Different Levels Of Hypoxia Research Articles

Activated Endothelium Changes The Activity Of Multipotent Mesenchymal Stromal Cells During Physiological Hypoxia Or Short Hypoxic Stress In Vitro

Multipotent mesenchymal stromal cells (MSCs) are used for supplemental therapy of ischemic and inflammatory diseases. After systemic administration, transmigration of MSCs to the target tissue is accompanied by interaction with activated endothelial cells (ECs) at the site of injury. In this study, we investigated the influence of TNF-α-activated ECs on the functions of MSCs under different levels of hypoxia. For this purpose, MSCs and TNF-α activated ECs were cocultured in a direct cell-to-cell setting for a short period of time. MSCs retained their stromal phenotype and multilineage differentiation potential after interaction with activated ECs. At the same time, changes in molecules involved in MSC-cell and MSC-extracellular matrix interaction were detected. The paracrine activity of MSCs and activated ECs after interaction was demonstrated by both upregulated transcription and increased levels of pleiotropic IL-6 and IL-8. Proteases/antiproteases profiles were also altered after interaction. These data suggest that short-term interaction of MSCs with activated ECs may play an important role in tissue repair and remodeling processes. In particular, it may promote the migratory phenotype of MSCs. In comparison to physiological hypoxia – 5% O2, acute hypoxic stress (0.1% O2, 24 h) attenuated the stimulatory effects of ECs on MSCs.

Prognostic genome and transcriptome signatures in colorectal cancers

Colorectal cancer is caused by a sequence of somatic genomic alterations affecting driver genes in core cancer pathways1. Here, to understand the functional and prognostic impact of cancer-causing somatic mutations, we analysed the whole genomes and transcriptomes of 1,063 primary colorectal cancers in a population-based cohort with long-term follow-up. From the 96 mutated driver genes, 9 were not previously implicated in colorectal cancer and 24 had not been linked to any cancer. Two distinct patterns of pathway co-mutations were observed, timing analyses identified nine early and three late driver gene mutations, and several signatures of colorectal-cancer-specific mutational processes were identified. Mutations in WNT, EGFR and TGFβ pathway genes, the mitochondrial CYB gene and 3 regulatory elements along with 21 copy-number variations and the COSMIC SBS44 signature correlated with survival. Gene expression classification yielded five prognostic subtypes with distinct molecular features, in part explained by underlying genomic alterations. Microsatellite-instable tumours divided into two classes with different levels of hypoxia and infiltration of immune and stromal cells. To our knowledge, this study constitutes the largest integrated genome and transcriptome analysis of colorectal cancer, and interlinks mutations, gene expression and patient outcomes. The identification of prognostic mutations and expression subtypes can guide future efforts to individualize colorectal cancer therapy.

Analysis of microRNA expression in rat kidneys after VEGF inhibitor treatment under different degrees of hypoxia.

Previously, we found that the incidence of kidney injury in patients with chronic hypoxia was related to the partial pressure of arterial oxygen. However, at oxygen concentrations that contribute to kidney injury, the changes in the relationship between microRNAs (miRNAs) and the hypoxia-inducible factor-1α (HIF-1α)-vascular endothelial growth factor (VEGF) axis and the key miRNAs involved in this process have not been elucidated. Therefore, we elucidated the relationship between VEGF and kidney injury at different oxygen concentrations and the mechanisms mediated by miRNAs. Sprague-Dawley rats were exposed to normobaric hypoxia and categorized into six groups based on the concentration of the oxygen inhaled and injection of the angiogenesis inhibitor bevacizumab, a humanized anti-VEGF monoclonal antibody. Renal tissue samples were processed to determine pathological and morphological changes and HIF-1α, VEGF, and miRNA expression. We performed a clustering analysis of high-risk pathways and key hub genes. The results were validated using two Gene Expression Omnibus datasets (GSE94717 and GSE30718). As inhaled oxygen concentration decreased, destructive changes in the kidney tissues became more severe. Although the kidney possesses a self-protective mechanism under an intermediate degree of hypoxia (10% O2), bevacizumab injections disrupted this mechanism, and VEGF expression was associated with the ability of the kidney to repair itself. rno-miR-124-3p was identified as a crucial miRNA; a key gene target, Mapk14, was identified during this process. VEGF plays an important role in kidney protection from injury under different hypoxia levels. Specific miRNAs and their target genes may serve as biomarkers that provide new insights into kidney injury treatment.NEW & NOTEWORTHY Renal tolerance to hypoxic environments is limited, and the degree of hypoxia does not show a linear relationship with angiogenesis. VEGF plays an important role in the kidney's self-protective mechanism under different levels of hypoxia. miR-124-3p may be particularly important in kidney repair, and it may modulate VEGF expression through the miR-124-3p/Mapk14 signaling pathway. These microRNAs may serve as biomarkers that provide new insights into kidney injury treatment.

