ANO1 (TMEM16A) in pancreatic ductal adenocarcinoma (PDAC).

Pancreatic ductal adenocarcinoma (PDAC) has one of the worst survival rates of all cancers. ANO1 (TMEM16A) is a recently identified Ca(2+)-activated Cl(-) channel (CaCC) that is upregulated in several tumors. Although ANO1 was subject to extensive studies in the recent years, its pathophysiological function has only been poorly understood. The aim of the present study is to establish the significance of ANO1 in PDAC behavior and demarcate its roles in PDAC from those of the volume-regulated anion channel (VRAC). We performed qPCR and Western blot measurements on different PDAC cell lines (Panc-1, Mia PaCa 2, Capan-1, AsPC-1, BxPC-3) and compared the results to those obtained in a human pancreatic ductal epithelium (HPDE) cell line. All cancer cell lines showed an upregulation of ANO1 on mRNA and protein levels. Whole-cell patch-clamp recordings identified large Ca(2+) and voltage-dependent Cl(-) currents in PDAC cells. Using siRNA knockdown of ANO1 and three ANO1 inhibitors (T16Ainh-A01, CaCCinh-A01, and NS3728), we found that ANO1 is the main constituent of CaCC current in PDAC cells. We further characterized these three inhibitors and found that they had unspecific effects on the free intracellular calcium concentration. Functional studies on PDAC behavior showed that surprisingly inhibition of ANO1 did not influence cellular proliferation. On the other hand, we found ANO1 channel to be pivotal in PDAC cell migration as assessed in wound healing experiments.

Anoctamin 6 differs from VRAC and VSOAC but is involved in apoptosis and supports volume regulation in the presence of Ca2+.

Anoctamin 6 (ANO6), also known as TMEM16F, has been shown to be a calcium-activated anion channel with delayed calcium activation. The cellular function of ANO6 is under debate, and different groups have come to different conclusions about ANO6's physiological role. Although it is now quite well established that ANO6 is distinct from the volume-regulated anion channel, it is still unclear whether ANO6 or other anoctamins can be activated by cell swelling. In this study, we suggest that ANO1, ANO6, and ANO10 do not contribute to the volume-activated current in ANO-overexpressing HEK293 cells. Furthermore, knock-down of ANO6 in Ehrlich ascites tumor cells (EATC) and Ehrlich-Lettre ascites (ELA) did not decrease but instead significantly increased swelling-activated membrane currents. Knock-down of ANO6 in EATC did not reduce regulatory volume decrease (RVD) in the absence of extracellular calcium, whereas it significantly reduced RVD in the presence of calcium. Interestingly, we found that knock-down of ANO6 in ELA cells resulted in a decrease in cisplatin-induced caspase-3 activity, confirming earlier findings that ANO6 is involved in apoptosis. Finally, knock-down of ANO1 and ANO6 did not affect the volume-sensitive release of taurine in ELA cells. Thus, our data provide evidence that ANO6 cannot be activated directly by cell swelling unless Ca(2+) is present. We also conclude that ANO6 carries a current during RVD, provided extracellular calcium is present. Thus, swelling activation of ANO6 requires the presence of free calcium.

Membrane potential plays a dual role for chloride transport across toad skin.

The Cl- -current through toad skin epithelium depends on the potential in a way consistent with a potential-controlled Cl- permeability. Computer analysis of the Koefoed-Johnsen Ussing two-membrane model provided with constant membrane permeabilities indicates that the voltage- and time-dependent currents are not caused by a trivial Goldmand-type rectification and ion redistributions following transepithelial potential pertubations. Extended with a dynamic Cl- permeability in the apical membrane according to a Hodgkin-Huxley kinetic scheme, the model predicts voltage clamp data which closely resemble experimental observations. This extension of the classic frog skin model implies that the Cl- permeability is activated by a voltage change caused by the inward Na+ current through the apical membrane.

Heterogeneity of chloride channels in the apical membrane of isolated mitochondria-rich cells from toad skin.

