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Chloride ion transport in transformed normal and cystic fibrosis epithelial cells.

The inability of beta-adrenergic agonists (eg. isoproterenol) to activate a specific apical membrane chloride channel in epithelial cells is characteristic of cystic fibrosis (CF). The study of these channels has been facilitated by the transformation of human airway epithelial cells. Clonal populations of SV40 large T-antigen transformed airway epithelial cells from both normal and cystic fibrosis individuals have been established. A limitation in the use of these cell lines has been the loss of the ability to form tight junctions after multiple subcultures. In particular, this loss appears to be associated with cell "crisis". A selection protocol that involves growing cells in medium that is high in Ca2+ and supplemented with fetal bovine serum (FBS) has been employed to facilitate progression through crisis. One cell line (1HAEo-) that has a normal phenotype and retains tight junctions post-crisis has been produced using this protocol. Immunofluorescent staining with a monoclonal antibody to the E-cadherin adhesion molecule shows a characteristic pericellular localization, indicating the presence of the junctional complex. The presence of tight junctions has been confirmed by electron microscopy. These cells produce elevated (greater than 30 fold) levels of cAMP in response to exposure to isoproterenol or forskolin. Chloride ion transport, as measured by 36Cl- efflux, is stimulated greater than 2 fold by these agents. Three post-crisis CF cell lines which grow in serum-containing medium have been established. These do not exhibit tight junctions. Elevated (greater than 25 fold) levels of cAMP are detected in these cells after addition of isoproterenol or forskolin, but this increase in cAMP is not accompanied by an increase in 36Cl-efflux. Both normal (1HAEo-) and CF cells show increased 36Cl-efflux following addition of the calcium ionophore A23187. (ABSTRACT TRUNCATED AT 250 WORDS)

Biological Transport, Active↗

Insecticidal activity of glufosinate through glutamine depletion in a caterpillar.

The herbicide glufosinate-ammonium (GLA) is a competitive inhibitor of glutamine synthetase (GS), an enzyme converting glutamate to glutamine in both plants and animals. Because GS is essential for ammonia detoxification in plants, GLA treatment disrupts photorespiration by causing a build-up of ammonia and a loss of glutamine in plant tissues. This study reports that GLA applied to leaf surfaces is also toxic to 5th-instar caterpillars of the skipper butterfly Calpodes ethlius (LD50 = 400 mg kg-1). After ingesting GLA, caterpillars stopped feeding and became dehydrated through a loss of rectal function. Caterpillars showed symptoms of neurotoxicity, such as proleg tremors, body convulsions and complete paralysis before death. Incubation of several tissues isolated from normal feeding-stage caterpillars with the GS substrates glutamate and ammonium showed that GLA inhibited GS activity in vitro. Within 24 h of ingesting GLA, caterpillars had a greatly reduced glutamine content and the ammonium ion levels had more than doubled. Injection of ammonium chloride into non-GLA-treated caterpillars had no deleterious effect, suggesting that glutamine depletion, and not a rise in body ammonium, was the primary cause of GLA toxicity following GS inhibition. This was supported by the observation that the onset of the symptoms of GLA poisoning could be postponed by giving GLA-fed caterpillars several subsequent daily injections of glutamine. The effective GLA dose fed to 5th-instar caterpillars in this study was comparable to the amount that might realistically by acquired from feeding on GLA-treated crops.

Amino Acids↗

Laser photoacoustic spectra of Sm3+ ion in Sm2O3 and SmCl3.6H2O in the spectral profile 484-542 nm.

Microphone based photoacoustic (PA) spectrometer to study solids in powder form was designed and fabricated. Laser PA spectra of Sm3+ ion in Sm2O3 and SmCl3.6H2O microcrystalline powders were recorded first time in 484-542 nm spectral region at room temperature. Analysis of these PA spectra shows new information on the Stark components of ground and excited states of Sm3+ ion. A comparison of Stark energy levels of Sm3+ ion in both oxide and hexahydrated chloride hosts is presented here.

Acoustics↗

Slight decrease in bone mineralization in cow milk-sensitive children.

