Effect of etimizol on instrumental learning in rats

The action of etimizol on the acquisition of the conditioned reflexes with different complexity and biological significance of reinforcement was studied. The acquisition was performed in a shuttle box and U-shaped maze using food and footshock stimuli. The time interval between administration of etimizol (3 mg/kg) and the onset of learning varied between 0.5 and 3 h in the several series. Etimizol did not facilitate the learning in rats whatever the time of administration and biological modality of reinforcement. It is suggested that the positive effect of etimizol on the memory is related to its influence on the consolidation stage.     

Etimizol absorption from the small intestine in dogs: the dependence on dosage

As etimizol has been previously reported to be of low bioavailability, the kinetics of its absorption from the dog intestinal loop was investigated in chronic experiments. After administration of etimizol at doses of 10 or 1 mg/loop mean residence time of etimizol in the loop was 20.1 and 7.6 min, respectively, with mean standard deviation being 3.1 and 0.8, respectively. It was concluded that overall etimizol absorption from small intestine was high and its low bioavailability resulted from presystemic drug metabolism, as well as that the absorption kinetics was dose-dependent, with saturation reached at higher drug doses.     

Use of biological methods in the analysis of the purinergic activity of etimizol

To analyse etimizol purinergic activity, methods of biological analysis were applied using isolated smooth muscle strips of the guinea-pig caecum and developing embryo cultures of the sea urchin (Strongylocentrotus intermedius). Stimulating etimizol action in 10(-8)-10(-12) M concentration was observed on embryonal development in the sea urchin to be eliminated by co-action of etimizol and P1-purinoceptor antagonist theophylline. The analogous action of the preparations was established using isolated smooth muscle strips of the guinea-pig caecum.     

Effect of trisodium citrate treatment on hybridoma cell viability

Trisodium citrate has been widely used to dissolve calcium alginate beads in order to estimate cell concentration in the beads. To obtain an accurate measurement of viable cell concentration in calcium alginate beads, the effect of trisodium citrate solutions on hybridoma cell viability was studied with regard to stage of growth and trisodium citrate concentration. The cells in the decline phase of growth were more sensitive to 30 minutes of trisodium citrate treatment than the cells in the exponential phase of growth. The cell viability did not decrease rapidly during citrate treatment regardless of cell growth phase and trisodium citrate concentration in the range of 1–1.5%. By using the commercially available sodium alginate, Keltone LV, dissolution time can be kept short enough (below 5 minutes) to keep the effect of trisodium citrate negligible to cell viability.     

Reproduction of "passive" avoidance in rats by administering pharmacologic agents

In experiments on rats learned to passive avoidance reaction in one trial with subsequent administration of electroconvulsive shock (two hours after learning), the influence of different factors on the retrieval of the lost reaction was tested after three days. The greatest restoring capacity was exhibited by a non-specific reminding agent, the bell, gamma-amino-butyric acid (200 mg/kg) and etimizol (1.5 mg/kg). In animals with a preserved reaction, a number of pharmacological agents impaired retrieval of the habit (caffeine 5 mg/kg, carbocholine 0.01 mg/kg and metamizil 0.5 mg/kg). Optimal conditions for the restoration of the lost reaction were formed by etimizol and gamma-aminobutyric acid. Cholinergic mechanisms play a certain role in the functioning of the retrieval apparatus.     

The Time Effect of Auxin and Calcium on Growth and Elastic Modulus in Hypocotyls

The relationship between Young's modulus and longitudinal growth has been studied on growing segments of etiolated sunflower hypocotyls. The modulus was determined by means of the resonance frequency method. IAA in a concentration of 2.8 10^?5 M produces a decrease in the modulus with a time lag of 4 minutes, while an increase in growth is observable only after 6 minutes. Addition of IAA results in a stronger decrease in the modulus if the segments are placed in a solution of 0.1 M mannitol rattier Hum in water. Through plasmometric measurements it has been found that the elastic extensibility is insignificant compared with the growth. After the addition of IAA, there occurs a marked elongation both in 0.1 M mannitol and in water, and at the same time a decrease in the elastic extensibility of the segments is observed. In the growing segments an increased modulus causes an in creased elastic extensibility, a matter which is directly contrary to the relationship found in a physical system with an applied external force. An explanation of this discrepancy has been given. With an excess of calcium the modulus increases, while the elongation decreases. Calcium deficiency implies both a decreased modulus and a decreased growth. With the addition of 10^?3 M Ca(NO₃)₂ to segments raised without calcium the modulus increases after only 10 minutes, while an increase in longitudinal elongation is observable after 30 minutes. With the addition of IAA to the calcium deficient segments the modulus decreases to the same extent as in an optimal supply o f calcium. The results are discussed with reference to other investigations about elasticity and growth. It has been concluded that plastic extensibility cannot he of great importance in longitudinal growth. Time studies of the auxin effect and I he interaction between auxin and calcium have confirmed the hypothesis that one of the primary actions of auxin consists in a loosening of the cell wall matrix. Calcium always increases Youngs modulus and gives the cell wall a more rigid structure. Furthermore, calcium in a certain concentration is necessary for longitudinal growth.     

