Altered ontogeny and transcriptomic signatures of tissue-resident pulmonary interstitial macrophages ameliorate allergic airway hyperresponsiveness.

Introduction: Environmental exposures and experimental manipulations can alter the ontogenetic composition of tissue-resident macrophages. However, the impact of these alterations on subsequent immune responses, particularly in allergic airway diseases, remains poorly understood. This study aims to elucidate the significance of modified macrophage ontogeny resulting from environmental exposures on allergic airway responses to house dust mite (HDM) allergen. Methods: We utilized embryonic lineage labeling to delineate the ontogenetic profile of tissue-resident macrophages at baseline and following the resolution of repeated lipopolysaccharide (LPS)-induced lung injury. We investigated differences in house dust mite (HDM)-induced allergy to assess the influence of macrophage ontogeny on allergic airway responses. Additionally, we employed single-cell RNA sequencing (scRNAseq) and immunofluorescent staining to characterize the pulmonary macrophage composition, associated pathways, and tissue localization. Results: Our findings demonstrate that the ontogeny of homeostatic alveolar and interstitial macrophages is altered after the resolution from repeated LPS-induced lung injury, leading to the replacement of embryonic-derived by bone marrow-derived macrophages. This shift in macrophage ontogeny is associated with reduced HDM-induced allergic airway responses. Through scRNAseq and immunofluorescent staining, we identified a distinct subset of resident-derived interstitial macrophages expressing genes associated with allergic airway diseases, localized adjacent to terminal bronchi, and diminished by prior LPS exposure. Discussion: These results suggest a pivotal role for pulmonary macrophage ontogeny in modulating allergic airway responses. Moreover, our findings highlight the implications of prior environmental exposures in shaping future immune responses and influencing the development of allergies. By elucidating the mechanisms underlying these phenomena, this study provides valuable insights into potential therapeutic targets for allergic airway diseases and avenues for further research into immune modulation and allergic disease prevention.

Tissue-Resident Alveolar Macrophages Reduce Ozone-induced Inflammation via MerTK-mediated Efferocytosis.

Lung inflammation, caused by acute exposure to ozone (O3), one of the six criteria air pollutants, is a significant source of morbidity in susceptible individuals. Alveolar macrophages (AMØs) are the most abundant immune cells in the normal lung, and their number increases after O3 exposure. However, the role of AMØs in promoting or limiting O3-induced lung inflammation has not been clearly defined. In this study, we used a mouse model of acute O3 exposure, lineage tracing, genetic knockouts, and data from O3-exposed human volunteers to define the role and ontogeny of AMØs during acute O3 exposure. Lineage-tracing experiments showed that 12, 24, and 72 hours after exposure to O3 (2 ppm) for 3 hours, all AMØs were of tissue-resident origin. Similarly, in humans exposed to filtered air and O3 (200 ppb) for 135 minutes, we did not observe at ∼21 hours postexposure an increase in monocyte-derived AMØs by flow cytometry. Highlighting a role for tissue-resident AMØs, we demonstrate that depletion of tissue-resident AMØs with clodronate-loaded liposomes led to persistence of neutrophils in the alveolar space after O3 exposure, suggesting that impaired neutrophil clearance (i.e., efferocytosis) leads to prolonged lung inflammation. Moreover, depletion of tissue-resident AMØs demonstrated reduced clearance of intratracheally instilled apoptotic Jurkat cells, consistent with reduced efferocytosis. Genetic ablation of MerTK (MER proto-oncogene, tyrosine kinase), a key receptor involved in efferocytosis, also resulted in impaired clearance of apoptotic neutrophils after O3 exposure. Overall, these findings underscore the pivotal role of tissue-resident AMØs in resolving O3-induced inflammation via MerTK-mediated efferocytosis.

Tissue-resident alveolar macrophages reduce O3-induced inflammation via MerTK mediated efferocytosis.

