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Abstract Riboflavin transporter deficiency (RTD) is a genetic disorder of reduced riboflavin (vitamin B2) uptake that causes progressive, multifocal neurological dysfunction. Most patients present in early childhood; if patients present later in life, symptoms usually develop more gradually. We report three previously healthy young adults, who developed rapidly progressive neurological symptoms after decreasing dietary intake of meat and dairy. After a diagnostic odyssey, the diagnosis of a riboflavin transporter deficiency was made. Treatment with high dose oral riboflavin (20–40 mg/kg/day) partially reversed symptoms. This case series highlights that reduced riboflavin intake as a result of dietary changes can unmask RTD at a later age. We emphasize the importance of early recognition of this progressive and potentially lethal disease and show that timely treatment with high dose riboflavin is highly effective.

Abstract Immune dysregulation is a hallmark of chronic lymphocytic leukemia (CLL). Anti‐CD20 antibodies (e.g., rituximab [R]) can be combined with venetoclax (Ven) to treat CLL. However, anti‐CD20 antibodies can increase hypogammaglobulinemia risk, while the effects of Ven on immune dysregulation are still uncertain. We report long‐term immune changes in VenR‐ and bendamustine‐R (BR)‐treated patients with relapsed/refractory CLL in the MURANO trial (NCT02005471). Patients were randomized to fixed‐duration VenR (2 years Ven; VenR for the first 6 months) or BR (6 months). Immune cell levels were evaluated at the end of combination treatment (EOCT), end of treatment (EOT; VenR arm only), and 12 and 24 months post‐EOCT. Overall, 130/194 VenR‐ and 134/195 BR‐treated patients completed treatment without progressive disease. In patients who completed VenR combination therapy, median immunoglobulin (Ig)G and IgM levels decreased from baseline to EOT (p ≤ 0.01 and p ≤ 0.0001, respectively); by 24 months, post‐EOT IgG had returned to baseline level and IgM had increased from baseline (p ≤ 0.001). Median IgA levels increased from baseline to 12 (p ≤ 0.0001) and 24 months post‐EOT (p ≤ 0.0001). In BR‐treated patients, changes in IgG, IgA, and IgM levels across the assessed time points were not significant, and by 24 months, post‐EOCT IgG, IgA, and IgM were above baseline levels. Grade ≥3 infection rates on treatment were low. Overall, immune recovery was observed with VenR and BR, with stabilization of Ig levels after treatment. Post‐treatment infection rates were generally low, making these very tolerable therapies for CLL.

Background We examined the association of multilevel social determinants of health with incident apparent treatment‐resistant hypertension (aTRH). Methods and Results We analyzed data from 2774 White and 2257 Black US adults from the REGARDS (Reasons for Geographic and Racial Differences in Stroke) study taking antihypertensive medication without aTRH at baseline to estimate the association of social determinants of health with incident aTRH. Selection of social determinants of health was guided by the Healthy People 2030 domains of education, economic stability, social context, neighborhood environment, and health care access. Blood pressure (BP) was measured during study visits, and antihypertensive medication classes were identified through a pill bottle review. Incident aTRH was defined as (1) systolic BP ≥140 mm Hg or diastolic BP ≥90 mm Hg, or systolic BP ≥130 mm Hg or diastolic BP ≥80 mm Hg for those with diabetes or chronic kidney disease while taking ≥3 classes of antihypertensive medication or (2) taking ≥4 classes of antihypertensive medication regardless of BP level, at the follow‐up visit. Over a median 9.5 years of follow‐up, 15.9% of White and 24.0% of Black adults developed aTRH. A percent of the excess aTRH risk among Black versus White adults was mediated by low education (14.2%), low income (16.0%), not seeing a friend or relative in the past month (8.1%), not having someone to care for them if ill or disabled (7.6%), lack of health insurance (10.6%), living in a disadvantaged neighborhood (18.0%), and living in states with poor public health infrastructure (6.0%). Conclusions Part of the association between race and incident aTRH risk was mediated by social determinants of health.

