Low O2 Conditions Research Articles

Characterization of denitrification and net N2O-reduction properties of novel aerobically N2O-reducing bacteria

ABSTRACTNitrous oxide, one of the earth-warming and ozone-destructing gases, is produced through either nitrification or denitrification depending on the O2 availability in soil. Aerobically denitrifying bacteria express denitrification tract even under the gas phase containing O2 at the ambient air level. The net reduction of exogenous N2O by novel aerobically denitrifying bacteria were studied. We carried out two different isolation strategies in the primary screening. One was to select isolates of interest out of periplasmic nitrate reductase-dependent denitrifying bacteria in a eutrophic condition. The other was to use diluted nutrient agar to allow the formation of colonies of diverse bacteria. Among aerobically denitrifying bacteria, those which showed net aerobic N2O reduction were only minor populations. As a result, eight isolates belonging to Proteobacteria were obtained from soil and cow manure. The denitrification and net N2O reduction properties of the three representative isolates, Pseudomonas sp. CM1, Thauera sp. PM2 and Paracoccus denitrificans 96, were determined separately by the acetylene inhibition method after exposure to aerobic or low O2 conditions, a 24 h starvation prior to the determination of the aerobic activity and inoculation to a cow manure-amended sterile soil. The phenotype inversion from net N2O-reducing to N2O-emitting, and vice versa, attested to the fact that activity of the N2O-producing and -reducing steps changed in different intensities to each other. The activity values and the direction of activity changes varied among the isolates. When they were inoculated in a sterilized soil microcosm at 40% maximum water holding capacity, the denitrification and the N2O-reducing activities were comparable with or, in some cases, facilitated more than those determined under the low-O2 condition. It is possible that these isolates sensed the O2 deficiency even in such a relatively dry condition. Pseudomonas sp. CM1 was unique because it lacked nitrate reducing activity and acted as a net aerobic N2O reducer.

Discovery and dissection of metabolic oscillations in the microaerobic nitric oxide response network of Escherichia coli

The virulence of many pathogens depends upon their ability to cope with immune-generated nitric oxide (NO·). In Escherichia coli, the major NO· detoxification systems are Hmp, an NO· dioxygenase (NOD), and NorV, an NO· reductase (NOR). It is well established that Hmp is the dominant system under aerobic conditions, whereas NorV dominates anaerobic conditions; however, the quantitative contributions of these systems under the physiologically relevant microaerobic regime remain ill defined. Here, we investigated NO· detoxification in environments ranging from 0 to 50 μM O2, and discovered a regime in which E. coli NO· defenses were severely compromised, as well as conditions that exhibited oscillations in the concentration of NO·. Using an integrated computational and experimental approach, E. coli NO· detoxification was found to be extremely impaired at low O2 due to a combination of its inhibitory effects on NorV, Hmp, and translational activities, whereas oscillations were found to result from a kinetic competition for O2 between Hmp and respiratory cytochromes. Because at least 777 different bacterial species contain the genetic requirements of this stress response oscillator, we hypothesize that such oscillatory behavior could be a widespread phenomenon. In support of this hypothesis,Pseudomonas aeruginosa, whose respiratory and NO· response networks differ considerably from those of E. coli, was found to exhibit analogous oscillations in low O2 environments. This work provides insight into how bacterial NO· defenses function under the low O2 conditions that are likely to be encountered within host environments.

Oxygen dependency of germinating Brassica seeds

Establishing plants in space, Moon or Mars requires adaptation to altered conditions, including reduced pressure and composition of atmospheres. To determine the oxygen requirements for seed germination, we imbibed Brassica rapa seeds under varying oxygen concentrations and profiled the transcription patterns of genes related to early metabolism such as starch degradation, glycolysis, and fermentation. We also analyzed the activity of lactate dehydrogenase (LDH) and alcohol dehydrogenase (ADH), and measured starch degradation. Partial oxygen pressure (pO2) greater than 10% resulted in normal germination (i.e., protrusion of radicle about 18 hours after imbibition) but lower pO2 delayed and reduced germination. Imbibition in an oxygen-free atmosphere for three days resulted in no germination but subsequent transfer to air initiated germination in 75% of the seeds and the root growth rate was transiently greater than in roots germinated under ambient pO2. In hypoxic seeds soluble sugars degraded faster but the content of starch after 24 h was higher than at ambient oxygen. Transcription of genes related to starch degradation, α-amylase (AMY) and Sucrose Synthase (SUS), was higher under ambient O2 than under hypoxia. Glycolysis and fermentation pathway-related genes, glucose phosphate isomerase (GPI), 6-phosphofructokinase (PFK), fructose 1,6-bisphosphate aldolase (ALD), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), pyruvate decarboxylase (PDC), LDH, and ADH, were induced by low pO2. The activity of LDH and ADH was the highest in anoxic seeds. Germination under low O2 conditions initiated ethanolic fermentation. Therefore, sufficient oxygen availability is important for germination before photosynthesis provides necessary oxygen and the determination of an oxygen carrying capacity is important for uniform growth in space conditions.

