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Hypoxia in the root zone activates several mechanisms to cope with the energetic imbalance that triggers hypoxia. Prunus is a hypoxia-sensitive species, and there is not information about the relationship between components of the root respiration and the hypoxia tolerance. We investigated whether adjusting root respiratory components plays a role in the hypoxia tolerance in Prunus rootstocks genotypes. Three Prunus rootstocks genotypes, ‘Mariana 2624’ (tolerant), ‘CAB6P’ (moderately sensitive) and ‘Mazzard F12/1’ (sensitive), were exposed to normoxia and hypoxia in a hydroponic system for 30 days. During the first 10 days of the experiment, the hypoxia-tolerant genotype under hypoxia exhibited a higher root relative growth rate, maintained net nitrogen uptake rate, and had lower protein turnover and electrolyte leakage, as well as increased root porosity (aerenchyma formation from 0.3 to 6.0 cm from the root tip at 30 days of experiment) compared to hypoxia-sensitive genotypes. ‘Mariana 2624’ roots exhibited tolerance to hypoxia by reducing maintenance respiration by 50% compared to normoxia, while growth respiration increased. Control of root respiratory components, early response of ion uptake favors root relative growth rate, and change of the root anatomy are associated with the ability to tolerate hypoxia by reducing maintenance costs. [ABSTRACT FROM AUTHOR]

The article presents community recommendations on terminology and procedures used in flooding and low oxygen stress research. This include flooding, anaerobiosis, anaerobic metabolism, anoxia, hypoxia, hyperoxia/superoxia, stress conditions and scoring survival. It also discusses the challenge of monitoring oxygen levels in plants.

The physiological role of nitrate as a protective factor against anaerobic stress was studied in experiments with tolerant to anoxia sugarcane ( Saccharum officibarum L.) callus lines obtained by in vitro selection in the absence of exogenous carbohydrates. Original cell lines, which were not subjected to selection and therefore more sensitive to oxygen shortage, served as a control. In these lines, anaerobic stress was created in the presence or absence of nitrate in nutrient medium. The presence of nitrate in nutrient medium increased markedly tolerance to anaerobic stress of both lines differing in their sensitivity to anaerobiosis. However, the degree of tolerance differed substantially in compared lines. In the presence of exogenous nitrate, in tolerant cells there were no signs of mitochondrial membrane destruction or degradation even after 72 h of anoxia, whereas in control cells 48-h anaerobic incubation led to the complete degradation of mitochondrial membranes and membranes of other organelles. It is concluded that significant increase in the tolerance of S. officinarum cells in the process of in vitro selection most likely occurred due to induction and stimulation of not only the processes of glycolysis and fermentation, but also nitrate and maybe nitrite utilization. [ABSTRACT FROM AUTHOR]

Cyclic photosynthetic electron flow (CEF) is crucial to photosynthesis because it participates in the control of chloroplast energy and redox metabolism, and it is particularly induced under adverse environmental conditions. Here we report that down-regulation of the chloroplast localized Ca2+ sensor (CAS) protein by an RNAi approach in Chlamydomonas reinhardtii results in strong inhibition of CEF under anoxia. Importantly, this inhibition is rescued by an increase in the extracellular Ca2+ concentration, inferring that CEF is Ca2+-dependent. Furthermore, we identified a protein, anaerobic response 1 (ANR1), that is also required for effective acclimation to anaerobiosis. Depletion of ANR1 by artificial microRNA expression mimics the CAS-depletion phenotype, and under anaerobic conditions the two proteins coexist within a large active photosystem lcytochrome b₆/f complex. Moreover, we provide evidence that CAS and ANR1 interact with each other as well as with PGR5-Like 1 (PGRL1) in vivo. Overall our data establish a Ca2+-dependent regulation of CEF via the combined function of ANR1, CAS, and PGRL1, associated with each other in a multiprotein complex. [ABSTRACT FROM AUTHOR]

Abstract: Lactate dehydrogenase (LDH) is the terminal enzyme of anaerobic glycolysis and key to hypoxia/anoxia survival by most animals. In this study, the effects of anoxic submergence (20h at 7°C in nitrogen-bubbled water) were assessed on LDH from liver of an anoxia-tolerant freshwater turtle, the red-eared slider (Trachemys scripta elegans). Liver LDH from aerobic and anoxic turtles was purified to homogeneity in two steps. The kinetic properties and thermal stability of purified LDH were analyzed, revealing significant differences between the two enzyme forms in Vmax, Km pyruvate, and I50 pyruvate as well as melting temperature determined by differential scanning fluorimetry. The phosphorylation state of aerobic and anoxic forms of LDH was visualized by ProQ Diamond phosphoprotein staining, the results indicating that the anoxic form had a higher phosphorylation state. Incubation studies that promoted protein kinase versus protein phosphatase actions showed that changes in the phosphorylation state of aerobic and anoxic forms mimicked the anoxia-responsive changes in Km pyruvate and I50 pyruvate. The high phosphate form of liver LDH that occurs in anoxic turtles appears to be a less active form. Turtle liver LDH was also subject to another form of posttranslational modification, protein acetylation, with a 70% higher content of acetylated lysine residues on anoxic versus aerobic LDH. This is the first study to show that LDH function in an anoxia-tolerant animal can be differentially modified between aerobic and anoxic states via the mechanism of posttranslational modification. [Copyright &y& Elsevier]

