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BackgroundBoth elevated and low resting heart rates are associated with atrial fibrillation (AF), suggesting a U-shaped relationship. However, evidence for a U-shaped causal association between genetically-determined resting heart rate and incident AF is limited. We investigated potential directional changes of the causal association between genetically-determined resting heart rate and incident AF.Method and resultsSeven cohorts of the AFGen consortium contributed data to this meta-analysis. All participants were of European ancestry with known AF status, genotype information, and a heart rate measurement from a baseline electrocardiogram (ECG). Three strata of instrumental variable-free resting heart rate were used to assess possible non-linear associations between genetically-determined resting heart rate and the logarithm of the incident AF hazard rate: 75 beats per minute (bpm). Mendelian randomization analyses using a weighted resting heart rate polygenic risk score were performed for each stratum. We studied 38,981 individuals (mean age 59±10 years, 54% women) with a mean resting heart rate of 67±11 bpm. During a mean follow-up of 13±5 years, 4,779 (12%) individuals developed AF. A U-shaped association between the resting heart rate and the incident AF-hazard ratio was observed. Genetically-determined resting heart rate was inversely associated with incident AF for instrumental variable-free resting heart rates below 65 bpm (hazard ratio for genetically-determined resting heart rate, 0.96; 95% confidence interval, 0.94-0.99; p = 0.01). Genetically-determined resting heart rate was not associated with incident AF in the other two strata.ConclusionsFor resting heart rates below 65 bpm, our results support an inverse causal association between genetically-determined resting heart rate and incident AF.

Abstract Background Biologic (b) and targeted synthetic (ts) disease-modifying anti-rheumatic drugs (DMARDs) used in the management of rheumatoid arthritis (RA) target inflammatory pathways implicated in the pathogenesis of multiple myeloma (MM). It is unknown whether use of b/tsDMARDs affects the incidence of MM. Methods In this cohort study using Veterans Health Administration (VHA) data, we identified Veterans newly diagnosed with RA from 1/1/2002 to 12/31/2018 using diagnostic codes and medication fills. DMARD exposure was categorized as follows: conventional synthetic (cs)DMARDs; bDMARDs, which included tumor necrosis factor inhibitors (TNFi), non-TNFi; and a tsDMARD, tofacitinib. A Cox proportional hazards model with time-varying exposure was used to estimate the hazard ratio for developing MM among those who received b/tsDMARD medications relative to b/tsDMARD-naïve persons. Results 27,540 veterans with RA met eligibility criteria of whom 8322 (30%) took a b/tsDMARD during follow-up. There were 77 incident cases of MM over 192,000 person-years of follow-up. The age-adjusted incidence rate (IR) of MM among b/tsDMARD-naïve patients was 0.37 (95% CI 0.28–0.49) per 1000 person-years and 0.42 among current or former b/tsDMARD users (95% CI 0.25–0.65). Adjusting for age and other demographic characteristics, the hazard ratio for MM associated with use of b/tsDMARDs was 1.32 (95% CI 0.78, 2.26). Conclusion In this study of Veterans with RA, the rate of MM did not differ between b/tsDMARD and csDMARD users. The relatively short duration of follow-up and few events limited our power to detect treatment-related differences in MM risk.

Abstract The immunologic consequences of using bactericidal versus bacteriostatic antibiotic treatments are unclear. We observed a bacteriostatic (growth halting) treatment was more protective than a bactericidal (bacteria killing) treatment in a murine peritonitis model. To understand this unexpected difference, we compared macrophage responses to bactericidal treated bacteria or bacteriostatic treated bacteria. We found that Gram-negative bacteria treated with bactericidal drugs induced more proinflammatory cytokines than those treated with bacteriostatic agents. Bacterial DNA – released only by bactericidal treatments – exacerbated inflammatory signaling through TLR9. Without TLR9 signaling, the in vivo efficacy of bactericidal drug treatment was rescued. This demonstrates that antibiotics can act in important ways distinct from bacterial inhibition: like causing treatment failure by releasing DNA that induces excessive inflammation. These data establish a novel link between how an antibiotic affects bacterial physiology and subsequent immune system engagement, which may be relevant for optimizing treatments to simultaneously clear bacteria and modulate inflammation.

Abstract Background Night shift workers face increased risk of cardiovascular disease (CVD) compared to non-shift workers. Evidence supports on-shift napping and regular non-invasive monitoring of endothelial function for risk mitigation, yet neither strategy is widely used. Methods We evaluated the feasibility of non-invasive assessment of peripheral arterial tone (PAT) to assess the effect of napping during simulated night shift work on endothelial function. We used a single-site, randomized crossover trial of simulated night shift work with a 45-min nap condition versus a control, no-nap condition (ClinicalTrials.gov NCT05436951). Results The primary outcome was the number of participants with ≥ 70% of endothelial function assessments. Secondary outcomes included mean reactive hyperemia index (RHI), BP, and cognitive performance with the brief psychomotor vigilance task (PVT-B). Of the 10 consented, 9 completed both conditions. All participants exceeded feasibility benchmarks. Mean RHI did not differ by nap condition, and the delta from pre- to post measure did not differ (difference in delta = − 0.26, 95% CI − 1.09, 0.58). Hourly PVT-B assessments from 19:00 to 07:00 h did not differ by nap condition. Compared to pre-nap measures, cognitive performance on the PVT-B was poorest at + 0 min post-nap. Conclusion Our findings can inform larger studies evaluating the effects of night shift work and napping on endothelial function. Trial registrations ClinicalTrials.gov (NCT05436951, registered on June 23, 2022).