Three Pseudomonas putida FNR Family Proteins with Different Sensitivities to O2

The Escherichia coli fumarate-nitrate reduction regulator (FNR) protein is the paradigm for bacterial O2-sensing transcription factors. However, unlike E. coli, some bacterial species possess multiple FNR proteins that presumably have evolved to fulfill distinct roles. Here, three FNR proteins (ANR, PP_3233, and PP_3287) from a single bacterial species, Pseudomonas putida KT2440, have been analyzed. Under anaerobic conditions, all three proteins had spectral properties resembling those of [4Fe-4S] proteins. The reactivity of the ANR [4Fe-4S] cluster with O2 was similar to that of E. coli FNR, and during conversion to the apo-protein, via a [2Fe-2S] intermediate, cluster sulfur was retained. Like ANR, reconstituted PP_3233 and PP_3287 were converted to [2Fe-2S] forms when exposed to O2, but their [4Fe-4S] clusters reacted more slowly. Transcription from an FNR-dependent promoter with a consensus FNR-binding site in P. putida and E. coli strains expressing only one FNR protein was consistent with the in vitro responses to O2. Taken together, the experimental results suggest that the local environments of the iron-sulfur clusters in the different P. putida FNR proteins influence their reactivity with O2, such that ANR resembles E. coli FNR and is highly responsive to low concentrations of O2, whereas PP_3233 and PP_3287 have evolved to be less sensitive to O2.

Involvement of hepatic stellate cell cytoglobin in acute hepatocyte damage through the regulation of CYP2E1-mediated xenobiotic metabolism

Oxygen (O2) is required for cytochrome P450 (CYP)-dependent drug metabolism. Cytoglobin (CYGB) is a unique globin expressed exclusively in hepatic stellate cells (HSCs). However, its role in O2-dependent metabolism in neighboring hepatocytes remains unknown. This study provides evidence that CYGB in HSCs is involved in acetaminophen (N-acetyl-p-aminophenol; APAP)-induced hepatotoxicity. Serum alanine aminotransferase levels were higher in wild-type mice than in Cygb-null mice. Wild-type mice exhibited more severe hepatocyte necrosis around the central vein area compared with Cygb-null mice, thus indicating that CYGB deficiency protects against APAP-induced liver damage. Although no difference in the hepatic expression of CYP2E1, a key enzyme involved in APAP toxicity, was observed between wild-type and Cygb-null mice, the serum levels of the APAP metabolites cysteinyl-APAP and N-acetyl-cysteinyl-APAP were decreased in Cygb-null mice, suggesting reduced APAP metabolism in the livers of Cygb-null mice. In primary cultures, APAP-induced hepatocyte damage was increased by co-culturing with wild-type HSCs but not with Cygb-null HSCs. In addition, cell damage was markedly alleviated under low O2 condition (5% O2), suggesting the requirement of O2 for APAP toxicity. Carbon tetrachloride-induced liver injury (CYP2E1-dependent), but not lipopolysaccharide/D-galactosamine-induced injury (CYP2E1-independent), was similarly alleviated in Cygb-null mice. Considering the function of CYGB as O2 carrier, these results strongly support the hypothesis that HSCs are involved in the CYP2E1-mediated xenobiotic activation by augmenting O2 supply to hepatocytes. In conclusion, CYGB in HSCs contributes to the CYP-mediated metabolism of xenobiotics in hepatocytes by supplying O2 for enzymatic oxidation.