BACKGROUND: Patients with cow milk allergy (CMA) are potentially at risk for osteopenia because their milk-free diet usually contains a low calcium content. In our study, different parameters of bone mineralization in children with CMA were investigated. PATIENTS AND METHODS: Twenty-seven CMA patients (mean age, 4.3 years; range, 3-8 years) were enrolled in the study. During a mean milk-free diet period of 11.8 months, children were fed extensively hydrolyzed or soy-based formulas. After a milk challenge test, 7 patients showed allergic symptoms, and the other 20 children had transient CMA. From the sera of all patients, the levels of sodium, potassium, chloride, calcium, phosphate, and magnesium ions, as well as alkaline phosphatase (AP), parathyroid hormone (PTH), osteocalcin, and beta-crosslaps, were determined. These values were compared with those of 20 healthy age-matched controls. Bone mineral density was measured as well. RESULTS: The AP and PTH concentrations were higher in CMA patients than in the control group (AP: 610.2 U/L vs. 499.7 U/L, P < 0.01; PTH: 1.56 pmol/L vs. 0.83 pmol/L, P< 0.03), but all values were in the normal range. The osteocalcin concentration was similar in both groups, and the beta-crosslaps concentration was lower in CMA patients than in controls (0.92 vs. 1.47 ng/mL, P< 0.001). The mean Z score of bone mineral density in patients with CMA was -0.6. In 10 cases, the Z score was less than the -1 SD value. On the basis of the Z score, CMA patients were divided into two groups. The PTH concentration was significantly elevated in the group with lower Z score (2.24 pmol/L vs. 1.16 pmol/L; P< 0.03). CONCLUSION: The results suggest that, in children with CMA who are on a cow milk-free diet, slight disturbances of bone mineralization can be observed by osteodensitometry.

Alkaline Phosphatase↗

Alterations in the levels of ions in blood and liver of freshwater fish, Cyprinus carpio var. communis exposed to dimethoate.

The fingerlings of Cyprinus carpio var. communis were exposed to sublethal concentration of dimethoate for 7, 14 days to evaluate the impact of the pesticide dimethoate on different ions namely sodium, potassium, chloride, calcium, magnesium. The blood potassium, calcium, magnesium and liver chloride and magnesium levels were elevated under sublethal condition. The blood sodium, chloride and liver sodium, potassium, and calcium levels were found to be significantly decreased.

Animals↗

Interaction of vanadate and chloride in chicks.

Six experiments have been conducted examining the interaction of dietary sodium chloride levels and vanadate toxicity in chicks. Increasing the dietary supplement of NaCl from 0.5 to 2.0% resulted in amelioration of vanadate toxicity, as measured by growth rate. The amelioration was found to reside in the chloride ion. Hepatic, renal, and femur vanadium concentrations were usually reduced at the higher levels of NaCl supplementation, but there was little correlation between these reductions and the reversal of vanadate toxicity.

Animals↗

Cadmium inhibits GABA-activated ion currents by increasing intracellular calcium level in snail neurons.

Blocking of the GABA-activated chloride current by cadmium was investigated in identified nerve cells (RPeD1, RPaD1) of the pond snail (Lymnaea stagnalis L.), using a two-microelectrode voltage-clamp technique. Cd2+, at 50 microM extracellular concentration, inhibited GABA-activated chloride currents, both in normal and Ca2+-free solution. Intracellular injection of Ca2+ or the application of caffeine mimicked the inhibitory effect of Cd2+ on GABA-elicited currents. Cd2+-block was eliminated, or it was substantially decreased, when neurons were intracellularly loaded with EGTA, or when the Ca2+-release was blocked by ruthenium red. The blocking effect of Cd2+ was also eliminated by applying G-protein inhibitors: pertussis toxin, suramin or GTP-gamma-S. Finally, intracellularly injected Cd2+ was ineffective at eliciting an inward current on GABA-activated currents, suggesting that the Cd2+-binding site resides extracellularly. These results suggest that cadmium inhibited GABA-activated chloride currents by increasing the intracellular Ca2+ level, by releasing it from intracellular stores and by interacting with a putative G-protein-coupled cell-surface "metal-receptor".

Animals↗

Milrinone produces a positive inotropy but is not inhibitory to gastric ion transport.

The cardiac drugs ouabain and milrinone are positive inotropic agents. Since ouabain has inhibitory effects on ion transport in the gastrointestinal tract associated with vasoconstriction and hypoxia, milrinone needed to be tested also. This report indicates that therapeutic levels of milrinone on either side of the isolated stomach wall of the guinea pig has no significant effects on active chloride ion transport or the electrical parameters of the tissue. To verify that milrinone was active in the heart at these same levels, contractility of the isolated heart of the guinea pig was measured. Milrinone significantly increased ventricular pressure and pressure development. Thus milrinone may be expected to exert its inotropic stimulation of the heart during heart failure without compromising gastrointestinal functions.

