EPR evidence for generation of hydroxyl radical triggered byN-acetylchitooligosaccharide elicitor and a protein phosphatase inhibitor in suspension-cultured rice cells

Summary N-acetylchitooligosaccharides, fragments of the backbone of fungal cell wall, trigger rapid membrane responses such as transient depolarization, and elicit defense reactions including phytoalexin production in suspension-cultured rice cells. The generation of reactive oxygen species triggered by the oligosaccharide signal was analyzed with EPR spectroscopy using a spin trapping system, 4-pyridyl 1-oxideN-tert-butyl nitrone (4-POBN) and ethanol. OH generation was detected as the -hydroxyethyl adduct of 4-POBN after elicitation. Superoxide dismutase, catalase or diethylenetriamine pentaacetic acid, a metal chelator, inhibited generation, proposing the following reaction sequence: generation of in response to the oligosaccharide elicitor, followed by dismutation to H₂O₂, then generation of by the reaction of H₂O₂ with Fe²⁺ that is generated by the reduction of Fe³⁺ by . Generation of the same reactive oxygen species was also triggered by calyculin A, a protein phosphatase inhibitor, alone, suggesting the involvement of protein phosphorylation in its regulation during the oligosaccharide signal transduction.Abbreviations DMPO 5,5-dimethyl-1-pyrroline N-oxide - DTPA diethylenetriamine pentaacetic acid - 4-POBN 4-pyridyl 1-oxideN-tert-butylnitrone - SOD Superoxide dismutase - 4-hydroxy-TEMPO 2,2,6,6-tetramethyl-4-hydroxypiperidine-1-oxyl     

Ionol (BHT) produces superoxide anion

In aqueous medium etiolated wheat seedlings release superoxide anion ( ). Interaction of a synthetic antioxidant, butylated hydroxytoluene (BHT, ionol), with oxygen in the aqueous medium is accompanied by formation. This suggests that under certain conditions BHT behaves as a prooxidant. A natural antioxidant, superoxide dismutase (SOD), and also a wound healing preparation, emulsified denatured placenta (EDP), do not exhibit the prooxidant properties. In contrast to BHT, they reduce production by the etiolated wheat seedling system.     

Contribution of a plasma membrane redox system to the superoxide production by wheat root cells

Summary Wound stress activated wheat root cells to produce oxygen radicals. The production was accompanied by an increased permeability for potassium ions and a depolarization of the plasma membrane. Various electron donors, such as the nonpenetrating donor potassium ferrocyanide as well as NADH and NADPH, caused the amplification of superoxide production by root cells. The -generating system in wheat root cells was found to be considerably sensitive to diphenylene iodonium, which is generally considered as a suicide inhibitor of neutrophil NADPH oxidase, and to other inhibitors of flavoprotein activity, chlorpromazine and quinine. The xenobiotic compound amidopyrine caused activation of the -generating system, which was depressed by DPI. The -generating system in root cells was shown to be significantly dependent on calcium content. Calcium loading of the root cells induced a powerful increase of the superoxide release. Data obtained indicate that superoxide generation is one of the early events of the wound stress response. Redox systems of the plasma membrane may be involved in the superoxide production in response to wound stress and detoxification of xenobiotic compounds in root cells.Abbreviations DPI diphenylene iodonium - MP membrane potential - superoxide anion radical - ROS reactive-oxygen species - SOD superoxide dismutase     

Effect of electron-transport inhibitors on the generation of reactive oxygen species by pea mitochondria during succinate oxidation

The effect of inhibitors of the cytochrome pathway and alternative oxidase on the rate of respiration and generation of reactive oxygen species by pea mitochondria was studied. Respiration of mitochondria from pea cotyledons was inhibited by 70-80% by salicylhydroxamate (SHAM). The rate of hydrogen peroxide production by pea cotyledon mitochondria during succinate oxidation was 0.15 nmol/min per mg protein. SHAM considerably accelerated the hydrogen peroxide production. The SHAM-dependent H₂O₂ production was stimulated by 2 M antimycin A and inhibited by 5 mM KCN and 1 M myxothiazol. The study of the rate of generation by pea mitochondria using EPR spin traps and epinephrine oxidation showed that H₂O₂ accumulation can be accounted for by a significant increase in the rate of production.     

