may contribute to TcO4- immobilization and the overall lifetime performance of cementitious waste forms.

Ion-ion interactions enhance aluminum solubility in alkaline suspensions of nano-gibbsite (α-Al(OH)3) with sodium nitrite/nitrate.

Despite widespread industrial importance, predicting metal solubilities in highly concentrated, multicomponent aqueous solutions is difficult due to poorly understood ion-ion and ion-solvent interactions. Aluminum hydroxide solid phase solubility in concentrated sodium hydroxide (NaOH) solutions is one such case, with major implications for ore refining, as well as processing of radioactive waste stored at U.S. Department of Energy legacy sites, such as the Hanford Site, Washington State. The solubility of gibbsite (α-Al(OH)3) is often not well predicted because other ions affect the activity of hydroxide (OH-) and aluminate (Al(OH)4-) anions. In the present study, we systematically examined the influence of key anions, nitrite (NO2-) and nitrate (NO3-), as sodium salts on the solubility of α-Al(OH)3 in NaOH solutions taking care to establish equilibrium from both under- and oversaturation. Rapid equilibration was enabled by use of a highly pure and crystalline synthetic nano-gibbsite of well-defined particle size and shape. Measured dissolved aluminum concentrations were compared with those predicted by an α-Al(OH)3 solubility model derived for simple Al(OH)4-/OH- systems. Specific anion effects were expressed as an enhancement factor (Alenhc) conveying the excess of dissolved aluminum. At 45 °C, NaNO2 and NaNO3-containing systems exhibited Alenhc values of 2.70 and 1.88, respectively, indicating significant enhancement. The solutions were examined by Raman and high-field 27Al NMR spectroscopy, indicating specific interactions including Al(OH)4--Na+ contact ion pairing and Al(OH)4--NO2-/NO3- ion-ion interactions. Dynamic evolution of the α-Al(OH)3 particles including growth and agglomeration was observed revealing the importance of dissolution/reprecipitation in establishing equilibrium. These studies indicate that incomplete ion hydration, as a result of the low water activity in these concentrated electrolytes, results in: (i) enhanced reactivity of the hydroxide ion with respect to α-Al(OH)3; (ii) increased concentrations of Al(OH)4- in solution; and (iii) stronger ion-ion interactions that act to stabilize the supersaturated solutions. This information on the mechanisms by which α-Al(OH)3 becomes supersaturated is essential for more energy-efficient aluminum processing technologies, including the treatment of millions of gallons of Al(OH)4--rich high-level radioactive waste.

NO to NO2 Conversion Rate Analysis and Implications for Dispersion Model Chemistry Methods using Las Vegas, Nevada Near-Road Field Measurements.

The nitrogen dioxide/oxides of nitrogen (NO2/NOX) ratio is an important surrogate for NO to NO2 chemistry in dispersion models when estimating NOX impacts in a near-road environment. Existing dispersion models use different techniques and assumptions to represent NO to NO2 conversion and do not fully characterize all of the important atmospheric chemical and mechanical processes. Thus, "real-world" ambient measurements must be analyzed to assess the behavior of NO2/NOX ratios near roadways. An examination of NO2/NOX ratio data from a field study conducted in Las Vegas, Nevada (NV), from mid-December, 2008 through mid-December, 2009 provides insights into the appropriateness of assumptions about the NO2/NOX ratio included in dispersion models. Data analysis indicates multiple factors affect the downwind NO2/NOX ratio. These include spatial gradient, background ozone (O3), source emissions of NO and NO2, and background NO2/NOX ratio. Analysis of the NO2/NOX ratio spatial gradient indicates that under high O3 conditions, the change in the ratio is fairly constant once a certain O3 threshold (≥ 30 ppb) is reached. However, under low O3 conditions (< 30 ppb), there are differences between weekdays and weekends, most likely due to a decline in O3 concentrations during the weekday morning hours, reducing the O3 available to titrate the emitted NO, allowing lower NO2/NOX ratios. These results suggest that under high O3 conditions, NOX chemistry is driving the NO2/NOX ratios whereas under low O3 conditions, atmospheric mixing is the driving factor.