Animals↗

The different actions of chloride and potassium on postsynaptic inhibition of an isolated neurone.

Isolated stretch receptor neurones from freshwater crayfish were examined in solutions containing different concentrations of chloride and potassium. In normal solution the inhibitory reversal potential (Eipsp) of this preparation was strictly negative with respect to the resting potential and even to the reversal potential of spike after-hyperpolarization. The time courses of resting potential and Eipsp following rapid solution change suggest that the current generating the IPSP is mainly carried by chloride ions and that the participation of potassium is very small. This has also been confirmed by the calculated conductances of the activated inhibitory membrane in the different solutions. The results add further evidence for an outwardly directed pumping of chloride ions which keeps the intracellular concentration of this anion at the low level necessary for hyperpolarizing inhibition.

Animals↗

Inhibition of brain cation pump enzyme by in vitro lead ion: effects of low level [Pb] and modulation by homogenate.

Rat brain homogenates were preincubated with lead chloride for 20 min at 0 degrees C. Inhibition of K-pnitrophenylphosphatase (K-pNPPase), which measures the dephosphorylation step of Na,K-ATPase, reached steady state within 10 min and was readily reversible. The activity in brain, homogenates prepared from 60-day-old rats, expressed (mean +/- SE) as a percentage of control (100.0 +/- 0.48%), fell to 89.4 +/- 0.52% at 0.25 microM [Pb] to 64.2 +/- 1.65% at 5 microM [Pb]. However, between 5 and 25 microM [Pb] K-pNPPase activity significantly increased (to 85.4 +/- 3.14% at 25 microM) before it again fell above 30 microM. Similar results were obtained with brain homogenates prepared from 10-day-old rats. The multiphasic nature of this dose-response curve did not change with changes in buffer, substrate, anion, test tube order, or test tube composition (glass vs plastic), and appeared to be intrinsic to homogenate-Pb interactions. Microsomal preparations also exhibited multiphasicity, but with a shift in the inflection points. However, there was no multiphasicity with commercial hog brain Na,K-ATPase, suggesting an interaction between a component in the rat brain preparations and lead ion. Addition of boiled rat brain homogenate to commercial Na,K-ATPase resulted in partial protection from lead inhibition but did not produce multiphasicity over the ranges tested. Bovine serum albumin provided less protection. We conclude that there is a complex interaction among K-pNPPase activity, some factor in the rat brain homogenate, and low levels of lead ion.

4-Nitrophenylphosphatase↗

Kinetics of radiolabelled silver uptake and depuration in the gills of rainbow trout (Oncorhynchus mykiss) and European eel (Anguilla anguilla): the influence of silver speciation.

We examined the influence of speciation on the kinetics of silver uptake and depuration in the gills of two freshwater fish, the rainbow trout (Oncorhynchus mykiss) which has high branchial Na(+) and Cl(-) uptake rates and is relatively sensitive to silver, and the European eel (Anguilla anguilla, yellow stage) which has low ion uptake rates and is relatively resistant to silver. Fish previously acclimated to the appropriate chloride level were exposed to 110mAgNO(3) (1.3 microg l(-1), sublethal) for 24 h in synthetic softwater with either low (10 microM) or high (1200 microM) chloride concentration, and then followed over a subsequent 67-day post-exposure period in silver-free water of the same chloride content. The exposures were therefore mainly to the free ion, Ag(+) in the low chloride water versus mainly to the neutral aqueous complex, AgCl(aq) in the high chloride water. In trout, but not in eel, water chloride is known to protect against physiological disturbances and toxicity caused by Ag(+). In both fish species, at both chloride levels, silver uptake exhibited complex kinetics. Gill silver loading occurred slowly until 6 h, then rose greatly to a peak at 12 h, followed by significant net depuration thereafter during continued exposure. By 24 h, net gill loading was three- to fivefold greater from AgCl(aq) than from Ag(+) exposure in both species, and threefold greater in trout than in eel under both conditions, with trout holding a lower fraction of the whole body burden in their gills. During the post-exposure period, depuration of silver from the gills occurred rapidly in trout, but very slowly in eel, such that gill silver burdens were greater in eel throughout the 67-day period on both an absolute and relative basis (e.g. 35% of whole body burden in eel versus <3% in trout at day 8). The kinetics of depuration were described by two phase exponential models, with break points between the fast and slow phases at 1 and 15 days for trout and eel, respectively. We conclude that speciation affects not only uptake rates but also the kinetics of depuration. When silver is loaded from AgCl(aq) it is clearly more labile than from Ag(+) exposures, with 1.6-1.8-fold greater loss rates during the fast phases in both species. Differences in branchial silver uptake between eel and trout correlate well with differences in acute toxicity, but are not as large as differences in ion uptake rates. The complex time-dependent patterns of gill loading, and the higher loading from AgCl(aq) than from Ag(+), mean that gill total silver burden is not an appropriate endpoint for biotic ligand modelling.