Generation of superoxide-radical by the NADH: Ubiquinone oxidoreductase of heart mitochondria

Besides major NADH-, succinate-, and other substrate oxidase reactions resulting in four-electron reduction of oxygen to water, the mitochondrial respiratory chain catalyzes one-electron reduction of oxygen to superoxide radical followed by formation of hydrogen peroxide. In this paper the superoxide generation by Complex I in tightly coupled bovine heart submitochondrial particles is quantitatively characterized.The rate of superoxide formation during -controlled respiration with succinate depends linearly on oxygen concentration and contributes approximately 0.4% of the overall oxidase activity at saturating (0.25 mM) oxygen. The major part of one-electron oxygen reduction during succinate oxidation (80%) proceeds via Complex I at the expense of its -dependent reduction (reverse electron transfer). At saturating NADH the rate of formation is substantially smaller than that with succinate as the substrate. In contrast to NADH oxidase,the rate-substrate concentration dependence for the superoxide production shows a maximum at low (50 µM)concentrations of NADH. NAD⁺ and NADH inhibit the succinate-supported superoxide generation. Deactivation of Complex I results in almost complete loss of its NADH-ubiquinone reductase activity and in increase in NADH-dependent superoxide generation. A model is proposed according to which complex I has two redox active nucleotide binding sites.One site (F) serves as an entry for the NADH oxidation and the other one (R) serves as an exit during either the succinate-supported NAD⁺ reduction or superoxide generation or NADH-ferricyanide reductase reaction.Translated from Biokhimiya, Vol. 70, No. 2, 2005, pp. 150–159.Original Russian Text Copyright © 2005 by Vinogradov, Grivennikova.This revised version was published online in April 2005 with corrections to the post codes.     

Kinetic Modeling of Energy Metabolism and Superoxide Generation in Hepatocyte Mitochondria

Direct nonenzymatic oxidation of semiquinone by oxygen is one of the main sources of superoxide radicals in mitochondria. Using all the known data on hepatocyte mitochondria, we have revealed the correlation between the rate of superoxide generation by the bc ₁complex and the transmembrane potential (). Assuming that the main electrogenic stage of the Qcycle is the electron transfer between the cytochrome bhemes, then the rate of superoxide generation sharply increases when grows from 150 to 180 mV. However, this interrelation is ambiguous. Indeed, the increase of the generation rate with the growth of the potential can occur faster when succinate dehydrogenase is inhibited by malonate than when external ADP is exhausted. When the potential is changed by adding phosphate or potassium (K⁺), the rate of production remains constant, although the comparison of the rates at the same reveals the effect of phosphate or potassium. It turned out that the rate of generation is a function of rather than any of its components. Phosphate and K⁺have practically no influence on , since the change in is compensated by pH. The rate of superoxide generation by the bc ₁complex is a multiple function of the electron-transfer activity of enzymes, the processes determining the membrane potential (e.g., loading), and the oxygen concentration. The kinetic model proposed in this work may serve to understand how the superoxide production is regulated.     