Background Higher cardiovascular health (CVH) score is associated with lower risks of cardiovascular disease (CVD) and mortality in the general population. However, it is unclear whether cumulative CVH is associated with CVD, end‐stage kidney disease (ESKD), and death in patients with chronic kidney disease. Methods and Results Among individuals from the prospective CRIC (Chronic Renal Insufficiency Cohort) Study, we used the percentage of the maximum possible CVH score attained from baseline to the year 5 visit to calculate cumulative CVH score. Multivariable‐adjusted Cox proportional hazards regression was used to investigate the associations of cumulative CVH with risks of adjudicated CVD (myocardial infarction, stroke, and heart failure), ESKD, and all‐cause mortality. A total of 3939 participants (mean age, 57.7 years; 54.9% men) were included. The mean (SD) cumulative CVH score attained during 5 years was 55.5% (12.3%). Over a subsequent median 10.2‐year follow‐up, 597 participants developed CVD, 656 had ESKD, and 1324 died. A higher cumulative CVH score was significantly associated with lower risks of CVD, ESKD, and mortality, independent of the CVH score at year 5. Multivariable‐adjusted hazard ratios and 95% CIs per 10% higher cumulative CVH score during 5 years were 0.81 (0.69–0.95) for CVD, 0.82 (0.70–0.97) for ESKD, and 0.80 (0.72–0.89) for mortality. Conclusions Among patients with chronic kidney disease stages 2 to 4, a better CVH status maintained throughout 5 years is associated with lower risks of CVD, ESKD, and all‐cause mortality. The findings support the need for interventions to maintain ideal CVH status for prevention of adverse outcomes in the population with chronic kidney disease.

Amino acid‐based surfactants are valuable compounds for cosmetic formulations. The chemical synthesis of acyl amino acids is conventionally performed by the Schotten–Baumann reaction using fatty acyl chlorides, but aminoacylases have also been investigated for use in biocatalytic synthesis with free fatty acids. Aminoacylases and their properties are diverse; they belong to different peptidase families and show differences in substrate specificity and biocatalytic potential. Bacterial aminoacylases capable of synthesis have been isolated from Burkholderia, Mycolicibacterium, and Streptomyces. Although several proteases and peptidases from S. griseus have been described, no aminoacylases from this species have been identified yet. In this study, we investigated two novel enzymes produced by S. griseus DSM 40236T. We identified and cloned the respective genes and recombinantly expressed an α‐aminoacylase (EC3.5.1.14), designated SgAA, and an ε‐lysine acylase (EC3.5.1.17), designated SgELA, in S. lividans TK23. The purified aminoacylase SgAA was biochemically characterized, focusing on its hydrolytic activity to determine temperature‐ and pH optima and stabilities. The aminoacylase could hydrolyze various acetyl amino acids at the Nα‐position with a broad specificity regarding the sidechain. Substrates with longer acyl chains, like lauroyl amino acids, were hydrolyzed to a lesser extent. Purified aminoacylase SgELA specific for the hydrolysis of Nε‐acetyl‐l‐lysine was unstable and lost its enzymatic activity upon storage for a longer period but could initially be characterized. The pH optimum of SgELA was pH 8.0. While synthesis of acyl amino acids was not observed with SgELA, SgAA catalyzed the synthesis of lauroyl‐methionine.

Abstract Understanding dispersal potential, or the probability a species will move a given distance, under different environmental conditions is essential to predicting species' ability to move across the landscape and track shifting ecological niches. Two important drivers of dispersal ability are climatic differences and variations in local habitat type. Despite the likelihood these global drivers act simultaneously on plant populations, and thus dispersal potential is likely to change as a result, their combined effects on dispersal are rarely examined. To understand the effect of climate and varying habitat types on dispersal potential, we studied Geum triflorum—a perennial grassland species that spans a wide range of environments, including both prairie and alvar habitats. We explored how the climate of the growing season and habitat type (prairie vs. alvar) interact to alter dispersal potential. We found a consistent interactive effect of climate and habitat type on dispersal potential. Across prairie populations, an increased number of growing degree days favored traits that increase dispersal potential or the probability of dispersing farther distances. However, for alvar populations, dispersal potential tended to decrease as the number of growing degree days increased. Our findings suggest that under continued warming, populations in prairie habitats will benefit from increased gene flow, while alvar populations will become increasingly segregated, with reduced potential to track shifting fitness optima.