Lung inflammation, caused by acute exposure to ozone (O3) - one of the six criteria air pollutants - is a significant source of morbidity in susceptible individuals. Alveolar macrophages (AMØs) are the most abundant immune cells in the normal lung and their number increases following O3 exposure. However, the role of AMØs in promoting or limiting O3-induced lung inflammation has not been clearly defined. Here, we used a mouse model of acute O3 exposure, lineage tracing, genetic knockouts, and data from O3-exposed human volunteers to define the role and ontogeny of AMØs during acute O3 exposure. Lineage tracing experiments showed that 12, 24, and 72 h after exposure to O3 (2 ppm) for 3h all AMØs were tissue-resident origin. Similarly, in humans exposed to FA and O3 (200 ppb) for 135 minutes, we did not observe ~21h post-exposure an increase in monocyte-derived AMØs by flow cytometry. Highlighting a role for tissue-resident AMØs, we demonstrate that depletion of tissue-resident AMØs with clodronate-loaded liposomes led to persistence of neutrophils in the alveolar space after O3 exposure, suggesting that impaired neutrophil clearance (i.e., efferocytosis) leads to prolonged lung inflammation. Moreover, depletion of tissue-resident AMØ demonstrated reduced clearance of intratracheally instilled apoptotic Jurkat cells, consistent with reduced efferocytosis. Genetic ablation of MerTK - a key receptor involved in efferocytosis - also resulted in impaired clearance of apoptotic neutrophils followed O3 exposure. Overall, these findings underscore the pivotal role of tissue-resident AMØs in resolving O3-induced inflammation via MerTK-mediated efferocytosis.

Allergic Asthma Responses Are Dependent on Macrophage Ontogeny.

The ontogenetic composition of tissue-resident macrophages following injury, environmental exposure, or experimental depletion can be altered upon re-establishment of homeostasis. However, the impact of altered resident macrophage ontogenetic milieu on subsequent immune responses is poorly understood. Hence, we assessed the effect of macrophage ontogeny alteration following return to homeostasis on subsequent allergic airway responses to house dust mites (HDM). Using lineage tracing, we confirmed alveolar and interstitial macrophage ontogeny and their replacement by bone marrow-derived macrophages following LPS exposure. This alteration in macrophage ontogenetic milieu reduced allergic airway responses to HDM challenge. In addition, we defined a distinct population of resident-derived interstitial macrophages expressing allergic airway disease genes, located adjacent to terminal bronchi, and reduced by prior LPS exposure. These findings support that the ontogenetic milieu of pulmonary macrophages is a central factor in allergic airway responses and has implications for how prior environmental exposures impact subsequent immune responses and the development of allergy.

Frailty and Cause-Specific Hospitalizations in Community-Dwelling Older Men.

OBJECTIVES: The types of medical conditions leading to hospitalization in frail older people have not been investigated. The objectives were to evaluate associations between frailty and (a) risk of all-cause and cause-specific hospitalization, and (b) rate of all-cause and cause-specific hospitalizations. DESIGN, SETTING AND PARTICIPANTS: Community-dwelling men aged 70+ years in the Concord Health and Ageing in Men Project (CHAMP) were assessed for frailty at baseline (2005-2007, n=1705). MEASUREMENTS: Frailty was determined by both the Fried frailty phenotype (FP) and the Rockwood frailty index (FI). Non-elective and elective hospitalization data were accessed from the New South Wales (NSW) Admitted Patient Data Collection and mortality from the NSW Deaths Registry for the period 2005-2017. Causes of hospitalization were categorized using ICD-10 classification of principal diagnoses based on organ system involved into 14 major categories. RESULTS: Nearly 80% of CHAMP men had at least one non-elective hospitalization and 63% had an elective hospitalization over a 9-year follow-up. Men with FP frailty were twice as likely to have a non-elective hospitalization (HR: 1.98, 95%CI: 1.61-2.44) and a greater number of non-elective hospitalizations (IRR: 1.44, 95%CI: 1.22-1.70). Similar relationships were found between FI frailty and non-elective hospitalizations. Men with frailty (either FP or FI) were more likely to have at least one non-elective hospitalization for 13 of the 14 cause-related admissions. In contrast, frailty was only associated with 3 cause-related elective hospitalizations. Men with frailty were also more likely to have an increased number of non-elective hospitalizations for all 14 causes, but only for 6 causes of elective hospitalizations. CONCLUSIONS: Our findings suggest frailty increases the risk and number of non-elective hospitalizations in older men for a wide range of cause. Strategies on early identification of frailty, followed by appropriate preventative strategies to lower the risk of non-elective hospital admissions are warranted.

Nonclassical Monocytes Sense Hypoxia, Regulate Pulmonary Vascular Remodeling, and Promote Pulmonary Hypertension.

