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Abstract Background Crystallography is a data-rich, software-intensive scientific discipline with a community that has undertaken direct responsibility for publishing its own scientific journals. That community has worked actively to develop information exchange standards allowing readers of structure reports to access directly, and interact with, the scientific content of the articles. Results Structure reports submitted to some journals of the International Union of Crystallography (IUCr) can be automatically validated and published through an efficient and cost-effective workflow. Readers can view and interact with the structures in three-dimensional visualization applications, and can access the experimental data should they wish to perform their own independent structure solution and refinement. The journals also layer on top of this facility a number of automated annotations and interpretations to add further scientific value. Conclusions The benefits of semantically rich information exchange standards have revolutionised the scholarly publishing process for crystallography, and establish a model relevant to many other physical science disciplines.

Decapod crustaceans inhabit aquatic environments that are frequently subjected to changes in salinity and oxygen content. The physiological responses of decapod crustaceans to either salinity or hypoxia are well documented; however, there are many fewer reports on the physiological responses during exposure to these parameters in combination. We investigated the effects of simultaneous and sequential combinations of low salinity and hypoxia on the cardiovascular physiology of the Dungeness crab, Cancer magister. Heart rate, as well as haemolymph flow rates through the anterolateral, hepatic, sternal and posterior arteries were measured using a pulsed-Doppler flowmeter. Summation of flows allowed calculation of cardiac output and division of this by heart rate yielded stroke volume. When hypoxia and low salinity were encountered simultaneously, the observed changes in cardiac properties tended to be a mix of both factors. Hypoxia caused a bradycardia, whereas exposure to low salinity was associated with a tachycardia. However, the hypoxic conditions had the dominant effect on heart rate. Although hypoxia caused an increase in stroke volume of the heart, the low salinity had a more pronounced effect, causing an overall decrease in stroke volume. The patterns of haemolymph flow through the arterial system also varied when hypoxia and low salinity were offered together. The resulting responses were a mix of those resulting from exposure to either parameter alone. When low salinity and hypoxia were offered sequentially, the parameter experienced first tended to have the dominant effect on cardiac function and haemolymph flows. Low salinity exposure was associated with an increase in heart rate, a decrease in stroke volume and cardiac output, and a concomitant decrease in haemolymph flow rates. Subsequent exposure to hypoxic conditions caused a slight decrease in rate, but other cardiovascular variables were largely unaffected. In contrast, when low salinity followed acclimation to hypoxic conditions, apart from an increased heart rate, there were no other cardiovascular changes associated with the low salinity episode. The implications of these changes in cardiovascular dynamics are discussed in relation to physiological mechanisms and the ecology of decapod crustaceans, in hypoxic or low salinity environments., (Copyright 2003, Wiley-Liss, Inc.)

The larval development of penaeid shrimp is among the most complicated in crustaceans. In Metapenaeus ensis, there are six naupliar, three protozoeal and three mysid larval instars, followed by postlarval development. Irregular heartbeat begins late in naupliar instar 6. Co-ordinated beating at 400-600 beats min(-1) commences in the first protozoeal instar and continues throughout larval life. Initially, the contractile region is located more posteriorly in the cephalothorax and has a single pair of ostia, and the arterial distribution is limited to a single anterior vessel. In later mysid instars, a second cardiac pumping site develops posterior to, but connected with, the original site. This extension is more muscular, contains additional ostia and develops additional distribution vessels supplying the cephalothorax and abdominal areas. The original site is gradually merged into the new extension and only small refinements in the circulation occur in postlarval and juvenile life. Changes in physiological responses of the heart also occur throughout development. Responses to intra-pericardial microinjection of 5-hydroxytryptamine change drastically during development, as do cardiac responses to ambient hypoxia. Similarly, heartbeat of later juvenile instars is inhibited by injection of tetrodotoxin, while heartbeat of larval and early juvenile instars is not, suggesting that neurogenic regulation via the cardiac ganglion arises later in development. Our present studies attempt to integrate the anatomical and physiological changes in the development of the crustacean heart.

Crustaceans are often tolerant of hypoxic exposure and many regulate O(2) consumption at low ambient O(2). In acute hypoxia, most increase branchial water flow, and many also increase branchial haemolymph flow, both by an increase in cardiac output and by shunting flow away from the viscera. The O(2)-binding affinity of crustacean O(2) carriers increases in hypoxic conditions, as a result of hyperventilation induced alkalosis. In chronic hypoxic exposure some crustaceans do not sustain high ventilatory pumping levels but increased effectiveness of O(2)-uptake across the gills is maintained as a result of the build up of metabolites such as lactate and urate which also function to increase the haemocyanin O(2)-binding affinity. Chronic exposure to hypoxia also may increase O(2)-binding capacity and promote the synthesis of new high O(2)-affinity carrier molecules. Exposure to untenable rates or levels of O(2) depletion causes many decapodan crustaceans to surface and ventilate the gills with air. Burrowing crayfish provide an example of animals, which excel in all these mechanisms. Control mechanisms involved in compensatory responses to hypoxia are discussed.

