Beata Ostachowicz

Establishment of arbuscular mycorrhizal plants originating from xerothermic grasslands on heavy metal rich industrial wastes–new solution for waste revegetation

Industrial waste substrata, rich in heavy metals, are poorly suited for plant growth. Efforts are made to establish an appropriate plant cover to reduce erosion and further contamination. Grasses are the usual solution, as they grow fast, thrive on poor substrata and have well-developed root systems. Some of them are also highly dependent on mycorrhizal symbiosis that supports their growth especially on poor and polluted soils. However, the commercially available grasses often meet a lack of well established mycorrhiza on the site and the introduced plant populations dramatically decrease with time, despite large financial input including covering the substratum with soil and intensive watering. The aim of this paper was to select proper plants together with mycorrhizal fungi that could accelerate the establishment of the vegetation and improve its diversity under these extreme conditions, minimizing the financial costs of the reclamation (no use of soil layering and watering). The experiments were carried out under field and laboratory conditions. The plant seeds used originated from dry calcareous grasslands. The seeds were germinated under field conditions or in pots filled with soil supplemented with substratum from the industrial wastes. The seedlings were inoculated with AM fungi and introduced on the field plots a few weeks after germination. The inoculum consisted of either crude inoculum harvested from the dry calcareous grasslands or strains originating from polluted areas. Plants colonized by mycorrhizal fungi established well in the experimental plots. The results suggest that inocula from dry calcareous grasslands are potentially useful in revegetation of industrial wastes. Although in several cases the photosynthetic activity of plants was lower than at the natural sites, almost all plants survived and formed seeds. In all experiments the plant vitality was estimated on the basis of chlorophyll a fluorescence and was useful to show differences between waste substrata, inocula and coexisting plant species. The interactions between mycorrhizal and non-mycorrhizal plants were studied under greenhouse conditions and at least no negative effect of this coexistence was found.

GPR39 (zinc receptor) knockout mice exhibit depression-like behavior and CREB/BDNF down-regulation in the hippocampus.

Background: Zinc may act as a neurotransmitter in the central nervous system by activation of the GPR39 metabotropic receptors. Methods: In the present study, we investigated whether GPR39 knockout would cause depressive-like and/or anxiety-like behavior, as measured by the forced swim test, tail suspension test, and light/dark test. We also investigated whether lack of GPR39 would change levels of cAMP response element-binding protein (CREB),brain-derived neurotrophic factor (BDNF) and tropomyosin related kinase B (TrkB) protein in the hippocampus and frontal cortex of GPR39 knockout mice subjected to the forced swim test, as measured by Western-blot analysis. Results: In this study, GPR39 knockout mice showed an increased immobility time in both the forced swim test and tail suspension test, indicating depressive-like behavior and displayed anxiety-like phenotype. GPR39 knockout mice had lower CREB and BDNF levels in the hippocampus, but not in the frontal cortex, which indicates region specificity for the impaired CREB/BDNF pathway (which is important in antidepressant response) in the absence of GPR39. There were no changes in TrkB protein in either structure. In the present study, we also investigated activity in the hypothalamus-pituitary-adrenal axis under both zinc- and GPR39-deficient conditions. Zinc-deficient mice had higher serum corticosterone levels and lower glucocorticoid receptor levels in the hippocampus and frontal cortex. Conclusions: There were no changes in the GPR39 knockout mice in comparison with the wild-type control mice, which does not support a role of GPR39 in hypothalamus-pituitary-adrenal axis regulation. The results of this study indicate the involvement of the GPR39 Zn2+-sensing receptor in the pathophysiology of depression with component of anxiety.

The involvement of the GPR39-Zn(2+)-sensing receptor in the pathophysiology of depression. Studies in rodent models and suicide victims.

Zinc is one of the most important trace elements in our body. Patients suffering from depression show lower serum zinc levels compared to healthy controls. Zincs antagonism to the glutamatergic system seems to be responsible for mood recovery. Recent years have shown that zinc may regulate neurotransmission via the metabotropic GPR39 receptor. Activation of the GPR39-Zn(2+)-sensing receptor (GPR39) triggers diverse neuronal pathways leading to a cAMP-responsive element binding the protein (CREB) expression, which then induces synthesis of the brain-derived neurotrophic factor and, in turn, activation of the Tropomyosin receptor kinase B (TrkB) receptor. In the present study, we investigated the alteration of the GPR39 in different models of depression, such as zinc deficiency and olfactory bulbectomy and in suicide victims. Additionaly, we focused on CREB-BDNF/TrkB under zinc deficient conditions in mice. To demonstrate depressive-like behaviour, a standard and modified forced swim test (FST) was performed. To evaluate expression of GPR39, CREB, BDNF and TrkB, Western Blot analysis was used. Zinc deficient mice and rats showed decreased GPR39 expression in the hippocampus and frontal cortex. A decreased level of hippocampal and cortical GPR39 was also observed in suicide victims. In contrast, increased GPR39 in the hippocampus of olfactory bulbectomized rats was observed. Additionally, we found a decreased expression of CREB, BDNF and TrkB only in the hippocampus of zinc-deficient mice. Our present study demonstrates the associacion of the GPR39 Zn(2+)-sensing receptor in the pathomechanism of depression. Down-regulation of CREB, BDNF, TrkB and GPR39 receptor found under zinc-deficient conditions in the hippocampus, may play an important role in the pathophysiology of mood disorders, since most of patients suffering from depression show lower serum zinc.