The Effects of Zinc on Contractility,Membrane Potentials,and Cation Content of Rat Atria

Zinc depresses the contractile force of electrically driven rat atria logarithmically with time. The threshold concentration is about 5 x 10^-6 M zinc and the half-time for contractile depression at 10^-4 M is about 25 minutes. Zinc also depresses spontaneous activity of atria and alters the transmembrane potential parameters in a manner similar to quinidine. Unlike quinidine, zinc causes an elevation of the resting potential and an elevation of cellular potassium which varies with time in the same way as the resting potential. Exposure to 10^-4 M zinc for 60 minutes causes a statistically significant fall in atrial calcium content and an amount of radioactively labeled zinc is taken up which is quantitatively equal to the calcium lost. Zinc has no effect on rigor caused by iodoacetate but inhibits rigor caused by 1-fluoro-2,4 dinitrobenzene. It is postulated that zinc depression of contractile force is not due to metabolic inhibition, probably not due to quinidine-like action on the cell membrane, but may be due to an interference in the handling of calcium by the cell.     

Role of sex dimorphism in the pharmacological action of etimizol

It has been shown that in female rats the level of ACTH, corticosterone in the blood, relative mass of adrenals, maintenance of T3, T4 in the serum and liver was significantly higher, but the activity of liver enzyme microsomes system was lower than in males; no sex differences were observed in myocardial creatine phosphokinase system. The influence of the etimizol on the female rats significantly speed up amidopyrine N-demethylation and biotransformation of hexobarbital. In males these systems react less on etimizol, but it reduces the speed of amidopyrine N-demethylation.     

Treatment of experimental destruction of the duodenum with preparations with nootropic action

In the experiments on rats the peptic ulcer of duodenum was simulated by means of mechanic compression of reflexogenic zones of pylorus and intestine. The destruction of duodenal mucosa was developed in 3-4 hours after the compression and retained in the course of 5-6 days. Piracetam (200 mg/kg) and etimizol (3 mg/kg) eliminated and cimetidine (25 mg/kg), solcoseryl (0.5 mg/kg) and metacine (5 mg/kg) diminished the destruction after the course of 6 injections of each drug with an interval of 10-12 hours. Therapeutic effects of piracetam and etimizol correlated with their ability to return towards normal concentration of creatine phosphate.     

LIVER PARENCHYMAL CELL INJURY : I. Initial Alterations of the Cell Following Poisoning with Carbon Tetrachloride

The structure of the endoplasmic reticulum, plasma membrane, mitochondria, and Golgi apparatus of the liver parenchymal cell is strikingly altered within 1 hour following the administration of a single oral dose of carbon tetrachloride to rats. Progressive loss of glucose-6-phosphatase activity accompanies dispersal of the ergastoplasm. Electron microscopy reveals that these changes are associated with vacuolization of the cisternae of the granular endoplasmic reticulum, degranulation of its membranes, and the appearance of increased number of free ribosomes in the adjacent cytoplasmic matrix. Concomitantly, calcium enters the liver parenchymal cell and is sequestered by mitochondria. First increased at 30 minutes, calcium content is maximal at 1 hour and returns to normal at 2 hours. Although succinic and glutamic dehydrogenase activity patterns within the liver lobule are unaffected, liver cell mitochondria enlarge and some appear to fuse or assume cup-like configurations. Microvilli lining the space of Disse become irregularly indistinct and increasingly pleomorphic by 30 minutes when the plasma membrane becomes increasingly permeable to calcium. Golgi vesicles swell and discharge their granules during the period of poisoning studied. Although all the changes observed may be the result of direct interaction of carbon tetrachloride with the membranes of the cytoplasmic constituents of the liver parenchymal cell, the possibility that the irreversible changes observed in the granular endoplasmic reticulum may be due to the chemical interaction between the poison and this system is discussed.     