An increasing body of evidence suggests that bone marrow-derived myeloid cells play a critical role in the pathophysiology of pulmonary hypertension (PH). However, the true requirement for myeloid cells in PH development has not been demonstrated, and a specific disease-promoting myeloid cell population has not been identified. Using bone marrow chimeras, lineage labeling, and proliferation studies, we determined that, in murine hypoxia-induced PH, Ly6Clo nonclassical monocytes are recruited to small pulmonary arteries and differentiate into pulmonary interstitial macrophages. Accumulation of these nonclassical monocyte-derived pulmonary interstitial macrophages around pulmonary vasculature is associated with increased muscularization of small pulmonary arteries and disease severity. To determine if the sensing of hypoxia by nonclassical monocytes contributes to the development of PH, mice lacking expression of hypoxia-inducible factor-1α in the Ly6Clo monocyte lineage were exposed to hypoxia. In these mice, vascular remodeling and PH severity were significantly reduced. Transcriptome analyses suggest that the Ly6Clo monocyte lineage regulates PH through complement, phagocytosis, Ag presentation, and chemokine/cytokine pathways. Consistent with these murine findings, relative to controls, lungs from pulmonary arterial hypertension patients displayed a significant increase in the frequency of nonclassical monocytes. Taken together, these findings show that, in response to hypoxia, nonclassical monocytes in the lung sense hypoxia, infiltrate small pulmonary arteries, and promote vascular remodeling and development of PH. Our results demonstrate that myeloid cells, specifically cells of the nonclassical monocyte lineage, play a direct role in the pathogenesis of PH.

Circulating Sex Steroid Measurements of Men by Mass Spectrometry Are Highly Reproducible after Prolonged Frozen Storage.

Long-term studies investigating hormone-dependent cancers and reproductive health often require prolonged frozen storage of serum which assumes that the steroid molecules and measurements are stable over that time. Previous studies of reproducibility of circulating steroids have relied upon flawed historical rather than contemporaneous controls. We measured serum testosterone (T), dihydrotestosterone (DHT), estradiol (E2) and estrone (E1) in 150 randomly selected serum samples by liquid chromatography-mass spectrometry (LC-MS) from men 70 years or older (mean age 77 years) in the CHAMP study. The original measurements in 2009 were repeated 10 years later using the identical serum aliquot (having undergone 2-4 freeze-thaw cycles in the interim) in 2019 together with another never-thawed aliquot of the same serum sample. The results of all three sets of measurements were evaluated by Passing-Bablok regression and Bland-Altman difference analysis. Serum androgens (T, DHT) and estrogens (E2, E1) measured by LC-MS display excellent reproducibility when stored for 10 years at -80 C without thawing. Serum T and DHT displayed high level of reproducibility across all three sets of measurements. Multiple freeze-thaw cycles over those storage conditions do not significantly affect serum T, DHT and E1 concentrations but produce a modest increase (21%) in serum E2 measurements.

Immunisation Status of Children with Cerebral Palsy in Rural Bangladesh: Results from the Bangladesh Cerebral Palsy Register (BCPR).

BACKGROUND: Cerebral palsy (CP) is the most common cause of physical disability in childhood, with an estimated 17 million cases worldwide. There is limited data concerning the general health of this population and the immunisation status of children with CP is largely unknown. OBJECTIVE: We aimed to assess the immunisation status of children with CP in rural Bangladesh and determine the predictors of non-immunisation. METHODS: This study is part of the Bangladesh CP Register (BCPR) study; a population based CP register commenced in January 2015 in the Shahjadpur sub-district of Bangladesh. As part of BCPR registration, all children with CP in the catchment area were assessed by a paediatrician and their clinical and immunisation history were collected. RESULTS: Between January and December 2015, 615 children with CP were registered on the BCPR. The median age of the children was 7.5 years, and 38.5% were female. 91.7% of those children had a BCG vaccine scar (as an objective marker for immunisation at birth). However, only 43.2% reported to have received the rubella vaccine during the 2014 national rubella immunisation campaign. Timing of CP diagnosis was found to be an independent predictor for immunisation uptake; those diagnosed before the age of 3 were more likely to have received the rubella vaccine (95% confidence interval [CI] 1.6 - 4.3, odds ratio [OR] 2.6, p <0.0001). CONCLUSIONS: To the best of our knowledge, this is the first paper to use a formal CP register to examine the relationship between CP and immunisation status in a low or middle income country like Bangladesh. Our data suggest that more than half of children with CP in rural Bangladesh did not receive immunisation during a recent national campaign.

Oral health behaviours of older Australian men: the Concord Health and Ageing in Men Project.

