Easier threading through web-based comparisons and cross-validations.

UNLABELLED: We have developed a WWW server for the integration and comparison of protein structure predictions performed by five different servers. Users submit an amino acid sequence to a selected set of these prediction methods. Results are gathered on a web-based page in order to facilitate comparison and analysis. All the alignments are further evaluated through a common threading tool making their comparisons easier. AVAILABILITY: The meta-server is available free at http://www.infobiosud.cnrs.fr/bioserver SUPPLEMENTARY INFORMATION: http://www.infobiosud.cnrs.fr/bioserver/hah1.html

A genetic algorithm for the automated generation of small organic molecules: drug design using an evolutionary algorithm.

Rational drug design involves finding solutions to large combinatorial problems for which an exhaustive search is impractical. Genetic algorithms provide a novel tool for the investigation of such problems. These are a class of algorithms that mimic some of the major characteristics of Darwinian evolution. LEA has been designed in order to conceive novel small organic molecules which satisfy quantitative structure-activity relationship based rules (fitness). The fitness consists of a sum of constraints that are range properties. The algorithm takes an initial set of fragments and iteratively improves them by means of crossover and mutation operators that are related to those involved in Darwinian evolution. The basis of the algorithm, its implementation and parameterization, are described together with an application in de novo molecular design of new retinoids. The results may be promising for chemical synthesis and show that this tool may find extensive applications in de novo drug design projects.

Effects of catecholamines on the pulmonary venous bed in sheep.

OBJECTIVE: To assess the effects of various catecholaminergic agents on pulmonary venous tone. DESIGN: Prospective, randomized, controlled, experimental study. SETTING: Physiology laboratory of a university hospital. SUBJECTS: Thirty anesthetized, mechanically ventilated adult sheep. INTERVENTIONS: Four groups of six animals received 1-hr infusions of norepinephrine (0.5 microg/kg/min), epinephrine (0.5 microg/kg/ min), dopamine (10 microg/kg/min), or dobutamine (10 microg/kg/min). MEASUREMENTS AND MAIN RESULTS: A 7-Fr pulmonary artery catheter was placed in a proximal location to measure cardiac output and pressure in a large pulmonary vein (Ppw) after balloon inflation. Another catheter wedged in a small pulmonary artery measured pressure in a small pulmonary vein (Pdw). A third catheter measured left atrial pressure (PLA ). This method was able to detect the pulmonary venoconstrictive effects of histamine in a separate group of six animals. Pdw-PLA increased from a mean of 2.0+/-1.7 to 3.0+/-1.5 (SD) cm H2O (p < .01), 2.3+/-1.6 to 4.4+/-1.3 cm H2O (p < .01), and 1.7+/-1.0 to 3.5+/-2.2 cm H2O (p < .05) with norepinephrine, epinephrine, and dopamine, respectively. All of these drugs increased Pdw-Ppw, but only norepinephrine and epinephrine increased Ppw-PLA . No change in either pressure difference was observed with dobutamine. Elevation of cardiac output alone could not account for these findings since the increase in cardiac output induced by fluid infusion did not change the pressure differences. CONCLUSION: Norepinephrine, epinephrine, and dopamine at doses commonly used in humans increase pulmonary venous tone in sheep.

Non-invasive assessment of technetium-99m albumin transit time distribution in the pulmonary circulation by first-pass angiocardiography.

This study describes a non-invasive method for assessment of the lung transit time distribution of a tracer, using first-pass technetium-99m albumin angiocardiography and a model-free method of deconvolution. Ten patients received a first injection of 1 MBq kg-1 in the external jugular vein to position a gamma camera in the left anterior oblique position and two additional injections (5 MBq kg-1) to record first-pass angiocardiographic data. Right and left ventricular time-activity curves were derived from regions of interest every 0.5 s over a 1-min period. The left ventricular curve was deconvoluted by the right ventricular curve to obtain the lung transport function. The deconvolution procedure was based on a modified version of the Kalman filtering technique. The procedure was repeated at an interval of 30 min in eight patients. Two patients were re-examined up to 2 years later. Skewness, kurtosis and relative dispersion of the distributions did not change over time. We also found that the distribution, once normalized by its first moment, was independent of isolated changes in heart rate or cardiac output. Comparison of curve shapes at an interval of 30 min by point by point analysis demonstrated the reproducibility of the technique. We conclude that computation of the pulmonary transit time distribution of 99mTc-albumin from a standard angiocardiography procedure by model-free deconvolution is reliable and reproducible over time. We suggest that it may be a valuable tool for the non-invasive follow-up of the pulmonary circulation.

Comparison of indirect calorimetry and thermodilution cardiac output measurement in children.