Abstract: Burkholderia pseudomallei infections are fastidious to treat with conventional antibiotic therapy, often involving a combination of drugs and long-term regimes. Bacterial genetic determinants contribute to the resistance of B. pseudomallei to many classes of antibiotics. In addition, anaerobiosis and hypoxia in abscesses typical of melioidosis select for persistent populations of B. pseudomallei refractory to a broad spectrum of antibacterials. We tested the susceptibility of B. pseudomallei to the drugs hydroxyurea, spermine NONOate and DETA NONOate that release nitric oxide (NO). Our investigations indicate that B. pseudomallei are killed by NO in a concentration and time-dependent fashion. The cytoxicity of this diatomic radical against B. pseudomallei depends on both the culture medium and growth phase of the bacteria. Rapidly growing, but not stationary phase, B. pseudomallei are readily killed upon exposure to the NO donor spermine NONOate. NO also has excellent antimicrobial activity against anaerobic B. pseudomallei. In addition, persistent bacteria highly resistant to most conventional antibiotics are remarkably susceptible to NO. Sublethal concentrations of NO inhibited the enzymatic activity of [4Fe–4S]-cofactored aconitase of aerobic and anaerobic B. pseudomallei. The strong anti-B. pseudomallei activity of NO described herein merits further studies on the application of NO-based antibiotics for the treatment of melioidosis. [Copyright &y& Elsevier]

Benthic marine organisms such as mollusks are often exposed to periodic oxygen deficiency (due to the tidal exposure and/or seasonal expansion of the oxygen-deficient dead zones) and pollution by metals [e.g., cadmium, (Cd)]. These stressors can strongly affect mollusks' survival; however, physiological mechanisms of their combined effects are not fully understood. We studied the effects of Cd exposure on metabolic responses to prolonged anoxia and subsequent recovery in anoxia-tolerant intertidal mollusks Crassostrea virginica (eastern oysters). Anoxia led to an onset of anaerobiosis indicated by accumulation of L-alanine, acetate, and succinate. Prolonged anoxia (for 6 days) caused a decline in the maximum activity of electron transport chain and ADP-stimulated (state 3) oxygen uptake by mitochondria (MO2), but no change in the resting (state 4) MO2 of oyster mitochondria, along with a slight but significant reduction of mitochondrial respiratory control ratio. During reoxygenation, there was a significant overshoot of mitochondrial MO2 (by up to 70% above the normoxic steady-state values) in control oysters. Mild mitochondrial uncoupling during prolonged shutdown in anoxic tissues and a subsequent strong stimulation of mitochondrial flux during recovery may help to rapidly restore redox status and protect against elevated reactive oxygen species formation in oysters. Exposure to Cd inhibits anaerobic metabolism, abolishes reoxygenation-induced stimulation of mitochondrial MO2, and leads to oxidative stress (indicated by accumulation of DNA lesions) and a loss of mitochondrial capacity during postanoxic recovery. This may result in increased sensitivity to intermittent hypoxia and anoxia in Cd-exposed mollusks and will have implications for their survival in polluted estuaries and coastal zones. [ABSTRACT FROM AUTHOR]

Ducks are well-known to be more tolerant to asphyxia than non-diving birds, but it is not known if their defences include enhanced neuronal hypoxia tolerance. To test this, we compared extracellular recordings of spontaneous activity in the Purkinje cell layer of 400 μm thick isolated cerebellar slices from eider ducks, chickens and rats, before, during and after 60 min hypoxia (95%N2–5%CO2) or chemical anoxia (hypoxia + 2 mM NaCN). Most slices rapidly lost activity in hypoxia, with or without recovery after rinse and return to normoxia (95%O2–5%CO2), but some maintained spontaneous activity throughout the insult. Proportions of ‘surviving’ (i.e. recovering or active) duck slices were significantly higher than for chickens in anoxia, and relative activity levels were higher for ducks than for chickens during hypoxia, anoxia and recovery. Survival of rat slices was significantly poorer than for birds under all conditions. Results suggest that (1) duck cerebellar neurons are intrinsically more hypoxia-tolerant than chicken neurons; (2) avian neurons are more hypoxia-tolerant than rat neurons, and (3) the enhanced hypoxic tolerance of duck neurons largely depended on efficient anaerobiosis since it mainly manifested itself in chemical anoxia. Mechanisms underlying the observed differences in neuronal hypoxic responses remain to be elucidated. [ABSTRACT FROM AUTHOR]