Zinc deficiency in rats is associated with up-regulation of hippocampal NMDA receptor

RATIONALE Data indicated that zinc deficiency may contribute to the development of depression; however changes induced by zinc deficiency are not fully described. OBJECTIVES In the present paper we tested whether the dietary zinc restriction in rats causes alterations in N-methyl-D-aspartate receptor (NMDAR) subunits in brain regions that are relevant to depression. METHODS Male Sprague Dawley rats were fed a zinc adequate diet (ZnA, 50 mg Zn/kg) or a zinc deficient diet (ZnD, 3 mg Zn/kg) for 4 or 6weeks. Then, the behavior of the rats was examined in the forced swim test, sucrose intake test and social interaction test. Western blot assays were used to study the alterations in NMDAR subunits GluN2A and GluN2B and proteins associated with NMDAR signaling in the hippocampus (Hp) and prefrontal cortex (PFC). RESULTS Following 4 or 6 weeks of zinc restriction, behavioral despair, anhedonia and a reduction of social behavior occurred in rats with concomitant increased expression of GluN2A and GluN2B and decreased expression of the PSD-95, p-CREB and BDNF protein levels in the Hp. The up-regulation of GluN2A protein was also found in the PFC, but only after prolonged (6 weeks) zinc deprivation. CONCLUSIONS The procedure of zinc restriction in rats causes behavioral changes that share some similarities to the pathophysiology of depression. Obtained data indicated that depressive-like behavior induced by zinc deficiency is associated with the changes in NMDAR signaling pathway.

Characterisation of 210Pb dated peat core by various X-ray fluorescence techniques

Abstract Ombrotrophic peat bogs form an independent ecosystem fed by atmospheric precipitation. There is a correlation between depth and time, thus sections of peat taken from various depths can indicate the deposition of several elements from both natural and anthropogenic sources. A peat core of 40 cm in length was taken from a peat bog in southern Poland and cut into 2-cm sections. The peat core was dated using the natural radionuclide 210 Pb. Additionally, pollen analysis was performed in order to corroborate the 210 Pb dating. Eighteen elements and the ash content have been determined in all peat sections. Energy Dispersive X-ray Fluorescence analysis (EDXRF) was applied in this work for simultaneous elemental analysis. The concentration depth profiles for the determined elements and ash content profile were constructed. The results show strong time-dependent variations in the concentrations of the different elements. An interpretation of the data is given and correlation between possible influencing factors is discussed. High concentrations of Pb and Zn indicate their anthropogenic origin.

Metal uptake by xerothermic plants introduced into Zn-Pb industrial wastes.

The dusty surfaces of post-flotation wastes contain high concentrations of toxic compounds and spread widely if appropriate vegetation is not introduced. It has been previously established that effective restoration of such waste areas are best met by xerothermic, mycorrhiza-assisted plants (Turnau et al. Plant and Soil 305:267–280, 2008). The aim of the current study was to improve phytostabilisation practices by gaining insight into the elements uptake in plants after their change of habitat. Total Reflection X-ray Fluorescence (TXRF) was employed to evaluate element concentration in the leaves of 23 plant species growing in the wild and on Zn-Pb waste. Higher levels of heavy metals (Zn, Y, As, Pb, Cu) in plants from tailings were usually accompanied by increased Ca concentration, suggesting a possible role of this element in detoxification mechanisms. Also, when compared to grassland specimens, plants from the tailings, exhibited potassium-deficiency. Thus, K-supplementation of the waste substrata should be considered to improve plant growth. Among all the introduced plants, three grass species (Melica transsilvanica, Bromus inermis, Elymus hispidus) and one legume (Anthylis vulneraria) were the most suitable for phytostabilisation. Heavy metal-accumulating properties of Verbascum thapsus need further investigation.