Wildfires are a significant agent of disturbance in forests and highly sensitive to climate change. Short-interval fires and high severity (mortality-causing) fires in particular, may catalyze rapid and substantial ecosystem shifts by eliminating woody species and triggering conversions from forest to shrub or grassland ecosystems. Modeling and fine-scale observations suggest negative feedbacks between fire and fuels should limit reburn prevalence as overall fire frequency rises. However, while we have good information on reburning patterns for individual fires or small regions, the validity of scaling these conclusions to broad regions like the US West remains unknown. Both the prevalence of reburning and the strength of feedbacks on likelihood of reburning over differing timescales have not been documented at the regional scale. Here we show that while there is a strong negative feedback for very short reburning intervals throughout wildland forests of the Western US, that feedback weakens after 10–20 years. The relationship between reburning intervals and drought diverges depending on location, with coastal systems reburning quicker (e.g. shorter interval between fires) in wetter conditions and interior forests in drier. This supports the idea that vegetation productivity—primarily fine fuels that accumulate rapidly (

Abstract The significant interest in actinide bonding has recently focused on novel compounds with exotic oxidation states. However, the difficulty in obtaining relevant high-quality experimental data, particularly for low-valent actinide compounds, prevents a deeper understanding of 5f systems. Here we show X-ray absorption near-edge structure (XANES) measurements in the high-energy resolution fluorescence detection (HERFD) mode at the uranium M4 edge for the UIII and UIV halides, namely UX3 and UX4 (X = F, Cl, Br, I). The spectral shapes of these two series exhibit clear differences, which we explain using electronic structure calculations of the 3d-4f resonant inelastic X-ray scattering (RIXS) process. To understand the changes observed, we implemented crystal field models with ab initio derived parameters and investigated the effect of reducing different contributions to the electron-electron interactions involved in the RIXS process. Our analysis shows that the electron-electron interactions weaken as the ligand changes from I to F, indicative of a decrease in ionicity both along and between the UX3 and UX4 halide series.

Objective Identification of characteristics associated with active disease in juvenile idiopathic arthritis (JIA) could inform early disease treatment strategies. This study evaluated characteristics associated with active disease at 12 and 24 months after JIA diagnosis in the era in which biologic disease‐modifying antirheumatic drugs (DMARDs) became available for JIA. Methods This single‐center retrospective study from 2004 through 2018 assessed characteristics associated with active nonsystemic categories of JIA at 12 and 24 months after diagnosis. Relative prevalence (RP) of disease activity was evaluated in relation to prespecified characteristics. Using RP, the effect of increasing biologic DMARD availability on these predictors was assessed at 12 months. Results A total of 1,151 patients with JIA were included. At 12 months, a 40% to 45% higher point prevalence of active disease was noted in older children (>5 years). Patients with active disease at 3 months had a greater prevalence of active disease at 12 months (RP 1.5, 95% confidence interval [CI] 1.2–1.8) and 24 months (RP 1.3, 95% CI 1–1.6). Compared to oligoarticular JIA, polyarticular RF‐negative, psoriatic, and enthesitis‐related JIA had a greater prevalence of active disease at 12 and 24 months. At 24 months, a greater prevalence of active disease was observed in children ≥10 years. RP of active disease was 25% lower in the late cohort (2013–2018) than in the earliest cohort (2004–2008; RP 0.75, 95% CI 0.62–0.92) when more biologic medications were available, but disease activity predictors were broadly similar over time. Conclusion Patients with JIA with active disease at 12 and 24 months were older at diagnosis, categorized as polyarticular RF‐negative, psoriatic, or enthesitis‐related JIA. Active disease at 3 months after diagnosis was associated with worse outcomes at 12 and 24 months.

Abstract Disruptions of the eukaryotic plasma membrane due to chemical and mechanical challenges are frequent and detrimental and thus need to be repaired to maintain proper cell function and avoid cell death. However, the cellular mechanisms involved in wound resealing and restoration of homeostasis are diverse and contended. Here, it is shown that clathrin‐mediated endocytosis is induced at later stages of plasma membrane wound repair following the actual resealing of the wound. This compensatory endocytosis occurs near the wound, predominantly at sites of previous early endosome exocytosis which is required in the initial stage of membrane resealing, suggesting a spatio‐temporal co‐ordination of exo‐ and endocytosis during wound repair. Using cytoskeletal alterations and modulations of membrane tension and membrane area, membrane tension is identified as a major regulator of the wounding‐associated exo‐ and endocytic events that mediate efficient wound repair. Thus, membrane tension changes are a universal trigger for plasma membrane wound repair modulating the exocytosis of early endosomes required for resealing and subsequent clathrin‐mediated endocytosis acting at later stages to restore cell homeostasis and function.

Abstract Glucose plays critical roles in many human body functions, above all as a source of energy. Abnormal levels of glucose are correlated to different diseases, importantly including diabetes. As such, quantification of glucose levels in body fluids is essential for health monitoring. Blood tests and, more recently, portable interstitial fluid tests, currently represent the benchmarks for glucose detection. Inconvenient invasive methods such as blood tests pose burdens on both patients and the healthcare system. In this review, noninvasive approaches to measure glucose levels in the human body are discussed, utilizing saliva as an alternative to conventional blood samples. Techniques explored and with the potential to enhance accuracy and their associated challenges are discussed.

Abstract Introduction A three-pronged approach to acne treatment combining an antibiotic, antimicrobial, and retinoid may be more efficacious than single/double treatments while potentially reducing antibiotic resistance. This study evaluated the efficacy and safety of the first fixed-dose, triple-combination topical acne product, clindamycin 1.2%/adapalene 0.15%/benzoyl peroxide (BPO) 3.1% gel (CAB) using pooled phase 3 data. Methods In two identical phase 3 (N = 183; N = 180), double-blind, 12-week studies, participants aged ≥ 9 years with moderate-to-severe acne were randomized 2:1 to receive once-daily CAB or vehicle gel. Endpoints included ≥ 2-grade reduction from baseline in Evaluator’s Global Severity Score and clear/almost clear skin (treatment success) and least-squares mean percent change from baseline in acne lesion counts. Treatment-emergent adverse events (TEAEs) and cutaneous safety/tolerability were evaluated. Results At week 12, 50.0% of participants achieved treatment success with CAB versus 22.6% with vehicle gel (P 70% reductions in inflammatory and noninflammatory lesions at week 12 (77.9% and 73.0%, respectively), which were significantly greater than vehicle (57.9% and 48.2%; P