BACKGROUND: The Concord Health and Ageing in Men Project (CHAMP) is a cohort study of the health of a representative sample of older Australian men. The aim of this paper is to describe the oral health behaviours and dental service use of CHAMP participants and explore associations between oral health behaviours with and general health status. METHOD: Information collected related to socio-demographics, general health, oral health service-use and oral health behaviours. Key general health conditions were ascertained from the health questionnaire and included physical capacity and cognitive status. RESULTS: Fifty-seven percent of the men reported visiting a dental provider at least once or more a year and 56.7% did so for a "dental check-up". Of those with some natural teeth, 59.3% claimed to brush their teeth at least twice or more a day. Most men (96%) used a standard fluoride toothpaste. Few participants used dental floss, tooth picks or mouth-rinses to supplement oral hygiene. Cognitive status and self-rated general health were associated with dental visiting patterns and toothbrushing behaviour. CONCLUSIONS: Most older men in CHAMP perform favourable oral health behaviours. Smoking behaviour is associated with less favourable dental visiting patterns, and cognitive status with toothbrushing behaviour.

Phenotyping Tumor-Associated Macrophages.

Multi-parametric flow cytometry of tumor-bearing murine nonlymphoid tissue allows for characterization, isolation, and examination of immune cell composition, phenotype, and function. Here we describe an approach to process nonlymphoid tissues and then utilize a base antibody panel to define all of the major immune cell types in a single staining condition. This panel can be used to phenotype tumor-associated macrophages.

Metabolic plasticity during transition to naïve-like pluripotency in canine embryo-derived stem cells.

Pluripotent stem cells (PSCs) have been described in naïve or primed pluripotent states. Domestic dogs are useful translational models in regenerative medicine, but their embryonic stem cells (cESCs) remain narrowly investigated. Primed-like cESCs expanded in the presence of leukemia inhibitory factor and fibroblast growth factor 2 (LIF-FGF2) acquire features of naïve pluripotency when exposed to chemical inhibitors and LIF (2iL). However, proliferation of cESCs is influenced by the pluripotent state and is comparatively slower than human or mouse PSCs. We propose that different metabolic pathway activities support ATP generation and biomass accumulation necessary for LIF-FGF2 and 2iL cESC proliferation. We found that 2iL cESCs have greater respiratory capacity, altered mitochondrial chain complex stoichiometry and elevated mitochondrial polarization state. Yet, 2iL-enriched cESCs exhibited immature ultrastructure, including previously unrecognized changes to cristae organization. Enhanced ATP level in 2iL cESCs is associated with altered retrograde signalling, whereas LIF-FGF2 cESCs exhibit a lipogenic phenotype. Inhibition of oxidative phosphorylation impaired proliferation and ATP production in 2iL cESCs but not LIF-FGF2 cESCs, which remained sensitive to glycolysis inhibition. Our study reveals distinct bioenergetic mechanisms contributing to steady-state expansion of distinct canine pluripotent states that can be exploited to improve derivation and culture of canine PSCs.

Oral health of community-dwelling older Australian men: the Concord Health and Ageing in Men Project (CHAMP).

BACKGROUND: The Concord Health and Ageing in Men Project (CHAMP) is a cohort study of the health of a representative sample of Australian men aged 70 years and older. The aim of this report is to describe the oral health of these men. METHODS: Oral health was assessed when the men were all aged 78 years or older. Two calibrated examiners conducted a standardized intraoral assessment. Descriptive data were analysed by statistical association tests. Participants were excluded from the collection of some periodontal assessments if they had a medical contraindication. RESULTS: Dental assessments of 614 participants revealed 90 (14.6%) were edentate. Men had a mean of 13.8 missing teeth and 10.3 filled teeth. Dentate participants had a mean of 1.1 teeth with active coronal decay. Those in the low-income group had a higher rate of decayed teeth and lower rate of filled teeth. Thirty-four participants (5.5%) had one or more dental implants, and 66.3% relied on substitute natural teeth for functional occlusion. Of those with full periodontal assessments; 90.9% had sites with pocket depths of 3 mm or more, 96.6% had sites with CAL of 5 mm or more, and 79.7% had three or more sites with GI scores of 2 or more. CONCLUSIONS: There was a high prevalence of periodontal diseases and restorative burden of dentitions, which suggests that greater attention needs to be given to prevention and health maintenance in older Australian men.

Health and social support services in older adults recently discharged from hospital: service utilisation and costs and exploration of the impact of a home-exercise intervention.