小麦条锈菌胞质游离钙离子动态检测方法的建立

胞质游离钙离子变化与植物病原真菌侵染寄主的动态过程具有重要的关联性。本研究以侵染小麦叶片的条锈菌31号生理小种(CYR31)为材料,以孵育法将Ca2+荧光探针Fluo-3-AM载入到小麦条锈菌细胞中,并结合激光共聚焦扫描显微技术,建立了测定侵染过程中条锈菌胞质游离Ca2+分布的试验方法。结果表明,采用10μmol/L Fluo-3-AM装载顺次进行低温4℃孵育1h,25℃孵育1h,可获得较为理想的条锈菌胞质游离Ca2+染色结果。该方法可用于检测不同侵染阶段的小麦条锈菌细胞质游离钙离子的分布变化,为进一步研究锈菌胞内钙离子动态与侵染寄主的关联性提供了技术支撑。     

Rapid flow-induced responses in endothelial cells

Endothelial cells alter their morphology, growth rate, and metabolism in response to fluid shear stress. To study rapid flow-induced responses in the 3D endothelial cell morphology and calcium distribution, coupled fluorescence microscopy with optical sectioning, digital imaging, and numerical deconvolution techniques have been utilized. Results demonstrate that within the first minutes of flow application nuclear calcium is increasing. In the same time frame whole cell height and nuclear height are reduced by about 1 microm. Whole cell height changes may facilitate reduction of shear stress gradients on the luminal surface, whereas nuclear structural changes may be important for modulating endothelial growth rate and metabolism. To study the role of the cytoskeleton in these responses, endothelial cells have been treated with specific disrupters (acrylamide, cytochalasin D, and colchicine) of each of the cytoskeleton elements (intermediate filaments, microfilaments, and microtubules, respectively). None of these compounds had any effect on the shear-induced calcium response. Cytochalasin D and acrylamide did not affect the shear-induced nuclear morphology changes. Colchicine, however, completely abrogated the response, indicating that microtubules may be implicated in force transmission from the plasma membrane to the nucleus. A pedagogical model based on tensegrity theory principles is presented that is consistent with the results on the 3D endothelial morphology.     

A dual effect of calcitonin gene-related peptide on plasma calcium levels in the chick

Calcitonin gene-related peptide (CGRP) lowers plasma calcium in the rat and inhibits bone resorption by isolated rat osteoclasts. In our preliminary studies we found that rat CGRP elevates plasma calcium levels in the chick, a response that was somewhat similar to that of parathyroid hormone. Here, we report that human CGRP (alpha) produces a concentration-dependent elevation of plasma calcium levels. The two peptides did not follow precisely the same time course. Whereas at 15 minutes CGRP produced hypocalcaemia relative to the control plasma calcium levels, at 30 minutes both CGRP and PTH were found to be hypercalcaemic. These studies suggest that CGRP initially interacts with the calcitonin receptor to produce a calcitonin-like effect, which is followed by hypercalcaemia presumably by antagonising the action of endogenous circulating calcitonin.     

Bethanechol inhibition of chicken intestinal brush border Na/H exchange: role of protein kinase C and other calcium-dependent processes.

Bethanechol, a muscarinic agonist, inhibits the initial rate of amiloride-sensitive Na uptake by intact mucosa of avian small intestine as well as by isolated chicken villus enterocytes, an effect that is maximal at 90 seconds and reverses by 6 minutes. Bethanechol similarly decreases intracellular pH in isolated cells suspended in bicarbonate-free buffer in a time course similar to inhibition of enterocyte Na uptake, suggesting inhibition of Na/H exchange. In brush border membrane vesicles rapidly prepared from cells stimulated with bethanechol, proton-dependent 22Na uptake is transiently inhibited in a time course similar to inhibition of cell Na uptake. Bethanechol also stimulates transient translocation of protein kinase C from the cytosol to the particulate fraction, a portion of this activity translocating to the brush border membrane. To determine the calcium dependence of bethanechol's action, enterocytes were loaded with varying concentrations of the calcium buffering agent quin-2. Inhibition of cell Na uptake by the calcium ionophore ionomycin could be completely reversed by quin-2 buffering in a concentration-dependent manner. In contrast, quin-2 buffering had little or no effect on the inhibition of Na uptake caused by the protein kinase C activators phorbol esters and oleoylacetylglycerol. Bethanechol's inhibitory effects were partially, but not completely reversed by quin-2 buffering. These data suggest that the effects of bethanechol on chicken villus enterocyte brush border Na/H exchange are mediated by calcium-dependent process(es) as well as by protein kinase C.     

