Agaricus bisporus compost improves the potential of Salix purpurea × viminalis hybrid for copper accumulation.

The aim of the study was to determine the ability of spent mushroom compost (SMC) from the production of Agaricus bisporus (A. bisporus) to stimulate the growth and efficiency of copper (Cu) accumulation by Salix purpurea × viminalis hybrid. Roots, shoots and leaves were analysed in terms of total Cu content and selected biometric parameters. Due to the absence of information regarding the physiological response of the studied plant, low molecular weight organic acids (LMWOAs), phenolic compounds and salicylic acid (SA) contents were investigated. The obtained results clearly demonstrate the effectiveness (usefulness) of SMC in promoting the growth and stimulation of Cu accumulation by the studied Salix taxon. The highest Cu content in roots and shoots was found at the 10% SMC addition (507±22 and 380±11 mg kg(-1) DW, respectively), while there was a reduction of the content in leaves and young shoots (109±8 and 124±7 mg kg(-1) DW, respectively). In terms of physiological response, lowered secretion of LMWOAs, biosynthesis of phenolic compounds and SA, as well as accumulation of soluble sugars in Salix leaves was observed with SMC addition. Simultaneously, an elevation of the total phenolic content in leaves of plants cultivated with SMC was observed, considered as antioxidant biomolecules.

Salicylic acid - a potential biomarker of tobacco Bel-W3 cell death developed as a response to ground level ozone under ambient conditions.

Salicylic acid content and benzoic acid 2-hydroxylase (BA2H) activity were investigated in tobacco Bel-W3 and Bel-B leaves after exposure to tropospheric ozone in the conditions of ambient air. Plants were exposed in accordance with a standard methodology for ozone biomonitoring, in a three-year experiment. Free salicylic acid (SA), conjugated with glucose (SAG), and as a product of the BA2H activity was quantified with HPLC. In order to evaluate ozone injuries of leaves, an open source image analysis software was employed. Plants exposure to ambient ozone resulted in enhanced BA2H activity and intensified salicylic acid biosynthesis in leaves of Bel-W3 cultivar showing visible ozone injuries. The BA2H activity significantly correlated with SAG for ozone-exposed Bel-W3 plants. Both injuries and salicylic acid biosynthesis rate depended on the growth phase of leaves and nearly linear correlation between SA content and injuries was found for particular leaves of Bel-W3.

Oral mucosa model based on a collagen-elastin matrix.

BACKGROUND AND OBJECTIVE: The collagen-elastin matrix (Matriderm(®)) is used to treat deep and full-thickness burns and was recently described as a suitable scaffold for tissue engineering. The aim of the present study was to investigate the biocompatibility of Matriderm(®) for gingival use through creation of an oral mucosa model ex vivo. MATERIAL AND METHODS: Gingival fibroblasts and keratinocytes were cultured. A dermal area on the base of the collagen-elastin matrix was repopulated with fibroblasts. After 14 days, keratinocytes were seeded on this dermal area to engineer a multilayered mucosa. Analysis of the architecture was performed using light and electron microscopy. Immunohistochemical detection of collagen IV and cytokeratin was carried out. RESULTS: Based on this scaffold we generated a multilayered oral mucosa-like structure. Histological, immunohistochemical and electron microscopic analysis of the dermal/epidermal junction showed a typical basement membrane and hemidesmosomal structures. Neighboring keratinocytes formed desmosomes in the epidermal sections. Cytokeratin was detectable in all epidermal layers. These experiments revealed that the collagen-elastin matrix was highly biocompatible with gingival cells under ex vivo conditions. CONCLUSION: Employing tissue-engineering techniques with dermal and epidermal cells from the gingiva, a multilayered oral mucosa was generated and characterized with respect to biocompatibility for Matriderm(®). The results indicate that Matriderm(®) is suitable for the ex vivo growth of gingival tissue cells and is a useful scaffold with possible applications in periodontal therapy.

Influence of Ca/Mg ratio on phytoextraction properties of Salix viminalis. II. Secretion of low molecular weight organic acids to the rhizosphere.