The isolated epithelium of toad skin was disintegrated into single cells by treatment with collagenase and trypsine. Chloride channels of cell-attached and excised inside-out apical membrane-patches of mitochondria-rich cells were studied by the patch-clamp technique. The major population of Cl- channels constituted small 7-pS linear channels in symmetrical solutions (125 mM Cl-). In cell-attached and inside-out patches the single channel i/V-relationship could be described by electrodiffusion of Cl- with a Goldmann-Hodgkin-Katz permeability of, PCl = 1.2 x 10(-14) - 2.6 x 10(-14) cm3. s-1. The channel exhibited voltage-independent activity and could be activated by cAMP. This channel is a likely candidate for mediating the well known cAMP-induced transepithelial Cl- conductance of the amphibian skin epithelium. Another population of Cl- channels exhibited large, highly variable conductances (upper limit conductances, 150-550 pS) and could be activated by membrane depolarization. A group of intermediate-sized Cl(-)-channels included: (a) channels (mean conductance, 30 pS) with linear or slightly outwardly rectifying i/V-relationships and activity occurring in distinct "bursts," (b) channels (conductance-range, 10-27 pS) with marked depolarization-induced activity, and (c) channels with unresolvable kinetics. The variance of current fluctuations of such "noisy" patches exhibited a minimum close to the equilibrium-potential for Cl-. With channels occurring in only 38% of sealed patches and an even lower frequency of voltage-activated channels, the chloride conductance of the apical membrane of mitochondria-rich cells did not match quantitatively that previously estimated from macroscopic Ussing-chamber experiments. From a qualitative point of view, however, we have succeeded in demonstrating the existence of Cl-channels in the apical membrane with features comparable to macroscopic predictions, i.e., activation of channel gating by cAMP and, in a few patches, also by membrane depolarization.

Patch clamp on the luminal membrane of exocrine gland acini from frog skin (Rana esculenta) reveals the presence of cystic fibrosis transmembrane conductance regulator-like Cl- channels activated by cyclic AMP.

Chloride channels in the luminal membrane of exocrine gland acini from frog skin (Rana esculenta) constituted a single homogeneous population. In cell-attached patches, channels activated upon exposure to isoproterenol, forskolin, or dibutyryl-cAMP and isobutyl-1-methyl-xanthine rectified in the outward direction with a conductance of 10.0 +/- 0.4 pS for outgoing currents. Channels in stimulated cells reversed at 0 mV applied potential, whereas channels in unstimulated cells reversed at depolarized potentials (28.1 +/- 6.7 mV), indicating that Cl- was above electrochemical equilibrium in unstimulated, but not in stimulated, cells. In excised inside-out patches with 25 mM Cl- on the inside, activity of small (8-pS) linear Cl--selective channels was dependent upon bath ATP (1.5 mM) and increased upon exposure to cAMP-dependent protein kinase. The channels displayed a single substate, located just below 2/3 of the full channel amplitude. Halide selectivity was identified as PBr > PI > PCl from the Goldman equation; however, the conductance sequence when either halide was permeating the channel was GCl > GBr >> GI. In inside-out patches, the channels were blocked reversibly by 5-nitro-2-(3-phenylpropylamino)benzoic acid, glibenclamide, and diphenylamine-2-carboxylic acid, whereas 4, 4-diisothiocyanatostilbene-2,2-disulfonic acid blocked channel activity completely and irreversibly. Single-channel kinetics revealed one open state (mean lifetime = 158 +/- 72 ms) and two closed states (lifetimes: 12 +/- 4 and 224 +/- 31 ms, respectively). Power density spectra had a double-Lorentzian form with corner frequencies 0.85 +/- 0.11 and 27.9 +/- 2.9 Hz, respectively. These channels are considered homologous to the cystic fibrosis transmembrane conductance regulator Cl- channel, which has been localized to the submucosal skin glands in Xenopus by immunohistochemistry (Engelhardt, J.F., S.S. Smith, E. Allen, J.R. Yankaskas, D.C. Dawson, and J.M. Wilson. 1994. Am. J. Physiol. 267: C491-C500) and, when stimulated by cAMP-dependent phosphorylation, are suggested to function in chloride secretion.

Proton pump activity of mitochondria-rich cells. The interpretation of external proton-concentration gradients.

