Michaelis complexes of porcine pancreatic elastase with 7-[(alkylcarbamoyl)amino]-4-chloro-3-ethoxyisocoumarins: translational sampling of inhibitor position and kinetic measurements.

A step leading to the formation of the covalent complexes between porcine pancreatic elastase (PPE) and 7-[(alkylcarbamoyl)amino]-4-chloro-3-ethoxyisocoumarins (alkylHNCO-EICs) is the formation of the noncovalent Michaelis complex. No average structures are available for the Michaelis complexes of PPE with alkylHNCO-EICs. We present the results of an initial step in obtaining these structures and have determined kinetic constants as well. The kinetic results indicate that formation of the Michaelis complex is what differentiates the effectiveness of these inhibitors in inactivating PPE. The structural and kinetic results together suggest that the structure of the Michaelis complex is necessary for the design of potent alkylHNCO-EIC inhibitors of PPE. Two novel alkylHNCO-EICs are predicted to be the best inhibitors of this series. An alternate mechanism for serine protease inhibition is also proposed. Evidence for, and studies that may add support to, the hypothesized mechanism are discussed.     

An experimental test of the eddy correlation technique over a Mediterranean macchia canopy

Abstract. Flux densities of water vapour and carbon dioxide were measured for a Mediterranean macchia canopy. Results show good agreement between the measured available energy and the sum of latent sensible and heat flux densities determined with the eddy correlation technique. Joint evaluation of the Bowen ratio, aerodynamic resistance, canopy resistance and the 'omega factor' suggests that the macchia canopy is intermediate in aerodynamic roughness between coniferous and deciduous canopies. Maximum daytime carbon flux densities ranged from -14 to -22(μnol m⁻² s⁻¹ on a ground area basis. The ratio of transpiration to assimilation (E/A) was a function of incident photo-synthetic photon flux density below about 400 μmol m⁻²s⁻¹ and above it was fairly constant at 272 mol mol⁻¹ (H₂O/CO₂). The relationship between carbon influx and canopy conductance was linear. Results show promising applications of the eddy correlation technique for evaluating physiological features of canopies, treated as unitary functional systems.     

Effects of different rates of cardiac pacing on rat myocardial energy status

The energy status of mammalian cells is a finely regulated phenomenon. This is especially true in cardiac muscle cells in which energy requirements are high and the system must provide rapid turnover of the adenine nucleotides and instant response to changes in energetic demands. We have examined the acute response of the rat myocardium to ventricular pacing up to 2.5 times the resting heart rate. The purpose of this study was to determine at what level of pacing the normal energy status could be maintained and at what point it was compromised. Myocardial energy charge (EC = (ATP + 0.5 ADP)/(ATP + ADP + AMP)) was maintained at 1, 1.5 and 2 times the resting heart rate but declined significantly at 2.5 times. In contrast, phosphorylation potential (PP = ATP/ADP₁ × Pi) was drastically altered in hearts paced at 1.5, 2 and 2.5 times the resting rate. Tissue lactate increased and glycogen decreased in a linear fashion as pacing rate increased, indicating that the metabolic challenge was proportional to the pacing rate. EC seems to reflect the overall status of the cell and its ability to maintain a dynamic equilibrium. PP may reflect the immediate and necessary driving force for mitochondrial respiration in times of increased demand. These data suggest that the myocardium may meet the increased energy demands of acute ventricular pacing by shifting the molar ratio of ATP to ADP times Pi in favour of driving phosphorylation.     

Searching behaviour and cereal aphid consumption by Bembidion lampros and Pterostichus cupreus, in relation to temperature and prey density