A hydroponic experiment in a phytotron was performed to investigate the effect of two different Ca/Mg ratios (4:1 and 1:10) and trace element ions (Cd, Cu, Pb and Zn) in solution on the efficiency of low molecular weight organic acid (LMWOA) formation in Salix viminalis rhizosphere. Depending on the Ca/Mg ratio and presence of selected trace elements at 0.5mM concentration, the amount and kind of LMWOAs in the rhizosphere were significantly affected. In physiological 4:1 Ca/Mg ratio the following complex of acids was observed: malonic (Pb, Zn), citric, lactic, maleic and succinic (Zn) acids. Under 1:10 Ca/Mg ratio, citric (Cd, Zn), maleic and succinic (Cd, Cu, Pb, Zn) acids were seen. Additionally, high accumulation of zinc and copper in all systems was observed, with the exception of those where one of the metals was at higher concentration. Summing up, the results indicate a significant role of LMWOAs in Salix phytoremediation abilities. Both effects can be modulated depending on the mutual Ca/Mg ratio.

Cadmium and lead uptake by Salix viminalis under modified Ca/Mg ratio.

The studies were focused on the influence of mutual changes in the ratio of calcium and magnesium in soil on the efficiency of cadmium and lead uptake and accumulation by Salix viminalis. In the environmental experiment the soil was modified according to the natural Ca/Mg ratio with the experimental groups 4:1 (physiological ratio), 20:1, and 1:10, and was characterized by the amounts of the most important elements (including heavy metals) and selected soil parameters. Efficiency of lead and cadmium accumulation was in the order of Ca/Mg ratios 1:10 > 4:1 > 20:1, and was higher in the shoots at 0.5 m height when compared to 0.1 m above the ground. Under Cd and Pb supplementation the level of both metals was 2-fold higher, while bioaccumulation factor values according to selected literature indicated weak accumulation. When the seasonal metal uptake was analysed (from April to October), the highest accumulation efficiency was observed in the first two periods (April to August) and was Ca/Mg ratio independent. Salix growth was restrained under 1:10 Ca/Mg ratio while it was stimulated under 20:1 ratio, which was opposite to the metal sorption. Summing up, increased calcium level in soil (20:1 Ca/Mg) resulted in limited sorption of heavy metals and stimulated biomass productivity. Under increased magnesium concentration (1:10 Ca/Mg) in relation to the natural 4:1 ratio the accumulation efficiency was the greatest but plant growth was inhibited.

Mycotoxins and mycotoxicoses under climatic conditions of Poland.

Ochratoxin A (OA), zearalenone (ZON), moniliformin (MON) as well as trichotecenes and fumonisines (FUM) are naturally occurring contaminants of cereals and animal feed. They pose a health risk not only to humans but also to livestock and, as a consequence, may cause economical losses either due to unfavorable effects on domestic animals themselves or to an increased potential for health effects in human beings consuming mycotoxin-contaminated edible animal products. At present, large-scale studies are carried out in EU countries to determine a safe, admissible concentration of these toxins in cereals and their processed products. The aim of this review is to collect and summarize information concerning the properties, occurrence and toxicity of these mycotoxins.

Accumulation of phosphorylated I kappaB alpha and activated IKK in nodes of Ranvier.

AIMS: Nuclear factor-kappaB (NF-kappaB) is an ubiquitously expressed transcription factor that modulates inducible gene transcription crucial for the regulation of immunity, inflammatory processes, and cell survival. In the mammalian nervous system, constitutive NF-kappaB activation is considered to promote neuronal cell survival by preventing apoptosis. Increasing evidence suggests a critical role for NF-kappaB activation in acute and chronic neurodegenerative diseases. Recently, a striking enrichment of phosphorylated I kappaB alpha (pI kappaB alpha) and activated I KappaB Kinase (IKK), two key components of the NF-kappaB activation pathway, was demonstrated in the axon initial segment (AIS) of neurons. As the AIS shares fundamental features with nodes of Ranvier (NR), we examined whether pI kappaB alpha and activated IKK are also enriched in NR. METHODS: Double-immunofluorescence labelling was performed with vibratome sections of the rodent central and peripheral nervous system. Sections were analysed using confocal laser scanning microscopy and preembedding electron microscopy. RESULTS: Here we report a remarkable accumulation of pI kappaB alpha and activated IKK in NR in the central and peripheral nervous system. Immunolabelling for both proteins extended from NR into the adjacent paranode. pI kappaB alpha predominantly accumulated within the cytoplasm and was associated with fasciculated microtubules. This association was confirmed by electron microscopy. By comparison, activated IKK preferentially clustered beneath the cytoplasmic membrane. CONCLUSION: In conclusion, the coincident accumulation of pI kappaB alpha and activated IKK in AIS and NR suggests that these specific axonal compartments contribute to neuronal NF-kappaB activation.

