Senescent hearts from male Ts65Dn mice exhibit preserved function but altered size and nicotinamide adenine dinucleotide pathway signaling.

Down syndrome (DS) is associated with congenital heart defects at birth, but cardiac function has not been assessed at older ages. We used the Ts65Dn mouse, a model of DS, to quantify heart structure and function with echocardiography in 18-mo male Ts65Dn and wild-type (WT) mice. Heart weight, nicotinamide adenine dinucleotide (NAD) signaling, and mitochondrial (citrate synthase) activity were investigated, as these pathways may be implicated in the cardiac pathology of DS. The left ventricle was smaller in Ts65Dn versus WT, as well as the anterior wall thickness of the left ventricle during both diastole (LVAW_d; mm) and systole (LVAW_s; mm) as assessed by echocardiography. Other functional metrics were similar between groups including left ventricular area end systole (mm2), left ventricular area end diastole (mm2), left ventricular diameter end systole (mm), left ventricular diameter end diastole (mm), isovolumetric relaxation time (ms), mitral valve atrial peak velocity (mm/s), mitral valve early peak velocity (mm/s), ratio of atrial and early peak velocities (E/A), heart rate (beats/min), ejection fraction (%), and fractional shortening (%). Nicotinamide phosphoribosyltransferase (NAMPT) protein expression, NAD concentration, and tissue weight were lower in the left ventricle of Ts65Dn versus WT mice. Sirtuin 3 (SIRT3) protein expression and citrate synthase activity were not different between groups. Although cardiac function was generally preserved in male Ts65Dn, the altered heart size and bioenergetic disturbances may contribute to differences in aging for DS.

Breathing and Oxygen Carrying Capacity in Ts65Dn and Down Syndrome.

Individuals with Down syndrome (Ds) are at increased risk of respiratory infection, aspiration pneumonia, and apnea. The Ts65Dn mouse is a commonly used model of Ds, but there have been no formal investigations of awake breathing and respiratory muscle function in these mice. We hypothesized that breathing would be impaired in Ts65Dn vs. wild-type (WT), and would be mediated by both neural and muscular inputs. Baseline minute ventilation was not different at 3, 6, or 12 mo of age. However, VT/Ti, a marker of the neural drive to breathe, was lower in Ts65Dn vs. WT and central apneas were more prevalent. The response to breathing hypoxia was not different, but the response to hypercapnia was attenuated, revealing a difference in carbon dioxide sensing, and/or motor output in Ts65Dn. Oxygen desaturations were present in room air, demonstrating that ventilation may not be sufficient to maintain adequate oxygen saturation in Ts65Dn. We observed no differences in arterial PO2 or PCO2, but Ts65Dn had lower hemoglobin and hematocrit. A retrospective medical record review of 52,346 Ds and 52,346 controls confirmed an elevated relative risk of anemia in Ds. We also performed eupneic in-vivo electromyography and in-vitro muscle function and histological fiber typing of the diaphragm, and found no difference between strains. Overall, conscious respiration is impaired in Ts65Dn, is mediated by neural mechanisms, and results in reduced hemoglobin saturation. Oxygen carrying capacity is reduced in Ts65Dn vs. WT, and we demonstrate that individuals with Ds are also at increased risk of anemia.

Is That "Mr." or "Ms." Lemon? An Investigation of Grammatical and Semantic Gender on the Perception of Household Odorants.

Linguistic relativism is the idea that the structure of language influences thought. The present study investigates linguistic relativism by asking whether people who speak a gendered language think of objects in a way that is consistent with the grammatical gender (more masculine or feminine) more than people who speak a language without that linguistic feature. Odorants are unique stimuli for examining this question, because they carry a semantic gender without a biological one, yet gender is thought to be a central dimension of their perception. Canadian participants in the present study (native French or native English speakers) described a set of labeled odorants that varied systematically in grammatical and semantic gender to produce an implicit gender measure and also explicitly rated them for gender. Analysis of the implicit femininity contained in participants' descriptions showed a tendency for both native French and native English speakers to produce descriptions that were consistent with semantic gender; moreover, there were no effects of grammatical gender on implicit or explicit ratings. These results suggest that in a bilingual culture, French speakers may have been influenced by the anthropomorphism associated with odorants rather than grammatical gender.

Method to Obtain Pattern of Breathing in Senescent Mice through Unrestrained Barometric Plethysmography.

