kPa pO2 Research Articles

Effects of different oxygen regimes on ecological performance and bioenergetics of a coastal marine bioturbator, the soft shell clam Mya arenaria

Benthic species are exposed to oxygen fluctuations that can affect their performance and survival. Physiological effects and ecological consequences of fluctuating oxygen are not well understood in marine bioturbators such as the soft-shell clam Mya arenaria. We explored the effects of different oxygen regimes (21 days of exposure to constant hypoxia (~4.1 kPa PO2), cyclic hypoxia (~2.1–~10.4 kPa PO2) or normoxia (~21 kPa PO2)) on energy metabolism, oxidative stress and ecological behaviors (bioirrigation and bioturbation) of M. arenaria. Constant hypoxia and post-hypoxic recovery in cyclic hypoxia led to oxidative injury of proteins and lipids, respectively. Clams acclimated to constant hypoxia maintained aerobic capacity similar to the normoxic clams. In contrast, clams acclimated to cyclic hypoxia suppressed aerobic metabolism and activated anaerobiosis during hypoxia, and strongly upregulated aerobic metabolism during recovery. Constant hypoxia led to decreased lipid content, whereas in cyclic hypoxia proteins and glycogen accumulated during recovery and were broken down during the hypoxic phase. Digging of clams was impaired by constant and cyclic hypoxia, and bioirrigation was also suppressed under constant hypoxia. Overall, cyclic hypoxia appears less stressful for M. arenaria due to the metabolic flexibility that ensures recovery during reoxygenation and mitigates the negative effects of hypoxia, whereas constant hypoxia leads to depletion of energy reserves and impairs ecological functions of M. arenaria potentially leading to negative ecological consequences in benthic ecosystems.

Response to Different Oxygen Partial Pressures and Evolution Analysis of Apoptosis-Related Genes in Plateau Zokor (Myospalax baileyi)

The plateau zokor (Myospalax baileyi) is a native species of the Qinghai–Tibet Plateau that spends its entire life underground in sealed burrows with hypoxic conditions. The present study aimed to assess the sequence characteristics of apoptosis-related genes and the response to different oxygen partial pressures (pO2) in plateau zokor and Sprague-Dawley rats. The sequences of the p53-induced protein with a death domain (Pidd), p53-upregulated modulator of apoptosis (Puma), insulin-like growth factor binding protein 3 (Igfbp3), and apoptosis protease-activating factor 1 (Apaf1) were evaluated concerning homology and convergent evolution sites, and their mRNA levels were evaluated in different tissues under 14.13 (3,300 m) and 16.12 kPa (2,260 m) pO2 conditions. Our results showed that, (1) the sequences of the apoptosis-related genes in plateau zokor were highly similar to those of Nannospalax galili, followed by Rattus norvegicus; (2). Pidd, Puma, Igfbp3, and Apaf1 of plateau zokor were found to have five, one, two, and five convergent sites in functional domains with N. galili, respectively. Lastly (3), under low pO2, the expression of Pidd and Puma was downregulated in the lung of plateau zokors. In turn, Igfbp3 and Apaf1 were upregulated in the liver and lung, and Puma was upregulated in the skeletal muscle of plateau zokor under low pO2. In Sprague-Dawley rats, low pO2 downregulated Puma and Apaf1 expression in the liver and downregulated Igfbp3 and Puma in the lung and skeletal muscle separately. In contrast, low pO2 upregulated Pidd expression in the liver and skeletal muscle of Sprague-Dawley rats. Overall, the expression patterns of Apaf1, Igfbp3, and Puma showed the opposite pattern in the liver, lung, and skeletal muscle, respectively, of plateau zokor as compared with Sprague-Dawley rats. In conclusion, for the long-time adaptation to hypoxic environments, Pidd, Puma, Igfbp3, and Apaf1 of plateau zokor underwent convergent evolution, which we believe may have led to upregulation of their levels under low oxygen partial pressures to induce apoptosis, so as to suppress tumorigenesis under hypoxic environments in plateau zokor.