Abstract Impranil® DLN‐SD is a poly(ester‐urethane) (PEU) that is widely used as coating material for textiles to fine‐tune and improve their properties. Since coatings increase the complexity of such plastic materials, they can pose a hindrance for sustainable end‐of‐life solutions of plastics using enzymes or microorganisms. In this study, we isolated Halopseudomonas formosensis FZJ due to its ability to grow on Impranil DLN‐SD and other PEUs as sole carbon sources. The isolated strain was exceptionally thermotolerant as it could degrade Impranil DLN‐SD at up to 50°C. We identified several putative extracellular hydrolases of which the polyester hydrolase Hfor_PE‐H showed substrate degradation of Impranil DLN‐SD and thus was purified and characterized in detail. Hfor_PE‐H showed moderate temperature stability (Tm = 53.9°C) and exhibited activity towards Impranil DLN‐SD as well as polyethylene terephthalate. Moreover, we revealed the enzymatic release of monomers from Impranil DLN‐SD by Hfor_PE‐H using GC‐ToF‐MS and could decipher the associated metabolic pathways in H. formosensis FZJ. Overall, this study provides detailed insights into the microbial and enzymatic degradation of PEU coatings, thereby deepening our understanding of microbial coating degradation in both contained and natural environments. Moreover, the study highlights the relevance of the genus Halopseudomonas and especially the novel isolate and its enzymes for future bio‐upcycling processes of coated plastic materials.

Abstract Pseudomonas species have become promising cell factories for the production of natural products due to their inherent robustness. Although these bacteria have naturally evolved strategies to cope with different kinds of stress, many biotechnological applications benefit from engineering of optimised chassis strains with specially adapted tolerance traits. Here, we explored the formation of outer membrane vesicles (OMV) of Pseudomonas putida KT2440. We found OMV production to correlate with the recombinant production of a natural compound with versatile beneficial properties, the tripyrrole prodigiosin. Further, several P. putida genes were identified, whose up‐ or down‐regulated expression allowed controlling OMV formation. Finally, genetically triggering vesiculation in production strains of the different alkaloids prodigiosin, violacein, and phenazine‐1‐carboxylic acid, as well as the carotenoid zeaxanthin, resulted in up to three‐fold increased product yields. Consequently, our findings suggest that the construction of robust strains by genetic manipulation of OMV formation might be developed into a useful tool which may contribute to improving limited biotechnological applications.

Abstract The Halopseudomonas species, formerly classified as Pseudomonas pertucinogena lineage, form a unique phylogenetic branch within the Pseudomonads. Most strains have recently been isolated from challenging habitats including oil‐ or metal‐polluted sites, deep sea, and intertidal zones, suggesting innate resilience to physical and chemical stresses. Despite their comparably small genomes, these bacteria synthesise several biomolecules with biotechnological potential and a role in the degradation of anthropogenic pollutants has been suggested for some Halopseudomonads. Until now, these bacteria are not readily amenable to existing cultivation and cloning methods. We addressed these limitations by selecting four Halopseudomonas strains of particular interest, namely H. aestusnigri, H. bauzanensis, H. litoralis, and H. oceani to establish microbiological and molecular genetic methods. We found that C4‐C10 dicarboxylic acids serve as viable carbon sources in both complex and mineral salt cultivation media. We also developed plasmid DNA transfer protocols and assessed vectors with different origins of replication and promoters inducible with isopropyl‐β‐d‐thiogalactopyranoside, l‐arabinose, and salicylate. Furthermore, we have demonstrated the simultaneous genomic integration of expression cassettes into one and two attTn7 integration sites. Our results provide a valuable toolbox for constructing robust chassis strains and highlight the biotechnological potential of Halopseudomonas strains.

