Functioning of the electromechanical connection in the course of the contracture contraction

The effects of calcium release blocker dantrolene was tested on electrically evoked twitches and on contractures induced by potassium depolarization, by acetylcholine or caffeine. It was shown that the first: developmental, stage of potassium or acetylcholine contracture is inhibited by dantrolene and is not influenced by calcium free medium, therefore we may interpret it as based on a "voltage-dependent Ca release" (VDCR) mechanism of activation, whereas depolarization directly opens the rhyanodin receptor calcium channels. On the contrary, the next stage: the long-lasting plateau of contracture, is directly dependent on external Ca2+ and inhibited by dantrolene, and therefore can be described as "calcium induced Ca-release" (CICR) activation mechanism. In this case stored calcium is also released by rhyanodine receptors, although by means of entering the extracellular Ca2+. Finally, the last stage of low amplitude is not influenced by dantrolene nor by calcium-free medium. Therefore the activation of contraction on this stage is not based on the Ca2+ release through the rhyanodin receptor calcium channels.     

The innervation of the closer muscle of the mesothoracic spiracle of the locust.

The closer muscle of the mesothoracic spiracle of the locust, Schistocerca gregaria is innervated by two excitatory motoneurones and also by processes of a peripherally located neurosecretory cell. Within the muscle, ultrastructural studies show the presence of two types of excitatory nerve terminal which differ in the content of dense cored vesicles and in their distribution. The ventral segment of the muscle is innervated predominantly by terminals with small clear vesicles and only an occasional dense-cored vesicle. The central part of the muscle is innervated predominantly by terminals with small clear vesicles and larger numbers of dense-cored vesicles. The dorsal segment of the muscle is innervated exclusively by a neurosecretory type innervation. The small neurohaemal organ of the median nerve close to the spiracle muscle is immunoreactive to an antibody raised against bovine pancreatic polypeptide but no immunoreactive processes enter the muscle itself. The muscle possesses specific octopaminergic receptors that increase cyclic AMP levels and the possibility that the neurosecretory input to the muscle is provided by either a central or peripheral octopamine containing neurone is discussed.     

Characteristics of the membranes of the pellicle of the ciliatePseudomicrothorax dubius

Summary The membranes of the pellicle of the ciliatePseudomicrothorax dubius are investigated using thin section electron microscopy and freeze-fracture replicas. The plasma membrane is covered by a surface coat and is connected to the outer alveolar membrane by short, sometimes branched, bridges. The inner alveolar membrane is coated on both sides. The epiplasm lies in intimate contact with the cytoplasmic surface of this membrane, and there is a corresponding deposit on the other surface. This deposit is regularly striated.The epiplasmic layer and the alveoli are interrupted at sites of cytotic activity,e.g., the attachment sites of trichocysts, the cytoproct, and the parasomal sacs. The striated deposit ends where the epiplasm ends, indicating a direct relationship between these two epimembranous layers.There is a deposit along the sides of the first part of the tip of the trichocysts, and in this region the trichocyst membrane is free of intramembranous particles.The membrane of the parasomal sacs has a coat on both surfaces. That on the extraplasmic surface is similar to the surface coat of the plasma membrane. The origin of the cytoplasmic coat is unknown. The cytotic activity of these sacs is indicated by their highly irregular profiles.     

A comparison of the properties of the pyruvate kinases of the fat body and flight muscle of the adult male desert locust

1. The pyruvate kinases of the desert locust fat body and flight muscle were partially purified by ammonium sulphate fractionation. 2. The fat-body enzyme is allosterically activated by very low (1mum) concentrations of fructose 1,6-diphosphate, whereas the flight-muscle enzyme is unaffected by this metabolite at physiological pH. 3. Flight-muscle pyruvate kinase is activated by preincubation at 25 degrees for 5min., whereas the fat-body enzyme is unaffected by such treatment. 4. Both enzymes require 1-2mm-ADP for maximal activity and are inhibited at higher concentrations. With the fat-body enzyme inhibition by ADP is prevented by the presence of fructose 1,6-diphosphate. 5. Both enzymes are inhibited by ATP, half-maximal inhibition occurring at about 5mm-ATP. With the fat-body enzyme ATP inhibition can be reversed by fructose 1,6-diphosphate. 6. The fat-body enzyme exhibits maximal activity at about pH7.2 and the activity decreases rapidly above this pH. This inactivation at high pH is not observed in the presence of fructose 1,6-diphosphate, i.e. maximum stimulating effects of fructose 1,6-diphosphate are observed at high pH. The flight-muscle enzyme exhibits two optima, one at about pH7.2 as with the fat-body enzyme and the other at about pH8.5. Stimulation of the enzyme activity by fructose 1,6-diphosphate was observed at pH8.5 and above.     