ABSTRACT Contact-dependent hemolysins are virulence factors in a number of human pathogens, including Helicobacter pylori, Rickettsia typhi, Bartonella bacilliformis, Mycobacterium tuberculosis, entero-invasive Escherichia coli, and Shigella. Here we demonstrate that Neisseria gonorrhoeae produces an outer membrane protein, phospholipase A, that exhibits contact-dependent lytic activity on host cell membranes. This enzyme can lyse human erythrocytes over a 3-day period, whereas a phospholipase A mutant cannot. We demonstrated phospholipase A activity in the parent strain but not in two, independent phospholipase A mutants. A gene for phospholipase A, pldA (hereafter referred to as pla to avoid confusion with the gene for phospholipase D, pld), is present in all sequenced gonococcal strains. Fluid phase, hemolytic activity assays showed that 25 of 29 gonococcal strains tested had hemolytic activity greater than 50% of the positive control. In support of PLA as a gonococcal outer membrane protein, supernatants from 24-, 48-, and 72-h cultures of N. gonorrhoeae strain 1291 did not contain hemolysin activity, and a monoclonal antibody specific for gonococcal phospholipase A failed to detect the enzyme in these supernatants. The organism must be viable for lysis to occur, and the inclusion of EDTA in the media removes all activity. Our studies have shown that a phospholipase A mutant has significantly reduced survival in human neutrophils and primary human cervical epithelial cells compared to the parent gonococcal strain after 3 h of incubation. Collectively, our data demonstrate that gonococcal PLA lyses host cell membranes, which is important for intracellular survival.IMPORTANCEIntracellular survival is crucial to the success of Neisseria gonorrhoeae as a human pathogen. Multiple factors contribute to the intracellular survival of gonococci, including the ability to prohibit apoptosis of the epithelial cell the organism invades and mechanisms to evade host innate defense systems. The role of phospholipase A (PLA), an outer membrane protein, is important as it disrupts the host vacuolar and phagolysosomal membranes, preventing the effective delivery of innate immune factors that normally restrict organism growth within human cells. After cell entry, PLA disrupts the integrity of these host cell membranes, allowing the gonococcus to live free within disrupted vacuoles where it pilfers host cell nutrients that enable its survival and replication. A vaccine or drug that could neutralize PLA activity would disrupt the intracellular survival of the gonococcus.

Abstract Cellular senescence leads to the functional decline of regenerative cells such as mesenchymal stromal/stem cells (MSCs), which gives rise to chronic conditions and contributes to poor cell therapy outcomes. Aging tissues are associated with extracellular matrix (ECM) dysregulation, including loss of elastin. However, the role of the ECM in modulating senescence is underexplored. In this work, it is shown that tropoelastin, the soluble elastin precursor, is not only a marker of young MSCs but also actively preserves cell fitness and delays senescence during replicative aging. MSCs briefly exposed to tropoelastin exhibit upregulation of proliferative genes and concurrent downregulation of senescence genes. The seno‐protective benefits of tropoelastin persist during continuous, long‐term MSC culture, and significantly extend the MSC replicative lifespan. Tropoelastin‐expanded MSCs further maintain youth‐associated phenotype and function compared to age‐matched controls, including preserved clonogenic potential, minimal senescence‐associated beta‐galactosidase activity, maintained cell sizes, reduced expression of senescence markers, suppressed secretion of senescence‐associated factors, and increased production of youth‐associated proteins. This work points to the utility of exogenously‐supplemented tropoelastin for manufacturing MSCs that robustly maintain regenerative potential with age. It further reveals the active role of classical structural ECM proteins in driving cellular age‐associated fitness, potentially leading to future interventions for aging‐related pathologies.

Objectives Only 5–8% of adults with cancer participate in cancer clinical trials (CCTs), with even lower rates among underrepresented groups. Improving oncologists’ communication skills may enhance the frequency and quality of their discussions with patients about CCTs, consequently increasing participation. However, little is known about interest in or presence of CCT-related communication training during Hematology-Oncology (Hem-Onc) fellowships. This study aimed to describe, from the perspective of Hem-Onc fellowship program directors (PDs): (1) the current landscape of CCT education for Hem-Onc fellows; (2) the acceptability and feasibility of implementing a CCT communication skills workshop for Hem-Onc fellows. Methods We used an explanatory sequential mixed-methods approach. PDs were surveyed and interviewed about their graduate medical education (GME) programs’ current CCT curriculum, training challenges, fellows’ CCT knowledge and CCT communication skills, and preferences for a CCT communication workshop. Results PDs were surveyed (n = 40) and interviewed (n = 12). PDs reported that their institutions prioritize CCT accrual (M = 4.58, SD = .78; 1-5 scale, 5 = “Strongly Agree”) and clinical research training (M = 4.20, SD = .85). CCT skills that programs least often addressed were how to (1) discuss CCTs with newly diagnosed patients, (2) talk to patients about CCTs when none are available, and (3) help patients find CCTs at other institutions. PDs were interested in a CCT communication workshop for fellows (“yes” = 67.5%, “maybe” = 32.5%) and said training would be feasible (M = 4.28, SD = .78) and useful (M = 4.47, SD = .78). Qualitative results described programs’ current approaches to CCT education and insights about developing and implementing CCT communication training. Conclusions There is a clear need to improve CCT communication skills training in Hem-Onc fellowship programs and to implement and scale such training to increase CCT participation, especially among diverse patient populations. Furthermore, Hem-Onc GME PDs view such training as feasible and useful.

We present a scheme for cooling a vibrational mode of a magnetic molecular nanojunction by a spin-polarized charge current upon exploiting the interaction between its magnetic moment and the vibration. The spin-polarized charge current polarizes the magnetic moment of the nanoisland, thereby lowering its energy. A small but finite coupling between the vibration and the magnetic moment permits a direct exchange of energy such that vibrational energy can be transferred into the magnetic state. For positive bias voltages, this generates an effective cooling of the molecular vibrational mode. We determine parameter regimes for the cooling of the vibration to be optimal. Although the flowing charge current inevitably heats up the vibrational mode via Ohmic energy losses, we show that due to the magnetomechanical coupling, the vibrational energy (i.e, the effective phonon temperature) can be lowered below 50% of its initial value, when the two leads are polarized anti-parallel. In contrast to the cooling effect for positive bias voltages, net heating of the vibrational mode occurs for negative bias voltages. The cooling effect is enhanced for a stronger anti-parallel magnetic polarization of the leads, while the heating is stronger for a larger parallel polarization. Yet, dynamical cooling is also possible with parallel lead alignments when the two tunneling barriers are asymmetric.

Familial dysautonomia (FD) is a genetic disease of the autonomous and sensory nervous systems. Severe gastro-oesophageal reflux is common and one of the major complications. Some patients with FD develop megaoesophagus. Oesophageal malfunction, accompanied by oesophageal food and secretion retention, results in recurrent aspiration and other severe respiratory complications. Through a traditional case report, we wish to show how reverse tubing of the oesophagus can lead to significant symptomatic improvement in these patients. Moreover, this technique can serve as an alternative treatment for other oesophageal motility disorders.