We validated experimentally the ability of hood indirect calorimetry to measure accurately VO2. For this purpose we compared cardiac output calculated from the Fick equation Q = VO2/(Ca(O2) - CV(O2)), in which VO2 was obtained by hood indirect calorimetry, to thermodilution cardiac output (Qth) measured simultaneously during cardiac catheterization in children (n = 16). Because FI(CO2) is a critical factor in hood indirect calorimetry calculations, we also assessed the consequence of taking into account measured FI(CO2) rather than using the usual standard value of 0.0004. We found a good agreement between Q and Qth whether we used experimentally measured FI(CO2) in ambient air (Qth = 0.89 Q + 0.39, r = 0.941) or standard FI(CO2) (Qth = 0.84 Q + 0.55, r = 0.930). However, VCO2 and R computed from standard FI(CO2) differed significantly (p < 0.001) from values derived from measured FI(CO2). This demonstrates that indirect calorimetry allows reasonable estimates of Q, VO2, VCO2, and R provided that the actual values of FI(CO2) are used.

Chronic interleukin-2 treatment in awake sheep causes minimal or no injury to the lung microvascular barrier.

Interleukin-2 (IL-2) is reputed to cause a "vascular leak syndrome." We studied pulmonary hemodynamics and lymph dynamics in six sheep treated for 7 days with IL-2 (1.8 million IU/kg twice daily or 1.8 million IU/kg each day as a continuous infusion). Lung lymph flow increased from 4.8 +/- 2 ml/15 min pre-IL-2 to 14.4 +/- 6.8 ml/15 min on the seventh day of IL-2. The lymph-to-plasma protein concentration ratio was unchanged (0.70 +/- 0.06 vs. 0.63 +/- 0.13). The plasma-to-lymph equilibration half-time of radiolabeled albumin was 2.0 +/- 0.6 h pre-IL-2 and 1.0 +/- 0.7 h on day 7 of IL-2. Pulmonary arterial pressure was 24 +/- 7 cmH2O pre-IL-2, increased to 32 +/- 4 cmH2O on the fourth day of IL-2, and returned to 29 +/- 5 cmH2O on the seventh day of IL-2. Extravascular lung water was normal (4.07 +/- 0.25 g/g dry lung). To clearly determine whether the increase in lung lymph flow was due to hemodynamic changes or to increased leakiness of the microvascular barrier, we volume loaded six sheep with lactated Ringer solution before and after 3 days of IL-2 treatment (1.8 million IU/kg twice daily). Lung lymph flows increased fivefold during 4 h of crystalloid infusion compared with baseline and were higher after 3 days of IL-2. However, lymph-to-plasma protein concentration ratios decreased to the same low levels pre-and post IL-2 (0.39 +/- 0.06 vs. 0.41 +/- 0.10), indicating and intact microvascular barrier. Extravascular lung water was elevated (5.56 +/- 0.39 g/g dry lung) but was not different from lung water in three volume-loaded control sheep (4.87 +/- 0.53 g/G dry lung). We conclude that IL-2 causes minimal or no injury to the pulmonary microvascular barrier and that volume expansion during IL-2 treatment can cause hydrostatic pulmonary edema.

[Development of a device allowing for the perfusion and preservation of rat kidneys at -4 degrees C]. / Mise au point d'une machine permettant la perfusion et la conservation des reins de rat à -4 degrees C.

OBJECTIVES: In order to prolong the cold ischaemia time and to improve the quality of donor kidneys, we have designed and developed a renal perfusion machine allowing the control of perfusion parameters (temperature, pressure, flow rate, resistance) during the various phases of a kidney perfusion and storage protocol at -4 degrees C. ANIMALS, MATERIALS AND METHODS: Twenty four rat kidneys were removed and the effects of perfusion and storage at -4 degrees C were studied using a perfusion/storage machine allowing the controlled addition of 2,3 butanediol in University of Wisconsin (UW solution). The kidneys were stored for 96 hours at -4 degrees C and were studied in terms of perfusion parameters (pressure, resistance) and according to their histological appearance. RESULTS: The machine allows controlled perfusion of a cryoprotective agent and preservation of kidneys at -4 degrees C for 96 hours. CONCLUSION: In animals, it is possible to store kidneys at a temperature of -4 degrees C for 96 hours by using a vecor solution (UW solution) and a cryoprotective agent (2,3 butanediol). The perfusion and storage of organs under these conditions must be performed by a computer-assisted machine, allowing monitoring and control of the various steps of perfusion/storage.

Incidence of vascular complications in kidney transplantation: is there any interference with the nature of the perfusion solution?

