Advances in euglenoid genomics: unravelling the fascinating biology of a complex clade.

Euglenids have long been studied due to their unique physiology and versatile metabolism, providing underpinnings for much of our understanding of photosynthesis and biochemistry, and a growing opportunity in biotechnology. Until recently there has been a lack of genetic studies due to their large and complex genomes, but recently new technologies have begun to unveil their genetic capabilities. Whilst much research has focused on the model organism Euglena gracilis, other members of the euglenids have now started to receive due attention. Currently only poor nuclear genome assemblies of E. gracilis and Rhabdomonas costata are available, but there are many more plastid genome sequences and an increasing number of transcriptomes. As more assemblies become available, there are great opportunities to understand the fundamental biology of these organisms and to exploit them for biotechnology.

Eukaryotic cellular intricacies shape mitochondrial proteomic complexity.

Mitochondria have been fundamental to the eco-physiological success of eukaryotes since the last eukaryotic common ancestor (LECA). They contribute essential functions to eukaryotic cells, above and beyond classical respiration. Mitochondria interact with, and complement, metabolic pathways occurring in other organelles, notably diversifying the chloroplast metabolism of photosynthetic organisms. Here, we integrate existing literature to investigate how mitochondrial metabolism varies across the landscape of eukaryotic evolution. We illustrate the mitochondrial remodelling and proteomic changes undergone in conjunction with major evolutionary transitions. We explore how the mitochondrial complexity of the LECA has been remodelled in specific groups to support subsequent evolutionary transitions, such as the acquisition of chloroplasts in photosynthetic species and the emergence of multicellularity. We highlight the versatile and crucial roles played by mitochondria during eukaryotic evolution, extending from its huge contribution to the development of the LECA itself to the dynamic evolution of individual eukaryote groups, reflecting both their current ecologies and evolutionary histories.

Association of Habitual Physical Activity With Cardiovascular Disease Risk.

RATIONALE: A sedentary lifestyle is associated with increased risk for cardiovascular disease (CVD). Smartwatches enable accurate daily activity monitoring for physical activity measurement and intervention. Few studies, however, have examined physical activity measures from smartwatches in relation to traditional risk factors associated with future risk for CVD. OBJECTIVE: To investigate the association of habitual physical activity measured by smartwatch with predicted CVD risk in adults. METHODS AND RESULTS: We enrolled consenting FHS (Framingham Heart Study) participants in an ongoing eFHS (electronic Framingham Heart Study) at the time of their FHS research center examination. We provided participants with a smartwatch (Apple Watch Series 0) and instructed them to wear it daily, which measured their habitual physical activity as the average daily step count. We estimated the 10-year predicted risk of CVD using the American College of Cardiology/American Heart Association 2013 pooled cohort risk equation. We estimated the association between physical activity and predicted risk of CVD using linear mixed effects models adjusting for age, sex, wear time, and familial structure. Our study included 903 eFHS participants (mean age 53±9 years, 61% women, 9% non-White) who wore the smartwatch ≥5 hours per day for ≥30 days. Median daily step count was similar among men (7202 with interquartile range 3619) and women (7260 with interquartile range 3068; P=0.52). Average 10-year predicted CVD risk was 4.5% (interquartile range, 6.1%) for men and 1.2% (interquartile range, 2.2%) for women (P=1.3×10-26). Every 1000 steps higher habitual physical activity was associated with 0.18% lower predicted CVD risk (P=3.2×10-4). The association was attenuated but remained significant after further adjustment for body mass index (P=0.01). CONCLUSIONS: In this community-based sample of adults, higher daily physical activity measured by a study smartwatch was associated with lower predicted risk of CVD. Future research should examine the longitudinal association of prospectively measured daily activity and incident CVD.

A Uniquely Complex Mitochondrial Proteome from Euglena gracilis.