For decades altitude training has been used by endurance athletes and coaches to enhance sea-level performance. Whether altitude training does, in fact, enhance sea level performance and, if so, by what means has been the subject of a number of investigations. Data produced principally by Levine and Stray-Gundersen have shown that living for 4 weeks at 2500 m, while performing the more intense training sessions near sea level will provide an average improvement in sea level endurance performance (duration of competition: 7–20 min) of approximately 1.5%, ranging from no improvement to 6% improvement. This benefit lasts for at least 3 weeks on return to sea level. Two mechanisms have been shown to be associated with improvement in performance. One is an increase in red cell mass (∼8%) that results in an improved maximal oxygen uptake (∼5%). That must be combined with maintenance of training velocities and oxygen flux to realize the improvement in subsequent sea level performance. We find no evidence of changes in running economy or markers of anaerobic energy utilization. Our results have been obtained in runners ranging from collegiate to elite. Wehrlin et al. have recently confirmed these results in elite orienteers. While there are no specific studies addressing the use of living high, training low in football players, it is likely that an improvement in maximal oxygen uptake, all other factors equal, would enhance football performance. This benefit must be weighed against the time away (4 weeks) from home and competition necessary to gain these benefits. [ABSTRACT FROM AUTHOR]

With the use of technologically constructed hypoxic environments (TCHE) in soccer as our case, we propose four check points from which to evaluate new performance-enhancing technologies in sport. These are (I) Is the technology beneficial, (II) Is it safe, (III) Can fairness be assured, and (IV) Is the technology in line with the spirit of or rationale for sport? The use of TCHE is ambiguous. On the one hand, in situations with grave inequalities between teams due to lack of acclimatization of one team, TCHE can be an efficient means to even the playing field and out of concern for athlete welfare and health. On the other hand, if used as a pure performance-enhancing means to enhance the oxygen-carrying capacity of the blood independent of altitude, it belongs to a category of expert-assisted performance enhancement that may challenge athletic autonomy and the responsibility for one's own performance and, hence, the spirit of sport. [ABSTRACT FROM AUTHOR]

Freezing survival was assessed in three species of terrestrial slugs, a holarctic but native North American species, Deroceras laeve, and two species introduced from Europe, D. reticulatum and Arion circumscriptus. The introduced species showed very poor freezing survival. Supercooling points of the introduced species were quite high (∼−3°C) and their freezing survival was very poor, limited to short-term freezing at −1.2 to −1.5°C and low ice contents (23–44%). D. laeve showed a significant elevation of supercooling point between slugs collected in the autumn (−4.8°C ± 0.5) and those collected early in the spring (−3.1°C ± 0.4). This species also showed substantial freezing survival which was greater for spring-collected slugs (100% survival of 1 h freezing at −2°C with an ice content of 65%) than for autumn animals (100% survival for 1 h at −1°C with ∼40% ice). Carbohydrate and amino acid responses to freezing and anoxia exposures were compared in the two Deroceras species. D. laeve showed a strong hyperglycemic response to freezing, a 100-fold increase in glucose levels that suggested that glucose may have a cryoprotective function in this species. D. reticulatum did not accumulate glucose and neither species produced glycerol or lactate. Both species showed typical responses to anoxia (aspartate and glutamate catabolism, alanine and succinate accumulation) and D. laeve also showed this pattern during freezing, suggesting a natural switch to anaerobiosis to support freezing survival. [ABSTRACT FROM AUTHOR]

The purpose of this study was to examine the responses of the oyster Crassostrea gigas to oxygen levels at subcellular and whole organism levels. Two experiments were carried out. The first experiment was designed to measure the clearance and oxygen consumption rates of oysters exposed at different concentrations of oxygen at 15, 20 and 25°C for 20 h. The goal of this first part was to estimate the hypoxic threshold for oysters below which their metabolism shifts towards anaerobiosis, by estimating the oxygen critical point (PcO2) at 15, 20 and 25°C. The second experiment was carried out to evaluate the metabolic adaptations to hypoxia for 20 days at three temperatures: 12, 15 and 20°C. The metabolic pathways were characterized by the measurement of the enzymes pyruvate kinase (PK) and phosphoenolpyruvate carboxykinase (PEPCK), the alanine and succinate content and the adenylate energy charge. Respiratory chain functioning was estimated by the measurement of the activity of the electron transport system (ETS). The values of PcO2 were 3.02±0.15, 3.43±0.20 and 3.28±0.24 mg O2 l-1 at 15, 20 and 25°C, respectively. In whole oysters, hypoxia involved the inhibition of PK whatever the temperature, but PEPCK was not stimulated. Succinate accumulated significantly only at 12°C and alanine at 12 and 15°C. A negative relationship between the PK activity and the alanine content was only found in hypoxic oysters. Finally, hypoxia increased significantly the activity of ETS. With high PcO2 values, the metabolic depression occurred quickly, showing that oysters had a low capacity to regulate their respiration when oxygen availability is reduced, particularly in the summer. [ABSTRACT FROM AUTHOR]

