(H)NCAHA and (H)CANNH experiments for the determination of vicinal coupling constants related to the ϕ-torsion angle

Summary A set of three-dimensional triple-resonance experiments is described which provide , , and coupling constants. The pulse sequences generate E.COSY-like multiplet patterns and comprise a magnetization transfer from the amide proton to the α-proton or vice versa via the directly bound heteronuclei. For residues with the ¹H^α spin resonating close to the H₂O signal, a modified HNCA experiment can be employed to measure the vicinal ¹H^N,¹H^α couplings. Ambiguities associated with the conversion of values into ϕ-angle constraints for protein structure determination can be resolved with the knowledge of the heteronuclear ³J-couplings. In favourable cases, stereospecific assignments of glycine α-protons can be obtained by employing the experiments described here in combination with NOE data. The methods are applied to flavodoxin from Desulfovibrio vulgaris.     

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.     

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.     

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)     

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.     

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     

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     

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.     

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     

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) .     

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.     

Measurement of four-bond HN-Hα J-couplings in staphylococcal nuclease

Summary Quantitative J-correlation and triple-resonance ECOSY-type experiments are used to unambiguously establish the presence of four-bond sequential H^N-H J-couplings in the protein staphylococcal nuclease. Substantially negative values, ranging from -0.8 to -2.3 Hz, are observed when the angle is near +120°, and the following angle near +60°. For other conformations, the four-bond H^N-H J-couplings fall between -0.5 and +0.5 Hz.     

Prediction of individual oxygen uptake on-step transients from frequency responses

Power-oxygen uptake ( ) frequency responses can be used to predict responses to arbitrary exercise intensity patterns. It is still an open question for which range of exercise intensities such computed response patterns yield valid predictions. In the present study, we determined the power- frequency response of nine sports students by means of pseudo-randomised switching between 20 W and 80 W during upright and supine cycle exercise. Starting from a baseline of 20 W each subject also performed sustained step increases to 40 W, 80 W, 120 W, and 160 W in both positions. The individual step responses were then compared with the expected time-courses predicted on the basis of the individual frequency responses. The comparison showed a close agreement for the 20 W–40 W and 20 W–80 W steps in both positions. With larger step amplitudes the kinetics became increasingly slower than the predicted time course in both positions. During additional ramp tests (10 W · 30 s^–1) whole blood lactic acid concentration [1a^–]_b tended to be higher in the supine position at exercise intensities higher than 160 W. The mean power at 4 mmol · 1^–1 [la^–]_b amounted to 234 (SD 32) W and 253 (SD 44) W (P<5%) in the supine and the upright position, respectively. The maximal oxygen uptake relative to body mass was not found to be significantly different [upright, mean 57 (SD 10) ml · (min · kg)^–1;supine, mean 54 (SD 10) ml · (min · kg)]. These findings would suggest that for a range of mild exercise intensities kinetics are not appreciably influenced by the step amplitude or by cardiovascular changes associated with the upright and the supine position.     

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     

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.     

Are oxygen uptake kinetics at the onset of exercise speeded up by local metabolic status in active muscles?

To assess the rate-limiting factor of oxygen uptake ( ) kinetics at the onset of exercise, six healthy male sedentary subjects performed repeated one-legged constant-load cycle exercise. The one-legged constant-load exercise test consisted of two 5-min periods of pedalling at an exercise intensity of 50 W, with a 5-min rest between periods (these exercise periods, i.e. first and second exercises, were performed by the same leg). The exercise was then repeated using the other leg. In addition, two-legged incremental exercise was investigated to establish whether kinetics were affected by slower heart rate kinetics. The incremental exercise test consisted of two-legged pedalling first with the cycle unloaded as a warm-up for 5 min followed by 50-W exercise for 5 min. The exercise intensity was then increased to 100 W for 5 min. During exercise, gas exchange parameters were determined by the breath-by-breath method and cardiac output ( ) was determined continuously by an impedance technique with a computer-based automated system. To determine the kinetics of heart rate (HR), and , a best fit procedure was employed using least-squares criteria with a time delay, except during the initial increase. During the one-legged constant-load exercise test, kinetics were significantly accelerated by repeated exercise using the same leg. On the other hand, when the exercise was changed to the other leg, kinetics were significantly slower, although kinetics continued to be faster. During the incremental exercise test, although the HR response was slower at the transition from 50-W to 100-W exercise than at the transition from warm-up to 50-W exercise, there were no significant changes in kinetics. These findings suggest that kinetics may be affected by metabolic conditions in the muscle, but not by blood flow ( and/or HR) kinetics.     

Regulation of mineral nitrogen uptake in plants

In the biosphere plants are exposed to different forms of N, which comprise mineral and organic N forms in soils as well as gaseous NH3, NOx, and molecular N2 in the atmosphere. The form of N uptake is mainly determined by its abundance and accessibility, which make and the most important N forms for plant nutrition under agricultural conditions. With minor importance, the form of N uptake is also subject to plant preferences, by which plants maintain their cation/anion balance during uptake. However, some species seem to have an obligatory preference which even prevents their growth on certain other N sources. In general, uptake of a certain N form closely matches the growth-related demand of the plant, at least when N transport to the root surface is not limiting. In addition, many plants accumulate large pools of N during vegetative growth which are remobilized in the generative stage. As a consequence, systems responsible for N transport need to be tightly regulated in their expression and activity upon sensing N availability and plant demand. Employing the tools of molecular genetics, the first plant genes encoding transporters for inorganic N have recently been isolated and characterized. These data can now complete the wealth of physiological and nutritional studies on N uptake. The present article will focus on the uptake of and into root cells and tries to link data derived from physiological, genetic and molecular studies.     

Laser light scattering determination of size and dispersity of synaptosomes and synaptic vesicles isolated from squid (Loligo pealei) optic lobes

Quasi-elastic laser light scattering has been used to investigate the size and dispersity of synaptosomes and synaptic vesicles isolated from optic lobes of the squid Loligo pealei. Synaptosomal fractions were highly polydisperse ( ) and the mean diameter ( ) ranged from 0.5–2.0 m. Size distribution histograms yielded two major components — smaller particles ( ) and a larger group of particles ( ). The heterogeneity of the synaptosomal particles detected in solution is in agreement with published data obtained using electron microscopy. Purified synaptic vesicle fractions also yielded complex particle size distribution data. A component with a mean diameter in the range 150–250 nm was detected, though a smaller particle ( ) dominated the scattering signal. This smaller particle closely resembles in size the electron lucent vesicles seen in the majority of squid optic lobe nerve terminals when examined by electron microscopy. Osmotically-induced shirnkage and swelling of the synptosomes was detected. Depolarization by veratridine (1.0×10^–4 M) did not result in a detectable change in the size of synaptosomal particles.     

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.     

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.