Computational tools for inversion and uncertainty estimation in respirometry.

In many physiological systems, real-time endogeneous and exogenous signals in living organisms provide critical information and interpretations of physiological functions; however, these signals or variables of interest are not directly accessible and must be estimated from noisy, measured signals. In this paper, we study an inverse problem of recovering gas exchange signals of animals placed in a flow-through respirometry chamber from measured gas concentrations. For large-scale experiments (e.g., long scans with high sampling rate) that have many uncertainties (e.g., noise in the observations or an unknown impulse response function), this is a computationally challenging inverse problem. We first describe various computational tools that can be used for respirometry reconstruction and uncertainty quantification when the impulse response function is known. Then, we address the more challenging problem where the impulse response function is not known or only partially known. We describe nonlinear optimization methods for reconstruction, where both the unknown model parameters and the unknown signal are reconstructed simultaneously. Numerical experiments show the benefits and potential impacts of these methods in respirometry.

Cigarette Smoke Triggers Loss of Corneal Endothelial Cells and Disruption of Descemet's Membrane Proteins in Mice.

Purpose: To investigate changes at a molecular level in the mouse corneal endothelium (CE) exposed to chronic cigarette smoke (CS). Methods: Pregnant mice (gestation days 18-20) were placed in a whole-body exposure smoking chamber, and a few days later pups were born. After 3.5 months of CS exposure, a ConfoScan4 scanning microscope was used to examine the corneal endothelial cells (CECs) of CS-exposed and control (Ct) mice. The CE was peeled under a microscope and maintained as four biological replicates (two male and two female) for CS-exposed and Ct mice; each replicate consisted of 16 CEs. The proteome of the CE was investigated through mass spectrometry. Results: The CE images of CS-exposed and Ct mice revealed a difference in the shape of CECs accompanied by a nearly 10% decrease in CEC density (P < 0.00003) following CS exposure. Proteome profiling identified a total of 524 proteins exhibiting statistically significant changes in CE from CS-exposed mice. Importantly, proteins associated with Descemet's membrane (DM), including COL4α1, COL4α2, COL4α3, COL4α4, COL4α5, COL4α6, COL8α1, COL8α2, and FN1, among others, exhibited diminished protein levels in the CE of CS-exposed mice. Conclusions: Our data confirm that exposure to CS results in reduced CEC density accompanied by diminished levels of multiple collagen and extracellular matrix proteins associated with DM.

Validity and reproducibility of VO2 max testing in a respiration chamber.

The aim of this study was to investigate whether VO2 max can be accurately measured in a respiration chamber. Thirty participants aged 23.4 ± 3.9 years with a wide range in VO2 max were included. Participants performed four incremental cycle ergometer tests (VO2 max) with a minimum of 5 days between tests. These tests consisted of one familiarization test with face mask, followed by two VO2 max tests in the respiration chamber and one test with face mask in randomized order. Oxygen consumption and CO2 production were measured continuously using Omnical (Maastricht University, the Netherlands) gas analysis system. The mean VO2 max was 3634 ± 766 ml, which resulted in mean VO2 max per lean body mass of 60.8 ± 8.0 ml/kg. Repeated respiration chamber tests showed a high concordance, and no significant differences were detected between tests (Lin's concordance correlation coefficient (Rc) = 0.99; ∆70 ± 302 ml/min; p = .38). There was high concordance between the mean VO2 max from both respiration chamber tests and the mean face mask tests, and no significant difference (Rc = 0.99; ∆41 ± 173 ml/min; p = .22) was observed. The Bland-Altman plots showed no proportional bias between different tests. In conclusion, the respiration chamber has been found to be a valid and reproducible method for measuring VO2 max. New research opportunities are possible in the respiration chamber, such as maximal exercise testing during 24-hour measurements.

Experimental Hypoxia as a Model for Cardiac Regeneration in Mice.

Experimental hypoxia has been used for decades to examine the adaptive response to low-oxygen environments. Various models have been studied, including flies, worms, fish, rodents, and humans. Our lab has recently used this technology to examine the effect of environmental hypoxia on mammalian heart regeneration. In this chapter, we describe studies of systemic hypoxia in mice. We found that systemic hypoxia can blunt oxidative DNA damage and induce cardiomyocyte proliferation. While our primary interests are focused on cardiovascular research, these hypoxia protocols are applicable to any other organ system.

