Bats as instructive animal models for studying longevity and aging.

Bats (order Chiroptera) are emerging as instructive animal models for aging studies. Unlike some common laboratory species, they meet a central criterion for aging studies: they live for a long time in the wild or in captivity, for 20, 30, and even >40 years. Healthy aging (i.e., healthspan) in bats has drawn attention to their potential to improve the lives of aging humans due to bat imperviousness to viral infections, apparent low rate of tumorigenesis, and unique ability to repair DNA. At the same time, bat longevity also permits the accumulation of age-associated systemic pathologies that can be examined in detail and manipulated, especially in captive animals. Research has uncovered additional and critical advantages of bats. In multiple ways, bats are better analogs to humans than are rodents. In this review, we highlight eight diverse areas of bat research with relevance to aging: genome sequencing, telomeres, and DNA repair; immunity and inflammation; hearing; menstruation and menopause; skeletal system and fragility; neurobiology and neurodegeneration; stem cells; and senescence and mortality. These examples demonstrate the broad relevance of the bat as an animal model and point to directions that are particularly important for human aging studies.

Anti-cytomegalovirus antibody levels stratify human immune profiles across the lifespan.

Human cytomegalovirus (hCMV) is a ubiquitous latent persistent herpesvirus infecting 60-90% of the population worldwide. hCMV carriage in immunocompetent people is asymptomatic; thus, hCMV can be considered a component of normative aging. However, hCMV powerfully modulates many features of the immune, and likely other, systems and organs. Questions remain as to how hCMV carriage affects the human host. We used anti-CMV antibody titers as a stratifying criterion to examine the impact of "intensity" of hCMV infection as a potential biomarker of aging, inflammation, and immune homeostasis in a cohort of 247 participants stratified into younger (21-40 years) and older (> 65 years of age) groups. We showed that anti-CMV antibody titers increased with age and directly correlated to increased levels of soluble tumor necrosis factor (sTNFR) I in younger but not older participants. CD8 + cell numbers were reduced in the older group due to the loss in CD8 + T naïve (Tn) cells. In CMV carriers and, in particular, in anti-CMV Ab-high participants, this loss was mitigated or reversed by an increase in the numbers of CD8 + T effector memory (Tem) and T effector memory reexpressing CD45RA (Temra) cells. Analysis of CD38, HLA-DR, and CD57 expression revealed subset (CD4 or CD8)-specific changes that correlated with anti-CMV Ab levels. In addition, anti-CMV Ab levels predicted anti-CMV CD8 T cell responsiveness to different CMV open reading frames (ORFs) selectively in older participants, which correlated to the transcriptional order of expression of specific CMV ORFs. Implications of these results for the potential predictive value of anti-CMV Ab titers during aging are discussed.

Uterine Uptake of Estrogen and Progestogen-Based Radiotracers in Rhesus Macaques with Endometriosis.

PURPOSE: Endometriosis is an estrogen-dependent disorder of menstruating primates where tissues similar to the inner lining of the uterus exist "ectopically" outside of the uterus. The ectopic endometrium, like the endometrium within the uterus, expresses estrogen receptors (ER) and progesterone receptors (PR) and undergoes hormone-dependent cell proliferation and bleeding each menstrual cycle. The goal of this study was to conduct abdominopelvic positron emission tomography (PET) scans with computed tomography (CT) imaging of rhesus macaques (Macaca mulatta) using radiotracers that target ER and PR [16α-[18F]fluoroestradiol (FES) and 12-[18F]fluoro-furanyl-nor-progesterone (FFNP)] in individuals with and without endometriosis. We also aimed to determine if menstrual cycle phase and/or the presence of endometriosis affected the uptake of these radiotracers. PROCEDURES: Rhesus macaques with either clinically diagnosed endometriosis (n = 6) or no endometriosis (n = 4) underwent PET/CT scans with FES. A subset of the animals also underwent PET/CT scans with FFNP. Standard uptake values corrected for body weight (SUVs) were obtained for each radiotracer in target and background tissues (e.g., intestinal). We performed repeated measure analysis of variance tests to determine how uterine and background uptake differed with scan time, phase of the menstrual cycle, and disease state. RESULTS: Abdominopelvic PET/CT could not resolve small, individual endometriotic lesions. However, macaques with endometriosis displayed higher uterine uptake compared to those without the disorder. Radiotracer uptake differed by menstrual cycle phase with increased uterine uptake of both radiotracers in the proliferative phase of the menstrual cycle. Background intestinal uptake of FFNP increased over time after infusion, but only during the proliferative phase. CONCLUSIONS: PET/CT with FES and FFNP support the concept that ER and PR levels are altered in individuals with endometriosis. This highlights the impact of the disease on typical reproductive tract function and may provide a novel pathway for the identification of individuals with endometriosis.