Refuse-Derived Fuel and Integrated Calcium Hydroxide Sorbent for Coal Combustion Desulfurization

Pelletized refuse-derived fuel (RDF; 65 wt % nonrecyclable fibers/plastics) and integrated calcium hydroxide sorbent (35 wt %), referred to as a ReEngineered Feedstock (ReEF), were evaluated as a renewable fuel and an in situ sulfur sorbent within pulverized coal combustion. Characterization of ReEF with scanning electron microscopy–energy-dispersive X-ray spectroscopy indicated random distribution of calcium hydroxide sorbents on fiber/plastic particles. The sulfation kinetics of ReEF combustion were evaluated in a drop-tube reactor at variable conditions (800–1000 °C, 1400 ppm of SO2, and 5–30% O2 in N2) with continuous effluent characterization via online IR spectroscopy. Sulfation of calcium hydroxide in ReEF was delayed because of RDF combustion when compared with pure calcium hydroxide sorbent. A low-O2 condition was shown to produce high conversion of calcium hydroxide to calcium sulfate (XCa(OH)2 > 0.4), while calcination and combustion of ReEF in a high-O2 environment (O2 > 20%) resulted in incre...

Effect of carbon dioxide on the high temperature transformation of siderite under low oxygen conditions

The knowledge of siderite transformation is critical to the understanding of ash deposition behavior in oxy-fuel combustion. This work aims to explore the effect of CO2 on the transformation of siderite in different modes under low O2 conditions, which are common in real boilers. Two samples, i.e. pure siderite and excluded siderite simulated by the addition of siderite into a demineralized coal sample, were combusted at 1350°C in a drop tube furnace. Two conditions, i.e. O2/N2=7/93 and O2/CO2=7/93 were tested for elucidating the effect of CO2. The combustion products were analyzed by scanning electron microscopy and X-ray diffraction. The results show that, for pure siderite, the products generated in both conditions have similar morphology and mineralogy, suggesting negligible effects of CO2. They are dominated by irregular particles with sharp edges, and only magnetite is detected. However, for excluded siderite, the products generated in both conditions are distinctly different in morphology and mineralogy, which indicates significant effects of CO2 on the transformation of excluded siderite. The product generated in O2/CO2=7/93 is dominated by irregular particles with rounded edges, while that generated in O2/N2=7/93 is dominated by spherical particles. Wustite and magnetite are detected in both products, but the production of magnetite is much higher in O2/CO2=7/93 than that in O2/N2=7/93, which is attributed to the presence of a high concentration of CO2. Compared with pure siderite, excluded siderite experiences more reducing conditions and higher temperatures due to coal combustion, which results in the survival of wustite and more extensive melting.

Flood adaptive traits and processes: an overview.

Unanticipated flooding challenges plant growth and fitness in natural and agricultural ecosystems. Here we describe mechanisms of developmental plasticity and metabolic modulation that underpin adaptive traits and acclimation responses to waterlogging of root systems and submergence of aerial tissues. This includes insights into processes that enhance ventilation of submerged organs. At the intersection between metabolism and growth, submergence survival strategies have evolved involving an ethylene-driven and gibberellin-enhanced module that regulates growth of submerged organs. Opposing regulation of this pathway is facilitated by a subgroup of ethylene-response transcription factors (ERFs), which include members that require low O₂ or low nitric oxide (NO) conditions for their stabilization. These transcription factors control genes encoding enzymes required for anaerobic metabolism as well as proteins that fine-tune their function in transcription and turnover. Other mechanisms that control metabolism and growth at seed, seedling and mature stages under flooding conditions are reviewed, as well as findings demonstrating that true endurance of submergence includes an ability to restore growth following the deluge. Finally, we highlight molecular insights obtained from natural variation of domesticated and wild species that occupy different hydrological niches, emphasizing the value of understanding natural flooding survival strategies in efforts to stabilize crop yields in flood-prone environments.

Hypoxia-Induced Signaling Promotes Prostate Cancer Progression: Exosomes Role as Messenger of Hypoxic Response in Tumor Microenvironment.