Low irradiance disrupts the internal O2 dynamics of seagrass (Thalassia testudinum) leading to shoot meristem H2S intrusion

Hypoxia and hydrogen sulfide (H2S) intrusion at night contribute to large-scale seagrass mortality events world-wide. Declining water quality has lowered irradiance and enhanced hypoxia in seagrass ecosystems, but linkages between low irradiance and seagrass internal pO2in situ are not well understood. We examined low irradiance effects on leaf and meristem pO2 dynamics of a dominant tropical seagrass, Thalassia testudinum, using microsensors over multiple diurnal cycles. Further, we determined how O2 dynamics affect H2S intrusion into shoot meristems under low irradiance. Sequential days of low irradiance disrupted internal leaf O2 status in three ways: i) causing a longer lag in morning O2 pressurization, ii) depressing maximum pO2 during the day, and iii) shortening the time where maximum pO2 was sustained into the afternoon. There was a close relationship between leaf and water column pO2 during the day (R2 = 0.93 ± 0.09); thus, internal O2 dynamics appear more dependent on water column pO2 under low irradiance. The nighttime minimum of leaf and meristem pO2 (1.2 and 0.4 kPa, respectively) were very low and the length of time the meristem sustained hypoxia (< 1.5 kPa pO2) at night was high (9:40 h:min). H2S intrusion into the meristem at night following 24–48 h of shading was persistent, likely the combined effect of limited internal O2 flow without saturating irradiances for photosynthesis, and a breakdown of the sediment oxic microshield. Under low irradiance, more frequent intrusions of H2S to the meristems increases the chances of large-scale seagrass mortality events at night.

Hypersalinity affects leaf and meristem O2 dynamics exposing meristems to H2S in the dominant tropical seagrass Thalassia testudinum

Large-scale (>80 km2) seagrass mortality events have been reported worldwide. While the mechanisms triggering these sudden die-off events are not well understood, plant hypoxia and H2S intrusion, at times linked to hypersaline conditions, have been suggested to play a role. In the present study, we used microsensors (~100 μm) to measure pO2 light-dark dynamics (4 and 1 h, light:dark) in the leaf, and simultaneously pO2 and H2S in the meristem, of the tropical seagrass, Thalassia testudinum. The role of leaf pO2 in mitigating H2S intrusion into the meristem was examined. We also assessed the interactive effect of water column hypoxia and hypersalinity (35, 45, 55, 65) on meristem pO2 light-dark dynamics and H2S intrusion. Internal gas dynamics were measured from seagrass shoots in intact cores collected at a site with recurrent die-off events in Florida Bay, USA. When the overlying water column pO2 was maintained at air equilibrium (20 kPa), the leaf pO2 in shoots at ≤55 salinity was supersaturated (30 to 35 kPa) with respect to the water column. However, leaves of shoots at 65 salinity in the light maintained leaf pO2 on par with the water column (~20–25 kPa), even after 4 h. This was the case even though the initial (10 min) leaf Δ pO2 was similar across salinity treatments, suggesting 65 salinity limited O2 pressurization, rather than, or in addition to, O2 production. The low leaf pO2 at 65 salinity in the light limited meristem pO2 (~10 kPa pO2) at the start of the dark cycle. Consequently, the meristem pO2 threshold of ~1.5 kPa pO2 was reached within 1–2 h of the dark cycle, and H2S intrusion occurred. Further, at 65 salinity, H2S intrusion occurred at higher water column pO2 (10–15 kPa), compared to salinity ≤55 (~5 kPa). Meristem minimum pO2 were also lower in the dark at higher salinity, indicating a greater belowground O2 demand. Based on these experimental microsensor results, we suggest two primary goals to minimize hypoxia and H2S-induced seagrass mortality: (1) minimize hypersaline conditions to increase the efficiency of seagrass leaf and meristem O2 pressurization, and (2) reduce nutrient loads and hypersaline conditions to keep water column pO2 at a maximum and lower sediment-meristem O2 demand in the dark.

Hypoxic acclimation negatively impacts the contractility of steelhead trout (Oncorhynchus mykiss) spongy myocardium.