Characterization of microbial NO production, N2O production and CH4 oxidation initiated by aeration of anoxic rice field soil

Intermittent drainage of rice fields isdiscussed as an option to mitigate emission ofCH₄, an important greenhouse gas. HoweverN₂O, a potentially more effective greenhouse gas,may be emitted during the aeration phase. Therefore,the metabolism of NO, N₂O, NH ,NO and NO and the kinetics ofCH₄ oxidation were measured after aeration ofmethanogenic rice field soil. Before aeration, thesoil contained NH in relatively highconcentrations (about 4 mM), while NO andNO were almost undetectable. Immediatelyafter aeration both NO and N₂O were produced withrates of about 15 pmol h^-1 gdw^-1 and 5 pmolh^-1 gdw^-1, respectively. Simultaneously,NH decreased while NO accumulated. Later on, NO was depletedwhile NO concentrations increased.Characteristic phases of nitrogen turnover wereassociated with the activities of ammonium oxidizers,nitrite oxidizers and denitrifiers. Oxidation ofNH and production of NO and N₂O wereinhibited by 10 Pa acetylene demonstrating thatnitrification was obligatory for the initiation ofnitrogen turnover and production of NO and N₂O.Ammonium oxidation was not limited by the availableNH and thus, concomittant production of NOand N₂O was not stimulated by addition ofNH . However, addition of NO stimulated production of NO and N₂O in bothanoxic and aerated rice soil slurries. In this case,10 Pa acetylene did not inhibit the production of NOand N₂O demonstrating that it was due todenitrification which was obviously limited by theavailability of NO . In the aerated soilslurries CH₄ was only oxidized if present atelevated concentrations >50 ppmv CH₄). Atatmospheric CH₄ concentrations (1.7 ppmv)CH₄ was not consumed, but was even slightly produced.CH₄ oxidation activity increased afterpreincubation at 20% CH₄, and then CH₄was also oxidized at atmospheric concentrations. CH₄oxidation kinetics exhibited sigmoid characteristicsat low CH₄ concentrations presumably because ofinhibition of CH₄ oxidation by NH .     

Growth yields and saturation constant of Desulfovibrio vulgaris in chemostat culture

Desulfovibrio vulgaris (strain Marburg) was grown on H₂ and sulfate as sole energy source in a chemostat limited by the sulfate supply. The biomass concentration and the sulfate concentration in the culture were determined as a function of the dilution rate. From the data a K _S (saturation constant) for sulfate of 10 M, a _max of 0.23 h^–1, and a of 13 g/mol were calculated. The organism was also grown in chemostat culture on H₂ and sulfite, H₂ and thiosulfate, and pyruvate (without sulfate). was found to be 35 g/mol, 36 g/mol, and Y _pyr ^max 10 g/mol. The growth yields are discussed with respect to ATP gains in dissimilatory sulfate reduction.     

Protective Effect of Rare Earth Against Oxidative Stress Under Ultraviolet-B Radiation

The effects of lanthanum (III) (La(III)) in protecting soybean leaves against oxidative stress induced by ultraviolet-B (UV-B) radiation were investigated. The increase in contents of hydrogen peroxide (H₂O₂) and superoxide () due to UV-B radiation suggested oxidative stress. The increase in the content of malondialdehyde (MDA) and the decrease in the index of unsaturated fatty acid (IUFA) indicated oxidative damage on cell membrane induced by UV-B radiation. La(III) partially reversed UV-B-radiation-induced damage of plant growth. The reduction in the contents of H₂O₂, , and MDA and increase in the content of IUFA, compared with UV-B treatment, also indicated that La(III) alleviated the oxidative damage induced by UV-B radiation. The increase in the activities of superoxide dismutase and peroxidase and the contents of ascorbate, carotinoids, and flavonoids were observed in soybean leaves with La(III) + UV-B treatment, compared with UV-B treatment. Our data suggested that La(III) could protect soybean plants from UV-B-radiation-induced oxidative stress by reacting with reactive oxygen species directly or by improving the defense system of plants.     

Minimal Functional Unit of Lactate Dehydrogenase

The tetrameric heart isozyme of lactate dehydrogenase (H₄) is modified by p-chloromercuribenzoate (PCMB) to produce the inactive tetramer and then hybridized with native tetrameric muscle isozyme (M₄). The hybrid mixture was isolated by polyacrylamide gel electrophoresis (PAGE) and then stained for enzyme activity and with Coomassie brilliant blue. Only three bands were found on the gels in either case. The hybrid enzymes as isolated by PAGE have half the specific activity of the native muscle enzyme. The electrophoresis properties of HM₃ are very similar to those of HM₃, while the electrophoresis properties of are very similar to those of H₂M₂. The above results strongly suggest that the tetramer having enzymatic activity contains at least two native subunits, and the di-subunit in the tetrameric enzyme is the minimal functional unit.     