Work in the last decade has shown that crustacean open circulatory systems are highly efficient and controlled in a complex manner. Control occurs at several levels. Myocardial contraction is initiated in the cardiac ganglion but constantly modulated by the central nervous system, both directly via the cardioregulatory nerves and indirectly via the neurohormonal system. Heart rate and stroke volume can be controlled independently and measurements of both are needed to assess cardiac output accurately. Haemolymph outflow from many arthropod hearts is via a complex multiarterial distribution system, and the regional distribution of cardiac output is tightly controlled via cardioarterial valves at the base of each artery. These valves contain innervated muscle, and differential contraction serves to regulate the efflux of oxygenated haemolymph into a particular system. The major influence on both the evolution and control of arthropod open blood vascular systems is efficiency of oxygen uptake and delivery. This influence is illustrated by reference to a variety of crustacean and other arthropod types.

Peptide neurohormones exist as functionally similar analogues in a wide variety of invertebrate and vertebrate phyla, and many have been implicated as cardiovascular regulators. In decapod crustaceans, these include the pentapeptide proctolin, crustacean cardioactive peptide (CCAP) and the FMRF amide-related peptides F1 and F2, all of which are found in the pericardial organs located immediately upstream of the heart. Cardioexcitatory activity has been demonstrated by these four peptides in both isolated and semi-isolated arthropod hearts; CCAP, however, has minimal effects on the heart of Cancer magister. In the present study, we determined the effects of proctolin, F1 and F2 on the heart of the crab C. magister in both in vitro (semi-isolated heart) and in vivo (whole animal) preparations. In semi-isolated hearts, infusion of each peptide caused cardioexcitation, increasing the rate and stroke volume of the heart. In whole crabs, the peptides were cardioinhibitory; the strongest effects were observed with F1 and F2, which dramatically decreased heart rate, cardiac stroke volume and cardiac output. These results cast doubt on current perceptions of the functional role of cardioactive peptides in the regulation of invertebrate cardiovascular performance in vivo.

The FMRFamide-related peptides F1 and F2, originally isolated from lobster pericardial organs, have been shown to exert cardioexcitatory effects on isolated or semi-isolated crustacean hearts. The present study sought to determine the in vivo effects of F1 and F2 on cardiac and circulatory performance of Cancer magister using a pulsed-Doppler technique. In general the effects of F1 and F2 were similar; however, F1 was more potent and its effects were of longer duration than those exerted by F2. Infusion of either F1 or F2 caused a decrease in heart rate and subsequent periods of acardia. These decreases in rate occurred concurrently with a short-term increase in stroke volume of the heart, followed by a longer-term decrease in stroke volume. Hemolymph flow rates through the anterior aorta, anterolateral arteries, sternal artery, and posterior aorta also showed the same trend, with an initial short-term increase in flow rate followed by a longer-term decrease with periods of ischemia. Hemolymph flow through the paired hepatic arteries simply decreased, but recovery to pretreatment levels was faster than in the other arterial systems. Threshold for these responses occurred at circulating concentrations between 10(-9) mol.l-1 and 10(-8) mol.l-1 for F1 and somewhat higher, between 10(-8) mol.l-1 and 10(-7) mol.l-1, for F2.

A semi-isolated, in situ heart preparation of the shore crab, Carcinus maenas, supported by its alary ligaments, pumps vigorously for hours at a mean heart rate of 49.7 beats/min and cardiac output of 30 ml.kg-1.min-1. These hearts show no adaptive responses to changes in pericardial sinus pressure, outflow resistance, or afterload. Direct perfusion-induced stretch of the heart wall causes increases in contractile force but minimal changes in heart rate. Stroke work and power are lower than comparable values for animals with myogenic hearts and closed circulatory systems. The values for heart rate and cardiac output are lower than in vivo values and may in part reflect the technique used as well as intrinsic performance of the heart without neural and neurohormonal inputs. Morphometrically the heart represents 0.2% of whole body weight, and the mean stroke volume of 0.35-0.45 ml/kg represents an ejection fraction of 27-34% of ventricular volume (1.4 ml/kg).