We have hypothesized that a major role of the apical H(+)-pump in mitochondria-rich (MR) cells of amphibian skin is to energize active uptake of Cl- via an apical Cl-/HCO3(-)-exchanger. The activity of the H+ pump was studied by monitoring mucosal [H+]-profiles with a pH-sensitive microelectrode. With gluconate as mucosal anion, pH adjacent to the cornified cell layer was 0.98 +/- 0.07 (mean +/- SEM) pH-units below that of the lightly buffered bulk solution (pH = 7.40). The average distance at which the pH-gradient is dissipated was 382 +/- 18 microns, corresponding to an estimated "unstirred layer" thickness of 329 +/- 29 microns. Mucosal acidification was dependent on serosal pCO2, and abolished after depression of cellular energy metabolism, confirming that mucosal acidification results from active transport of H+. The [H+] was practically similar adjacent to all cells and independent of whether the microelectrode tip was positioned near an MR-cell or a principal cell. To evaluate [H+]-profiles created by a multitude of MR-cells, a mathematical model is proposed which assumes that the H+ distribution is governed by steady diffusion from a number of point sources defining a set of particular solutions to Laplace's equation. Model calculations predicted that with a physiological density of MR cells, the [H+] profile would be governed by so many sources that their individual contributions could not be experimentally resolved. The flux equation was integrated to provide a general mathematical expression for an external standing [H+]-gradient in the unstirred layer. This case was treated as free diffusion of protons and proton-loaded buffer molecules carrying away the protons extruded by the pump into the unstirred layer; the expression derived was used for estimating stationary proton-fluxes. The external [H+]-gradient depended on the mucosal anion such as to indicate that base (HCO3-) is excreted in exchange not only for Cl-, but also for Br- and I-, indicating that the active fluxes of these anions can be attributed to mitochondria-rich cells.

A mathematical model of solute coupled water transport in toad intestine incorporating recirculation of the actively transported solute.

A mathematical model of an absorbing leaky epithelium is developed for analysis of solute coupled water transport. The non-charged driving solute diffuses into cells and is pumped from cells into the lateral intercellular space (lis). All membranes contain water channels with the solute passing those of tight junction and interspace basement membrane by convection-diffusion. With solute permeability of paracellular pathway large relative to paracellular water flow, the paracellular flux ratio of the solute (influx/outflux) is small (2-4) in agreement with experiments. The virtual solute concentration of fluid emerging from lis is then significantly larger than the concentration in lis. Thus, in absence of external driving forces the model generates isotonic transport provided a component of the solute flux emerging downstream lis is taken up by cells through the serosal membrane and pumped back into lis, i.e., the solute would have to be recirculated. With input variables from toad intestine (Nedergaard, S., E.H. Larsen, and H.H. Ussing, J. Membr. Biol. 168:241-251), computations predict that 60-80% of the pumped flux stems from serosal bath in agreement with the experimental estimate of the recirculation flux. Robust solutions are obtained with realistic concentrations and pressures of lis, and with the following features. Rate of fluid absorption is governed by the solute permeability of mucosal membrane. Maximum fluid flow is governed by density of pumps on lis-membranes. Energetic efficiency increases with hydraulic conductance of the pathway carrying water from mucosal solution into lis. Uphill water transport is accomplished, but with high hydraulic conductance of cell membranes strength of transport is obscured by water flow through cells. Anomalous solvent drag occurs when back flux of water through cells exceeds inward water flux between cells. Molecules moving along the paracellular pathway are driven by a translateral flow of water, i.e., the model generates pseudo-solvent drag. The associated flux-ratio equation is derived.

Endogenous chloride channels of insect sf9 cells. Evidence for coordinated activity of small elementary channel units.

The endogenous Cl- conductance of Spodoptera frugiperda (Sf9) cells was studied 20-35 h after plating out of either uninfected cells or cells infected by a baculovirus vector carrying the cloned beta-galactosidase gene (beta-Gal cells). With the cation Tris+ in the pipette and Na+ in the bath, the reversal potential of whole-cell currents was governed by the prevailing Cl- equilibrium potential and could be fitted by the Goldman-Hodgkin-Katz equation with similar permeabilities for uninfected and beta-Gal cells. In the frequency range 0.12 < f < 300 Hz, the power density spectrum of whole-cell Cl- currents could be fitted by three Lorentzians. Independent of membrane potential, >50% of the total variance of whole-cell current fluctuations was accounted for by the low frequency Lorentzian (fc = 0.40 +/- 0.03 Hz, n = 6). Single-Cl- channels showed complex gating kinetics with long lasting (seconds) openings interrupted by similar long closures. In the open state, channels exhibited fast burst-like closures. Since the patches normally contained more than a single channel, it was not possible to measure open and closed dwell-time distributions for comparing single-Cl- channel activity with the kinetic features of whole-cell currents. However, the power density spectrum of Cl- currents of cell-attached and excised outside-out patches contained both high and low frequency Lorentzian components, with the corner frequency of the slow component (fc = 0.40 +/- 0.02 Hz, n = 4) similar to that of whole-cell current fluctuations. Chloride channels exhibited multiple conductance states with similar Goldman-Hodgkin-Katz-type rectification. Single-channel permeabilities covered the range from approximately 0.6.10(-14) cm5/s to approximately 6.10(-14) cm3/s, corresponding to a limiting conductance (gamma 150/150) of approximately 3.5 pS and approximately 35 pS, respectively. All states reversed near the same membrane potential, and they exhibited similar halide ion selectivity, P1 > PCl approximately PBr. Accordingly, Cl- current amplitudes larger than current flow through the smallest channel unit resolved seem to result from simultaneous open/shut events of two or more channel units.

