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An Inexpensive Open-Source Chamber for Controlled Hypoxia/Hyperoxia Exposure

ABSTRACT: Understanding hypoxia/hyperoxia exposure requires either a high-altitude research facility or a chamber in which gas concentrations are precisely and reproducibly controlled. Hypoxia-induced conditions such as hypoxic-ischemic encephalopathy (HIE), obstructive or central apneas, and ischemic stroke present unique challenges for the development of models with acute or chronic hypoxia exposure. Many murine models exist to study these conditions; however, there are a variety of different hypoxia exposure protocols used across laboratories. Experimental equipment for hypoxia exposure typically includes flow regulators, nitrogen concentrators, and premix oxygen/nitrogen tanks. Commercial hypoxia/hyperoxia chambers with environmental monitoring are incredibly expensive and require proprietary software with subscription fees or highly expensive software licenses. Limitations exist in these systems as most are single animal systems and not designed for extended or intermittent hypoxia exposure. We have developed a simple hypoxia chamber with off-the-shelf components, and controlled by open-source software for continuous data acquisition of oxygen levels and other environmental factors (temperature, humidity, pressure, light, sound, etc.). Our chamber can accommodate up to two mouse cages and one rat cage at any oxygen level needed, when using a nitrogen concentrator or premixed oxygen/nitrogen tank with a flow regulator, but is also scalable. Our system uses a Python-based script to save data in a text file using modules from the sensor vendor. We utilized Python or R scripts for data analysis, and we have provided examples of data analysis scripts and acquired data for extended exposure periods (≤7 days). By using FLOS (Free-Libre and open-source) software and hardware, we have developed a low-cost and customizable system that can be used for a variety of exposure protocols. This hypoxia/hyperoxia exposure chamber allows for reproducible and transparent data acquisition and increased consistency with a high degree of customization for each experimenter’s needs.

SUBMITTER: Hillman T

PROVIDER: S-EPMC9315218 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Plant growth chambers produce controlled environments, which are crucial in making reproducible observations in experimental plant biology research. Commercial plant growth chambers can provide precise controls of environmental parameters, such as temperature, humidity, and light cycle, and the capability via complex programming to regulate these environmental parameters. But they are expensive. The high cost of maintaining a controlled growth environment is often a limiting factor when determining experiment size and feasibility. To overcome the limitation of commercial growth chambers, we designed and constructed an inexpensive plant growth chamber with consumer products for a material cost of $2,300. For a comparable growth space, a commercial plant growth chamber could cost $40,000 or more. Our plant growth chamber had outside dimensions of 1.5 m (W) x 1.8 m (D) x 2 m (H), providing a total growth area of 4.5 m2 with 40-cm high clearance. The dimensions of the growth area and height can be flexibly changed. Fluorescent lights with large reflectors provided a relatively spatially uniform photosynthetically active radiation intensity of 140-250 ?moles/m2/sec. A portable air conditioner provided an ample cooling capacity, and a cooling water mister acted as a powerful humidifier. Temperature, relative humidity, and light cycle inside the chamber were controlled via a z-wave home automation system, which allowed the environmental parameters to be monitored and programmed through the internet. In our setting, the temperature was tightly controlled: 22.2°C±0.8°C. The one-hour average relative humidity was maintained at 75%±7% with short spikes up to ±15%. Using the interaction between Arabidopsis and one of its bacterial pathogens as a test experimental system, we demonstrate that experimental results produced in our chamber were highly comparable to those obtained in a commercial growth chamber. In summary, our design of an inexpensive plant growth chamber will tremendously increase research opportunities in experimental plant biology.