Maximum consumption rates were determined for two carabids, Bembidion lampros Herbst. and Pterostichus cupreus L., feeding on the cereal aphid Rhopalosiphum padi L. at different temperatures in the laboratory. Mean daily consumption increased with increasing temperature for both species, B. lampros consuming a maximum of 16 1–3 instar nymphs and 9 apterous adult aphids at 25°C. P. cupreus was particularly voracious and consumed 125 apterous adult R. padi per day at 20°C. The behaviour of both species was analysed by video filming starved beetles, maintained at different constant temperatures, in arenas sown with spring barley. The behavioural components (1) still; (2) run/walk; (3) search and (4) confrontation were identified and were common to both species. P. cupreus was more active over the temperature range tested; B. lampros was inactive under 10°C. The proportion of time spent searching, number of plants searched, and velocity increased with increasing temperature for both species. When observed in similar arenas seeded with R. padi colonies, individuals of P. cupreus significantly increased their time spent searching in arenas with increasing aphid density. Following discovery of an aphid colony, individuals climbed and searched the host plant and its nearest neighbours. Plants in aphid free arenas were rarely climbed. B. lampros was not observed climbing in either aphid free arenas or in arenas with increasing aphid densities, and did not significantly increase its time spent searching in response to increased prey density. The few B. lampros that found aphids caught them walking on the soil surface. The relative efficiences of these two carabids as predators of R. padi are discussed, and the results are compared with similar studies elsewhere with predators of Sitobion avenae on winter wheat.

---

Résumé Les taux maximum de consommation de R. padi L. à différentes températures ont été déterminés au laboratoire chez deux carabes, B. lampros Herbstet P. cupreus L. La consommation moyenne a augmenté avec la température chez les deux espèces, B. lampros consommant un maximum de 15,9 larves des stades 1 à 3 et 9,1 pucerons adultes aptères, à 25°C. P. cupreus a été particulièrement vorace et a consommé 125,3 adultes aptères par jour à 20°C. Le comportement des deux espèces a été observé en filmant en vidéo des carabes à jeun, maintenus à différentes températures constantes, dans des enceintes semées en orge de printemps. Des éléments du comportement, communs aux deux espèces, ont été définis: 1) immabilité, 2) marche et course, 3) recherche, 4) affrontement. P. cupreus a été plus actif à toutes les températures, B. lamprosa été inactif au-dessous de 10°C. La part de temps consacrée à la recherche, le nombre de plantes prospectées, et la vitesse ont augmenté avec la température chez les deux espèces. Dans des enceintes similaires colonisées par R. padi, P. cupreus a significativement augmenté le temps consacré à la recherche dans les enceintes, parallèlement à l'augmentation de la densité des pucerons. Après la découverte d'une colonie de pucerons, P. cupreus escalade et prospecte la plante et ses voisines immédiates; tandis que les plantes des enceintes sans pucerons sont rarement escaladées. B. lampros n'a pas été observé escaladant des plantes d'enceintes avec ou sans pucerons, et il n'a pas accru son temps de prospection en fonction de la densité de pucerons. Les quelques B. lampros qui ont capturé des pucerons l'ont fait lorsque ceux-ci marchaient sur la surface du sol. La discussion a porté sur l'efficacité relative des deux carabes comme prédateurs de R. padi, et les résultats ont été comparés à ceux d'études du même type, menées ailleurs, avec des prédateurs de Sitobion avenae sur blé d'hiver.

     

Respiration Rate of Steinernema feltiae Infective Juveniles at Several Constant Temperatures

Respiration was measured in dauer stages of the insect-parasitic nematode Steinernema feltiae (= Neoaplectana carpocapsae) at 7, 17, and 27 C. Respiration, Q₁₀, and nematode viability were temperature dependent. Mean O₂ consumption for 5 × 10⁵ nematodes the first 24 hours was 0.27 ml at 7 C, 0.83 ml at 17 C, and 2.68 ml at 27 C. The Q₁₀ was 3.10 for 7-17 C and 3.24 for 17-27 C. Some nematodes died during 2, 14, and 21 days at 27, 17, and 7 C, respectively. The respiratory quotient was below 1 at all temperatures tested. A standard asymptotic model is expressed as oxygen consumed = 2.77 * {1 - exponent[-time * exponent(-B + C * temperature)]}; where 2.77 is the maximum response at 27 C. This model estimates nematode O₂ consumption and viability at storage temperatures between 7 and 27 C. The nematodes died when the O₂ concentration reached 0.5 ml/5 × 10⁵ nematodes. This model may be used to predict O₂ requirements of S. feltiae infective juveniles when stored as a waterless concentrate.     

