Research into pharmacological and metabolic dependence of delayed inactivated outward potassium current at the snail neuronal somatic membrane

Features of pharmacological and metabolic sensitivity of potassium outward current dependent on the presence of Ca ions in the intracellular solution were investigated. Verapamil was found to produce depression and accelerated inactivation of this current. The blocking action of verapamil intensifies with an increase in extracellular concentration of calcium ions. Functioning of channels depends on intracellular metabolic processes. Current level declined during intracellular perfusion of the neuron with artificial saline. Reducing the temperature of the perfusing solution to 10°C produced a similar result. This decline could be avoided by introducing 2 mM ATP and 3 mM Mg ions into the intracellular solution; current decay could actually be reversed by intracellular application of exogenous protein kinase C. Polymyxin B (10^–4 M), a protein kinase C blocker, depressed the component of outward potassium current dependent on extracellular Ca²⁺. Mechanisms possibly underlying Ca²⁺ action on via a phosphatidyl inositol metabolism is discussed in the light of the experimental findings obtained.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 21, No. 1, pp. 52–59, January–February, 1989.     

Quantitative analysis of outward rectifying K+ channel currents in guard cell protoplasts fromVicia faba

Summary A quantitative analysis of the time and voltage dependence of outward-rectifying K⁺ currents ( ) in guard cells fromVicia faba is described using the whole-cell patch-clamp technique. After step depolarizations from –75 mV to potentials positive to –40 mV, time-dependent outward currents were produced, which have recently been identified as K⁺ channel currents. This K⁺ current was characterized according to its time dependence and its steady-state activation. could be described in terms of a Hodgkin-Huxley type conductance. Activation of the current in time was sigmoid and was well fitted by raising the activation variable to the second power. Deactivating tail currents were single exponentials, which suggests that only one conductance underlies this slow outward K⁺ current. Rates of channel closing were strongly dependent on the membrane potential, while rates of channel opening showed only limited voltage dependence leading to a highly asymmetric voltage dependence for channel closing and opening. The presented analysis provides a quantitative basis for the understanding of channel gating and channel functions in plant cells.     

Dynamics of cardiorespiratory function in Standardbred horses during different intensities of constant-load exercise

Summary Six Standardbred horses were used to evaluate the time course of pulmonary gas exchange, ventilation, heart rate (HR) and acid base balance during different intensities of constant-load treadmill exercise. Horses were exercised at approximately 50%, 75% and 100% maximum oxygen uptake ( max) for 5 min and measurements taken every 30 s throughout exercise. At all work rates, the minute ventilation, respiratory frequency and tidal volume reached steady state values by 60 s of exercise. At 100% max, the oxygen consumption ( ) increased to mean values of approximately 130 ml/kg·min, which represents a 40-fold increase above resting . At the low and moderate work rates, showed no significant change from 30 s to 300 s of exercise. At the high work rate, the mean at 30 s was 80% of the value at 300 s. The HR showed no significant change over time at the moderate work rate but differing responses at the low and high work rates. At the low work rate, the mean HR decreased from 188 beats/min at 30 s to 172 beats/min at 300 s exercise, whereas at the high work rate the mean HR increased from 204 beats/min at 30 s to 221 beats/min at 300 s exercise. No changes in acid base status occurred during exercise at the low work rate. At the moderate work rate, a mild metabolic acidosis occurred which was nonprogressive with time, whereas the high work rate resulted in a progressive metabolic acidosis with a base deficit of 16 mmol/l by 300 s exercise. It is concluded that the kinetics of gas exchange during exercise are more rapid in the horse than in man, despite the relatively greater change in in the horse when going from rest to high intensity exercise.Symbols and abbreviations E minute ventilation - V _T tidal volume - oxygen uptake - carbon dioxide output - oxygen pulse - ventilatory equivalent for oxygen - ventilatory equivalent for carbon dioxide - R respiratory exchange ratio - HR heart rate - SBC standard bicarbonate - STPD standard temperature and pressure dry - BTPS body temperature and pressure saturated - arterial oxygen content - arteriovenous oxygen content difference - Rf respiratory frequency     

Proportionality of ELF electric field-induced growth inhibition to induced membrane potential inZea mays andVicia faba roots