Unveiling the hypoxia-induced mitophagy process through two-channel real-time imaging of NTR and viscosity under the same excitation

Mitophagy is an essential physiological process that eliminates damaged mitochondria via lysosomes. It is reported that hypoxia, inflammatory stimuli or other stress conditions could lead to mitochondrial damage and mitochondrial dysfunction, which induces the process of mitophagy. Herein, we report a novel fluorescent probe PC-NTR for imaging hypoxia-induced mitophagy by monitoring the change of nitroreductase and viscosity simultaneously. To our delight, PC-NTR could respond simultaneously to nitroreductase and viscosity at different fluorescence channels with no mutual interference under the same excitation wavelength. The fluorescence emission around 535 nm was enhanced dramatically after addition of nitroreductase while the fluorescence emission around 635 nm heightened as the viscosity increased. The probe would be able to selectively targeting of mitochondria in cells because of the positively charged pyridine salt structure of PC-NTR. The probe was successfully applied to assess the different levels of hypoxia and real-time imaging of mitochondrial autophagy in live cells. More importantly, using dual channel imaging, PC-NTR could be used to distinguish cancer cells from normal cells and was successfully applied to imaging experiments in HeLa-derived tumor-bearing nude mice. Therefore, PC-NTR would be an important molecular tool for hypoxia imaging and detecting solid tumors in vivo.

Effect of long-term hypoxia on the reproductive systems of female and male yellow catfish (Pelteobagrus fulvidraco).

This study investigated the effects of different levels of hypoxia on the reproductive system of yellow catfish. Yellow catfish (Pelteobagrus fulvidraco) were exposed to three dissolved oxygen concentration levels: normoxia (6.5 ± 0.2 mg/L), moderate hypoxia (MH, 3.8 ± 0.3 mg/L) and severe hypoxia (SH, 1.9 ± 0.2 mg/L) for 30 days. The gonadosomatic index of males, not females, significantly decreased in the SH group. In the SH group, for the females, the ratio of vitellogenic follicles significantly decreased, whereas the number of atretic follicles significantly increased. In male fish, a significantly reduced number of spermatozoa was observed in both the MH and SH groups. Elevated apoptosis levels in the testes and ovaries were observed only in the SH group. Serum 17β-estradiol and vitellogenin levels in females and testosterone levels in males significantly decreased in the SH group. The concentration of 11-ketotestosterone in males significantly decreased in both the MH and SH groups. In female fish, dysregulated expression of the hypothalamic-pituitary-gonadal (HPG) axis, steroidogenesis genes, and hepatic genes related to vitellogenesis were observed only in the SH group. However, in male fish, moderate hypoxia altered the expression of HPG genes, including gnrh1, lhcgr, and amh. Moreover, the MH group significantly altered the expression of steroidogenesis genes like star, 17β-hsd, and cyp17a1. The results of this study suggest that severe hypoxia can cause reproductive defects in female and male yellow catfish. Moreover, the reproductive system of male yellow catfish is more sensitive to moderate hypoxia than that of female catfish. Our findings contribute to our understanding of the response of the teleost reproductive system to long-term hypoxia.

Orexin facilitates the hypoxic ventilatory response via the activation of corticotropin-releasing hormone neurons that project to the nucleus of the solitary tract

