A Comparison of the Effect of Lead (Pb) on the Slow Vacuolar (SV) and Fast Vacuolar (FV) Channels in Red Beet (Beta vulgaris L.) Taproot Vacuoles.

Little is known about the effect of lead on the activity of the vacuolar K+ channels. Here, the patch-clamp technique was used to compare the impact of lead (PbCl2) on the slow-activating (SV) and fast-activating (FV) vacuolar channels. It was revealed that, under symmetrical 100-mM K+, the macroscopic currents of the SV channels exhibited a typical slow activation and a strong outward rectification of the steady-state currents, while the macroscopic currents of the FV channels displayed instantaneous currents, which, at the positive potentials, were about three-fold greater compared to the one at the negative potentials. When PbCl2 was added to the bath solution at a final concentration of 100 µM, it decreased the macroscopic outward currents of both channels but did not change the inward currents. The single-channel recordings demonstrated that cytosolic lead causes this macroscopic effect by a decrease of the single-channel conductance and decreases the channel open probability. We propose that cytosolic lead reduces the current flowing through the SV and FV channels, which causes a decrease of the K+ fluxes from the cytosol to the vacuole. This finding may, at least in part, explain the mechanism by which cytosolic Pb2+ reduces the growth of plant cells.

Response of Two Crop Plants, Zea mays L. and Solanum lycopersicum L., to Diclofenac and Naproxen.

Among numerous contaminants, the ubiquitous occurrence of nonsteroidal anti-inflammatory drugs (NSAIDs) in the environment and their plausible harmful impact on nontarget organisms have made them one of the most important areas of concern in recent years. Crop plants can also potentially be exposed to NSAIDs, since the concentration of these pharmaceuticals is constantly rising in the surface water and soil. Our goal was to evaluate the stress response of two crop plants, maize and tomato, to treatment with selected NSAIDs, naproxen and diclofenac. The focus of the research was on the growth response, photosynthetic efficiency, selected oxidative stress factors (such as the H2O2 level and the rate of lipid peroxidation) as well as the total phenolic content, which represents the non-enzymatic protectants against oxidative stress. The results indicate that susceptibility to the NSAIDs that were tested is dependent on the plant species. A higher sensitivity of tomato manifested in growth inhibition, a decrease in the content of the photosynthetic pigments and a reduction in the maximum quantum efficiency of PSII and the activity of PSII, which was estimated using the Fv/Fm and Fv/F0 ratios. Based on the growth results, it was also possible to reveal that diclofenac had a more toxic effect on tomato. In contrast to tomato, in maize, neither the content of the photosynthetic pigments nor growth appeared to be affected by DFC and NPX. However, both drugs significantly decreased in maize Fv and Fm, which are particularly sensitive to stress. A higher H2O2 concentration accompanied, in most cases, increasing lipid peroxidation, indicating that oxidative stress occurred in response to the selected NSAIDs in the plant species that were studied. The higher phenolic content of the plants after NSAIDs treatment may, in turn, indicate the activation of defense mechanisms in response to the oxidative stress that is triggered by these drugs.

Effect of Auxin (IAA) on the Fast Vacuolar (FV) Channels in Red Beet (Beta vulgaris L.) Taproot Vacuoles.

In contrast to the well-studied effect of auxin on the plasma membrane K+ channel activity, little is known about the role of this hormone in regulating the vacuolar K+ channels. Here, the patch-clamp technique was used to investigate the effect of auxin (IAA) on the fast-activating vacuolar (FV) channels. It was found that the macroscopic currents displayed instantaneous currents, which at the positive potentials were about three-fold greater compared to the one at the negative potentials. When auxin was added to the bath solution at a final concentration of 1 µM, it increased the outward currents by about 60%, but did not change the inward currents. The imposition of a ten-fold vacuole-to-cytosol KCl gradient stimulated the efflux of K+ from the vacuole into the cytosol and reduced the K+ current in the opposite direction. The addition of IAA to the bath solution with the 10/100 KCl gradient decreased the outward current and increased the inward current. Luminal auxin reduced both the outward and inward current by approximately 25% compared to the control. The single channel recordings demonstrated that cytosolic auxin changed the open probability of the FV channels at the positive voltages to a moderate extent, while it significantly increased the amplitudes of the single channel outward currents and the number of open channels. At the positive voltages, auxin did not change the unitary conductance of the single channels. We suggest that auxin regulates the activity of the fast-activating vacuolar (FV) channels, thereby causing changes of the K+ fluxes across the vacuolar membrane. This mechanism might serve to tightly adjust the volume of the vacuole during plant cell expansion.

