The respiration of young coho Oncorhynchus kisutch at gradually declining oxygen levels and during recovery from oxygen deprivation

The respiration of coho salmon, Oncorhynchus kisutch , weighing between 15 and 50 g was measured at gradually declining oxygen levels and at temperatures ranging between 14 and 17°C. The maximum and minimum oxygen concentrations tested were 250 and 40 μmol L⁻¹, respectively. Respiration rates were measured for 1 h periods before oxygen concentration was lowered by 12.5 or 25.0 μmol oxygen L⁻¹. At the end of these endurance tests the oxygen level was returned to normoxic conditions and respiration rates were determined for the recovery period. Under normoxic conditions (> 200 μmol L⁻¹) the respiration of coho levelled around 5.1 μmol g⁻¹ wet weight h⁻¹. At intermediate levels between 150 and 200 μmol oxygen L⁻¹, the average rate increased to 5.8 μmol g⁻¹ h⁻¹, which could be attributed to higher spontaneous activity of the test animals. At low oxygen levels (< 150 μmol⁻¹) average respiration rates dropped to values between 5.5 and 5.7 μmol g⁻¹ h⁻¹, reaching a minimum of 3.8 μmol g⁻¹ h⁻¹ at oxygen levels below 50 μmol L^μ. First mortality was observed in this range. After exposure to reduced oxygen levels the fish maintained a higher respiration rate when again exposed to normoxic oxygen levels above 200 μmol L⁻¹. Increased respiration rates were observed for a recovery period of 6 h.     

Individual variation in the rate of oxygen consumption by zebrafish embryos

A sensitive microsensor‐based method was used to measure oxygen consumption of individual zebrafish Danio rerio embryos at 6 h intervals from 24 to 75 h post‐fertilization. An increase in oxygen consumption rates from 4·54 to 8·29 nmol O₂ h⁻¹ was found during this period. At the individual level the differences in oxygen consumption rates caused the total oxygen consumption from 24 to 75 h post‐fertilization to vary between 0·261 and 0·462 μmol O₂ per individual with a mean of 0·379 μmol O₂ per individual. A separate carbon mass balance study corroborated the mean total oxygen consumption obtained by yielding a respiratory quotient of 0·80 for this period. These results suggest that there is significant intraspecific variation in the metabolic rate of developing zebrafish embryos, which may influence other early life‐history traits such as growth and starvation resistance.     

Understanding your inhibitions: effects of GABA and GABAA receptor modulation on brain cortical metabolism

A targeted neuropharmacological, ¹H/¹³C NMR spectroscopy and multivariate statistical approach was used to examine the effects of exogenous GABA and ligands at the GABA_A receptor family on brain metabolism in the Guinea pig cortical tissue slice. All ligands at GABA_A receptors generated metabolic patterns which were distinct from one another with the major variance in the data arising because of metabolic work (shown by net flux into Krebs cycle byproducts and increased metabolic pool sizes). Three major clusters of metabolic signatures were identified which corresponded to: (i) activity at phasic (synaptic) GABA_A receptors, dominated by α1-containing receptors and responsive to GABA at 10 μmol/L; (ii) activity at perisynaptic receptors, dominated by response to high (40 μmol/L) GABA and the superagonist 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridine-3-ol hydrochloride, and C, activity at extrasynaptic receptors, dominated by response to low (0.1–1.0 μmol/L) GABA, zolpidem (400 nmol/L) and the non-specific allosteric modulator RO19-4603 (1 nmol/L). These results highlight the utility of a different but robust approach to study of the GABAergic system using metabolic systems analysis.     

Hydroxytyrosol protects retinal pigment epithelial cells from acrolein-induced oxidative stress and mitochondrial dysfunction