Redistribution of heart failure as the cause of death: the Atherosclerosis Risk in Communities Study.

BACKGROUND: Heart failure is sometimes incorrectly listed as the underlying cause of death (UCD) on death certificates, thus compromising the accuracy and comparability of mortality statistics. Statistical redistribution of the UCD has been used to examine the effect of misclassification of the UCD attributed to heart failure, but sex- and race-specific redistribution of deaths on coronary heart disease (CHD) mortality in the United States has not been examined. METHODS: We used coarsened exact matching to infer the UCD of vital records with heart failure as the UCD from 1999 to 2010 for decedents 55 years old and older from states encompassing regions under surveillance by the Atherosclerosis Risk in Communities (ARIC) Study (Maryland, Minnesota, Mississippi, and North Carolina). Records with heart failure as the UCD were matched on decedent characteristics (five-year age groups, sex, race, education, year of death, and state) to records with heart failure listed among the multiple causes of death. Each heart failure death was then redistributed to plausible UCDs proportional to the frequency among matched records. RESULTS: After redistribution the proportion of deaths increased for CHD, chronic obstructive pulmonary disease, diabetes, hypertensive heart disease, and cardiomyopathy, P < 0.001. The percent increase in CHD mortality after redistribution was the highest in Mississippi (12%) and lowest in Maryland (1.6%), with variations by year, race, and sex. Redistribution proportions for CHD were similar to CHD death classification by a panel of expert reviewers in the ARIC study. CONCLUSIONS: Redistribution of ill-defined UCD would improve the accuracy and comparability of mortality statistics used to allocate public health resources and monitor mortality trends.

Complement protein C3 and coronary artery calcium in middle-aged women with polycystic ovary syndrome and controls.

Circulating complement protein C3 (C3) levels have been associated with coronary artery calcification (CAC) in women with systemic lupus erythematosus, but have yet to be evaluated in women with polycystic ovary syndrome (PCOS). We aimed to determine whether C3 levels were elevated in women with PCOS compared to controls and to quantify the association of C3 with cardiovascular disease (CVD) risk factors and CAC and if PCOS modified this association. This cross-sectional analysis included 132 women with PCOS and 155 controls, 35-62 years old, from the third visit of a case-control study. CAC was measured during the study visit, and circulating C3 was measured in stored sera. The presence of CAC and CAC categories (Agatston score 0, 1-9.9 and ≥ 10) were used for logistic and ordinal regression analysis, respectively. C3 levels were not significantly different between women with PCOS and controls. Among all women, C3 was associated with the presence of CAC and increasing CAC groups after adjusting for age, PCOS status and insulin or body mass index (BMI), all p<0.05. In addition, C3 was associated with the presence of CAC after adjusting for age, PCOS status, BMI, insulin and African American race, p=0.049. PCOS status did not modify these associations. In conclusion, circulating C3 levels may prove beneficial in identifying women at risk of CVD in women with PCOS and the general population.

Short-term repeatability of electrocardiographic P wave indices and PR interval.

BACKGROUND: P wave indices and PR interval from 12-lead electrocardiograms (ECGs) are predictors of cardiovascular morbidity and mortality, but their repeatability has not been examined. OBJECTIVES: Determine the short-term repeatability of P wave indices (P axis, maximum P area and duration, P dispersion and P terminal force in V1) and PR interval. METHODS: Participants (n=63) underwent two standard ECGs at each of two visits, two weeks apart. We calculated the intra-class correlation coefficient (ICC), weighted kappa, and minimal detectable change and difference. RESULTS: ICCs were 0.93 for PR interval, 0.78 for P axis, 0.77 for maximum P area, and 0.58 for maximum P duration. Within- and between-visit Kappa were 0.30 and 0.11 for P dispersion, and 0.68 and 0.46 for P terminal force. CONCLUSION: Repeatability of PR duration was excellent, that of P wave axis and maximum area was fair, and maximum P wave duration and terminal force was poor. Repeatability of P wave dispersion was fair within visit, yet poor between visits. These results illustrate potential biases when measurement error of some P wave indices is ignored in clinical and epidemiologic studies.

