Band 66, Heft 4

The ter operon, which is found on a large conjugative plasmid, pTE53, from the uropathogenic strain Escherichia coli KL53, mediates heavy metal ion resistance to tellurium compounds. Here we present the function of the terW gene, which is transcribed from its own promoter in the opposite direction to the terZABCDE loci and encodes a protein of 155 amino acids. A TerW protein containing an N-terminal His-tag was overexpressed, purified and analysed for the in vitro DNA-binding. Green fluorescent protein fusions, a fluorimetric assay, nonradioactive gel retardation and capillary electrophoresis footprinting were used to examine the interaction between the bioinformatically-predicted DNA-binding region and TerW. Here we show that TerW binds specifically to the potential promoter region of the terZABCDE genes, which are responsible for the tellurite resistance of the Escherichia coli host cells. Our results suggest that the transcriptional regulation of ter genes is operon-like.

Denaturing gradient gel electrophoresis (DGGE) and multivariate statistical analytical methods were applied to investigate the spatial variation of bacterial community structure in the Pearl River estuary sediment and to address the relationship between microbial community composition and bottom water chemistry in ten different stations. Preliminary results of sequencing analysis of the excised DGGE bands suggested that α-Proteobacteria, γ-Proteobacteria, Acidobacteria and Actinobacteria were the dominant bacterial groups in the Pearl River estuary sediment. Results of multidimensional scaling analysis of these field data suggested that the composition of bacterial communities varied with sampling sites. Finally, canonical correspondence analysis of the data of environmental variables and bacterial community suggested that bacterial community structure was significantly influenced by the change of environmental variables (total phosphorus, nitrite, ammonium, dissolved oxygen, pH and salinity).

Following traditional morphological concepts, the genus Coronastrum is considered to be a rare member of the Scenedesmaceae (Chorophyceae). This classification may be called into question when molecular data are taken into account as well. Recent molecular phylogenetic studies revealed the polyphyletic origin of the family Scenedesmaceae within the Chlorophyceae and Trebouxiophyceae. In a combined approach of morphological analyses, SSU/ITS rRNA gene phylogeny and comparison of the ITS secondary structure, we analysed the systematics of Coronastrum strains available in public strain collections. Our molecular analyses revealed a new subclade within the Chlorella clade of the Chlorellaceae consisting of Coronastrum ellipsoideum, two strains with Dictyosphaerium-like morphology and one strain which fits the description of the genus Parachlorella. Four additional strains formed together a new lineage within the genus Parachlorella in the Parachlorella clade of the Chlorellaceae. These strains differ from the already known Parachlorella species in complementary base changes within the ITS2 and are here described for the first time as Parachlorella hussii sp. nov.

The hamate dandelion section (Taraxacum sect. Hamata H. Øllg.) represents a morphologically tight group of taxa, distributed in the oceanic and sub-oceanic regions of Europe. In Central Europe, it is mostly confined to freshly moist meadows, as well as places in the shade within urban areas. The known distribution area in Europe until now extended towards the E part of the Czech Republic and S Poland. During the past six years, we have discovered an additional eleven localities of hamate dandelions in NW and N Slovakia, which represents a corrected south-eastern limit for the distribution area of this section. In this paper, we discuss seven recognized taxa (T. boekmanii, T. fusciflorum, T. hamatiforme, Taraxacum hamatum, T. lamprophyllum, T. pseudohamatum, and T. quadrans). We provide a determinative key to the Slovakian hamate dandelions, brief species descriptions, comments on their distribution, a distribution map of the section in Slovakia, as well as images of all of the newly found species.

Morphological analysis of the water lily (Nymphaea) was carried out to verify distribution of two closely related species Nymphaea alba and N. candida in the large swampy area in Eastern Poland, Polesie Zachodnie. A total of 25 lakes were investigated across the region. Data were collected from 20 lakes of different origin. Based on morphological characters of flowers and leaves the specimens varied continuously from “pure” N. candida to N. alba with high number of plants with mixed morphology, commonly known as N. × borealis. The intermediate morphotypes were common in all over the study area, whereas plants with exact morphology of N. candida were found mostly in humic lakes, in the eastern part of the region. The results provide evidences that N. candida occurs beyond formerly recognized southern border of its range in the country.