Role of auxin (IAA) in the regulation of slow vacuolar (SV) channels and the volume of red beet taproot vacuoles.

BACKGROUND: Auxin (IAA) is a central player in plant cell growth. In contrast to the well-established function of the plasma membrane in plant cell expansion, little is known about the role of the vacuolar membrane (tonoplast) in this process. RESULTS: It was found that under symmetrical 100 mM K+ and 100 µM cytoplasmic Ca2+ the macroscopic currents showed a typical slow activation and a strong outward rectification of the steady-state currents. The addition of IAA at a final concentration of 1 µM to the bath medium stimulated the SV currents, whereas at 0.1 and 10 µM slight inhibition of SV currents was observed. The time constant, τ, decreased in the presence of this hormone. When single channels were analyzed, an increase in their activity was recorded with IAA compared to the control. The single-channel recordings that were obtained in the presence of IAA showed that auxin increased the amplitude of the single-channel currents. Interestingly, the addition of IAA to the bath medium with the same composition as the one that was used in the patch-clamp experiments showed that auxin decreased the volume of the vacuoles. CONCLUSIONS: It is suggested that the SV channels and the volume of red beet taproot vacuoles are modulated by auxin (IAA).

Single and combined effects of Cd and Pb on the growth, medium pH, membrane potential and metal contents in maize (Zea mays L.) coleoptile segments.

The mechanisms of the toxic effects of Cd and Pb on plant cell growth are still poorly understood. In particular, little is known about their interactive effects, which usually occur in the environment. Moreover, the data that do exist in the literature are controversial. This study describes experiments that were performed with maize (Zea mays) coleoptile segments, which is a classical model system for studies of plant cell elongation growth. Cadmium and lead, which were added at 0.1 mM, reduced the endogenous and IAA-induced elongation growth of maize coleoptile cells. When both metals were added together or in sequence, their effect on IAA-induced growth was more toxic. The medium pH changes, which were measured simultaneously with growth, indicated that while Pb stopped IAA-induced proton extrusion, Cd only partially diminished it. Although Cd was generally more accumulated than Pb in the maize coleoptile segments, when IAA was added together with Pb, it significantly increased the accumulation of the metal. The short-term electrophysiological experiments showed that the addition of Cd caused the depolarisation of the membrane potential (Em), whereas Pb caused membrane hyperpolarisation. In the long-term electrophysiological experiments, it was found that the Cd-induced Em changes are complex. In conclusion, these results suggest that the effects of Cd and Pb as well as their combination on the elongation growth of maize coleoptile cells and the accumulation of the metals result, among others, from different ionic mechanisms by which each metal change the membrane potential of the cells.

Role of chloride ions in the promotion of auxin-induced growth of maize coleoptile segments.