Short-term repeatability of electrocardiographic Tpeak-Tend and QT intervals.

BACKGROUND: The electrocardiographic (ECG) Tpeak-Tend interval (TpTe) is associated with arrhythmias and sudden cardiac death. TpTe offers a supplementary measure for the QT interval (QT), yet its repeatability has not been established. PURPOSE: Evaluate short-term repeatability of TpTe and QT. METHODS: Four ECGs were obtained on sixty participants. The sources of variation, intra-class correlation coefficient (ICC) - an index of reproducibility - and minimal detectable change (MDC) were estimated for TpTe and QT. The impact of repeated measurements on repeatability was estimated for a hypothetical clinical trial designed to detect drug-induced prolongation of TpTe and QT. RESULTS: We used heart rate-adjusted QT [(QT)a] but TpTe in the study group was rate-invariant. The ICC [95% confidence interval (CI)] was 0.77 (0.69, 0.85) for TpTe, 0.75 (0.65, 0.85) for QT and 0.60 (0.47, 0.73) for (QT)a. The MDC (ms) was 21, 32 and 26 for TpTe, QT and (QT)a respectively. CONCLUSION: TpTe has excellent repeatability supporting its use as a supplement to QT in observational and clinical studies.

Dispersion Modeling of Traffic-Related Air Pollutant Exposures and Health Effects Among Children with Asthma in Detroit, Michigan.

Vehicular traffic is a major source of ambient air pollution in urban areas. Traffic-related air pollutants, including carbon monoxide, nitrogen oxides, particulate matter less than 2.5 µm in diameter, and diesel exhaust emissions, have been associated with adverse human health effects, especially in areas near major roads. In addition to emissions from vehicles, ambient concentrations of air pollutants include contributions from stationary sources and background (or regional) sources. Although dispersion models have been widely used to evaluate air quality strategies and policies and can represent the spatial and temporal variation in environments near roads, the use of these models in health studies to estimate air pollutant exposures has been relatively limited. This paper summarizes the modeling system used to estimate exposures in the Near-Roadway Exposure and Urban Air Pollutant Study, an epidemiological study that examined 139 children with asthma or symptoms consistent with asthma, most of whom lived near major roads in Detroit, Michigan. Air pollutant concentrations were estimated with a hybrid modeling framework that included detailed inventories of mobile and stationary sources on local and regional scales; the RLINE, AERMOD, and CMAQ dispersion models; and monitored observations of pollutant concentrations. The temporal and spatial variability in emissions and exposures over the 2.5-year study period and at more than 300 home and school locations was characterized. The paper highlights issues with the development and understanding of the significance of traffic-related exposures through the use of dispersion models in urban-scale exposure assessments and epidemiology studies.

Characterization of the Structural Requirements for the NADase Activity of Bacterial Toll/IL-1R domains in a Course-based Undergraduate Research Experience.