Utilizing Contrast-Enhanced Ultrasonography with Phosphatidylserine Microbubbles to Detect Placental Inflammation in Rhesus Macaques.

The ability to comprehensively monitor physiological and detect pathophysiologic processes early during pregnancy can reduce maternal and fetal morbidity and mortality. Contrast-enhanced ultrasound (CEUS) is a non-invasive imaging technology that utilizes the acoustic detection of microbubbles to examine vascular spaces. Furthermore, microbubbles conjugated to specific compounds can focus studies on precise physiological pathways. We hypothesized that CEUS with phosphatidylserine microbubbles (MB-PS) could be employed to monitor placental inflammation. We tested this hypothesis in rhesus macaques (Macaca mulatta), a translational and relevant animal model of human placental health. As placental inflammation impacts many at-risk pregnancies, we performed CEUS with MB-PS in pregnant macaques fed a high-fat diet (e.g., a western-style diet, WSD) in the presence or absence of testosterone (T) to mimic the increased risk of polycystic ovary syndrome and subfertility. We have previously demonstrated a placental inflammation phenotype in this model, and, thus, we related the MB-PS CEUS signal intensity to placental inflammation markers: selectin p and angiopoietins. Testosterone exposure increased the MB-PS signal in the placental microcirculation on the maternal side compared to control animals. We found that T increased placental weight and decreased angiopoietin 2 (ANGPT2) immunoreactivity. Furthermore, a significant inverse correlation was found between MB-PS signal and ANGPT2. This indicated that CEUS with MB-PS can be used to monitor placental parameters. We propose that CEUS with MB-PS could aid in the identification of pregnancies at risk of placental vascular compromise.

T-cell cellular stress and reticulocyte signatures, but not loss of naïve T lymphocytes, characterize severe COVID-19 in older adults.

In children and younger adults up to 39 years of age, SARS-CoV-2 usually elicits mild symptoms that resemble the common cold. Disease severity increases with age starting at 30 and reaches astounding mortality rates that are ~330 fold higher in persons above 85 years of age compared to those 18-39 years old. To understand age-specific immune pathobiology of COVID-19, we have analyzed soluble mediators, cellular phenotypes, and transcriptome from over 80 COVID-19 patients of varying ages and disease severity, carefully controlling for age as a variable. We found that reticulocyte numbers and peripheral blood transcriptional signatures robustly correlated with disease severity. By contrast, decreased numbers and proportion of naïve T-cells, reported previously as a COVID-19 severity risk factor, were found to be general features of aging and not of COVID-19 severity, as they readily occurred in older participants experiencing only mild or no disease at all. Single-cell transcriptional signatures across age and severity groups showed that severe but not moderate/mild COVID-19 causes cell stress response in different T-cell populations, and some of that stress was unique to old severe participants, suggesting that in severe disease of older adults, these defenders of the organism may be disabled from performing immune protection. These findings shed new light on interactions between age and disease severity in COVID-19.

T-cell cellular stress and reticulocyte signatures, but not loss of naïve T lymphocytes, characterize severe COVID-19 in older adults.

In children and younger adults up to 39 years of age, SARS-CoV-2 usually elicits mild symptoms that resemble the common cold. Disease severity increases with age starting at 30 and reaches astounding mortality rates that are ~330 fold higher in persons above 85 years of age compared to those 18-39 years old. To understand age-specific immune pathobiology of COVID-19 we have analyzed soluble mediators, cellular phenotypes, and transcriptome from over 80 COVID-19 patients of varying ages and disease severity, carefully controlling for age as a variable. We found that reticulocyte numbers and peripheral blood transcriptional signatures robustly correlated with disease severity. By contrast, decreased numbers and proportion of naïve T-cells, reported previously as a COVID-19 severity risk factor, were found to be general features of aging and not of COVID-19 severity, as they readily occurred in older participants experiencing only mild or no disease at all. Single-cell transcriptional signatures across age and severity groups showed that severe but not moderate/mild COVID-19 causes cell stress response in different T-cell populations, and some of that stress was unique to old severe participants, suggesting that in severe disease of older adults, these defenders of the organism may be disabled from performing immune protection. These findings shed new light on interactions between age and disease severity in COVID-19.

