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BACKGROUND: The mechanism of transient global amnesia (TGA) is not clear. Attempting to support the hypothesis that retrograde venous hypertension causing cerebral venous ischemia plays a role in the pathogenesis of TGA, the authors used cranial three-dimensional time-of-flight (TOF) MR angiography (MRA) to detect a possible intracranial retrograde venous flow in TGA patients. METHODS: The frequency of abnormal venous signals on cranial three-dimensional TOF MRA was compared in 10 TGA patients with the signals in 50 age- and gender-matched normal individuals. In TGA patients with abnormal venous signals, other examinations (cerebral digital subtraction angiography, upper extremity digital subtraction venography [DSV], and thoracic inlet MRI) were performed to elucidate the etiology of these abnormal intracranial venous flow patterns. RESULTS: Abnormal venous signals on three-dimensional TOF MRA were found in five (50%) of the TGA patients and none of the control subjects (p < 0.001). Compression leading to occlusion of the left brachiocephalic vein by the sternum and aorta during regular breathing, as depicted by upper extremity DSV and thoracic inlet MRI, occurred consistently among these five TGA patients with abnormal venous signals. CONCLUSIONS: Retrograde intracranial venous flow caused by left brachiocephalic vein occlusion was found only in patients with transient global amnesia (TGA). This result suggests that TGA patients may have an underlying impairment of cerebral venous outflow that increases their vulnerability to TGA attack.

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The main objective of this study was to examine the efficacy and capacity of using constructed wetlands on industrial pollutant removal. Four parallel pilot-scale modified free water surface (FWS) constructed wetland systems [dimension for each system: 4-m (L)x1-m (W)x1-m (D)] were installed inside an industrial park for conducting the proposed treatability study. The averaged influent contains approximately 170 mg l(-1) chemical oxygen demand (COD), 80 mg l(-1) biochemical oxygen demand (BOD), 90 mg l(-1) suspend solid (SS), and 32 mg l(-1) NH(3)-N. In the plant-selection study, four different wetland plant species including floating plants [Pistia stratiotes L. (P. stratiotes) and Ipomoea aquatica (I. aquatica)] and emergent plants [Phragmites communis L. (P. communis) and Typha orientalis Presl. (T. orientalis)] were evaluated. Results show that only the emergent plant (P. communis) could survive and reproduce with a continuous feed of 0.4m(3)d(-1) of the raw wastewater. Thus, P. communis was used in the subsequent treatment study. Two different control parameters including hydraulic retention time (HRT) (3, 5, and 7d) and media [vesicles ceramic bioballs and small gravels, 1cm in diameter] were examined in the treatment study. Results indicate that the system with a 5-d HRT (feed rate of 0.4m(3)d(-1)) and vesicles ceramic bioballs as the media had the acceptable and optimal pollutant removal efficiency. If operated under conditions of the above parameters, the pilot-plant wetland system can achieve removal of 61% COD, 89% BOD, 81% SS, 35% TP, and 56% NH(3)-N. The treated wastewater meets the current industrial wastewater discharge standards in Taiwan.

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We performed the Queckenstedt's (Q)-test (compression over bilateral internal jugular veins) and a sham test on 33 patients with migraine attacks (coded as 1.1 based on headache classification proposed by International Headache Society (IHS)), 15 with migrainous attacks (IHS code 1.7), and 15 with tension-type headache (IHS code 2.1) in both supine and sitting positions. 'Migrainous headache' (code 1.7) was defined if the headache characteristics fulfilled all but one criteria for 'migraine without aura'. Migraine sufferers reported a marked increase in headache intensity after a 30-second Q-test in both supine and sitting positions. Aggravation was greater in the supine position. The intensity increase was not demonstrated in the sham test, or in patients with migrainous attacks or tension-type headaches after the Q-test. Patients with acute migraine thus appear more sensitive to increased cerebral venous pressure or intracranial pressure. The discrepancy of intensity changes between supine and sitting positions may reflect different amount of venous return through the internal jugular veins.