The role of antibacterial prophylaxis in urologic surgery.

The possible origins of post-TUR bacteriuria (urethral flora, prostate, bladder tumors, or contaminated irrigating fluids or instruments) are discussed. A review of the literature on antibacterial prophylaxis during TUR led to a positive assessment of its value. Preferred agents for prophylaxis of urinary tract infections include cephalosporins, extended-spectrum penicillins, ampicillin, and aminoglycosides. The value of antibacterial prophylaxis in open urologic surgery, such as prostatectomy, has not yet been clearly established, but studies have demonstrated significant benefit in patients undergoing transrectal needle biopsy of the prostate. New techniques of stone removal have not yet been thoroughly evaluated with respect to their potential for causing urinary tract infection and bacteremia. Since as many as 77 per cent of stones harbor bacteria, however, antibiotic prophylaxis is recommended. Nonurologic indications for antibacterial prophylaxis are outlined.

Infectious complications after instrumentation of urinary tract.

Urethral catheterization is the single most important predisposing factor in the development of nosocomial urinary tract infection. Infection rates, etiologies, and possible methods of prevention are reviewed. Cystoscopy may be followed by a transient bacteremia. It is recommended that patients with positive urine cultures who undergo diagnostic cystoscopy receive antibiotic prophylaxis, but this is not required in patients with sterile urine. The incidence of urinary tract infection following transurethral surgery in patients who have not been given prophylactic antibiotics ranges from 6 to 60 per cent. The value of antibacterial prophylaxis in TUR, is still somewhat controversial. The incidence of infection and the value of antibacterial prophylaxis in prostatic biopsy appear to be related to the technique (transperineal or transrectal) used for the biopsy. It is too early to assess the infection risks associated with relatively new urologic procedures, such as ureteroscopy and percutaneous nephrostomy. Nevertheless, any procedure that crushes or manipulates a potentially bacteria-harboring stone carries at least a theoretical risk of infection.

Amoxicillin/clavulanate in urinary tract infection.

A new beta-lactamase-stable oral antibiotic (Augmentin) has been found to be effective in the treatment of urinary tract infections (uncomplicated and complicated, recurrent, and nosocomial) and bacteriuria. The literature is reviewed.

Antimicrobial prophylaxis in urologic surgery.

Nosocomial urinary tract infections represent approximately 40 per cent of all nosocomial infections, thereby contributing considerably to secondary bacteremia and sepsis and possibly increased mortality. Urethral catheterization is the single most important predisposing factor in the development of urinary tract infection. Patients at increased risk of nosocomial infectious complications are the elderly, malnourished and debilitated, those with diabetes or prosthetic devices, and those on immunosuppressive therapy. About 75 per cent of nosocomial urinary tract infections are attributable to gram-negative bacteria, a disproportionate number of which when compared with community-acquired infections are caused by Proteus, Klebsiella, and Pseudomonas. There is enough evidence in the literature to support the use of prophylaxis in urologic surgery. Antimicrobial prophylaxis reduces the incidence of postoperative urinary tract infection. It does not seem to reduce the incidence of transient perioperative bacteremia but probably prevents the development of sepsis, thereby reducing the number of serious infective complications, the average hospital stay, and the associated total cost of treatment. To achieve adequate urine, blood, and tissue levels of the antimicrobial agent at the time of surgery, the drug should be given preoperatively. A short perioperative course represents sufficient prophylaxis. Regimens with combinations of beta-lactam and aminoglycoside antibiotics or single use of an extended spectrum beta-lactam antibiotic are acceptable for this purpose.

Iodine content in drinking water and other beverages in Denmark.