Exogenous leptin promotes reproductive behavior during aphagia in red-sided garter snakes (Thamnophis sirtalis parietalis).

Despite the established dichotomy between investment in either reproduction or self-maintenance, a hormonal mechanism that influences an organism's decision to prioritize these behaviors remains elusive. The protein hormone leptin is a likely candidate because it is secreted from adipocytes in proportion to the amount of stored fat in numerous species. Although the majority of studies suggest that leptin stimulates reproduction, the actions of leptin can be context-dependent. Leptin increases sexual behavior in fed individuals, but inhibits sexual behavior in food-restricted individuals. We investigated if exogenous leptin influences sexual behavior in red-sided garter snakes (Thamnophis sirtalis parietalis) experiencing a predictable bout of aphagia during the mating season. We tested two doses of recombinant murine leptin injected for three days. Males were subjected to three mating trials, one on each day of injections, while females were subjected to one mating trial on the last day of injections. Leptin affects male and female snakes similarly by increasing both appetitive (i.e., mating behavior score) and consummatory (i.e., number of copulations, proportion of individuals copulated) sex behavior. We found no evidence to suggest that leptin influenced latency to copulate or duration of copulation. Because leptin promotes reproductive behavior in non-feeding garter snakes, these findings do not align with research on food-restricted mammals. Further investigations into how leptin affects sexual behavior in snakes exposed to food-restriction manipulations would clarify if the role of leptin is evolutionarily divergent.

Energy Metrics of Red-Sided Garter Snakes (Thamnophis sirtalis parietalis) Vary with Sex but Not Life-History Stage.

Because reproduction is energetically expensive, an organism's energy stores are likely involved in mediating transitions between reproductive and self-maintenance activities. We investigated whether body condition index, adipocyte follicle size, and liver glycogen differ with the life-history transition from reproduction to migration and foraging in red-sided garter snakes (Thamnophis sirtalis parietalis). Females primarily investing in mating behavior located at the den had a significantly higher body condition index than females migrating to summer feeding grounds. The body condition index of male snakes did not differ between snakes located at the den and those migrating to summer feeding grounds. Neither adipocyte follicle area nor liver glycogen stores differed significantly between snakes performing mating activities at the den and those migrating to summer feeding grounds. We did find a sexual dimorphism in that female red-sided garter snakes had significantly larger adipocyte follicles and higher liver glycogen compared with males. Our findings support the across-species phenomenon of females and males displaying a sexual dimorphism in stored energy substrates. Conversely, we did not find evidence to suggest that red-sided garter snakes primarily utilize fatty acids to fuel the initiation of migration, a finding that is not consistent with other long-distance migrators, such as birds. Because we did not find evidence to suggest that stored energy metrics influence the decision to migrate, a physiological mechanism that induces migration in red-sided garter snakes remains elusive.

Tidal breathing parameters measured by structured light plethysmography in children aged 2-12 years recovering from acute asthma/wheeze compared with healthy children.

Measurement of lung function can be difficult in young children. Structured light plethysmography (SLP) is a novel, noncontact method of measuring tidal breathing that monitors displacement of the thoraco-abdominal wall. SLP was used to compare breathing in children recovering from an acute exacerbation of asthma/wheeze and an age-matched cohort of controls. Children aged 2-12 years with acute asthma/wheeze (n = 39) underwent two 5-min SLP assessments, one before bronchodilator treatment and one after. SLP was performed once in controls (n = 54). Nonparametric comparisons of patients to healthy children and of pre-bronchodilator to post-bronchodilator were made for all children, and also stratified by age group (2-5 vs. 6-12 years old). In the asthma/wheeze group, IE50SLP (inspiratory to expiratory flow ratio) was higher (median 1.47 vs. 1.31; P = 0.002), thoraco-abdominal asynchrony (TAA) and left-right asynchrony were greater (both P < 0.001), and respiratory rate was faster (P < 0.001) than in controls. All other timing indices were shorter and displayed reduced variability (all P < 0.001). Variability in time to peak inspiratory flow was also reduced (P < 0.001). Younger children showed a greater effect than older children for TAA (interaction P < 0.05). After bronchodilator treatment, the overall cohort showed a reduction in within-subject variability in time to peak expiratory flow only (P < 0.001). Younger children exhibited a reduction in relative contribution of the thorax, TAA, and variability in TAA (interaction P < 0.05). SLP can be successfully performed in young children. The potential of SLP to monitor diseases such as asthma in children is worthy of further investigation. ClinicalTrials.gov identifier: NCT02543333.