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Mushroom tyrosinase was immobilized on modified polystyrene- polyamino styrene (PSNH) and polymethylchloride styrene (PSCL)-to produce L-DOPA from L-tyrosine. Glutaraldehyde was used as an activating agent for the PSNH to immobilize the tyrosinase, and 10% (w/v) glutaraldehyde was optimal in conferring the highest specific activity (11.96 U/g) to the PSNH. Methylchloride on the PSCL was directly linked with the tyrosinase, and 1.5 mmol of Cl/g was optimal in attaining the specific activity of 17.0 U/g. The temperature and optimal acidity were, respectively, 60 degrees C and pH 5.5 for the PSNH, and 70 degrees C and pH 3.0 for the PSCL. In a 50-mL batch reactor working over 36 h, the L-DOPA production rate at 30 degrees C was 1.44 mg/(L x h) for the PSNH and 2.33 mg/(L x h) for the PSCL. The production rate over 36 h was 3.86 mg/(L x h) for the PSNH at 60 degrees C and 5.54 mg/(L x h) for the PSCL at 70 degrees C. Both of the immobilized enzymes showed a remarkable stability with almost no change in activity after being stored wet. The operational stability study indicated a 22.4% reduction in L-DOPA production for the PSNH and an 8.63% reduction for the PSCL over seven runs (each run was for 144 h at 30 degrees C) when the immobilized enzymes were used under turnover conditions. The immobilized tyrosinase was more stable on the PSCL than on the PSNH.

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BACKGROUND: Previous limited observations have suggested that atherosclerosis may affect the distensibility of the carotid sinus and then impair the baroreflex sensitivity (BRS). No studies have been done to compare the BRS and heart rate variability (HRV) in patients with carotid stenosis and normal controls. METHODS: A convenience-consecutive sample of 118 patients with transient ischemic attack or minor stroke 3 months to 1 year before (mean 6 months) who met the study criteria were referred to the neurovascular laboratory of the study hospital. Forty-three age-matched healthy adults were recruited as the normal controls. The inclusion criteria for participation were (1) no diabetes mellitus, (2) no history, symptoms or ECG signs of coronary artery disease or myocardial infarction, and (3) presence of carotid stenosis greater than or equal to 50%. The diagnosis of carotid stenosis was made using color-coded duplex ultrasound with published criteria. We categorized the patients into two groups: group 1 had moderate stenosis (50-75%) and group 2 had high-grade stenosis (75-99%). Instantaneous systolic blood pressure (SBP) and heart rate of all participants were assessed noninvasively using servo-controlled infrared finger plethysmography. The fluctuation in SBP as well as the interpulse interval (IPI) was divided into three components at specific frequency ranges by fast Fourier transform as high frequency (HF; 0.15-0.4 Hz), low frequency (LF; 0.04-0.15 Hz) and very low frequency (VLF; 0.004-0.04 Hz). The BRS was expressed as (1) transfer function with its magnitude in the HF and LF ranges, (2) BRS index alpha, and (3) regression coefficient by sequence analysis. The HRV was expressed as total power and power in the three frequency ranges (HF, LF and VLF). RESULTS: The final analysis included 99 patients (mean age 72 +/- 6 years, 79 male) and 43 healthy controls (mean age 68 +/- 7 years, 30 male). Forty-three patients were classified as group 1 (stenosis 50-75%) and 56 as group 2 (stenosis 75-99%). There was no significant difference in the IPI between patients and controls (p value = 0.8637). We observed a significant decrease in all three HRV components (VLF, LF and HF) in the patients; however, there were no differences between the two patient groups with various degrees of stenosis. All the indices of BRS, including the magnitude of SBP-IPI transfer function at LF and HF, the computed BRS index alpha and the regression coefficient of sequence analysis, revealed similar results. Patients exhibited a significant reduction in the BRS (p < 0.001) compared with controls, and no difference was found between the two groups of patients. CONCLUSIONS: Our study linked significant carotid stenosis to two important autonomic markers (BRS and HRV) that may have prognostic value for patients with cardiovascular events. Further prospective studies are needed to explore whether or not the decreased BRS and HRV can be predictors for poor cardiovascular prognosis, or even for shortened life span in general, in patients with significant carotid stenosis.