Abstract The hepatic content of amyloid beta (Aβ) decreases drastically in human and rodent cirrhosis highlighting the importance of understanding the consequences of Aβ deficiency in the liver. This is especially relevant in view of recent advances in anti‐Aβ therapies for Alzheimer's disease (AD). Here, it is shown that partial hepatic loss of Aβ in transgenic AD mice immunized with Aβ antibody 3D6 and its absence in amyloid precursor protein (APP) knockout mice (APP‐KO), as well as in human liver spheroids with APP knockdown upregulates classical hallmarks of fibrosis, smooth muscle alpha‐actin, and collagen type I. Aβ absence in APP‐KO and deficiency in immunized mice lead to strong activation of transforming growth factor‐β (TGFβ), alpha secretases, NOTCH pathway, inflammation, decreased permeability of liver sinusoids, and epithelial‐mesenchymal transition. Inversely, increased systemic and intrahepatic levels of Aβ42 in transgenic AD mice and neprilysin inhibitor LBQ657‐treated wild‐type mice protect the liver against carbon tetrachloride (CCl4)‐induced injury. Transcriptomic analysis of CCl4‐treated transgenic AD mouse livers uncovers the regulatory effects of Aβ42 on mitochondrial function, lipid metabolism, and its onco‐suppressive effects accompanied by reduced synthesis of extracellular matrix proteins. Combined, these data reveal Aβ as an indispensable regulator of cell–cell interactions in healthy liver and a powerful protector against liver fibrosis.

Abstract Tissue homeostasis and disease states rely on the formation of new blood vessels through angiogenic sprouting, which is tightly regulated by the properties of the surrounding extracellular matrix. While physical cues, such as matrix stiffness or degradability, have evolved as major regulators of cell function in tissue microenvironments, it remains unknown whether and how physical cues regulate endothelial cell migration during angiogenesis. To investigate this, a biomimetic model of angiogenic sprouting inside a tunable synthetic hydrogel is created. It is shown that endothelial cells sense the resistance of the surrounding matrix toward proteolytic cleavage and respond by adjusting their migration phenotype. The resistance cells encounter is impacted by the number of covalent matrix crosslinks, crosslink degradability, and the proteolytic activity of cells. When matrix resistance is high, cells switch from a collective to an actomyosin contractility‐dependent single cellular migration mode. This switch in collectivity is accompanied by a major reorganization of the actin cytoskeleton, where stress fibers are no longer visible, and F‐actin aggregates in large punctate clusters. Matrix resistance is identified as a previously unknown regulator of angiogenic sprouting and, thus, provides a mechanism by which the physical properties of the matrix impact cell migration modes through cytoskeletal remodeling.

Abstract Introduction Despite the importance of clinical trial participation among cancer patients, few participate—and even fewer patients from ethnic and racial minoritized groups. It is unclear whether suggested approaches to increase accrual are successful. We conducted a scoping review to identify evidence‐based approaches to increase participation in cancer treatment clinical trials that demonstrated clear increases in accrual. Notably, more stringent than other published reviews, only those studies with comparison data to measure a difference in accrual rates were included. Methods We searched PubMed/MEDLINE, Embase, CINAHL, and Web of Science for English‐language articles published from January 1, 2012, to August 8, 2022. Studies were included if they were conducted in the United States, described single or multicomponent interventions, and provided data to measure accrual relative to baseline levels or that compared accrual rates with other interventions. Results Sixteen articles were included: six with interventions addressing patient barriers, two addressing provider barriers, seven describing institutional change, and one describing policy change. Key themes emerged, such as a focus on patient education, cultural competency, and building the capacity of clinics. Few studies provide comparative accrual data, making it difficult to identify with certainty any effective, evidence‐based approaches for increasing accrual. Some patient‐ and system‐level interventions studies showed modest increases in accrual primarily through pre‐post measurement. Conclusion Despite an extensive body of literature about the barriers that impede cancer treatment trial accrual, along with numerous recommendations for how to overcome these barriers, results reveal surprisingly little evidence published in the last 10 years on interventions that increase accrual relative to baseline levels or compared with other interventions. As clinical trials are a primary vehicle through which we improve cancer care, it is critical that evidence‐based approaches are used to inform all efforts to increase accrual. Strategies for increasing participation in cancer clinical trials must be developed and rigorously evaluated so that these strategies can be disseminated, participation in trials can increase and become more equitable, and trial results can become more generalizable.

Creativity can be measured with a variety of methods including self-reports, others reports, and ability tests. While typical self-reports are best understood as weak proxies of creativity, biographical reports that assess previous creative activities seem more promising. Drawbacks of such measures -- including skewed item distributions, a lack of measurement invariance across sex, and low convergent validity with established measures of creativity (such as divergent thinking tasks) -- might be attributable to issues of item sampling. Therefore, we use a meta-heuristic algorithm ("Ant Colony Optimization") to develop a short-scale of creative activities that a) maintains coverage of the original domains, b) provides good model fit, c) has good reliability, d) demonstrates balanced item difficulty, e) is measurement invariant across sex, and f) possesses convergent validity with divergent thinking. For an 8-item short version of the original "Inventory of Creative Activities and Achievements," we identified an item set that satisfies the most pre-specified criteria using data from N = 296 adults. In a replication sample (N = 327) we replicate the findings and report good model fit, good reliability, adequate item difficulty, and measurement invariance across sex. We derive recommendations on the measurement of creative activities, and discuss implications for understanding creativity in general.

OBJECTIVES: This study aimed to test the effect of a 30-minute nap versus a 2-hour nap opportunity taken during a simulated night shift on performance, fatigue, sleepiness, mood, and sleep at the end of shift and during post-night shift recovery. METHODS: We conducted a randomized crossover trial of three nap conditions (30-minute, 2-hour, and no-nap) during 12-hour simulated night shifts. We tested for differences in performance, fatigue, sleepiness, mood, and sleep during in-lab and at-home recovery. Performance was measured with the Brief Psychomotor Vigilance Test (PVT-B). Subjective ratings were assessed with single-item surveys. RESULTS: Twenty-eight individuals consented to participate [mean age 24.4 (standard deviation 7.2) years; 53.6% female; 85.7% Emergency Medical Services clinicians]. PVT-B false starts at the end of the 12-hour night shift (at 07:00 hours) and at the start of in-lab recovery (08:00 hours) were lower following the 2-hour nap versus other conditions (P0.05). Sleepiness, fatigue, and some mood states were lower at most hourly assessments during the in-lab recovery period following the 2-hour nap condition compared to the other conditions. Sleep during recovery did not differ by duration of night shift nap. CONCLUSIONS: A 2-hour nap opportunity versus a 30-minute or no-nap opportunity is beneficial for performance, alertness, and mood post-night shift. No differences were detected in sleep during recovery.