The annual killifish Ausirofundulus limnaeus survives in ephemeral pond habitats by producing drought-tolerant diapausing embryos. These embryos probably experience oxygen deprivation as part of their normal developmental environment. We assessed the anoxia tolerance of A. limnaeus embryos across the duration of embryonic development. Embryos develop a substantial tolerance to anoxia during early development, which peaks during diapause II. This extreme tolerance of anoxia is retained during the first 4 days of post-diapause II development and is then lost. Metabolism during anoxia appears to be supported mainly by production of lactate, with alanine and succinate production contributing to a lesser degree. Anoxic embryos also accumulate large quantities of γ-arninobutyrate (GABA), a potential protector of neural function. It appears that the suite of characters associated with normal development and entry into diapause II in this species prepares the embryos for long-term survival in anoxia even while the embryos are exposed to aerobic conditions. This is the first report of such extreme anoxia tolerance in a vertebrate embryo, and introduces a new model for the study of anoxia tolerance in vertebrates. [ABSTRACT FROM AUTHOR]

Summary: The relationship between coleoptile elongation and ethanolic fermentation was investigated in rice (Oryza sativa L.) coleoptiles of four cultivars subjected to a 48-h anoxic stress. The coleoptile elongation of all cultivars was suppressed by anoxic stress; however, the elongation of cvs Yukihikari and Nipponbare was much greater than that of cvs Leulikelash and Asahimochi. The stress did not significantly increase lactate dehydrogenase (LDH) activity or lactate concentration, but increased alcohol dehydrogenase (ADH) and pyruvate decarboxylase (PDC) activities, as well as ethanol concentration in the coleoptiles of all cultivars. The elevated ADH and PDC activities and ethanol concentration in cvs Yukihikari and Nipponbare were much greater than those of cvs Leulikelash and Asahimochi, suggesting that ethanolic fermentation is likely more active in cvs Yukihikari and Nipponbare than in cvs Leulikelash and Asahimochi. ATP concentration in cvs Yukihikari and Nipponbare in anoxia was also greater than that in cvs Leulikelash and Asahimochi in anoxia. The ethanol concentration in the coleoptiles was correlated with anoxia tolerance with respect to the ATP concentration and coleoptile elongation. These results suggest that the ability to increase ethanolic fermentation may be one of the determinants in anoxia tolerance of rice coleoptiles. [Copyright &y& Elsevier]

Hemoglobin gene expression is upregulated during hypoxia. To determine whether the induction occurs via similar mechanisms that have been proposed for other hypoxically induced proteins, barley ( Hordeum vulgare L.) aleurone layers were treated with various agents that interfere with known components of signal transduction. Ruthenium red, an organelle calcium channel blocker, inhibited anoxia-induced hemoglobin (Hb) and alcohol dehydrogenase (EC 1.1.1.1) (Adh) gene expression in a dose-dependent manner. The divalent ionophore, A23187, combined with EGTA also dramatically reduced anoxia-induced Hb and Adh expression. Normal induction of Hb by anoxia in EGTA-treated cells was restored by adding exogenous Ca2+ but not Mg2+, suggesting that cytosolic calcium is involved in Hb and Adh regulation. W-7, a calmodulin antagonist, did not affect anaerobically induced Hb and Adh expression even though it induced Hb under aerobiosis. A3, a protein kinase inhibitor, did not significantly affect anaerobically induced Hb, but did significantly upregulate the gene under aerobic conditions. The results indicate that calmodulin-independent anaerobic alteration in cytosolic Ca2+ and protein dephosphorylation are factors in Hb induction. [ABSTRACT FROM AUTHOR]

Abstract: The responses of members of the three main MAPK families (ERK, JNK/SAPK, p38 MAPK), as well as selected peripheral pathways, were examined in hepatopancreas of the marine periwinkle, Littorina littorea, to determine if anoxia exposure influenced the total protein content or the phosphorylation status of any key components. The content of active phospho-p38 MAPK was 2-fold higher in hepatopancreas from anoxic snails relative to controls. A 1.7-fold increase in the amount of phospho-Hsp27 and a 1.3-fold increase in phospho-CREB correlated well with the changes in p38 MAPK phosphorylation. Activation of these factors via p38 MAPK may be vital to the reorganization of metabolic responses to anoxia in hepatopancreas. No changes in components of the JNK/SAPK and ERK pathways occurred and transcription factors involved in lipid metabolism did not appear to be affected by anoxia. The present analysis of a variety of signaling pathways has implicated the p38 MAPK pathway as a key anoxia-responsive signal transduction pathway in L. littorea. [Copyright &y& Elsevier]

Abstract: Short-term anoxic treatments have been used as an inexpensive non-chemical technology to prevent physiological disorders and delay ripening in fruits. However, treatment efficacy was shown to depend on the intensity of the application and on the type of commodity treated. The present research was carried out to investigate the effect of anoxic treatments on the ethanol and acetaldehyde content in Summerset peaches. Fruit were placed under N2 atmosphere for different combinations of time and temperature. Ethanol and acetaldehyde concentrations were measured after treatment application, and periodically during storage under aerobic conditions. Ethanol production was analyzed by response surface methodology, with time and temperature of exposure to anoxia being the independent variables. Effect of anoxia on flesh firmness was positive, since all treatments slowed fruit softening. The combination of 64h at 20°C induced the highest ethanol and acetaldehyde concentration after treatment, and throughout storage, though values sharply decreased when fruit were held at 20°C following storage. Ethanol was a good variable for monitoring the intensity of an anoxic treatment, with its production closely linked to the severity of the stress exerted by the various treatments on the fruit. The fitted quadratic response surface had a high R-square value, with 96.6% of the ethanol accumulation being explained by the model. [Copyright &y& Elsevier]