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The effectiveness of precipitation or coagulation technology to treat commercial egg processing plant wastewater, using such coagulants as lignosulfonate, bentonite, carboxymethylcellulose, and ferric chloride, was evaluated. For simulated and industrial waste-water, chemical oxygen demand, turbidity, and total solids were reduced over 90, 97, and 95%, respectively, for all coagulants tested. Protein and fat recoveries were over 95% for all coagulants. The optimal coagulant concentration for maximum by-product recovery depended on initial wastewater concentrations of protein, total solids, and fat. The dried by-products contained high concentrations of protein (30 to 50%) and fat (30 to 40%) and had similar essential amino acid profiles as standard proteins from the United Nations Food and Agricultural Organization (FAO). The relative protein digestibilities of each recovered solid (carboxymethycellulose, lignosulfonate, bentonite, and ferric chloride) and corn meal relative to a liquid whole egg standard were approximately 80, 90, 60, 30, and 56%, respectively. These compositional and in vitro digestibility studies suggest that the recovered by-products could be useful as livestock feed ingredients or for other applications.

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Sodium salts of medium-chain fatty acids, sodium caprate (C10) in particular, have been used as absorption-enhancing agents to promote transmucosal drug absorption. In this study, we conducted both in vitro and in vivo experiments to investigate the effects of C10 on intestinal permeabilities and mucosal morphology. Mucosal addition of C10 (13-25 mM) reduced the transepithelial electric resistance (TEER) of cultured monolayers of the human intestinal cell line Caco-2 by 40-65% and, upon removal of C10, a marked tendency of TEER recovery was recorded. C10 added mucosally at 13-50 mM increased the transports of mannitol and polyethylene glycol (PEG) 900 across Caco-2 in a dose-dependent manner. In contrast, the transport of a model D-decapeptide was maximally enhanced with 20-25 mM C10. No noticeable morphological alteration of the Caco-2 monolayers was observed after a 1-h mucosal pretreatment with C10. Co-delivery with C10 (0.05-0.5 mmol/kg) into the rat terminal ileum increased the D-decapeptide bioavailability (BA) dose-dependently. With 0.5 mmol/kg C10 co-administered, D-decapeptide percent BA was elevated from 2 to 11%. Following a 1-h incubation with 0.5 mmol/kg C10 (in liquid or powder form) non-invasively delivered into the rectal lumen, no signs of histological change in the rectal mucosa were detected. These results demonstrate that C10 can promote intestinal absorption of a small peptide without causing detrimental alterations of the intestinal mucosa. C10 thus seems to be a good candidate as an enhancing agent for improving the oral BA of small therapeutic peptides.

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DS-1, a modified Quillaja saponin, has recently been shown to promote the absorption of insulin and aminoglycoside antibiotics via the ocular and nasal route. The purpose of this study is to investigate the effect of DS-1 on intestinal permeability, the mechanism of its action, and reversibility of the effect. The permeation-enhancing activity of DS-1 was evaluated in cultured monolayers of the Caco-2 intestinal epithelial cells by examining its effect on the transepithelial electric resistance (TEER) and on transport of mannitol and a model D-decapeptide. Mucosal addition of DS-1 promptly reduced the TEER of the Caco-2 monolayers, and a propensity of recovery of the TEER was observed upon its removal. DS-1 added at 0.01-0.1% (w/v) increased the transports of both mannitol and D-decapeptide in a dose-dependent manner; a relatively "flat" concentration-dependence was seen at 0.1-0.2%. Visualization studies conducted by confocal laser scanning microscopy (CLSM) seem to suggest that DS-1 enhances the Caco-2 permeability mainly via a transcellular route. Histological examination failed to reveal noticeable morphological alterations in the cell monolayers pretreated with DS-1. The integrity of the Caco-2 monolayers, as assessed by their permeability to mannitol, was found to be recoverable following the mucosal pretreatment of DS-1. These results suggest that DS-1 is an efficacious intestinal permeation-enhancing agent with low adverse effect on the epithelial viability and barrier function.

