Quantification of heterocyclic aromatic amines in fried meat by HPTLC/UV-FLD and HPLC/UV-FLD: a comparison of two methods

A recently developed HPTLC/UV-FLD method was compared to the routinely used HPLC/UV-FLD method for the quantification of heterocyclic aromatic amines (HAA) formed at trace levels during the heating process of meat. For formation of these process contaminants under normal cooking conditions, beef patties were fried in a double-contact grill at 230 degrees C for five different frying times and extracted by solid-phase extraction. The HAAs most frequently found, that is, 2-amino-1-methyl-6-phenylimidazo[4,5- b]pyridine (PhIP), 2-amino-3,8-dimethylimidazo[4,5- f]quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5- f]quinoxaline (4,8-DiMeIQx), 9 H-pyrido[3,4- b]indole (norharman), and 1-methyl-9 H-pyrido[3,4- b]indole (harman), were quantified by two chromatographic methods, which were orthogonal to each other (normal versus reversed phase system). Both methods showed a similar performance and good correlation of the results ( R (2) between 0.8875 and 0.9751). The comparison of running costs and run time in routine analysis proved HPTLC/UV-FLD to be more economical (factor of 3) and faster (factor of 4) due to its capability of parallel chromatography. The HAA findings calculated by standard addition increased with the heating time from <1 to 33 microg/kg related to 3-6 min of frying time. The precision (RSD) was between 7 and 49% (HPTLC) and between 5 and 38% (HPLC) at these very low HAA levels formed.     

Nitrogen fertilization does not affect micronutrient uptake in grain maize (Zea mays L.)

Abstract

Due to continuous single nitrogen fertilization, we hypothesized a built-up deficiency of micronutrients in crops that would limit plant growth and crop quality. In 2-year field experiments using urea-N fertilized grain maize (Zea mays L.), hybrid KWS 2376 at 0, 120 and 240 kg N ha^?1 crop uptake of Zn, Mn, Cu and Fe was studied at DC 32, DC 61 and in the grain harvested. Micronutrient contents at DC 32 stage – 1st node (aboveground phytomass) and DC 61 – flowering (ear leaf) were all at levels indicative of adequate micronutrient supply to the crop. At both sampling occasions the Fe:Zn and Fe:Mn ratios were adequate implying that Fe did not inhibit the uptake of Zn and Mn. The application of nitrogen increased the Fe content at the 1st sampling in both years; in the second year the same was also the case for the Zn content. Nitrogen nutrition increased the contents of Mn and Fe at the 2nd sampling only in year 2; in the other treatments no changes were observed in the micronutrient contents. Micronutrient correlations in the grain were discovered between Zn and Mn contents and between Fe and Mn contents. In the second year the highest N-rate significantly increased the Fe and Zn content of the grain compared with the lower rates of nitrogen fertilization. Grain yields were not affected by the rate of nitrogen and ranged between 13.65 and 14.34 t ha^?1 (1st year) and between 13.68 and 14.18 t ha^?1 (2nd year). Nitrogen fertilization did not reduce the content of micronutrients in the plant or grain of maize. It is evident that the continuous single use of N fertilization so far has not resulted in a micronutrient deficiency of the plants limiting the nutrient density of the grain or reducing its quality.     

The influence of progesterone and prostaglandin F2α on decorin and the adhesion of caruncular epithelial cells of bovine placenta at early-mid pregnancy—Part II

One of the most important processes determining the proper course of gestation and its physiological termination in cows is the adhesion of epithelial cells allowing for direct contact of maternal and foetal parts of the placenta. Throughout pregnancy, placental cells are under strict hormonal control, which among others regulates the concentration and activity of specific proteins participating in the extracellular matrix remodelling of foetal membranes. The aim of the study was to evaluate the influence of progesterone and prostaglandin F_2α on the adhesion of epithelial cells at early-mid pregnancy in cows. Additionally, the impact of selected hormones on anti-adhesive properties of decorin was evaluated. Caruncular epithelial cells were isolated from healthy cows during pregnancy, immediately after slaughter. Primary cell cultures derived from the 2nd and 4th month of gestation were used in the experiments. The viability of cells was assessed by MTT assay. The adhesion of cells to fibronectin was measured spectrophotometrically. The activity of metalloproteinases was confirmed by the metalloproteinase assay. Progesterone (10^–5 and 10^–7 mol/L) and prostaglandin F_2α (10^–4_, 10^–5 and 10^–7 mol/L) increased the viability of bovine caruncular epithelial cells in the 2nd month of pregnancy. The treatment with prostaglandin F_2α significantly reduced the number of adherent cells from the 2nd month of gestation at the doses of 10^–4 and 10^–5 mol/L. Both progesterone and prostaglandin F_2α were shown to have an effect of decorin resulting in both a decrease in metalloproteinase activity and an increase in adhesion of cells to fibronectin.     

The premetazoan ancestry of cadherins

Cadherin-mediated cell adhesion and signaling is essential for metazoan development and yet is absent from all other multicellular organisms. We found cadherin genes at numbers similar to those observed in complex metazoans in one of the closest single-celled relatives of metazoans, the choanoflagellate Monosiga brevicollis. Because the evolution of metazoans from a single-celled ancestor required novel cell adhesion and signaling mechanisms, the discovery of diverse cadherins in choanoflagellates suggests that cadherins may have contributed to metazoan origins.     