Cardiac stroke volume (SV) is compromised in Atlantic cod and rainbow trout following acclimation to hypoxia (i.e., 40% air saturation; ~8 kPa O2) at 10-12°C, and this is not due to changes in heart morphometrics or maximum achievable in vitro end-diastolic volume. To examine if this diminished SV may be related to compromised myocardial contractility, we used the work-loop method to measure work and power in spongy myocardial strips from normoxic- and hypoxic-acclimated steelhead trout when exposed to decreasing Po2 levels (21 to 1.5 kPa) at several frequencies (30-90 contractions/min) at 14°C (their acclimation temperature). Work required to lengthen the muscle, as during filling of the heart, was strongly frequency dependent (i.e., increased with contraction rate) but was not affected by hypoxic acclimation or test Po2. In contrast, although shortening work was less frequency dependent, this parameter and network (and power) 1) were consistently lower (by ~30-50 and ~15%, respectively) in strips from hypoxic-acclimated fish and 2) fell by ~40-50% in both groups from 20 to 1.5 kPa Po2, despite the already-reduced myocardial performance in the hypoxic-acclimated group. In addition, strips from hypoxic-acclimated trout showed a poorer recovery of net power (by ~15%) when returned to normoxia. These results strongly suggest that hypoxic acclimation reduces myocardial contractility, and in turn, may limit SV (possibly by increasing end-systolic volume), but that this diminished performance does not improve the capacity to maintain myocardial performance under oxygen limiting conditions.

Knockdown of glycine decarboxylase complex alters photorespiratory carbon isotope fractionation in Oryza sativa leaves.

The influence of reduced glycine decarboxylase complex (GDC) activity on leaf atmosphere CO2 and 13CO2 exchange was tested in transgenic Oryza sativa with the GDC H-subunit knocked down in leaf mesophyll cells. Leaf measurements on transgenic gdch knockdown and wild-type plants were carried out in the light under photorespiratory and low photorespiratory conditions (i.e. 18.4 kPa and 1.84 kPa atmospheric O2 partial pressure, respectively), and in the dark. Under approximately current ambient O2 partial pressure (18.4 kPa pO2), the gdch knockdown plants showed an expected photorespiratory-deficient phenotype, with lower leaf net CO2 assimilation rates (A) than the wild-type. Additionally, under these conditions, the gdch knockdown plants had greater leaf net discrimination against 13CO2 (Δo) than the wild-type. This difference in Δo was in part due to lower 13C photorespiratory fractionation (f) ascribed to alternative decarboxylation of photorespiratory intermediates. Furthermore, the leaf dark respiration rate (Rd) was enhanced and the 13CO2 composition of respired CO2 (δ13CRd) showed a tendency to be more depleted in the gdch knockdown plants. These changes in Rd and δ13CRd were due to the amount and carbon isotopic composition of substrates available for dark respiration. These results demonstrate that impairment of the photorespiratory pathway affects leaf 13CO2 exchange, particularly the 13C decarboxylation fractionation associated with photorespiration.

Hypoxically Induced Nitric Oxide: Potential Role as a Vasodilator in Mytilus edulis Gills.

Intertidal Mytilus edulis experience rapid transgression to hypoxia when they close their valves during low tide. This induces a physiological stress response aiming to stabilize tissue perfusion against declining oxygen partial pressure in shell water. We hypothesized that nitric oxide (NO) accumulation supports blood vessel opening in hypoxia and used live imaging techniques to measure NO and superoxide anion () formation in hypoxia-exposed gill filaments. Thirty minutes of moderate (7 kPa pO2) and severe hypoxia (1 kPa pO2) caused 1.6- and 2.4-fold increase, respectively, of NO accumulation in the endothelial muscle cells of the hemolymphatic vessels of the gill filaments. This led to a dilatation of blood vessel diameter by 43% (7 kPa) and 56% (1 kPa), which facilitates blood flow. Experiments in which we applied the chemical NO-donor Spermine NONOate (concentrations ranging from 1 to 6 mM) under normoxic conditions corroborate the dilatational effect of NO on the blood vessel. The formation of within the filament epithelial cells increased 1.5 (7 kPa) and 2-fold (1 kPa) upon treatment. Biochemical analysis of mitochondrial electron transport complexes in hypoxia-exposed gill tissue indicates decreased activity of complexes I and III in both hypoxic conditions; whereas complex IV (cytochrome-c oxidase) activity increased at 7 kPa and decreased at 1 kPa compared to normoxic exposure conditions. This corresponds to the pattern of pO2-dependent gill respiration rates recorded in ex-vivo experiments. Severe hypoxia (1 kPa) appears to have a stabilizing effect on NO accumulation in gill cells, since less O2 is available for NO oxidation to nitrite/nitrate. Hypoxia thus supports the NO dependent inhibition of complex IV activity, a mechanism that could fine tune mitochondrial respiration to the local O2 availability in a tissue. Our study highlights a basal function of NO in improving perfusion of hypoxic invertebrate tissues, which could be a key mechanism of tolerance toward environmental O2 variations.