Abstract Introduction Adolescence and young adulthood are especially critical times to learn about mental health, given that 75% of mental health issues are developed by the age of 24. Animations have great potential to effectively deliver mental health information to young people. A series of five short animated films to promote mental health literacy were created with and for young people in partnership with the multi‐award‐winning independent animation studio, Aardman Animations. The aim of this study was to explore young people's perceptions of the cocreated animated films. Methods Seven Youth Juries were conducted to capture young people's opinions and recommendations about the content related to mental health literacy and presentation style of the cocreated animated films. Thematic analysis was used to analyse the audio transcripts. Results Many participants reported a view that the animated films had the potential to promote mental health literacy, especially for understanding mental health and reducing stigma. Some recommendations were provided to improve the films, such as including subtitles and having a better transition to the companion website. Conclusion Cocreated animations have great potential to promote the mental health literacy of young people. We hope that the findings from the present study will inform future media development to make them as effective as possible. Patient or Public Contribution Young people were actively involved in the development, production, implementation and evaluation (up to the time before data analysis) of the animated films.

Abstract Ponds host a variety of invertebrate species and contribute greatly to global biodiversity. Aquaculture influences macroinvertebrate diversity and productivity in ponds through several practices, such as macrophyte and water management. Fish stocking is also considered controversial for preserving biodiversity through the direct predation upon natural species and changes induced on the biotope. An experiment examined whether compartmentalized ponds with temporarily fish‐free areas had higher fish productivity and macroinvertebrate abundance and diversity than open ponds. The experimental design consisted of two treatments—compartmentalized (C) or open (O)—each applied to three ponds. Roach (Rutilus rutilus), tench (Tinca tinca) and common carp (Cyprinus carpio) were stocked in the ponds in March 2021. Juvenile pikeperch (Sander lucioperca) were stocked in the ponds in June. In the C ponds, three areas were created and opened successively: (C1) corresponding to ¼ of the pond surface to host roach, tench and common carp from March to May; (C2) ¼ of the pond surface restricted to fish from March to May; and (C3) ½ of the surface restricted to fish from March to July, except for juveniles of pikeperch which were stocked in June. We investigated patterns in abundance, dry biomass and productivity of macroinvertebrates four times from March to October. This article presents observed macroinvertebrate abundances and weighted dry biomass, and productivity estimated from them. Overall, 77,749 individuals were identified, of which one‐third were Chironomini and another one‐third were Oligochaeta. The invasive red swamp crayfish (Procambarus clarkii) was found in one pond in October. The two highest taxonomic richness values were found in C ponds (71 and 69 taxa). The lowest taxonomic richness (61 taxa) was in an O pond. Although dry biomass was clearly higher in the C ponds in March, no tendency could be seen between C and O ponds throughout the experiment. No difference in productivity was found between the C and O ponds among the experiment. By reporting macroinvertebrate abundance, biomass, productivity, size classes, developmental stages and high‐resolution taxonomic identification in a freshwater polyculture system, this dataset is one of the first of its kind.

Objective To understand perspectives of mental health care providers regarding barriers and drivers of adopting a medication ingestible event monitoring (IEM) system in clinical practice. Methods Between April and October 2019, a cross‐sectional, online survey was conducted among 131 prescribing clinicians and 119 non‐prescribing clinicians providing care to patients with major depressive disorder, bipolar disorder, and schizophrenia. Results Most prescribing clinicians were physicians (79.4%) while most non‐prescribing clinicians (52.9%) were licensed clinical social workers, followed by counselors (30.8%), clinical psychologists (13.4%), and case managers (2.5%). Most respondents (93.2%) reported that clinicians can influence adherence, that the IEM technology was in their patients' best interest (63.6%), and a willingness to beta test the technology (54.8%). Support was positively associated with prescribing clinicians (OR: 2.2; 95% CI: 1.1, 4.5), belief that antipsychotics reduce the health, social, or financial consequences of the condition (OR: 3.8; 95% CI: 1.3, 11.0), concern for patients' well‐being without monitoring (OR: 3.3; 95% CI: 1.2, 8.7), and belief the technology will enhance clinical alliance (OR: 3.1; 95% CI: 1.5, 6.3) or improve patient engagement (OR: 3.0; 95% CI: 1.5, 6.2). Support was inversely related to concerns about appropriate follow‐up actions (OR: 0.4; 95% CI: 0.2, 0.9) and responsibilities (OR: 0.3; 95% CI: 0.1, 0.8) when using the technology. Conclusions Our results suggest that IEM sensor technology adoption will depend upon additional evidence that patients will actively engage in the use of the technology, will benefit from the technology through improved outcomes, and that the additional burden placed upon providers is minimal compared to the potential benefit.