Summary The postulate that 60-Hz electric field-induced bioeffect severity is proportional to induced transmembrane potential [ ] magnitude was tested and supported using a plant root model cell system. Statistically significant correlations were obtained upon regression of the relative rates of exposedVicia faba andZea mays root segment growth on the average (calculated) arising in those segments under specified 60 Hz field exposure conditions. The associated with the apparent threshold for growth inhibition was similar inZea andVicia roots (2.5 vs 2.4 mV, respectively). At greater than this threshold,Zea root growth declined by about 9% per mV, andVicia root growth by about 19% per mV induced potential.Abbreviations EF electric field - RGR relative growth rate - RSGR relative segmental growth rate - induced membrane potential - segmental-average induced membrane potential - VC _d region of root tip in which a complete, nascent vascular cylinder is first distinguishable in histological sections     

Skeletal muscle [(V)\dot]O2 \dot V_{O_2 } in rat and lemming: Effect of blood flow rate

Summary The rate of oxygen consumption ( ) by skeletal muscle was investigated in isolated perfused hindlimbs of laboratory rats and lemmings (Lemmus). In both species, increased in proportion to blood flow rate, even at flow rates 4–5 times above resting level. The slope of the line relating to skeletal muscle blood flow was significantly greater in the lemming than in the rat. This may be related to the inverse relationship between body weight and metabolic rate. These data support the hypothesis that in small animals a dependent relationship exists between blood flow and skeletal muscle .     

Activity and oxygen consumption during hypoxic exposure in high altitude and lowland sceloporine lizards

Summary Resting rates of O₂ consumption against , exercise endurance times and during recovery from vigorous exercise were measured inSceloporus occidentalis captured near sea level and inS. graciosus captured above 2850 m. Oxygen consumption against was also measured inS. occidentalis captured above 2850 m. When was recorded continuously, as ambient was slowly reduced from 155 Torr, it became directly dependent upon ambient between 110 and 120 Torr. The critical for the high altitude lizards was lower than that for the lowland lizards, which enabled the former to maintain relatively higher 's when ambient was reduced below 120 Torr. The high altitude lizards also had significantly greater endurance when stimulated to exercise at 1600 m ( 130 Torr). Both the higher under hypoxia and the greater endurance roughly parallel a significantly greater maximum in the high altitude lizards. At a simulated altitude of 3600 m ( 100 Torr), maximum and rate of recovery of the O₂ debt calculated from post active were significantly reduced in the lowland but not the high altitude lizards. The effects of simulated altitude conditions on the lowland but not the mountaine animals indicate adaptations to altitude in these sceloporine lizards. We did not find any consistent relationship between organ/body weight ratios or hematocrit and our measures of endurance or the altitude at which the lizards were captured.     

Respiration of the emersed shore crab at variable ambient oxygenation

Summary In the intertidal shore crab,Carcinus maenas, pH and values of the prebranchial venous hemolymph, and , the PO₂ values of the arterialized cardiac hemolymph, , were measured, and the ventilatory activity was assessed by measuring the hydrostatic pressures at the exits of the epibranchial cavities, under four environmental conditions: normoxic water, normoxic air, hypoxic gas, hyperoxic gas.In the crab breathing normoxic air, was lower and higher than in animals breathing normoxic water. With the switch to hypoxic gas, ventilation increased and and decreased. With the switch to hyperoxic gas, ventilation decreased, and increased and decreased.Thus these crabs breathing air were not as well oxygenated as when breathing air-equilibrated water, and because of their low , they were relatively hypervetilating and hypocapnic compared to air-breathing vertebrates. Hyperoxic breathing, increasing and reducing ventilatory drive, led to increased . Conversely, was reduced by hypoxic breathing. These observations suggest that the gas exchanger of intertidal crabs is not as successfully designed for air breathing as that of land-colonizing insects and air-breathing vertebrates.     

Gill function in an elasmobranch

Summary Highly efficient oxygen uptake in elasmobranchs, as indicated by frequent excess of over has previously been ascribed to the operation of multicapillary rather than counter-current gas exchange by the gills. Analysis of models shows that, at maximum efficiency, a multicapillary system cannot account for values of greater than . In Port Jackson sharks Heterodontus portusjacksoni) commonly exceeds , which indicates the operation of a functional counter-current at the respiratory surface. The anatomical basis of this counter-current is provided by the demonstration that a continuous flow of water passes between the secondary lamellae into septal canals and thence via the parabranchial cavities to the exterior.Queen Elizabeth II Fellow.     