The paraventricular nucleus of the hypothalamus (PVN) contributes significantly to the peripheral chemoreflex-mediated respiratory and sympathetic responses to hypoxia, in part via reciprocal interactions with nucleus of the solitary tract (nTS). Previously we showed that orexin neurons in the perifornical hypothalamus are activated by hypoxia and facilitate the hypoxic ventilatory response (HVR) in the active phase of the circadian cycle . The neural circuitry through which orexin facilitates the HVR is unknown. Orexin receptors (OxR) are expressed in the PVN, and we recently showed that OxR blockade reduced the activation of both corticotropin-releasing hormone neurons in the PVN as well as nTS neurons. In addition, hypoxia activated orexin neurons projecting to the PVN, but not those projecting to nTS. Here we further explored the hypothesis that orexin facilitates the HVR and arterial blood pressure (ABP) response to hypoxia via the activation of nTS-projecting CRH neurons, and that specific OxR1 knockdown in the PVN would reduce the HVR. To test these hypotheses, we nanoinjected fluorescent retrobeads bilaterally into the nTS to label nTS-projecting neurons. Rats were then exposed to hypoxia (2hrs at 11% O2 during the active phase) following injection of either suvorexant (OxR antagonist; 20 mg/kg i.p.; n=4), or vehicle (DMSO; n=4). Using immunohistochemistry (IHC) we then quantified the number of activated (i.e., Fos-immunoreactive; IR) nTS-projecting CRH neurons. In separate groups of rats we nanoinjected shRNA to knockdown Ox1R expression specifically in the PVN (n=8). PVN of control rats was nanoinjected with scrambled RNA (scRNA; n=3) or aCSF (n=2). Three weeks later, rats were exposed to graded hypoxia (from 21% to 10% inspired O2) over 3-5 minutes to assess the acute HVR and ABP response to different levels of hypoxia. OxR blockade reduced the number of Fos-IR, nTS-projecting CRH neurons by ~32% (drug: p<0.0001). Images confirmed successful targeting of shRNA to PVN, and OxR1 knockdown (by ~50%) was confirmed in subsets of samples by Western Blot and IHC. Ox1R knockdown specifically in the PVN had no effect on resting ventilation, but significantly reduced the HVR (main effect: p=0.02; 22% decrease in VE/VCO2 at 13% O2), due to a reduction in the respiratory frequency response to hypoxia (main effect: p=0.004; 38 breath/min decrease at 13% O2). Ox1R knockdown also decreased ABP (~5 mmHg at all O2 levels; main effect: p=0.0015), but not its response to hypoxia. These data suggest that orexin neurons facilitate the HVR via OxR1 Grants: 5R01HL098602 (EH, DK), 5R01HL136710 (KC) This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

An Optochemical Oxygen Scavenger Enabling Spatiotemporal Control of Hypoxia.

We present an optochemical O2 scavenging system that enables precise spatiotemporal control of the level of hypoxia in living cells simply by adjusting the light intensity in the illuminated region. The system employs rhodamine containing a selenium or tellurium atom as an optochemical oxygen scavenger that rapidly consumes O2 by photochemical reaction with glutathione as a coreductant upon visible light irradiation (560-590 nm) and has a rapid response time, within a few minutes. The glutathione-consuming quantum yields of the system were calculated as about 5 %. The spatiotemporal O2 consuming in cultured cells was visualized with a hypoxia-responsive fluorescence probe, MAR. Phosphorescence lifetime imaging was applied to confirmed that different light intensities could generate different levels of hypoxia. To illustrate the potential utility of this system for hypoxia research, we show that it can spatiotemporally control calcium ion (Ca2+ ) influx into HEK293T cells expressing the hypoxia-responsive Ca2+ channel TRPA1.

An Optochemical Oxygen Scavenger Enabling Spatiotemporal Control of Hypoxia

AbstractWe present an optochemical O2 scavenging system that enables precise spatiotemporal control of the level of hypoxia in living cells simply by adjusting the light intensity in the illuminated region. The system employs rhodamine containing a selenium or tellurium atom as an optochemical oxygen scavenger that rapidly consumes O2 by photochemical reaction with glutathione as a coreductant upon visible light irradiation (560–590 nm) and has a rapid response time, within a few minutes. The glutathione‐consuming quantum yields of the system were calculated as about 5 %. The spatiotemporal O2 consuming in cultured cells was visualized with a hypoxia‐responsive fluorescence probe, MAR. Phosphorescence lifetime imaging was applied to confirmed that different light intensities could generate different levels of hypoxia. To illustrate the potential utility of this system for hypoxia research, we show that it can spatiotemporally control calcium ion (Ca2+) influx into HEK293T cells expressing the hypoxia‐responsive Ca2+ channel TRPA1.

Two Color Imaging of Different Hypoxia Levels in Cancer Cells.