This review attempted to follow the establishment of a novel branch of biology arisen at the interfaces between plant physiology, biochemistry, and molecular biology—plant anaerobic stress. Most attention was given to the early period of these investigations, the activity of the members of International Society for Plant Anaerobiosis in particular, and the contribution of Russian scientists, who played a significant role at that time in the establishment and international recognition of this new trend. In this connection, the following points are considered: (1) Crawford's metabolic theory, which could not withstand experimental verification but induced an active discussion, thus stimulating further investigations in this field; (2) a concept of two main strategies of plant adaptation to anaerobic stress (true and apparent adaptation), which was put forward based on the following experimental data: (a) a discovery of a paradoxical phenomenon of hyper-sensitivity, but not hyper-resistance to anoxia, of the flood-tolerant plant roots (“apparent” tolerance); (b) the elucidation of the physiological role of oxygen transported from aerated organs of flood-tolerant plants to the roots inhabiting anaerobic environment; (c) demonstration of the key role of both energy metabolism, and (d) substrate providing for glycolysis and ethanolic fermentation in plants manifesting “true” tolerance to oxygen deprivation; (3) the discovery of plant stress proteins; and finally (4) pH-stat theory put forward by Davies. [ABSTRACT FROM AUTHOR]

The tiger beetle, Phaeoxantha klugii Chaudoir, survives the annual inundation period in Central Amazonian floodplains as a third-instar larvae submerged in the soil at approximately 29°C for up to 3.5 months. Previous studies showed an exceptional anoxia resistance in these larvae and this study investigates whether they perform anaerobiosis. Larvae collected in the field were exposed to a pure nitrogen atmosphere for 0-9 days in the laboratory. The content of lactate, alanine, free sugars and glycogen is analysed in surviving larvae. Lactate and alanine contents rise during anoxia from around 1.5 and 7 to 6-14 and 15-22 µmol g-1 fresh mass, respectively, providing evidence for anaerobic metabolism. Both compounds show a steep increase during the first 12 h and a tendency to rise further with increasing duration of anoxic conditions, indicating a significant metabolic depression within the first day. Content of free sugars and glycogen varies greatly between individuals and ranges from 0.08-2.5 and 0.05-2.9 mg g-1 fresh mass, respectively. Whether glycogen is used as metabolic substrate for anaerobiosis could not be verified. The findings for free sugars indicate that larvae apparently maintain the ability to regulate the level of glucose and/or trehalose even after 9 days of anoxia. [ABSTRACT FROM AUTHOR]

Peter W. Hochachka led a grand life of science adventure and left as his legacy a whole new field—biochemical adaptation. Oxygen was at the core of Peter''s career and his laboratory made major contributions to our understanding of how animals deal with variation in oxygen availability in many forms. He analyzed the molecular mechanisms that support facultative anaerobiosis, studied muscle exercise metabolism for high speed flight, swimming and running, investigated mammalian diving on many trips to the Antarctic to study Weddell seals, and probed the metabolic and genetic adaptations that provide optimal hypoxia tolerance for humans residing at high altitudes. His work illuminated both biochemical and physiological mechanisms that are used to optimize aerobic metabolism, to compensate for hypoxic insults, and to conserve energy by strong metabolic rate depression under anoxia. His articles, books and lectures galvanized the field with leading-edge insights and theories and he consistently challenged comparative biochemists to use their unique model systems to explore the range and breadth of animal strategies of biochemical adaptation. Lessons drawn from my training in Peter''s laboratory have led me on continuing explorations of adaptations in enzyme function, signal transduction, gene expression, and antioxidant defenses ranging over systems of anoxia tolerance, freezing survival, estivation, and mammalian hibernation. [Copyright &y& Elsevier]

The activity of alcohol dehydrogenase (ADH) was measured in corms and roots of the submerged freshwater macrophyte Isoetes alpinus Kirk. growing in situ, and related to its capacity for internal oxygen transport and to carbohydrate translocation. ADH activity was present in roots but not corms at uniform activity (0.15–0.35 × 10−6 mol g−1 fresh weight s−1) over the entire plant depth range (3–7 m depth), and was intermediate to that developed in excised roots after 1-week exposure to either dissolved oxygen at air-saturation or to anoxia. Responses of photosynthesis and root oxygen release to light intensity confirmed that shoot-to-root oxygen transport saturated at similar light intensities to photosynthetic oxygen evolution, but was positive in the dark and at irradiances below the compensation point for photosynthesis, due to contributions to transport by oxygen diffusion from the external medium. Transport of 14C-labelled photo-assimilates to roots nevertheless ceased when intact plants were exposed to a combination of leaf darkness and root external anoxia, even when high 14C concentrations were present in shoots, but remained high when the roots were provided with external oxygen. The lack of any control over permeability of the root surface to gases in this species suggested that ADH activity and reduced translocation is most likely caused by development of hypoxic tissues in the apical tissue. These results suggest that reductions in ambient light intensity may have indirect effects on I. alpinus viability by increasing the degree of root hypoxia and impairing carbon partitioning. [ABSTRACT FROM AUTHOR]