Hydroxytyrosol (HTS) is a natural polyphenol abundant in olive oil. Increasing evidence indicates HTS has beneficial effect on human health for preventing various diseases. In the present study, we investigated the protective effects of HTS on acrolein-induced toxicity in human retinal pigment epithelial cell line, ARPE-19, a cellular model of smoking- and age-related macular degeneration. Acrolein, a major component of the gas phase cigarette smoke and also a product of lipid peroxidation in vivo , at 75 μmol/L for 24 h caused significant loss of cell viability, oxidative damage (increase in oxidant generation and oxidative damage to proteins and DNA, decrease in antioxidants and antioxidant enzymes, and also inactivation of the Keap1/Nrf2 pathway), and mitochondrial dysfunction (decrease in membrane potential, activities of mitochondrial complexes, viable mitochondria, oxygen consumption, and factors for mitochondrial biogenesis, and increase in calcium). Pre-treatment with HTS dose dependently and also time dependently protected the ARPE-19 cells from acrolein-induced oxidative damage and mitochondrial dysfunction. A short-term pre-treatment with HTS (48 h) required > 75 μmol/L for showing protection while a long-term pre-treatment (7 days) showed protective effect from 5 μmol/L on. The protective effect of HTS in this model was as potent as that of established mitochondria-targeting antioxidant nutrients. These results suggest that HTS is also a mitochondrial-targeting antioxidant nutrient and that dietary administration of HTS may be an effective measure in reducing and or preventing cigarette smoke-induced or age-related retinal pigment epithelial degeneration, such as age-associated macular degeneration.     

A method for long-term measurement of respiration in intertidal fishes during simulated intertidal conditions

Aquatic and aerial respiration of the amphibious fishes Lipophrys pholis and Periophthalmus barbarus were examined using a newly designed flow-through respirometer system. The system allowed long-term measurements of oxygen consumption and carbon dioxide release during periods of aquatic and aerial respiration. The M o ₂ of L. pholis , measured at 15° C, was 2·1 μmol O₂ g^–1 h^–1 during aquatic and 1·99 μmol O₂ g^–1 h^–1 during aerial exposure. The corresponding values of the M co₂ were 1.67 and 1.59 μmol O₂ g^–1 h^–1 respectively, giving an aquatic respiratory exchange ratio (RER) of 0·80 and an aerial RER of 0·79. The M o₂ of P. barbarus , measured at 28°C, was 4·05 μmol O₂ g^–1 h^–1 during aquatic and 3·44 μmol O₂ g^–1 h^–1 during aerial exposure. The corresponding values of the Mco₂ were 3·29 μmol CO₂ g^–1 h^–1 and 2·63 μmol CO₂ g^–1 h^–1 respectively, giving an aquatic RER of 0·81 and an aerial RER of 0·77. While exposed to air for at least 10 h, both species showed no decrease in metabolic rate or carbon dioxide release. The RER of these fishes equalled their respiratory quotient. After re-immersion an increased oxygen consumption, due to the payment of an oxygen debt, could not be detected.     

Measurement of guard-cell respiration rates using Cartesian-diver technique

Dark respiration rates of guard-cell protoplasts of Commelina communis L. were measured over a temperature range (15–30° C) using a Cartesian-diver microrespirometry technique. Measurements were made using a few microliters of suspension medium containing between 400 and 3 700 protoplasts. Respiration rates were approximately linear for at least 1 h at all temperatures. Respiration rates increased rapidly between 20 and 25° C to relatively high levels (6.11·10^-6 mol O₂ h^-1 protoplast^-1=1259 mol O₂ mg^-1 chlorophyll h^-1=22.97 mol O₂ mg^-1 protein h^-1) with no further increases above this temperature. Respiration rates were much lower in protoplasts 15–16 h old than in freshly prepared ones indicating considerable deterioration of their viability over this time period.     

Enhancement of Bacillus thuringiensis with monosodium glutamate against larvae of obliquebanded leafroller (Lep.:Tortricidae)

Abstract:  Monosodium glutamate, a taste enhancer widely used in food industry, was tested in the laboratory to determine its phagostimulatory effects on larvae of Choristoneura rosaceana (Harris). Larvae fed apple leaves treated with 50–700  μ g/l monosodium glutamate increased leaf tissue consumption by approximately 40%. The stimulatory effect of monosodium glutamate (at 675  μ g/l concentration) was maintained throughout 10 days of continuous exposure. Adding 675  μ g/l monosodium glutamate to commercial formulation of Bacillus thuringiensis ssp. kurstaki, (DiPel^®2X DF), lowered LC₅₀ from 450 to 150  μ g/l (P < 0.05, Lethal Ratio Significance Test), indicating good potential for monosodium glutamate to enhance B. thuringiensis -based formulations.     

