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The Tradescantia micronucleus test is a sensitive bioassay for mutagenesis that may be employed both under field and laboratory conditions. This test has been standardized mostly on the basis of the results obtained with clone 4430. However, this clone is not well adapted to tropical weather, frequently showing problems with growth and flowering. In addition, it is attacked by parasites and insects, a fact that limits its use in field studies aiming at the biomonitoring of air pollution. In the city of São Paulo, Tradescantia pallida (Rose) Hunt. var. purpurea Boom is widely distributed as an ornamental plant in gardens and along roadsides and streets, mostly because of its natural resistance and its easy propagation. In this report, we present dose-response curves indicating that the sensitivity of T. pallida and clone 4430 to X-radiation (1, 10, 25 and 50 cGy) is similar. The results confirm our previous suggestion that T. pallida represents a good alternative for in situ mutagenesis testing in tropical regions, especially biomonitoring studies in which the exposure conditions may not be fully controllable.

This study was designed to determine the clastogenicity of particulate matter (aerodynamic diameter smaller than 10 microm) in the urban polluted air in the city of São Paulo. The Tradescantia-micronucleus (Trad-MCN) assay was used throughout this study to evaluate the clastogenicity of the extracts of the particulate matter. Tradescantia pallida (Rose) Hunt. cv. purpurea, an indigenous cultivar, was used in the Trad-MCN assay. The efficacy of this plant material for the Trad-MCN assay was validated with dose-response studies using formaldehyde and beta radiation. Dose-response curves were established with these known mutagens. The extracts of the PM10 particles at concentrations between 5 and 50 ppm induced a dose-related increase in MCN frequencies. The results indicate that T. pallida is equally sensitive to mutagens as the standard Tradescantia clone 4430 or 03 and the particulate matter in the urban air are clastogenic to the chromosomes of this plant. Inhalation of these particles by urban dwellers may affect their health by inducing similar genetic damage., (Copyright 1999 Elsevier Science B.V.)

This work was designed to develop a simple method based on the frog palate preparation to study the effects of hydrogen peroxide (H2O2) on ciliated epithelium. For this purpose, five sets (n = 10 per set) of frog palate preparations (Rana catesbeiana) were studied during 35 min after immersion in increasing concentrations of H2O2: 1, 8, 16, 32, and 64 microM. The effects of H2O2 on ciliated epithelium were assessed by measuring transepithelial potential difference (PD) and mucociliary transport (MT). Measurements were performed at 5-min intervals. In addition, the palates submitted to the 64 microM dose were immersed in Ringer's solution and followed by another 30 min to assess the possible recovery after maximal injury. Transepithelial potential difference (PD) was measured by means of agar-filled microelectrodes connected to the high input of a grounded electrometer. Mucociliary transport (MT) was determined by directly monitoring the movement of autologous mucus along the palate surface. Significant decrease in MT was observed in 16 microM and beyond and significant change in PD was observed in 32 microM and 64 microM. Palates submitted to 64 microM of H2O2 returned to their baseline levels of PD and MT within 30 min of recovery in Ringer's solution. In conclusion, the frog palate preparation was shown to be an efficient experimental tool to assess the deleterious effects of H2O2 on the ciliated epithelium.

The study was designed to investigate the influence of water pollution on gill apparatus. Specimens of Nile tilapia were collected from a polluted site in the São Paulo metropolitan area (Billings reservoir) and from a "clean" area. Fish from the polluted site showed a chronic inflammatory process in the distal region of the gill filaments, with epithelial hyperplasia. The raker length was increased and hypersecretion occurred, with a considerable volume of alcian blue-positive mucin on the epithelium of the rakers and a smaller volume of periodic acid-Schiff-positive mucin on the epithelial surface of the filaments. The rigidity of mucus in fish from the polluted site was increased, but no abnormalities in the viscosity to elasticity ratio or in mucus "wettability" were observed.

