Antibodies against ClC7 inhibit extracellular acidification-induced Cl⁻ currents and bone resorption activity in mouse osteoclasts.

The Cl⁻ channel/transporter ClC7 is crucial for osteoclastic bone resorption and might become a therapeutic target for osteoporosis. In this study, we raised anti-ClC7 polyclonal antibodies against three different peptide sequences, including G215, P249, and R286, which are the mutation regions found in autosomal dominant osteopetrosis type II patients and examined the effects of these antibodies on the ClC7 Cl⁻ current induced by extracellular acidification (acid-activated Cl⁻ current) using the whole-cell patch clamp technique and bone resorption activity in mouse osteoclasts. Intracellular dialysis of osteoclasts with antibodies to intracellular G215 (Ab-G215) and extracellular application of antibodies to extracellular P249 (Ab-P249) or R286 (Ab-R286) inhibited the acid-activated Cl⁻ current. These antibodies also suppressed the acid-activated Cl⁻ current in ClC7 overexpressing Raw264.7 cells; however, Cl⁻ currents evoked by hypotonic stimulation and the inherent inwardly rectifying K+ currents in mouse osteoclasts were unaffected by these antibodies. Furthermore, extracellularly applied Ab-P249 and Ab-R286 also reduced bone resorption activity. Our results demonstrate that these antibodies specifically block ClC7 in mouse osteoclasts. Thus, anti-ClC7 antibodies have potential promise for treatment of osteoporosis.

Mechanism and rate of middle ear fluid absorption.

Several mechanisms have been suggested to explain the clearance of fluids from the middle ear. These include a pumping action through the eustachian tube, mucociliary beating through the tube, outflow of water to the blood due to osmotic gradients and an active Na(+) transport driving water absorption. In order to assess these mechanisms, the middle ear cavity of paralyzed, ventilated (eustachian tube occluded) guinea pigs was filled with fluids varying in osmotic pressure (hypotonic, isotonic, hypertonic) to which a vertical tube was attached. The change in height of fluid in the tube was taken as a measure of changes in middle ear fluid volume. A greater fluid volume reduction was seen with the hypotonic (1/5 saline) solution. A small volume increase was observed with the hypertonic solution. These results provide evidence that in these experimental conditions, water absorption due to osmotic gradients can contribute to middle ear fluid clearance.

Carrier solutions for intraperitoneal chemotherapy.

Two pharmacokinetic problems appear to limit the effectiveness of intraperitoneal therapy: poor tumor penetration and nonuniform intraperitoneal distribution by the drug-containing solution. A better understanding of the pharmacodynamics of carrier solutions for intraperitoneal chemotherapy may impact on both these problems. Not only is the choice of drug important, but also the matching of the chosen drug to an appropriate carrier solution. It would appear that high molecular weight solutions offer a number of advantages over low molecular weight glucose and saline solutions for chemotherapeutic agents that have a high molecular weight with a slow peritoneal clearance. The use of carrier solutions of varying tonicity requires further investigation.

Transurethral resection syndrome: effect of the introduction into clinical practice of a new method for monitoring fluid absorption.

STUDY OBJECTIVE: To determine the effects of introduction of a new monitoring system for fluid absorption during transurethral resection of the prostate (TURP) using an irrigating solution containing 0.5% alcohol. STUDY DESIGN: Prospective clinical investigation, with implementation of statistical process control. SETTING: Inpatients for TURP at a major non-university teaching hospital. PATIENTS: 312 male ASA physical status I, II, III, and IV patients scheduled for TURP. INTERVENTIONS: Intraoperative breath alcohol levels were measured for detection of fluid absorption. MEASUREMENTS AND MAIN RESULTS: Calculation of the amount of fluid absorbed using measured breath alcohol values. Process variability (i.e., numbers of patients with significant fluid absorption) was defined by statistical process control tools. No trend change of prevalence of fluid absorption was noted until 150 procedures had been completed. Reduction of prevalence of significant fluid absorption was noted and no patients were treated postoperatively in the intensive care unit. No relevant side effects were seen in patients with significant fluid absorption. No mortality and no severe clinical morbidity was seen after the introduction of the new monitoring. CONCLUSION: Using an irrigating fluid marked with 0.5% ethanol resulted in a decreased prevalence of fluid absorption over time.

Validation of polyethylene glycol 3350 as a poorly absorbable marker for intestinal perfusion studies.

Polyethylene glycol (PEG) has been used as a poorly absorbable marker in intestinal perfusion studies, but there is controversy about the absorbability of PEG, particularly when glucose-sodium cotransport is occurring. Total intestinal perfusion studies were done in five normal humans using three solutions containing 1 g/liter PEG 3350 and designed to produce low rates of water absorption, high rates of water absorption, or high rates of glucose-sodium cotransport. Water absorption rates were calculated by traditional nonabsorbable marker equations and by a novel balance technique in which absorption was taken as the difference between the volumes of solution infused and recovered during steady-state conditions. Effluent PEG recovery was 99 +/- 4%, 109 +/- 2%, and 104 +/- 6% of the amount infused with each solution. Water absorption rates measured by use of PEG concentrations were similar to those calculated by the balance technique (r = 0.99). The complete recovery of PEG confirms the poor absorbability of PEG 3350, and the excellent agreement between techniques validates PEG as a poorly absorbed marker, even when glucose-sodium cotransport is occurring.

Access to rat kidney with minimal anesthesia and surgery: a new experimental model.