Breathing pattern and growth: comparative aspects

Summary The resting oxygen consumption and breathing pattern of nine newborn and adult species (ranging in body size from mouse to human) have been compared on the basis of data collected from the literature. Minute ventilation is similarly linked to at both ages, the percent of extracted as O₂ about 2.2. Tidal volume/kg is an interspecies constant in newborns and adults, approximately 8 ml/kg. Breathing frequency decreases with the increase in size in a different way at the two ages: large species have newborns breathing at rates 2–3 times above the corresponding adults' values, while in the small species newborns and adults breathe at almost the same rate. Therefore the newborns of the smallest species have both and below the expected values, implying a greater inability to cope with the external demands than newborns of larger species. Several considerations indicate that in the smallest newborns the mechanical properties of the respiratory system could be a constraint to resting ventilations larger than observed. It is therefore possible that their low is the cause, and not the effect, of the relatively small .     

Estimation of surface charges in some biological membranes

Summary The resting membrane potential data existing in the literature for the giant axon of the squid, frog muscle and barnacle muscle have been analyzed from the standpoint of the theory of membrane potential due to Kobatake and co-workers. The average values derived for the effective charge density (where is a constant, , and represents the fraction of counterions that are free, and is the stoichiometric charge density in the membrane) present on the different biomembranes existing in their normal ionic environment are 0.3, 0.325 and 0.17 M for the squid axon, frog and barnacle muscles, respectively. On the assumption that the values of are 0.4 and 0.2 for nerve and muscle membranes, respectively, values of 0.75, 1.62 and 0.85 M have been derived for the stoichiometric charge density present in the respective biological membranes. These correspond to 1 negative charge per 222, 103 and 195 Å of the membrane area of the squid axon, frog and barnacle muscles, respectively.     

Aerobic metabolism of the lizardVaranus exanthematicus: Effects of activity, temperature, and size

Summary Oxygen consumption was measured at rest and during spontaneous activity at body temperatures of 25 and 35°C in 14 fasting Savanna monitor lizards,Varanus exanthematicus ranging in weight from 172 to 7500 g. The allometric relationship between metabolic rate at 25°C and body weight (W) is given by: (ml O₂ STPD·g^–1·hr^–1)=0.88W ^–0.43 (Fig. 2). Although statistical comparisons are equivocal, this intraspecific size dependence exceeds that reported for interspecific comparisons among reptiles and other vertebrate groups (Fig 3). A reproducible diurnal pattern of activity was observed in undisturbed animals with minimum values of between 2400 and 0800 h (Fig. 1). Spontaneous activity and generally reached peak values between 1200 and 2000 hrs. The average ratio of active aerobic metabolic rate (AMR) to minimum (standard) aerobic metabolic rate (SMR) was 8.2. This voluntary AMR/SMR inVaranus exceeds the AMR/SMR for most reptiles stimulated to exhaustion. The high aerobic capacity is consistent with other evidence for efficient exchange and transport of respiratory gases inV. exanthematicus; e.g., low or absent intracardiac shunt flow resulting in high arterial saturation and low ventilation and perfusion requirements.     

Control of acid-base status in active and dormant land snails,Otala lactea (Pulmonata,Helicidae)

Summary Control of extracellular acid-base status was examined during activity and dormancy inOtala lactea (Pulmonata, Helicidae). Active snails showed little variation in hemolymph pH and at constant temperature. With increase of temperature, hemolymph increased from about 6 Torr at 5°C to 13 Torr at 24°C and pH decreased by about 0.017 pH units/°C, a pattern consistent with alphastat regulation of pH via ventilatory control of .During dormancy, mean hemolymph increased to about 50 Torr. Venous pH declined by about 0.4 units due to hypercapnia and fluctuated more widely than in active snails due to variability of . Hemolymph pH declined further in prolonged dormancy due to progressive metabolic acidosis; after one year of dormancy the mean hemolymph pH was about 0.8 units lower than that of active snails at similar temperature.Active snails exposed experimentally to high showed a large increase in hemolymph [HCO ₃ ^– ]. However, [HCO ₃ ^– ] declined by up to 50% during dormancy, despite the naturally occurring hypercapnia. Hemolymph osmolality and the concentrations of solutes other than [HCO ₃ ^– ] increased with increasing duration of dormancy. Concentrations of magnesium and calcium increased about 2.5 times more rapidly than those of sodium and chloride, indicating that acidosis is partially offset by the dissolution of carbonates from the shell or tissues.     