OBJECTIVE: To investigate the variation in iodine content in drinking water in Denmark and to determine the difference in iodine content between organic and non-organic milk. Further, to analyse the iodine content in other beverages. DESIGN AND SETTING: Tap water samples were collected from 41 evenly distributed localities in Denmark. Organic and non-organic milk was collected at the same time (twice summer and twice winter). Soft drinks, beers and juice were collected from different Danish producers and wine from different countries. All samples were analysed for iodine using inductively coupled mass spectrometry. RESULTS: Iodine in tap water varied from 2.1 to 30.2 microg/l; the iodine content was in general highest in the eastern part of Denmark and lowest in the western part of Denmark. Organic milk was found to have a lower iodine content than non-organic milk. CONCLUSIONS: Large geographical (and seasonal) variations in iodine concentrations were found in different beverages supplying an appreciable part of the iodine in the Danish diet. This knowledge is important when calculating the iodine intake from dietary intake studies. SPONSORSHIP: The 1991 Farmacy Foundation and Danish Veterinary and Food Administration. European Journal of Clinical Nutrition (2000) 54, 57-60

Studies of critical factors in the determination of arsenic in Standard Reference Materials of Marine origin by ETAAS:;NMKL(*) interlaboratory study.

A study to determine factors which are known to influence the electrothermal atomic absorption (ETAAS) determination of As has been performed. The study has been carried out using five sample solutions of marine Standard Reference Materials distributed to four participating laboratories. Uncoated and pyrolytically coated graphite tubes with L'vov platform and Ni and Pd/Mg as chemical modifiers have been tested. No differences in results have been obtained between AAS instruments equipped with Zeeman correction or deuterium arc background correction. Small differences in concentration levels of arsenic as well as in characteristic mass were found when chemical modifiers were compared. Pd/Mg will be recommended in order to avoid a contamination of the graphite furnace with nickel. The characteristic mass was improved by using pyrolytically coated graphite tubes with the L'vov platform compared with uncoated graphite tubes with the L'vov platform. In the interlaboratory study, the standard addition procedure will be recommended.

Morphology of the kidney in larvae of Bufo viridis (Amphibia, Anura, Bufonidae).

This study deals primarily with the morphology and ultrastructure of the pronephros in the green toad Bufo viridis during prometamorphosis when the pronephros and the developing mesonephros function simultaneously. Furthermore, the mesonephros was studied during pro- and postmetamorphosis with emphasis on the distal segments of the nephron. The paired kidneys consist of two cranial pronephroi immediately behind the gill region and two more caudal elongated mesonephroi. Each pronephros consists of a single convoluted tubule which opens into the coelom via three nephrostomes. This tubule is divided into three ciliated tubules, three proximal tubule branches, a common proximal tubule and a distal tubule, which in turn continues into the nephric duct. No intermediate segment is present. The length of the pronephric tubule is 12 mm, including the three branches of the ciliated tubules and proximal tubules. Primary urine is formed upon filtration from an external glomerulus, which is a convoluted capillary lined by podocytes, a specialization of the coelomic epithelium. From the coelom the filtrate is swept into the ciliated tubules. In the collecting duct system of the developing mesonephric nephron epithelial cells with conspicuous, apical osmiophilic granules appear in larvae of 9-10 mm. Heterocellularity of mixed intercalated (mitochondria rich) cells and principal cells is observed in the collecting duct system and nephric duct from a larval body length of 14 mm. As the proliferation of mitochondria-rich cells proceeds, the osmiophilic granules disappear and are completely absent from the adult amphibian mesonephros.

Sensitivities of selenite, selenate, selenomethionine and trimethylselenonium ion in aqueous solution and in blood plasma-ETAAS compared with ICP-MS.

Aqueous solutions and blood plasma spiked with selenite (Se(IV)), selenate (Se(VI)), selenomethionine (SeMet) or trimethylselenonium (TMSe) iodide were analyzed by Zeeman-corrected electrothermal atomic absorption spectrometry (ETAAS) using palladium as a chemical modifier, and by inductively coupled plasma mass spectrometry (ICP-MS). Using ETAAS, the sensitivities for Se(IV), SeMet and TMSe in aqueous solution were similar, whereas the sensitivity of Se(VI) was 63% of that value. In blood plasma, the ETAAS sensitivities of Se(IV) and Se(VI) were equal, whereas the sensitivities of SeMet and TMSe were 87 and 56%, respectively, of that value. In contrast, the ICP-MS sensitivities obtained for Se(VI), TMSe and SeMet were between 96 and 98% of that obtained for Se(IV) in aqueous solution and in blood plasma. It is concluded, that ICP-MS is superior to ETAAS as the problem of differences in sensitivity of the selenium species when using ETAAS are not prevalent when using the ICP-MS technique.