Diel cycling hypoxia enhances hypoxia tolerance in rainbow trout (Oncorhynchus mykiss): evidence of physiological and metabolic plasticity.

Many fish naturally encounter a daily cycle of hypoxia, but it is unclear whether this exposure hardens hypoxia-intolerant fish to future hypoxia or leads to accumulated stress and death. The rainbow trout (Oncorhynchus mykiss) is a putatively hypoxia-sensitive species found in rivers and estuaries that may routinely experience hypoxic events. Trout were exposed to one of four 135 h treatments in a swim-tunnel respirometer: (1) air-saturated control (20.7 kPa PO2 ); (2) diel cycling O2 (20.7-4.2 kPa PO2 over 24 h); (3) acute hypoxia (130 h at 20.7 kPa PO2 followed by 5 h at 4.2 kPa PO2 ); and (4) the mean oxygen tension (12.4 kPa PO2 ) experienced by the diel cycled fish. Some responses were similar in diel O2 cycled and mean PO2 -treated fish, but overall, exposure to ecologically representative diel hypoxia cycles improved hypoxia tolerance. Diel hypoxia-induced protective responses included increased inducible HSP70 concentration and mean corpuscular hemoglobin concentration, as well as reduced plasma cortisol. Acclimation to diel hypoxia allowed metabolic rates to decline during hypoxia, reduced oxygen debt following subsequent exposures, and allowed fish to return to an anabolic phenotype. The data demonstrate that acute diel cycling hypoxia improves hypoxia tolerance in previously intolerant fish through the activation of cellular protective mechanisms and a reduction in metabolic O2 requirements.

Mathematical arterialisation of peripheral venous blood gas for obtainment of arterial blood gas values: a methodological validation study in the clinical setting.

Arterial blood gas (ABG) analysis is an essential tool in the clinical assessment of acutely ill patients. Venous to arterial conversion (v-TAC), a mathematical method, has been developed recently to convert peripheral venous blood gas (VBG) values to arterialized VBG (aVBG) values. The aim of this study was to test the validity of aVBG compared to ABG in an emergency department (ED) setting. Twenty ED patients were included in this study. ABG and three aVBG samples were collected from each patient. The aVBG samples were processed in three different ways to investigate appropriate sample handling. All VBG samples were arterialized using the v-TAC method. ABG and aVBG samples were compared using Lin's concordance correlation coefficient (CCC), Bland-Altman plots and misclassification analysis. Clinical acceptable threshold of aVBG value deviance from ABG values were ± 0.05 pH units, ± 0.88kPa pCO2 and ± 0.88kPa pO2. CCC revealed an agreement in pH and pCO2 parameters for both aVBG in comparison to ABG. In all aVBG samples, an overestimation of pO2 compared to ABG was observed. Bland-Altman plot revealed clinically acceptable mean difference and limits-of-agreement intervals between ABG and aVBG pH and pCO2, but not between ABG and aVBG pO2. Arterialization of VBG using v-TAC is a valid method for measuring pH and pCO2, but not for pO2. Larger clinical studies are required to evaluate the applicability of v-TAC in different patient subpopulations.