Euglena gracilis is a metabolically flexible, photosynthetic, and adaptable free-living protist of considerable environmental importance and biotechnological value. By label-free liquid chromatography tandem mass spectrometry, a total of 1,786 proteins were identified from the E. gracilis purified mitochondria, representing one of the largest mitochondrial proteomes so far described. Despite this apparent complexity, protein machinery responsible for the extensive RNA editing, splicing, and processing in the sister clades diplonemids and kinetoplastids is absent. This strongly suggests that the complex mechanisms of mitochondrial gene expression in diplonemids and kinetoplastids occurred late in euglenozoan evolution, arising independently. By contrast, the alternative oxidase pathway and numerous ribosomal subunits presumed to be specific for parasitic trypanosomes are present in E. gracilis. We investigated the evolution of unexplored protein families, including import complexes, cristae formation proteins, and translation termination factors, as well as canonical and unique metabolic pathways. We additionally compare this mitoproteome with the transcriptome of Eutreptiella gymnastica, illuminating conserved features of Euglenida mitochondria as well as those exclusive to E. gracilis. This is the first mitochondrial proteome of a free-living protist from the Excavata and one of few available for protists as a whole. This study alters our views of the evolution of the mitochondrion and indicates early emergence of complexity within euglenozoan mitochondria, independent of parasitism.

The distinctive flagellar proteome of Euglena gracilis illuminates the complexities of protistan flagella adaptation.

The eukaryotic flagellum/cilium is a prominent organelle with conserved structure and diverse functions. Euglena gracilis, a photosynthetic and highly adaptable protist, employs its flagella for both locomotion and environmental sensing. Using proteomics of isolated E. gracilis flagella we identify nearly 1700 protein groups, which challenges previous estimates of the protein complexity of motile eukaryotic flagella. We not only identified several unexpected similarities shared with mammalian flagella, including an entire glycolytic pathway and proteasome, but also document a vast array of flagella-based signal transduction components that coordinate gravitaxis and phototactic motility. By contrast, the pellicle was found to consist of > 900 protein groups, containing additional structural and signalling components. Our data identify significant adaptations within the E. gracilis flagellum, many of which are clearly linked to the highly flexible lifestyle.

Association of Habitual Physical Activity With Home Blood Pressure in the Electronic Framingham Heart Study (eFHS): Cross-sectional Study.

BACKGROUND: When studied in community-based samples, the association of physical activity with blood pressure (BP) remains controversial and is perhaps dependent on the intensity of physical activity. Prior studies have not explored the association of smartwatch-measured physical activity with home BP. OBJECTIVE: We aimed to study the association of habitual physical activity with home BP. METHODS: Consenting electronic Framingham Heart Study (eFHS) participants were provided with a study smartwatch (Apple Watch Series 0) and Bluetooth-enabled home BP cuff. Participants were instructed to wear the watch daily and transmit BP values weekly. We measured habitual physical activity as the average daily step count determined by the smartwatch. We estimated the cross-sectional association between physical activity and average home BP using linear mixed effects models adjusting for age, sex, wear time, antihypertensive drug use, and familial structure. RESULTS: We studied 660 eFHS participants (mean age 53 years, SD 9 years; 387 [58.6%] women; 602 [91.2%] White) who wore the smartwatch 5 or more hours per day for 30 or more days and transmitted three or more BP readings. The mean daily step count was 7595 (SD 2718). The mean home systolic and diastolic BP (mmHg) were 122 (SD 12) and 76 (SD 8). Every 1000 increase in the step count was associated with a 0.49 mmHg lower home systolic BP (P=.004) and 0.36 mmHg lower home diastolic BP (P=.003). The association, however, was attenuated and became statistically nonsignificant with further adjustment for BMI. CONCLUSIONS: In this community-based sample of adults, higher daily habitual physical activity measured by a smartwatch was associated with a moderate, but statistically significant, reduction in home BP. Differences in BMI among study participants accounted for the majority of the observed association.

Adherence of Mobile App-Based Surveys and Comparison With Traditional Surveys: eCohort Study.