TLRs initiate innate immune signaling pathways via Toll/IL-1R (TIR) domains on their cytoplasmic tails. Various bacterial species also express TIR domain-containing proteins that contribute to bacterial evasion of the innate immune system. Bacterial TIR domains, along with the mammalian sterile α and TIR motif-containing protein 1 and TIRs from plants, also have been found to exhibit NADase activity. Initial X-ray crystallographic studies of the bacterial TIR from Acinetobacter baumannii provided insight into bacterial TIR structure but were unsuccessful in cocrystallization with the NAD+ ligand, leading to further questions about the TIR NAD binding site. In this study, we designed a Course-Based Undergraduate Research Experience (CURE) involving 16-20 students per year to identify amino acids crucial for NADase activity of A. baumannii TIR domain protein and the TIR from Escherichia coli (TIR domain-containing protein C). Students used structural data to identify amino acids that they hypothesized would play a role in TIR NADase activity, and created plasmids to express mutated TIRs through site-directed mutagenesis. Mutant TIRs were expressed, purified, and tested for NADase activity. The results from these studies provide evidence for a conformational change upon NAD binding, as was predicted by recent cryogenic electron microscopy and hydrogen-deuterium exchange mass spectrometry studies. Along with corroborating recent characterization of TIR NADases that could contribute to drug development for diseases associated with dysregulated TIR activity, this work also highlights the value of CURE-based projects for inclusion of a diverse group of students in authentic research experiences.

Lipopolysaccharide induction of autophagy is associated with enhanced bactericidal activity in Dictyostelium discoideum.

Innate immune cells respond to microbial invaders using pattern recognition receptors that detect conserved microbial patterns. Among the cellular processes stimulated downstream of pattern recognition machinery is the initiation of autophagy, which plays protective roles against intracellular microbes. We have shown recently that Dictyostelium discoideum, which takes up bacteria for nutritive purposes, may employ pattern recognition machinery to respond to bacterial prey, as D. discoideum cells upregulate bactericidal activity upon stimulation by lipopolysaccharide (LPS). Here we extend these findings, showing that LPS treatment leads to induction of autophagosomal maturation in cells responding to the bacteria Staphylococcus aureus. Cells treated with the autophagy-inducing drug rapamycin clear internalized bacteria at an accelerated rate, while LPS-enhanced clearance of bacteria is reduced in cells deficient for the autophagy-related genes atg1 and atg9. These findings link microbial pattern recognition with autophagy in the social amoeba D. discoideum.

Flipped ophthalmology classroom augmented with case-based learning.

BACKGROUND: Although the flipped classroom model provides an effective way to teach ophthalmology to medical students, there are concerns that it overburdens the learner. The purpose of this study was to assess medical students' perceptions of a case-based flipped classroom style compared with a traditional didactic lecture series and to evaluate the effects of case-based learning on students' confidence in managing common ophthalmic complaints. METHODS: We created an interactive, case-based flipped classroom ophthalmology curriculum. Paired pre- and post-clerkship surveys were distributed to students on the first and last day of the 2-week clerkship. Questions were formatted as statements using a 6-point Likert scale to assess students' prior exposure to a flipped classroom, perceptions of the flipped classroom curriculum, and confidence in evaluating ophthalmic complaints. RESULTS: A total of 75 students were included during the period July 2019 to March 2020. Pre-clerkship questionnaires revealed no preference for either teaching modality. Wilcoxon signed-rank testing comparing pre- and post-clerkship data revealed a significant increase in students' favoring the case-based flipped-classroom model. Participants reported significant reductions in pressure to perform, course burden, and overall anxiety as well as increased confidence in triaging common eye complaints. CONCLUSIONS: The case-based flipped classroom modality prioritizes key learning objectives while increasing student participation and confidence. The reproducibility and accessibility of standardized prepared video lectures and cases may help institutions to better incorporate ophthalmology into preexisting rotations.

Cobbling Together the Myddosome.

An organism's ability to recognize and respond quickly and appropriately to pathogenic stimuli is a fundamental aspect of innate immunity. Harnessing the dynamic nature of fluorescent microscopy and the resolution of cryo-electron microscopy, Moncrieffe et al. (2020) characterize MyD88-only filaments and provide insight into the mechanisms underlying innate immune signaling.

Variability in Observation-based Onroad Emission Constraints from a Near-road Environment.