The effects of energy metabolism variables on feed efficiency in respiration chamber studies with lactating dairy cows.

The objective of the present study was to investigate factors related to variation in feed efficiency (FE) among cows. Data included 841 cow/period observations from 31 energy metabolism studies assembled across 3 research stations. The cows were categorized into low-, medium-, and high-FE groups according to residual feed intake (RFI), residual energy-corrected milk (RECM), and feed conversion efficiency (FCE). Mixed model regression was conducted to identify differences among the efficiency groups in animal and energy metabolism traits. Partial regression coefficients of both RFI and RECM agreed with published energy requirements more closely than cofficients derived from production experiments. Within RFI groups, efficient (Low-RFI) cows ate less, had a higher digestibility, produced less methane (CH4) and heat, and had a higher efficiency of metabolizable energy (ME) utilization for milk production. High-RECM (most efficient) cows produced 6.0 kg/d more of energy-corrected milk (ECM) than their Low-RECM (least efficient) contemporaries at the same feed intake. They had a higher digestibility, produced less CH4 and heat, and had a higher efficiency of ME utilization for milk production. The contributions of improved digestibility, reduced CH4, and reduced urinary energy losses to increased ME intake at the same feed intake were 84, 12, and 4%, respectively. For both RFI and RECM analysis, increased metabolizability contributed to approximately 35% improved FE, with the remaining 65% attributed to the greater efficiency of utilization of ME. The analysis within RECM groups suggested that the difference in ME utilization was mainly due to the higher maintenance requirement of Low-RECM cows compared with Medium- and High-RECM cows, whereas the difference between Medium- and High-RECM cows resulted mainly from the higher efficiency of ME utilization for milk production in High-RECM cows. The main difference within FCE (ECM/DMI) categories was a greater (8.2 kg/d) ECM yield at the expense of mobilization in High-FCE cows compared with Low-FCE cows. Methane intensity (CH4/ECM) was lower for efficient cows than for inefficient cows. The results indicated that RFI and RECM are different traits. We concluded that there is considerable variation in FE among cows that is not related to dilution of maintenance requirement or nutrient partitioning. Improving FE is a sustainable approach to reduce CH4 production per unit of product, and at the same time improve the economics of milk production.

Effects of Chronic Nicotine Inhalation on Systemic and Pulmonary Blood Pressure and Right Ventricular Remodeling in Mice.

Cigarette smoking is the single most important risk factor for the development of cardiovascular and pulmonary diseases; however, the role of nicotine in the pathogenesis of these diseases is incompletely understood. The purpose of this study was to examine the effects of chronic nicotine inhalation on the development of cardiovascular and pulmonary disease with a focus on blood pressure and cardiac remodeling. Male C57BL6/J mice were exposed to air (control) or nicotine vapor (daily, 12 hour on/12 hour off) for 8 weeks. Systemic blood pressure was recorded weekly by radio-telemetry, and cardiac remodeling was monitored by echocardiography. At the end of the 8 weeks, mice were subjected to right heart catheterization to measure right ventricular systolic pressure. Nicotine-exposed mice exhibited elevated systemic blood pressure from weeks 1 to 3, which then returned to baseline from weeks 4 to 8, indicating development of tolerance to nicotine. At 8 weeks, significantly increased right ventricular systolic pressure was detected in nicotine-exposed mice compared with the air controls. Echocardiography showed that 8-week nicotine inhalation resulted in right ventricular (RV) hypertrophy with increased RV free wall thickness and a trend of increase in RV internal diameter. In contrast, there were no significant structural or functional changes in the left ventricle following nicotine exposure. Mechanistically, we observed increased expression of angiotensin-converting enzyme and enhanced activation of mitogen-activated protein kinase pathways in the RV but not in the left ventricle. We conclude that chronic nicotine inhalation alters both systemic and pulmonary blood pressure with the latter accompanied by RV remodeling, possibly leading to progressive and persistent pulmonary hypertension.

Nicotine e-cigarette vapor inhalation effects on nicotine & cotinine plasma levels and somatic withdrawal signs in adult male Wistar rats.

