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.

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.

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.

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).

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.

[Development of an engraftable skin equivalent based on matriderm with human keratinocytes and fibroblasts]. / Entwicklung eines transplantierbaren Hautäquivalentes auf Basis von Matriderm mit menschlichen Keratinozyten und Fibroblasten.

A cell-based wound coverage with keratinocytes and fibroblasts on the basis of a commercially available dermal substitute (Matriderm ((R)), Kollagen/Elastin matrix) was generated, in order to treat wide burn wounds. First the expansion of keratinocytes was optimised and the culturing time was minimised. Raw material was 1-2 cm (2) split skin. Dermis and epidermis were separated by enzymatic treatment with thermolysin. After treatment of both compartments with trypsin and collagenase I, keratinocytes and fibroblasts were isolated and expanded in collagen I coated dishes. After 10 days fibroblasts were seeded on Matriderm ((R)). After cultivation of the fibroblasts-containing matrix for one week keratinocytes were seeded on top. After an additional week of submersed cultivation the matrix was lifted up to the air-liquid interface to initiate epidermal cell differentiation. After 16 days in the air-liquid interphase the matrix was fixed and underwent immunohistochemical and electron microscopic analysis. Histological analysis showed a regularly stratification of the epidermal part. We observed collagen IV, a marker for the basement membrane, between epidermis and dermis. Desmoglein and the differentiation markers involucrine and cytokeratin 10 were found in the suprabasal layers of the epidermis. Electron microscopic analysis showed the basement membrane in the epidermal junction zone as well as cell-cell connections in the form of desmosomes. Late differentiation characteristics, like granular structures and the cornified layer, were found in the stratum granulosum and stratum corneum. Our results demonstrate that a skin equivalent can be generated by using a collagen/elastin matrix, with an expansion rate of 50-100-fold. This skin equivalent may be useful for covering deep wounds.

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.

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.

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.

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.

Moniliformin accumulation in kernels of oats used for food and feed purposes.

Inoculation experiments with 12 genotypes of oats (9 lines: CHD 894, CHD 1095, CHD 1296, CHD 1607, CHD 1653, CHD 1692, STH 2293, STH 2393, STH 2494, STH 2694 and 2 cultivars: Farys, Slawko) were performed during the vegetation seasons of 1999 - 2000 in Zamosc region in the South - Eastern part of Poland.Panicles of oats were inoculated with a conidial suspension ofFusarium avenaceum, which caused the following average reductions of traits: yield by 37.8 % and weight of 1000 kernels by 40.2 % respectively.The highest susceptibility to scab development caused by theF. avenaceum inoculation was found particularly for two lines (CHD 894, and CHD 1296) while cultivar Farys and line CHD 1607 exhibited the highest resistance to the disease in terms of TKW and yield reductions after inoculation. During 2 consecutive years (1999 and 2000 respectively) the average level of moniliformin accumulation (mg/kg) in positive samples was respectively 0.51 and 0.12 the highest being 1.11 and 0.26, while the lowest was 0.30 and 0.01.

Fusariotoxins in kernels of winter wheat cultivars field samples collected during 1993 in Poland.

In South-Eastern region of Poland (near Lublin), where frequency of scab (fusariosis) is much higher than in other parts of the country, during harvest of 1993 kernels of 25 winter wheat cultivars were collected. On the basis of morphological studies Fusarium graminearum was found in 42% of investigated samples while other fungi appeared less frequently: F. nivale and F. poae (35%), F. avenaceum (31%) and F. culmorum (12%). Chemical analysis (by HPLC) revealed that the tested cultivars were contaminated with deoxynivalenol (96% of investigated samples), its acetyl derivatives (48%), nivalenol (76%) and moniliformin (28%). The average levels of the metabolite concentrations were as follows: 104; 16; 97; and 63 micrograms/kg, respectively. Co-occurrence of 2 toxic metabolites was found in the following percentage of the positive samples: deoxynivalenol and nivalenol (72%), deoxynivalenol and moniliformin, as well as nivalenol and moniliformin (24%). Usually (71-83% of contaminated samples) mycotoxins were accumulated in the concentration range > or = 10, < 100 micrograms/kg.

X trisomy in an infertile bitch: cytogenetic, anatomic, and histologic studies.

Three copies of the X chromosome were identified in a 5-year-old mixed breed infertile bitch. One year after the cytogenetic examination, the bitch died due to gastritis hemorrhagica, an inflammation of the mucus coat of the stomach. Dental studies showed congenital lack of some premolar and molar teeth. Ovaries were of normal shape and size. Also, histologic sections of the ovaries revealed their normal structure, with two corpora lutea and primary follicles. Phenotypic effects of X trisomy are discussed.

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.

Deoxynivalenol accumulation and other scab symptoms in six romanian wheat genotypes inoculated withFusarium graminearum.

At anthesis, under field conditions at Fundulea, each of 6 Romanian winter wheat genotypes was inoculated with 3Fusarium graminearum isolates used individually.Fusarium head blight (FHB) was assessed according to the following traits: relative weight of spikes (RWS), percentage of Fusarium damaged kernels (FDK), relative weight of kernels per head (RWKH), area under the disease progress curve (AUDPC) and deoxynivalenol (DON) content in total sample of kernels. Correlations between these traits and parameters revealed important differences between examined wheat genotypes in: DON accumulation, progress of FHB development, yield reduction, and models of host - pathogen interactions in theTriticum - Fusarium pathosystem. Significant correlations between different attributes of FHB were found forFusarium isolate 1 which is a moderate producer of DON (0.89 µg g(-1)). Weight of spike was significantly correlated with weight of kernels per spike (r = 0.93**) and with percentage of damaged kernels (r = - 0.87**), while FDK was highly correlated with RWKH (r = - 0.85*) and with DON content (r = 0.82*). Area under the disease progress curve was also found to be significantly correlated with DON content (r = 0.86*).

Ochratoxin A in porcine blood and in consumed feed samples.

The aim of the study was to estimate occurrence of ochratoxin A (OA) in feeds and the metabolite residues in porcine blood serum in Poland. Samples were collected in the period from February to May, 1999, in the southern Wielkopolska region. Altogether 40 and 45 samples of feed and porcine blood serum, respectively, were analyzed for OA. Percentage of samples contaminated with OA, both in case of feeds and blood, collected in the winter season was considerably higher than that for the spring season. The percentages for feeds were as follows: 47.6 and 26.3 %, while for porcine serum: 66.7 and 50.0 %, respectively winter and spring. In 25 % of cases ochratoxin A was present in both types of investigated material (feed, blood), whereas in 27.5 % of samples this metabolite was detected in blood only, or in 7.5 % only in the feed. The presence of OA was found neither in the feed nor in the serum in 40 % of all cases. In subgroups (feed, blood) the concentration in the whole collective of positive samples were in the range 0.3-13.5 ng/g and 0.3-69.5 ng/ml, respectively, while median values were 2.3 ng/g and 6.0 ng/ml. Only one feed and three porcine serum samples, were found to be contaminated at concentration levels higher than 10 ng/g or 10 ng/ml.