Evaluation of Cuprimine® and Syprine® for decorporation of radioisotopes of cesium, cobalt, iridium and strontium.

Cuprimine® and Syprine® are therapeutics approved by the USFDA to treat copper overload in Wilson Disease (a genetic defect in copper transport) by chelation and accelerated excretion of internally-deposited copper. These oral therapeutics are based on the respective active ingredients D-penicillamine (DPA) and N,N'-bis (2-aminoethyl) -1,2-ethanediamine dihydrochloride (Trien). Cuprimine is considered the primary treatment, although physicians are increasingly turning to Syprine as a first-line therapy. Both drugs exhibit oral systemic activity and low toxicity; their biological effects and safety are established. Previous in vivo studies using a rodent animal model established the decorporation potential of Cuprimine and Syprine for (60)Co and (210)Po. Currently these studies are being expanded to evaluate the in vivo decorporation efficacy of these drugs for several additional radionuclides. In this report, results of this investigation are discussed using the radionuclides (137)Cs, (60)Co, (192)Ir and (85)Sr. Short-term 48-h pilot studies were undertaken to evaluate DPA and Trien for their in vivo decorporation potential using male Wistar-Han rats. In these studies, a radionuclide solution was administered to the animals by intravenous (IV) injection, followed by a single IV dose of either DPA or Trien. Control animals received the radionuclide alone. Results show effective decorporation of (60)Co by DPA within the time frame evaluated. DPA and Trien were also modestly effective in decorporation of (137)Cs and (85)Sr, respectively. The study did not find DPA or Trien effective for decorporation of (192)Ir. Based on these encouraging findings, further studies to evaluate the dose-response profiles and timing of the chelator administration post exposure to radionuclides are warranted.

Characteristics of cesium accumulation in the filamentous soil bacterium Streptomyces sp. K202.

A filamentous soil bacterium, strain K202, was isolated from soil where an edible mushroom (Boletopsis leucomelas) was growing and identified as belonging to the genus Streptomyces on the basis of its morphological characteristics and the presence of LL-2, 6-diaminopimelic acid. We studied the existence states of Cs and its migration from extracellular to intracellular fluid in the mycelia of Streptomyces sp. K202. The results indicated that Cs accumulated in the cells through at least 2 steps: in the first step, Cs(+) was immediately and non-specifically adsorbed on the negatively charged cell surface, and in the second step, this adsorbed Cs(+) was taken up into the cytoplasm, and a part of the Cs entering the cytoplasm was taken up by an energy-dependent transport system(s). Further, we confirmed that a part of the Cs(+) was taken up into the mycelia competitively with K(+), because K(+) uptake into the intact mycelia of the strain was significantly inhibited by the presence of Cs(+) in the culture media. This suggested that part of the Cs is transported by the potassium transport system. Moreover, (133)Cs-NMR spectra and SEM-EDX spectra of the mycelia that accumulated Cs showed the presence of at least 2 intracellular Cs states: Cs(+) trapped by intercellular materials such as polyphosphate and Cs(+) present in a cytoplasmic pool.

Development of functional I f channels in mMSCs after transfection with mHCN4: effects on cell morphology and mechanical activity in vitro.

OBJECTIVE: To study the functional properties of I(f) channels and the changes in mechanical activity of mouse mesenchymal stem cells (mMSCs) transfected with mHCN4. METHODS: mMSCs were purified by using CD11b-immunomagnetic microbeads and transfected with pMSCV-mHCN4-EGFP or pMSCV-EGFP. We examined the kinetic characteristics of the mHCN4 channel. The morphological changes of positively transfected mMSCs were investigated at the same time. RESULTS: The I(f) current recorded from the experimental group was sensitive to extracellular Cs(+) (-44.5 +/- 4.2 vs. -5.5 +/- 1.0 pA/pF, p < 0.001). The half-maximal activation was -99.0 +/- 5.8 mV. The time constant of activation was 451 +/- 61 ms under -140 mV. The control cells did not show the current under the same conditions. The absolute values of half-maximal activation decreased in the presence of cAMP or cGMP in the experimental group (-78.6 +/- 10.4 and -85.7 +/- 8.6 vs. -99.0 +/- 5.8 mV, respectively, p < 0.05). mMSCs transfected with pMSCV-mHCN4-EGFP could form spontaneous beating cells. Extracellular Cs(+) decreased the beating rate significantly (196 +/- 50 vs. 66 +/- 23 bmp, p < 0.01). CONCLUSIONS: Functional I(f) channels can be reconstructed in mMSCs infected with mHCN4. mMSCs modified by successful transfection with mHCN4 can differentiate so as to develop spontaneous mechanical activity.