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Long-chain acylcarnitines, such as palmitoylcarnitine chloride (PCC), are endogenous compounds which have been shown to increase intestinal transport of small hydrophilic compounds (including some pharmaceutical agents) through the paracellular pathway. However, the size range of the compounds whose absorption can be improved by PCC has not been fully investigated. In the present study, we systematically examined the effect of PCC on the transport rate of a series of hydrophilic fluorescent model compounds of varying molecular weights (0.3-71.2 kD) across cultured monolayers of the human intestinal epithelial cells Caco-2. Mucosal addition of 100 or 200 microM PCC resulted in comparable time-dependent decreases in the transepithelial electric resistance (T1/2, approximately 15 min). PCC addition induced a striking increase in the transport of sodium fluorescein (Flu-Na; 0.3 kD) and a slight or moderate increase in transports of fluorescent compounds of 0.6-11 kD. The effect of PCC on transport of compounds with molecular weights of > or = 17 kD appeared to be negligible. Examination by confocal laser scanning microscopy clearly revealed dilated paracellular spaces in Caco-2 monolayers which had been mucosally pretreated with PCC, confirming that PCC increases intestinal permeability by opening a paracellular transport pathway. Our results suggest that PCC is particularly effective in enhancing intestinal absorption of small hydrophilic compound like Flu-Na and may also have limited use in promoting the transport of compounds of < or = 10 kD.

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BACKGROUND: Leflunomide, a novel immunosuppressive drug, prolongs experimental graft survival effectively and has been well tolerated in patients with rheumatoid arthritis. A77 1726, the active metabolite of leflunomide, inhibits lymphocyte proliferation in vitro. This study was conducted in Jurkat T cells to investigate the effects of A77 1726 on signal transduction pathways initiated by ligands of the T-cell receptor CD3 complex and to evaluate the effects of A77 1726 on nucleotide biosynthesis. METHODS: Tritiated thymidine incorporation and cell counts quantitated cell proliferation. Spectrofluorescence of Indo/AM dye measured intracellular Ca2+ mobilization. A luciferase assay quantitated interleukin-2 gene promoter activity in stimulated cells transfected with an interleukin-2 promoter-luciferase gene construct. Pyrimidine and purine nucleosides were used to assess antagonism of the antiproliferative activity of A77 1726. RESULTS: (1) A77 1726 dose-dependently inhibited the proliferation of Jurkat T cells (inhibitory concentration of 50% = 6 mumol/L); (2) A77 1726 did not decrease mobilization of intracellular Ca2+ stimulated by phytohemagglutinin or anti-CD3 monoclonal antibody; (3) A77 1726 did not inhibit interleukin-2 gene promoter activity in cells stimulated with ionomycin plus phorbol myristate acetate; (4) inhibition of cell proliferation by A77 1726 was antagonized by addition of uridine, cytidine, or 2(+)-deoxycytidine; (5) addition of uridine 24 hours after treatment with A77 1726 antagonized inhibition of proliferation; (6) A77 1726 was not antagonized by 2'-deoxyuridine, thymidine, adenosine, or guanosine. CONCLUSIONS: (1) A77 1726 inhibited Jurkat T-cell proliferation without inhibiting T-cell receptor-mediated signal transduction events, including tyrosine kinase-dependent intracellular Ca2+ mobilization and activation of the interleukin-2 gene promoter; (2) the antiproliferative effects of A77 1726 on Jurkat T cells are primarily due to interruption of de novo pyrimidine nucleotide biosynthesis. These data provide evidence for a novel in vitro mechanism of the antiproliferative action of this immunosuppressant.