Prostate cancer (PCA) is the leading malignancy in men and the second leading cause of cancer-related deaths. Hypoxia (low O2 condition) is considered an early event in prostate carcinogenesis associated with an aggressive phenotype. In fact, clinically, hypoxia and hypoxia-related biomarkers are associated with treatment failure and disease progression. Hypoxia-inducible factor 1 (HIF-1) is the key factor that is activated under hypoxia, and mediates adaptation of cells to hypoxic conditions through regulating the expression of genes associated with angiogenesis, epithelial-to-mesenchymal transition (EMT), metastasis, survival, proliferation, metabolism, sternness, hormone-refractory progression, and therapeutic resistance. Besides HIF-1, several other signaling pathways including PI3K/Akt/mTOR, NADPH oxidase (NOX), Wnt/b-catenin, and Hedgehog are activated in cancer cells under hypoxic conditions, and also contribute in hypoxia-induced biological effects in HIF-1-dependent and -independent manners. Hypoxic cancer cells cause extensive changes in the tumor microenvironment both local and distant, and recent studies have provided ample evidence supporting the crucial role of nanosized vesicles "exosomes" in mediating hypoxia-induced tumor microenvironment remodeling. Exosomes' role has been reported in hypoxia-induced angiogenesis, sternness, activation of cancer-associated fibroblasts (CAFs), and EMT. Together, existing literature suggests that hypoxia plays a predominant role in PCA growth and progression, and PCA could be effectively prevented and treated via targeting hypoxia/hypoxia-related signaling pathways.

Windrow composting mitigated CH4 emissions: characterization of methanogenic and methanotrophic communities in manure management.

With increasing livestock breeding, methane (CH4 ) emissions from manure management will increasingly contribute more to atmospheric CH4 concentration. The dynamics of methanogens and methanotrophs have not yet been studied in the manure environment. The current study combines surface CH4 emissions with methanogenic and methanotrophic community analyses from two management practices, windrow composting (WCOM) and solid storage (SSTO). Our results showed that there was an c. 50% reduction of CH4 emissions with WCOM compared with SSTO over a 50-day period. A sharp decrease in the quantities of both methanogens and methanotrophs in WCOM suggested that CH4 mitigation was mainly due to decreased CH4 production rather than increased CH4 oxidation. Pyrosequencing analysis demonstrated that aeration caused a clear shift of dominant methanogens in the manure, with specifically a significant decrease in Methanosarcina and increase in Methanobrevibacter. The composition of methanogenic community was influenced by manure management and regulated CH4 production. A sharp increase in the quantity of methanotrophs in SSTO suggested that microbial CH4 oxidation is an important sink for the CH4 produced. The increased abundance of Methylococcaceae in SSTO suggested that Type I methanotrophs have an advantage in CH4 oxidation in occupying niches under low CH4 and high O2 conditions.

Formation of Filamentous Mn Oxide Particles by the Alphaproteobacterium Bosea sp. Strain BIWAKO-01

Mn-rich filamentous particles present in stratified water environments are considered bacteriogenic; however, little is known about their causative agents. This study investigated the production of these particles by an alphaproteobacterium, Bosea sp. strain BIWAKO-01. Particle formation was promoted in static cultures with slightly viscous medium at pH 6.0−6.3 under low-O2 conditions. The Mn(II) oxidation in cultures was slower in higher O2 concentration. These results suggested that pH and O2 concentration are important factors affecting filamentous Mn particle formation in the Mn(II) oxidizer. Lectin staining followed by fluorescence microscopy revealed the presence of specific carbohydrates in the filamentous structures. In addition, transmission electron microscopy, high angle annular dark field scanning transmission electron microscopy, and electron energy loss spectroscopy revealed the structural and spatial associations of Mn with O and C on a nanometer scale in filaments. The results suggested the occurrence of sheet-type Mn oxide likely due to the catalytic activity in exopolymeric substances including acidic polysaccharides.

Deep-water microbialites of the Mesoproterozoic Dismal Lakes Group: microbial growth, lithification, and implications for coniform stromatolites.