Antidepressant activity of fluoxetine in the zinc deficiency model in rats involves the NMDA receptor complex

The zinc deficiency animal model of depression has been proposed; however, it has not been validated in a detailed manner. We have recently shown that depression-like behavior induced by dietary zinc restriction is associated with up-regulation of hippocampal N-methyl-d-aspartate receptor (NMDAR). Here we examined the effects of chronic administration of a selective serotonin reuptake inhibitor, fluoxetine (FLX), on behavioral and biochemical alterations (within NMDAR signaling pathway) induced by zinc deficiency. Male Sprague Dawley rats were fed a zinc adequate diet (ZnA, 50mg Zn/kg) or a zinc deficient diet (ZnD, 3mg Zn/kg) for 4 weeks. Then, FLX treatment (10mg/kg, i.p.) begun. Following 2 weeks of FLX administration the behavior of the rats was examined in the forced swim test (FST) and the spontaneous locomotor activity test. Twenty four hours later tissue was harvested. The proteins of NMDAR (GluN1, GluN2A and GluN2B) or AMPAR (GluA1) subunits, p-CREB and BDNF in the hippocampus (Western blot) and serum zinc level (TXRF) were examined. Depression-like behavior induced by ZnD in the FST was sensitive to chronic treatment with FLX. ZnD increased levels of GluN1, GluN2A, GluN2B and decreased pS485-GluA1, p-CREB and BDNF proteins. Administration of FLX counteracted the zinc restriction-induced changes in serum zinc level and hippocampal GluN1, GluN2A, GluN2B and p-CREB but not BDNF or pS845-GluA1 protein levels. This finding adds new evidence to the predictive validity of the proposed zinc deficiency model of depression. Antidepressant-like activity of FLX in the zinc deficiency model is associated with NMDAR complex.

Inventory of regulations and analysis techniques for trace elements in some water supply companies in Eastern and Western Europe

With the ultimate objective in contributing to the improvement of the quality control of drinking water and to facilitate the checking of its compliance with the respective regulations, an inventory of analytical techniques used in water supply companies in Eastern and Western European countries is presented. The existing regulations for drinking water and bottled mineral water quality with respect to trace elements in these countries are compared to regulations of the World Health Organisation. The data obtained can be useful for further harmonisation of the respective regulations in Europe.

Early lifetime zinc supplementation protects zinc-deficient diet-induced alterations

Preclinical and clinical data indicate the involvement of zinc in the pathophysiology and therapy of depression. A relationship between zinc-deficiency and depression symptoms was recently proposed. The present study investigated alterations in spontaneous locomotor activity and zinc concentrations in the serum, hippocampus and frontal cortex; these alterations were induced by subjecting rats to a zinc-deficient diet, prior subjected after birth to zinc-supplemented diet. Body weight was significantly reduced in animals subjected to the four-week zinc-deficient diet compared to those subjected to the zinc-adequate diet. The two-week zinc-deficient diet induced a significant increase in locomotor activity in all measured time periods (5, 30 and 60 min by 44-62%). The four-week zinc-deficient diet did not affect locomotor activity, while the six-week zinc-deficient diet resulted in a 45% increase in the 5 min time period. Serum zinc concentrations were significantly reduced (by 29%) in animals subjected to the four-week zinc-deficient diet but not in those subjected to the two- or six-week zinc-deficient diets. The zinc-deficient diet did not influence the zinc concentration in the examined brain regions regardless of the length. These results indicate that post-birth supplementation with zinc may protect zinc-deficient diet-induced rapid alterations in zinc homeostasis.

A comparison of the performance of a fundamental parameter method for analysis of total reflection X-ray fluorescence spectra and determination of trace elements, versus an empirical quantification procedure

Abstract The performance has been compared of two different quantification methods — namely, the commonly used empirical quantification procedure and a fundamental parameter approach — for determination of the mass fractions of elements in particulate-like sample residues on a quartz reflector measured in the total reflection geometry. In the empirical quantification procedure, the spectrometer system needs to be calibrated with the use of samples containing known concentrations of the elements. On the basis of intensities of the X-ray peaks and the known concentration or mass fraction of an internal standard element, by using relative sensitivities of the spectrometer system the concentrations or mass fractions of the elements are calculated. The fundamental parameter approach does not require any calibration of the spectrometer system to be carried out. However, in order to account for an unknown mass per unit area of a sample and sample nonuniformity, an internal standard element is added. The concentrations/mass fractions of the elements to be determined are calculated during fitting a modelled X-ray spectrum to the measured one. The two quantification methods were applied to determine the mass fractions of elements in the cross-sections of a peat core, biological standard reference materials and to determine the concentrations of elements in samples prepared from an aqueous multi-element standard solution.