BACKGROUND: eCohort studies offer an efficient approach for data collection. However, eCohort studies are challenged by volunteer bias and low adherence. We designed an eCohort embedded in the Framingham Heart Study (eFHS) to address these challenges and to compare the digital data to traditional data collection. OBJECTIVE: The aim of this study was to evaluate adherence of the eFHS app-based surveys deployed at baseline (time of enrollment in the eCohort) and every 3 months up to 1 year, and to compare baseline digital surveys with surveys collected at the research center. METHODS: We defined adherence rates as the proportion of participants who completed at least one survey at a given 3-month period and computed adherence rates for each 3-month period. To evaluate agreement, we compared several baseline measures obtained in the eFHS app survey to those obtained at the in-person research center exam using the concordance correlation coefficient (CCC). RESULTS: Among the 1948 eFHS participants (mean age 53, SD 9 years; 57% women), we found high adherence to baseline surveys (89%) and a decrease in adherence over time (58% at 3 months, 52% at 6 months, 41% at 9 months, and 40% at 12 months). eFHS participants who returned surveys were more likely to be women (adjusted odds ratio [aOR] 1.58, 95% CI 1.18-2.11) and less likely to be smokers (aOR 0.53, 95% CI 0.32-0.90). Compared to in-person exam data, we observed moderate agreement for baseline app-based surveys of the Physical Activity Index (mean difference 2.27, CCC=0.56), and high agreement for average drinks per week (mean difference 0.54, CCC=0.82) and depressive symptoms scores (mean difference 0.03, CCC=0.77). CONCLUSIONS: We observed that eFHS participants had a high survey return at baseline and each 3-month survey period over the 12 months of follow up. We observed moderate to high agreement between digital and research center measures for several types of surveys, including physical activity, depressive symptoms, and alcohol use. Thus, this digital data collection mechanism is a promising tool to collect data related to cardiovascular disease and its risk factors.

The evolution of ependymin-related proteins.

BACKGROUND: Ependymins were originally defined as fish-specific secreted glycoproteins involved in central nervous system plasticity and memory formation. Subsequent research revealed that these proteins represent a fish-specific lineage of a larger ependymin-related protein family (EPDRs). EPDRs have now been identified in a number of bilaterian animals and have been implicated in diverse non-neural functions. The recent discoveries of putative EPDRs in unicellular holozoans and an expanded EPDR family with potential roles in conspecific communication in crown-of-thorns starfish suggest that the distribution and diversity of EPDRs is significantly broader than currently understood. RESULTS: We undertook a systematic survey to determine the distribution and evolution of EPDRs in eukaryotes. In addition to Bilateria, EPDR genes were identified in Cnidaria, Placozoa, Porifera, Choanoflagellatea, Filasterea, Apusozoa, Amoebozoa, Charophyta and Percolozoa, and tentatively in Cercozoa and the orphan group Malawimonadidae. EPDRs appear to be absent from prokaryotes and many eukaryote groups including ecdysozoans, fungi, stramenopiles, alveolates, haptistans and cryptistans. The EPDR family can be divided into two major clades and has undergone lineage-specific expansions in a number of metazoan lineages, including in poriferans, molluscs and cephalochordates. Variation in a core set of conserved residues in EPDRs reveals the presence of three distinct protein types; however, 3D modelling predicts overall protein structures to be similar. CONCLUSIONS: Our results reveal an early eukaryotic origin of the EPDR gene family and a dynamic pattern of gene duplication and gene loss in animals. This research provides a phylogenetic framework for the analysis of the functional evolution of this gene family.

Human Agency in Disaster Planning: A Systems Approach.

Current approaches to risk management place insufficient emphasis on the system knowledge available to the assessor, particularly in respect of the dynamic behavior of the system under threat, the role of human agents (HAs), and the knowledge available to those agents. In this article, we address the second of these issues. We are concerned with a class of systems containing HAs playing a variety of roles as significant system elements-as decisionmakers, cognitive agents, or implementers-that is, human activity systems. Within this family of HAS, we focus on safety and mission-critical systems, referring to this subclass as critical human activity systems (CHASs). Identification of the role and contribution of these human elements to a system is a nontrivial problem whether in an engineering context, or, as is the case here, in a wider social and public context. Frequently, they are treated as standing apart from the system in design or policy terms. Regardless of the process of policy definition followed, analysis of the risk and threats to such a CHAS requires a holistic approach, since the effect of undesirable, uninformed, or erroneous actions on the part of the human elements is both potentially significant to the system output and inextricably bound together with the nonhuman elements of the system. We present a procedure for identifying the potential threats and risks emerging from the roles and activity of those HAs, using the 2014 flooding in southwestern England and the Thames Valley as a contemporary example.