RATIONALE: Non-contingent chronic nicotine exposure procedures have evolved rapidly in recent years, culminating in electronic nicotine delivery systems (ENDS or e-cigarettes) to deliver vaporized drugs to rodents in standard housing chambers. OBJECTIVES: The aim of the current work was to use ENDS to test concentration-dependent effects of nicotine e-cigarette vapor inhalation on blood-nicotine concentrations, blood-cotinine concentrations, and somatic withdrawal signs over time in rats. METHODS: Male Wistar rats were exposed to vapor containing various concentrations of nicotine (20, 40, 80 mg/mL) for 11 days through ENDS, and blood concentrations of nicotine and cotinine, the major proximate metabolite of nicotine, as well as spontaneous and precipitated somatic withdrawal signs, were measured over time (across days of exposure and over hours after termination of vapor exposure). RESULTS: Exposing male Wistar rats to non-contingent nicotine vapor inhalation through ENDS produces somatic withdrawal symptoms and measurable blood-nicotine and blood-cotinine levels that change according to (1) concentration of nicotine in vape solution, (2) number of days of nicotine vapor exposure, (3) time since termination of nicotine vapor exposure, and (4) relative to the withdrawal signs, whether withdrawal was spontaneous or precipitated (by mecamylamine). CONCLUSIONS: The data presented here provide parameters that can be used as a reasonable starting point for future work that employs ENDS to deliver non-contingent nicotine vapor in rats, although many parameters can and should be altered to match the specific goals of future work.

Characterization of serum metabolites as biomarkers of carbon black nanoparticles-induced subchronic toxicity in rats by hybrid triple quadrupole time-of-flight mass spectrometry with non-targeted metabolomics strategy.

Carbon black nanoparticles (CBNPs) are one of atmospheric particles components and have been closely related with a series of lung diseases. It can reach the depths of the respiratory tract or even alveolar more easily than those micro-particles. Although some of its toxicities have been confirmed in animals or human bodies, the subchronic toxicity mechanism of CBNPs has been uncertain so far. Therefore, it is very necessary to establish a novel method and clarify the mechanism of subchronic toxicity caused by concentration adjustments of small molecule metabolites in vivo. In animal experiments, CB exposure, recovery and control group were set up. The concentration of CBNPs in chamber was 30.06 ±â€¯4.42 mg/m3. We developed a UHPLC-Q-TOF-MS/MS-based non-targeted metabolomic analysis strategy to analyze serum samples of rats. Then, differential metabolites in serum were found by multivariate data analysis and 39 potential biomarkers were identified. It was showed that main metabolic pathways associated with CBNPs exposure were hormones metabolism, amino acid metabolism, nucleotide metabolism and lipid metabolism. It is worth noting that long-term exposure to CBNPs had the greatest impact on steroid hormones biosynthesis so that the risk of infertility could increase. The results provided a new mechanistic insight into the metabolic alterations owing to CBNPs induced subchronic toxicity.

Measuring dynamic Eustachian tube function using tympanometry in a pressure chamber: the effect of nasal betahistine application.

OBJECTIVE: To assess the effect of topical betahistine on Eustachian tube function in subjectively abnormal subjects in a hyperbaric chamber. METHOD: Active and passive Eustachian tube function was examined using tympanometry in a pressure chamber. RESULTS: Active Eustachian tube function was tested against the negative middle ear pressure induced by increasing the chamber pressure to +3 kPa. One voluntary swallow decreased middle-ear pressure by a mean of 1.36 kPa. Passive Eustachian tube function was tested by measuring spontaneous Eustachian tube openings as the chamber pressure dropped from +10 kPa to ambient. Four distinct patterns of Eustachian tube behaviour were seen, three of which indicated Eustachian tube dysfunction. Betahistine had no positive effect on Eustachian tube opening, although previous animal studies had suggested a beneficial effect. CONCLUSION: Topical betahistine had no effect on Eustachian tube function. Combining a hyperbaric chamber with tympanometry proved ideal for evaluating Eustachian tube function.

Evaluation of the Parameter "Mean Impedance" for Representing Eustachian tube Functions During Pressure Increase and Decrease in Pressure Chamber Measurements.