Background/Objectives: Bone morphogenetic protein and activin membrane-bound inhibitor (BAMBI) is an antagonist of transforming growth factor (TGF)-β type 1 signaling. BAMBI functions as an anti-fibrotic protein and exerts pro- as well as anti-cancerogenic activities. Our study aimed to correlate hepatocyte BAMBI protein levels in hepatocellular carcinoma (HCC) with T stage, lymph node invasion, vessel invasion, grading, tumor size and Union for International Cancer Control (UICC) stage, as well as with liver inflammation and fibrosis stages. Methods: Hepatocyte BAMBI protein expression was assessed by immunohistochemistry in HCC tissues of 320 patients and non-tumor tissues of 51 patients. Results: In the HCC tissues of the whole cohort and sex-specific analysis, BAMBI protein was not related to T stage, vessel invasion, lymph node invasion, histologic grade, UICC stage and tumor size. Accordingly, BAMBI was not associated with overall survival, recurrence-free and metastasis-free survival. BAMBI protein levels in tumor and non-tumor tissues were not related to inflammation and fibrosis grade. BAMBI protein levels in HCC tissues and non-tumor tissues from HCC patients, which were analyzed by immunoblot in a small cohort and by immunohistochemistry in the tissues of patients described above, were similar. Notably, BAMBI protein was low-abundant in HCC tissues of hepatitis C virus (HCV) compared to hepatitis B virus (HBV)-infected patients with comparable disease severity. Immunoblot analysis revealed reduced BAMBI protein in non-tumor tissues of patients with HCV in comparison to patients with HBV and normal human liver tissues. Conclusions: In summary, this analysis showed that hepatocyte BAMBI protein levels of patients with HCC are related to HCV infection rather than the severity of the underlying liver disease and cancer staging.

The mechanisms underlying the remarkable capacity of the liver to regenerate are still not completely understood. Particularly, the cross-talk between cytokines and cellular components of the process is of utmost importance because they represent potential avenues for diagnostics and therapeutics. WNT1-inducible-signaling pathway protein 1 (WISP1) is a cytokine member of the CCN family, a family of proteins that play many different roles in liver pathophysiology. WISP1 also belongs to the earliest and strongest upregulated genes in mouse livers after CCl4 intoxication and has recently been shown to be secreted by tumor cells and to bind to type 1 collagen to cause its linearization in vitro and in tumor tissue in vivo. We show that WISP1 expression is strongly induced by TGFβ, a critical cytokine in wound healing processes. Additionally, secretion of WISP1 protein by hepatic stellate is increased in cells upon TGFβ stimulation (~seven-fold increase). Furthermore, WISP1 facilitates the migration of mouse hepatic stellate cells through collagen in vitro. However, in WISP1 knockout mice, no difference in stellate cell accumulation in damaged liver tissue and no influence on fibrosis was obtained, probably because the knockout of WISP1 was compensated by other factors in vivo.

Abstract Taurine-respiring gut bacteria produce H2S with ambivalent impact on host health. We report the isolation and ecophysiological characterization of a taurine-respiring mouse gut bacterium. Taurinivorans muris strain LT0009 represents a new widespread species that differs from the human gut sulfidogen Bilophila wadsworthia in its sulfur metabolism pathways and host distribution. T. muris specializes in taurine respiration in vivo, seemingly unaffected by mouse diet and genotype, but is dependent on other bacteria for release of taurine from bile acids. Colonization of T. muris in gnotobiotic mice increased deconjugation of taurine-conjugated bile acids and transcriptional activity of a sulfur metabolism gene-encoding prophage in other commensals, and slightly decreased the abundance of Salmonella enterica, which showed reduced expression of galactonate catabolism genes. Re-analysis of metagenome data from a previous study further suggested that T. muris can contribute to protection against pathogens by the commensal mouse gut microbiota. Together, we show the realized physiological niche of a key murine gut sulfidogen and its interactions with selected gut microbiota members.

Abstract A challenging task to understand health and disease-related microbiome signatures is to move beyond descriptive community-level profiling towards disentangling microbial interaction networks. Using a synthetic gut bacterial community, we aimed to study the role of individual members in community assembly, identify putative keystone species and test their influence across different environments. Single-species dropout experiments reveal that bacterial strain relationships strongly vary not only in different regions of the murine gut, but also across several standard culture media. Mechanisms involved in environment-dependent keystone functions in vitro include exclusive access to polysaccharides as well as bacteriocin production. Further, Bacteroides caecimuris and Blautia coccoides are found to play keystone roles in gnotobiotic mice by impacting community composition, the metabolic landscape and inflammatory responses. In summary, the presented study highlights the strong interdependency between bacterial community ecology and the biotic and abiotic environment. These results question the concept of universally valid keystone species in the gastrointestinal ecosystem and underline the context-dependency of both, keystone functions and bacterial interaction networks.

Meiotic sex chromosome inactivation (MSCI) is a critical feature of meiotic prophase I progression in males. While the ATR kinase and its activator TOPBP1 are key drivers of MSCI within the specialized sex body (SB) domain of the nucleus, how they promote silencing remains unclear given their multifaceted meiotic functions that also include DNA repair, chromosome synapsis, and SB formation. Here we report a novel mutant mouse harboring mutations in the TOPBP1-BRCT5 domain. Topbp1B5/B5 males are infertile, with impaired MSCI despite displaying grossly normal events of early prophase I, including synapsis and SB formation. Specific ATR-dependent events are disrupted, including phosphorylation and localization of the RNA:DNA helicase Senataxin. Topbp1B5/B5 spermatocytes initiate, but cannot maintain ongoing, MSCI. These findings reveal a non-canonical role for the ATR-TOPBP1 signaling axis in MSCI dynamics at advanced stages in pachynema and establish the first mouse mutant that separates ATR signaling and MSCI from SB formation.