Changes in Swallowing-Related Tachycardia and Respiratory Arrhythmia Induced by Modulation of Tonic Parasympathetic Influences

In 19 tested persons in the resting lying position, we examined changes in the mean duration of R-R intervals and in parasympathetic chronotropic cardiac reactions (respiratory arrhythmia,RA, and swallowing-related tachycardia, ST) induced by intensified tonic parasympathetic influences after peroral administration of antagonists of muscarinic (M) receptors in small doses. Administration of 0.02 g of an extract of Belladonna, which contains alkaloids of the atropine group, led to a significant increase (P < 0.001). In this case, the RA at the natural frequency of breathing (P < 0.05) and ST (P < 0.001) simultaneously increased. Close correlation was observed both between initial values of the ST and in various tested persons (r = 0.87) and between changes in these values, which were induced by the action of small amounts of M antagonists (r = 84). In various persons, the RA during deep slowed breathing (6 min^-1, RA₆) demonstrated ambiguous modifications. In the structure of RA₆, we identified two components: an inspiratory tachycardic (TC) and an expiratory bradycardic (BC). With intensification of the parasympathetic influences, the TC began to prevail; its increase was strongly related to a rise in the (r = 0.84). In contrast, correlation between changes in the BC and an increase in the was completely absent (r = 0.001). Therefore, small amounts of M antagonists intensify both the tonic component of parasympathetic control (increasing the ) and its dynamic indices, the magnitudes of the ST and RA. Tachycardia during swallowing and the inspiratory TC under conditions of deep slowed breathing are closely related to changes in the parasympathetic influences. The magnitude of the RA is the index, which reflects the tonic component of chtonotropic control under conditions of both slowed and natural breathing less adequately than the ST.     

The relationship between the energetic efficiency in different micro-organisms and the rate and type of metabolite overproduced

Summary Data regarding the degree of energy conservation as determined by the and the highest rates of metabolite production reported for various micro-organisms have been collated and analysed. The results have indicated that the highest rates of metabolite production occur in micro-organisms possessing low efficiencies of energy conservation. Moreover, in the case of exopolysaccharide production the oxidation state of the polymer is inversely related to the value of the producing organism. In general, the rate of ATP turnover associated with exopolysaccharide production or the potential rate associated with over-production of other metabolites is inversely related to the value of the producing organism. Analysis of current production rates for a range of metabolites suggests that there is scope for major improvements of existing processes by careful selection of appropriate micro-organisms.     

Water economy in bird nests

Summary Nest humidity ( ) under an incubating bantam hen was measured at ambient ranging from 1.3 to 25.0 mmHg. Weight loss of eggs was recorded in clutches of varying size. Nest and ambient were also measured in wild incubating willow ptarmigan nests in dry and wet habitats.Nest increased linearly with ambient in a way predictable on the assumption that the water vapour conductance ( ) of brood patch skin, plumage and eggs were constant and independent of ambient . Nest was also dependent of clutch size. Egg dehydration was quantitatively predicted from measured values and the laws of diffusion.Our findings confirm earlier conclusions that the adult bird does not actively regulate nest at varying ambient . Birds can presumably achieve appropriate egg dehydration by a strategy combining nest site, nest construction, egg shell conductance and incubation behaviour which meets the requirements of their breeding climate.Abbreviations water vapur pressure - water vapur conductance - water flux     

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     

Growth and product formation of actinomycetes cultivated at increased total pressure and oxygen partial pressure

Summary Influence of pressure (P) and oxygen partial pressure ( ) on cultivation of various Streptomyces spp. and Micromonospora purpurea was examined in pressurized air-lift and stirred tank fermenters. The maximum was 2100 mbar. Growth and product formation of all cultures tested were markedly influenced by higher than 1000 mbar. There is evidence that wild strains are more oxygen tolerant than production strains. At a certain the metabolic activities of all cultures were inhibited. However, results obtained with S. aureofaciens and S. rimosus indicated an increase in specific product formation rate at elevated pressure. With increase in oxygen tension incorporation of oxygen into tetracycline molecules was enhanced. Since elevated oxygen tension can either show inhibiting effects or may be used for regulation of product formation and selectivity, the influence of should be determined in an appropriate experimental set-up for each process.Offprint requests to: U. Onken     