Specimens of the boreal clam, Anodonta grandis simpsoniana were emersed at 10{deg}C for 6 days and then reimmersed for 24 h. The clams lost water at a rate of 1.6% total water per day. After 144 h of emersion, water weight had declined by almost 15%, while extracellular fluid (ECF) osmolality had increased 30% to 52 mOsm kg-1. Control levels were reattained after 6 h reimmersion. ECF Po2 declined rapidly in the first 24 h of emersion, but remained near 20 Torr for the full 6-day exposure. After an initial rapid fall, pH declined at a slower rate, reaching 7.494 +/- 0.037 (mean +/- SEM) at 144 h. Pco2 was elevated from 0.6 +/- 0.6 to 12.4 +/- 1.1 Torr after 96 h, but no further increase was noted. ECF [Ca] increased threefold to 13.1 +/- 0.8 mmol l-1, while [HCO3app] rose from 5.4 +/- 0.3 to a maximum of 12.9 +/- 0.8 mmol 1-1 after 144 h. ECF [Na] and [Cl] were not affected by emersion. On reimmersion, recovery was rapid, with pH, Po2 and Pco2 returning to control within 2 h, while [Ca] and [HCO3app] remained elevated until 24 h after reimmersion. A 1:1 stoichiometry between [Ca] and [HCO3app] existed throughout the emersion and reimmersion periods. In the absence of protein buffers, the fall in ECF pH was arrested by the mobilization of calcium carbonate, presumably from the shell. By 96 h emersion Pco2 and Po2 had stabilized, suggesting that diffusion gradients sufficient to allow limited gas exchange had been established.

Unrestrained, quiescent starry flounder maintained approximately normal levels of O2 uptake in the face of severe experimental anaemia. At haematocrits above about 5%, the only major compensation was a reduction in venous O2 tension which lowered venous saturation and thereby kept a constant difference between arterial and venous O2 contents. Below a haematocrit of about 5%, this difference decreased, and many additional compensations were invoked, including increases in ventilation, expired O2 tension, arterial O2 tension, and cardiac output, and decreases in systemic vascular resistance and blood pH. All changes could be reversed by restoration of haematocrit. Exercise performance and post-exercise changes in blood pH and lactate differed only slightly between anaemic and normal flounder. In wild flounder, anaemia commonly occurs and apparently only causes death at the haematocrit value (about 5%) below which most major compensations are implemented. The respiratory strategy of the flounder during anaemia is compared with that of the rainbow trout.

An isolated perfused crab gill preparation was used to test the hypotheses that crab gill carbonic anhydrase (CA) catalyzes the efflux of CO2 from the hemolymph, which lacks the enzyme, to the ambient medium and that the CA is localized on the luminal surface of the basal membrane. It was found that the efflux of CO2 from the internal perfusate was sensitive to the flow rate of the internal perfusate through the gill (and thus the residence time within the gill). The sensitivity of the CO2 efflux to residence time was nearly abolished upon treatment of the gill with an impermeable dextran-bound CA inhibitor. It is concluded that CA present on the luminal surface of the gill epithelium facilitates CO2 excretion by catalyzing the dehydration of the large hemolymph bicarbonate pool to the more diffusible molecular CO. The action of the enzyme is important in maintaining a CO2 gradient between hemolymph and water in a situation where hemolymph PCO2 is normally low, water PCO2 is variable, and the gills themselves are a source of metabolic CO2.

During swimming activity Callinectes sapidus incurred a severe hemolymph acidosis due to elevated levels of PCO2 and lactate. Although hemolymph lactate concentration rose steadily during 1 h of exercise, hemolymph pH was maximally depressed within the first 15 min. A discrepancy between the quantities of lactate and H+ released from the tissues into the hemolymph is explained by a large apparent efflux of H+ into the ambient seawater, presumably via a branchial ion exchange process. Ammonia excretion increased 6 fold during exercise, but it is not clear if this contributed to the excretion of H+. The total quantity of H+ excreted into the environment during exercise and recovery far exceeded that which could be attributed to hemolymph lactic acid, indicating that some of the excreted H+ must have originated from other sources, such lactic acid which dissociated intracellularly, with the lactate anions remaining in the cells. Because lactate anions increase hemocyanin O2 affinity when unopposed by the Bohr shift, the excretion of a large portion of the metabolic H+ load, leaving lactate behind in the hemolymph, has important consequences for the regulation of O2 transport during swimming.

A wide range of respiratory, ventilatory, and cardiovascular parameters have been recorded under completely resting conditions in the starry flounder (Platichthys stellatus), a generally inactive benthic teleost. The results differ in a number of important respects from those of a previous study on the same species. The present data have also been compared with those reported for the active pelagic rainbow trout (Salmo gairdneri) and for other teleost species. Of particular note in the flounder, relative to the trout, are low arterial and venous PO2's, a low arterial-venous O2 content difference, a low transfer factor and high diffusion gradient for O2 across the gills, a high in vivo blood O2 affinity, a high cardiac output and stroke volume accompanied by a low peripheral vascular resistance, a low ventilation volume, a low ventilation-perfusion ratio, and a low capacity-rate ratio for O2 exchange at the gills. Parameters of CO2 transport and acid-base regulation appear conventional, though blood CO2 contents and lactate concentrations are low. The respiratory strategies of inactive versus active, and benthic versus pelagic teleosts are discussed.