Rationale: Parents of a child or adolescent (CA) or young adult (YA) diagnosed with a hematologic cancer often face uncertainty. Managing uncertainty is critical to reduce the psychosocial burden of illness-related stressors. Objective: This study sought to identify: 1) sources of uncertainty among parents of a child diagnosed with a hematologic cancer, 2) strategies used by parents to manage uncertainty, and 3) clinicians' responses to parents' online information-seeking approach to managing uncertainty. Methods: Parents of CAs/YAs diagnosed with a hematologic cancer within the past 1–18 months and living in the U.S. participated in an in-depth, semi-structured phone interview (n = 20). Data were analyzed thematically. Results: Parents reported uncertainty about treatment (options, efficacy, and side effects or risks) and uncertainty about the future (recurrence, whether worry would subside, and how to approach the child's future). Parents managed uncertainty by seeking information online, talking to clinicians, and joining support groups. Clinicians' responses to online information-seeking were described as supportive and unsupportive. Conclusion: Parents described struggling with uncertainty across the cancer continuum (from primary treatment to survivorship). Parents' psychosocial health may benefit from individual and systems level interventions that help address and manage uncertainty, especially interventions focusing on parent caregiver-clinician communication.

Introduction: Group well-child care (GWCC) is an alternative to traditional pediatric well-child care designed to increase parental social support and peer learning. This mixed methods study explored the adaptation and implementation of GWCC to a virtual format during coronavirus disease 2019 (COVID-19 pandemic) among Spanish-speaking Latino immigrant families. Method: Interviews were conducted with eight providers and 10 mothers from May through September 2020. Qualitative analyses used a priori codes based on an implementation science framework. Quantitative data included demographics, the COVID-19 Impact Scale, and virtual group attendance. Bivariate analyses identified correlates of virtual visit attendance. Results: Eighty percent of mothers reported the pandemic had moderately or extremely impacted at least one major life domain such as daily life, food security, or family conflict. Of 27 mothers offered virtual groups, 67% attended. Mothers who attended virtual groups reported lower English proficiency (p = .087) and fewer friends and family members with COVID-19 (M = 1.0 vs. 5.1, p < .05) than those who did not attend. Women described virtual GWCC as acceptable and a source of social support. Some described differences in group dynamics compared with inperson groups and had privacy concerns. Providers noted scheduling and billing challenges affecting feasibility and sustainability. They reported that visits with good attendance were productive. Mothers and pediatric providers offered recommendations to improve feasibility and privacy and address sustainability. Discussion: Competing demands for those most impacted by COVID-19 may outweigh benefits of attendance. Virtual Spanish language GWCC appears acceptable and feasible for Spanish speaking Latina mothers. Thematic analysis and recommendations identify areas of improvement. Public Significance Statement This study suggests that virtual pediatric group well-child care (GWCC) was generally acceptable and a source of social support for Spanish speaking Latina mothers in the initial phase of the COVID-19 pandemic. Mothers and pediatric providers generated a list of recommendations to improve feasibility, privacy, and address sustainability of GWCC. Keywords: immigrant health, Latino health, pediatric integrated care, telehealth, two-generation approaches to health This article was published Online First June 16. 2022., There is concern that traditional well-child care models do not adequately meet the needs of families experiencing socioeconomic disadvantage (Schor, 2004). Group well-child care (GWCC) is one form of redesign [...]

ABSTRACT: Objectives: The World Health Organization recommends mass drug administration (MDA) with azithromycin to eliminate trachoma as a public health problem. MDA decisions are based on prevalence estimates from two-stage cluster surveys. There is a need to mathematically evaluate current trachoma survey designs. Our study aimed to characterize the effects of the number of units sampled on the precision and cost of trachomatous inflammation–follicular (TF) estimates. Methods: A population of 30 districts was simulated to represent the breadth of possible TF distributions in Amhara, Ethiopia. Samples of varying numbers of clusters (14–34) and households (10–60) were selected. Sampling schemes were evaluated based on precision, proportion of incorrect and low MDA decisions made, and estimated cost. Results: The number of clusters sampled had a greater impact on precision than the number of households. The most efficient scheme depended on the underlying TF prevalence in a district. For lower prevalence areas (< 10%) the most cost-efficient design (providing adequate precision while minimizing cost) was 20 clusters of 20–30 households. For higher prevalence areas (> 10%), the most efficient design was 15–20 clusters of 20–30 households. Conclusions: For longer-running programs, using context-specific survey designs would allow for practical precision while reducing survey costs. Sampling 15 clusters of 20–30 households in suspected moderate-to-high prevalence districts and 20 clusters of 20–30 households in districts suspected to be near the 5% threshold appears to be a balanced approach.

Abstract Membranous nephropathy (MN) is an autoimmune disease caused by autoantibodies against the podocyte antigens phospholipase A2 receptor 1 (PLA2R1) and thrombospondin type 1 domain containing protein 7A (THSD7A) in 80% and 2–3% of patients, respectively. THSD7A antibodies are considered to be pathogenic and highly specific for MN patients. Using an indirect immunofluorescence test (IIFT) we detected THSD7A-antibodies (titre 1:10) in the serum of a patient with high proteinuria who, however, in the kidney biopsy was diagnosed with diabetic nephropathy and MN was excluded as a possible cause of proteinuria. Different immunofluorescence assays and Western blot techniques using recombinant THSD7A (rTHSD7A) or THSD7A from different human tissues revealed that the circulating THSD7A-autoantibodies were only of the IgG3 subclass. The patient serum reacted exclusively with rTHSD7A and only when the antigen was present in reducing Western blot conditions, or on formaldehyde-fixed cells for the IIFT. Our findings show for the first time the existence of circulating THSD7A-antibodies recognizing denatured/reduced rTHSD7A, which do not react with glomerular THSD7A in vivo and are thus presumptively non-pathogenic. As a consequence, kidney biopsy or Western blot analyses of THSD7A under non-reducing conditions should be performed to confirm the diagnosis of THSD7A-associated MN, especially in cases with low THSD7A-antibody levels in the IIFT.

Superfluid vortices are important in many diverse systems, including spinor Bose-Einstein condensates. Here, the experimental and theoretical analysis of the creation and time evolution of vortices in the polar phase of a spin-1 Bose-Einstein condensate is presented, showing the evolution of single-quantum vortices towards half-quantum ones.