HYPOTHESIS: The hypothesis of the study is that the mean impedance (MI) during compression and decompression provides additional information of the Eustachian tube (ET) function. BACKGROUND: The continuous impedance measurement in a pressure chamber can provide valuable information about the opening function of the ET. METHODS: Around 55 ear-healthy volunteers were examined in a pressure chamber. These were subjected to a decompression phase and a compression phase. The pressure change was constantly 20 kPa/min. Using evaluation software, the MI could be determined for both ears in each case for the phases of compression and decompression. RESULTS: In 49 participants, we could interpret the data successfully. On average, an output value (without pressure changes) of the impedance of 0.58 ±0.11 Pa on the right side and 0.43 ±â€Š0.1 Pa on the left side were measured. During decompression, 0.098 ±â€Š0.05 Pa (right) and 0.087 ±â€Š0.043 Pa (left) could be determined. For compression, values of 0.086 ±â€Š0.044 Pa on the right and 0.079 ±â€Š0.045 Pa on the left were detected. The retest reliability was higher with an intraclass correlation coefficient for the decompression MI of 0.833 than the 0.772 compression MI. CONCLUSIONS: It is possible to measure MI in healthy subjects during compression and decompression. This value represents a good average in terms of the pressure tolerance of the middle ear. In future, studies will be required to determine whether MI will be a useful parameter in differentiating normal and abnormal ET function.

Personalized medicine-concepts, technologies, and applications in inflammatory skin diseases.

The current state, tools, and applications of personalized medicine with special emphasis on inflammatory skin diseases like psoriasis and atopic dermatitis are discussed. Inflammatory pathways are outlined as well as potential targets for monoclonal antibodies and small-molecule inhibitors.

Development of a large-scale computer-controlled ozone inhalation exposure system for rodents.

Objective: Complete systems for laboratory-based inhalation toxicology studies are typically not commercially available; therefore, inhalation toxicologists utilize custom-made exposure systems. Here we report on the design, construction, testing, operation and maintenance of a newly developed in vivo rodent ozone inhalation exposure system. Materials and methods: Key design requirements for the system included large-capacity exposure chambers to facilitate studies with large sample sizes, automatic and precise control of chamber ozone concentrations, as well as automated data collection on airflow and environmental conditions. The exposure system contains two Hazelton H-1000 stainless steel and glass exposure chambers, each providing capacity for up to 180 mice or 96 rats. We developed an empirically tuned proportional-integral-derivative control loop that provides stable ozone concentrations throughout the exposure period (typically 3h), after a short ramp time (∼8 min), and across a tested concentration range of 0.2-2 ppm. Specific details on the combination of analog and digital input/output system for environmental data acquisition, control and safety systems are provided, and we outline the steps involved in maintenance and calibration of the system. Results: We show that the exposure system produces consistent ozone exposures both within and across experiments, as evidenced by low coefficients of variation in chamber ozone concentration and consistent biological responses (airway inflammation) in mice, respectively. Conclusion: Thus, we have created a large and robust ozone exposure system, facilitating future studies on the health effects of ozone in rodents.

Clinical standardization of two controlled allergen challenge facilities: The Environmental Exposure Unit and the Biogenics Research Chamber.

BACKGROUND: Controlled allergen challenge facilities (CACF), in disparate geographic regions with dissimilar engineering and base populations, have historically functioned as single, independent sites in clinical allergy trials. We aimed to demonstrate "between-unit reproducibility" to allow controlled challenge trials of participants using 2 CACFs. OBJECTIVE: To compare and standardize 2 CACFs located in Kingston, Ontario, Canada, and San Antonio, Texas, by examining participant-reported symptom severity during qualifying and treatment visits and evaluating response to treatment, while using the same allergen. METHODS: At 2 different CACFs, participants were enrolled in a double-blind, placebo-controlled, crossover intervention trial with cetirizine 10 mg. Different distribution devices delivered common short ragweed pollen via laminar air flow and maintained an airborne concentration of 3500 ± 700 grains/m3 in both facilities. A 1-hour "sham" run with no pollen release preceded a priming exposure of 3 hours and was followed 3 days later by a qualifying/treatment 5-hour exposure. At least 14 days later, another priming exposure was followed by the crossover exposure and treatment. RESULTS: Forty-eight and 43 subjects completed the study at Kingston and San Antonio, respectively. Demographics were similar. Fewer than 10% exhibited symptoms with sham exposure. No significant differences were found between the 2 facilities in maximal total rhinoconjunctivitis symptom score, total nasal symptom score, and total ocular symptom score, nor in areas under the curve. In both facilities, no significant effects of cetirizine 10 mg over placebo were detected. CONCLUSION: The results were equivalent, demonstrating that the 2 CACFs can be used together in dual-center clinical trials and show the possibility of multicenter trials involving multiple CACFs.