Abstract Beaver dam mimicry is an emergent conservation practice. We evaluated the influence of constructed riffles, a unique type of beaver mimicry aimed to store water and allow fish passage, on habitat for fishes in one control reach and one manipulated reach with mimicry structures added. The beaver mimicry reach had deeper pool habitats and deeper and wider riffle habitats compared to an unmanipulated control reach. Dissolved oxygen was similar among reaches, averaging 8.7 ± 0.2 and 8.9 mg/L in the beaver mimicry and control reaches, respectively. Sediment size was also similar among reaches, with a D50 of 8.1 and 10.6 mm in the beaver mimicry and control reaches, respectively. The beaver mimicry reach had little to no overhanging bank vegetation or riparian vegetation shade cover, while the control had 38% of its bank covered by canopy and 56% overhung by vegetation. These riparian characteristics result from a legacy of livestock grazing and lack of consistent vegetation planting during restoration. Longnose dace (Rhinichthys cataractae) and white sucker (Catostomus commersonii) dominated in the beaver mimicry reach, together comprising 70% of the fish assemblage post‐structure installation. Arctic grayling (Thymallus arcticus) was not found in the beaver mimicry reach but was present in the control, albeit in small numbers of only 3% of the assemblage post‐structure installation. These results highlight the need to consider both in‐stream and riparian habitat features for fishes, as well as timescales of both hydrological and ecological outcomes in restoration design.

Abstract The authors examined the proportion of US adults that would have their high blood pressure (BP) status changed if systolic BP (SBP) and diastolic BP (DBP) were measured with systematic bias and/or random error versus following a standardized protocol. Data from the 2017–2018 National Health and Nutrition Examination Survey (NHANES; n = 5176) were analyzed. BP was measured up to three times using a mercury sphygmomanometer by a trained physician following a standardized protocol and averaged. High BP was defined as SBP ≥130 mm Hg or DBP ≥80 mm Hg. Among US adults not taking antihypertensive medication, 32.0% (95%CI: 29.6%,34.4%) had high BP. If SBP and DBP were measured with systematic bias, 5 mm Hg for SBP and 3.5 mm Hg for DBP higher and lower than in NHANES, the proportion with high BP was estimated to be 44.4% (95%CI: 42.6%,46.2%) and 21.9% (95%CI 19.5%,24.4%). Among US adults taking antihypertensive medication, 60.6% (95%CI: 57.2%,63.9%) had high BP. If SBP and DBP were measured 5 and 3.5 mm Hg higher and lower than in NHANES, the proportion with high BP was estimated to be 71.8% (95%CI: 68.3%,75.0%) and 48.4% (95%CI: 44.6%,52.2%), respectively. If BP was measured with random error, with standard deviations of 15 mm Hg for SBP and 7 mm Hg for DBP, 21.4% (95%CI: 19.8%,23.0%) of US adults not taking antihypertensive medication and 20.5% (95%CI: 17.7%,23.3%) taking antihypertensive medication had their high BP status re‐categorized. In conclusions, measuring BP with systematic or random errors may result in the misclassification of high BP for a substantial proportion of US adults.

Abstract Objective To investigate local short‐term neuroplasticity elicited by subthalamic, thalamic, and pallidal deep brain stimulation (DBS) for movement disorders. Methods During DBS surgery, we delivered pairs of stimulus pulses with both circular and directional leads across 90 interstimulus intervals in 17 participants and recorded local field potentials from unused contacts on the implanted electrode array. We removed the stimulus artifact, validated the neural origin of the underlying signals, and examined short‐term plasticity as a function of interstimulus interval and DBS target, using linear mixed effects models. Results DBS evokes short latency local field potentials that are readily detected with both circular and directional leads at all stimulation targets (0.31 ± 0.10 msec peak latency, mean ± SD). Peak amplitude, area, and latency are modified strongly by interstimulus interval (P