Abstract Background There is an emerging body of evidence that links exposure to shift work to cardiovascular disease (CVD). The risk of coronary events, such as myocardial infarction, is greater among night shift workers compared to day workers. There is reason to believe that repeated exposure to shift work, especially night shift work, creates alterations in normal circadian patterns of blood pressure (BP) and heart rate variability (HRV) and that these alterations contribute to increased risk of CVD. Recent data suggest that allowing shift workers to nap during night shifts may help to normalize BP and HRV patterns and, over time, reduce the risk of CVD. The risk of CVD related to shift work is elevated for emergency medical services (EMS) shift workers due in part to long-duration shifts, frequent use of night shifts, and a high prevalence of multiple jobs. Methods We will use a randomized crossover trial study design with three study conditions. The targeted population is comprised of EMS clinician shift workers, and our goal enrollment is 35 total participants with an estimated 10 of the 35 enrolled not completing the study protocol or classified as lost to attrition. All three conditions will involve continuous monitoring over 72 h and will begin with a 36-h at-home period, followed by 24 total hours in the lab (including a 12-h simulated night shift), ending with 12 h at home. The key difference between the three conditions is the intra-shift nap. Condition 1 will involve a simulated 12-h night shift with total sleep deprivation. Condition 2 will involve a simulated 12-h night shift and a 30-min nap opportunity. Condition 3 will involve a simulated 12-h night shift with a 2-h nap opportunity. Our primary outcomes of interest include blunted BP dipping and reduced HRV as measured by the standard deviation of the inter-beat intervals of normal sinus beats. Non-dipping status will be defined as sleep hours BP dip of less than 10%. Discussion Our study will address two indicators of cardiovascular health and determine if shorter or longer duration naps during night shifts have a clinically meaningful impact. Trial registration ClinicalTrials.gov NCT04469803 . Registered on 9 July 2020

Introduction: The World Health Organization (WHO) recommends Transmission Assessment Surveys (TAS) to determine when an evaluation unit (EU) (a designated population survey area) has achieved elimination of transmission of the vector-borne macroparasitic disease Lymphatic Filariasis (LF). These determinations are based on combining data from multiple survey units within an EU; it is unclear how underlying cluster-level variation influences the outcome of the TAS at EU level. We simulate LF infection distribution in an EU and compare three methods for assessing whether LF elimination has occurred based on currently recommended decision thresholds and sampling methods. Methods: We simulate an EU divided into clusters of varying size and disease prevalence. We produce 1000 samples according to LF TAS examples and WHO guidelines and compare three decision-making approaches: lot quality assurance sampling (LQAS) (recommended by WHO), one-sided interval estimate (CI), and nth order statistic (MAX). Summary statistics demonstrating the “pass” rate for the EU under different disease transmission conditions are generated using a versatile SAS® macro. Results: As the prevalence of LF decreases, the LQAS and CI approaches produce increased likelihood of a pass outcome for an EU while some cluster units may still have a high likelihood of transmission. The MAX provides an alternative that increases the likelihood of determining a pass only once the whole area has a low likelihood of transmission. LQAS and CI approaches designed to estimate the LF prevalence in the EU miss hotspots that will continue to transmit infection while the MAX approach focuses on identifying clusters with high risk of transmission. Conclusions: The current TAS methodology has a flaw that may result in false predictions of LF transmission interruption throughout an EU. Modifying the TAS methodology to address results from extreme clusters rather than being based on mean prevalence over an EU will result in greater success for global elimination of LF.

Abstract Background Proprotein convertase subtilisin/kexin type 9 (PCSK9) is of particular importance in cholesterol metabolism with high levels contributing to hypercholesterolemia. Cholesterol and sphingolipids are low in patients with liver cirrhosis. Purpose of this study was to find associations of plasma PCSK9 with circulating cholesterol and sphingolipid species and measures of liver disease severity in patients with liver cirrhosis. Methods PCSK9 protein levels were determined by ELISA in systemic vein (SVP), hepatic vein (HVP) and portal vein plasma of patients with mostly alcoholic liver cirrhosis. PCSK9 and LDL-receptor protein expression were analysed in cirrhotic and non-cirrhotic liver tissues. Results Serum PCSK9 was reduced in patients with liver cirrhosis in comparison to non-cirrhotic patients. In liver cirrhosis, plasma PCSK9 was not correlated with Child-Pugh score, Model for End-Stage Liver Disease score, bilirubin or aminotransferases. A negative association of SVP PCSK9 with albumin existed. PCSK9 protein in the liver did not change with fibrosis stage and was even positively correlated with LDL-receptor protein levels. Ascites volume and variceal size were not related to PCSK9 levels. Along the same line, transjugular intrahepatic shunt to lower portal pressure did not affect PCSK9 concentrations in the three blood compartments. Serum cholesterol, sphingomyelin and ceramide levels did not correlate with PCSK9. Stratifying patients by high versus low PCSK9 levels using the median as cut-off, several cholesteryl ester species were even low in the subgroup with high PCSK9 levels. A few sphingomyelin species were also reduced in the patients with PCSK9 levels above the median. PCSK9 is highly expressed in the liver but systemic, portal and hepatic vein levels were similar. PCSK9 was not correlated with the inflammatory proteins C-reactive protein, IL-6, galectin-3, resistin or pentraxin 3. Of note, HVP PCSK9 was positively associated with HVP chemerin and negatively with HVP adiponectin levels. Conclusions In the cohort of patients with liver cirrhosis mostly secondary to alcohol consumption high PCSK9 was associated with low levels of certain cholesteryl ester and sphingomyelin species. Positive correlations of PCSK9 and LDL-receptor protein in the liver of patients with chronic liver injury are consistent with these findings.

Acinetobacter baumannii is a multidrug-resistant, Gram-negative nosocomial pathogen that exhibits phenotypic heterogeneity resulting in virulent opaque (VIR-O) and avirulent translucent (AV-T) colony variants. Each variant has a distinct gene expression profile resulting in multiple phenotypic differences. Cells interconvert between the VIR-O and AV-T variants at high frequency under laboratory conditions, suggesting that the genetic mechanism underlying the phenotypic switch could be manipulated to attenuate virulence. Therefore, our group has focused on identifying and characterizing genes that regulate this switch, which led to the investigation of ABUW_1132 (1132), a highly conserved gene predicted to encode a LysR-type transcriptional regulator. ABUW_1132 was shown to be a global regulator as the expression of 74 genes was altered ≥ 2-fold in an 1132 deletion mutant. The 1132 deletion also resulted in a 16-fold decrease in VIR-O to AV-T switching, loss of 3-OH-C12-HSL secretion, and reduced surface-associated motility. Further, the deletion of 1132 in the AV-T background caused elevated capsule production, which increased colony opacity and altered the typical avirulent phenotype of translucent cells. These findings distinguish 1132 as a global regulatory gene and advance our understanding of A. baumannii’s opacity-virulence switch.