Hypoxia (low oxygen levels) occurs in a range of biological contexts, including plants, bacterial biofilms, and solid tumors; it elicits responses from these biological systems that impact their survival. For example, conditions of low oxygen make treating tumors more difficult and have a negative impact on patient prognosis. Therefore, chemical probes that enable the study of biological hypoxia are valuable tools to increase the understanding of disease-related conditions that involve low oxygen levels, ultimately leading to improved diagnosis and treatment. While small-molecule hypoxia-sensing probes exist, the majority of these image only very severe hypoxia (<1% O2) and therefore do not give a full picture of heterogeneous biological hypoxia. Commonly used antibody-based imaging tools for hypoxia are less convenient than small molecules, as secondary detection steps involving immunostaining are required. Here, we report the synthesis, electrochemical properties, photophysical analysis, and biological validation of a range of indolequinone-based bioreductive fluorescent probes. We show that these compounds image different levels of hypoxia in 2D and 3D cell cultures. The resorufin-based probe 2 was activated in conditions of 4% O2 and lower, while the Me-Tokyo Green-based probe 4 was only activated in severe hypoxia─0.5% O2 and less. Simultaneous application of these compounds in spheroids revealed that compound 2 images similar levels of hypoxia to pimonidazole, while compound 4 images more extreme hypoxia in a manner analogous to EF5. Compounds 2 and 4 are therefore useful tools to study hypoxia in a cellular setting and represent convenient alternatives to antibody-based imaging approaches.

Proton Tumor Control Relative Biological Effectiveness (RBE) Estimated for Various Fractionation Schedules and Hypoxia Levels

<h3>Purpose/Objective(s)</h3> We previously modeled clinical proton local control data of early-stage lung cancer using a simulation model based on an energy budget. The fitted modeling results suggest that proton therapy is nearly independent of hypoxia reoxygenation which is a key process to model photon therapy results. In this work, we investigated the effect of variable hypoxia levels on the local control efficacy of proton therapy compared to photon, in terms of RBE, for various fractionation schedules, searching for the optimal treatment option. <h3>Materials/Methods</h3> Model simulations were performed with a previously developed tumor simulation model with clinically derived parameter values for photon (CCR; 23; 5469-79; 2017) and proton (presented at RRS 2021). For five different levels of hypoxia, represented by growth fractions (GF), where lower GF represents more hypoxic condition, simulations were performed for 14 clinically applied fractionation schedules (see Table). For each proton schedule, the cell survival fraction was estimated and a partner simulation was performed with photon parameters, to find the dose level with the same fractionation scheme that would lead to an equivalent cell survival level. Finally, the proton RBE was calculated from the ratio of determined photon physical dose to physical dose of proton. <h3>Results</h3> The estimated proton RBE depends on both the hypoxia level as well as the fractionation schedule (see Table). The proton RBE increases with increasing hypoxic level (decreasing GF) for all fractionation schedules. For shorter fractionation schedules (3-5 fxs), proton therapy is always more effective than photon, with RBEs of 1.06-1.18. For longer fractionation schedules (>10), however, the RBE decreases as the GF increases (less hypoxia) and even drops below 1.0 (down to 0.81) for some schedules, due to the ability of photon therapy to take advantage of long term reoxygenation. For each hypoxia level, an optimal schedule exists with the highest RBE value: longer schedules (10-12 fx) are better for hypoxic tumors; 5 fx schedules are better for less hypoxic tumors. <h3>Conclusion</h3> The results predict that proton vs. photon local control will depend critically on the hypoxic level, with proton therapy more effective as the level of hypoxia increases. For longer treatment schedules, proton therapy loses this advantage with decreasing hypoxia, suggesting that longer proton schedules may be less effective for well-oxygenation tumors. These predicted effects should be further probed through both preclinical experiments and clinical data analyses.

The Rate of Cisplatin Dosing Affects the Resistance and Metastatic Potential of Triple Negative Breast Cancer Cells, Independent of Hypoxia

To best control tumor growth and/or metastasis in triple negative breast cancer (TNBC), it may be useful to understand the effect(s) of chemotherapy delivery (i.e., the rate and pattern of exposure to the drug) on cell sub-populations that have experienced different levels of hypoxia (and/or acidosis). In this spirit, MDA-MB-231 TNBC cells, and their hypoxia-reporter counterparts, were characterized for their sensitivity to cisplatin. When in the form of multicellular spheroids, that capture the diffusion-limited transport that generates hypoxic and acidic subregions within the avascular areas of solid tumors, the effects of the rate and pattern of exposure to cisplatin on cell viability and motility/migration potential were evaluated for each cell sub-population. We demonstrated that cell sensitivity to cisplatin was not dependent on acidosis, but cell resistance increased with exposure to hypoxia. In spheroids, the increase of the rates of cell exposure to cisplatin, at a constant cumulative dose, increased sensitivity to chemotherapy and lowered the cells’ metastatic potential, even for cells that had experienced hypoxia. This effect was also shown to be caused by nanocarriers engineered to quickly release cisplatin which deeply penetrated the spheroid interstitium, resulting in the fast and uniform exposure of the TNBC tumors to the agent. This rate and dosing-controlled model may effectively limit growth and/or metastasis, independent of hypoxia. This mode of chemotherapy delivery can be enabled by engineered nanocarriers.