An important functional advantage of hypoxic hypometabolism is that it blunts the acid–base consequences of hypoxia. Hypoxia can lead to anaerobiosis and metabolic acidosis and, in animals that are apneic, to respiratory acidosis. A fall in blood and tissue pH is a major limiting factor in hypoxic tolerance and a variety of strategies occur in vertebrates, in concert with hypometabolism, to respond to this acid–base challenge. These include sequestering of lactic acid away from the circulating blood during the hypoxic exposure, either in underperfused tissues or in mineralized tissues, supplementing extracellular buffering by releasing bone mineral into the circulation, and utilizing alternative metabolic pathways for anaerobiosis to produce ethanol rather than lactate as the principal end-product. For submerged air-breathing ectotherms, effective cutaneous O2 and CO2 exchange can also allow an animal to avoid or minimize both anaerobiosis and respiratory acidosis. These responses serve to maintain a viable acid–base state in the body and to extend the time that the hypoxic stress can be endured. [Copyright &y& Elsevier]

Terrestrial invertebrates experience hypoxia in many habitats and under a variety of physiological conditions. Some groups (at least insects) are much more capable of recovery from anoxia than most vertebrates, but there is still a tremendous unexplained variation in hypoxia/anoxia tolerance among terrestrial invertebrates. Crustaceans and arachnids may be less often confronted with hypoxic environments than insects and myriapods and also seem to be less hypoxia/anoxia tolerant. Tracheated groups, especially those that are able to ventilate their tracheal system like many insects, cope with lower critical PO2 than nontracheated groups. Modulation of oxygen carrier proteins is normally not important in hypoxia resistance. Recent application of genetic and cellular tools are revealing that many of the same pathways documented for mammals (e.g. HIF, nitric oxide) function to regulate morphological and biochemical responses to hypoxia/anoxia in insects. [Copyright &y& Elsevier]

Tolerance to low oxygen availability is likely to be due to the interaction of several factors. Sugar availability is one of the elements required to support anaerobic metabolism. In cereal grains the availability of soluble sugars is limited, while starch is stored in large amounts. Degradation of starch under anoxia is therefore needed to avoid sugar starvation leading to rapid cell death. The striking difference in the ability to produce α‐amylase when comparing the anoxia‐tolerant rice (Oryza sativa L.) grains with grains of other cereals is not easily explained. Rice is able to respond to gibberellins under anoxia, but the response is too slow to explain the rapid production of α‐amylase enzyme. In the present work we demonstrated that α‐amylase production during the first 2 d after imbibition is mostly due to the activity of the Ramy3D gene, encoding for the G and H isoforms of α‐amylase. The induction of Ramy3D transcription is likely to result from a low sugar content in the grains incubated under anoxia. The ability of rice embryos to sense sugars under anoxia is reported. [ABSTRACT FROM PUBLISHER]

Sterol esterification in Saccharomyces cerevisiae is catalyzed by two acyl-CoA:sterol acyltransferases encoded by the genes ARE1 and ARE2. Using double mutants in the HEM1 gene and individual ARE genes we demonstrated that the relative contribution of these two enzymes to sterol esterification was dependent on cellular heme status. Observed changes in sterol esterification could be explained by a different effect of heme on the transcription of both genes: while the ARE1 transcript level was elevated in heme-deficient and anaerobic cells, the ARE2 gene transcript was more abundant in aerobic cells competent for heme synthesis. Our results indicate that transcriptional regulation of ARE genes by heme and specific substrate preferences of Are1p and Are2p may be involved in the adaptation of yeast sterol metabolism to hypoxia. [Copyright &y& Elsevier]

In this study, we compared survivorship, heat dissipation and biochemical features of anaerobiosis of two tiger beetle species (Coleoptera: Cicindelidae) exposed to anoxia. One species commonly experiences environmental immersion from rainfall and snowmelt (Cicindela togata), and the habitat of the other (Amblycheila cylindriformis) is not prone to flooding. The ancestral genus, A. cylindriformis, survives anoxia for only 2 days at 25 °C. In response to anoxia, these larvae immediately lose locomotory abilities, tissue concentrations of ATP fall precipitously within 12 h, and significant amounts of lactate are quickly produced. In contrast, C. togata larvae tolerate anoxia for 5 days. Heat dissipation is downregulated to a greater degree than that seen in A. cylindriformis (3.4% versus 14% of standard normoxic rate, respectively), the ability for locomotion is maintained and normoxic levels of ATP are defended for at least 24 h. Lactate is not accumulated until well into anoxic bout, and significant amounts of alanine are also produced. This study provides evidence that tiger beetles differ in physiological responses to anoxia, and that these differences are correlated with flooding risk and with species distribution. [ABSTRACT FROM AUTHOR]