Cytochemical study of the distribution of adenosine triphosphatase in the pancreas of the dog.

Dog pancreatic tissue, incubated in a modified Wachstein-Meisel medium, showed two different adenosine triphosphatase activities. One of them is located at the apical border of the cells lining the intralobular ducts and of the centroacinar cells and is stimulated by HCO3-, depressed by SCN- and OCN- and completely abolished by CN-. The other is located at the intracellular clefts of the epithelium lining the interlobular ducts and is stimulated by Mg++. These findings correlate well with the results of incubation of homogenates of fresh and fixed tissues. Their significance with respect to the role of different segments of the duct system in the formation of the pancreatic juice is discussed.     

Functional morphology of the subgenual organ of the carpenter ant

Using light microscopy, confocal microscopy, electron microscopy and histochemistry, the subgenual organ (SGO) of an ant, Camponutas ligniperda, is investigated. Sensory units and attachment cells together enclose a large extracellular cavity, which is filled by acid mucopolysaccharides, as revealed by staining with ruthenium red. Due to this cavity, the whole SGO has the shape of a deformed sphere and the scolopidia exhibit a distribution of angles between 0 degrees and 60 degrees with the tibial long axis (as is shown by phalloidin-rhodamin staining of the actin filaments of the scolopale, viewed in situ by laser scanning confocal microscopy). The subgenual organ is innervated by a branch of the tibial nerve, which splits within or shortly distal to the femur-tibia joint. The other features of the SGO of Camponotus ligniperda are similar as in other insects: the SGO of Camponotus ligniperda contains about 35 scolopidial sensilla; it is fixed to the subgenual nerve on its proximal end, by its attachment cells to the opposite part of the cuticle; the fixation by the attachment cells is accomplished by a vast quantity of cytoplasmic microtubules; the construction of the sensory units is the same as in other mononematic scolopidial organs. The role of the extracellular lumen inside the organ and the special shape of the SGO of Camponotus ligniperda in mechanical transmission is discussed.     

Control of the circadian rhythm of the body temperature in the rat

The circadian rhythm of the rat body temperature is abolished in rats kept either in constant dark or light or blinded. The suppressive effects act most likely via the retinas. The rhythm can be inverted by reversal of the lighting regimen and it is unaffected by pinealectomy. Most likely the circadian rhythm is not related to changes in patterns of motor activity.     

Role of the temperature factor in determining the nature of the chronotropic response of the heart of cats in the simulation of increased venous inflow

The heart rate changes in response to the imitation of venous input to the heart by infusion of an additional blood volume to the right atria have been studied on anesthetized by chloralose cats. It has been discovered that patterns of the heart rate changes are determined by the infused blood temperature if it is equal to the blood temperature in heart, tachycardia appears, if it is below this temperature, bradycardia develops. These data explain, from our point of view, the contradictory results obtaining by different authors in experiments with the imitation of the increased venous input (Bainbridge reflex).     

A clinico-physiological study of the mechanism of the cholecystokinetic action of medicinal mineral waters of the naftusia type]

It is stated that the cholecystokinetic action of curative waters of naftusia type is due to the interaction of its organic substances, particularly bitumens, with mucous membrane of gastroduodenal zone by the intramural reflex mechanism with participation of humoral factor. It is indicated by attenuation or suppression of the effect by pharmacons interrupting the work of interchemoreception, N- and M-cholinoreceptors, beta-adrenoreceptors; intensification of the effect by blockade of alpha-adrenoreceptors; its reversion by nonselective stimulation of adrenoreceptors; release of gastrin, glucagon and insulin into blood.     

Probing the role of the hyper-reactive histidine residue of arginase

Rat liver arginase (arginase I) is potently inactivated by diethyl pyrocarbonate, with a second-order rate constant of 113M(-1)s(-1) for the inactivation process at pH 7.0, 25 degrees C. Partial protection from inactivation is provided by the product of the reaction, l-ornithine, while nearly complete protection is afforded by the inhibitor pair, l-ornithine and borate. The role of H141 has been probed by mutagenesis, chemical modulation, and X-ray diffraction. The hyper-reactivity of H141 towards diethyl pyrocarbonate can be explained by its proximity to E277. A proton shuttling role for H141 is supported by its conformational mobility observed among the known arginase structures. H141 is proposed to serve as an acid/base catalyst, deprotonating the metal-bridging water molecule to generate the metal-bridging hydroxide nucleophile, and by protonating the amino group of the product to facilitate its departure.     