Gas exchange in the incubation mounds of megapode birds

Summary The brush turkey (Alectura lathami) and mallee fowl (Leipoa ocellata) are megapode birds that incubate their eggs by burying them in mounds. Respiratory gas exchange between the buried eggs and the atmosphere occurs mainly by diffusion through about 60 cm of decomposing forest litter (brush turkey) or sand (mallee fowl).Gas fluxes in the brush turkey mound are greatly influenced by the respiration of thermophilic microorganisms which consume O₂ at rates over eight times that of all of the eggs. The respiratory exchange ratio ( ) of the microorganisms is 0.75 and theQ ₁₀ for metabolism is 2.56. Fermentation and nitrogen fixation do not occur in the mounds.If the mound becomes too wet, gas tensions near the eggs can become critical because water increases rates of microbial respiration and impedes gas diffusion. However, field mounds are relatively dry, possibly because the adult bird modifies the shape of the mound and affects the entry of rain water. At egg level in field mounds, and are about 132 and 21 Torr, respectively, in both species. Embryonic respiration decreases and increases about 5 Torr in the immediate environment of individual eggs in late development. Due to a high eggshell gas conductance, which increases during incubation, the gas tensions within the shell of late embryos ( ca. 108 Torr, ca. 47 Torr) are not far from the mean values found in species that nest above ground.     

Altitudinal and seasonal effects on aerobic metabolism of deer mice

Summary I compared the maximal aerobic metabolic rates ( ), field metabolic rates (FMR), aerobic reserves ( -FMR), and basal metabolic rates (BMR) of wild and recently captured deer mice from low (440 m) and high (3800 m) altitudes. To separate the effects of the thermal environment from other altitudinal effects, I examined mice from different altitudes, but similar thermal environments (i.e., summer mice from high altitude and winter mice from low altitude). When the thermal environment was similar, , FMR, and aerobic reserve of low and high altitude mice did not differ, but BMR was significantly higher at high altitude. Thus, in the absence of thermal differences, altitude had only minor effects on the aerobic metabolism of wild or recently captured deer mice.At low altitude, there was significant seasonal variation in , FMR, and aerobic reserve, but not BMR. BMR was correlated with , but not with FMR. The significant positive correlation of BMR with indicates a cost of high , because higher BMR increases food requirements and energy use during periods of thermoneutral conditions.Abbreviations BMR basal metabolic rate - FMR field metabolic rate - partial pressure of oxygen - T _a ambient temperature - T _b body temperature - T _e operative temperature - maximal aerobic metabolic rate     

Energy requirements of Adélie penguin (Pygoscelis adeliae) chicks

Summary The energy requirements of Adélie penguin (Pygoscelis adeliae) chicks were analysed with respect to body mass (W, 0.145–3.35 kg, n=36) and various forms of activity (lying, standing, minor activity, locomotion, walking on a treadmill). Direct respirometry was used to measure O₂ consumption ( ) and CO₂ production. Heart rate (HR, bpm) was recorded from the ECG obtained by both externally attached electrodes and implantable HR-transmitters. The parameters measured were not affected by hand-rearing of the chicks or by implanting transmitters. HR measured in the laboratory and in the field were comparable. Oxygen uptake ranged from in lying chicks to at maximal activity, RQ=0.76. Metabolic rate in small wild chicks (0.14–0.38 kg) was not affected by time of day, nor was their feeding frequency in the colony (Dec 20–21). Regressions of HR on were highly significant (p< 0.0001) in transmitter implanted chicks (n=4), and two relationships are proposed for the pooled data, one for minor activities ( ), and one for walking ( ). Oxygen consumption, mass of the chick (2–3 kg), and duration of walking (T, s) were related as , whereas mass-specific O₂ consumption was related to walking speed (S, m·s^-1) as .Abbreviations bpm beats per minute - D distance walked (m) - ECG electrocardiogram - HR heart rate (bpm) - ns number of steps - RQ respiratory quotient - S walking speed (m·s^-1) - T time walked (s) - W body mass (kg)