This study uses Las Vegas near-road measurements of carbon monoxide (CO) and nitrogen oxides (NOx) to test the consistency of onroad emission constraint methodologies. We derive commonly used CO to NOx ratios (ΔCO:ΔNOx) from cross-road gradients and from linear regression using ordinary least squares (OLS) regression and orthogonal regression. The CO to NOx ratios are used to infer NOx emission adjustments for a priori emissions estimates from EPA's MOtor Vehicle Emissions Simulator (MOVES) model assuming unbiased CO. The assumption of unbiased CO emissions may not be appropriate in many circumstances but was implemented in this analysis to illustrate the range of NOx scaling factors that can be inferred based on choice of methods and monitor distance alone. For the nearest road estimates (25m), the cross-road gradient and ordinary least squares (OLS) agree with each other and are not statistically different from the MOVES-based emission estimate while ΔCO:ΔNOx from orthogonal regression is significantly higher than the emitted ratio from MOVES. Using further downwind measurements (i.e., 115m and 300m) increases OLS and orthogonal regression estimates of ΔCO:ΔNOx but not cross-road gradient ΔCO:ΔNOx. The inferred NOx emissions depend on the observation-based method, as well as the distance of the measurements from the roadway and can suggest either that MOVES NOx emissions are unbiased or that they should be adjusted downward by between 10% and 47%. The sensitivity of observation-based ΔCO:ΔNOx estimates to the selected monitor location and to the calculation method characterize the inherent uncertainty of these methods that cannot be derived from traditional standard-error based uncertainty metrics.

Evidence-based anesthesia: fever of unknown origin in parturients and neuraxial anesthesia.

The safety of neuraxial analgesia in febrile patients is controversial. We performed an evidenced-based project in an effort to establish a guideline for our active obstetric clinical practice. Neuraxial anesthesia is generally safe for parturients, and complications are rare; however, serious adverse outcomes can result. Because of the devastating nature of the morbidity, the decision to proceed with a neuraxial anesthetic in the face of infection may be contentious. Fever and sepsis are considered relative contraindications to regional anesthesia; however, epidural anesthesia is a superior method of management of pain during labor. One must also consider that 30% to 40% of patients with chorioamnionitis require cesarean delivery. Because of the increased morbidity and mortality of general anesthesia in this population, it may be reasonable to proceed with regional anesthesia. Based on a review of the literature, it is difficult to estimate the risk of an infrequently occurring event. We recommend evaluation of each individual to determine the risks and benefits of the anesthetic. However, it is prudent to administer antibiotics before the regional anesthetic and adhere to strict aseptic technique. Postprocedure monitoring is essential for early detection and treatment of complications.

Influential factors affecting black carbon trends at four sites of differing distance from a major highway in Las Vegas.

Elevated air pollution levels adjacent to major highways are an ongoing topic of public health concern worldwide. Black carbon (BC), a component of particulate matter (PM) emitted by diesel and gasoline vehicles, was measured continuously via a filter-based light absorption technique over ~ 16 months at four different stations positioned on a perpendicular trajectory to a major highway in Las Vegas, NV. During downwind conditions (winds from the west), BC at 20 m from the highway was 32 and 60% higher than concentrations at 100 and 300 m from the roadway, respectively. Overall highest roadside (20-m site) BC concentrations were observed during the time period of 4 a.m.-8 a.m. under low-speed variable winds (3.02 µg/m3) or downwind conditions (2.84 µg/m3). The 20-m site BC concentrations under downwind conditions are 85% higher on weekday periods compared to weekends during the time period of 4 a.m.-8 a.m. Whereas total traffic volume was higher on weekdays versus weekends and differed by approximately 3% on weekdays versus weekends, similarly, the detected heavy-duty fraction was higher on weekdays versus weekends and differed by approximately 21% on weekdays versus weekend. Low wind speeds predominated during early morning hours, leading to higher BC concentrations during early morning hours despite the maximum traffic volume occurring later in the day. No noticeable impact from the airport or nearby arterial roadways was observed, with the 300-m site remaining the lowest of the four-site network when winds were from the east. Multivariate linear regression analysis revealed that heavy-duty traffic volume, light-duty traffic volume, wind speed, weekday versus weekend, surface friction velocity, ambient temperature, and the background BC concentration were significant predictors of roadside BC concentrations. Comparison of BC and PM2.5 downwind concentration gradients indicates that the BC component contributes substantially to the PM2.5 increase in roadside environments. These results suggest that BC is an important indicator to assess the contribution of primary traffic emissions to near-road PM2.5 concentrations, providing opportunities to evaluate the feasibility and effectiveness of mitigation strategies.