Reversal of the effect of the allosteric ligands of dCMP-aminohydrolase and stabilization of the enzyme in the T form

The 5-mercury derivative of dCMP is a substrate of deoxycytidylate aminohydrolase in the presence of mercaptoethanol. With this substrate a reversal of the effect of the allosteric ligands of the enzyme is observed. dCTP, which is an allosteric activator for aminohydrolysis of dCMP, becomes an inhibitor for the mercury substrate, whilst dTTP, an allosteric inhibitor for dCMP, becomes an activator for the mercury substrate.This observation has been interpreted by assuming that dCMP-Hg-S-CH₂-CH₂-OH is a substrate of the T form of the enzyme. By reacting dCMP-aminohydrolase in the T form (in the presence of dTTP) with glutaraldehyde, an enzyme has been isolated that is no longer active with dCMP, while it is fully active with the mercurated analog. Gel electrophoresis demonstrated that glutaraldehyde does not produce intermolecular crosslinks, but fixes 95% of the enzyme in a stable hexameric form by intramolecular crosslinks. The data are explained by assuming that glutaraldehyde stabilizes the enzyme in the T conformation.     

Exploring the origin of the ion selectivity of the KcsA potassium channel

The availability of structural information about biological ion channels provides an opportunity to gain a detailed understanding of the control of ion selectivity by biological systems. However, accomplishing this task by computer simulation approaches is very challenging. First, although the activation barriers for ion transport can be evaluated by microscopic simulations, it is hard to obtain accurate results by such approaches. Second, the selectivity is related to the actual ion current and not directly to the individual activation barriers. Thus, it is essential to simulate the ion currents and this cannot be accomplished at present by microscopic MD approaches. In order to address this challenge, we developed and refined an approach capable of evaluating ion current while still reflecting the realistic features of the given channel. Our method involves generation of semimacroscopic free energy surfaces for the channel/ions system and Brownian dynamics (BD) simulations of the corresponding ion current. In contrast to most alternative macroscopic models, our approach is able to reproduce the difference between the free energy surfaces of different ions and thus to address the selectivity problem. Our method is used in a study of the selectivity of the KcsA channel toward the K+ and Na+ ions. The BD simulations with the calculated free energy profiles produce an appreciable selectivity. To the best of our knowledge, this is the first time that the trend in the selectivity in the ion current is produced by a computer simulation approach. Nevertheless, the calculated selectivity is still smaller than its experimental estimate. Recognizing that the calculated profiles are not perfect, we examine how changes in these profiles can account for the observed selectivity. It is found that the origin of the selectivity is more complex than generally assumed. The observed selectivity can be reproduced by increasing the barrier at the exit and the entrance of the selectivity filter, but the necessary changes in the barrier approach the limit of the error in the PDLD/S-LRA calculations. Other options that can increase the selectivity are also considered, including the difference between the Na+...Na+ and K+...K+ interaction. However, this interesting effect does not appear to lead to a major difference in selectivity since the Na+ ions at the limit of strong interaction tend to move in a less concerted way than the K+ ions. Changes in the relative binding energies at the different binding sites are also not so effective in changing the selectivity. Finally, it is pointed out that using the calculated profiles as a starting point and forcing the model to satisfy different experimentally based constraints, should eventually provide more detailed understanding of the different complex factors involved in ion selectivity of biological channels.     

Electron microscopical study on the development of the nerve supply of the pituitary pars intermedia of the mouse

Summary The development of the nerve supply of the pituitary pars intermedia (PI) of C3H mice was studied by electron microscopy. Nerve fibres and terminal structures, most probably adrenergic, first appear in the newborn. The adult innervation pattern is achieved by the end of the first postnatal week.In the adult animal two types of nerve terminals were distinguished; type A (peptidergic or neurosecretory) and type B (adrenergic). The peptidergic fibres were scarce and exhibited no synapse-like contacts. It is suggested that they are of secondary importance in a direct nervous hypothalamic control of PI function. Type B terminals were found throughout the PI. They formed synapse-like contacts with the glandular cells, indicating that the primary innervation is exerted by adrenergic neurons.An autonomous differentiation of the glandular cells and in the adult a combined direct nervous and neurohumoral control of PI function is suggested.This investigation was supported by grant No B 2180-026 from the Swedish Natural Science Research Council. The skilful technical assistance of Mrs Ulla Wennerberg is gratefully acknowledged     