The responses of two aquatic plants, arrowhead (Sagittaria pygmaea Miq.) and pondweed (Potamogeton distinctus A. Benn), to anoxia were compared with those of rice (Oryza sativa L.). Shoot elongation of arrowhead tubers was enhanced at around 1 kPa O2, whereas that of pondweed turions was slight in air and reached a maximum in the absence of O2. Anaerobic enhancement of alcohol dehydrogenase (ADH) activity took place in rice coleoptiles but not in arrowhead and pondweed shoots. Shoots of both arrowhead and pondweed maintained a more stable energy status than did the rice coleoptile under anaerobic conditions. Total adenylate nucleotide contents of arrowhead and pondweed shoots were constant under anaerobic conditions. Adenylate energy charge in both shoots remained at a high and stable level of more than 0·8 for at least 8 d. Three forms of ADH from arrowhead shoots were separated by starch gel electrophoresis, showing that the activity of each ADH form was different under aerobic and anaerobic conditions. The incorporation of 35S-labelled Cys and Met into soluble proteins in arrowhead shoots showed active protein biosynthesis and an involvement of a special set of polypeptides in the anaerobiosis. [ABSTRACT FROM AUTHOR]

The effects of contingent feedback (CF), non-contingent feedback (NCF), and no feedback (NFl on heart rate (HR) variability control were studied. Nine matched trios of subjects were given training in HR variability control on 3 consecutive days. CF and NCF subjects attempted HR variability control while HR feedback was continuously presented, and NF subjects attempted HR variability control without feedback. During the training periods the CF group produced reliably lower mean HR variance than the NF group, but the NCF group was not reliably different from either of those groups. There was no significant difference among groups in negative linear trend across training periods, and the CF group did not continue to produce decreasing HR variances over sessions. However, the CF and NCF groups showed reliable quadratic trends, Indicating an initial drop in HR variance followed by a return to pretraining levels. Both CF and NCF groups showed rapid, reliable increases in respiration rate during training periods. The results pose serious problems for studies which have reported feedback-related control of HR variability. [ABSTRACT FROM AUTHOR]

Alcohol dehydrogenase (ADH; EC 1.1.1.1) activity was measured in Picea glauca (Moench) Voss cell suspensions under differing conditions of hypoxia. ADH activity increased 4.5 fold after 48 h of induction. When cells were induced under different levels of hypoxia (2,5 and 20% O2) changes in ADH activity were found to increase with lower levels of oxygen. Alanine aminotransferase (AlaAT; EC 2.6.1.2) activity increased under hypoxia in a pattern similar to ADH, however lactate dehydrogenase (LDH; EC 1.1.1.27) activity did not increase under hypoxic conditions. The ability of white spruce cells to accurately regulate heterologous anaerobic promotors was tested by electroporating chimeric ADH reporter genes into protoplasts. While protoplasts were capable of anaerobically regulating a maize ADH reporter construct, constructs with dicotyledonous promoters (pea and Arabidopsis) were not expressed. [ABSTRACT FROM AUTHOR]

To study the transcriptional and translational responses that occur in rice (Oryza sativa L., cv. Arborio) coleoptiles and roots during the first hours of anoxia, we analysed the patterns of in vitro translational products and in vivo labelled newly synthesized proteins obtained from both organs grown in the presence or absence of O2. Analysis of the protein pattern obtained from nuclear-enriched subcellular fractions showed the enhancement of the synthesis of several proteins with molecular weights of 67-62,54 and 34 kD. In the cytoplasmic-enriched subcellular fractions, we found that the synthesis of polypeptides of relatively low molecular weight (13-16 kD) was strongly induced by anaerobiosis. In vitro translation analysis showed that this enhancement was mainly due to a rapid increase in the level of the corresponding mRNA species. These low- molecular-weight translational products were already detected after 1 h of anoxia in both rice roots and rice coleoptiles, but remained undetectable in aerobically grown organs. Additional anaerobic-specific transcrip- tional and translational changes were also observed. In particular, a specific reduction in the level of mRNAs coding for polypeptides of 38 kD occurred in both roots and coleoptiles after only 1 h of anoxia. [ABSTRACT FROM AUTHOR]

The temperate seagrass Zostera marina L. typically grows in highly reducing sediments. Photosynthesis-mediated O supplied to below-ground tissues sustains aerobic respiration during photosynthetic periods. Roots, however, experience daily periods of anoxia and/or hypoxia at night and under conditions that reduce photosynthesis. Rhizosphere cores of Z. marina were collected in August 1984 from Great Harbor, Massachusetts, USA. We examined short-term anaerobic metabolism of [U-C]sucrose in excised roots and roots of intact plants. Under anaerobic conditions roots showed appreciable labeling of CO, ethanol and lactate, and slight labeling of alanine and other metabolites. Over 95% of the C-ethanol was recovered in the root exudate. Release of other metabolites from the roots was minimal. Ethanol was also released from hypoxic/anoxic roots of intact plants and none of this ethanol was transported to the shoot under any experimental conditions. Loss of ethanol from roots prevented tissue levels of this phytotoxin from increasing during anaerobiosis despite increased synthesis of ethanol. Anaerobic metabolism of [U-C]glutamate in excised roots led to appreciable labelling of γ-aminobutyrate, which was known to accumulate in eelgrass roots. Roots recovered to fully aerobic metabolism within 4 h after re-establishment of aerobic conditions. The contributions of these root metabolic responses to the ability of Z. marina to grow in reducing marine sediments are related to light-regulated interactions of shoots and roots that likely dictate depth penetration, distribution and ecological success of eelgrass. [ABSTRACT FROM AUTHOR]