Project description:Background: It was recently shown that intermittent hypoxic-hyperoxic exposure (IHHE) applied prior to a multimodal training program promoted additional improvements in cognitive and physical performance in geriatric patients compared to physical training only. However, there is a gap in the literature to which extent the addition of IHHE can enhance the effects of an aerobic training. Therefore, the aim of this study was to investigate the efficacy of IHHE applied prior to aerobic cycling exercise on cognitive and physical performance in geriatric patients. Methods: In a randomized, two-armed, controlled, and single-blinded trial, 25 geriatric patients (77-94 years) were assigned to two groups: intervention group (IG) and sham control group (CG). Both groups completed 6 weeks of aerobic training using a motorized cycle ergometer, three times a week for 20 min per day. The IG was additionally exposed to intermittent hypoxic and hyperoxic periods for 30 min prior to exercise. The CG followed the similar procedure breathing sham hypoxia and hyperoxia (i.e., normoxia). Within 1 week before and after the interventions, cognitive performance was assessed with the Dementia-Detection Test (DemTect) and the Clock Drawing Test (CDT), while physical performance was measured using the Timed "Up and Go" Test (TUG) and the Short-Physical-Performance-Battery (SPPB). Results: No interaction effect was found with respect to the DemTect (η p 2 = 0.02). An interaction effect with medium effect size (η p 2 = 0.08) was found for CDT performance with a higher change over time for IG (d = 0.57) compared to CG (d = 0.05). The ANCOVA with baseline-adjustment indicated between-group differences with a large and medium effect size at post-test for the TUG (η p 2 = 0.29) and SPPB (η p 2 = 0.06) performance, respectively, in favour of the IG. Within-group post-hoc analysis showed that the TUG performance was worsened in the CG (d = 0.65) and remained unchanged in the IG (d = 0.19). Furthermore, SPPB performance was increased (d = 0.58) in IG, but no relevant change over time was found for CG (d = 0.00). Conclusion: The current study suggests that an additional IHHE prior to aerobic cycling exercise seems to be more effective to increase global cognitive functions as well as physical performance and to preserve functional mobility in geriatric patients in comparison to aerobic exercise alone after a 6-week intervention period.

Project description:Preterm infants are highly susceptible to sustained lung inflammation, which may be triggered by exposure to multiple environmental cues such as supplemental oxygen (O2) and infections. We hypothesized that dysregulated macrophage (MФ) activation is a key feature leading to inflammation-mediated development of bronchopulmonary dysplasia (BPD) in preterm infants. Therefore, we aimed to determine age-dependent differences in immune responses of monocyte-derived MФ comparing cord blood samples derived from preterm (n=14) and term (n=19) infants as well as peripheral blood samples from healthy adults (n=17) after lipopolysaccharide (LPS) exposure. Compared to term and adult MФ, LPS-stimulated preterm MФ showed an enhanced and sustained pro-inflammatory immune response determined by transcriptome analysis, cytokine release inducing a RORC upregulation due to T cell polarization of neonatal T cells, and TLR4 surface expression. In addition, a double-hit model was developed to study pulmonary relevant exposure factors by priming MФ with hyperoxia (O2 = 65%) or hypoxia (O2 = 3%) followed by lipopolysaccharide (LPS, 100ng/ml). When primed by 65% O2, subsequent LPS stimulation in preterm MФ led to an exaggerated pro-inflammatory response (e.g. increased HLA-DR expression and cytokine release) compared to LPS stimulation alone. Both, exposure to 65% or 3% O2 together with subsequent LPS stimulation, resulted in an exaggerated pro-inflammatory response of preterm MФ determined by transcriptome analysis. Downregulation of two major transcriptional factors, early growth response gene (Egr)-2 and growth factor independence 1 (Gfi1), were identified to play a role in the exaggerated pro-inflammatory response of preterm MФ to LPS insult after priming with 65% or 3% O2. Preterm MФ responses to LPS and hyperoxia/hypoxia suggest their involvement in excessive inflammation due to age-dependent differences, potentially mediated by downregulation of Egr2 and Gfi1 in the developing lung.

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An Inexpensive Open-Source Chamber for Controlled Hypoxia/Hyperoxia Exposure

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