低温对大鼠骨骼肌能量代谢的影响——Ⅰ.高能磷酸化合物含量的变化

本实验选择对低温比较敏感的骨骼肌组织作为实验对象。将19只大鼠随机分成3组,其右后肢经冷冻处理使组织温度分别达 5℃、0℃及-5℃,于复温后不同时间测定骨骼肌中四种高能磷酸化合物AMP,ADP、ATP及CP的含量。结果表明,大鼠骨骼肌组织中高能磷酸化合物含量随冷冻时组织最低温度的下降而不同程度的减少,并且具有明显的时相过程。复温后即刻至4小时下降迅速,4～12小时三组值较为接近,其后随时间延长而明显分离。 5℃组恢复得早而且明显,0℃组恢复程度次于 5℃组,-5℃组含量下降最明显而且几乎不恢复。提示:低温使骨骼肌组织能量的产生和贮存过程受到影响。     

The trajectory of a stiff rod in a curved potential energy trough

The equilibrium trajectory of the axis of a rod subject to an externally imposed curved potential energy trough tends to conform to the shape of the curved trough, but also tends to be straight because of elastic resistance to bending. The actual path of the axis is a balance between the two extremes. We consider a potential energy trough centered along a circular arc of radiusR. For a rod of small length compared toR, we show that the axis at equilibrium forms an arc of a circle of radius greater thanR. The value of the radius of the axial path depends on the relative values of the Hooke’s Law bending constant for the rod and the depth and width of the trough. Motivation for the calculation is provided by nucleosomal DNA, which conforms to the surface of a roughtly cylindrical histone core at physiological ionic strength, but is observed to unwind into a partially extended conformation at very low ionic strength. We suggest that the rigidity to bending of short DNA segments becomes sufficiently great at low ionic strength to overcome attractive interactions with the histone surface. Alternately, of course, if during the cell cycle mutually attractive forces between DNA and histone core are weakened at constant ionic strength, the same type of unfolding would be expected to occur as the strength of the DNA-histone contacts drops below the level required to overcome elastic resistance to bending of the DNA rod.     

The semiquinone cycle. A hypothesis of electron transfer and proton translocation in cytochromebc-type complexes

The Q cycle and theb cycle are the main current models of action of the cytochromebc-type complexes of mitochondria, bacteria, and chloroplasts. Both are based on the concept, proposed in 1972, of two sequential one-electron oxidations of (ubi)quinol along two discrete pathways which operate at different redox potentials, and with bound semiubiquinone as an intermediate. The models differ in two respects, viz. in the pathway of electron transfer and the principle of linkage of electron transfer to proton translocation. In this article we outline a new model, called the semiquinone or, simply, SQ cycle, which is based on the electron transfer principles of theb cycle but which incorporates the Q cycle concept of direct coupling between electron transfer and proton translocation through action of ubiquinone.This paper is dedicated to the memory of Bob Casey, who died in Helsinki on the 2nd of August 1985.     

Growth and production of O+ bream (Abramis brama), O+ roach (Rutilus rutilus) and O+ carp (Cyprinus carpio) in 10 drainable 0.1 Ha ponds

In ten 0.1 ha drainable ponds O⁺ cyprinids were stocked to assess their impact on the aquatic community. Bream and roach were stocked in early June, 1986, and carp in early July, 1986. The ponds were drained mid November, 1986. The relationship between the fish community and its food resource was estimated in terms of production and consumption with use of the data on growth and mortality of the fish. The average gross production of the fish species in the ten ponds (48.4 kg during the experiment) was dominated by the carp (58.5%). A minimum estimate of the food consumption of the O⁺ cyprinids is 760 kJ.m^–2. The average production of bream and roach was much lower than that of carp due to a higher mortality and a more specialized feeding pattern. The gross productions of carp, bream and roach in the ten ponds were not significantly related, although stomach analysis showed diet overlap between the carp and the bream.     

Structural basis of hierarchical multiple substates of a protein. II: Monte Carlo simulation of native thermal fluctuations and energy minimization

Conformational fluctuations in a globular protein, bovine pancreatic trypsin inhibitor, in the time range between picoseconds and nanoseconds are studied by a Monte Carlo simulation method. Multiple energy minima are derived from sampled conformations by minimizing their energy. They are distributed in clusters in the conformational space. A hierarchical structure is observed in the simulated dynamics. In the time range between 10(-14) and 10(-10) seconds dynamics is well represented by a superposition of vibrational motions within an energy well with transitions among minima within each cluster. Transitions among clusters take place in the time range of nanoseconds or longer.