BACKGROUND: Admission to hospital can lead to persistent deterioration in physical functioning, particularly for the more vulnerable older population. As a result of this physical deterioration, older people who have been recently discharged from hospital may be particularly high users of health and social support services. Quantify usage and costs of services in older adults after hospitalisation and explore the impact of a home-exercise intervention on service usage. METHOD: The present study was a secondary analysis of data from a randomised controlled trial (ACTRN12607000563460). The trial involved 340 participants aged 60 years and over with recent hospitalisation. Service use and costs were compared between intervention (12 months of home-exercise prescribed in 10 visits from a physiotherapist) and control groups. RESULTS: 33 % of participants were re-admitted to hospital, 100 % consulted a General Medical Practitioner and 63 % used social services. 56 % of costs were associated with hospital admission and 22 % with social services. There was reduction in General Medical Practitioner services provided in the home in the intervention group (IRR 0.23, CI 0.1 to 0.545, p < 0.01) but no significant between-group difference in service use or in costs for other service categories. CONCLUSION: There appears to be substantial hospital and social service use and costs in this population of older people. No significant impact of a home-based exercise program was evident on service use or costs. TRIAL REGISTRATION: Australian and New Zealand Clinical Trial Registry ACTRN12607000563460 >TrialSearch.

Atrial fibrillation in older inpatients: are there any differences in clinical characteristics and pharmacological treatment between the frail and the non-frail?

BACKGROUND: Frailty is common in patients with atrial fibrillation and may impact on antithrombotic and anti-arrhythmic treatment. AIM: To describe differences in clinical characteristics, prescription of antithrombotic and anti-arrhythmic medications and incidence of haemorrhage and stroke, between frail and non-frail older inpatients. METHODS: Prospective observational study in patients aged ≥65 years with atrial fibrillation admitted to a teaching hospital in Sydney, Australia. Frailty was assessed using the Reported Edmonton Frail Scale, stroke risk with CHA2DS2-VASc score and bleeding risk with HAS-BLED score. Participants were followed after 6 months for haemorrhages and strokes. RESULTS: We recruited 302 patients (mean age 84.7 ± 7.1 years, 53.3% frail, 50% female, mean CHA2DS2-VASc 4.61 ± 1.44, mean HAS-BLED 2.97 ± 1.04). Frail participants were older and had more co-morbidities and higher risk of stroke but not haemorrhage. Upon discharge, 55.7% participants were prescribed with anticoagulants (49.3% frail, 62.6% non-frail, P = 0.02). Thirty-three per cent received antiplatelets only and 11.1% no antithrombotics, with no difference by frailty status. For anti-arrhythmics, 52.6% received rate-control drugs only, 11.8% rhythm-control drugs only and 13.5% both and 22.1% were not prescribed either, with no difference by frailty status. On univariate logistic regression, frailty decreased the likelihood of anticoagulant prescription (odds ratio (OR) 0.58, 95%CI 0.36-0.93), but this was not significant on multivariate analysis (OR 0.66, 95%CI 0.40-1.11). After 6 months, overall incidence of ischaemic stroke was 2.1%, and in patients taking anticoagulants, incidence of major/severe bleeding was 6.3%, with no significant difference between frailty groups. CONCLUSIONS: Frailty status had little impact on antithrombotic prescription and no impact on anti-arrhythmic prescription.

A Review of Frailty in Developing Countries.

BACKGROUND: As the population ages, the prevalence and clinical importance of frailty are increasing. There have been few published studies about frailty in developing world. This study aims to review the evidence from developing countries on the prevalence of frailty, definition of frailty and factors associated with frailty. METHOD: A literature search was conducted via MEDLINE and EMBASE. Keywords included "frail", "frailty", "prevalence", "criteria", "definition", "risk factors", "outcomes", "developing country", "developing world", and names of low and middle income countries according to the classification of the World Bank. RESULT: A total of 14 articles were reviewed from Brazil (n=6), China (n=3), Mexico (n=2), and one each from Russia, India, and Peru. There were 9 articles from community-based studies and 5 articles from hospital-based studies. Fried's phenotype for frailty was used to define frailty in the majority of studies. The prevalence of frailty in community-dwelling older people was 17%-31% in Brazil, 15% in Mexico, 5%-31% in China, and 21%-44% in Russia. The prevalence of frailty was 49% in institutionalized older patients in Brazil and 32% in hospitalized older patients in India. The prevalence of frailty in outpatient clinics was 55%-71% in Brazil and 28% in Peru. Frailty was associated with increased mortality and comorbidities, decreased physical and cognitive function, and poor perceptions of health. CONCLUSION: The limited studies available suggest that frailty occurs frequently in older people in the developing world and it appears to be associated with adverse outcomes. This has implications for policy and health care provision for these ageing populations.

