Thermoregulation,gas exchange,and ventilation in Adelie penguins (Pygoscelis adeliae)

Summary Adelie penguins (Pygoscelis adeliae) experience a wide range of ambient temperatures (T _a) in their natural habitat. We examined body temperature (T _b), oxygen consumption ( ), carbon dioxide production ( ), evaporative water loss ( ), and ventilation atT _a from –20 to 30 °C. Body temperature did not change significantly between –20 and 20°C (meanT _b=39.3°C).T _b increased slightly to 40.1 °C atT _a=30°C. Both and were constant and minimal atT _a between –10 and 20°C, with only minor increases at –20 and 30°C. The minimal of adult penguins (mean mass 4.007 kg) was 0.0112 ml/[g·min], equivalent to a metabolic heat production (MHP) of 14.9 Watt. The respiratory exchange ratio was approximately 0.7 at allT _a. Values of were low at lowT _a, but increased to 0.21 g/min at 30°C, equivalent to 0.3% of body mass/h. Dry conductance increased 3.5-fold between –20 and 30°C. Evaporative heat loss (EHL) comprised about 5% of MHP at lowT _a, rising to 47% of MHP atT _a=30°C. The means of ventilation parameters (tidal volume [VT], respiration frequency [f], minute volume [I], and oxygen extraction [ ]) were fairly stable between –20 and 10°C (VT did not change significantly over the entireT _a range). However, there was considerable inter- and intra-individual variation in ventilation patterns. AtT _a=20–30°C,f increased 7-fold over the minimal value of 7.6 breaths/min, and I showed a similar change. fell from 28–35% at lowT _a to 6% atT _a=30°C.Abbreviations C thermal conductance - EHL evaporative heat loss - oxygen extraction - f respiratory frequency - MHP metabolic heat production - evaporative water loss - LCT lower critical temperature - RE respiratory exchange ratio - T _a ambient temperature - T _b body temperature - rate of oxygen consumption - rate of carbon dioxide production - I inspiratory minute volume - VT tidal volume     

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     

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)     

Effects of ambient temperature and altitude on ventilation and gas exchange in deer mice (Peromyscus maniculatus)

Summary The effects of different ambient temperatures (T _a) on gas exchange and ventilation in deer mice (Peromyscus maniculatus) were determined after acclimation to low and high altitude (340 and 3,800 m).At both low and high altitude, oxygen consumption ( ) decreased with increasingT _a atT _a from –10 to 30 °C. The was 15–20% smaller at high altitude than at low altitude atT _a below 30 °C.Increased atT _a below thermoneutrality was supported by increased minute volume ( ) at both low and high altitude. At mostT _a, the change in was primarily a function of changing respiration frequency (f); relatively little change occurred in tidal volume (V _T) or oxygen extraction efficiency (O₂EE). AtT _a=0 °C and below at high altitude, was constant due to decliningV _T and O₂EE increased in order to maintain high .At high altitude, (BTP) was 30–40% higher at a givenT _a than at low altitude, except atT _a below 10 °C. The increased at high altitude was due primarily to a proportional increase inf, which attained mean values of 450–500 breaths/min atT _a below 0 °C. The (STP) was equivalent at high and low altitude atT _a of 10 °C and above. At lowerT _a, (STPD) was larger at low altitude.At both altitudes, respiratory heat loss was a small fraction (<10%) of metabolic heat production, except at highT _a (20–30 °C).Abbreviations EHL evaporative heat loss - f respiration frequency - HL _a heat loss from warming tidal air - HL _e evaporative heat loss in tidal air - HL total respiratory heat loss - MHP metabolic heat production - O ₂ EE oxygen extraction efficiency - RQ respiratory quotient - T _a ambient temperature - T _b body temperatureT _lc lower critical temperature - carbon dioxide production - evaporative water loss - oxygen consumption - minute volume - V _T tidal volume     

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     

The effect of ambient temperature on the relationship between ventilation and metabolism in a small parrot (Agapornis roseicollis)