Marking 15 years of the Genetic and Rare Diseases Information Center.

BACKGROUND: The Genetic and Rare Diseases Information Center (GARD), a program of the National Center for Advancing Translational Sciences, was established in 2002 to assist the public in finding reliable, timely, and easy-to-understand information about genetic and/or rare diseases in English or Spanish. OBJECTIVE: A review of longitudinal data on GARD inquiries from 2002 to 2016 and assessment of the results of two user satisfaction surveys were conducted to understand the demographics and needs of GARD customers over time. METHODS: Since 2002, GARD has collected anonymized data while responding to questions received via e-mail, website, telephone, fax, letter, or TTY. Between 2002 and 2016 GARD received a total of 60,106 inquiries. User satisfaction surveys were conducted in 2006 and 2014, in which users self-selected to participate. RESULTS: The annual number of inquiries has risen steadily since 2002. Inquiries are overwhelmingly from educated female patients, family, and friends seeking disease-specific information, treatment options, referrals, and research studies. Most users report satisfaction with the experience. CONCLUSIONS: Rare disease patients and their families face challenges in finding information about their symptoms or diagnosis, prognosis, treatment options, significance for family members, and research opportunities. Lack of available clinical expertise can leave patients, their family, and friends with little choice but to become knowledgeable on their own. GARD fills a critical need by providing the public with vetted, evidence-based information that empowers people to engage in their own health care and seek research studies of relevance.

Estimation of on-road NO2 concentrations, NO2/NOX ratios, and related roadway gradients from near-road monitoring data.

This paper describes a new regression modeling approach to estimate on-road nitrogen dioxide (NO2) and oxides of nitrogen (NOX) concentrations and near-road spatial gradients using data from a near-road monitoring network. Field data were collected in Las Vegas, NV at three monitors sited 20, 100, and 300 m from Interstate-15 between December, 2008 and January, 2010. Measurements of NO2 and NOX were integrated over 1-hour intervals and matched with meteorological data. Several mathematical transformations were tested for regressing pollutant concentrations against distance from the roadway. A logit-ln model was found to have the best fit (R2 = 94.7%) and also provided a physically realistic profile. The mathematical model used data from the near-road monitors to estimate on-road concentrations and the near-road gradient over which mobile source pollutants have concentrations elevated above background levels. Average and maximum on-road NO2 concentration estimates were 33 ppb and 105 ppb, respectively. Concentration gradients were steeper in the morning and late afternoon compared with overnight when stable conditions preclude mixing. Estimated on-road concentrations were also highest in the late afternoon. Median estimated on-road and gradient NO2 concentrations were lower during summer compared with winter, with a steeper gradient during the summer, when convective mixing occurs during a longer portion of the day On-road concentration estimates were higher for winds perpendicular to the road compared with parallel winds and for atmospheric stability with neutral-to-unstable atmospheric conditions. The concentration gradient with increasing distance from the road was estimated to be sharper for neutral-to-unstable conditions when compared with stable conditions and for parallel wind conditions compared with perpendicular winds. A regression of the NO2/NOX ratios yielded on-road ratios ranging from 0.25 to 0.35, substantially higher than the anticipated tail-pipe emissions ratios. The results from the ratios also showed that the diurnal cycle of the background NO2/NOX ratios were a driving factor in the on-road and downwind NO2/NOX ratios.