Two non-randomized groups of 100 kidney transplants each were compared in relation to the flush-out solution used. The two groups were similar with the exceptions of the number of multiple artery kidneys and the proportion of kidneys procured in our center. The rate of vascular complications such as arterial venous thrombosis and renal artery stenosis was higher in the Eurocollins (19) group than in the UW group (4) p < 0.01, even if the multiple artery kidneys are excluded. These results need to be confirmed by a randomized study but it is clear that the use of the UW solution leads to a better arterial flow with fewer vascular complications.

Effects of interleukin-2 on the pulmonary microvasculature in anesthetized sheep.

Interleukin-2 (IL-2) is reputed to cause pulmonary microvascular injury. We studied the pulmonary and splanchnic microcirculation of anesthetized sheep after one dose (1.8 x 10(6) IU/kg) of IL-2 (n = 9) and after six doses (1.8 x 10(6) IU.kg-1.dose-1) of IL-2 over 3 days (n = 9). Seven control sheep received only 5% dextrose diluent. We measured hemodynamics and lymph dynamics in anesthetized sheep after the final dose of IL-2 or diluent. After one dose of IL-2, caudal mediastinal node (mainly pulmonary) lymph flow was stable, whereas thoracic duct lymph flow increased from a baseline of 54 +/- 6 to 124 +/- 22 ml/h. After 3 days of IL-2, the caudal mediastinal node lymph flow increased from 7.7 +/- 5.5 to 19.0 +/- 14.8 ml/h 5-6 h after the final dose of IL-2, and thoracic duct lymph flow increased from 84 +/- 43 to 143 +/- 42 ml/h. The lymph-to-plasma protein concentration ratio increased after IL-2 for thoracic duct but not for caudal mediastinal node lymph. The equilibration rate of 125I-albumin from plasma to caudal mediastinal node lymph did not change, whereas plasma-to-thoracic duct lymph equilibration was faster after both one dose and 3 days of IL-2. Positron emission tomography showed no increase in the pulmonary transcapillary escape rate for 68Ga-labeled transferrin or in extravascular lung water (n = 4). We conclude that IL-2 at doses two to three times those used clinically does not significantly injure the pulmonary microcirculation of sheep.

Study of kidney and liver viability in the rat after exclusive aortic perfusion using intracellular ATP measurement.

To find whether the liver can be procured after exclusive aortic perfusion, three organ perfusion models were used in three groups of donor rats. Group 1 underwent liver wash-out via the portal vein; in group 2, the kidneys alone were perfused via the aorta; and group 3 underwent simultaneous aortic perfusion of liver and kidneys. All perfusion flow rates in the three groups were adjusted to physiological values. Harvested organs were transplanted and recipient animals were killed 4 h after transplantation to study liver and kidney viability by using intracellular ATP measurement. Liver ATP was lower (P < 0.005) in the portal perfusion group (group 1: 1.396 +/- 0.412) than in the aortic perfusion group (group 3: 2.181 +/- 0.061). Kidney ATP was comparable in groups 2 and 3:1.066 +/- 0.09 vs 1.059 +/- 0.273 (mumol/g) tissue). Liver cooling was quicker with portal perfusion than with the aortic flush (20 degrees C in 20 s vs 15 degrees C in 60 s). Aortic perfusion at a physiologic flow rate has no detrimental effect on renal viability studied by intracellular ATP measurement. We conclude that liver cooling via the aortic route only is a good alternative to portal perfusion and seems to give good preservation. Application of this observation to emergency procurement in humans is still the subject of controversy.

Muscular venous blood metabolites during rhythmic forearm exercise while breathing air or normoxic helium and argon gas mixtures.

The effects of breathing normoxic helium or argon gas mixture on the local muscular metabolism during exercise were compared to those of room air in four healthy subjects. For this purpose, PO2, PCO2, pH, the concentrations of lactate (LA), glucose (Gl) haemoglobin (Hb) and K+ and osmolarity were repeatedly measured in efferent muscular venous blood during 12 min of rhythmic forearm exercise and 16 min of recovery. The time courses and magnitude of the changes in PCO2, pH, [Gl], [Hb], [K+] and osmolarity during exercise and recovery were similar for breathing both the helium and argon gas mixtures. The main finding was that during exercise, the [LA] curve reached a peak value significantly higher by 25% under normoxic helium than under room air or normoxic argon breathing. This rise in [LA] was accompanied by a slight reduction in muscular venous PO2, too small to signify muscular hypoxia. The possibility that this decline in PO2 might be due to a shift in muscular metabolism towards lipid oxidation was confirmed by the lower muscular respiratory quotient observed during helium breathing. However, such a shift did not explain why [LA] rose during this breathing. The probable explanation is that helium facilitates LA diffusion out of the myocytes.