CYP1A1, VEGFA and Adipokine Responses of Human Adipocytes Co-exposed to PCB126 and Hypoxia.

It is increasingly recognized that hypoxia may develop in adipose tissue as its mass expands. Adipose tissue is also the main reservoir of lipophilic pollutants, including polychlorinated biphenyls (PCBs). Both hypoxia and PCBs have been shown to alter adipose tissue functions. The signaling pathways induced by hypoxia and pollutants may crosstalk, as they share a common transcription factor: aryl hydrocarbon receptor nuclear translocator (ARNT). Whether hypoxia and PCBs crosstalk and affect adipokine secretion in human adipocytes remains to be explored. Using primary human adipocytes acutely co-exposed to different levels of hypoxia (24 h) and PCB126 (48 h), we observed that hypoxia significantly inhibits the PCB126 induction of cytochrome P450 (CYP1A1) transcription in a dose-response manner, and that Acriflavine (ACF)—an HIF1α inhibitor—partially restores the PCB126 induction of CYP1A1 under hypoxia. On the other hand, exposure to PCB126 did not affect the transcription of the vascular endothelial growth factor-A (VEGFA) under hypoxia. Exposure to hypoxia increased leptin and interleukin-6 (IL-6), and decreased adiponectin levels dose-dependently, while PCB126 increased IL-6 and IL-8 secretion in a dose-dependent manner. Co-exposure to PCB126 and hypoxia did not alter the adipokine secretion pattern observed under hypoxia and PCB126 exposure alone. In conclusion, our results indicate that (1) hypoxia inhibits PCB126-induced CYP1A1 expression at least partly through ARNT-dependent means, suggesting that hypoxia could affect PCB metabolism and toxicity in adipose tissue, and (2) hypoxia and PCB126 affect leptin, adiponectin, IL-6 and IL-8 secretion differently, with no apparent crosstalk between the two factors.

Sprint Interval Exercise Improves Cognitive Performance Unrelated to Postprandial Glucose Fluctuations at Different Levels of Normobaric Hypoxia.

Objective: The aim of our study was to examine cognition response to sprint interval exercise (SIE) against different levels of hypoxia. Research design and methods: 26 recreational active males performed SIE (20 × 6 s of all-out cycling bouts, 15 s of passive recovery) under normoxia (FIO2: 0.209), moderate hypoxia (FIO2: 0.154), and severe hypoxia (FIO2: 0.112) in a single-blinded crossover design. Cognitive function and blood glucose were assessed before and after 0, 10, 30, and 60 min of the SIE. Heart rate (HR), peripheral oxygen saturation (SpO2), and ratings of perceived exertion (RPE, the Borg 6–20-point scale) during each SIE trial were recorded before and immediately after every five cycling bouts, and after 0, 10, 30, and 60 min of the SIE. Results: All the three SIE trials had a significantly faster overall reaction time in the Stroop test at 10 min after exercise as compared to that of the baseline value (p = 0.003, ƞ2 = 0.606), and returned to normal after 60 min. The congruent RT at 10 min after SIE was significantly shorter than that of the baseline (p < 0.05, ƞ2 = 0.633), while the incongruent RT at both 10 min and 30 min were significantly shorter than that measured at baseline (p < 0.05, ƞ2 = 0.633). No significant differences in terms of accuracy were found across the three trials at any time points (p = 0.446, ƞ2 = 0.415). Blood glucose was significantly reduced at 10 min and was sustained for at least 60 min after SIE when compared to pre-exercise in all trials (p < 0.05). Conclusions: Acute SIE improved cognitive function regardless of oxygen conditions, and the sustained improvement following SIE could last for at least 10–30 min and was unaffected by the altered blood glucose level.

The Acute Effects of Normobaric Hypoxia on Strength, Muscular Endurance and Cognitive Function: Influence of Dose and Sex.