American alligator (Alligator mississippiensis) embryos tightly regulate intracellular pH during a severe acidosis

Crocodilian nests naturally experience high CO2(hypercarbia), which leads to increased blood Pco2and reduced blood pH (pHe) in embryos; their response to acid–base challenges is not known. During acute hypercarbia, snapping turtle embryos preferentially regulate tissue pH (pHi) against pHereductions. This is proposed to be associated with CO2tolerance in reptilian embryos and is not found in adults. In the present study, we investigated pH regulation in American alligator (Alligator mississippiensis (Daudin, 1802)) embryos exposed to 1 h of hypercarbia hypoxia (13 kPa Pco2, 9 kPa Po2). Hypercarbia hypoxia reduced pHeby 0.42 pH unit, while heart and brain pHiincreased, with no change in the pHiof other tissues. The results indicate that American alligator embryos preferentially regulate pHi, similar to snapping turtle embryos, which represents a markedly different strategy of acid–base regulation than what is observed in adult reptiles. These findings suggest that preferential pHiregulation may be a strategy of acid–base regulation used by embryonic reptiles.

Hypersalinity as a trigger of seagrass (Thalassia testudinum) die-off events in Florida Bay: Evidence based on shoot meristem O2 and H2S dynamics

The mechanisms initiating rapid, large-scale (>50 km2) seagrass die-off events globally remain elusive. Thalassia testudinum, a dominant habitat-forming tropical seagrass of the Caribbean and tropical Atlantic region, experiences recurrent die-off events in Florida Bay. Thus, T. testudinum provides an excellent case study to identify triggers of large-scale seagrass mortality. Presently, seagrass die-off events in Florida Bay are correlated to high total sulfide (H2S, HS−, S2−) concentrations in the sediment porewater (≥2000 μM), water column hypoxia, high temperature and hypersalinity (salinity > 45). Because the mortality appears to be initiated at the shoot base within the meristem, we examined the response of shoot meristematic O2 and H2S dynamics using microsensors over a 5-hour nighttime simulation. Shoots were held at salinity 35 (ambient) or 65 (hypersaline) using microsensors in intact cores with plants from the field. The rate of H2S intrusion in the dark and oxidation in the light were similar at 35 and 65 salinity. However, the tissue O2 consumption rate was significantly higher under hypersaline conditions (−11.07 ± 4.32 kPa pO2 h−1) compared to ambient (−3.93 ± 0.88 kPa pO2 h−1) in the dark. Consequently, the meristematic pO2 threshold (~1.5 kPa O2) where H2S intrusion occurred in the meristem was reached more rapidly, increasing the time H2S accumulated (1.5 to 2.8 h). Longer H2S accumulation time significantly increased maximum meristem H2S levels at 65 (536 ± 330 μM H2S) compared to 35 salinity (121 ± 62 μM H2S). These results in T. testudinum provide evidence for a triggering mechanism that links an enhanced respiratory rate under hypersaline conditions to sulfide toxicity. We propose that hypersalinity in Florida Bay, or any stressor that significantly increases nighttime respiration rates, can subject seagrasses to longer and higher concentrations of sulfide, a known phytotoxin, within the shoot meristem. This mechanism likely explains large-scale mass mortality of T. testudinum in Florida Bay during periods of high salinity and elevated porewater sulfide levels, although field experiments are required to further validate this supposition.

Hypoxia Tolerance and Metabolic Suppression in Oxygen Minimum Zone Euphausiids: Implications for Ocean Deoxygenation and Biogeochemical Cycles.

The effects of regional variations in oxygen and temperature levels with depth were assessed for the metabolism and hypoxia tolerance of dominant euphausiid species. The physiological strategies employed by these species facilitate prediction of changing vertical distributions with expanding oxygen minimum zones and inform estimates of the contribution of vertically migrating species to biogeochemical cycles. The migrating species from the Eastern Tropical Pacific (ETP), Euphausia eximia and Nematoscelis gracilis, tolerate a Partial Pressure (PO2) of 0.8 kPa at 10 °C (∼15 µM O2) for at least 12 h without mortality, while the California Current species, Nematoscelis difficilis, is incapable of surviving even 2.4 kPa PO2 (∼32 µM O2) for more than 3 h at that temperature. Euphausia diomedeae from the Red Sea migrates into an intermediate oxygen minimum zone, but one in which the temperature at depth remains near 22 °C. Euphausia diomedeae survived 1.6 kPa PO2 (∼22 µM O2) at 22 °C for the duration of six hour respiration experiments. Critical oxygen partial pressures were estimated for each species, and, for E. eximia, measured via oxygen consumption (2.1 kPa, 10 °C, n = 2) and lactate accumulation (1.1 kPa, 10 °C). A primary mechanism facilitating low oxygen tolerance is an ability to dramatically reduce energy expenditure during daytime forays into low oxygen waters. The ETP and Red Sea species reduced aerobic metabolism by more than 50% during exposure to hypoxia. Anaerobic glycolytic energy production, as indicated by whole-animal lactate accumulation, contributed only modestly to the energy deficit. Thus, the total metabolic rate was suppressed by ∼49-64%. Metabolic suppression during diel migrations to depth reduces the metabolic contribution of these species to vertical carbon and nitrogen flux (i.e., the biological pump) by an equivalent amount. Growing evidence suggests that metabolic suppression is a widespread strategy among migrating zooplankton in oxygen minimum zones and may have important implications for the economy and ecology of the oceans. The interacting effects of oxygen and temperature on the metabolism of oceanic species facilitate predictions of changing vertical distribution with climate change.