Physiological mechanisms for underwater endurance: Canada goose (Branta canadensis) versus Pekin duck (Anas platyrhynchos)

Maximum submergence time of Canada geese was 18% of that of similarly sized Pekin ducks. Due to a smaller respiratory system volume the oxygen store of Canada geese was 82% of that of Pekin ducks, accounting for approximately 33% of the difference in underwater survival times. The respiratory properties and volume of the blood were similar in both species. Both species utilised approximately 79% of the respiratory oxygen store and 90% of the blood oxygen store. Therefore, most of the species difference in survival times was due to a less effective oxygen-conserving cardiovascular response (bradycardia, peripheral vasoconstriction) in Canada geese. Duck cardiac chronotropic sensitivity to hypoxia during submergence was twice that observed in geese. Furthermore, a lower hypoxic ventilatory response was observed in geese than in ducks. Density of monoamine varicosities in hindlimb artery walls was lower in geese than ducks. However, electrical stimulation of the hindlimb muscles did not cause ascending vasodilation during submergence in either species, perhaps due to higher levels of catecholamines in submerged geese. We conclude that the major difference between species is higher oxygen chemosensitivity in ducks which effects a much more rapid and efficacious oxygen-conserving response during forced submergence.Abbreviations ATPS · BTPS · STPD CNS central nervous system - EEG electroencephalogram - ECG electrocardiogram - EDTA ethylenediaminetetra-acetic acid - HPLC high performance liquid chromatography - fractional oxygen concentration of inspired air - pre-immersion fractional concentration of oxygen in the respiratory system - pre-emersion fractional concentration of oxygen in the respiratory system - [Hb] haemoglobin concentration - Hct haematocrit - HR heart rate - M _B body mass - M _b brain mass - M _h heart mass - partial pressure of carbon dioxide in arterial blood - partial pressure of oxygen in arterial blood - SPG sucrose-potassium phosphate-glyoxylic acid - t _d maximum underwater survival time - respiratory minute volume - V _pl plasma volume - V _rs respiratory system volume - accessible respiratory system oxygen store - total non-myoglobin-bound oxygen store - V _tb blood volume - blood oxygen store     

Phytotoxicity of aluminium-fluoride complexes and their uptake from solution culture by Avena sativa and Lycopersicon esculentum

Avena sativa (oats) and Lycopersiconesculentum (tomatoes) were grown in dilute nutrientsolutions supplemented with Al only, F only or acombination of both. In solutions containing Al andF, shoot growth was limited when predicted Al_r(Al³⁺, AlOH²⁺ and Al(OH) )activities were < 0.1 M, activities three orders ofmagnitude lower than the critical value determinedwith Al only. The data suggest that at the activitiesused in these experiments, Al_r is most toxic,AlF²⁺ and AlF are toxic to a lesserextent, and AlF₃, AlF and F^-are least toxic. Fluoride concentrations in shootsgrown in solutions correlated best with positivelycharged AlF species (i.e.AlF²⁺, AlF ) and the molar ratio ofF:Al in most plants shoots was about 3:2. However,when activities of positively chargedAlF species were low (< 50 M)and theactivity of AlF₃ species high (500 M) the molarratio of F:Al in plant shoots suggested AlF₃ wastaken up. These findings are discussed in relation toplant uptake and toxicity. Measured concentrations of Al_r and F^- insolutions containing Al and F were compared withconcentrations predicted by a computer model(GEOCHEM-PC). The method for measuring F^-concentrations, using a non-complexing buffer and Fion-selective electrode, gave good agreement withpredicted F^- concentrations. The8-hydroxyquinoline method for measuring Al_rconcentrations did not agree with predictedconcentrations, highlighting the limitations of thismethod when measuring Al_r in the presence of F ina multi-ligand system with high concentrations of Fand Al.     