Offshore facies of the Mesoproterozoic Sulky Formation, Dismal Lakes Group, arctic Canada, preserve microbialites with unusual morphology. These microbialites grew in water depths greater than several tens of meters and correlate with high-relief conical stromatolites of the more proximal September Lake reef complex. The gross morphology of these microbial facies consists of ridge-like vertical supports draped by concave-upward, subhorizontal elements, resulting in tent-shaped cuspate microbialites with substantial primary void space. Morphological and petrographic analyses suggest a model wherein penecontemporaneous upward growth of ridge elements and development of subhorizontal draping elements initially resulted in a buoyantly supported, unlithified microbial form. Lithification began via precipitation within organic elements during microbialite growth. Mineralization either stabilized or facilitated collapse of initially neutrally buoyant microbialite forms. Microbial structures and breccias were then further stabilized by precipitation of marine herringbone cement. During late-stage diagenesis, remaining void space was occluded by ferroan dolomite cement. Cuspate microbialites are most similar to those found in offshore facies of Neoarchean carbonate platforms and to unlithified, buoyantly supported microbial mats in modern ice-covered Antarctic lakes. We suggest that such unusual microbialite morphologies are a product of the interaction between motile and non-motile communities under nutrient-limiting conditions, followed by early lithification, which served to preserve the resultant microbial form. The presence of marine herringbone cement, commonly associated with high dissolved inorganic carbon (DIC), low O2 conditions, also suggests growth in association with reducing environments at or near the seafloor or in conjunction with a geochemical interface. Predominance of coniform stromatolite forms in the Proterozoic--across a variety of depositional environments--may thus reflect a combination of heterogeneous nutrient distribution, potentially driven by variable redox conditions, and an elevated carbonate saturation state, which permits preservation of these unusual microbialite forms.

High oxygen atmospheres can induce russet spotting development in minimally processed iceberg lettuce

High oxygen modified atmospheres have been suggested as an alternative preservation technique to classical low O2 modified atmosphere packaging (MAP) for fresh-cut lettuce. The advantages of high O2 storage would be a strong reduction of browning, avoidance of low O2 levels linked to off-odors, and the inhibition of microbial growth. However, storage under high O2 potentially could increase both production of ethylene and sensitivity to this hormone in lettuce tissue, leading to the development of quality problems linked to ethylene. In this study, different quality parameters (sensory quality, microbiological load, electrolyte leakage, volatile metabolites) were studied on fresh-cut iceberg lettuce stored under different gas conditions at 7°C: (low O2 MAP (3% O2 compensated with N2), atmospheric conditions, high O2 MAP (50 or 90% O2 compensated with N2)). Furthermore, additional experiments using ethylene absorbers were performed in order to assess the link between high O2 storage, ethylene accumulation, and russet spotting. There was no significant difference between storage conditions regarding growth of mesophilic bacteria and yeasts, electrolyte leakage, or ethanol production. On the other hand, high O2 atmospheres reduced browning but promoted russet spotting development compared with low O2 and atmospheric conditions. In further specific experiments the relationship between high O2 storage, ethylene production, and russet spotting in fresh-cut iceberg lettuce were made evident.

Analysis of the tryptophanase expression in Symbiobacterium thermophilum in a coculture with Geobacillus stearothermophilus.

The tryptophanase-positive Symbiobacterium thermophilum is a free-living syntrophic bacterium that grows effectively in a coculture with Geobacillus stearothermophilus. Our studies have shown that S. thermophilum growth depends on the high CO2 and low O2 condition established by the precedent growth of G. stearothermophilus. The use of an anoxic atmosphere containing high CO2 allows S. thermophilum to grow independently of G. stearothermophilus, but the cellular yield is ten times lower than that achieved in the coculture. In this study, we characterized the coculture-dependent expression and activity of tryptophanase in S. thermophilum. S. thermophilum cells accumulated a marked amount of indole in a coculture with G. stearothermophilus, but not in the bacterium's pure culture irrespective of the addition of tryptophan. S. thermophilum cells accumulated indole in its pure culture consisting of conditioned medium (medium supplied with culture supernatant of G. stearothermophilus). Proteomic analysis identified the protein specifically produced in the S. thermophilum cells grown in conditioned medium, which was a tryptophanase encoded by tna2 (STH439). An attempt to isolate the tryptophanase-inducing component from the culture supernatant of G. stearothermophilus was unsuccessful, but we did discover that the indole accumulation occurs when 10 mM bicarbonate is added to the medium. RT-PCR analysis showed that the addition of bicarbonate stimulated transcription of tna2. The transcriptional start site, identified within the tna2 promoter, was preceded by the -24 and -12 consensus sequences specified by an alternative sigma factor, σ(54). The evidence suggests that the transcription of some genes involved in amino acid metabolism is σ(54)-dependent, and that a bacterial enhancer-binding protein containing a PAS domain controls the transcription under the presence of high levels of bicarbonate.