The flower of Rhododendron luteum (L.) Sweet has a pentamerous structure with radial symmetry. The anthers filament surface is covered by dense non-glandular hairs to the half of the height. The tubular anther dehisces along creating two openings in the anther-sac walls and the viscous pollen is released through two splits along the anther lobes. The pistil is pentamerous and the axial channel is filled with a mucilaginous secretion product which is continuous with the exudate on the stigma surface. The stigmatic papillae are densely packed and their exudate is stained intensively red for carbohydrates, while pollen grains are stained positively for lipids. The five-locular ovary has isomerous carpels (syncarpous gynoecium) and the ovary surface is covered by numerous, densely-packed glandular and non-glandular hairs protecting the nectar against transpiration. Numerous ovules per locule occur with one integument and a thin-walled megasporiangium. In carpels, oil cells occur sporadically as solitary idioblasts, located around the vascular bundles. Transmitting tissue cells contain a large central, electron translucent vacuole, filling most of the cell containing dark osmiophilic bodies homogenous or granular in appearance.

Gentiana cruciata L. (cross gentian) is a medicinal and ornamental plant. The root extracts of this species are known to exhibit many curative properties. The natural Gentiana populations are exposed to great danger because of their uncontrolled usage. In this study, hairy roots from Gentiana cruciata L. stem and leaf explants belonging to three different clones were induced by inoculation with four different Agrobacterium rhizogenes wild strains namely A4, 15834, 8196 and R1000. Induction of the root transformation was significantly dependent on the explant type used. On the other hand, the genotype and bacterial strain had no significant effect on hairy root formation. Hairy root formation percentages of the explants varied between 5.6–33.3% in the stem explants, and between 0.0–6.7% in the leaf explants. Transformations of the hairy roots were confirmed by PCR using rolC specific primers, and revealed the absence of contaminating A. rhizogenes with virC primers. Total of twelve hairy root clones were obtained, and their secondary metabolite content was also analyzed by HPLC. Quantitative results exhibited that gentiopicroside was the most abundant compound in all root samples. Furthermore, metabolites such as loganic acid, swertiamarin, and sweroside were also identified and quantified in the samples.

The vegetation of the aquatic and marshland habitats of the Orava region (north of Slovakia) was studied in 2009, using traditional phytosociological methods. Sixteen aquatic and eighteen marsh plant communities were described within 96 phytosociological relevés by using TWINSPAN with the application of the dominance principle. Three associations, Potametum alpini, Potametum zizii and Ranunculo-Juncetum bulbosi, were found as new communities for Slovakia. Myriophylletum verticillati, Potametum nodosi, Potametum graminei and Alisma gramineum community were recorded in the northernmost localities in Slovakia. Among marsh communities, Calletum palustris is very rare, both in the Orava region and in Slovakia as a whole. According to Ellenberg’s indicator values (EIV), moisture was evaluated as the main ecological gradient. Plant communities are ordered along the first Detrended Correspodence Analysis (DCA) axis in a typical hydroseries (Potametea → Lemnetea → Phragmition communis → Phalaridion arundinaceae, Oenanthion aquaticae and Sparganio-Glycerion → Magnocaricion elatae). The second DCA axis was most correlated with EIV for nutrients. Among the five directly measured ecological characteristics (temperature, pH and conductivity of water, water depth, and substrate type), conductivity of water (0.44, P < 0.01) and substrate type (0.32, P < 0.05) were the statistically significant variables explaining the variability along the first DCA axis.

Invasions of Typha (cattail) and/or Phragmites (common reed) in wetland ecosystems result in changes in species richness, diversity and composition of vascular plants. These invasions are particularly harmful in lakes where threatened species and/or communities are found. The spread of two species of Typha (T. angustifolia and T. latifolia) and of Phragmites australis in the Stagnone Lake, on Capraia Island (Tuscan Archipelago — Mediterranean sea) was studied. We report this progressive invasion, documented by means of a series of vegetation maps (1991, 1995, 1998 and 2009). The expansion rate of the three invasive helophytes and the shrinking of the aquatic communities were studied using a GIS system. The impact of the spread of these three species on the floristic characteristics of the plant communities and the lake vegetation in general, was analysed by means of 15 plots of 1 m2 in 2000 and in 2009. Statistical analysis of the two series shows a significant change in the floristic composition of the communities as a result of the invasion process. Many important groups of species, such as many aquatic species, decrease in number and in cover value.