Summary Values for basal metabolism, standard tidal volume (V _T), standard minute volume ( ), and mean extraction efficiency (EO₂) in the thermal neutral zone (TNZ) inAgapornis roseicollis (1.84 ml·min^–1; 0.95 ml·br^–1, STPD; and 33.3 ml·min^–1, STPD; and 22.5%; respectively) were all very similar to values for these parameters previously measured inBolborhynchus lineola, a similarly sized, closely related species from a distinctly different habitat.Having both a lower critical temperature (T_lc) below and an upper critical temperature (T_uc) above those ofB. lineola, the TNZ ofA. roseicollis extended from 25° to at least 35°C. The thermal conductance below the TNZ ofA. roseicollis was 14% less than that ofB. lineola. Therefore, at 5°C the standard metabolic rate (SMR) of the former is 17% less than that of the latter, and at 35°C it is 20% less. At 5°CA. roseicollis has a lower EO₂ and at 35°C a higher EO₂ than that ofB. lineola. The patterns of resting energy metabolism and of ventilation ofA. roseicollis and ofB. lineola are consistent with the former species being better suited to living in a more variable thermal environment than the latter.MeanV _T has a weak positive correlation with the rate of oxygen consumption ( ) at a constant ambient temperature (T _a) but a much stronger correlation when resting increases in response to a decrease inT _a.V _t is the only ventilatory parameter which is linearly correlated toT _a from 35° to –25°C. The data suggest thatT _a may have a regulatory effect onV _T somewhat independent of or .     

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.     

Effect of oxygen on the growth (yield) of Chlorella vulgaris

The growth yield of Chlorella vulgaris, Y _kJ defined as g cells harvested per kJ of light energy absorbed by the cells, was assessed in a turbidostat culture by varying CO₂ and O₂ partial pressures ( and ). The value of Y _kJ ranged from 3.1×10^-3 to 5.0×10^-3 g cells/kJ under light-limited conditions [ = 1.02.4%, = 065%; total pressure of gas (composed of CO₂, O₂ and N₂)=1 atm]. In the light-limited environment, the algal specific growth rate deteriorated appreciably with the increase of . The deterioration accounts for the above range of Y _kJ observed. The growth inhibition due to oxygen that was defined by subtracting from 1.0 the ratio of at given values of to that at = 0% extended from 0.07–0.30 (7–30%). However, glycolate could not be detected in the turbidostat culture. Isotopic experiments on the specific rate of ¹⁴CO₂ uptake also revealed that the inhibition due to oxygen was from 22–38% when was varied from 0 to nearly 100%. These effects of oxygen were discussed, referring to the activity of ribulose-1,5-bisphosphate carboxylase that is inhibited competitively by oxygen.Non-Standard Abbreviations INH isonicotinic acid hydrazide - PPO 2,5-diphenyloxazole - DCMU 3-(-3,4-dichlorophenyl)-1,1-dimetylurea - CA carbonic anhydrase - RuP₂ ribulose-1,5-bisphosphate     

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.     

Times to exhaustion at 100% of velocity at [(V)\dot]\textO\text2 \dot V{\text{O}}_{\text{2}} max and modelling of the time-limit / velocity relationship in elite long-distance runners

The aim of this study was to measure running times to exhaustion (Tlim) on a treadmill at 100% of the minimum velocity which elicits _{max max} in 38 elite male long - distance runners _max = 71.4 ± 5.5 ml.kg^–1.min^–1 and _max = 21.8 ± 1.2 km.h^–1). The lactate threshold (LT) was defined as a starting point of accelerated lactate accumulation around 4 mM and was expressed in _max. Tlim value was negatively correlated with _max (r = -0.362, p< 0.05) and _max (r = –0.347, p< 0.05) but positively with LT (%v _max) (r = 0.378, p < 0.05). These data demonstrate that running time to exhaustion at _max in a homogeneous group of elite male long-distance runners was inversely related to _max and experimentally illustrates the model of Monod and Scherrer regarding the time limit-velocity relationship adapted from local exercise for running by Hughson et al. (1984) .     

Nitrate reductase in needles,roots and trunk wood of spruce trees [Picea abies (L.) Karst.]

Summary Nitrate reductase (EC 1.6.6.2) activity (NRA), as measured by an in vivo assay, is present in needle leaves and mycorrhizal fine root tips of adult Norway spruce [Picea abies (L.) Karst.] in at least equal amounts on a fresh weight basis, in both adult and 5-year-old trees. NRA could also be demonstrated in trunk wood of deroted trees after fertilization with 5 mM , exhibiting a longitudinal profile in the trunk. Inducibility in needles can more efficiently be achieved by NO₂ (100 g·m^-3) than by 5 mM nitrate, which is effective only in root-amputated trees. A remarkably high level of needle-NRA in unfertilized trees, which are characterized by a very low level of nitrate in the xylem sap, suggests that NRA in spruce needles may in part be constitutive. Organic-N is a major nitrogen source for the needles even in root-amputated trees, indicating pronounced exchange processes between ray parenchyma and trunk xylem, which in turn are modified by the nitrogen source fed to the trunk stump. Intact trees exhibit a very similar amino acid composition of the xylem sap, regardless of whether or has been fed. The amino acid pattern of the needles is not thrown out of balance by flooding with and , which occurs in fertilized derooted trees. This indicates a distinct potential for homoeostasis of nitrogen entrance-metabolism (i.e. NRA and glutamine synthetase activity) in the needles. In the ectomycorrhiza/fine root-system (EMC), marked differences in NRA were observed depending on root-tip diameter and along the longitudinal profile of the fine roots. EMC-nitrate reductase is strongly enhanced by . Needle-NRA exhibits a circannual rhythm. An early summer maximum is followed by a December minimum. This activity pattern matches well the transitory increase of soluble nitrogen in spring and the total protein maximum in winter. In an indirect way assimilatory NRA may well contribute to nitrogen overfertilization (by consumption of NO_X) as one possible cause of the contemporary decline of spruce populations.     