Temperature and the Ventilatory Response to Hypoxia in Gromphadorhina portentosa (Blattodea: Blaberidae).

In general, insects respond to hypoxia by increasing ventilation frequency, as seen in most other animals. Higher body temperatures usually also increase ventilation rates, likely due to increases in metabolic rates. In ectothermic air-breathing vertebrates, body temperatures and hypoxia tend to interact significantly, with an increasing responsiveness of ventilation to hypoxia at higher temperatures. Here, we tested whether the same is true in insects, using the Madagascar hissing cockroach, Gromphadorhina portentosa (Schaum) (Blattodea: Blaberidae). We equilibrated individuals to a temperature (beginning at 20 °C), and animals were exposed to step-wise decreases in PO2 (21, 15, 10, and 5 kPa, in that order), and we measured ventilation frequencies from videotapes of abdominal pumping after 15 min of exposure to the test oxygen level. We then raised the temperature by 5 °C, and the protocol was repeated, with tests run at 20, 25, 30, and 35 °C. The 20 °C animals had high initial ventilation rates, possibly due to handling stress, so these animals were excluded from subsequent analyses. Across all temperatures, ventilation increased in hypoxia, but only significantly at 5 kPa PO2 Surprisingly, there was no significant interaction between temperature and oxygen, and no significant effect of temperature on ventilation frequency from 25 to 35 °C. Plausibly, the rise in metabolic rates at higher temperatures in insects is made possible by increasing other aspects of gas exchange, such as decreasing internal PO2, or increases in tidal volume, spiracular opening (duration or amount), or removal of fluid from the tracheoles.

Copper(I) complexes as alternatives to iridium(III) complexes for highly efficient oxygen sensing.

The complex [Cu(xantphos)(dmp)][PF6] (dmp = 2,9-dimethyl-1,10-phenanthroline) in a nanostructured metal oxyde matrix shows better sensitivity to oxygen (KSV = 9.74 ± 0.87 kPa(-1) between 0 and 1 kPa pO2 and 5.59 ± 0.15 kPa(-1) between 0 and 10 kPa pO2) than cyclometallated iridium complexes in the same conditions.

Hypobaria and hypoxia affects phytochemical production, gas exchange, and growth of lettuce

Hypobaria (low total atmospheric pressure) is essential in sustainable, energy-efficient plant production systems for long-term space exploration and human habitation on the Moon and Mars. There are also critical engineering, safety, and materials handling advantages of growing plants under hypobaria, including reduced atmospheric leakage from extraterrestrial base environments. The potential for producing crops under hypobaria and manipulating hypoxia (low oxygen stress) to increase health-promoting bioactive compounds is not well characterized. Here we showed that hypobaric-grown lettuce plants (25 kPa ≈ 25% of normal pressure) exposed to hypoxia (6 kPa pO2 ≈ 29% of normal pO2) during the final 3 d of the production cycle had enhanced antioxidant activity, increased synthesis of anthocyananins, phenolics, and carotenoids without reduction of photosynthesis or plant biomass. Net photosynthetic rate (P N) was not affected by total pressure. However, 10 d of hypoxia reduced P N, dark respiration rate (R D), P N/R D ratio, and plant biomass. Growing plants under hypobaria and manipulating hypoxia during crop production to enhance health-promoting bioactive compounds is important for the health and well-being of astronauts exposed to space radiation and other stresses during long-term habitation.