Tidal breathing parameters measured using structured light plethysmography in healthy children and those with asthma before and after bronchodilator.

Structured light plethysmography (SLP) is a light-based, noncontact technique that measures tidal breathing by monitoring displacements of the thoracoabdominal (TA) wall. We used SLP to measure tidal breathing parameters and their within-subject variability (v) in 30 children aged 7-16 years with asthma and abnormal spirometry (forced expiratory volume in 1 sec [FEV1] <80% predicted) during a routine clinic appointment. As part of standard care, the reversibility of airway obstruction was assessed by repeating spirometry after administration of an inhaled bronchodilator. In this study, SLP was performed before and after bronchodilator administration, and also once in 41 age-matched controls. In the asthma group, there was a significant increase in spirometry-assessed mean FEV1 after administration of bronchodilator. Of all measured tidal breathing parameters, the most informative was the inspiratory to expiratory TA displacement ratio (IE50SLP, calculated as TIF50SLP/TEF50SLP, where TIF50SLP is tidal inspiratory TA displacement rate at 50% of inspiratory displacement and TEF50SLP is tidal expiratory TA displacement rate at 50% of expiratory displacement). Median (m) IE50SLP and its variability (vIE50SLP) were both higher in children with asthma (prebronchodilator) compared with healthy children (mIE50SLP: 1.53 vs. 1.22, P < 0.001; vIE50SLP: 0.63 vs. 0.47, P < 0.001). After administration of bronchodilators to the asthma group, mIE50SLP decreased from 1.53 to 1.45 (P = 0.01) and vIE50SLP decreased from 0.63 to 0.60 (P = 0.04). SLP-measured tidal breathing parameters could differentiate between children with and without asthma and indicate a response to bronchodilator.

Evaluation of the agreement of tidal breathing parameters measured simultaneously using pneumotachography and structured light plethysmography.

Structured light plethysmography (SLP) is a noncontact, noninvasive, respiratory measurement technique, which uses a structured pattern of light and two cameras to track displacement of the thoraco-abdominal wall during tidal breathing. The primary objective of this study was to examine agreement between tidal breathing parameters measured simultaneously for 45 sec using pneumotachography and SLP in a group of 20 participants with a range of respiratory patterns ("primary cohort"). To examine repeatability of the agreement, an additional 21 healthy subjects ("repeatability cohort") were measured twice during resting breathing and once during increased respiratory rate (RR). Breath-by-breath and averaged RR, inspiratory time (tI), expiratory time (tE), total breath time (tTot), tI/tE, tI/tTot, and IE50 (inspiratory to expiratory flow measured at 50% of tidal volume) were calculated. Bland-Altman plots were used to assess the agreement. In the primary cohort, breath-by-breath agreement for RR was ±1.44 breaths per minute (brpm). tI, tE, and tTot agreed to ±0.22, ±0.29, and ±0.32 sec, respectively, and tI/tE, tI/tTot, and IE50/IE50SLP to ±0.16, ±0.05, and ±0.55, respectively. When averaged, agreement for RR was ±0.19 brpm. tI, tE, and tTot were within ±0.16, ±0.16, and ±0.07 sec, respectively, and tI/tE, tI/tTot, and IE50 were within ±0.09, ±0.03, and ±0.25, respectively. A comparison of resting breathing demonstrated that breath-by-breath and averaged agreements for all seven parameters were repeatable (P > 0.05). With increased RR, agreement improved for tI, tE, and tTot (P ≤ 0.01), did not differ for tI/tE, tI/tTot, and IE50 (P > 0.05) and reduced for breath-by-breath (P < 0.05) but not averaged RR (P > 0.05).

Tidal breathing patterns derived from structured light plethysmography in COPD patients compared with healthy subjects.