Summary: Epigenetic modifications (e.g. DNA methylation) in NAFLD and their contribution to disease progression and extrahepatic complications are poorly explored. Here, we use an integrated epigenome and transcriptome analysis of mouse NAFLD hepatocytes and identify alterations in glyoxylate metabolism, a pathway relevant in kidney damage via oxalate release—a harmful waste product and kidney stone-promoting factor. Downregulation and hypermethylation of alanine-glyoxylate aminotransferase (Agxt), which detoxifies glyoxylate, preventing excessive oxalate accumulation, is accompanied by increased oxalate formation after metabolism of the precursor hydroxyproline. Viral-mediated Agxt transfer or inhibiting hydroxyproline catabolism rescues excessive oxalate release. In human steatotic hepatocytes, AGXT is also downregulated and hypermethylated, and in NAFLD adolescents, steatosis severity correlates with urinary oxalate excretion. Thus, this work identifies a reduced capacity of the steatotic liver to detoxify glyoxylate, triggering elevated oxalate, and provides a mechanistic explanation for the increased risk of kidney stones and chronic kidney disease in NAFLD patients.

Introduction:. Reduction mammaplasty is a mainstay in the treatment of symptomatic macromastia, with a well-described positive impact on patient quality-of-life (QoL). Absorbable dermal staplers have the potential to improve the efficiency of skin closure in reduction mammoplasties, but a more comprehensive assessment of its impact on key outcomes has not been fully elucidated. Methods:. A retrospective review of patients undergoing reduction mammoplasty between November 2018 and December 2020 was conducted. Patients were included if they had undergone a wise-pattern reduction with a superomedial pedicle and completed 3 months of follow-up. Patient demographics, operative information, clinical and aesthetic outcomes, and QoL were compared between patients that had INSORB stapler-assisted and suture-only closures. Results:. Seventy-five patients met the inclusion criteria, with 34 patients (45%) in the stapler cohort. Total procedure time was significantly reduced with the use of the dermal stapler (stapler: 154 vs. suture: 170 minutes; p = 0.003). The incidence of major complications was similar between cohorts (stapler: 8.8% vs. suture: 12%; p = 0.64), as was the incidence of minor complications (stapler: 44% vs. suture: 41%; p = 0.82). Regardless of closure technique, patients demonstrated significant increases in all QoL domains (p 0.05). Conclusions:. The dermal stapler improves efficiency of closure during reduction mammoplasty without increasing the incidence of wound healing complications. Additionally, cosmetic outcomes are not affected, and patients demonstrate similar post-operative satisfaction with the result regardless of closure technique.

Climate change, including warmer winter temperatures, a shortened snowfall season, and more rain-on-snow events, threatens nordic skiing as a sport. In response, over-summer snow storage, attempted primarily using woodchips as a cover material, has been successfully employed as a climate change adaptation strategy by high-elevation and/or high-latitude ski centers in Europe and Canada. Such storage has never been attempted at a site that is both low elevation and midlatitude, and few studies have quantified storage losses repeatedly through the summer. Such data, along with tests of different cover strategies, are prerequisites to optimizing snow storage strategies. Here, we assess the rate at which the volume of two woodchip-covered snow piles (each ∼200 m3), emplaced during spring 2018 in Craftsbury, Vermont (45∘ N and 360 m a.s.l.), changed. We used these data to develop an optimized snow storage strategy. In 2019, we tested that strategy on a much larger, 9300 m3 pile. In 2018, we continually logged air-to-snow temperature gradients under different cover layers including rigid foam, open-cell foam, and woodchips both with and without an underlying insulating blanket and an overlying reflective cover. We also measured ground temperatures to a meter depth adjacent to the snow piles and used a snow tube to measure snow density. During both years, we monitored volume change over the melt season using terrestrial laser scanning every 10–14 d from spring to fall. In 2018, snow volume loss ranged from 0.29 to 2.81 m3 d−1, with the highest rates in midsummer and lowest rates in the fall; mean rates of volumetric change were 1.24 and 1.50 m3 d−1, 0.55 % to 0.72 % of initial pile volume per day. Snow density did increase over time, but most volume loss was the result of melting. Wet woodchips underlain by an insulating blanket and covered with a reflective sheet were the most effective cover combination for minimizing melt, likely because the aluminized surface reflected incoming short-wave radiation while the wet woodchips provided significant thermal mass, allowing much of the energy absorbed during the day to be lost by long-wave emission at night. The importance of the pile surface-area-to-volume ratio is demonstrated by 4-fold lower rates of volumetric change for the 9300 m3 pile emplaced in 2019; it lost  % of its initial volume per day between April and October, retaining ∼60 % of the initial snow volume over summer. Together, these data demonstrate the feasibility of over-summer snow storage at midlatitudes and low elevations and suggest efficient cover strategies.

Topological defects and textures are universal phenomena across physics. The authors demonstrate that an initial non-singular spinor texture can be controllably transformed into a pair of singular vortices with cores filled by atoms that continuously connect distinct magnetic phases of matter.

Plastic polymer PET degrading enzymes are of great interest for achieving sustainable plastics recycling. Here, the authors present the crystal structures of the plastic degrading bacterial enzymes PETase, MHETase in its apo-form and MHETase bound to a non-hydrolyzable substrate analog.

Pyruvate (CH3COCOOH) is the simplest of the alpha-keto acids and is at the interface of several metabolic pathways both in prokaryotes and eukaryotes. In an amino acid-rich environment, fast-growing bacteria excrete pyruvate instead of completely metabolizing it. The role of pyruvate uptake in pathological conditions is still unclear. In this study, we identified two pyruvate-specific transporters, BtsT and CstA, in Salmonella enterica serovar Typhimurium (S. Typhimurium). Expression of btsT is induced by the histidine kinase/response regulator system BtsS/BtsR upon sensing extracellular pyruvate, whereas expression of cstA is maximal in the stationary phase. Both pyruvate transporters were found to be important for the uptake of this compound, but also for chemotaxis to pyruvate, survival under oxidative and nitrosative stress, and persistence of S. Typhimurium in response to gentamicin. Compared with the wild-type cells, the ΔbtsTΔcstA mutant has disadvantages in antibiotic persistence in macrophages, as well as in colonization and systemic infection in gnotobiotic mice. These data demonstrate the surprising complexity of the two pyruvate uptake systems in S. Typhimurium.