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.     

Changes in skeletal muscle oxygenation during incremental exercise measured with near infrared spectroscopy

To determine the change in muscle oxygenation in response to progressively increasing work rate exercise, muscle oxyhemoglobin + oxymyoglobin saturation was measured transcutaneously with near infrared spectroscopy in the vastus lateralis muscle during cycle ergometry. Studies were done in 11 subjects while gas exchange was measured breath-by-breath. As work rate was increased, tissue oxygenation initially either remained constant near resting levels or, more usually, decreased. Near the work rate and metabolic rate where significant lactic acidosis was detected by excess CO₂ production (lactic acidosis threshold, LAT), muscle oxygenation decreased more steeply. As maximum oxygen uptake ( ) was approached, the rate of desaturation slowed. In 8 of the 11 subjects, tissue O₂ saturation reached a minimum which was sustained for 1–3 min before was reached. The LAT correlated with both the (r = 0.95,P < 0.0001) and the work rate (r = 0.94,P < 0.0001) at which the rate of tissue O₂ desaturation accelerated. These results describe a consistent pattern in the rate of decrease in muscle oxygenation, slowly decreasing over the lower work rate range, decreasing more rapidly in the work rate range of the LAT and then slowing at about 80% of , approaching or reaching a minimum saturation at .     

Contributions of aerobic and anaerobic energy production during swimming in the bivalve molluscLimaria fragilis (family limidae)

Summary Oxygen consumption rates ( ) and a number of possible anaerobic end products were determined forLimaria fragilis at rest, and following active swimming. increased up to 8-fold (=4) during swimming. Swimming did not change the concentrations ofl-lactate, alanine or arginine phosphate in the single striated fast adductor muscle. Octopine, succinate andd-lactate were not detected in the adductor muscles of resting or active animals (<0.2 moles/g wet weight).It is concluded that the slow sustained swimming displayed byLimaria utilises predominantly aerobic mechanisms of ATP production.     

Water relations of buried eggs of mound building birds

Summary Eggs of the brush turkey (BT) and mallee fowl (MF) are incubated in mounds of soil and plant litter. Humidity in BT mounds is always near saturation (>99% RH), but in MF mounds it drops to lower values in summer (x=77% RH). Despite these high humidities, the eggs lose an average of 9.5% (BT) and 12.0% (MF) of their initial mass by evaporation before hatching. The rate of evaporation increases during incubation several-fold due to large changes in water vapor conductance of the shell and embryonic heat production. Values of in fully incubated eggs in mound material are about 3–6 times higher than values obtained from unincubated eggs in desiccators. This effect depends on two factors: (1) increases with ambient humidity, especially above 80% RH, possibly because the effective site of evaporation moves out along the walls of the pores in the eggshell. (2) Structural changes of the pores due to calcium absorption by the embryo directly increase . The first factor is most important in BT eggs and the second is dominant in MF eggs. Production of metabolic heat by the embryo increases the vapor pressure difference across the shell and further increases , especially in mounds of high humidity. The changes in pore structure are adaptive because they produce high conductances to respiratory gases and cause normal gas tensions within the egg at the end of development, yet is low enough in early development to prevent excessive water loss. Water not lost by evaporation or taken up by the embryo is stored and released during hatching. A small amount of mass is lost during incubation by respiratory gas exchange.Abbreviations BT brush turkey - MF mallee fowl - RH relative humidity     

A method for determining the maximal steady state of blood lactate concentration from two levels of submaximal exercise