Adaptation, Physiological↗

Alterations in gastric mucus secretion in rhesus monkeys after exposure to ionizing radiation.

The aim of the present study was to evaluate the effect of gamma-irradiation on soluble gastric mucus. Six conscious chair-adapted rhesus monkeys were studied once before and twice after exposure to ionizing irradiation (800 rads). Using a marker (99mTc-DTPA) dilution technique, acidic glycoprotein (AG), neutral glycoprotein (NG), ion, and fluid output were determined during a basal period and after the administration of an 80-ml water load. Irradiation significantly increased the outputs of both AG and NG during the basal period. After the water load, NG output remained elevated but irradiation abolished postload AG output thus inhibiting the normal rise in AG output stimulated by the load. Two days after irradiation NG output had returned to control levels whereas AG output was still suppressed. Sodium and potassium ion outputs were unaltered by irradiation. Chloride and fluid outputs were significantly inhibited on the day of irradiation but had returned to control levels within 3 days. These results indicate that irradiation produces significant changes in both the quantity and nature of the soluble mucus glycoproteins secreted into the gastric juice. It is suggested that these changes may compromise the protective ability of gastric mucus.

Animals↗

Dissociation dynamics of propargyl chloride molecular ion near the reaction threshold: manifestation of quantum mechanical tunneling

The Cl loss from the propargyl chloride molecular ion has been investigated using mass-analyzed ion kinetic energy spectrometry (MIKES). The kinetic energy release distribution in the unimolecular dissociation has been determined. The potential energy surface for the mechanistic pathway has been calculated at the B3LYP/6-311G** density functional theory level. The calculated potential energy surface suggested that the threshold dissociation of the propargyl chloride molecular ion produces the C(3)H(3)(+) ion, only with the cyclopropenium structure, and with the release of a large amount of kinetic energy. This is in agreement with experimental results. Also, calculation of the rate constants with statistical rate models predicted that the reaction observed on a microsecond time scale occurs via tunneling through the rate-determining isomerization barrier for H-atom transfer. It has been found that a broad lifetime distribution is a manifestation of quantum mechanical tunneling of a precursor prepared under thermal conditions. Reinterpretation of previous photoelectron-photoion coincidence results taking into account the tunneling effect necessitated raising the critical energy to 0.64 eV from the energy of 0.34 eV reported previously. Copyright 2000 John Wiley & Sons, Ltd.

Journal Article↗

Chlormethiazole--mode of action.

Studies in mice demonstrated that the anticonvulsant profile of chlormethiazole differs from that of diazepam and the barbiturates. Chlormethiazole protects animals from convulsions induced by a wide variety of chemoconvulsants known to block the action of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA), such as bicuculline, picrotoxin, isoniazid and pentetrazol, thus confirming and extending earlier studies on its broad anticonvulsant characteristics. Chlormethiazole is particularly potent against isoniazid-induced convulsions, which are probably induced by reductions of GABA levels in the brain. Chlormethiazole was found to have a weak action on benzodiazepine receptor binding, GABA receptor binding and kainic acid receptor binding. Chlormethiazole inhibited picrotoxin binding at very high concentrations, but lowered the functional effects of picrotoxin at much lower concentrations than those affecting picrotoxin binding. Moreover, chlormethiazole failed to change GABA or glutamate levels in the brain and did not affect glutamic acid decarboxylase (GAD) activities in the rat brain. Muscimol (a GABAA agonist) enhanced the anticonvulsant activity of chlormethiazole against picrotoxin but not against bicuculline-induced convulsions. Muscimol enhanced the anticonvulsant potency of diazepam against both chemoconvulsants. These data suggest that the anticonvulsant activity of chlormethiazole is not mediated directly through changes in GABA or glutamate levels or by a direct (agonist) action at the GABA or benzodiazepine receptor complex. These findings suggest that chlormethiazole may enhance GABA transmission beyond the GABA receptors, hypothetically at the level of the GABA receptor coupled ionophore (e.g. the chloride ion channel). Applied micro-iontophoretically, chlormethiazole was found to potentiate the inhibitory responses to GABA, muscimol and glycine, but not to acetylcholine. The potentiation of glycine-mediated inhibition is unique for chlormethiazole and does not occur with any other known anticonvulsant (barbiturates, benzodiazepine, phenytoin or sodium valproate). Studies in primary cultures, derived from spinal cord neurones, showed that chlormethiazole produces hyperpolarization together with an increase in the threshold for action potential generation. Further in vitro studies indicated that chlormethiazole acts on some types of Ca2+-dependent chloride ion channels.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Soil chemistries define aquatic habitats with immature populations of the Culicoides variipennis complex (Diptera: Ceratopogonidae).