Principles of the structuro-functional organization of the hemomicrocirculatory bed of the intramural nervous plexuses of organs of the gastrointestinal tract of mammals

The work deals with one of the least elaborated aspects on the system of microcirculation and is performed, taking into account the functions of histo-hematic and intertissue barriers. For this certain new original investigation methods have been developed, adequate to the problems discussed. The module principle of organization with a distinct functional specialization of the ganglionar and truncal vascular micromodules has been stated. The working principle of each module is based on functional interaction of the intra- and extraneural capillary systems; this is ensured by mechanisms of the blood stream redistribution: by arteriolar, precapillary and prevenular sphincters and by pathways of the shunt blood stream. A high stability of the blood stream in the vascular constructions studied is explained as a possibility of counterstream gas exchange between the arterial and venous blood in the truncal vascular micromodule.     

Stochastic simulation of the gas diffusion in the air phase of the human lung

The solution of the diffusion equation in the gas phase of the human lung is very difficult because of the structure of the bronchial tree. It is shown by means of physical arguments, how one can reduce the diffusion equation to a simple one-dimensional form. The solution is then obtained by a stochastic simulation, which is easily realized on a digital computer.     

Matching the supply of bacterial nutrients to the nutritional demand of the animal host

Various animals derive nutrients from symbiotic microorganisms with much-reduced genomes, but it is unknown whether, and how, the supply of these nutrients is regulated. Here, we demonstrate that the production of essential amino acids (EAAs) by the bacterium Buchnera aphidicola in the pea aphid Acyrthosiphon pisum is elevated when aphids are reared on diets from which that EAA are omitted, demonstrating that Buchnera scale EAA production to host demand. Quantitative proteomics of bacteriocytes (host cells bearing Buchnera) revealed that these metabolic changes are not accompanied by significant change in Buchnera or host proteins, suggesting that EAA production is regulated post-translationally. Bacteriocytes in aphids reared on diet lacking the EAA methionine had elevated concentrations of both methionine and the precursor cystathionine, indicating that methionine production is promoted by precursor supply and is not subject to feedback inhibition by methionine. Furthermore, methionine production by isolated Buchnera increased with increasing cystathionine concentration. We propose that Buchnera metabolism is poised for EAA production at certain maximal rates, and the realized release rate is determined by precursor supply from the host. The incidence of host regulation of symbiont nutritional function via supply of key nutritional inputs in other symbioses remains to be investigated.     

New insights into the regulation of plant succinate dehydrogenase. On the role of the protonmotive force

Regulation of succinate dehydrogenase was investigated using tightly coupled potato tuber mitochondria in a novel fashion by simultaneously measuring the oxygen uptake rate and the ubiquinone (Q) reduction level. We found that the activation level of the enzyme is unambiguously reflected by the kinetic dependence of the succinate oxidation rate upon the Q-redox poise. Kinetic results indicated that succinate dehydrogenase is activated by both ATP (K(1/2) approximately 3 microm) and ADP. The carboxyatractyloside insensitivity of these stimulatory effects indicated that they occur at the cytoplasmic side of the mitochondrial inner membrane. Importantly, our novel approach revealed that the enzyme is also activated by oligomycin (K(1/2) approximately 16 nm). Time-resolved kinetic measurements of succinate dehydrogenase activation by succinate furthermore revealed that the activity of the enzyme is negatively affected by potassium. The succinate-induced activation (+/-K(+)) is prevented by the presence of an uncoupler. Together these results demonstrate that in vitro activity of succinate dehydrogenase is modulated by the protonmotive force. We speculate that the widely recognized activation of the enzyme by adenine nucleotides in plants is mediated in this manner. A mechanism that could account for such regulation is suggested and ramifications for its in vivo relevance are discussed.     

Perinatal development of the small intestine of the sheep

Soon after birth, the villi in distal regions of the small intestine of the sheep rapidly decrease in size so that by day 6 the normal proximal-distal size gradient (tallest villi proximally) is established. This is the reverse of the gradient found throughout fetal life, where distal villi are taller than proximal villi. While crypt depth increases in both regions, the increase is more marked in proximal regions. The proportion of crypt cells labelled with a pulse of (3H) thymidine increases dramatically during day 1. Vacuolated cells are present in distal regions at day 2, but are absent by day 5. It is possible that the rapid loss of villus tip cells in distal regions is the means by which closure is achieved in this species. There is evidence to suggest that endogenous cortisol plays an important role in these changes.