Unrestrained barometric plethysmography (UBP) is a method for quantifying the pattern of breathing in mice, where breathing frequency, tidal volume, and minute ventilation are routinely reported. Moreover, information can be collected regarding the neural output of breathing, including the existence of central apneas and augmented breaths. An important consideration for UBP is obtaining a breathing segment with a minimal impact of anxious or active behaviors, to elucidate the response to breathing challenges. Here, we present a protocol that allows for short, quiet baselines to be obtained in aged mice, comparable to waiting for longer bouts of quiet breathing. The use of shorter time segments is valuable, as some strains of mice may be increasingly excitable or anxious, and longer periods of quiet breathing may not be achieved within a reasonable timeframe. We placed 22 month-old mice in a UBP chamber and compared four 15 s quiet breathing segments between minutes 60-120 to a longer 10 min quiet breathing period that took 2-3 h to acquire. We also obtained counts of central apneas and augmented breaths prior to the quiet breathing segments, following a 30 min familiarization period. We show that 10 min of quiet breathing is comparable to using a much shorter 15 s duration. Additionally, the time leading up to these 15 s quiet breathing segments can be used to gather data regarding apneas of central origin. This protocol allows investigators to collect pattern-of-breathing data in a set amount of time and makes quiet baseline measures feasible for mice that may exhibit increased amounts of excitable behavior. The UBP methodology itself provides a useful and noninvasive way to collect pattern-of-breathing data and allows for mice to be tested over several time points.

Heart rate and blood pressure in male Ts65Dn mice: a model to investigate cardiovascular responses in Down syndrome.

Down syndrome (Ds) is the most common chromosomal cause of intellectual disability that results from triplication of chromosome 21 genes. Lower blood pressure (BP) and heart rate (HR) in response to exercise and other stressors are prevalent in Ds, and are mediated by autonomic dysfunction. The Ts65Dn mouse is a model of Ds that is commonly used in preclinical studies, but has not been formally investigated for cardiovascular responses in conscious mice. Based on human studies of Ds, we hypothesized Ts65Dn would have lower BP and HR, but similar arterial stiffness. BP was quantified in conscious wild-type (WT) and Ts65Dn. A main effect for strain was observed for all BP measures (systolic, diastolic, mean, pulse pressure), with WT higher than Ts65Dn. Pulse wave velocity was similar between WT and Ts65Dn. High-frequency power spectra was higher in WT suggesting autonomic differences between strains. Freely moving HR was higher in WT versus Ts65Dn in both the dark and light cycles, although a main effect of circadian cycle was also present (dark> light). Similar to what is observed in humans, Ts65Dn has a lower BP which may be attributed to autonomic differences and result in preservation of arterial function with advancing age. Ts65Dn thus appears to capture the Ds cardiovascular phenotype across the lifespan. These data support further use of Ts65Dn to investigate mechanisms that may lead to altered BP and HR responses in Ds.

Young and middle-aged mouse breathing behavior during the light and dark cycles.

Unrestrained barometric plethysmography is a common method used for characterizing breathing patterns in small animals. One source of variation between unrestrained barometric plethysmography studies is the segment of baseline. Baseline may be analyzed as a predetermined time-point, or using tailored segments when each animal is visually calm. We compared a quiet, minimally active (no sniffing/grooming) breathing segment to a predetermined time-point at 1 h for baseline measurements in young and middle-aged mice during the dark and light cycles. Additionally, we evaluated the magnitude of change for gas challenges based on these two baseline segments. C57BL/6JEiJ x C3Sn.BliA-Pde6b+ /DnJ male mice underwent unrestrained barometric plethysmography with the following baselines used to determine breathing frequency, tidal volume (VT) and minute ventilation (VE): (1) 30-sec of quiet breathing and (2) a 10-min period from 50 to 60 min. Animals were also exposed to 10 min of hypoxic (10% O2 , balanced N2 ), hypercapnic (5% CO2 , balanced air) and hypoxic hypercapnic (10% O2 , 5% CO2 , balanced N2 ) gas. Both frequency and VE were higher during the predetermined 10-min baseline versus the 30-sec baseline, while VT was lower (P < 0.05). However, VE/VO2 was similar between the baseline time segments (P > 0.05) in an analysis of one cohort. During baseline, dark cycle testing had increased VT values versus those in the light (P < 0.05). For gas challenges, both frequency and VE showed higher percent change from the 30-sec baseline compared to the predetermined 10-min baseline (P < 0.05), while VT showed a greater change from the 10-min baseline (P < 0.05). Dark cycle hypoxic exposure resulted in larger percent change in breathing frequency versus the light cycle (P < 0.05). Overall, light and dark cycle pattern of breathing differences emerged along with differences between the 30-sec behavior observational method versus a predetermined time segment for baseline.

Minute ventilation during hypoxia is augmented with capsaicin supplementation in aged mice.