Chronic surgical explantation of the left rat kidney out of the abdominal cavity under the flank skin enabled easy access to the organ and ipsilateral urine collection under light chloralose anesthesia without virtually any surgical intervention. The glomerular filtration rate, urine flow and osmolality, sodium excretion, and medullary tissue hypertonicity were similar in the explanted and in the contralateral kidney, whereas p-aminohippurate clearance was 14% lower. The function of the explanted kidney was also compared with that of the kidney acutely exposed in rats under thiobutabarbital anesthesia and rendered euvolemic by isoncotic albumin infusion. Again, in both preparations renal function was comparable except that over time urine osmolality remained stable in the former and fell from 1,385 +/- 195 to 835 +/- 167 mosmol/kgH2O (P < 0.02) in the latter, indicating deterioration of urine concentration. Laser-Doppler probes could be easily applied in the explanted kidney to measure cortical and medullary blood flow. The new experimental model offers some advantages, both over studies using conscious rats and over experiments involving deep anesthesia and acute surgery.

Osmolar changes and neural activity in frog vestibular organs.

The effects of hypotonic and hypertonic solutions (the normal value was 240 mOsm) on posterior canal resting and evoked discharge were studied in isolated labyrinth preparations. Hypotonic solutions (60-180 mOsm) were obtained by reducing the perilymphatic NaCl content. Hypertonic solutions (300-420 mOsm) were obtained by adding to normal perilymphatic solutions suitable amounts of NaCl, glucose, sucrose, glycerol, mannitol and urea. The results demonstrated that any kind of receptor activity was inhibited by hypotonic solutions. On the contrary, hypertonic solutions produced different effects on resting and evoked activity. The resting discharge was, with the exception of urea, constantly increased whereas the evoked responses were constantly decreased by all the hypertonic solutions tested. The possible effects of media with changed osmolarity in Meniere's patients is also discussed.

Regional deposition of saline aerosols of different tonicities in normal and asthmatic subjects.

Nonisotonic aerosols are frequently used in the diagnosis and therapy of lung disease. The purpose of this work was to study the difference in the pattern of deposition of aerosols containing aqueous solutions of different tonicities. 99mTechnetium-diethyltriaminepentaacetic acid (99mTc-DTPA)-labelled saline aerosols, with mass median aerodynamic diameter 3.7-3.8 microns and geometric standard deviation 1.4, were inhaled under reproducible breathing conditions on two occasions. Hypotonic and hypertonic solutions were used in 11 normals subjects, isotonic and hypertonic solutions in 9 asthmatics. The regional deposition was quantified by a penetration index measured with the help of a tomographic technique. There was a small but significant increase (6.7%) in the penetration index of the hypotonic as compared to the hypertonic aerosols in the normal subjects. The region that was markedly affected was the trachea. The differences in the penetration of the isotonic and hypertonic aerosols in the asthmatics appeared to be strongly dependent on the state of the airways at the time of the study. These findings can be interpreted in terms of effects of growth or shrinkage of nonisotonic aerosols, as well as of airway narrowing, on regional deposition of aerosols. Tonicity of aerosols appears to affect their deposition both through physical and physiological mechanisms. This should be taken into account when interpreting the effects of inhaled aqueous solutions of various tonicities in patients in vivo.

Regional deposition of nebulized hypodense nonisotonic solutions in the human respiratory tract.

Deposition of nonisotonic therapeutic and diagnosis aerosols can cause changes in airway fluid composition and bronchoconstriction in sensitive subjects. "Hypodense" aerosols containing a relatively low concentration of droplets in the carrier air were used in the studies of regional deposition of radiolabelled nebulized solutions of hypo- and hypertonic saline, in order to investigate whether the number of droplets per volume of carrier can affect deposition. Solutions with and without 0.5% nedocromil sodium were nebulized in order to examine the effects of a potential modifier of the rates of heat and mass transfer. The deposition was quantified using penetration index (PI) calculated from images obtained by single photon emission computerized tomography (SPECT) in 11 healthy volunteers per study. There was an increase in the penetration index (10.9%, for the saline only; 15.5%, for the nedocromil study) of the hypotonic compared to the hypertonic aerosol, although the initial size distribution of both types of aerosols was very similar (mass median aerodynamic diameter (MMAD) 3.7 and 3.8 microns; geometric standard deviation (GSD) 1.8 and 1.5 for the hypo- and hypertonic aerosols, respectively). The present results confirm the effects of tonicity on deposition of aerosols found in a parallel study reported in this issue of the Journal. They also give support to the theory that, in addition to the concentration of the nebulized solutions, the number of droplets per volume of the carrier air is a factor affecting deposition of aqueous aerosols. The presence of 0.5% nedocromil sodium in the solutions did not appear to interfere with the processes of heat and water transfer in the airways.

The effect of solution non-ideality on membrane transport in three-dimensional models of the renal concentrating mechanism.

Previous models of the renal concentrating mechanism employ ideal approximations of solution thermodynamics for membrane transport calculation. In three-dimensional models of the renal medulla, predicted urine concentrations reach levels where these idealized approximations begin to break down. In this paper we derive equations that govern membrane transport for non-dilute solutions and use these equations in a three-dimensional model of the concentrating mechanism. New numerical methods were employed that are more stable than those employed previously. Compared to ideal solution models, the urea non-ideality tends to increase predicted osmolarities, whereas NaCl non-ideality decreases predictions.