Simple SummaryHypoxic training is a novel method to increase resistance training adaptations. There is some evidence that resistance exercise performed in systemic hypoxia can lead to structural and functional adaptations of skeletal muscle. Studies have also demonstrated that normobaric hypoxia (i.e., normal pressure, low oxygen) increases intramuscular metabolic stress and type two (fast twitch) fiber recruitment, which leads to a greater morphological adaptations over time. However, to date, there is no research that has investigated the effects of different doses of acute normobaric hypoxia on strength and muscular endurance performance, nor has there been research investigating potential sex-based differences.The aim of this study was to examine the acute effects of different levels of hypoxia on maximal strength, muscular endurance, and cognitive function in males and females. In total, 13 males (mean ± SD: age, 23.6 ± 2.8 years; height, 176.6 ± 3.9 cm; body mass, 76.6 ± 2.1 kg) and 13 females (mean ± SD: age, 22.8 ± 1.4 years; height, 166.4 ± 1.9 cm; body mass, 61.6 ± 3.4 kg) volunteered for a randomized, double-blind, crossover study. Participants completed a one repetition strength and muscular endurance test (60% of one repetition maximum to failure) for squat and bench press following four conditions; (i) normoxia (900 m altitude; FiO2: 21%); (ii) low dose hypoxia (2000 m altitude; FiO2: 16%); (iii) moderate dose hypoxia (3000 m altitude; FiO2: 14%); and (iv) high dose hypoxia (4000 m altitude; FiO2: 12%). Heart rate, blood lactate, rating of perceived exertion, and cognitive function was also determined during each condition. The one repetition maximum squat (p = 0.33) and bench press (p = 0.68) did not differ between conditions or sexes. Furthermore, squat endurance did not differ between conditions (p = 0.34). There was a significant decrease in bench press endurance following moderate (p = 0.02; p = 0.04) and high (p = 0.01; p = 0.01) doses of hypoxia in both males and females compared to normoxia and low dose hypoxia, respectively. Cognitive function, ratings of perceived exertion, and lactate were also significantly different in high and moderate dose hypoxia conditions compared to normoxia (p < 0.05). Heart rate was not different between the conditions (p = 0.30). In conclusion, high and moderate doses of acute normobaric hypoxia decrease upper body muscular endurance and cognitive performance regardless of sex; however, lower body muscular endurance and maximal strength are not altered.

Effects of different levels of hypoxia and hypercarbia on ventilation and gas exchange in Boa constrictor amaralis and Crotalus durissus (Squamata: Serpentes)

Ventilation and gas exchange have been studied in relatively few species of snakes, especially regarding their response to environmental hypoxia or hypercarbia. We exposed Crotalus durissus (N = 6) and Boa constrictor (N = 6) to decreasing levels of oxygen (12, 9, 6, 3 % O2) and increasing levels of carbon dioxide (1.5, 3.0, 4.5, 6.0 % CO2) and analyzed the effect of the different gas mixtures on ventilation and gas exchange using open-flow respirometry. Neither hypoxia nor hypercarbia significantly altered the duration of expiration or inspiration, nor their proportions. Both hypoxia and hypercarbia increased minute ventilation, but the decrease in oxygen had a less pronounced effect on ventilation. Gas exchange under normoxic conditions was low and was not significantly affected by hypoxia, but hypercarbia decreased gas exchange significantly in both species. While B. constrictor maintained its respiratory exchange ratio (RER) under hypercarbia between 0.5 and 1.0, C. durissus showed a RER above 1.0 during hypercarbia, due to a significantly greater CO2 excretion. The overall responses of both species to hypercarbia and especially to hypoxia were very similar, which could be associated to similar lifestyles as ambush hunting sit-and-wait predators that are able to ingest large prey items. The observed differences in gas exchange could be related to respiratory systems with macroscopically different structures, possessing only a tracheal lung in C. durissus, but two functional lungs in B. constrictor.

Effect of oxygen levels in cellular activity

Energy for life is produced inside the internal human energy factory, the mitochondrion, where low oxygen levels are likely to be correlated with stressful behaviors. These include thriving for performance, competition and perfection. One way to build a sustainable society is to reduce the daily stress, allowing human energy to target the control of energy resources and environmental safeguarding. So, it is important to assess the impact of decreased oxygen (hypoxia) in the body, especially in the brain, as a small imbalance in brain activity can affect vital energy production essential for sustainable life activities.This work addressed fluorescence changes associated with the formation of FAD and of reactive oxygen species (ROS), evoked by different levels of hypoxia. Exposing the slices to 0% and 21% O2, caused an increase in FAD autofluorescence. Upon reoxygenation (95% O2) the 0% O2 trace did not recover, while the 21% O2 signal decreased reaching the baseline. In contrast, at 40% and 65% O2 the FAD fluorescence decreased reversibly. Regarding ROS fluorescence, no significant changes were observed during the application of 0% and 21% O2. However, at 40% and 65% O2, the signals were enhanced recovering partially when the initial oxygen level (95% O2) was reintroduced.