The toxic function of cesium 5-sulfosalicylate based on the investigation of its trans-erythrocytes membrane behaviors and morphological properties.

In order to evaluate the cesium-induced toxic functional changes in organisms, transmembrane activities of cesium 5-sulfosalicylate (Cs(H(2)Ssal)) into human erythrocyte in vitro is presented in this paper, including kinetic characteristic of transport process and pathways involved in it. The uptake amount of Cs(H(2)Ssal) by erythrocyte was determined both by Graphite Furnace Atomic Absorption Spectrometry (GFAAS) and spectrofluorimetry. The pathways of Cs(H(2)Ssal) transporting into erythrocyte are proposed according to inhibition investigation. The influence of Cs(H(2)Ssal) on morphological properties of erythrocytes was examined using Scanning Electron Microscopy (SEM) to determined the endurable concentration extent of erythrocytes to Cs(H(2)Ssal). Results show that transmembrane of Cs(H(2)Ssal) has characteristic of first-order kinetic process during the first 2h, and four pathways were involved in its transporting activities: Ca(2+) channel, Na(+)-K(+) pump, Na(+)-Cs(+) countertransport, and anion Cl(-)/CsCO(3)(-) exchange. The transmembrane process of Cs(H(2)Ssal) can both prevent the uptake of K(+) and induces abnormal accumulation of extracellular K(+) as well as occupy some K(+)-binding sites in protein, causing some tissues losing their activities and functions. Only high concentrations of Cs(H(2)Ssal) could change morphological properties of erythrocytes greatly and cause hemolysis eventually.

Comparison on plasma caesium kinetics in goats and horses with special emphasis on exercising horses.

AIMS: Like potassium (K+), caesium (Cs+) tends to concentrate intracellularly. The aim here was to determine how moderate exercise affects the uptake of Cs+ from blood plasma. METHODS: After an intravenous Cs+ dose of 5 micromol kg(-1), plasma Cs+ concentration was followed for 100 min in goats and for 60 min in horses. The latter were divided into two groups, one resting and the other trotting on a treadmill (inclination 3 degrees, speed 5 m s(-1)). RESULTS: The plasma Cs+ concentration follows a multiphase exponential decay curve, which initially could be approximated with a two-phase curve. The initially high rate constant (approximately 10 h(-1)) decreased to around 1 h(-1) in 40 min. Exercise more than doubled the rate of removal of Cs+ from plasma between 20 and 40 min after the start of exercise. After exercise, the rate returned to resting levels within 10 min. Plasma K+, on the contrary, declined for at least 20 min after exercise had ended. CONCLUSIONS: Moderate exercise significantly increases the rate of removal of Cs+ from the bloodstream. After exercise, the rate returns to the resting levels within 10 min. The increased rate of Cs+ removal during exercise is likely due to increased activity of Na+, K+-ATPase in working skeletal muscles.

pH modulation using CsCl enhances therapeutic effects of vitamin D in LNCaP tumor bearing mice.

BACKGROUND: The downstream effects of pH modulation significantly impact the biological fate of chemotherapeutic agents and tumor responsiveness to therapy. We have studied the effects of cesium chloride (CsCl) on pH modulation and subsequent vitamin D treatment in vitamin D receptor (VDR) positive LNCaP tumors and VDR null MDA/LCC6 tumors in vivo. METHODS: Mice bearing LNCaP or MDA/LCC6 tumors were dosed orally with CsCl (150 mg/kg) or vitamin D (1 microg/kg) alone and in combination. Tumor volume and serum PSA (LNCaP only) were measured and intracellular pH was determined, using magnetic resonance spectroscopy (MRS), at tumor and leg muscle sites. Atomic absorption spectroscopy (AAS) was used to quantitate cesium in serum, organs, and tumor tissues. RESULTS: From day 10 onwards, statistically significant (P<0.01) differences were observed in all LNCaP treated groups as compared with control. CsCl co-administered with vitamin D, caused an apparent sensitization of efficacy in this tumor model. There were no correlating differences in serum PSA. Elevation of pH was statistically significant for all three treatment groups as compared with control. The pH measured in leg muscle was not influenced by CsCl treatment. Inhibition of tumor growth was not apparent in VDR null MDA/LCC6 tumors although intracellular tumor pH was shifted. Cesium was rapidly absorbed into serum and present in LNCaP tumors, prostates and other tissues after 1 hr, remaining for up to 24 hr. CONCLUSIONS: The data presented in this manuscript is the first report of chemosensitization by in vivo pH modulation using CsCl in mice bearing prostate or any other tumor xenograft.