Mercury content and its bioconcentration factors in wild mushrooms at Łukta and Morag,northeastern Poland

Total concentrations of mercury were determined using cold-vapor atomic absorption spectroscopy (CV-AAS) in the fruiting bodies of 16 species of wild mushrooms and underlying soil (0-10 cm) substrates collected in the areas of the Communes of Morag and ?ukta in the county of Ostróda in northeastern Poland in 1997-1998. A total of 174 composite samples of caps, 174 stalks, 80 whole fruiting bodies (collectively 1254 specimens), and 252 soils were examined. Among several species of mushrooms analyzed, the greatest concentrations were between 1300 and 71000 ng.g(-1) of dry matter. These levels were found in the caps of Sweating mushroom (Clitocybe rivulosa), King Bolete (Boletus edulis), and Common Puffball (Lycoperdon perlatum) and also were characterized by the highest bioconcentration factors (BCF) for Hg, which ranged between 160 +/- 82 and 110 +/- 34. The cap to stalk quotient for mercury concentrations was approximately 2 for most of the species except Poison Pax (Paxilus involutus), which had a greater concentration in caps than in stalks and a quotient of 4.4 +/- 7.2. Hg concentrations in the underlying soil substrates (0-10 cm layer) ranged between 21 +/- 21 and 390 +/- 130 ng.g(-1) of dry matter. The results showed that the consumption of mushrooms, considered to be the sole dietary source of mercury at the highest or mean element concentrations found, is not hazardous at daily ingestion rates of less than 70 and 210 g of fresh product, which would result in a hazard index value of less than unity.     

Effect of fast protein liquid chromatography fractionated salivary gland extracts from different ixodid tick species on interleukin-8 binding to its cell receptors

Interleukin-8 plays a critical role in inflammatory processes. Hence generation of molecules with anti-IL-8 activity is likely to be important for successful feeding and for survival of the ticks. Anti-IL-8 activity was studied in saliva of three ixodid tick species--Dermacentor reticulatus (Fabricius, 1794), Rhipicephalus appendiculatus Neumann, 1901, and Amblyomma variegatum (Fabricius, 1794). The greatest activity was shown in saliva prepared from D. reticulatus. The activity was attributed to tick salivary gland molecules that bind to IL-8, preventing binding of the chemokine to its specific receptor, rather than to occupation of the IL-8 cell receptor by the tick molecules. The distribution of anti-IL-8 activity in fast protein liquid chromatography (FPLC) fractions of salivary gland extracts (SGE) derived from adult female D. reticulatus, R. appendiculatus and A. variegatum was compared directly by both ELISA and receptor-binding inhibition assays. The correspondence in results with fractions of SGE from ELISA is consistent with detection of tick molecules that inhibit IL-8 binding to its receptor. As IL-8 is an important chemoattractant and activator of neutrophils, the presence of an anti-IL-8 activity in tick saliva indicates that neutrophils play an important role in the host response to parasitism by ticks.     

Differenzierung des Pansenepithels des Rindes während der intrauterinen Entwicklung

The prenatal development of the bovine ruminal epithelium was studied with light- and electronmicroscopical techniques. During the period of the nonstratified epithelium a pseudostratified epithelium is found in the dorso-cranial part, whereas the other areas possess a one-layered epithelium, which is, like the pseudostratified epithelium, transformed to a multilayered epithelium from the 7th week onwards. From the 9th week the period of the stratified epithelium starts with the formation of the stratum profundum and stratum superficiale. First signs of keratinization are seen in the superficial cells from 2.3 months onwards. With 4 months fetal cornified cells can be identified, with 5.5 months a single-layered stratum basale is seen on the differentiating papillar connective tissue, and the superficial cells are transformed to balloon-cells. In suprapapillar areas, a stratum spinosum is formed at the prenatal age of 7.5 months. During epitheliogenesis a horizontal and vertical differentiation of the cells can be observed. The first one includes the differentiation of undifferentiated, embryonal cells to the basal cells of the stratum profundum, the latter the development of the basal cells to spinous cells and then to fetal cornified and balloon-cells. The ultrastructural changes during the process of keratinization were especially considered.     

LH-β and FSH-β mRNA expression in nesting and post-breeding three-spined stickleback, Gasterosteus aculeatus, and effects of castration on expression of LH-β, FSH-β and spiggin mRNA

The mRNA expression of the LH- and FSH- subunits were measured in nesting and post-breeding male three-spined sticklebacks, Gasterosteus aculetaus, as well as in castrated and sham-operated nesting males. Furthermore, expression of an androgen induced kidney protein, spiggin, and 11-ketotestosterone (11KT) levels, were measured in the castrated and sham-operated males. Nesting males had significantly higher levels of both LH- and FSH- mRNA expression compared to post-breeding males. Furthermore, sham-operated males had significantly higher levels of LH- mRNA and spiggin mRNA expression than the castrated fish. Expression of FSH-, on the other hand, did not differ between castrated and sham-operated males. There were strong positive individual correlations between circulating levels of 11KT on the one hand and expressions of LH- and spiggin mRNA, whereas the correlation between 11KT levels and FSH- mRNA was weak. The negative effect of castration on -LH mRNA indicates that gonadal hormones stimulate this expression, whereas this was not the case for -FSH. The observed decline in -LH expression after the end of the breeding season may be the result of cessation of the gonadal stimulation of the pituitary. On the other hand, it is not likely that this can explain the decline in FSH- expression.