The response of striped surfperch Embiotoca lateralis to progressive hypoxia: Swimming activity, shoal structure, and estimated metabolic expenditure

When exposed to moderately low O2 conditions (75–30% O2 saturation), shoaling striped surfperch Embiotoca lateralis show no changes in swimming activity, uniform levels of metabolic expenditure and an unchanging shoal structure. As O2 levels become more hypoxic surfperch reduce their locomotor activity, initially with a decrease in turning rate at 20% O2 saturation, followed by a reduction in swimming speed at 15% O2 saturation. Reductions in activity occur in combination with individuals adopting more uniform turning behaviors, which translate into an increase in the distance traveled per unit time (i.e. net displacement). This increase in net displacement may enhance the ability of surfperch to distance themselves from discrete zones of hypoxia. Measures of shoal structure, including nearest neighbor distances and shoal polarity, were constant throughout low O2 exposure despite changes in swimming activity. Estimation of the energetic costs of these behaviors reveals that metabolic expenditure (routine metabolic rate) remained steady at ~120mgO2kg−1h−1 between 100 and 30% O2 saturation, dropping to ~108mgO2kg−1h−1 at 20 and 15% O2 saturation. Given that surfperch possess a critical oxygen saturation (Scrit) of 15.2%, reductions in swimming activity and metabolic energy expenditure clearly occur as individuals reach their aerobic metabolic limits. These results identify that surfperch demonstrate a multi-faceted coping strategy when exposed to low oxygen conditions, which may prove advantageous when the species experience hypoxic episodes in their natural habitat.

Catecholaminergic (CA) neurons modulate hypoxic ventilatory response in newborn rats (P7‐8) (LB737)

CA neurons are involved in autonomic and respiratory regulation during low O2 conditions in adult mammals. We evaluated the participation of central CA neurons of neonate male and female rats (P7‐8) in the hypoxic ventilatory response (HVR) by specifically lesioning them with antidopamine beta‐hydroxylase‐saporin (DBH‐SAP, 42ng / 100nL) injected via the 4th ventricle. Minute ventilation (VE) of newborn was recorded by pressure‐plethysmography from the body chamber during normoxia and hypoxia (10%O2). HVR was reduced significantly (~23%) in male and female lesioned newborn rats (male‐ control group: 137.6±6.4 (% of baseline) vs lesioned group: 104.3±1.4 (% of baseline), p<0.01; female‐ control group: 131±1.0 (% of baseline) vs lesioned group: 102.6±2.6 (% of baseline) p<0.02). These results suggest that catecholaminergic neurons exert an excitatory modulation in newborn rats in O2 chemosensitivity.Grant Funding Source: Fapesp and CNPq

Negative Valve Overlap Reforming Chemistry in Low-Oxygen Environments

Fuel injection into the negative valve overlap (NVO) period is a common method for controlling combustion phasing in homogeneous charge compression ignition (HCCI) and other forms of advanced combustion. When fuel is injected into O2-deficient NVO conditions, a portion of the fuel can be converted to products containing significant levels of H2 and CO. Additionally, other short chain hydrocarbons are produced by means of thermal cracking, water-gas shift, and partial oxidation reactions. The present study experimentally investigates the fuel reforming chemistry that occurs during NVO. To this end, two very different experimental facilities are utilized and their results are compared. One facility is located at Oak Ridge National Laboratory, which uses a custom research engine cycle developed to isolate the NVO event from main combustion, allowing a steady stream of NVO reformate to be exhausted from the engine and chemically analyzed. The other experimental facility, located at Sandia National Laboratories, uses a dump valve to capture the exhaust from a single NVO event for analysis. Results from the two experiments are in excellent trend-wise agreement and indicate that the reforming process under low-O2 conditions produces substantial concentrations of H2, CO, methane, and other short-chain hydrocarbon species. The concentration of thesemore » species is found to be strongly dependent on fuel injection timing and injected fuel type, with weaker dependencies on NVO duration and initial temperature, indicating that NVO reforming is kinetically slow. Further, NVO reforming does not require a large energy input from the engine, meaning that it is not thermodynamically expensive. The implications of these results on HCCI and other forms of combustion are discussed in detail.« less