Relationship between Decreased Oxygenation during Acute Hypoxia and Cognitive Deterioration in Healthy Humans

BackgroundDespite improvements in military aircraft safety, many aviation incidents are attributed to hypoxia. The correlation between moderate hypoxia and cognitive function are well studied but results remain controversial. Comparatively, the effects of severe hypoxia are still not well understood. In the present study, we examined the executive cognitive function and physiological indices under both moderate and severe hypoxia conditions.PurposeTo examine the relationship between executive function and physiological indices during acute hypoxia exposure.MethodsTen young healthy participants (4M/6F, 29±4 years) completed two different levels of hypoxia: 1) 10 minute moderate hypoxia (MH; FiO2= 0.118) and 2) 3 minute severe hypoxia (SH; FiO2= 0.077) following 3 minute baseline (BL; FiO2=0.209). Reaction time and errors of commission (defined as a response to a non‐target stimulus) were evaluated continuously using a custom Go/No‐go test at baseline and throughout hypoxia exposure. In this Go/No‐go test, participants were required to push a key when upper case “A” or “@” appeared but not when lower case “a” appeared on a computer monitor as quick and as accurately as possible. The stimuli were presented one at a time at any one of nine differing locations across the screen. Cerebral oxygenation (rSO2) was continuously monitored with near‐infrared spectroscopy (NIRS) positioned above the left supraorbital ridge. Finger oxygen saturation (SpO2) was continuously assessed using a pulse oximetry. Data were analyzed in every 30 seconds and are presented as mean±SE. Two‐way ANOVA and Bonferroni test for multiple comparisons was used to evaluate significant differences.ResultsBoth SpO2 and rSO2 were reduced with SH (SpO2 %, BL: 99.3±0.1% vs. SH: 70.7±1.5%, P&lt;0.01; rSO2 %, BL: 70±4% vs. SH: 58±4%, P&lt;0.01). In SH, correlations of SpO2 and rSO2 were significant to the number of errors during the cognitive performance task (SpO2; R= −0.65, rSO2; R= −0.58, Both P&lt;0.01). SH significantly increased errors of commission of the last 30 seconds in 3 minute hypoxic exposure. (BL: 0.6±0.2 errors vs. SH: 1.7±0.5 errors, P&lt;0.01). SH significantly decreased reaction time over hypoxic exposure (BL: 0.43±0.01 sec vs. SH: 0.41±0.01 sec, P&lt;0.01). Comparatively, MH did not elicit an increase in the number of errors of commission nor a decrease in reaction time (Both, P&gt;0.05).ConclusionExecutive function deteriorated during severe hypoxia. Oxygenation of the prefrontal lobe which controls inhibition may be related to the increase in commission errors. Devices that monitor physiological indices such as SpO2 or rSO2 might be beneficial for a model to predict executive deterioration.Support or Funding InformationThis project was funded by the Office of Naval Research (ONR), grant number N00014‐18‐D‐7001‐TO1.

In vivo imaging of hypoxia generation stimulated by testosterone using a micelle-based near-infrared fluorescent probe

Hypoxia generation is caused by insufficient oxygen (O2) in aggressively proliferating cancer cells or tumors, which can lead to resistance to chemotherapy and radiotherapy. However, current hypoxic probes have been evaluated only in artificially induced hypoxic phenomena while ignoring latent stimuli. Testosterone, as a major stimulus of androgen, can cause hypoxia and hypoxia-inducible factor-1α generation during prostate cancer cell (LNCaP) proliferation or prostate cancer growth. Therefore, the improvement of hypoxia probes in the context of testosterone stimulation is critical. In response to this need, we have designed a micelle-based near-infrared fluorescent probe for imaging hypoxia stimulated by testosterone. In this study, we synthesized an amphiphilic copolymer, PEG−b−poly(MPC), in which fluorescent squarylium (SQ) dye and black hole quencher 2 (BHQ2) were enwrapped to form a PMPC@BHQ2/SQ probe. In this design, the fluorescence of the SQ dye is quenched by the BHQ2. In the context of hypoxia in this study, the BHQ2 was destroyed with the reduction of azo-reductase and the fluorescence of the SQ dye was recovered. This process is oxygen level-dependent, and the sensitivity of the probe is sufficient to distinguish different levels of hypoxia (<1% O2). We demonstrated the feasibility of the developed PMPC@BHQ2/SQ probe for investigating testosterone-induced hypoxia in both cells and in vivo. As far as we know, this paradigm is the first design of a rational strategy for the fluorescent imaging of hypoxia generation stimulated by testosterone. This work will contribute to hypoxia-related biomedical research.