The physiological response of the marine platyhelminth Macrostomum lignano to different environmental oxygen concentrations

The respiration rate of meiofauna is difficult to measure, and the response to variations in the environmental oxygen concentration has so far been mainly addressed through behavioral investigation. We investigated the effect of different oxygen concentrations on the physiology of the marine platyhelminth Macrostomum lignano. Respiration was measured using batches of 20 animals in a glass microtiter plate equipped with optical oxygen sensor spots. At higher oxygen saturations (>12 kPa), the animals showed a clear oxyconforming behavior. However, below this value, the flatworms kept respiration rates constant at 0.064±0.001 nmol O2 l(-1) h(-1) individual(-1) down to 3 kPa PO2, and this rate was increased by 30% in animals that were reoxygenated after enduring a period of 1.5 h in anoxia. Physiological changes related to tissue oxygenation were assessed using live imaging techniques with different fluorophores in animals maintained in normoxic (21 kPa), hyperoxic (40 kPa) or near-anoxic (~0 kPa) conditions and subjected to anoxia-reoxygenation. The pH-sensitive dyes Ageladine-A and BCECF both indicated that pHi under near-anoxia increases by about 0.07-0.10 units. Mitochondrial membrane potential, Δψm, was higher in anoxic and hyperoxic than in normoxic conditions (JC1 dye data). Staining with ROS-sensitive dyes - DHE for detection of superoxide anion (O2•(-)) formation and C-H DFFDA for other ROS species aside from O2•(-) (H2O2, HOO• and ONOO) - showed increased ROS formation following anoxia-reoxygenation treatment. Animals exposed to hyperoxic, normoxic and anoxic treatments displayed no significant differences in O2•(-) formation, whereas mitochondrial ROS formation as detected by C-H2DFFDA was higher after hyperoxic exposure and lowest under near-anoxia conditions compared with the normoxic control group. Macrostomum lignano seems to be a species that is tolerant of a wide range of oxygen concentrations (being able to maintain aerobic metabolism from extremely low PO2 up to hyperoxic conditions), which is an essential prerequisite for successfully dealing with the drastic environmental oxygen variations that occur within intertidal sediments.

Early genetic responses in rat vascular tissue after simulated diving

Diving causes a transient reduction of vascular function, but the mechanisms behind this are largely unknown. The aim of this study was therefore to analyze genetic reactions that may be involved in acute changes of vascular function in divers. Rats were exposed to 709 kPa of hyperbaric air (149 kPa Po(2)) for 50 min followed by postdive monitoring of vascular bubble formation and full genome microarray analysis of the aorta from diving rats (n = 8) and unexposed controls (n = 9). Upregulation of 23 genes was observed 1 h after simulated diving. The differential gene expression was characteristic of cellular responses to oxidative stress, with functions of upregulated genes including activation and fine-tuning of stress-responsive transcription, cytokine/cytokine receptor signaling, molecular chaperoning, and coagulation. By qRT-PCR, we verified increased transcription of neuron-derived orphan receptor-1 (Nr4a3), plasminogen activator inhibitor 1 (Serpine1), cytokine TWEAK receptor FN14 (Tnfrsf12a), transcription factor class E basic helix-loop-helix protein 40 (Bhlhe40), and adrenomedullin (Adm). Hypoxia-inducible transcription factor HIF1 subunit HIF1-α was stabilized in the aorta 1 h after diving, and after 4 h there was a fivefold increase in total protein levels of the procoagulant plasminogen activator inhibitor 1 (PAI1) in blood plasma from diving rats. The study did not have sufficient power for individual assessment of effects of hyperoxia and decompression-induced bubbles on postdive gene expression. However, differential gene expression in rats without venous bubbles was similar to that of all the diving rats, indicating that elevated Po(2) instigated the observed genetic reactions.