A whole trout preparation (Salmo gairdneri) externally ventilated with water and internally perfused with artificial medium via a cardiac pump is discribed for the study of O2 exchange and vascular resistance. As cardiac output (Q) was raised, ventral and dorsal aortic pressures increased while branchial (Rg) and systemic (Rs) vascular resistances fell, reflecting considerable passive distensibility. Arterial oxygenation was negative at low Qs due to significant internal O2 demand by the gill tissue, but increased to zero or positive values at intermediate Qs, and eventually declined at high Qs because of transit time limitation. O2 uptake from the ventilatory flow rose with increasing Q. Epinephrine (10(-5) M) decreased Rg, increased Rs, and enhanced arterial oxygenation. Artificial elevation of dorsal aortic pressure decreased Rg but did not affect arterial oxygenation. A 10-fold elevation of ventilatory flow increased arterial oxygenation but did not alter Rg or Rs. Endogenous metabolism of branchial tissue accounted for 11.7% of resting O2 uptake in vivo, and comprised an internal component taking O2 from perfusion flow and an external component drawing O2 from ventilatory flow.

A new method of calculating PCO2 based on mathematical expressions derived from a measured 'Davenport diagram' is described. The measured 'Davenport diagram' is constructed from in vitro buffer curves relating total CO2 to pH at PCO2 levels which adequately encompass the in vivo range of the acid-base status for the species in question. The 'Davenport diagram' is described by three linear equations such that PCO2 can be accurately calculated from in vivo measured CCO2 and pH. The equations are specific for a given species at a given temperature and hemolymph ionic strength, as are the constants in the more classical Henderson-Hasselbalch equation. The accuracy of the method is equal to calculations using the Henderson-Hasselbalch equation with corrected values for pK1' and alpha CO2 and in vivo PCO2 measured directly. This procedure is equally applicable to fluids with dissolved pigments such as hemolymph from freshwater and marine crustaceans and to human blood. The major benefit of this method of calculating PCO2 is that correction nomograms for the constants in the Henderson-Hasselbalch equation are not required.

Exhausting activity results in a marked and immediate drop in blood pH which gradually returns to normal over the following 6h. The acidosis is caused largely by elevated Pco2 levels, which vary inversely with pH. Blood lactate concentration increases slowly, reaching a maximum at 2--4h post-exercise, and contributes significantly to the acidosis only late in the recovery period. The slow time course of lactic acid release into the blood permits temporal separation of the peak metabolic acidosis from the peak respiratory acidosis. Evidence is presented that a metabolic acid other than lactic also makes a modest contribution to the pH depression during the recovery period.

Hemolymph samples were taken from small (< 100 g) individuals of Cancer productus following ca. 3 h air exposure (emersion) on the beach, `in situ', at Friday Harbor, Washington. Compared with crabs of similar size in sea water in the laboratory, these crabs emersed `in situ' had lower Pa o 2 , and Pv o 2 , but no significant change in pH and a small, not significant, internal hypercapnia. Total CO 2 (C c o 2 ) content of the hemolymph was elevated by 70% (15.2 versus 9.0 mM), possibly as compensation for input of acid into the hemolymph. These responses are qualitatively similar to those resulting from similar treatment in the laboratory, but differ in the reduced magnitude of the internal hypercapnia and acidosis of the hemolymph. It is suggested that the particular conditions of emersion `in situ' permit some gas exchange with interstitial sea water. Interstitial sea water was found to be hypoxic (P o 2 = 20-40 torr), which would limit oxygen supply yet permit CO 2 excretion to continue, in agreement with the data.

Hemolymph acid-base variables (pH, PCO2 and CCO2), hemolymph Ca2+ and Na+ concentrations, and osmolality were measured in unrestrained crabs, Cancer productus, before, during and following 4 hr emersion and 43 hr hyperoxia (460-510 Torr), both at 10 degrees C. Emersion and hyperoxia provoked an acidosis associated with elevation of hemolymph CCO2 and PCO2, yet attempts to calculate PCO2 from measured pH and CCO2 always resulted in values greater than those measured directly. This discrepancy between measured and calculated PCO2, was associated with base excess, and was eliminated upon in vitro equilibration of the hemolymph and more slowly in vivo, suggesting that metabolic compensation for the acidosis occurred more rapidly than could acid-base equilibration. During emersion, increases of CCO2 and [Ca2+] provide evidence that the internal CaCO3 stores, possibly from the exoskeleton, were mobilized during acid-base compensation. Hyperoxia provoked no such increase in Ca2+, and branchial uptake of HCO3- may make a major contribution to the elevation of CCO2 during hyperoxia. It is suggested that shell buffering by aquatic crustaceans provides a means of compensation for acidosis under conditions during which branchial function is impaired.