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Microsomal Ca(2+)-ATPase inhibitors such as thapsigargin (THG), cyclopiazonic acid (CPA) and 2,5-di-(tert-butyl)-1,4-hydroquinone (DBHQ) have been shown to inhibit Ca2+ reuptake by the intracellular stores and increase cytosolic free Ca2+ ([Ca2+]i). DBHQ is a commercially available non-toxic synthetic compound chemically unrelated to THG and CPA. In this study, we tested the feasibility of utilizing DBHQ to improve Cl- secretion via the Ca(2+)-dependent pathway, in the cystic fibrosis (CF)-derived pancreatic epithelial cell line CFPAC-1. DBHQ stimulated 125I efflux and mobilized intracellular free Ca2+ in a dose-dependent manner. The maximal effects were seen at concentrations of 25-50 microM. DBHQ (25 microM) caused a short-term rise in [Ca2+]i in the absence of ambient Ca2+, and a sustained elevation of [Ca2+]i in cell monolayers bathed in the efflux solution (1.2 mM Ca2+), which was largely attenuated by Ni2+ (5 mM). Bath-application of DBHQ induced an outwardly-rectifying whole-cell Cl- current, which was abolished by pipette addition of BAPTA (5 mM) or CaMK [273-302] (20 microM), an inhibitory peptide of multifunctional Ca2+/calmodulin-dependent protein kinase (CaMKII). Pretreatment of monolayers of CFPAC-1 cells with DBHQ for 4-5 min significantly increased the Ca(2+)-independent or autonomous activity of CaMKII assayed in the cell homogenates. Thus, DBHQ appears to enhance Cl- channel activity via a Ca(2+)-dependent mechanism involving CaMKII. Pretreatment of CFPAC-1 cells with up to 50 microM DBHQ for 6 h did not cause any detectable change in cell viability and did not significantly affect the cell proliferation rate. These results suggest that appropriate selective microsomal Ca(2+)-ATPase inhibitors may be therapeutically useful in improving Cl- secretion in CF epithelial cells.

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Nitric oxide, which is produced by cytokine-activated mononuclear cells, is thought to play an important role in inflammation and immunity. While the function of nitric oxide as a direct cytotoxic effector molecule is well established, its function as a transducer molecule in immune cells is not. By use of whole-cell patch clamp recordings, we show that nitric oxide activates cystic fibrosis transmembrane conductance regulator CI- currents in normal human cloned T cells by a cGMP-dependent mechanism. This pathway is defective in cystic fibrosis-derived human cloned T cells. These findings not only delineate a novel transduction mechanism for nitric oxide but also support the hypothesis that an intrinsic immune defect may exist in cystic fibrosis.

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Cystic fibrosis (CF), a lethal disease common to Caucasians, is characterized by a defect in the CF transmembrane conductance regulator and the resulting defective cAMP-regulated Cl- secretion by epithelial cells. Clinical manifestations include both pancreatic and pulmonary insufficiency. Traditional therapeutic modalities address these problems with pancreatic enzyme replacement, vitamins and nutritional supplementation, antibiotics, and respiratory therapy. However, newer therapies directed at the specific underlying defects have emerged. In this review, we discuss agents that increase Cl- secretion via preserved Cl- secretory pathways, such as uridine triphosphate, or that enhance Na+ resorption, such as amiloride, thereby correcting altered airway secretions. We also discuss agents, including deoxyribonuclease (DNase), that directly reduce sputum viscosity. CF is an early target for in vivo gene therapy, since it is a monogenic autosomal recessive disease in which restoration of normal cAMP-regulated Cl- conductance can be achieved by complementation with a normal gene. The early clinical gene therapy therapy work, with gene introduction by both viral and nonviral vectors, is discussed.

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It has been suggested that P-glycoprotein (P-gp), an ATP-dependent transporter responsible for classical multidrug resistance, is also a volume-regulated chloride channel. We reexamined this hypothesis by use of whole-cell patch clamp recordings of three matched pairs of cell lines, which were either drug-sensitive or drug-resistant due to P-gp overexpression. We demonstrate here that volume-regulated chloride-selective currents can be induced in cells with or without P-gp expression. Overexpression of either P-gp or cystic fibrosis transmembrane conductance regulator, the protein product of the CF gene and another member of the ATP-dependent transporters, is associated with a hypotonicity-induced, rapid onset, transient current prior to onset of the volume-sensitive chloride-selective current, an apparent nonspecific effect related to the overexpression of an integral membrane protein. These results suggest that there is no relationship between P-gp and the chloride channel activated by cell swelling.