Characterization of the submerged growth ofMoraxella bovis

A submerged batch cultivation ofMoraxella bovis in a medium containing enzymic casein hydrolyzate and supplemented with dialyzed ram blood was described. Up to the growth limitation the bacteria grew exponentially without a lag phase and with a doubling period of 64 min. During the exponential phase no significant decrease of viable cells and cell autolysis were observed. Amino acids were the limiting factor. At the end of growth glutamie acid, threonine and serine were detected at the lowest concentrations. Substrate limitation led to an irreversible decrease of the metabolic activity. Growth yield on oxygen was 2.3 × 10⁹ cells per mg O₂. The fraction of respiration required for growth-uncoupled processes was negligible. During the cultivation ammonia was produced and the pH increased but it was not the inhibitory factor. Respiration was not limited by oxygen at concentrations higher than 0.63 μmol O₂ per L. Sufficient pili were produced.     

PHYSIOLOGICAL RESPONSES TO TEMPERATURE AND IRRADIANCE IN SPIROGYRA (ZYGNEMATALES, CHAROPHYCEAE)1

Spirogyra Link (1820) is an anabranched filamentous green alga that forms free-floating mats in shallow waters. It occurs widely in static waters such as ponds and ditches, sheltered littoral areas of lakes, and stow-flowing streams. Field observations of its seasonal distribution suggest that the 70-μm-wide filament form of Spirogyra should have a cool temperature and high irradiance optimum for net photosynthesis. Measurements of net photosynthesis and respiration were marie at 58 combinations of tight and temperature in a controlled environment facility. Optimum conditions were 25°C and 1500 μmol photons m⁻² s⁻¹, at which net photosynthesis averaged 75.7 mg O₂ gdm⁻¹ h⁻¹. Net photosynthesis was positive at temperatures from 5° to 35°C at most irradiances except at combinations of extremely low irradiances and high temperatures (7 and 23 μmol photons m⁻² s⁻¹ at 30°C and 7, 23, and 35 μmol photons m⁻² s⁻¹ at 35°C). Respiration rates increased with both temperature and prior irradiance. Light-enhanced respiration rates were significantly greater than dark respiration rates following irradiances of 750 μmol photons m⁻² s⁻¹ or greater. Polynomials were fitted to the data to generate response surfaces; such response surfaces can be used to represent net photosynthesis and respiration in ecological models. The data indicate that the alga can tolerate the cool water and high irradiances characteristic of early spring but cannot maintain positive net photosynthesis under conditions of high temperature and low light (e.g. when exposed to self-shading ).     

Hydralazine inhibits compression and acrolein-mediated injuries in ex vivo spinal cord

We have previously shown that acrolein, a lipid peroxidation byproduct, is significantly increased following spinal cord injury in vivo , and that exposure to neuronal cells results in oxidative stress, mitochondrial dysfunction, increased membrane permeability, impaired axonal conductivity, and eventually cell death. Acrolein thus may be a key player in the pathogenesis of spinal cord injury, where lipid peroxidation is known to be involved. The current study demonstrates that the acrolein scavenger hydralazine protects against not only acrolein-mediated injury, but also compression in guinea pig spinal cord ex vivo . Specifically, hydralazine (500 μmol/L to 1 mmol/L) can significantly alleviate acrolein (100–500 μmol/L)-induced superoxide production, glutathione depletion, mitochondrial dysfunction, loss of membrane integrity, and reduced compound action potential conduction. Additionally, 500 μmol/L hydralazine significantly attenuated compression-mediated membrane disruptions at 2 and 3 h following injury. This was consistent with our findings that acrolein-lys adducts were increased following compression injury ex vivo , an effect that was prevented by hydralazine treatment. These findings provide further evidence for the role of acrolein in spinal cord injury, and suggest that acrolein-scavenging drugs such as hydralazine may represent a novel therapy to effectively reduce oxidative stress in disorders such as spinal cord injury and neurodegenerative diseases, where oxidative stress is known to play a role.     