Contents of cesium, iodine, strontium, thorium, and uranium in selected human organs of adult asian population.

Contents of cesium, iodine, strontium, thorium, and uranium in some selected human organs were estimated for adult Asian population using data obtained in four Asian countries: China, India, Philippines, and Republic of Korea, as part of a Coordinated Research Program of the International Atomic Energy Agency on "Ingestion and Organ contents of elements of importance in radiation protection." These countries together represent more than 40% of the world population. Highly sensitive analytical techniques were employed to measure cesium in skeletal muscle, iodine in thyroid, strontium in skeleton, thorium and uranium in skeleton, liver, kidneys, and lungs where, in comparison to other organs, these elements are present in higher concentrations. The organ contents for adult Asian population, when compared with the corresponding data proposed for Reference Man by International Commission on Radiological Protection (ICRP), showed about 40 times lower kidneys content and about 10 times lower skeleton content of uranium. The content of thorium in skeleton for Asian population was also half of the ICRP Reference Man value. Interestingly, organ contents for the other elements such as iodine in thyroid, cesium in skeletal muscle, and strontium in skeleton were comparable for Asian and the Caucasian population (represented by ICRP Reference Man). Organ contents for these elements were also calculated by applying the new ICRP models of these elements to their daily intakes. The comparison of the calculated and measured organ contents showed that despite uncertainties in the organ content values arising due to the inter-country variations in daily dietary intakes, the contents were within a factor of two to three. This observation is significant since human data both on organ contents and ingestion were obtained at environmental level of intakes. The study suggests that currently available ICRP models for these elements are quite realistic.

Reduction of crop contamination by soil resuspension within the 30-km zone of the Chernobyl nuclear power plant.

A field experiment was conducted within the 30-km zone of the Chernobyl Nuclear Power Plant to analyze whether the application of mulching reduced resuspension of 137Cs contaminated soil in oat (Avena sativa) crops. In 1993, we applied a mulch treatment at a dose of 200 g m(-2), and soil resuspension was measured by estimating soil loadings onto plant surfaces from Ti concentrations in plants. In 1994, two mulch doses were applied, 200 and 50 g m(-2), and we estimated the contribution of soil resuspension by using artificial resuspension collection devices (ARC). In the 1993 experiment between 4.6 and 34.4% of the plant's total 137Cs contamination was attributed to external soil contamination. The mean amount of soil-derived 137Cs attached to vegetation was 124.7 Bq kg(-1)(plant) in control plots and 53.7 Bq kg(-1)(plant) in mulched plots. In the 1994 experiment, covering the soil with a mulch layer decreased the radiocesium content in ARC by about 70%. Results obtained in these experiments suggest that soil resuspension was a significant mechanism for plant contamination and that mulching was effective in reducing that contamination.

Blood and tissue concentration of cesium after exposure to cesium chloride: a report of two cases.