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1. P1 purinoceptor agonists like adenosine have been shown to stimulate Cl- transport in secretory epithelia. In the present study, we investigated whether P1 agonist-induced Cl- secretion is preserved in cystic fibrosis airway epithelium and which signalling mechanism is involved. The effects of purinoceptor agonists on Cl- secretion were examined in a transformed cystic fibrosis airway phenotype epithelial cell line, CFPEo-. 2. Addition of adenosine (ADO; 0.1-1 mM) markedly increased 125I efflux rate. The rank order of potency of purinoceptor agonists in stimulating 125I efflux was ADO > AMP > ADP approximately equal to ATP. A similar order of potency was seen in transformed cystic fibrosis nasal polyp cells, CFNPEo- (ADO > ATP > AMP > ADP). These results are consistent with the activation of Cl- secretion via a P1 purinoceptor. 3. The P1 agonists tested (at 0.01 and 0.1 mM) revealed a rank order of potency of 5'-N-ethylcarboxamine adenosine (NECA) > 2-chloro-adenosine (2-Cl-ADO) > R-phenylisopropyl adenosine (R-PIA). 4. The known potent A2 adenosine receptor (A2AR) agonist, 5'-(N-cyclopropyl) carboxamidoadenosine (CPCA, 2 microM) but not the A1 adenosine receptor agonist, N6-phenyl adenosine (N6-phenyl ADO, 10 microM) markedly increased 125I efflux rate (baseline, 5.9 +/- 2.0% min-1, + CPCA, 10.9 +/- 0.6% min-1; P < 0.01). The stimulant effect of CPCA (10 microM) was abolished by addition of the A2AR antagonist 3,7-dimethyl-1-propargylxanthine (DMPX) (100 microM; reported K(i) = 11 +/- 3 microM). These results favour the involvement of A2AR. 5. ADO (0.1-mM) and CPCA (2 microM) both induced a marked increase in intracellular [Ca2+] ([Ca2+]i); the effect of the latter was again abolished by pretreatment of the cells with DMPX. By contrast, N6-phenyl ADO did not affect [Ca2+]i. 6. In patch-clamp experiments, ADO (1 mM) induced an outwardly-rectified whole-cell Cl- current (baseline, 2.5 +/- 0.8 pA pF-1, + ADO, 78.4 +/- 23.8 pA pF-1; P < 0.02), which was largely inhibited in cells internally perfused with a selective inhibitory peptide of the multifunctional Ca2+/calmodulin-dependent protein kinase, CaMK [273-302] (20 microM), as compared to a control peptide, CaMK [284-302]. Addition of BAPTA (10 mM), a Ca2+ chelator, to the perfusion pipette also abolished the ADO-elicited Cl- current. 7. In conclusion, our results suggest that A2AR participates in regulation of airway C1 secretion via aCa2+-dependent signalling pathway, which involves CaMK and appears to be at least partially conserved in cystic fibrosis airway epithelial cells.

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Heat-stable enterotoxins (STa) produced by pathogenic bacteria induce profound salt and water secretion in the gut, leading to diarrhea. Recently, guanylin, an endogenous peptide with properties similar to STa, was identified. While STa and guanylin bind to the same receptor guanylyl cyclase and raise cell cGMP, the signaling mechanism distal to cGMP remains controversial. Here we show that STa, guanylin and cGMP each activate intestinal Cl- secretion, and that this is abolished by inhibitors of cAMP-dependent protein kinase (PKA), suggesting that PKA is a major mediator of this effect. These agents induce Cl- secretion only in cells expressing the wild-type CFTR, indicating that this molecule is the final common effector of the signaling pathway. The involvement of CFTR suggests a possible cystic fibrosis heterozygote advantage against STa-induced diarrhea.