Patterns of Mood and Personality Factors and Associations With STI/HIV-Related Drug and Sex Risk Among African American Male Inmates.

BACKGROUND: Research on the association between antisocial personality disorder (ASPD) with comorbid mental disorders and sexually transmitted infection (STI)/HIV risk among inmates is scant despite the high prevalence of psychopathology and of STI/HIV in this population. METHODS: We used baseline data from Project DISRUPT, a cohort study conducted among incarcerated African American men (n = 207), to measure associations between ASPD and STI/HIV risk. We also conducted latent class analyses (LCAs) to identify subgroups defined by ASPD with comorbid stress, depression, and borderline personality disorder symptoms and measured associations between latent class membership and STI/HIV risk. RESULTS: Approximately 15% had ASPD and 39% reported depression. Controlling for sociodemographics, stress, and depression, ASPD was independently associated with illicit [AOR = 3.23, 95% confidence interval (CI): 1.18-8.87] and injection drug use (AOR: 5.49, 95% CI: 1.23-24.42) but not with sexual risk. LCAs suggested that those at high risk of ASPD were likely to experience co-morbid mental disorders. ASPD comorbid with these disorders was linked to drug and sex risk. CONCLUSIONS: STI/HIV prevention for inmates should incorporate diagnosis and treatment of ASPD and comorbid disorders, and interventions to address ASPD-related factors (e.g., impulsivity) that drive STI/HIV risk.

A randomized study of internet parent training accessed from community technology centers.

Behavioral parent training (BPT) has been shown to be efficacious to improve parenting skills for problematic interactions with adolescents displaying oppositional and antisocial behaviors. Some research suggests that support group curricula might be transferred to the Internet, and some studies suggest that other curriculum designs might also be effective. In this research, a BPT program for parents of at-risk adolescents was tested on the Internet in a randomized trial (N = 307) from computer labs at six community technology centers in or near large metropolitan areas. The instructional design was based on asynchronous scenario-based e-learning, rather than a traditional parent training model where presentation of course material builds content sequentially over multiple class sessions. Pretest to 30-day follow-up analyses indicated significant treatment effects on parent-reported discipline style (Parenting Scale, Adolescent version), child behavior (Eyberg Child Behavior Inventory), and on social cognitive theory constructs of intentions and self-efficacy. The effect sizes were small to medium. These findings suggest the potential to provide effective parent training programs on the Internet.

Stress, burnout, and the need for support: a survey of early career pharmacists.

BACKGROUND: Burnout of health professionals is of concern internationally and the pharmacy profession is no exception. The period of transition from University to autonomous practitioner is recognized to be challenging and these Early Career Pharmacists (ECPs), may be at increased risk of stress and burnout. OBJECTIVES: This study aimed to collect data on the current extent of self-identified stress and burnout, of ECPs, and to (i) identify contributing factors and (ii) identify strategies used to manage this stress. METHODS: This study was conducted in Aotearoa New Zealand and was based on a survey used previously in Australia. A national database was used to identify all pharmacists who had been registered for <10 years. The survey was emailed to each pharmacist, and was further advertised through social media platforms. RESULTS: A total of 1418 ECPs were identified and invited to participate, and responses were received by 416 of these. The majority of respondents were female (73%) and community pharmacy based (79%). A vast majority (89%) indicated that a normal working day was at least moderately stressful with 79% indicating a level of burnout. These reflect a combination of "external" stressors, i.e. negative patient interactions, staff shortages, and "internal" stressors, i.e. fear of making a mistake, adjustment from University to working life. CONCLUSIONS: Stress and burnout appear to be widespread in the ECPs, this study highlights areas of concern and potential support where efforts can be focussed to create a more sustainable working environment.

Career outlook and satisfaction in the presence of workload intensification-a survey of early career pharmacists.