Human Adipose-Derived Stromal/Stem Cells Induce Functional CD4+CD25+FoxP3+CD127−Regulatory T Cells Under Low Oxygen Culture Conditions

Human adipose tissue stromal/stem cells (ASCs) are known to induce proliferation of resting T cells under ambient (21%) O2 conditions; however, ASCs exist physiologically under lower oxygen (5% O2) conditions in adipose tissue. The effects of low oxygen levels on ASC immunomodulation of T cells are unknown. In this study, we show that ASCs stimulated proliferation of naive CD4(+) T cells and the percentage of CD25(+) T cells was significantly increased under both low and ambient O2. Forkhead box P3 (FoxP3) and transforming growth factor beta (TGF-β) mRNA expression were significantly increased when ASCs were cocultured with CD4(+) T cells under low compared with ambient O2 conditions. The low O2-induced regulatory T cells (iTregs) exhibited functionality when added to mixed lymphocyte reactions as demonstrated by inhibition of peripheral blood mononuclear cell proliferation, and by >300-fold increase in FoxP3 mRNA, and >2-fold increase in TGF-β mRNA expression. These results demonstrate that under physiologically relevant low O2 conditions, direct contact of human ASCs with naive CD4(+) T cells induced functional iTregs.

The effects of intermittent exposure to low-pH and low-oxygen conditions on survival and growth of juvenile red abalone

Abstract. Exposure of nearshore animals to hypoxic, low-pH waters upwelled from below the continental shelf and advected near the coast may be stressful to marine organisms and lead to impaired physiological performance. We mimicked upwelling conditions in the laboratory and tested the effect of fluctuating exposure to water with low-pH and/or low-oxygen levels on the mortality and growth of juvenile red abalone (Haliotis rufescens, shell length 5–10 mm). Mortality rates of juvenile abalone exposed to low-pH (7.5, total scale) and low-O2 (40% saturation, mg L−1) conditions for periods of 3 to 6 h every 3–5 days over 2 weeks did not differ from those exposed to control conditions (O2: 100% saturation, 12 mg L−1; pH 8.0). However, when exposure was extended to 24 h, twice over a 15-day period, juveniles experienced 5–20% higher mortality in the low-oxygen treatments compared to control conditions. Growth rates were reduced significantly when juveniles were exposed to low-oxygen and low-pH treatments. Furthermore, individual variation of growth rate increased when juveniles were exposed simultaneously to low-pH and low-O2 conditions. These results indicate that prolonged exposure to low-oxygen levels is detrimental for the survival of red abalone, whereas pH is a crucial factor for their growth. However, the high individual variation in growth rate under low levels of both pH and oxygen suggests that cryptic phenotypic plasticity may promote resistance to prolonged upwelling conditions by a portion of the population.

Iron Isotope Characteristics of Hot Springs at Chocolate Pots, Yellowstone National Park

Chocolate Pots Hot Springs in Yellowstone National Park is a hydrothermal system that contains high aqueous ferrous iron [∼0.1 mM Fe(II)] at circumneutral pH conditions. This site provides an ideal field environment in which to test our understanding of Fe isotope fractionations derived from laboratory experiments. The Fe(III) oxides, mainly produced through Fe(II) oxidation by oxygen in the atmosphere, have high ⁵⁶Fe/⁵⁴Fe ratios compared with the aqueous Fe(II). However, the degree of fractionation is less than that expected in a closed system at isotopic equilibrium. We suggest two explanations for the observed Fe isotope compositions. One is that light Fe isotopes partition into a sorbed component and precipitate out on the Fe(III) oxide surfaces in the presence of silica. The other explanation is internal regeneration of isotopically heavy Fe(II) via dissimilatory Fe(III) reduction farther down the flow path as well as deeper within the mat materials. These findings provide evidence that silica plays an important role in governing Fe isotope fractionation factors between reduced and oxidized Fe. Under conditions of low ambient oxygen, such as may be found on early Earth or Mars, significantly larger Fe isotope variations are predicted, reflecting the more likely attainment of Fe isotope equilibrium associated with slower oxidation rates under low-O₂ conditions.