Killing Many Birds With Two Stones: Hypoxia and Fibrosis Can Generate Ectopic Beats in a Human Ventricular Model.

During cardiac diseases many types of anatomical and functional remodeling of cardiac tissue can occur. In this work, we focus on two conditions: hypoxia and fibrosis, which are part of complex pathological modifications that take place in many cardiac diseases (hypertrophic cardiomyopathy, hypertensive heart disease, and recurrent myocardial infarction) and respiratory diseases (obstructive pulmonary disease, obstructive sleep apnea, and cystic fibrosis). Using computational models of cardiac electrophysiology, we evaluate if the interplay between hypoxia and fibrosis is sufficient to trigger cardiac arrhythmia. We study the mechanisms behind the generation of ectopic beats, an arrhythmic trigger also known as premature ventricular contractions (PVCs), in regions with high hypoxia and fibrosis. First, we modify an electrophysiological model of myocytes of the human left ventricle to include the effects of hypoxia. Second, diffuse fibrosis is modeled by randomly replacing cardiac myocytes by non-excitable and non-conducting cells. The Monte Carlo method is used to evaluate the probability of a region to generate ectopic beats with respect to different levels of hypoxia and fibrosis. In addition, we evaluate the minimum size of three-dimensional slabs needed to sustain reentries for different stimulation protocols. The observed mechanism behind the initiation of ectopic beats is unidirectional block, giving rise to sustained micro-reentries inside the region with diffuse fibrosis and hypoxia. In summary, our results suggest that hypoxia and fibrosis are sufficient for the creation of a focal region in the heart that generates PVCs.

Activation of voltage-dependent K+ channels strongly limits hypoxia-induced elevation of [Ca2+ ]i in rat carotid body glomus cells.

We studied the role of the large-conductance Ca2+ -activated K+ channel (BK) and voltage-dependent K+ channels (Kv) on [Ca2+ ]i responses to a wide range of hypoxia at different resting cell membrane potential (Em ). BK/Kv were mostly closed at rest in normoxia. BK/Kv became basally active when cells were depolarized by elevated [KCl]o (>12mm). Regardless of whether BK/Kv were closed or basally open, hypoxia-induced elevation of [Ca2+ ]i was enhanced 2- to 3-fold by inhibitors of BK/Kv. Hypoxia-induced elevation of [Ca2+ ]i was enhanced ∼2-fold by an inhibitor of Kv2, a major Kv in rat glomus cells. Hypoxia did not inhibit BK in inside-out patches. Our study supports a scheme in which activation of BK/Kv strongly limits the magnitude of hypoxia-induced [Ca2+ ]i rise, with Kv having a much greater effect than BK. Large-conductance KCa (BK) and other voltage-dependent K+ channels (Kv) are highly expressed in carotid body (CB) glomus cells, but their role in hypoxia-induced excitation is still not well defined and remains controversial. We addressed this issue by studying the effects of inhibitors of BK (IBTX) and BK/Kv (TEA/4-AP) on [Ca2+ ]i responses to a wide range of hypoxia at different levels of resting cell membrane potential (Em ). IBTX and TEA/4-AP did not affect the basal [Ca2+ ]i in isolated glomus cells bathed in 5mm KClo , but elicited transient increases in [Ca2+ ]i in cells that were moderately depolarized (11-20mV) by elevation of [KCl]o (12-20mm). Thus, BK and Kv were mostly closed at rest and activated by depolarization. Four different levels of hypoxia (mild, moderate, severe, anoxia) were used to produce a wide range of [Ca2+ ]i elevation (0-700nm). IBTX did not affect the rise in [Ca2+ ]i , but TEA/4-AP strongly (∼3-fold) enhanced [Ca2+ ]i rise by moderate and severe levels of hypoxia. Guangxitoxin, a Kv2 blocker, inhibited the whole-cell current by ∼50%, and enhanced 2-fold the [Ca2+ ]i rise elicited by moderate and severe levels of hypoxia. Anoxia did not directly affect BK, but activated BK via depolarization. Our findings do not support the view that hypoxia inhibits BK/Kv to initiate or maintain the hypoxic response. Rather, our results show that BK/Kv are activated as glomus cells depolarize in response to hypoxia, which then limits the rise in [Ca2+ ]i . Inhibition of Kv may provide a mechanism to enhance the chemosensory activity of the CB and ventilation.