High groundwater nitrate concentrations inhibit eutrophication of sulphate-rich freshwater wetlands

During the last 60 years, pollution of the groundwater with has greatly increased in many parts of Europe, as a consequence of excessive use of manure and synthetic fertilisers. Monitoring of groundwater-fed wetlands indicated that sediments with high concentrations had the lowest Fe and concentrations in the pore water. A comparison of two restored open water fens, differing in supply via the groundwater, indicated that the redox potential and the sulphate ( ) reduction rate were lower when the groundwater contained not only but also high concentrations. The lower reduction rates in the -rich open water fen were associated with lower concentrations and the presence of plant species characteristic of clear water. In contrast, the higher reduction rates in the -poor open water fen were associated with very high concentrations and massive development of plant species characteristic of eutrophic environments. Investigations at -rich seepage sites in black alder carrs, showed that high concentrations in the pore water caused chlorosis in the alder carr vegetation, due to lower availability of Fe in the pore water and less Fe uptake by the plants. Experimental desiccation of sediments proved that the -rich seepage sites contained no oxidisable FeS _x , contrary to -poor locations, which became acidified and mobilised extremely high amounts of due to FeS _x oxidation. A laboratory experiment showed that addition to sediments led to reduced releases of Fe, and S^2–, very likely due to the oxidation of reduced Fe and S compounds. Overall, the results confirmed that is an energetically more favourable electron acceptor in anaerobic sediments than Fe and , and that high loads function as a redox buffer, preventing reduction of Fe and . Limited reduction prevents S^2– -mediated mobilisation of from Fe- complexes. At a higher redox potential, reduced Fe, including FeS _x , was oxidised, increasing the content of Fe(III) capable of binding . This prevented increased availability and the concomitant massive development of plant species characteristic of eutrophic environments.     

A comparative study of the mechanism of Na+ and Cl? excretion by the gill ofMugil capito andFundulus heteroclitus: Effects of Stress

Summary In seawater (SW)-adaptedMugil andFundulus, gill effluxes of Na⁺ and of Cl^– and the simultaneously recorded transgill potential (P.D.) differ according to whether they are measured in stressed or rested animals.In rested animals of the two species, transfer to Ringer's solution considerably reduces the P.D. but not . InFundulus, is also decreased. Transfer of the two species from SW to fresh water (FW) reduces and by 75 to 85% and leads to a large inversion of P.D. When K⁺ is added to FW, a gill depolarization occurs, as well as a large increase of and .These results suggest that: 1) the P.D. originates primarily from the diffusion of cations, the gill permeability to Na⁺ ( ) being greater than that to Cl^– ( ), 2) a Cl^–/Cl^– exchange independent of P.D. is associated with the Cl^– pump; 3) Cl^– pump activity is linked to Na⁺/K⁺ exchange which in turn is associated to a Na⁺/Na⁺ exchange diffusion mechanism.In stressed individuals of the two species, the P.D. in SW, as well as the P.D. changes observed during transfer experiments, are considerably reduced. The decrease of and observed after transfer from SW to FW are also minimised. Changes are smaller inFundulus. The decrease of P.D. characterizing stressed animals may be at least in part due to a 3 to 4 fold increase of which becomes equal to in both species.As a result of stress, the K⁺-activated Na⁺ and Cl^– excretion mechanisms are totally inhibited inFundulus and partially so inMugil.Stress response seems more intense inFundulus and recovery from stress faster inMugil.     