BACKGROUND: The demographics of the pharmacy workforce is changing with an increased proportion of pharmacists less than 30 years old (early career pharmacists-ECPs). In parallel, the profession has experienced workload intensification and workforce attrition. It is important to understand ECPs career satisfaction to retain this section of the pharmacy profession. OBJECTIVES: This study aimed to collect data on the current career satisfaction of ECPs, and identify workplace factors that were most important to this group. Further, to use these findings to inform sector recommendations. METHODS: A steering group of ECPs in Aotearoa New Zealand developed a survey based on one used previously. An invitation email was sent to all pharmacists who had been registered for less than 10 years and were members of the Pharmaceutical Society of New Zealand. RESULTS: A total of 1418 ECPs were identified and invited to participate, and responses were received by 416 of these. While 90% believe that they are making a useful contribution to the health of their patients, over half are unhappy and discontented, with over a third dissatisfied with their careers. A large proportion (44%) were considering leaving the profession in the next 5 years. The top three factors for career satisfaction were ability to progress and learn new skills, the people they work with, and the remuneration. CONCLUSIONS: This study provides a starting point for understanding the current environment and level of dissatisfaction of young pharmacists. There are several areas of concern that need to be addressed if a strong vibrant viable pharmacy profession is to be achieved.

An Age-Period-Cohort Analysis of Cardiovascular Disease Mortality in the United States from 1999 to 2018.

BACKGROUND: Although cardiovascular disease mortality rates in the United States declined from the 1970s to 2010s, they have now plateaued. The independent effects of age, period, and birth year (cohort) on cardiovascular disease mortality have not previously been defined. METHODS: We used data from the Centers for Disease Control and Prevention's Wide-Ranging Online Data for Epidemiologic Research to examine the effects of age, period, and cohort on cardiovascular disease mortality among individuals aged 20-84 years from 1999 to 2018, prior to the onset of the coronavirus disease 2019 pandemic. Age effects were described as cardiovascular disease-related mortality rates in each 5-year age group adjusted for year of death (period) and year of birth (cohort). Period and cohort effects were quantified as adjusted rate ratios (RRs) comparing cardiovascular disease mortality rates in each period and cohort to the reference periods and reference cohort (ie, 1919 birth cohort), respectively. RESULTS: Between 1999 to 2018, there were 10,404,327 cardiovascular disease deaths among US adults. In each individual birth cohort, the age-specific cardiovascular disease mortality rates were stable between ages 20 through 39 years. Age-specific rates were higher for each year older between ages 40 through 84 years adjusting for period effects. The period cardiovascular disease mortality rates were lower in later periods (2004-2008 period RR 0.87, 95% confidence interval [CI] 0.85 to 0.88; 2009-2013 period RR 0.78, 95% CI 0.76 to 0.80) compared with the reference period (1999 to 2003) and plateaued thereafter. The cohort cardiovascular disease mortality rates were progressively lower in more recent birth cohorts (1924 birth cohort RR 0.85, 95% CI 0.83 to 0.87; 1974 birth cohort RR 0.29, 95% CI 0.27 to 0.32) compared with the reference cohort (1919 cohort) and plateaued thereafter. CONCLUSION: Although cardiovascular disease mortality rates declined rapidly among those born between 1919 and 1974, improvements plateaued in birth cohorts thereafter even adjusted for period effects.

Comprehensive sub-mitochondrial protein map of the parasitic protist Trypanosoma brucei defines critical features of organellar biology.

We have generated a high-confidence mitochondrial proteome (MitoTag) of the Trypanosoma brucei procyclic stage containing 1,239 proteins. For 337 of these, a mitochondrial localization had not been described before. We use the TrypTag dataset as a foundation and take advantage of the properties of the fluorescent protein tag that causes aberrant but fortuitous accumulation of tagged matrix and inner membrane proteins near the kinetoplast (mitochondrial DNA). Combined with transmembrane domain predictions, this characteristic allowed categorization of 1,053 proteins into mitochondrial sub-compartments, the detection of unique matrix-localized fucose and methionine synthesis, and the identification of new kinetoplast proteins, which showed kinetoplast-linked pyrimidine synthesis. Moreover, disruption of targeting signals by tagging allowed mapping of the mode of protein targeting to these sub-compartments, identifying a set of C-tail anchored outer mitochondrial membrane proteins and mitochondrial carriers likely employing multiple target peptides. This dataset represents a comprehensive, updated mapping of the mitochondrion.