Factors determining the sward utilisation of winter pasture in cattle feeding.

In this review article, factors determining the sward utilisation of winter pasture in cattle feeding were defined and analyzed. The results from literature and own investigations have shown that yield and quality of autumn-saved herbage on winter pasture are determined by pratotechnical measures such as harvest date in winter and pre-utilisation date in summer, botanical composition of sward, and particularly the dominated grass species or community, nitrogen fertilisation and weather conditions in winter. The date of winter harvest as a dominating factor affects the development of dry matter yield as well as the digestibility of organic matter and nutrients concentration in herbage during winter. For the management of winter grazing systems it could be obtained that crude protein and energy concentration of the tested autumn-saved herbage met the requirements of suckler cows or beef cattle until the end of the year if they were pre-utilised in July. The weather conditions during autumn-winter period are important factor determining the accumulation of secondary metabolites formed by field fungi in herbage of winter pasture.

Different calcium storage pools in vascular smooth muscle cells from spontaneously hypertensive and normotensive Wistar-Kyoto rats.

OBJECTIVE: To evaluate whether the distribution of intracellular free calcium may be impaired in primary hypertension. DESIGN: Cytosolic free calcium and stored calcium were investigated in cultured vascular smooth muscle cells from spontaneously hypertensive rats (SHR). METHODS: The concentrations of intracellular and stored calcium were investigated in cultured vascular smooth muscle cells from spontaneously hypertensive rats aged 6 months from the Münster strain (SHR) and from age-matched normotensive Wistar-Kyoto (WKY) rats. Vascular smooth muscle cells were grown on coverslips, and fluorescence measurements of the intracellular calcium concentration were performed using fura-2. The different effects of thapsigargin, a selective Ca-ATPase inhibitor, and of angiotensin II (Ang II) on the calcium storage pools were investigated. RESULTS: In the absence of external calcium thapsigargin produced a dose-dependent transient increase in the concentration of intracellular calcium in vascular smooth muscle cells. The thapsigargin-induced maximum peak increase in the concentration of intracellular calcium was not significantly different in SHR and WKY rats. After depletion of the thapsigargin-sensitive calcium pools the addition of 100 nmol/l Ang II produced a rise in the concentration of intracellular calcium in vascular smooth muscle cells from SHR and WKY rats. Using vascular smooth muscle cells from the SHR the Ang II-induced increase in the concentration of intracellular calcium was not significantly different in the presence and absence of thapsigargin, indicating that the calcium pools depleted by thapsigargin and Ang II do not overlap significantly in vascular smooth muscle cells from SHR. In contrast, in the WKY rats the response to Ang II was significantly diminished after depletion of the thapsigargin-sensitive pool. When Ang II and thapsigargin were administered in the reverse order, i.e. Ang II before thapsigargin, the thapsigargin response was diminished in the WKY rats but not in the SHR. CONCLUSION: SHR differ from WKY rats in having vascular smooth muscle cells that contain thapsigargin-sensitive calcium storage pools that are distinct from the Ang II-sensitive calcium pools.

Effect of Na,K-ATPase inhibition on cytosolic free calcium ions in vascular smooth muscle cells of spontaneously hypertensive and normotensive rats.

OBJECTIVE: To investigate the role of Na(+)-Ca2+ exchange in the regulation of cytosolic free Ca2+ and the pathogenesis of primary hypertension. METHOD: Cytosolic free Ca2+ ([Ca2+]i) in cultured vascular smooth muscle cells from normotensive and spontaneously hypertensive rats of the Münster strain was measured using the fluorescent dye fura-2 after inhibition of Na+,K+ATPase by ouabain and after addition of angiotensin II. RESULTS: [Ca2+]i showed a rapid increase together with a depolarization of membrane potential as measured by merocyanine 540. The ouabain-induced increase in [Ca2+]i was blocked in Ca(2+)-free medium and by nifedipine, but incubation with the inhibitor of the Na(+)-Ca2+ exchange, NiCl2, did not diminish the effect of ouabain. Likewise, in Na(+)-free medium the response to ouabain was not suppressed. The angiotensin II-induced changes in [Ca2+]i were diminished in Ca(2+)-free medium and by nifedipine, but enhanced by NiCl2. CONCLUSION: The increase in [Ca2+]i after Na+,K+ ATPase inhibition is not due to a modulation of Na(+)-Ca2+ exchange, but to a Ca2+ influx through Ca2+ channels. Changes in Na(+)-Ca2+ exchange caused by Na+,K+ ATPase inhibition may not play an important role in vascular smooth muscle cells of spontaneously hypertensive rats.