PURPOSE: Differences in tidal breathing patterns have been reported between patients with chronic obstructive pulmonary disease (COPD) and healthy individuals using traditional measurement techniques. This feasibility study examined whether structured light plethysmography (SLP) - a noncontact, light-based technique - could also detect differences in tidal breathing patterns between patients with COPD and healthy subjects. PATIENTS AND METHODS: A 5 min period of tidal (quiet) breathing was recorded in each patient with COPD (n=31) and each healthy subject (n=31), matched for age, body mass index, and sex. For every participant, the median and interquartile range (IQR; denoting within-subject variability) of 12 tidal breathing parameters were calculated. Individual data were then combined by cohort and summarized by its median and IQR. RESULTS: After correction for multiple comparisons, inspiratory time (median tI) and its variability (IQR of tI) were lower in patients with COPD (p<0.001 and p<0.01, respectively) as were ratios derived from tI (tI/tE and tI/tTot, both p<0.01) and their variability (p<0.01 and p<0.05, respectively). IE50SLP (the ratio of inspiratory to expiratory flow at 50% tidal volume calculated from the SLP signal) was higher (p<0.001) in COPD while SLP-derived time to reach peak tidal expiratory flow over expiratory time (median tPTEFSLP/tE) was shorter (p<0.01) and considerably less variable (p<0.001). Thoraco-abdominal asynchrony was increased (p<0.05) in COPD. CONCLUSION: These early observations suggest that, like traditional techniques, SLP is able to detect different breathing patterns in COPD patients compared with subjects with no respiratory disease. This provides support for further investigation into the potential uses of SLP in assessing clinical conditions and interventions.

Measuring changes in chest wall motion after lung resection using structured light plethysmography: a feasibility study.

OBJECTIVES: We describe the use of structured light plethysmography (SLP)-a novel, non-contact, light-based technique for measuring tidal breathing-among a cohort of patients undergoing lung resection. In this feasibility study, we examined whether changes in chest wall motion or in asynchrony between regions of the thoraco-abdominal wall could be identified after surgery. METHODS: Fifteen patients underwent wedge resection (n = 8) or lobectomy (n = 7). All patients underwent two SLP assessments (before surgery and on Day 1 post-surgery). Each assessment captured data during 5 min of quiet (tidal) breathing. RESULTS: When data were averaged across all patients, motion on the operated side of the thorax was significantly reduced after surgery (mean change from presurgery ± standard deviation: -14.7 ± 16.5%, P = 0.01), while motion on the non-operated side increased (15.9 ± 18.5%, P = 0.01). Thoraco-abdominal asynchrony also increased (mean change ± standard deviation: 43.4 ± 55.1%, P = 0.01), but no significant difference was observed in right-left hemi-thoracic asynchrony (163.7 ± 230.3%, P = 0.08). When analysed by resection type, lobectomy was associated with reduced and increased motion on the operated and non-operated side, respectively, and with an increase in both right-left hemi-thoracic and thoraco-abdominal asynchrony. No significant changes in motion or asynchrony were identified in patients who underwent wedge resection. CONCLUSIONS: SLP was able to detect changes in chest wall motion and asynchrony after thoracic surgery. Changes in this small group of patients were consistent with the side of the incision and were most apparent in patients undergoing lobectomy.

Diminished size-weight illusion in anorexia nervosa: evidence for visuo-proprioceptive integration deficit.

Individuals with anorexia nervosa (AN) experience pronounced body image distortion in combination with a pernicious desire to maintain a dangerously low body weight. Relatively little is known, however, about the mechanism underlying body image distortion in AN. Despite having normal visual perception, individuals with AN both feel and see themselves as large-bodied and show deficits in interoception and haptic perception, suggesting a potential deficit in visual and tactile integration. The size-weight illusion (SWI) arises when two objects of equal weight but different sizes are held. Typical individuals experience a strong and robust illusion that the smaller object feels much heavier than the larger object because of an implicit assumption that weight scales with size. The current study compared the strength of the SWI in individuals with AN to healthy control participants. Individuals with AN exhibited a markedly reduced SWI relative to controls, even though their ability to discriminate weight was unaffected. Because the SWI is strongly modulated by visual appearance, we believe our finding reflects decreased integration of visual and proprioceptive information in anorexia. This finding may explain the puzzling observation that visual perception of the body in a mirror does not correct an AN patient's distorted body image. We speculate that methods to correct visuo-proprioceptive integration in constructing body image may help rehabilitate patients' judgments of size and weight regarding their own bodies. We also suggest that a dysfunction in interactions between inferior parietal lobule (concerned with body image), insula, and hypothalamus may underlie AN.