The aim of this study was to estimate the characteristic exercise intensity _CL which produces the maximal steady state of blood lactate concentration (MLSS) from submaximal intensities of 20 min carried out on the same day and separated by 40 min. Ten fit male adults [maximal oxygen uptake _max 62 (SD 7) ml · min^–1 · kg^–1] exercisOed for two 30-min periods on a cycle ergometer at 67% (test 1.1) and 82% of _max (test 1.2) separated by 40 min. They exercised 4 days later for 30 min at 82% of _max without prior exercise (test 2). Blood lactate was collected for determination of lactic acid concentration every 5 min and heart rate and O₂ uptake were measured every 30 s. There were no significant differences at the 5th, 10th, 15th, 20th, 25th, or 30th min between , lactacidaemia, and heart rate during tests 1.2 and 2. Moreover, we compared the exercise intensities _CL which produced the MLSS obtained during tests 1.1 and 1.2 or during tests 1.1 and 2 calculated from differential values of lactic acid blood concentration ([1a^–]_b) between the 30th and the 5th min or between the 20th and the 5th min. There was no significant difference between the different values of _CL [68 (SD 9), 71 (SD 7), 73 (SD 6),71 (SD 11) % of _max (ANOVA test,P<0.05). Four subjects ran for 60 min at their _CL determined from periods performed on the same day (test 1.1 and 1.2) and the difference between the [la^–]_b at 5 min and at 20 min ( ([la^–]_b)) was computed. The [la^–]_b remained constant during exercise and ranged from 2.2 to 6.7 mmol · l^–1 [mean value equal to 3.9 (SD 1) mmol · l^–1]. These data suggest that the _CL protocol did not overestimate the exercise intensity corresponding to the maximal fractional utilization of _max at MLSS. For half of the subjects the _CL was very close to the higher stage (82% of _max where an accumulation of lactate in the blood with time was observed. It can be hypothesized that _CL was very close to the real MLSS considering the level of accuracy of [la^–]_b measurement. This study showed that exercise at only two intensities, performed at 65% and 80% of _max and separated by 40 min of complete rest, can be used to determine the intensity yielding a steady state of [la^–1]_b near the real MLSS workload value.     

Beta-adrenergic control of blood oxygen affinity in acutely hypoxia exposed rainbow trout

Summary Acute exposure of rainbow trout to hypoxic water ( =40 mmHg, 15 °C) caused a significant (P<0.01) increase in blood O₂ affinity, from the normoxicP ₅₀ value (at pH_e 7.93) of 23.2±1.1 mmHg to about 19 mmHg, within 5 min. Specimens injected with the -antagonist propranolol showed no change in bloodP ₅₀, despite a more pronounced reduction of arterial during the hypoxic exposure.The change in bloodP ₅₀ coincided with an increase in plasma catecholamines, notably noradrenaline. There was no change in the molar ratios of ATPHb₄ and GTPHb₄. The altered bloodP ₅₀, however, correlated with an alkalinization and an increased sodium concentration of the red cells. This red cell alkalinization can be explained by -adrenergic stimulation of a membrane bound Na⁺/H⁺ antiporter.Propranolol injection into normoxic resting trout caused a significant decrease in and increase in indicating -adrenergic control of gas exchange in the gills.     

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.     

rDNA magnification in D. melanogaster: state of rDNA copies following the first step.

Summary D. melanogaster males of bb/O genetic constitution undergoing rDNA magnification were mated singly to XXbb ⁺/O females, yielding bb/O male progeny, and to X_NO-w sn bb ⁺ fameles, yielding bb/X_NO- females. The male and female offspring were scored for the bb ⁺ phenotype.Results show that there is a higher percentage of bb ⁺ flies in the bb/O male progeny than in bb/X_NO- females progeny, in single crosses as well as in the combined data. rRNA/DNA hybridization experiments agree with this observation, by showing that the rDNA content in the progeny of premagnified flies was higher in the sons than in the daughters.These data indicate that the increase of ribosomal RNA genes is not due to a stable event such as an unequal mitotic sister exchange, whereas they do not contrast with the extracopy model.     

Determination of heteronuclear three-bond J-coupling constants in peptides by a simple heteronuclear relayed E.COSY experiment

Summary A simple heteronuclear relayed E.COSY pulse sequence with a minimum number of pulses is proposed for the quantitative determination of heteronuclear three-bond J-coupling constants in uniformly ¹³C-enriched polypeptide samples. Numerous heteronuclear three-bond coupling constants, including , , , and , can be determined for each residue from a single heteronuclear relayed E.COSY spectrum. Couplings relevant for stereospecific assignments as well as for the determination of dihedral angles in the amino acid backbone and in side chains are obtained. The method is demonstrated on the uniformly ¹³C-enriched decapeptide antamanide (-Val¹-Pro²-Pro³-Ala⁴-Phe⁵-Phe⁶-Pro⁷-Pro⁸-Phe⁹-Phe¹⁰-).