Genetic determination of variability of barley doubled haploids inoculated with Fusarium culmorum (W.G.Sm.) Sacc. with regard to mycotoxin accumulation and reduction in yield traits.

The genetic determination of variability of barley doubled haploid (DH) lines in regard of their susceptibility to Fusarium head blight caused by Fusarium culmorum was studied. The susceptibility was evaluated in 3-year field experiment on the basis of reduction in yield traits and myotoxin accumulation in infected kernels. The following traits were analysed in inoculated and control plants: kernel number and weight per ear, 1000-kernel weight, percentage of plump kernels (>2.5 mm), deoxynivalenol (DON) content and nivalenol (NIV) content of kernels. On the basis of the obtained data, heritability coefficient (ratio of genotypic to phenotypic variance) was assessed, and genetic parameters as well as the number of effective factors were estimated. Heritability coefficients calculated from two-way analysis of variance, i.e.regarding the influence of years and year x genotype interaction, appeared to be exceptionally low and ranged from 5.2% for the reduction in plump kernels to 38.2% for the reduction in 1000-kernel weight. In the case of mycotoxin accumulation about 60% of the observed variability in NIV concentrations and 30% in DON concentration resulted form genetic differences among lines. Additive effects of genes were important for all the analysed traits. Significant effects of dominance and dominance x dominance were observed for 1000-kernel weight and percentage of plump kernels. Moreover, it was found that the observed variability in yield trait reduction resulted from segregation of 5-6 effective factors, DON contents from 4 factors, while NIV content from 5 factors.

Dietary origin of mycotoxins with estrogenic potential and possible health implications to female dogs.

In Poland, occurrence of toxigenic fungi in cereals, foods, feeds and their components as well as mycotoxins accumulation in such material has been studied by numerous teams including our research group for over thirty years since 1969. Mostly cereal kernels and feeds have been examined for presence of toxigenic fungi, their toxigenic potential and natural contamination with mycotoxins. Ochratoxin A, deoxynivalenol, nivalenol and moniliformin were found to be significant contaminants of agricultural products in high percentage of cereal grain samples. The profile of toxic metabolites was similar but the concentration levels of the toxins were lower when compared to already published data of the same climate zone. Zearalenone (ZEA), a nonsteroidal mycotoxin with estrogen-like activity, is synthesized by molds (Fusarium) commonly contaminating poorly stored agricultural products and foodstuffs. Since in the course of examinations and during surgical procedures performed in dogs, ovarian cysts were detected and because frequently this is the first stage of the endometrica pyometra complex (EPC) found in approximately 30% of the females we assume that both factors, mycotoxins (ZEA) and pathological aberrations are possibly related in these animals. Similar activity of the toxin (possibly present in pelleted dog feed) and effects including infertility of female dogs is speculated with indication and suggestion on necessity of additional studies on the problem.

Accumulation of mycotoxins in forage of winter pasture during prolonged utilisation of sward.

The concentration of mycotoxins in sward depending on time of pre-utilisation in summer and date of harvest in winter was analysed during the 2000-2002 vegetation seasons. Additionally, the yield of pasture sward was estimated. Higher concentrations of ochratoxin A--respectively: 0.55 and 050 ng/g DM--were found in the pasture sward harvested in December and January when compared to the sward collected in November (0.36 ng/g DM). The average concentration of zearalenone varied from 2.74 ng/g DM in December, to 6.00 ng/g DM in November and 9.77 ng/g DM in January. The highest concentration of ochratoxin A was 1.82 ng/g DM and zearalenone 47.89 ng/g DM. The highest number of samples in which ochratoxin A exceeded the level of 0.3 ng/g DM was noted in sward harvested in January (61.1%), while in the case of zearalenone, the number of samples containing this mycotoxin at a level exceeding 3.0 ng/g DM varied from 55.6% in December to 66.7% in November and January. It seems that the percentage of ochratoxin A positive samples and the toxin concentration level in the sward of winter pastures increased during the time of sward regrowth.