We describe some basic procedures for studying the properties of mucus. These techniques can be applied in both clinical and physiological studies to improve the understanding of the mechanisms related to epithelial defense in health and disease. Mucus collection--A major difficulty is the lack of simple and noninvasive methods for collecting normal mucus in sufficient quantity for later analysis. Physical properties of mucus--A. Rheology: Mucus exhibits both solid and liquid properties and the important factor governing the actual behavior is time. The magnetic microrheometer provides an elegant method for measuring rheological properties of microsamples. B. Adhesivity: It characterizes the forces of attraction between an adherent surface and an adhesive system and can be calculated by measuring the contact angle between a mucus drop and a surface. Mucus Transport--A. Mucus transport by cilia: Mucus is primarily cleared by the continuous ciliary beating, which can be studied using techniques such as the frog palate preparation as well as direct measurement, i.e., in situ mucus clearance. B. Cough clearance: It is essential for elimination of secretions in diseases leading to hypersecretory states. The cough machine simulates the flow-time profile of human coughing. Transepithelial potential difference--A potential difference exists between the epithelial surface and the submucosa and is the net result of the activity of the ion-transport system of the pulmonary epithelium. The potential can be measured using appropriate microelectrodes. Quantitative morphology--Methods may be used to characterize the epithelial surface condition that continuously changes during aggressive conditions.

The present work was designed to investigate whether the rheological determinants for nasal mucociliary transport are the same in the intact preparation (in situ), as they are when the mucus is added exogenously to the isolated, mucus-depleted frog palate (in vitro). We evaluated the association between estimators of mucociliary transport in both conditions and rheological parameters using multiple regression techniques. Two kinds of rats were used: (a) specific pathogen free (SPF) rats, representing the normal condition of respiratory epithelium; (b) non-SPF rats (NSPF), which have a chronic inflammatory process in the airways. In situ mucociliary clearance (MCC) was determined by measuring the displacement of charcoal particles placed in the nasal septum. In vitro mucociliary transport (MCT) of rat nasal mucus was measured using the isolated frog palate preparation. Mucus rheologic properties were determined by magnetic microrheometry, in oscillatory deformations performed at 1, 10 and 100 radians/sec. No differences were detected between SPF and NSPF rats in terms of rheological parameters. A decreased MCC was found in NSPF in comparison with the SPF group, but no differences were observed between groups in terms of MCT, as could be predicted by rheological data. When all animals were pooled, in situ transport was significantly associated with the viscosity/elasticity ratio, whereas in vitro transport was dependent on the total mechanical impedance of the mucus sample. In conclusion, in situ mucus transport is influenced by other rheological parameters than those associated with in vitro transportability.

Dynamic viscoelasticity measurements are required in many studies on biological fluids and they can be performed by determining the corresponding strain when a sinusoidal shear stress is applied to a sample. In several circumstances the amount of fluid that can be obtained for analysis in physiological conditions does not exceed microliters. In this context, the microrheometer technique is a useful approach to determine the dynamic rheological profile of the samples. However, the manual calculation of the desired parameters is tedious and time-consuming. This paper describes a menu-oriented program in order to facilitate its use by non-experts. The comparison between manual and computer-aided calculations demonstrated that the program reduced the time of measurement, and reduced intra- and interobserver variations. The program was developed on an IBM compatible PC in Microsoft C 5.1, and tested in a blind study to check the advantages in terms of time and reproducibility of the system verified by the concordance of two independent observers (interobserver influence) in two different occasions (intraobserver influence).

The relationship between mucus rheologic variables and in vitro ciliary transport was investigated in mucus samples collected from the upper airways of 30 Wistar rats. In vitro mucus transportability was determined by means of the frog palate preparation. Rheologic evaluation was done by measuring the rigidity modulus (log G*, representing the vectorial sum of viscosity and elasticity) and the loss tangent (tan delta, i.e. the ratio between viscosity and elasticity) at 1 and 100 radian/s using a magnetic microrheometer. The correlation between the rheologic variables and in vitro mucus transportability was made by stepwise multiple linear regression analysis, with frog palate transport rate considered as the dependent variable. A significant relationship was obtained between the rheologic parameters (log G* and tan delta) measured at 1 radian/s and the frog palate transport ratio. The relative speed of mucus samples was related to rheology according to the following relationship: rat/frog speed ratio = 1.666-0.434 log G*-0.331 tan delta, for G* and delta determined at 1 radian/s (multiple r = 0.666, p < 0.001). Transport rates predicted from the above formula gave a satisfactory fit to those observed in a second set of 30 rats. The present results indicate that the overall mucus impedance, as well as the ratio between viscosity and elasticity, are important in determining the efficiency of clearance. In addition, it was shown that measurements performed by applying relatively low frequency deformations are preferable for predicting ciliary transport.