CONTEXT: Complementary alternative medicine therapies based on the use of cesium chloride preparations for the treatment of cancer and radiation poisoning, have generated therapeutic interest; but oral or intravenous administration of cesium chloride (CsCl) to cancer patients as an alternative mode of cancer therapy have not been approved by the US Food and Drug Administration (FDA). OBJECTIVE: Cesium (Cs) levels from human tissue were measured to determine exposure to an alternative medical treatment. Cesium levels are reported from two patients who were administered cesium chloride in conjunction with aloe vera as part of an alternative cancer treatment. DESIGN: The samples were analyzed by graphite furnace atomic absorption spectrometry with Zeeman background correction. As a reference, Cs was also determined in brain, liver, kidney, and whole blood from control case materials retrieved from the National Tissue Repository of the Armed Forces Institute of Pathology. RESULTS: High levels of cesium were found in brain, liver, kidney, bile, gastric content, and whole blood collected at autopsy as compared to reference levels. The administration of cesium chloride resulted in blood levels a factor of 1100 higher than normal. The highest Cs concentrations were found in the liver (1029 microg/g, dry wt), followed by the kidney (815 microg/g, dry wt) and brain (219 microg/g, dry wt). CONCLUSION: The high accumulation in the liver suggests that hepatotoxicity from Cs might be an initial presenting symptom in Cs-poisoning cases. This is the first report describing two cases with high Cs levels in human tissues.

Dependence of Na+-K+ pump current-voltage relationship on intracellular Na+, K+, and Cs+ in rabbit cardiac myocytes.

To examine effects of cytosolic Na+, K+, and Cs+ on the voltage dependence of the Na+-K+ pump, we measured Na+-K+ pump current (Ip) of ventricular myocytes voltage-clamped at potentials (Vm) from 100 to +60 mV. Superfusates were designed to eliminate voltage dependence at extracellular pump sites. The cytosolic compartment of myocytes was perfused with patch pipette solutions with a Na+ concentration ([Na]pip) of 80 mM and a K+ concentration from 0 to 80 mM or with solutions containing Na+ in concentrations from 0.1 to 100 mM and K+ in a concentration of either 0 or 80 mM. When [Na]pip was 80 mM, K+ in pipette solutions had a voltage-dependent inhibitory effect on Ip and induced a negative slope of the Ip-Vm relationship. Cs+ in pipette solutions had an effect on Ip qualitatively similar to that of K+. Increases in Ip with increases in [Na]pip were voltage dependent. The dielectric coefficient derived from [Na]pip-Ip relationships at the different test potentials was 0.15 when pipette solutions included 80 mM K+ and 0.06 when pipette solutions were K+ free.

Cesium adsorption in hydrated iron oxide particles suspensions: an NMR study.

137Cs is an important component of nuclear waste which may pollute water. Its migration in natural environments is slowed down by adsorption on minerals. Cesium adsorption on akaganeite (beta-FeOOH) particles, dextran-coated ferrihydrite (5 Fe(2)O(3)-9H(2)O) particles, and ferritin in aqueous solutions is studied with (133)Cs nuclear magnetic resonance measurements. The longitudinal relaxation time (T(1)) of (133)Cs in the presence of such magnetic particles depends on whether the ions bind to the particle or not. T(1) of (133)Cs ions in aqueous solutions containing the same amount of magnetized particles will not depend on cesium concentration if relaxation is governed by diffusion (when cesium is not able to bind), but it will depend on cesium concentration if exchange governs relaxation (when cesium is able to bind). The method is successfully tested using TEMPO, a nitroxide stable free radical whose relaxation is due to diffusion. (133)Cs relaxation in solutions of ferritin, akaganeite, and dextran-coated ferrihydrite particles is found to result from a cationic exchange of cesium ions between particles surface and bulk ions, owing to adsorption. The effect of pH on (133)Cs relaxation in solutions of the particles is consistent with the adsorption properties of cations on hydrated iron oxides.

Monovalent cation permeability and Ca(2+) block of the store-operated Ca(2+) current I(CRAC )in rat basophilic leukemia cells.