Cardiac Structure and Function Phenogroups and Risk of Incident Heart Failure (from the Multi-ethnic Study of Atherosclerosis).

Indices of cardiac structure and function, such as left ventricular (LV) mass and ejection fraction, have been associated with risk of incident heart failure (HF), but the clinical relevance of data-driven grouping of a comprehensive set of cardiac parameters is unclear. In Multi-Ethnic Study of Atherosclerosis participants, latent class analysis was applied in the sample stratified by gender to define phenogroups on the basis of cardiovascular magnetic resonance imaging parameters of right ventricular and LV structure and function at baseline. Cox proportional hazard models in gender-stratified analyses were used to assess the association between phenogroup membership and risk of HF subtypes adjusting for potential confounders. In the 4,204 participants (mean age 61 ± 10 years, 53% women), the mean follow-up time was 14 ± 4 years for men and 15 ± 4 years for women. For both genders, 4 distinct phenogroups were identified: (1) ideal cardiac mechanics; (2) higher output/hypertrophied LV; (3) impaired ejection fraction/dilated LV; and (4) higher output/hyperdynamic (LV). Men in phenogroups 4 (hazard ratio [HR] 2.91, 95% confidence interval [CI] 1.60 to 5.31, p = 0.0005), 3 (HR 3.52, 95% CI 1.90 to 6.53, p <0.0001), and 2 (HR 3.49, 95% CI 1.94 to 6.28, p <0.0001) had higher rates of incident HF than did men in phenogroup 1, in fully adjusted models. No significant associations were found between phenogroup membership and incident HF in women. In conclusion, phenogroup membership based on cardiac structure and function in men was significantly associated with incident HF. Integration of cardiac magnetic resonance imaging variables may help identify differential risk for HF in men.

New strategies and therapies for the prevention of heart failure in high-risk patients.

Despite declines in total cardiovascular mortality rates in the United States, heart failure (HF) mortality rates as well as hospitalizations and readmissions have increased in the past decade. Increases have been relatively higher among young and middle-aged adults (<65 years). Therefore, identification of individuals HF at-risk (Stage A) or with pre-HF (Stage B) before the onset of overt clinical signs and symptoms (Stage C) is urgently needed. Multivariate risk models (e.g., Pooled Cohort Equations to Prevent Heart Failure [PCP-HF]) have been externally validated in diverse populations and endorsed by the 2022 HF Guidelines to apply a risk-based framework for the prevention of HF. However, traditional risk factors included in the PCP-HF model only account for half of an individual's lifetime risk of HF; novel risk factors (e.g., adverse pregnancy outcomes, impaired lung health, COVID-19) are emerging as important risk-enhancing factors that need to be accounted for in personalized approaches to prevention. In addition to determining the role of novel risk-enhancing factors, integration of social determinants of health (SDoH) in identifying and addressing HF risk is needed to transform the current clinical paradigm for the prevention of HF. Comprehensive strategies to prevent the progression of HF must incorporate pharmacotherapies (e.g., sodium glucose co-transporter-2 inhibitors that have also been termed the "statins" of HF prevention), intensive blood pressure lowering, and heart-healthy behaviors. Future directions include investigation of novel prediction models leveraging machine learning, integration of risk-enhancing factors and SDoH, and equitable approaches to interventions for risk-based prevention of HF.

Using psycho-behavioral phenotyping for overweight and obesity: Confirmation of the 6 factor questionnaire.

Objective: Questionnaires that assess dietary habits, eating behaviors, and relevant psychosocial constructs are routinely used in obesity research and clinical practice. The 6 factor questionnaire (6FQ) was previously developed as an assessment tool for psycho-behavioral phenotyping. The primary purpose of this study was to confirm and validate the original findings in a large diverse adult population. Methods: A total of 5399 self-selected participants (mean age of 48 ± 13 years and body mass index of 32 ± 8 kg/m2) completed the 6FQ online. The association between self-reported demographic data and 6FQ responses was assessed using linear regression models. Results: Mean factor score and odds ratio analyses consistently demonstrated a statistically significant relationship between factors and body weight even after adjusting for age, sex, and race/ethnicity. Conclusions: Although the study was correlational in design, the results demonstrate that the 6FQ, an instrument that represents multidimensional unhealthful lifestyle patterns associated with diet, physical activity, cognition, and self-perception worsen with increasing body weight. Psycho-behavioral phenotyping may be a useful approach when assessing and treating patients with obesity.