Antioxidant defense responses: physiological plasticity in higher plants under abiotic constraints

Environmental stresses (salinity, drought, heat/cold, light and other hostile conditions) may trigger in plants oxidative stress, generating the formation of reactive oxygen species (ROS). These species are partially reduced or activated derivatives of oxygen, comprising both free radical and non-radical (H₂O₂) forms, leading to cellular damage, metabolic disorders and senescence processes. In order to overcome oxidative stress, plants have developed two main antioxidants defense mechanisms that can be classified as non-enzymatic and enzymatic systems. The first class (non-enzymatic) consists of small molecules such as vitamin (A, C and E), glutathione, carotenoids and phenolics that can react directly with the ROS by scavenging them. Second class is represented by enzymes among them superoxide dismutase, peroxidase and catalase which have the capacity to eliminate superoxide and hydrogen peroxide. In this review, we have tried to explore the related works, which have revealed the changes in the basic antioxidant metabolism of plants under various abiotic constraints.     

Larmor frequency selective model free analysis of protein NMR relaxation

Summary The Lipari-Szabo dynamical formalism is extended by setting the time constants of the Lorentzian terms to and . This analysis is compared to the earlier proposed three-parameter extended model free formalism with regard to the range of equivalence and the advantages of the simplified two-parameter (S _inff ^sup2 ,S _infH ^sup2 ) and (S _inff ^sup2 ,S _infN ^sup2 ) representations. Spectral density components are calculated and compared to those obtained from the spectral density analysis formalism. Protein relaxation data, commonly analyzed in terms of the two-parameter representation, may correspond to a dynamically heterogeneous behaviour that is more appropriately represented in terms of a fast limit order parameter and a second, lower frequency order parameter.     

ESR-Untersuchungen zur Radikalstruktur in bestrahltem L-Leucinhydrochlorid

Zusammenfassung Bei Röntgenbestrahlung von L-Leucin·HCl bei Zimmertemperatur wird das >C-H-Proton unter Bildung des Radikals (CH₃)₂ C-CH₂-CHNH₃ ⁺-COOH abgetrennt. Das von acht -Protonen stammende ESR-Spektrum wurde wegen nur schwacher Anisotropie der -Protonen-Hfs auch im polykristallinen Zustand vollständig aufgelöst, und es wurden folgende isotrope Aufspaltungen ermittelt: bei Messung bei Zimmertemperatur = 23.5±1 Gauß für sechs äquivalente CH₃-Protonen und =8,0±1 Gauß und = 38,8±1 Gauß für die zwei Methylenprotonen, bei Messung bei 77 °K entsprechend = 22,6±1 Gauß =8,8±1 Gauß und =45,2±1 Gauß (Temperaturabhängigkeit der CH₂-Protonenkopplung). Das bei Zimmertemperatur beobachtete Radikal wird bereits durch Bestrahlung bei 77 °K gebildet.

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ESR-studies on the structure of radicals in X-irradiated L-leucine hydrochloride

Summary The structure of radicals in X-irradiated polycrystalline L-leucine·HCl has been studied by ESR. At room temperature the radical (CH₃)₂ > C-CH₂-CHNH ₃ ⁺ -COOH is formed by abstraction of the >C-H-proton. The isotropic part of hyperfine interaction with eight -protons has been measured even in polycrystalline state with following values: at 300 °K =23,5±1 gauss for six equivalent CH₃-protons and =8.0±1 gauss and =38.8±1 gauss for two CH₂-protons, at 77 °K =22.6±1 gauss, =8.8±1 gauss and =45-2±1 gauss respectively (the coupling of the CH₂-protons is dependent from temperature). This radical is present already following irradiation at 77 °K.

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Herrn Prof. Dr. habil. Kh.Lohs möchten wir an dieser Stelle für die Unterstützung dieser Arbeit und wertvolle Diskussionen danken. Herrn Ing. N.Klimes, Frl. G.Klein und Herrn J.Benkert gilt unser Dank für die technische Assistenz, Herrn Ing. M.Kresse für die Bestrahlung der verwendeten Proben.     