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The effect of non-steroidal anti-inflammatory drugs (NSAIDs) on the electrical properties of primary cultures of dog tracheal epithelium has been studied. The cells used were grown with an air interface in a serum-free medium on membranes coated with human placental collagen. When mounted in Ussing chambers at 37 degrees C, mean values for the baseline short circuit current (Isc) and the transepithelial resistance of 65 tissue specimens from 18 dogs were 24.0 +/- 3.2 microA/cm2 and 458 +/- 128 omega.cm2, respectively. These tissues had been pretreated with amiloride to abolish active Na+ absorption. Under these conditions, the Isc value serves as a measure of active Cl- secretion. The results of this study revealed that the Isc across a cultured monolayer of trachea was attenuated by the tested NSAIDs, indomethacin, fulfenamic acid, mefenamic acid, aspirin, and acetaminophen, with Ki's that ranged from 6.0 x 10(-5) to 2.51 x 10(-3) M. Salicylic acid had no effect on baseline Isc. The Isc sensitivity sequence to the Cl- channel inhibitors tested was: fulfenamic acid >> indomethacin > mefenamic acid >> aspirin > acetaminophen > salicylic acid. The NSAIDs also significantly inhibited both the transient, Ca(2+)-dependent and the sustained, cAMP-dependent increases in Isc elicited by isoproterenol. Thus, the tested NSAIDs appeared to have an effect on the electrical properties of the cells. A similar effect of NSAIDs on ion transport across the human airway epithelium may help to reduce airway fluid secretion.

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1. Apical membrane potential (Va), transepithelial potential (VT), fractional apical voltage ratio (FVa = delta Va/delta VT), tissue resistance (RT), and intracellular Cl- (aiCl) and K+ (aiK) activities were measured in isolated gallbladders maintained between oxygenated bicarbonate-free physiological media (23 degrees C, pH 7.2 or 8.2) in a divided chamber. The basolateral membrane potential (Vb) was calculated from the measured values of Va and VT. 2. Cl- removal from the serosal medium (which should accelerate coupled basolateral KCl exit) significantly depolarized Vb, decreased aiCl, decreased FVa, increased RT, and attenuated the depolarization of Vb (delta Vb) induced by high K+ added to the serosal side. These changes are consistent with a decrease in the K(+)-conductance of the basolateral membrane (gbK). 3. Addition of furosemide (an inhibitor of KCl cotransport) to the serosal medium induced significant increases in Vb, FVa, and high K(+)-induced delta Vb, indicating an increase in gbK. 4. In the presence of serosal furosemide, Cl- removal from the serosal medium did not significantly alter Vb, aiCl or delta Vb from their corresponding values when serosal Cl- was present. 5. Serosal furosemide had no significant effect on aiK and aiCl measured with double-barreled ion-selective microelectrodes. 6. These results suggest the possibility of a reciprocal relationship between gbK and the rate of basolateral KCl cotransport. This may contribute to the maintenance of aiK in gallbladder epithelial cells.

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Effects of niflumic acid (NFA) and flufenamic acid (FFA), the two nonsteroid anti-inflammatory agents recently reported to inhibit Cl- current in Xenopus oocytes, were examined in cultured monolayers of dog and cow trachea. Both agents showed potent inhibition to the short-circuit current (Isc), an index of magnitude of transepithelial Cl- secretion, with values of Ki of 0.02 (for NFA) and 0.06 (for FFA) mM, respectively. The sensitivity sequence of Isc to the Cl- channel inhibitors tested was NFA > FFA > diphenylamine-2-carboxylate (DPC) >> anthracene-9-carboxylate (A9C). Thus, NFA and FFA are so far the most potent commercially available Cl- channel inhibitors tested in Cl(-)-secreting epithelia. The sensitivity sequence of 36Cl uptake to the above Cl- channel inhibitors in Xenopus laevis oocytes was found to be identical to the cultures of trachea. This seems to imply that the membrane Cl- channels of Xenopus oocytes are functionally similar to that identified in mammalian Cl(-)-secreting epithelia.

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