Inhibition of development, swarming differentiation and virulence factors in Proteus mirabilis by an extract of Lithrea molleoides and its active principle (Z,Z)-5-(trideca-4',7'-dienyl)-resorcinol.

Antibacterial activity of Lithrea molleoides extract against Proteus mirabilis has been previously reported by our group. In the present study, the compound (Z,Z)-5-(trideca-4',7'-dienyl)-resorcinol (1) was isolated as its responsible active principle. The effects of the compound obtained and of L. molleoides extract on P. mirabilis growth and virulence factors were evaluated. Compound 1 showed MIC and MBC values of 4000 µg/ml. It was found that the extract, at four times the MIC, produced complete killing of the uropathogen at 2h from the beginning of the experiment, while the alkylresorcinol, at four times the MIC, produced the same effect after 24 h. Hemolysis was adversely affected in treatments with both products at 8 µg/ml, while hemagglutination was not altered. The whole extract induced complete autoaggregation of P. mirabilis at 2000 µg/ml, while compound 1 at the same concentration did not show this property. Swarming motility was delayed in treatments with the extract and with 1 at 1000 and 8 µg/ml, respectively, at 8h from the beginning of the assay. Complete inhibition of the phenomenon was still observed after 24 h when compound 1 was added at 125 µg/ml. These findings offer the possibility of new classes of antimicrobial medicines to tackle infections caused by P. mirabilis.

Solid-state characterization of two polymorphic forms of R-albuterol sulfate.

R-albuterol (levalbuterol) is a drug used for asthma therapy and some formulations of it are in solid dosage forms. The aim of this work was to describe and characterize two polymorphic modifications of R-albuterol sulfate by means of typical structure-sensitive analytical techniques such as X-ray powder diffraction, FT-IR spectroscopy, visual and microscopic inspection, and DSC. Substantial differences were observed between the solid-state properties of the crystals, confirming the existence of at least two polymorphic forms for R-albuterol sulfate: Form I and Form II.

Resolution of racemic albuterol via diastereomeric salts formation with di-p-toluoyl-D-tartaric acid.

A convenient method is described for the resolution of racemic Albuterol by selective crystallization of its di-p-toluoyl-D-tartrate salt. The separation resulted in a 38% yield of the (R)-enantiomer. Racemization of the (S)-enantiomer occurs in diluted H2SO4 at 100 degrees C in 80% yield. This racemic mixture was recycled to the diastereomer salt, in order to improve the overall yield. The (R)-Albuterol tartrate salt was decomposed in a sulfuric acid solution, and the (R)-Albuterol was isolated as its sulfate salt with 67% overall yield with 99.5% optical purity.

Fenoterol stimulation of sodium transport in the isolated toad skin: a beta adrenergic effect.

1. This work examines the effects of fenoterol on the isolated skin of the Chilean toad Pleurodema thaul. 2. A dose-dependent increase in the potential difference and in the short-circuit current of the skin was found. Furthermore, sodium potential (ENa) and sodium conductance (GNA) also rose. 3. The increase in bioelectric parameters was reversibly blocked by propranolol, was not significantly affected by reserpinization and was not calcium-dependent. 4. The skin response was significantly reduced in the presence of low Na+ in the outer bathing solution. Although the response was also reduced in the presence of isethionate Ringer's in the inner bathing solution, this reduction was less than the decrease in the presence of low Na+. 5. Fenoterol significantly increased toad skin oxygen consumption and net Na+ movement across the skin due to an increase in Na+ flux from mucosa to serosa. 6. These results show that fenoterol enhances active transport across the isolated toad skin probably through beta adrenergic stimulating effects which activate the sodium driving force, sodium conductance and net Na+ transepithelial flux.

Determination of the driving force for the sodium pump (ENa) and of active and passive conductances (GNa and Gsh) in isolated toad skin: influence of antidiuretic hormone.

1. Values of the sodium potential (ENa), active conductance (GNa) and passive conductance (Gsh) were measured in the isolated skin of the toad Pleurodema thaul placed in an Ussing chamber, and Isaacson's test was performed with 2,4,6-trieminopyrimidine (TAP) and with amiloride. 2. The numerical estimates obtained in the presence of TAP were ENa 122.85 +/- 15.17 mV, GNa 0.493 +/- 0.09 mS/cm2 and Gsh 1.145 +/- 0.23 mS/cm2. 3. After exposure to ADH these values were as follows: ENa 85.76 +/- 12.17 mV, GNa 1.191 +/- 0.20 mS/cm2 and Gsh 0.935 +/- 0.14 mS/cm2. 4. Addition of 0.5 x 10(2-) TAP produced a 53.90 +/- 5.10% decrease in transepithelial potential and a 37.90 +/- 4.90% fall in short-circuit current. 5. Exposure to ADH increased the transepithelial potential difference 34.20 +/- 13.20% and the short-circuit current to 78.00 +/- 20.50% above the control values. 6. Comparison of the efficiency and mechanism of action of TAP and amiloride in the determination of electrical parameters shows that both agents induce a similar decrease in Gsh, a finding which could indicate that TAP blocks toad skin apical membrane Na+ channels without affecting tight junction conductance.

Effects of antidiuretic hormone on the electrical properties of canine tracheal epithelium.

The canine tracheal epithelium causes net transport of Cl- from serosa to mucosa and of Na+ from mucosa to serosa. These processes are manifested by the short-circuit current which represents the algebraic sum of both fluxes. Antidiuretic hormone (ADH) alters the permeability of several epithelia. This work examines the effect and possible mechanism of action of ADH on the potential difference (PD), short-circuit current (ISC) and conductance (G) of canine tracheal epithelium. The epithelium was mounted in Ussing-type chambers containing Krebs-Henseleit solution at 37 degrees C (pH 7.4) and bubbled with 95% O2 and 5% CO2. PD was recorded on an Cole-Parmer recorder by means of the classical technique and ISC was recorded automatically using a voltage clamp. ADH (mucosal surface) significantly decreased PD, ISC and G in a dose-dependent fashion. Similar doses of ADH applied to the serosal side did not produce a significant effect. Angiotensin II (Agt II, serosal side) and amphotericin B (mucosal side) reversed the depressor effect of mucosal ADH. The effect of ADH was less when the Cl- serosa to mucosa movement was blocked by pretreatment with serosal furosemide. It is considered that ADH blocks apical membrane Cl-channels and that angiotensin reverses this effect by increasing apical membrane permeability to Cl-. Amphotericin B reverses the effect of ADH possibly by increasing Na+ absorption.