BACKGROUND AND AIMS: The mechanism of auxin action on ion transport in growing cells has not been determined in detail. In particular, little is known about the role of chloride in the auxin-induced growth of coleoptile cells. Moreover, the data that do exist in the literature are controversial. This study describes experiments that were carried out with maize (Zea mays) coleoptile segments, this being a classical model system for studies of plant cell elongation growth. METHODS: Growth kinetics or growth and pH changes were recorded in maize coleoptiles using two independent measuring systems. The growth rate of the segments was measured simultaneously with medium pH changes. Membrane potential changes in parenchymal cells of the segments were also determined for chosen variants. The question of whether anion transport is involved in auxin-induced growth of maize coleoptile segments was primarily studied using anion channel blockers [anthracene-9-carboxylic acid (A-9-C) and 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS)]. In addition, experiments in which KCl was replaced by KNO3 were also performed. KEY RESULTS: Both anion channel blockers, added at 0·1 mm, diminished indole-3-acetic acid (IAA)-induced elongation growth by ~30 %. Medium pH changes measured simultaneously with growth indicated that while DIDS stopped IAA-induced proton extrusion, A-9-C diminished it by only 50 %. Addition of A-9-C to medium containing 1 mm KCl did not affect the characteristic kinetics of IAA-induced membrane potential changes, while in the presence of 10 mm KCl the channel blocker stopped IAA-induced membrane hyperpolarization. Replacement of KCl with KNO3 significantly decreased IAA-induced growth and inhibited proton extrusion. In contrast to the KCl concentration, the concentration of KNO3 did not affect the growth-stimulatory effect of IAA. For comparison, the effects of the cation channel blocker tetraethylammonium chloride (TEA-Cl) on IAA-induced growth and proton extrusion were also determined. TEA-Cl, added 1 h before IAA, caused reduction of growth by 49·9 % and inhibition of proton extrusion. CONCLUSIONS: These results suggest that Cl(-) plays a role in the IAA-induced growth of maize coleoptile segments. A possible mechanism for Cl(-) uptake during IAA-induced growth is proposed in which uptake of K(+) and Cl(-) ions in concert with IAA-induced plasma membrane H(+)-ATPase activity changes the membrane potential to a value needed for turgor adjustment during the growth of maize coleoptile cells.

Multifractal Analysis of the Influence of Indole-3-Acetic Acid on Fast-Activating Vacuolar (FV) Channels of Beta vulgaris L. Taproot Cells.

In this paper, the multifractal properties of the ion current time series in the fast-activating vacuolar (FV) channels of Beta vulgaris L. taproot cells were investigated. These channels are permeable for only monovalent cations and mediate K+ at very low concentrations of cytosolic Ca2+ and large voltages of either polarity. Using the patch clamp technique, the currents of the FV channels in red beet taproot vacuoles were recorded and analysed by using the multifractal detrended fluctuation analysis (MFDFA) method. The activity of the FV channels depended on the external potential and was sensitive to the auxin. It was also shown that the singularity spectrum of the ion current in the FV channels is non-singular, and the multifractal parameters, i.e., the generalised Hurst exponent and the singularity spectrum, were modified in the presence of IAA. Taking into account the obtained results, it can be suggested that the multifractal properties of fast-activating vacuolar (FV) K+ channels, indicating the existence of long-term memory, should be taken into account in the molecular mechanism of the auxin-induced growth of plant cells.

Effect of K+ and Ca2+ on the indole-3-acetic acid- and fusicoccin-induced growth and membrane potential in maize coleoptile cells.

The role of potassium (K(+)) and calcium (Ca(2+)) in the regulation of plant growth and development is complex and needs a diverse range of physiological studies. Both elements are essential for satisfactory crop production. Here, the effects of K(+) and Ca(2+) ions on endogenous growth and growth in the presence of either indole-3-acetic acid (IAA) or fusicoccin (FC) were studied in maize (Zea mays) coleoptiles. Membrane potentials of coleoptile parenchymal cells, incubated in media containing IAA, FC and different concentrations of K(+) and Ca(2+), were also determined. Growth experiments have shown that in the absence of K(+) in the incubation medium, both endogenous and IAA- or FC-induced growth were significantly inhibited by 0.1 and 1 mM Ca(2+), respectively, while in the presence of 1 mM K(+) they were inhibited only by 1 mM Ca(2+). At 10 mM K(+), endogenous growth and growth induced by either IAA or FC did not depend on Ca(2+) concentration. TEA-Cl, a potassium channel blocker, added 1 h before IAA or FC, caused a reduction of growth by 59 or 45 %, respectively. In contrast to TEA-Cl, verapamil, the Ca(2+) channel blocker, did not affect IAA- and FC-induced growth. It was also found that in parenchymal cells of maize coleoptile segments, membrane potential (Em) was strongly affected by the medium K(+), independently of Ca(2+). However, lack of Ca(2+) in the incubation medium significantly reduced the IAA- and FC-induced membrane potential hyperpolarization. TEA-Cl applied to the control medium in the same way as in growth experiments caused Em hyperpolarization synergistic with hyperpolarization produced by IAA or FC. Verapamil did not change either the Em of parenchymal cells incubated in the control medium or the IAA- and FC-induced membrane hyperpolarization. The data presented here have been discussed considering the role of K(+) uptake channels in regulation of plant cell growth.