Lowering barometric pressure induces neuronal activation in the superior vestibular nucleus in mice.

Weather changes accompanied by decreases in barometric pressure are suggested to trigger meteoropathy, i.e., weather-related pain. We previously reported that neuropathic pain-related behavior in rats is aggravated by lowering barometric pressure, and that this effect is abolished by inner ear lesions. These results suggest that mechanisms that increase vestibular neuronal activity may parallel those that contribute to meteoropathy generation. However, it remains unknown whether changes in barometric pressure activate vestibular neuronal activity. To address this issue, we used expression of c-Fos protein as a marker for neural activation. Male and female mice were placed in a climatic chamber, and the barometric pressure was lowered by 40 hPa, from 1013 hPa, for 50 min (LP stimulation). The total number of c-Fos-positive cells in the vestibular nuclei was counted bilaterally after LP stimulation. We also video-recorded mouse behaviors and calculated the total activity score during the LP stimulation. LP stimulation resulted in significant c-Fos expression in the superior vestibular nucleus (SuVe) of male and female mice. There was no effect of LP stimulation on the total activity score. These data show that distinct neurons in the SuVe respond to LP stimulation. Similar mechanisms may contribute to the generation of meteoropathy in humans.

Chronic intermittent nicotine delivery via lung alveolar region-targeted aerosol technology produces circadian pharmacokinetics in rats resembling human smokers.

Cigarette smoke is an aerosol containing microparticles that carry nicotine into the lung alveolar region where nicotine is rapidly absorbed into circulation. Nicotine exposure in smokers is a chronic intermittent process, with episodic intake during wakefulness and abstinence during sleep resulting in circadian fluctuation of blood nicotine levels. We developed an integrated platform where freely moving rodents can be exposed to episodic nicotine aerosol on an investigator-designed schedule. Plasma nicotine and its metabolite cotinine levels were determined with a LC-MS/MS method. We characterized the aerosol in the breathing zone of the rodent exposure chamber. The droplet-size distribution was within the respirable diameter range. The system can generate a wide range of nicotine concentrations in air that meet a variety of experimental needs. Rats were exposed to nicotine aerosol once every half hour in the dark phase of 12:12-h light-dark cycles for 10 days. We optimized the parameters of aerosol generation and exposure: plasma nicotine and cotinine concentrations reached 30-35 and 190-240 ng/ml, respectively. The nicotine levels and circadian patterns resembled the pharmacokinetic pattern of human smokers. In summary, we developed an aerosol system that can produce clinically relevant chronic intermittent nicotine exposure in unanesthetized, unrestrained rodents with route of administration and circadian blood pharmacokinetics resembling human smokers. This methodology is a novel tool for understanding the health effects of chronic intermittent nicotine exposure such as with tobacco cigarettes and electronic cigarettes for studies of behavior, pharmacology and toxicology, nicotine addiction, tobacco-related diseases, and teratogenicity, and for the discovery of therapeutics. NEW & NOTEWORTHY We developed a lung alveolar region-targeted aerosol method and a system that provides chronic intermittent nicotine exposure in freely moving rodents. The method produces in rodents clinically relevant nicotine exposure with the route and circadian pharmacokinetics resembling human smokers. This method is a novel tool for understanding the health impacts of chronic nicotine exposures such as with tobacco cigarettes and electronic cigarettes, for studying nicotine pharmacology, toxicology, addiction, and tobacco-related diseases, and for the discovery of therapeutics.

Comparative study of the effect of bilastine and cetirizine on cognitive functions at ground level and at an altitude of 4,000 m simulated in hypobaric chamber: a randomized, double-blind, placebo-controlled, cross-over study.