Absorption and retention of selenium from shrimps in man.

This study was undertaken to evaluate the bioavailability of selenium in shrimps, a possible good source of selenium, by measurements of the absorption and retention of selenium and the effects on plasma selenium concentration and glutathione peroxidase activity. Twelve healthy young subjects (9F and 3M) received 100 g of shrimps each day for six weeks in addition to their habitual diet. In the third week of the study a balance period was inserted in which the subjects received all food from the department and collected faeces and urine over 5 days. Blood samples were collected at commencement of the study, after 2, 4, and 6 weeks. The selenium intake increased from 39.4 +/- 15.3 microg/d to 127 +/- 5.5 microg/d with the addition of shrimps. The apparent absorption of selenium from shrimps was 83 +/- 4%. Faecal and urinary selenium excretion was 32.5 +/- 17.0 microg/d and 21.2 +/- 9.0 microg/d, respectively and the total retention of selenium was 3.1 +/- 1.1 mg. Plasma selenium concentrations were 95.2 +/- 9.7 microg/L and 101.5 +/- 9.7 microg/L before and after six weeks of shrimp intake, respectively (p<0.05). Plasma and erythrocyte glutathione peroxidase activities were not influenced by shrimp intake. Thus, despite the high absorption and retention, plasma selenium concentrations were only moderately affected by an increase in selenium intake of about 100 microg/d in the chemical forms found in shrimp. Whether the accumulation of selenium from shrimps in tissues may represent a potential hazard is to be further investigated.

Atmospheric deposition of trace elements around point sources and human health risk assessment. II. Uptake of arsenic and chromium by vegetables grown near a wood preservation factory.

Kale, lettuce, carrots and potatoes were grown in 20 experimental plots surrounding a wood preservation factory, to investigate the amount and pathways for plant uptake of arsenic and chromium. Arsenate used in the wood preservation process is converted to the more toxic arsenite by incineration of waste wood and is emitted into the atmosphere. Elevated concentrations of inorganic arsenic and chromium were found both in the test plants and in the soil around the factory. Multivariate statistical analysis of the results indicated that the dominating pathway of arsenic and chromium from the factory to the leafy vegetables grown nearby was by direct atmospheric deposition, while arsenic in the root crops originated from both the soil and the atmosphere. Consumption of vegetables grown near the source would result in an increased intake of inorganic arsenic, but the intake via the total diet was estimated to be below the provisional tolerable daily intake for inorganic arsenic established by FAO/WHO.

Atmospheric deposition of trace elements around point sources and human health risk assessment. I: Impact zones near a source of lead emissions.

The deposition of lead was monitored over 8 years in the area around a car battery factory north of Copenhagen, Denmark. The area also has heavy traffic. Deposition was measured by in-situ grown vegetables, transplant grass culture biomonitors, bulk deposition and soil samples. Three impact zones were identified by a multivariate statistical analysis. Within each zone, the total dietary intake of lead was estimated for adults and children as a percentage of the provisional tolerably weekly intake (PTWI), and as a result recommendation on restrictions in use of locally grown fruit and vegetables were given to the public. The pattern of lead deposition in the area during the period 1981-1988 was monitored and the amount of lead ingested via vegetables was toxically evaluated. Lead emission reduction measures introduced in the factory and in the traffic during the period produced significant reductions in lead deposition.

Quinolone penetration into canine vaginal and urethral secretions.

Four newer quinolones (amifloxacin, ciprofloxacin, enoxacin, norfloxacin) were administered to female dogs by intravenous infusion. Drug concentrations in plasma, urine, and vaginal and urethral secretion were determined by bioassay. All four quinolones penetrated into vaginal and urethral secretion in concentrations several times higher than the MIC against common urinary pathogens, ciprofloxacin and norfloxacin reaching concentrations exceeding the simultaneous plasma concentrations. Because of their favorable antibacterial spectra, new quinolones should be investigated clinically for the treatment of recurrent urinary tract infection and bacterial vaginitis.