Aquatic habitats with immature populations of the Culicoides variipennis (Coquillett) complex were sampled across the United States and tested for soil chemistry parameters. Indicators of organic loading, phosphate, percentage of organic matter, and nitrate were high for C. variipennis and C. sonorensis, reflecting the common occurrence of larvae in sediments contaminated with livestock waste; these factors were low in western playa lake habitats with immature C. occidentalis. Levels of salt-forming ions and indicators of salinity, notably sodium, chloride, magnesium, sulfate, boron, and conductivity, were elevated in substrates supporting all 3 species, with values that declined from very high to high across the arid west to moderate east of the Mississippi river, and differed significantly among species. Concentrations of boron differed most strongly among habitats of the respective species, ranging from exceptionally high values in playa lake substrates with C. occidentalis, to high and moderate levels in western and southeastern sites with C. sonorensis, to moderate levels for substrates in the east with C. variipennis. The presence of C. sonorensis in dairy wastewater lagoons and saline springs in geographic regions that are otherwise inhabited by C. variipennis may reflect salinity factors that regulate the distribution of the C. variipennis complex.

Animals↗

The role of high ionic concentrations in protection against X-irradiation.

Various levels of protection against x-irradiation damage in bacteriophage T1 may be obtained by the addition of inorganic salts to the aqueous virus suspensions during irradiation. The highest survival values are obtained with the nitrite salts, and their protective power is attributed primarily to their function as reducing agents. The nitrate ion shows greater protection than the corresponding sulfate or chloride ions. This may be due in part to the lower energy level of the nitrate ion, by reason of resonance. Since greater expenditure of incident energy is required to raise the ion from the ground state, the energy thus dissipated may be ineffective in the inactivation of virus particles. The ammonium salts exhibit protection of a different order of magnitude from that of the metallic salts. It is postulated that NH(4) (+) protects in a threefold way: (a) dehydration, (b) reduction, in which the ammonia is oxidized to nitrite and the nitrite to nitrate, and (c) stabilization of the virus protein. Metallic salts likewise protect, but a point of maximum protection is reached in lower concentrations than in the case of the ammonium salts. After this maximum protection is reached, there is a rapid decline in survival with increased concentration. This prevents protection of the order of magnitude that can be obtained with the ammonium salts. It is postulated that a specific cationic interaction with the phage may be responsible for the decreased protection. Bacteriophage is protected during x-irradiation by an alkaline pH, in the case of NH(4)OH. This protection could not be produced with NaOH, presumably because of the greater hydrolysis of the protein components of the virus particle in solutions of NaOH, whereas NH(4)OH stabilizes the protein.

Bacteriophages↗

GABA-gated chloride ion influx in brains of epileptic El mice.

GABA-gated chloride ion influx was measured in brain "microsac" preparations of epileptic El mice. There was significantly greater sensitivity to GABA in stimulated El mice (which had 14-18 convulsions induced at weekly intervals) than in unstimulated El mice (which had not experienced convulsions) or ddY mice. GABA-gated chloride ion influx was significantly decreased 20 min after a single convulsion, and returned to the preconvulsion level 60 min after a convulsion. These findings suggest that the functional state of GABA-gated chloride channel in El mice is changed secondarily by single or repeated convulsions.

Animals↗