Capsaicin is an agonist for transient receptor potential vanilloid 1 (TRPV1), and acute injection results in an increased frequency and tidal volume in young rats. It is unknown how capsaicin influences breathing in aged mice. We tested the hypothesis that capsaicin supplementation would elicit an augmented pattern of breathing in old mice compared to controls. Male 22-month old C57BL/6 J mice consumed a diet containing capsaicin (50 ppm) or lecithin control for one month. Breathing patterns were obtained prior to/following the dietary supplementation period using unrestrained barometric plethysmography. Frequency, tidal volume (VT), minute ventilation (VE), VE to expelled carbon dioxide ratio (VE/VCO2) and VT divided by inspiratory time (VT/Ti) were analyzed at baseline and during a 15-minute hypoxic exposure (10% O2). Capsaicin supplemented mice showed greater VE, VE/VCO2 and TV/Ti during hypoxic exposure compared to controls, with no change at baseline. Overall, these findings suggest an acute augmented response to hypoxia following capsaicin administration in older mice.

Arterial blood sampling in male CD-1 and C57BL/6J mice with 1% isoflurane is similar to awake mice.

Isoflurane (ISO) is a commonly used anesthetic that offers rapid recovery for laboratory animal research. Initial studies indicated no difference in arterial Pco2 ( PaCO2 ) or pH between conscious (NO ISO) and 1% ISO-exposed CD-1 mice. Our laboratory investigated whether arterial blood sampling with 1% ISO is a suitable alternative to NO ISO sampling for monitoring ventilation in a commonly studied mouse strain. We hypothesized similar blood chemistry, breathing patterns, and cardiovascular responses with NO ISO and 1% ISO. C57BL/6J mice underwent unrestrained barometric plethysmography to quantify the pattern of breathing. Mice exposed to hypoxic and hypercapnic gas under 1% ISO displayed blunted responses; with air, there were no breathing differences. Blood pressure and heart rate were not different between NO ISO and 1% ISO-exposed mice breathing air. Oxygen saturation was not different between groups receiving 2% ISO, 1% ISO, or air. Breathing frequency stabilized at ~11 min of 1% ISO following 2% ISO exposure, suggesting that 11 min is the optimal time for a sample in C57BL/6J mice. Blood samples at 1% ISO and NO ISO revealed no differences in blood pH and PaCO2 in C57BL/6J mice. Overall, this method reveals similar arterial blood sampling values in awake and 1% ISO CD-1 and C57BL/6J mice exposed to air. Although this protocol may be appropriate in other mouse strains when a conscious sample is not feasible, caution is warranted first to identify breathing frequency responses at 1% ISO to tailor the protocol. NEW & NOTEWORTHY Conscious arterial blood sampling is influenced by extraneous factors and is a challenging method due to the small size of mice. Through a series of experiments, we show that arterial blood sampling with 1% isoflurane (ISO) is an alternative to awake sampling in C57BL/6J and CD-1 male mice breathing air. Monitoring breathing frequency during 1% ISO is important to the protocol and should be closely followed to confirm adequate recovery after the catheter implantation.

Durability of collagen injection for stress urinary incontinence in women proven by transvaginal 3-dimensional ultrasound.

OBJECTIVE: The aim of this study is to evaluate the durability of collagen injection (CI) using serial 3-dimensional (3D) transvaginal ultrasound (US) in women with sufficient improvement of stress urinary incontinence (SUI) symptoms not requiring additional treatment. METHODS: After the institutional review board approval, a retrospective review of prospectively followed patients who underwent CI was conducted. Eligible patients received 3 or less consecutive CI for SUI with no reinjection afterward during the follow-up period and had a minimum follow-up of 1 year from the last CI. Serial 3D US was obtained for collagen volume and configuration at baseline (6-8 weeks postoperatively) and approximately every 12 months thereafter as clinically indicated. RESULTS: Of 191 eligible patients from 1/99 to 6/11, 67 (35%) met the inclusion criteria. The mean age was 67 years (42-90 years) with mean follow-up from the time of last CI at 43 months (12-149 months). A total of 283 three-dimensional US were performed, with the mean of 4 (2-11) per patient. The mean number of injections was 1.4 (1-3) with a mean injected volume at 5.8 mL (2-18 mL). The volume retention rate compared with the baseline volume was 84% (12%-100%), with a decrease in mean collagen volumes between the baseline and last follow-up visit (3.2 vs 2.7 mL; P = 0.008). Collagen volume decreased by a mean of 0.11 mL for each year past the final injection (P = 0.0015) by mixed-effect model analysis. CONCLUSIONS: Although believed to be nondurable, CI was found to be objectively stable over time by transvaginal 3D US in a subset of women with durably improved SUI symptoms.