Like voltage-operated Ca(2+) channels, store-operated CRAC channels become permeable to monovalent cations in the absence of external divalent cations. Using the whole-cell patch-clamp technique, we have characterized the permeation and selectivity properties of store-operated channels in the rat basophilic leukemia (RBL-1) cell line. Store depletion by dialysis with InsP(3) and 10 mM EGTA resulted in the rapid development of large inward currents in Na(+)- and Li(+)-based divalent-free solutions. Cs(+) permeated the channels poorly (P(Cs)/ P(Na)=0.01). Trimethylamine (TMA(+)), tetramethylammonium (TeMA(+)), tetraethylammonium (TEA(+)), N-methyl- D-glucamine (NMDG(+)) and TRIS(+) were not measurably permeant. NH(4)(+) was conducted well. We estimated the minimum pore diameter under divalent-free conditions to be between 0.32 nm and 0.55 nm. When cells were dialysed with buffered Ca(2+) solution and I(CRAC) activated by application of thapsigargin, P(Cs)/ P(Na) was still low (0.08). Outward currents through CRAC channels were carried by intracellular Na(+), K(+) and, to a much lesser extent, by Cs(+). Currents were unaffected by dialysis with Mg(2+)-free solution. The Na(+) current was inhibited by external Ca(2+) (half-maximal blocking concentration of 10 microM). This Ca(2+)-dependent block could be alleviated by hyperpolarization. The monovalent Na(+) current was voltage dependent, increasing as the holding potential depolarized above 0 mV. Our results suggest that CRAC channels in RBL-1 cells have a smaller pore diameter than voltage-operated Ca(2+) channels, discriminate between Group I cations, and differ markedly in their selectivity from CRAC channels reported in lymphocytes.

Ionic selectivity of native ATP-activated (P2X) receptor channels in dissociated neurones from rat parasympathetic ganglia.

1. The relative permeability of the native P2X receptor channel to monovalent and divalent inorganic and organic cations was determined from reversal potential measurements of ATP-evoked currents in parasympathetic neurones dissociated from rat submandibular ganglia using the dialysed whole-cell patch clamp technique. 2. The P2X receptor-channel exhibited weak selectivity among the alkali metals with a selectivity sequence of Na(+) > Li(+) > Cs(+) > Rb(+) > K(+), and permeability ratios relative to Cs(+) (P(X)/P(Cs)) ranging from 1.11 to 0.86. 3. The selectivity for the divalent alkaline earth cations was also weak with the sequence Ca(2+) > Sr(2+) > Ba(2+) > Mn(2+) > Mg(2+). ATP-evoked currents were strongly inhibited when the extracellular divalent cation concentration was increased. 4. The calculated permeability ratios of different ammonium cations are higher than those of the alkali metal cations. The permeability sequence obtained for the saturated organic cations is inversely correlated with the size of the cation. The unsaturated organic cations have a higher permeability than that predicted by molecular size. 5. Acidification to pH 6.2 increased the ATP-induced current amplitude twofold, whereas alkalization to 8.2 and 9.2 markedly reduced current amplitude. Cell dialysis with either anti-P2X(2) and/or anti-P2X(4) but not anti-P2X(1) antibodies attenuated the ATP-evoked current amplitude. Taken together, these data are consistent with homomeric and/or heteromeric P2X(2) and P2X(4) receptor subtypes expressed in rat submandibular neurones. 6. The permeability ratios for the series of monovalent organic cations, with the exception of unsaturated cations, were approximately related to the ionic size. The relative permeabilities of the monovalent inoganic and organic cations tested are similar to those reported previously for cloned rat P2X(2) receptors expressed in mammalian cells.

Three types of K(+) currents in murine osteocyte-like cells (MLO-Y4).

Osteocytes play an important role in signaling within bone. Communication of osteocytes with each other and with bone lining cells may have a function in mineral homeostasis and mechanotransduction. However, very little is known of the expression of ion channels in these cells. Using the whole-cell patch-clamp technique, we have detected three types of K(+) currents in the mouse osteocyte-like cell line MLO-Y4. The most commonly observed current (48% of cells) activated rapidly (20 msec) in response to depolarizing steps from -40 mV and exhibited voltage-dependent inactivation. The current was inhibited by 20 mmol/L tetraethyl ammonium (TEA) and abolished by intracellular 2 mmol/L 4-aminopyridine (4-AP). Biophysical and pharmacological characteristics of the current differed from those of inactivating K(+) currents in osteoblastic cells. In 22% of cells, a slowly activating, voltage-activated current was observed (threshold at 20-30 mV). This current was TEA insensitive, was abolished by intracellular application of 2 mmol/L 4-AP, and was strongly inhibited by apamin, a selective inhibitor of small conductance (SK) Ca(2+)-activated K(+) channels. A third current developed during whole-cell dialysis (37% of cells). This current showed little voltage sensitivity. It was abolished by intracellular application of 2 mmol/L 4-AP, high-extracellular Ba(2+) (108 mmol/L), or by inclusion of ATP in the intracellular solution, but was insensitive to TEA, apamin, Cs(+), and glibenclamide. None of these currents was affected by replacement of chloride with acetate in the bath or pipette salines. Reverse-transcription polymerase chain reaction confirmed the presence of mRNA for the types 1 and 2 SK channels, but message for the large conductance (BK) Ca(2+)-activated K(+) channel was not detected in these cells. Message for the sulphonylurea receptor SUR2, a subunit of glibenclamide-insensitive ATP-dependent K(+) channels (K(ATP)), was also detected, but the glibenclamide-sensitive SUR1 subunit was not. These data are the first descriptions of SK- and ATP-sensitive, glibenclamide-insensitive channels in cells of osteoblastic lineage. Our findings are consistent with a change in K(+) channel expression during differentiation from osteoblast to osteocyte. K(+) channels of osteocytes will contribute to maintenance of the cell membrane potential and thus may participate in mechanosensitivity and osteocyte intercellular communication. In addition, they may be involved in homeostatic maintenance of the extracellular fluid occupying the periosteocytic space.