[A test for determining daily doses of consumption of ochratoxin A by humans in Poland]. / Próba okreslenia dawki dziennego spozycia ochratoksyny A przez czlowieka w polsce.

A trial was undertaken for determination of the average daily amount of ochratoxin A consumed with contaminated food by adults in Poland. Contamination of food with ochratoxin A was estimated to be 0.448 ng/g, and the mean level of this metabolite in human serum was 0.27 ng/cm3. These values showed a considerable similarity when they were analysed by simplified conversion factors of Mortensen et al.

Kinetics of fumonisin B1 formation in maize ears inoculated with Fusarium verticillioides.

The kinetics of fumonisin B1 (FB1) biosynthesis have been examined in ears of four botanical varieties Zea mays var. indentata, Zea mays var. indurata, Zea mays var. saccharata and Zea mays var. everta inoculated with F. verticillioides isolates at silking stage. The level of mycotoxin accumulated in kernels was correlated with sample harvest time (r = 0.73) and ergosterol content (r = 0.70). FB1 biosynthesis was influenced by amylose, starch and moisture contents, which undergo dynamic changes during grain formation. A comparative analysis of ear infection rates and efficiency of FB1 biosynthesis showed that these are separate characters and their expression could be independent.

Reaction of winter wheat (Triticum aestivum L.) cultivars to infection with Fusarium spp.: mycotoxin contamination in grain and chaff.

This study compares the susceptibility of winter wheat (Triticum aestivum L.) cultivars to Fusarium head blight (FHB) and accumulation of mycotoxins in kernels and chaff under different climatic conditions in two locations-Cerekwica near Poznan (Central West Poland) and Sitaniec, near Zamosc, Lublin region (South East Poland). Very high variations were found in the concentrations of mycotoxins (zearalenone, ZEA; nivalenol, NIV; deoxynivalenol, DON; moniliformin, MON) in examined fractions: Fusarium-damaged kernels (FDK) and healthy looking kernels (HLK) and in chaff for individual cultivars in both locations. In most cases, significantly higher concentrations of investigated toxins were recorded in wheat from the area of Lublin than from Poznan (p < 0.05). The highest Fusarium infection rates and mycotoxin biosynthesis levels were observed in the Lublin location, with the percentage of the FDK fraction ranging 8.1-81.6. In this region, ZEA concentration (microg g(-1)) after inoculation with F. culmorum and F. graminearum ranged from 0.02-0.48 and 0.32-1.04, respectively. In the Poznan area, the toxin concentrations were considerably lower, ranging 0.01-0.10 and 0.03-0.13 microg g(-1) for both isolates, respectively. The concentration of DON was significantly higher than ZEA or NIV levels. The levels of MON accumulation (microg g(-1)) in the FDK fraction were between 0.14 and 1.73 (Poznan area) and ND (not detected) to 2.51 (Lublin area). F. avenaceum infection rate ranged 7-35% in samples where the toxin was detected.

Formation of fumonisins and other secondary metabolites by Fusarium oxysporum and F. proliferatum: a comparative study.

The principal aim of this study was to estimate the formation of fumonisins (FB(1) and FB(2)), moniliformin (MON), and ergosterol (ERG) by Fusarium oxysporum and Fusarium proliferatum, while the formation of beauvericin (BEA) was estimated by the latter Fusarium species only. Moreover, the effect of temperature on the biosynthesis of mycotoxins was also evaluated. Fumonisins were formed by F. proliferatum, with the highest yield at 18 degrees C (720.0-1976.6 microg g(-1) for FB(1), 74.2-670.8 microg g(-1) for FB(2)) and only by three of four F. oxysporum strains at a very low level (0.02-4.77 microg g(-1) for FB(1), 0.02-2.15 microg g(-1) for FB(2)). The amount of MON formed by F. proliferatum was the highest (p < 0.001) at 32 degrees C (3056.87 microg g(-1)), while MON biosynthesis by F. oxysporum was lower 227.54 microg g(-1) (p < 0.001). BEA was produced by F. proliferatum with the highest level at 25 degrees C (p < 0.001). ERG-recognized as an indicator of fungal biomass development and as a consequence of mycotoxin formation-was found at the highest concentration at a biosynthesis temperature of 25 degrees C for F. proliferatum and F. oxysporum (p < 0.001).