Effect of Trimethyltin Chloride on Slow Vacuolar (SV) Channels in Vacuoles from Red Beet (Beta vulgaris L.) Taproots.

In the present study, patch-clamp techniques have been used to investigate the effect of trimethyltin chloride (Met3SnCl) on the slow vacuolar (SV) channels in vacuoles from red beet (Beta vulgaris L.) taproots. Activity of SV channels has been measured in whole-vacuole and cytosolic side-out patch configurations. It was found that addition of trimethyltin chloride to the bath solution suppressed, in a concentration-dependent manner, SV currents in red beet vacuoles. The time constant, τ, increased significantly in the presence of the organotin. When single channel activity was analyzed, only little channel activity could be recorded at 100 µM Met3SnCl. Trimethyltin chloride added to the bath medium significantly decreased (by ca. threefold at 100 µM Met3SnCl and at 100 mV voltage, as compared to the control medium) the open probability of single channels. Single channel recordings obtained in the presence and absence of trimethyltin chloride showed that the organotin only slightly (by <10%) decreased the unitary conductance of single channels. It was also found that Met3SnCl significantly diminished the number of SV channel openings, whereas it did not change the opening times of the channels. Taking into account the above and the fact that under the here applied experimental conditions (pH = 7.5) Met3SnCl is a non-dissociated (more lipophilic) compound, we suggest that the suppression of SV currents observed in the presence of the organotin results probably from its hydrophobic properties allowing this compound to translocate near the selectivity filter of the channel.

[Allergy to formaldehyde]. / Uczulenie na parapaste.

The article presents a case of 56-year-old patient, female, who developed generalized urticaria following dental treatment. The wheals appeared four hours after she left the clinic. During stomatological treatment she was given parapaste, a complex drug, consisting of cinchocaine, eugenol, paraformaldehyde and glycerin. She was admitted to hospital and was treated with prednison and cetirizine. The symptoms resolved in one day and she was discharged. This was the second episode of urticarial reaction to after dental visit. In a previous case, several months earlier, she received parapaste and Scandonest, local anaesthetic. She visited an allergologist after three weeks and skin prick tests with common allergens were done. All were negative. Then the prick tests with the ingredients of parapaste were done resulting in 5 mm diameter wheal to the solution of formaldehyde. We performed RAST to formaldehyde - the result was confirming the presence of antibodies to formaldehyde in a third class. The diagnosis of sensitization to formaldehyde was made. The incidence of allergy to dental drugs was discussed.

[Reasons for hospitalization in patients with chronic obstructive pulmonary disease(COPD)]. / Przyczyny hospitalizacji chorych na przewlekla obturacyjna chorobe pluc (POChP).

Reasons for cohort of patients with COPD hospitalisation in a 90-bed internal diseases ward within the 1st January and 31st December 2001 are discussed. Four hundred ten medical histories of COPD patients were analysed. The number of hospitalizations and the number of patients were distinguished. The majority of findings have been related to the number of hospitalizations. Percentage of COPD patients, spirometries and history of smoking have been related to the number of patients. In 12 months, the total number of hospitalizations has been 3,352, including 452 hospitalizations of COPD patients--13.8%. 2,731 patients, including 333 COPD (12.2%) patients, have been hospitalised once or more times. Analysed group included 1,975 female (58.9%) and 1,377 male patients (41.1%), and 225 female (49.8%) and 227 male patients (50.2%) in the COPD group. Average age was 64.8 +/- 16.3 years (men--61.7 +/- 16.4 and women--67.0 +/- 15.8), while average age of COPD patients was 68.8 +/- 10.4 years (men--68.6 +/- 10.2 and women--69.0 +/- 10.7). Average hospitalisation duration was 9.0 +/- 5.7 days, while for COPD patients--10.5 +/- 6.0 days. In this cohort COPD has been the main diagnosis in the 195 hospitalizations. Severity of the disease has been graded according to GOLD and the Polish Phtisiopneumonological Society recommendations. A structure of deaths in COPD group, treatment and history of smoking were analysed.