Allometric scaling of discontinuous gas exchange patterns in the locust Locusta migratoria throughout ontogeny

The discontinuous gas exchange cycle (DGC) is a three-phase breathing pattern displayed by many insects at rest. The pattern consists of an extended breath-hold period (closed phase), followed by a sequence of rapid gas exchange pulses (flutter phase), and then a period in which respiratory gases move freely between insect and environment (open phase). This study measured CO(2) emission in resting locusts Locusta migratoria throughout ontogeny, in normoxia (21 kPa P(O2)), hypoxia (7 kPa P(O2)) and hyperoxia (40 kPa P(O2)), to determine whether body mass and ambient O(2) affect DGC phase duration. In normoxia, mean CO(2) production rate scales with body mass (M(b); g) according to the allometric power equation , closed phase duration (C; min) scales with body mass according to the equation C=8.0M(b)(0.38±0.29), closed+flutter period (C+F; min) scales with body mass according to the equation C+F=26.6M (0.20±0.25)(b) and open phase duration (O; min) scales with body mass according to the equation O=13.3M(b) (0.23±0.18). Hypoxia results in a shorter C phase and longer O phase across all life stages, whereas hyperoxia elicits shorter C, C+F and O phases across all life stages. The tendency for larger locusts to exhibit longer C and C+F phases might arise if the positive allometric scaling of locust tracheal volume prolongs the time taken to reach the minimum O(2) and maximum CO(2) set-points that determine the duration of these respective periods, whereas an increasingly protracted O phase could reflect the additional time required for larger locusts to expel CO(2) through a relatively longer tracheal pathway. Observed changes in phase duration under hypoxia possibly serve to maximise O(2) uptake from the environment, whereas the response of the DGC to hyperoxia is difficult to explain, but could be affected by elevated levels of reactive oxygen species.

Low O2 acclimation shifts the hypoxia avoidance behaviour of snapper (Pagrus auratus) with only subtle changes in aerobic and anaerobic function

It was hypothesised that chronic hypoxia acclimation (preconditioning) would alter the behavioural low-O(2) avoidance strategy of fish as a result of both aerobic and anaerobic physiological adaptations. Avoidance and physiological responses of juvenile snapper (Pagrus auratus) were therefore investigated following a 6 week period of moderate hypoxia exposure (10.2-12.1 kPa P(O(2)), 21 ± 1 °C) and compared with those of normoxic controls (P(O(2))=20-21 kPa, 21 ± 1 °C). The critical oxygen pressure (P(crit)) limit of both groups was unchanged at ~7 kPa, as were standard, routine and maximum metabolic rates. However, hypoxia-acclimated fish showed increased tolerances to hypoxia in behavioural choice chambers by avoiding lower P(O(2)) levels (3.3 ± 0.7 vs 5.3 ± 1.1 kPa) without displaying greater perturbations of lactate or glucose. This behavioural change was associated with unexpected physiological adjustments. For example, a decrease in blood O(2) carrying capacity was observed after hypoxia acclimation. Also unexpected was an increase in whole-blood P(50) following acclimation to low O(2), perhaps facilitating Hb-O(2) off-loading to tissues. In addition, cardiac mitochondria measured in situ using permeabilised fibres showed improved O(2) uptake efficiencies. The proportion of the anaerobic enzyme lactate dehydrogenase, at least relative to the aerobic marker enzyme citrate synthase, also increased in heart and skeletal red muscle, indicating enhanced anaerobic potential, or in situ lactate metabolism, in these tissues. Overall, these data suggest that a prioritization of O(2) delivery and O(2) utilisation over O(2) uptake during long-term hypoxia may convey a significant survival benefit to snapper in terms of behavioural low-O(2) tolerance.

Evaluation of a mathematical model of the acid‐base chemistry of blood

The objective of this study is to evaluate whether a mathematical model of acid‐base chemistry of blood [1,2] can simulate physiological situations such as the mixing of blood with different acid‐base status. To do so, two samples of venous blood from 21 normal subjects and 6 patients were equilibrated at different O2 and CO2 partial pressures and then mixed. The mathematical model was used to simulate this mixing and model‐simulated and measured values compared. Model simulations compared well with measured values with a small bias and standard deviation, within normal analytical variability, e.g. for normal subjects 0.000±0.006 pH, −0.06±0.11 kPa PCO2, −0.1±0.8 % SO2 and −0.02±0.13 kPa PO2. This model is the first based on a mass‐balance, mass‐action formulation which can perform simulations of this type, including both hemoglobin buffering and the competitive binding of O2, CO2 and H+ on hemoglobin.