Relative Validity of a Diet History Questionnaire Against a Four-Day Weighed Food Record among Older Men in Australia: The Concord Health and Ageing in Men Project (CHAMP).

OBJECTIVES: To evaluate the relative validity of the diet history questionnaire (DHQ) used in the Concord Health and Ageing in Men Project (CHAMP) against a four-day weighed food record (4dWFR) as the reference method. DESIGN AND MEASUREMENTS: Detailed DHQ followed by a 4dWFR were completed between July 2012 and October of 2013. SETTING: Burwood, Canada Bay and Strathfield in Sydney, Australia. PARTICIPANTS: Fifty six community- dwelling men aged 75 years and over (mean=79 years). RESULTS: DHQ estimates of intakes were generally higher than estimates from 4dWFR. Differences between the two methods were generally less than 20% with the exception of ß-carotene (37%). Fixed and proportional biases were only present for retinol, ß-carotene, magnesium, phosphorus and percentage of energy from protein; however, 95% limits of agreement were in some cases wide. Pearson correlation coefficient of log-transformed unadjusted values ranged from 0.15 (zinc) to 0.70 (alcohol), and from 0.06 (iron) to 0.63 (thiamin) after energy-adjustment. Spearman's correlation coefficients ranged from 0.16 (zinc) to 0.80 (alcohol) before energy adjustment, and from 0.15(zinc) to 0.81(alcohol) after energy adjustment. CONCLUSION: Our findings suggest that the DHQ used in CHAMP to measure the nutritional intake of its participants is appropriate to this age group and provides reasonably similar results to the 4dWFR for the majority of nutrients analysed.

A new genus of Stenetriidae Hansen, 1905 (Asellota: Isopoda: Crustacea) from the Great Barrier Reef, Australia and the southwestern Pacific.

Onychatrium gen. nov. is described, with five included species: Onychatrium forceps sp. nov., the type species and Onychatrium torosus sp. nov., both from the Great Barrier Reef; Onychatrium entale (Nordenstam, 1946) comb. nov., from Tapateuen (= Tabiteue Island), Gilbert Islands; Onychatrium thomasi (Bolstad & Kensley, 1999) comb. nov., from Madang, Papua New Guinea; and Onychatrium echiurum (Nobili, 1906) comb. nov., and species inquirenda from the Tumaotu Islands, Eastern French Polynesia. The primary distinguishing characters for Onychatrium gen. nov. are a trapezoid pseudosrostrum, the male pereopod 1 with elongate dactylus (4.7-7.3 as long as proximal width), propodus with strongly produced and acute lobe, carpus with a distally acute, flat, ventrally directed process (except O. torosus sp. nov., which has a short and truncate process) and the merus with a distally directed inferodistal lobe. The genus is known only from the southern Pacific, from the Tuamotus (eastern French Polynesia) to the Great Barrier Reef and northern Papua New Guinea.

Statin utilisation patterns in older Australians living in residential care: 1-year prevalence study.

Despite controversy over the risks and benefits of statin therapy, statins continue to be commonly used medicines by older people. In a cohort study of participants aged ≥70 years (n = 540) living in residential care, Sydney, we found that the proportion of statin users decreased gradually from the baseline of 33.1% to 31.3% at 6 months (P = 0.13) and to 28.7% over 1 year (P = 0.002). Prevalence of statin use decreased with increasing age, with individuals aged ≥90 years being more likely to discontinue or deprescribe statins. The patterns of statin use did not change according to increasing baseline dose or baseline indication.

Identification of early interactions between Francisella and the host.

The adaptive immune response to Francisella tularensis is dependent on the route of inoculation. Intradermal inoculation with the F. tularensis live vaccine strain (LVS) results in a robust Th1 response in the lungs, whereas intranasal inoculation produces fewer Th1 cells and instead many Th17 cells. Interestingly, bacterial loads in the lungs are similar early after inoculation by these two routes. We hypothesize that the adaptive immune response is influenced by local events in the lungs, such as the type of cells that are first infected with Francisella. Using fluorescence-activated cell sorting, we identified alveolar macrophages as the first cell type infected in the lungs of mice intranasally inoculated with F. novicida U112, LVS, or F. tularensis Schu S4. Following bacterial dissemination from the skin to the lung, interstitial macrophages or neutrophils are infected. Overall, we identified the early interactions between Francisella and the host following two different routes of inoculation.