Artificial capillary perfusion cell culture: Metabolic studies

Summary Glucose, lactic-acid, and oxygen metabolism of BHK and L929 cells on artificial capillary perfusion units have been studied using several different modes of perfusion. After 7 to 10 days, cells planted in the extracapillary compartment of culture units containing 80 to 150 fibers reached populations that used 0.073±0.025 μmol per min glucose and 0.76±0.26 μl per min oxygen and excreted 0.078±0.038 μmol per min lactic acid. From these data it is estimated that these units contain approximately 2×10⁷ cells. The metabolic rate of cultures perfused through the capillaries or through the extracapillary compartment was not affected significantly by change in flow rate except at perfusion flow rates ≤0.05 ml per min. The cell population, as measured by metabolic activity, did not increase significantly when the serum content of the medium was ≤1%. No major differences were found in glucose utilization rates of equal numbers of cells on artificial capillaries, on short-term suspension culture, or as monolayers in plastic flasks. Artificial capillary perfusion may provide a simple system for studying metabolism of mammalian cells in culture. Research was supported by the U.S. Army Medical Research and Development Command, Washington, D.C. 20314, under Contract No. DAMD 17-76-C-5075.     

Effects of Irradiance and Temperature on the Photosynthesis and Vegetative Propagation of Caulerpa serrulata

The photosynthetic oxygen evolution of Caulerpa serrulata was determined with oxygen electrodes. The effects of light and temperature on the growth and regeneration of fragmented C. serrulata thalli were analyzed. The regenerating rate and establishment of different sizes and portions of C. serrulata were studied. The results showed that the light saturation point of C. serrulata was 200 μmol photons/m2 per s and the optimum growth temperature was 25-30℃. Under these conditions, the maximum photosynthetic oxygen evolution rate was 15.1:±0.29 mg O2/mg Chl a/h, the growth rate and elongation rate reached the highest values, 4.67±0.09 mg FW/d and 0.78±0.01 mm/d, respectively. The fragmented C. serrulata thalli was regenerated at 20-35℃ and survived at 15℃ and 200 μmnol photons/m2 per s. A different survival rate was detected according to fragment size. All of these results indicated that C. serrulata was a candidate to become an invasive species if introduced into a new place. Therefore, we should pay more attention to C. serrulata for its potential threat to marine ecosystem when it is sold for aquarium use.     

Antarctic stream ecosystems: physiological ecology of a blue-green algal epilithon

SUMMARY. 1. Several dozen summer meltwater streams are located in the McMurdo Sound region (c. 78°S 165°E) of southern Victoria Land. They are characterized by a highly variable flow regime at diel, seasonal and annual times caleis; wide fluctuations in temperature and nutrient content; and a very simple epilithic community of cyanophytes ( Nostoc spp., Oscillatoriaceae), bacteria, fungi and microherbivores.
2. The epilithon survives the dark Antarctic winter as dry, frozen mats which provide a large inoculum for growth the following summer. This overwintering assemblage retains a high metabolic capacity and responds rapidly to rehydration.
3. In a series of artificial substrate experiments, biomass accumulation rates were generally less than 0.1 In units d⁻¹. Colonization and growth on the substrates was inversely related to the suspended sediment load of the stream. There was also a visual correspondence between per cent algal cover of the natural streambed and the clarity of the streamwater. Sloughing losses may limit community biomass, particularly in the turbid flowing waters.
4. During running water conditions the mature communities had very low gross photosynthetic rates per unit chlorophyll (<0,1 μg C (μg chl a .h)⁻¹ and per unit carbon (<0,2 μg C (mg biomass C.h)⁻¹). Respiration was generally a high percentage (up to 92%) of gross photosynthesis, which probably reflected the high population densities of microheterotrophs in the community.
5. The floristically simple epilithic mats slowly accumulate to extreme biomass levels (>20 μg chl a cm ⁻², <20 mg C cm⁻²). Production rates per unit biomass are low, probably in response to the cold temperatures of the Antarctic stream environment, and the accumulated biomass represents several seasons of growth.     