Abstract Investigations of the genetic structure of populations over the entire range of a species yield valuable information about connectivity among populations. Seabirds are an intriguing taxon in this regard because they move extensively when not breeding, facilitating intermixing of populations, but breed consistently on the same isolated islands, restricting gene flow among populations. The degree of genetic structuring of populations varies extensively among seabird species but they have been understudied in their tropical ranges. Here, we address this across a broad spatial scale by using microsatellite and mitochondrial data to explore the population connectivity of 13 breeding populations representing the six subspecies of the white‐tailed tropicbird (Phaethon lepturus) in the Atlantic, Indian, and Pacific Oceans. Our primary aim was to identify appropriate conservation units for this little known species. Three morphometric characters were also examined in the subspecies. We found a clear pattern of population structuring with four genetic groups. The most ancient and the most isolated group was in the northwestern Atlantic Ocean. The South Atlantic populations and South Mozambique Channel population on Europa were genetically isolated and may have had a common ancestor. Birds from the Indo‐Pacific region showed unclear and weak genetic differentiation. This structuring was most well defined from nuclear and mtDNA markers but was less well resolved by morphological data. The validity of classifying white‐tailed tropicbirds into six distinct subspecies is discussed in light of our new findings. From a conservation standpoint our results highlight that the three most threatened conservation units for this species are the two subspecies of the tropical North and South Atlantic Oceans and that of Europa Island in the Indian Ocean.

Summary Marine habitats represent a prolific source for molecules of biotechnological interest. In particular, marine bacteria have attracted attention and were successfully exploited for industrial applications. Recently, a group of Pseudomonas species isolated from extreme habitats or living in association with algae or sponges were clustered in the newly established Pseudomonas pertucinogena lineage. Remarkably for the predominantly terrestrial genus Pseudomonas, more than half (9) of currently 16 species within this lineage were isolated from marine or saline habitats. Unlike other Pseudomonas species, they seem to have in common a highly specialized metabolism. Furthermore, the marine members apparently possess the capacity to produce biomolecules of biotechnological interest (e.g. dehalogenases, polyester hydrolases, transaminases). Here, we summarize the knowledge regarding the enzymatic endowment of the marine Pseudomonas pertucinogena bacteria and report on a genomic analysis focusing on the presence of genes encoding esterases, dehalogenases, transaminases and secondary metabolites including carbon storage compounds.

Summary Hydrolases acting on polyesters like cutin, polycaprolactone or polyethylene terephthalate (PET) are of interest for several biotechnological applications like waste treatment, biocatalysis and sustainable polymer modifications. Recent studies suggest that a large variety of such enzymes are still to be identified and explored in a variety of microorganisms, including bacteria of the genus Pseudomonas. For activity‐based screening, methods have been established using agar plates which contain nanoparticles of polycaprolactone or PET prepared by solvent precipitation and evaporation. In this protocol article, we describe a straightforward agar plate‐based method using emulsifiable artificial polyesters as substrates, namely Impranil® DLN and liquid polycaprolactone diol (PLD). Thereby, the currently quite narrow set of screening substrates is expanded. We also suggest optional pre‐screening with short‐chain and middle‐chain‐length triglycerides as substrates to identify enzymes with lipolytic activity to be further tested for polyesterase activity. We applied these assays to experimentally demonstrate polyesterase activity in bacteria from the P. pertucinogena lineage originating from contaminated soils and diverse marine habitats.

Summary Bacterial gene clusters, which represent a genetic treasure trove for secondary metabolite pathways, often need to be activated in a heterologous host to access the valuable biosynthetic products. We provide here a detailed protocol for the application of the yTREX ‘gene cluster transplantation tool’: Via yeast recombinational cloning, a gene cluster of interest can be cloned in the yTREX vector, which enables the robust conjugational transfer of the gene cluster to bacteria like Pseudomonas putida, and their subsequent transposon Tn5‐based insertion into the host chromosome. Depending on the gene cluster architecture and chromosomal insertion site, the respective pathway genes can be transcribed effectively from a chromosomal promoter, thereby enabling the biosynthesis of a natural product. We describe workflows for the design of a gene cluster expression cassette, cloning of the cassette in the yTREX vector by yeast recombineering, and subsequent transfer and expression in P. putida. As an example for yTREX‐based transplantation of a natural product biosynthesis, we provide details on the cloning and activation of the phenazine‐1‐carboxylic acid biosynthetic genes from Pseudomonas aeruginosa in P. putidaKT2440 as well as the use of β‐galactosidase‐encoding lacZ as a reporter of production levels.