INTRODUCTION: Antihistamines are easily accessible and cover the vast majority of the medical therapy of allergic rhinitis. However, their systemic administration may more frequently associate with their well-known side-effect, sedation, which is a serious problem in persons in safety-critical jobs such as aviation. We have a poor understanding whether a non-sedative antihistamine has an impact on vigilance when hypobaric hypoxia occurs during flight. METHODS: In this randomized, placebo-controlled, double-blind, cross-over study the effect of 20 mg bilastine was compared to 10 mg cetirizine and to placebo (20 mg pyridoxine) in 33 individuals at ground level and at 4,000 m altitude simulated in hypobaric chamber. Levels of vigilance, ultrashort memory, combined distributive attention, monotony tolerance and peripheral blood oxygen saturation (SpO2) were assessed. RESULTS: Bilastine did not impair the tested abilities in comparison with the control groups either at ground level or hypobaric hypoxia. Administration of cetirizine increased the number of errors at ground level. At the simulated altitude, already impaired results were additionally demonstrated with regards to the distributive attention test. CONCLUSIONS: From the two examined antihistamines, bilastine should be the preferred medication for by individuals who require constant attention and are exposed to hypobaric hypoxia.

[Environmental exposure chambers (EEC): A novel tool for pathophysiological and pharmaceutical research]. / Les chambres d'exposition environnementale (CEE) : un nouvel outil dans la recherche académique et industrielle.

Airborne allergic diseases (allergic asthma, rhinitis and conjunctivitis) have reached epidemic proportions and are a great burden for both society and individuals. Therefore we need to better understand the physiopathological mechanisms and to increase clinical research in these diseases. However, traditional outpatient studies are difficult and have number of limitations, in particular the variability of allergen exposure. Yet allergen provocation tests, especially bronchial challenges in asthma, are excellent tools to measure the efficiency of anti-allergic therapies. Environmental exposure chambers (EEC) allow the performance of controlled allergen provocation tests on a large scale with remarkable sensitivity, specificity and reproducibility. Moreover, they allow a precise collection of allergic symptoms, making them interesting tools for patho-physiological and clinical studies. During the last thirty years, they have assisted the study of anti-allergic therapies and provided data on their pharmacodynamic characteristics, particularly in allergic rhinitis. However, there are still no EEC tests centered on asthma. The EEC of Strasbourg (ALYATEC®) was developed to fulfill two objectives: to allow standardized allergenic and non-allergenic exposures with better control of the parameters than in other EEC and to offer a place to study asthma and anti-asthmatic therapies safely.

Onset of Action of the Fixed Combination Intranasal Azelastine-Fluticasone Propionate in an Allergen Exposure Chamber.

BACKGROUND: A fixed-dose combination of intranasal azelastine hydrochloride and fluticasone propionate (MP-AzeFlu) is the most effective treatment of allergic rhinitis, but its onset of action requires further investigation. OBJECTIVE: To compare the onset of action of MP-AzeFlu with the free combination of oral loratadine (LORA) and intranasal fluticasone propionate (INFP). METHODS: In this single-center, randomized, placebo-controlled, double-blind, double-dummy, 3-period crossover trial, allergic rhinitis symptoms were induced in asymptomatic patients by ragweed pollen challenge in an allergen environmental exposure chamber. Patients received single-dose MP-AzeFlu, LORA/INFP, or placebo and were monitored for 4 hours. The primary outcome was onset of action measured by total nasal symptom score (TNSS). Secondary measures were total ocular symptom score (TOSS), total score of the 7 nasal and ocular symptoms (T7SS), and the global visual analog scale (VAS). RESULTS: The full analysis set included 82 patients, of which 78 completed all treatments. TNSS was significantly reduced versus placebo from 5 minutes for MP-AzeFlu and 150 minutes for LORA/INFP onward (both P < .05) till the end of assessment (0-4 hours). MP-AzeFlu reduced TNSS to a greater extent at each time point from 5 to 90 minutes (P < .05) and over the entire assessment interval (P ≤ .005) versus LORA/INFP or placebo. No statistically significant difference between LORA/INFP and placebo was observed over the assessment interval (P = .182). The onset of action of MP-AzeFlu assessed by TOSS, T7SS, and VAS was 10 minutes, 2 hours earlier than with LORA/INFP. CONCLUSION: MP-AzeFlu had a more rapid onset of action (5 minutes) and was more effective than LORA/INFP.