Lactate uptake contributes to the NAD(P)H biphasic response and tissue oxygen response during synaptic stimulation in area CA1 of rat hippocampal slices

Synaptic train stimulation (10 Hz × 25 s) in hippocampal slices results in a biphasic response of NAD(P)H fluorescence indicating a transient oxidation followed by a prolonged reduction. The response is accompanied by a transient tissue PO₂ decrease indicating enhanced oxygen utilization. The activation of mitochondrial metabolism and/or glycolysis may contribute to the secondary NAD(P)H peak. We investigated whether extracellular lactate uptake via monocarboxylate transporters (MCTs) contributes to the generation of the NAD(P)H response during neuronal activation. We measured the effect of lactate uptake inhibition [using the MCT inhibitor α-cyano-4-hydroxycinnamate (4-CIN)] on the NAD(P)H biphasic response, tissue PO₂ response, and field excitatory post-synaptic potential in hippocampal slices during synaptic stimulation in area CA1 (stratum radiatum). The application of 4-CIN (150–250 μmol/L) significantly decreased the reduction phase of the NAD(P)H response. When slices were supplemented with 20 mmol/L lactate in 150–250 μmol/L 4-CIN, the secondary NAD(P)H peak was restored; whereas 20 mmol/L pyruvate supplementation did not produce a recovery. Similarly, the tissue PO₂ response was decreased by MCT inhibition; 20 mmol/L lactate restored this response to control levels at all 4-CIN concentrations. These results indicate that lactate uptake via MCTs contributes significantly to energy metabolism in brain tissue and to the generation of the delayed NAD(P)H peak after synaptic stimulation.     

A critical evaluation of a flow‐cell based on a liquid core waveguide for chemiluminescence measurements

Liquid‐core waveguides (LCWs), devices that constrain the emitted radiation minimizing losses during the transport, are an alternative to maximize the amount of detected radiation in luminescence. In this work, the performance of a LCW flow‐cell was critically evaluated for chemiluminescence measurements, by using as model the oxidation of luminol by hydrogen peroxide or hypochlorite. An analytical procedure for hypochlorite determination was also developed, with linear response in the range 0.2–3.8 mg/L (2.7–51 µmol/L), a detection limit estimated as 8 µg/L (0.64 µmol/L) at the 99.7% confidence level and luminol consumption of 50 µg/determination. The coefficients of variation were 3.3% and 1.6% for 0.4 and 1.9 mg/L ClO^?, respectively, with a sampling rate of 164 determinations/h. The procedure was applied to the analysis of Dakin's solution samples, yielding results in agreement with those obtained by iodometric titration at the 95% confidence level. Copyright © 2008 John Wiley & Sons, Ltd.     

Tissue distribution and residue depletion of thiamphenicol after multiple oral dosing in seabass (Dicentrarchus labrax L.) and seabream (Sparus aurata L.)

The distribution and residue depletion of thiamphenicol (TAP) were studied in seabass ( Dicentrarchus labrax L.) and seabream ( Sparus aurata L.) reared in field conditions at temperatures of 20–28 °C. The drug was administered orally as medicated feed at the rate of 40 mg active ingredient (a.i.) kg⁻¹ of biomass once a day for 5 days. Samples of muscle, liver, skin and vertebrae from 10 fishes were collected on the 2nd and 4th day of treatment and 1, 2, 3, 5, 7, 9, 12, 15, 18, 21, 24, 27 and 30 days after the last administration of the drug, and were stored at −20 °C. Quantitative analysis of TAP was performed by high performance liquid chromatography (HPLC) after liquid-liquid extraction; the quantification limits of the HPLC method were 0.02 μg/g for muscle, and 0.05–0.10 μg/g for the other tissues. In seabass, TAP concentrations during treatment were higher in liver and muscle than in skin and vertebrae, and rapidly decreased after the end of medication. Three days after treatment ceased, TAP was still detectable in liver (0.41 ± 0.23 μg/g), vertebrae (0.09 ± 0.03 μg/g) and in three out of 10 samples of muscle (0.03 μg/g), but not in skin. All tissues were below the limits of quantification on the 5th day of withdrawal. In seabream the highest TAP concentrations during treatment were measured in liver and skin, and their reduction after the end of medication was as rapid as that of seabass: on the 3rd day after treatment ceased traces were found in only four out of 10 samples of muscle (0.03 ± 0.00 μg/g) and vertebrae (0.08 ± 0.02 μg/g).     