Attendance at Supervised Exercise Sessions and Walking Outcomes in Peripheral Artery Disease: Results From 2 Randomized Clinical Trials.

Background Supervised exercise therapy (SET) is the first-line therapy for walking impairment in peripheral artery disease (PAD). This study evaluated the association between attendance at SET and improved walking performance, compared with a control group, in PAD. Methods and Results Data from 2 randomized clinical trials of SET for PAD were combined. In each trial, participants were randomized to 3 times weekly supervised treadmill exercise or an attention control group for 6 months (maximum, 77 exercise sessions). Participants randomized to SET were categorized into tertiles, according to the proportion of exercise sessions they attended. Results adjusted for age, sex, race, baseline walking performance, comorbidities, and other potential confounders. A total of 272 participants with PAD (mean age, 67.9±9.3 years; 44% women; 61% Black race) were included. For participants randomized to SET, tertiles of attendance rates at exercise sessions were as follows: 11% to 68% (N=45), 69% to <85% (N=46), and ≥85% (N=46). Compared with control, mean improvement in 6-minute walk was significantly greater in each SET tertile: mean (95% CI) for tertile 1, 27.9 m (1.3-54.4 m; P=0.04), tertile 2, 38.2 m (12.2-64.2 m; P=0.001), and tertile 3, 56.9 m (29.9-83.8 m; P<0.0001). Among participants randomized to SET, greater SET attendance was associated with greater improvement in 6-minute walk distance (overall P for trend=0.025). Compared with control, improvement in maximal treadmill walking time was greater in each SET attendance tertile: tertile 1 (3.3 minutes [95% CI, 1.7-4.8 minutes]; P<0.0001), tertile 2 (3.8 minutes [95% CI, 2.3-5.3 minutes]; P<0.0001), and tertile 3 (5.4 minutes [95% CI, 3.9-7.0 minutes]; P:<0.0001). Among participants randomized to SET, greater attendance at SET was not significantly associated with greater improvement in maximal treadmill walking time (overall P for trend=0.064). Conclusions Among people with PAD randomized to SET, better attendance at exercise sessions was associated with significantly greater 6-minute walk improvement. Among all participants with PAD, even relatively low SET attendance was associated with significantly greater improvement in walking performance, compared with a control group who did not exercise. Registration URL: https://clinicaltrials.gov/ct2/show/NCT01408901. URL: https://clinicaltrials.gov/ct2/show/NCT00106327.

Diplonemids - A Review on "New" Flagellates on the Oceanic Block.

Diplonemids are a group of flagellate protists, that belong to the phylum Euglenozoa alongside euglenids, symbiontids and kinetoplastids. They primarily inhabit marine environments, though are also found in freshwater lakes. Diplonemids have been considered as rare and unimportant eukaryotes for over a century, with only a handful of species described until recently. However, thanks to their unprecedented diversity and abundance in the world oceans, diplonemids now attract increased attention. Recent improvements in isolation and cultivation have enabled characterization of several new genera, warranting a re-examination of all available knowledge gathered so far. Here we summarize available data on diplonemids, focusing on the recent advances in the fields of diversity, ecology, genomics, metabolism, and endosymbionts. We illustrate the life stages of cultivated genera, and summarise all reported interspecies associations, which in turn suggest lifestyles of predation and parasitism. This review also includes the latest classification of diplonemids, with a taxonomic revision of the genus Diplonema. Ongoing efforts to sequence various diplonemids suggest the presence of large and complex genomes, which correlate with the metabolic versatility observed in the model species Paradiplonema papillatum. Finally, we highlight its successful transformation into one of few genetically tractable marine protists.