Molecular pathways underlying the modulation of T-type calcium channels by neurotransmitters and hormones

Low-voltage-activated T-type calcium channels are expressed in various tissues, especially in the brain, where they promote neuronal firing and are involved in slow wave sleep and absence epilepsy. While the transduction pathways by which hormones and neurotransmitters modulate high-voltage-activated calcium channels are beginning to be unraveled, those implicated in T-type calcium channel regulation remain obscure. Several neurotransmitters and hormones regulate native T-type calcium channels, although some contradictory data have been reported depending on the cell type studied. This review focuses on the short-term (minutes range) modulation of T-type calcium channels by neurotransmitters and hormones and on the roles of G proteins and protein kinases in these modulatory effects. Results obtained in different native tissues are discussed and compared with the more recent studies of the three cloned T-type calcium channels CaV3.1, CaV3.2 and CaV3.3 in expression systems.     

Fertilization stimulates lipid peroxidation in the sea urchin egg

Arachidonic acid is rapidly taken-up by Strongylocentrotus purpuratus eggs and eventually incorporated into cellular lipids. During the first few minutes following fertilization the arachidonic acid that has not been incorporated into other lipid forms is oxidized to a hydroxy-fatty acid. In vivo, the time of arachidonic acid conversion coincides with the transient period of increased intracellular free calcium after fertilization. In vitro, this lipid peroxidizing activity has been shown to be initiated by micromolar calcium. Taken together with the presence of Ca2+-stimulated lipase, these results suggest that calcium regulates both the release of polyunsaturated fatty acids from cellular lipids and their subsequent oxidation. The physiological function of lipid hydroxides or hydroperoxides in sea urchin fertilization is unknown. A possibility is that they may be important in regulating the many membrane permeability changes occurring within minutes after fertilization.     

Pretreatment with Apoaequorin Protects Hippocampal CA1 Neurons from Oxygen-Glucose Deprivation

Ischemic stroke affects ∼795,000 people each year in the U.S., which results in an estimated annual cost of $73.7 billion. Calcium is pivotal in a variety of neuronal signaling cascades, however, during ischemia, excess calcium influx can trigger excitotoxic cell death. Calcium binding proteins help neurons regulate/buffer intracellular calcium levels during ischemia. Aequorin is a calcium binding protein isolated from the jellyfish Aequorea victoria, and has been used for years as a calcium indicator, but little is known about its neuroprotective properties. The present study used an in vitro rat brain slice preparation to test the hypothesis that an intra-hippocampal infusion of apoaequorin (the calcium binding component of aequorin) protects neurons from ischemic cell death. Bilaterally cannulated rats received an apoaequorin infusion in one hemisphere and vehicle control in the other. Hippocampal slices were then prepared and subjected to 5 minutes of oxygen-glucose deprivation (OGD), and cell death was assayed by trypan blue exclusion. Apoaequorin dose-dependently protected neurons from OGD – doses of 1% and 4% (but not 0.4%) significantly decreased the number of trypan blue-labeled neurons. This effect was also time dependent, lasting up to 48 hours. This time dependent effect was paralleled by changes in cytokine and chemokine expression, indicating that apoaequorin may protect neurons via a neuroimmunomodulatory mechanism. These data support the hypothesis that pretreatment with apoaequorin protects neurons against ischemic cell death, and may be an effective neurotherapeutic.     

Electroporation-induced formation of individual calcium entry sites in the cell body and processes of adherent cells.

Electroporation is a widely used method for introducing macromolecules into cells. We developed an electroporation device that requires only 1 microl of sample to load adherent cells in a 10-mm2 surface area while retaining greater than 90% cell survivability. To better understand this device, field-induced permeabilization of adherent rat basophilic leukemia and neocortical neuroblastoma cells was investigated by using fluorescent calcium and voltage indicators. Rectangular field pulses led to the formation of only a few calcium entry sites, preferentially in the hyperpolarized parts of the cell body and processes. Individual entry sites were formed at the same locations when field pulses were repeated. Before calcium entry, a partial breakdown of the membrane potential was observed in both polar regions. Based on our results, a model is proposed for the formation and closure of macromolecule entry sites in adherent cells. First, the rapid formation of a large number of small pores leads to a partial membrane potential breakdown in both polar regions of the cell. Second, over tens of milliseconds, a few entry sites for macromolecules are formed, preferentially in the hyperpolarized part of cell body and processes, at locations defined by the local membrane structure. These entry sites reseal on a time scale of 50 ms to several seconds, with residual small pores remaining open for several minutes.