Reduced glutathione inhibits beta-NAD+-activated non-selective cation currents in the CRI-G1 rat insulin-secreting cell line.

1. Whole-cell voltage-clamp recordings were used to study the characteristics of a non-selective cation current, activated by intracellular beta-NAD+, present in CRI-G1 insulin-secreting cells. The monovalent cations Na+, K+ and Cs+ were equally permeant through this channel. 2. The magnitude of the beta-NAD+ current was dependent on the concentration of both beta-NAD+ and Ca2+ in the cell. The properties of the beta-NAD+-activated macroscopic current are similar to those of the beta-NAD+-activated non-selective cation channel (NSNAD) examined at the single channel level in this cell line. 3. The presence of intracellular reduced glutathione (GSH) inhibited the beta-NAD+-activated macroscopic current and the activity of the NSNAD channel in inside-out patch recordings. 4. The inhibition of beta-NAD+-activated currents by GSH is mimicked by its analogue ophthalmic acid but not by another thiol reducing agent dithiothreitol, indicating the presence of a specific GSH binding site present on the NSNAD channel or associated protein.

Transfer across the human gut of environmental plutonium, americium, cobalt, caesium and technetium: studies with cockles (Cerastoderma edule) from the Irish Sea.

Our previous studies have indicated lower values of the gut transfer factor ('f1 values') for plutonium and americium in winkles (Littorina littorea) than adopted by ICRP. The present study was undertaken primarily to investigate whether this observation extends to other species. Samples of cockles (Carastoderma edule) from Ravenglass, Cumbria were eaten by volunteers who provided 24 h samples of urine and faeces. Urine samples indicated f1 values for cockles which were higher than for winkles; for plutonium these ranged overall up to 7 x 10(-4) with an arithmetic mean in the range (2-3) x 10(-4), and for americium up to 2.6 x 10(-4) with an arithmetic mean of 1.2 x 10(-4). Limited data based on volunteers eating cockles from the Solway suggest that f1 values for americium may be greater at distance from Sellafield. The measured values compare with 5 x 10(-4) used by the ICRP for environmental forms of both elements, which would appear to provide adequate protection when calculating doses from Cumbrian cockles. Data for other nuclides were obtained by analysing faecal samples from the volunteers who ate the Ravenglass cockles. Cobalt-60 showed an f1 value in the region of 0.2, twice the value currently used by ICRP. For 137Cs, variabilities were indicated in the range 0.08 to 0.43, within the ICRP value of f1 = 1.0. Technetium-99 gave f1 values up to about 0.6, in reasonable conformity with the ICRP value of 0.5.

Chromosome aberrations in Syrian hamsters following very low radiation doses in vivo.

This paper addresses a report of a large increase (approximately 6- to 11-fold) in chromosome aberrations in lymphocytes of persons in Salzburg attributed to their exposure to fallout from the Chernobyl cloud. Their additional exposure, approximately 0.3 mGy in 1 year, comprised about a 30% increase in their normal background radiation dose. The report has attracted considerable attention because, if correct, it seriously challenges assumptions of linearity in the low-dose response for chromosomal damage and, by implication, the linear, no-threshold hypothesis for risk of induced cancer. An experiment has been carried out with Syrian hamsters treated with caesium-137 to produce a range of doses comparable with those calculated for the persons in Salzburg. No significant elevation in lymphocyte aberration yields was found in the hamsters, thus arguing against the conclusions of the Salzburg study.