Biological functions of antioxidants in plant transformation

Browning and necrosis of transformed cells/tissues, and difficulty to regenerate transgenic plants from the transformed cells/tissues (recalcitrance) are common in Agrobacterium-mediated transformation process in many plant species. In addition, most crop transformation methods that use NPTII selection produce a significant number of nontransgenic shoots, called “shoot escapes” even under stringent selection conditions. These common problems of plant transformation, (browning and necrosis of transformed cells/tissues, recalcitrance, and the occurrence of shoot escapes) severely reduces transformation efficiency. Recent research indicates that reactive oxygen species (ROS) such as superoxide radical , the hydrogen peroxide (H₂O₂), the hydroxyl radical (OH′), and the peroxyl radical () may be playing an important role in tissue browning and necrosis during transformation. This review examines the role of ROS in in vitro recalcitrance and genetic transformation and the opportunities to improve transformation efficiency using antioxidants.     

Influence of hyperbaric oxygen tension on the metabolism of Candida tropicalis and Rhodococcus erythropolis

Summary The effect on metabolism of hyperbaric dissolved oxygen tension in batch cultures of Candida tropicalis (Cast.) Berkhout and Rhodococcus erythropolis with three different carbon sources was studied in a 20-l bioreactor under controlled conditions. The respiratory quotient was not significantly influenced by dissolved oxygen concentrations up to 40 mg/l oxygen. Elementary cell composition and proportional contents of DNA, RNA, and protein were not markedly influenced by the various oxygen concentrations but depended mainly on the growth rate. It was found that the production of trehalose lipid by R. erythropolis was dependent on the growth rate which could be enhanced by raising the oxygen concentration. The specific activity of catalase was affected more by the nature of the carbon source than by increased oxygen concentration. On the basis of the experimental data the application of oxygen-enriched air for biotechnological processes is discussed.Symbols and abbreviations k_La Specific volumetric oxygen transfer rate - Oxygen consumption rate, grams oxygen per hour and per liter - Carbon dioxide production rate, grams carbon dioxide per hour and per liter - RQ Respiratory quotient, - t Cultivation time - Y_X/S Yield coefficient, grams cell dry weight/grams substrate - Yield coefficient, grams cell dry weight/grams oxygen consumed - Y_kJ Yield coefficient, grams cell dry weight/heat of combustion of the consumed substrate - Y_ave– Yield coefficient, grams cell dry weight/mol available electrons of the consumed substrate     

Ventilation and control of acid-base status during temperature acclimation in the crab,Cancer magister

Summary Frequencies of scaphognathite (ventilatory,f _sc) and heart (f _h) pumping, oxygen consumption ( ), and hemolymph oxygen, carbon dioxide and pH levels were measured in adult Dungeness crabs (Cancer magister) during 7–10 day periods of exposure to 7, 12, and 17°C seawater. Ventilation volume ( ) was calculated for individual animals fromf _sc and a previously determined relationship between stroke volume and animal mass. increases (Q₁₀=2.3) with temperature were associated with larger increases inf _sc (Q₁₀=3.3) and (Q₁₀=3.5) and smaller increases inf _h (Q₁₀=1.5). The incidence of unilateral scaphognathite pumping and pausing decreased as temperature rose.Postbranchial oxygen tension was maintained in vivo but hemolymph oxygen content decreased both in vivo and in vitro as temperature rose. Postbranchial carbon dioxide tension did not change significantly but relative alkalinity was maintained as temperature rose by loss of hemolymph bicarbonate. The effects of increased ventilation volume and potential mechanisms of bicarbonate regulation are discussed.The responses of the essentially subtidalCancer magister are compared with those of subtidal, intertidal and terrestrial crabs demonstrating that the concepts of acid-base regulation developed for water and air breathing vertebrates are also applicable to water and air breathing crabs, and that intertidal crabs may exhibit transitional states.This work was supported by Grant No. A.5762 National Research Council of Canada