In Ensis directus (Conrad) metabolic changes accompanying muscular activity and experimental anoxia were investigated. Octopine dehydrogenase, strombine dehydrogenase, alanopine dehydrogenase and lactic dehydrogenase were shown to be present in different parts of the body, but the activity of ODH always exceeded that of the other enzymes by far. Muscular activity connected with 'escape digging' or evoked by electrical stimulation resulted in a decrease of stored phosphagen and in an accumulation of octopine. During recovery the normal concentrations of energy-rich phosphates were restored quickly, whereas octopine was removed more slowly. In experimental anoxia only minor quantities of L-and D-alanine, succinate, propionate, and acetate were formed. Quantitatively the most important endproduct was strombine, which rose linearly with the duration of anoxic incubation. [ABSTRACT FROM AUTHOR]

Different plant species vary widely in their degree of tolerance to anaerobic stress. The precise molecular mechanisms underlying such tolerance are unknown, but both enzymes of the alcoholic fermentation pathway, pyruvate decarboxylase and alcohol dehydrogenase, are strongly induced during conditions of low oxygen. We have isolated two adh and two pdc genes in Arabidopsis thaliana (L.)Heynh. The patterns of expression of these genes have identified two sets of alcoholic fermentation pathway genes, each of which might play a different role in the adaptive response of the plant to anoxia. One set is strongly induced by low oxygen stress mainly in the roots, while the other set is expressed constitutively in both roots and leaves. Arabidopsis also has two haemoglobin genes, only one of which is induced by hypoxia. We are investigating the components of the signal transduction pathways leading to adh and pdc induction. Implications of our findings for metabolic engineering of plants to improve flooding tolerance are discussed. [ABSTRACT FROM AUTHOR]

Plants are aerobic organisms that require oxygen for their respiration. Hypoxia arises due to the insufficient availability of oxygen, and is sensed by plants, which adapt their growth and metabolism accordingly. Plant hypoxia can occur as a result of excessive rain and soil waterlogging, thus constraining plant growth. Increasing research on hypoxia has led to the discovery of the mechanisms that enable rice to be productive even when partly submerged. The identification of Ethylene Response Factors (ERFs) as the transcription factors that enable rice to survive submergence has paved the way to the discovery of oxygen sensing in plants. This, in turn has extended the study of hypoxia to plant development and plant–microbe interaction. In this review, we highlight the many facets of plant hypoxia, encompassing stress physiology, developmental biology and plant pathology. [ABSTRACT FROM AUTHOR]

Overexpression of phytoglobins (formerly plant hemoglobins) increases the survival rate of plant tissues under hypoxia stress by the following two known mechanisms: (1) scavenging of nitric oxide (NO) in the phytoglobin/NO cycle and (2) mimicking ethylene priming to hypoxia when NO scavenging activates transcription factors that are regulated by levels of NO and O2 in the N-end rule pathway. To map the cellular and metabolic effects of hypoxia in barley (Hordeum vulgare L., cv. Golden Promise), with or without priming to hypoxia, we studied the proteome and metabolome of wild type (WT) and hemoglobin overexpressing (HO) plants in normoxia and after 24 h hypoxia (WT24, HO24). The WT plants were more susceptible to hypoxia than HO plants. The chlorophyll a + b content was lowered by 50% and biomass by 30% in WT24 compared to WT, while HO plants were unaffected. We observed an increase in ROS production during hypoxia treatment in WT seedlings that was not observed in HO seedlings. We identified and quantified 9694 proteins out of which 1107 changed significantly in abundance. Many proteins, such as ion transporters, Ca2+-signal transduction, and proteins related to protein degradation were downregulated in HO plants during hypoxia, but not in WT plants. Changes in the levels of histones indicates that chromatin restructuring plays a role in the priming of hypoxia. We also identified and quantified 1470 metabolites, of which the abundance of >500 changed significantly. In summary the data confirm known mechanisms of hypoxia priming by ethylene priming and N-end rule activation; however, the data also indicate the existence of other mechanisms for hypoxia priming in plants. [ABSTRACT FROM AUTHOR]

The imposition of anaerobic conditions resulted in a rapid (2 min) and transient increase in cGMP content in root and coleoptile of rice seedlings. The level of cAMP was strongly decreased by anoxia. These data are discussed in relation to the anaerobic response. [ABSTRACT FROM AUTHOR]