Release-enhancing pre-synaptic muscarinic and nicotinic receptors co-exist and interact on dopaminergic nerve endings of rat nucleus accumbens

Dopaminergic nerve endings in the corpus striatum possess nicotinic (nAChRs) and muscarinic cholinergic receptors (mAChRs) mediating release of dopamine (DA). Whether nAChRs and mAChRs co-exist and interact on the same nerve endings is unknown. We here investigate on these possibilities using rat nucleus accumbens synaptosomes pre-labeled with [³H]DA and exposed in superfusion to cholinergic receptor ligands. The mixed nAChR–mAChR agonists acetylcholine (ACh) and carbachol provoked [³H]DA release partially sensitive to the mAChR antagonist atropine but totally blocked by the nAChR antagonist mecamylamine. Addition of the mAChR agonist oxotremorine at the minimally effective concentration of 30 μmol/L, together with 3, 10, or 100 μmol/L (−)nicotine provoked synergistic effect on [³H]DA overflow. The [³H]DA overflow elicited by 100 μmol/L (−)nicotine plus 30 μmol/L oxotremorine was reduced by atropine down to the release produced by (−)nicotine alone and it was abolished by mecamylamine. The ryanodine receptor blockers dantrolene or 8-bromo-cADP-ribose, but not the inositol 1,4,5-trisphosphate receptor blocker xestospongin C inhibited the (−)nicotine/oxotremorine evoked [³H]DA overflow similarly to atropine. This overflow was partly sensitive to 100 nmol/L methyllycaconitine which did not prevent the synergistic effect of (−)nicotine/oxotremorine. Similarly to (−)nicotine, the selective α4β2 nAChR agonist RJR2403 exhibited synergism when added together with oxotremorine. To conclude, in rat nucleus accumbens, α4β2 nAChRs exert a permissive role on the releasing function of reportedly M₅ mAChRs co-existing on the same dopaminergic nerve endings.     

Nickel and rubidium uptake by whole oat plants in solution culture

Nickel and rubidium uptake by oat plants ( Avena sativa L. cv. Victory) were examined in relation to solution temperature, solution concentrations, metabolic inhibitors, anaerobic root conditions, transpiration and time. Over a 4-h period, uptake rates for both Ni²⁺ and Rb⁺ remained constant at 23°C. Decreasing temperatures to 2°C, 20 μ M concentrations of 2,4-dinitrophenol (DNP), or anaerobic root conditions decreased Ni²⁺ and Rb⁺ uptake rates by 97 to 86% in whole plants. Treatment of excised roots with 20 μ M DNP decreased Ni²⁺ uptake by 93%. Nickel and Rb⁺ uptake rates measured as a function of the external solution concentration followed a typical parabolic curve. K_m (0.012 m M ) and V_max [2.72 μmol (g dry weight)^-1 h^-1] values for Ni²⁺ were nearly 7 times lower than those for Rb⁺ [0.09 m M and 19.2 μmol (g dry weight)^-1 h^-1]. In all experiments, Ni²⁺ and Rb⁺ showed qualitatively similar uptake patterns, but Rb⁺ uptake was quantitatively more sensitive than Ni²⁺ to experimental manipulations.     

Over-reduction of cultured tobacco cells mediates changes in respiratory activities

Regulation of respiration in Nicotiana tabacum suspension cultures was studied using the respiratory inhibitor myxothiazol and the oxidative phosphorylation uncoupler carbonylcyanide p -(trifluoromethoxy)phenylhydrazone (FCCP). Myxothiazol (2 μ M ) or FCCP (2 μ M ) almost completely inhibited cell growth for about 24 h, after which the cultures could resume a growth rate similar to that of the untreated culture. During the first 18 h of myxothiazol treatment, rates of cellular respiration were similar to control cultures. The capacity of the alternative pathway was significantly higher than control 2–3 h after myxothiazol treatment and increased further after 18 h. The capacity of the alternative pathway in FCCP-treated tobacco cells also increased but to a lesser extent. ATP/ADP ratios decreased for at least 9 h after either myxothiazol or FCCP treatments, which triggered an increase in glycolytic flux and an over-reduction of cultured tobacco cells, as demonstrated by increases in levels of reactive oxygen species and cellular ethanol. The generation of reactive oxygen species served as a signal for an induction of alternative oxidase, which in turn relieved the aerobic fermentation and over-reduction of the cells.