Quantification of ion transport in perfused rat heart: 133Cs+ as an NMR active K+ analog.

Proper ion balance between intra- and extracellular compartments is necessary for normal physiological function. Conversely, alterations in membrane ion transport occur in numerous pathological states. As a noninvasive, nondestructive spectroscopic technique, nuclear magnetic resonance (NMR) offers a powerful approach to the study of ion balance in intact biological systems. Unfortunately, rare NMR active nuclides that are isotopes of the 100% naturally abundant 23Na+ and 39K+ are not available for tracer kinetic studies of Na1 and K+ transport. However, Cs is a biologically active analog of K+, and the 100% naturally abundant NMR active 133Cs+ nuclide can be employed to examine K+ transport (Davis, D. G., E. Murphy, and R. E. London. Biochemistry 27: 3547-3551, 1988). The distinguishing feature of 133Cs+ is that it naturally gives two separate well-resolved NMR resonances for intra- and extra-cellular 133Cs+, permitting study of the time course changes of either of these compartments independent of the other. In this report, the experimental procedures and compartmental modeling formalism are developed that allow quantitative analysis of Cs+ membrane transport in the perfused rat heart. Intracellular 133Cs+ is shown to be 100% visible by solution-state NMR methods and its influx transport to be markedly inhibited by ouabain, a confirmation of findings previously reported by others. Intracellular 133Cs+ spin-lattice and spin-spin relaxation times at 7 T were determined to be 2.1 +/- 0.3 (SD)s (n = 8) and 0.065 +/- 0.007 (SD) s (n = 8), respectively, for T1 and T2. The rate constant for Na(+)-K(+)-ATPase pump dominated intracellular influx was measured to be 0.25 +/- 0.07 (SD) min-1 (n = 27) and that for efflux 0.005 +/- 0.001 (SD) min-1 (n = 14). The rate constant for 133Cs+ equilibration in the extracellular space at supraphysiological perfusate flow rate (20 ml/min) was found to be 4.6 +/- 0.9 (SD) min-1 (n = 20). Thus extracellular diffusion limitations do not dominate the 133Cs+ transport measurements.

Inhibition of ion transport in septic rat heart: 133Cs+ as an NMR active K+ analog.

Sepsis, the systemic response to severe infection, and the resulting multiorgan failure it induces are major contributors to intensive care unit morbidity and mortality. A number of abnormalities in ion transport processes and intracellular free Na+ ([Na+]i) and K+ ([K+]i) concentrations have been reported to occur during sepsis/endotoxemia. An effect of sepsis on the NA(+)-K(+)-ATPase may be an important contribution to changes in intracellular ion balance and the resultant pathophysiology of the disorder. The purpose of this study was to examine the effect of sepsis on the Na(+)-K(+)-ATPase in the isolated perfused rat heart using 133Cs+ nuclear magnetic resonance (NMR). Cs+ is a K+ analog, and 133Cs-NMR offers the opportunity to examine Na(+)-K(+)-ATPase activity in the intact organ via tracer kinetics. Sepsis was induced in halothane-anesthetized male Sprague-Dawley rats using the cecal ligation and perforation (CLP) model. Twenty-four to thirty-six hours after surgery, hearts from CLP or sham-operated rats were perfused with Krebs-Henseleit buffer containing 1.25 mM Cs+. The influx rate constant for Cs+ was decreased by 24% in septic rat hearts, i.e., 0.25 +/- 0.08 (SD) min 1 for controls and 0.19 +/- 0.04 (SD) min-1 for septic animals (P = 0.003). There was no difference for Cs+ efflux [0.005 +/- 0.001 (SD) min-1 for controls and 0.005 +/- 0.002 (SD) min-1 for septic animals; P = 0.8]. These results are consistent with an inhibition of the Na(+)-K(+)-ATPase pump during sepsis/endotoxemia. A decrease in the activity of the Na(+)-K(+)-ATPase pump may be responsible for or contribute to the changes in [Na+]i and [K+]i during the disorder.