Abstract Introduction Practical algorithms predicting the probability of amyloid pathology among patients with subjective cognitive decline or mild cognitive impairment may help clinical decisions regarding confirmatory biomarker testing for Alzheimer's disease. Methods Algorithm feature selection was conducted with Alzheimer's Disease Neuroimaging Initiative and Australian Imaging, Biomarkers and Lifestyle Flagship Study of Ageing data. Probability algorithms were developed in Alzheimer's Disease Neuroimaging Initiative using nested cross‐validation accompanied by stratified subsampling to obtain 1000 internally validated decision trees. Semi‐independent validation was conducted using Australian Imaging, Biomarkers and Lifestyle Flagship Study of Ageing. Independent external validation was conducted in the population‐based Mayo Clinic Study of Aging. Results Two algorithms were developed using age and normalized immediate recall z‐scores, with or without apolipoprotein E ε4 carrier status. Both algorithms had robust performance across data sets and when substituting different recall memory tests. Discussion The statistical framework resulted in robust probability estimation. Application of these algorithms may assist in clinical decision‐making for further testing to diagnose amyloid pathology.

Abstract The global road network is expanding at an unprecedented rate, threatening the persistence of many species. Yet, even for the most endangered wildlife, crucial information on the distance up to which roads impact species abundance is lacking. Here we use ecological threshold analysis to quantify the road‐effect zone (REZ) for the critically endangered western chimpanzee (Pan troglodytes verus). We found: (1) the REZ extends 5.4 km (95% CI [4.9–5.8 km]) from minor roads and 17.2 km (95% CI [15.8–18.6]) from major roads, the latter being more than three times wider than a previous estimate of the average REZ for mammals; and (2) only 4.3% of the chimpanzees’ range is not impacted by existing roads. These findings reveal the high sensitivity and susceptibility of nonhuman primates to roads across West Africa, a region undergoing rapid development, and can inform the implementation of more effective guidelines to mitigate road impacts.

Background It is unclear whether the recent increase in the number of heart transplants performed annually in the United States is only because of higher availability of donors and if it affected recipients’ survival. Methods and Results We examined characteristics of donors and recipients from 2008 to 2012 (n=11 654) and 2013 to 2017 (n=14 556) and compared them with 2003 to 2007 (n=10 869). Cox models examined 30‐day and 1‐year risk of recipients’ death post transplant. From 2013 to 2017, there was an increase in the number of transplanted hearts and number of donor offers but an overall decline in the ratio of hearts transplanted to available donors. Donors between 2013 and 2017 were older, heavier, more hypertensive, diabetic, and likely to have abused illicit drugs compared with previous years. Drug overdose and hepatitis C positive donors were additional contributors to donor risk in recent years. In Cox models, risk of death post transplant between 2013 and 2017 was 15% lower at 30 days (hazard ratio [HR] 0.85; 95% CI, 0.74–0.98) and 21% lower at 1 year (HR, 0.79; 95% CI, 0.73–0.87) and between 2008 and 2012 was 9% lower at 30 days (HR, 0.91; 95% CI, 0.79–1.05) and 14% lower at 1 year (HR, 0.86; 95% CI, 0.79–0.94) compared with 2003 to 2007. Conclusions Despite a substantial increase in heart donor offers in recent years, the ratio of transplants performed to available donors has decreased. Even though hearts from donors who are older, more hypertensive, and have diabetes mellitus are being used, overall recipient survival continues to improve. Broader acceptance of drug overdose and hepatitis C positive donors may increase the number and percentage of heart transplants further without jeopardizing short‐term outcomes.