During research on gymnamoebae in freshwater ponds near the Modra town (Slovakia), in one sample of water and sediment of the 71 samples examined a fungal infection of Amoebophilus simplex Barron, 1983 (Zygomycota: Zoopagales) in Mayorella vespertilioides Page, 1983 (Amoebozoa: Flabellinea) was detected for the first time. From 27 observed amoebae, 9 specimens (33%) were infected. Populations of other species recorded in the sample, Mayorella penardi and Korotnevella stella, were uninfected.

It is known that the microturbellarian Stenostomum sphagnetorum, a common ciliate predator, is very sensitive against different types of toxins produced by other ciliates for chemical defense, and consequently, it is not able to capture and ingests these ciliates. In particular, when the predator tries to attack one of these toxin-secreting ciliates, it is forced to regurgitate the captured prey in response to the toxin discharged from the ciliates. In this study it is shown that after repeated attacks by S. sphagnetorum against two species of toxic ciliates (Spirostomum ambiguum and Spirostomum teres), the predator acquires a behavior of prey selection that leads to the exclusion of these ciliates among the possible prey and to the distinction between edible and inedible (toxic) ciliates. This learned behavior, which is maintained for days, is lost only after the asexual reproduction of the microturbellarian. In addition, S. sphagnetorum learns to recognize and avoid specimens of S. ambiguum and S. teres artificially deprived of their toxins, strongly suggesting that the toxins are not exclusively associated in the prey recognition of the microturbellarian.

For the first time, Piscicola brylinskae was described from Lake Vechten in the village of Bunnik, near Utrecht — The Netherlands. Until now, P. brylinskae has been found in Poland in Lake Maróz and in the Łyna River near Olsztyn (the northern part of Warmian-Masurian voivodeship). Thanks to proper conservation the coloration of P. brylinskae was described for the first time. Applying 32 indexes resulted that P. brylinskae clustered to Caspiobdella fadejewi. Analysis based on 113 non-metric characteristics has shown that P. brylinskae is most similar to Piscicola margaritae. Though, P. brylinskae do not form with P. margaritae dichotomic branching nor with any other species from this cluster, it could confirm that it is an individual species of the Piscicola genus.

The rotifer community of the canyon shaped Slapy Reservoir was studied over four years, from 2000 to 2003. A total of 38 rotifer taxa were identified in the pelagial zone. The dominant species, Keratella cochlearis, K. quadrata, Polyarthra dolichoptera, P. vulgaris, Synchaeta lakowitziana and Pompholyx sulcata, were similar to other reservoirs in the same area. Tychoplanktic species, which are typical for communities of upstream reservoirs, are reduced in the Slapy Reservoir. The seasonal fluctuation of rotifer abundance is characterized by a significant spring peak, clear water depression, summer peak and sometimes a lower peak in autumn. Fluctuation of the rotifer species shows a highly unified trend through the years with very low variation through the years. It results from the input of hypolimnetic water from an upstream reservoir, which erased a large part of the effect of year to year climatic fluctuations.

The hypothesis of associating pseudoscorpions with bird nest types was tested on the basis of an analysis of 480 specimens. Eleven pseudoscorpion species were found in 171 nests of 28 different bird species collected in Slovakia, Austria and the Czech Republic. The frequent appearance of Cheiridium museorum, Dactylochelifer latreillii, Chernes hahnii, Dendrochernes cyrneus and Allochernes wideri was confirmed. High proportion and association of Pselaphochernes scorpioides in hoopoe hollow nests with decomposed substrate, D. cyrneus in the Eurasian tree sparrow nest boxes and A. wideri in the nests of the tawny owls, the European scops owls and the European roller was proved. In contrast, C. hahnii and D. latreillii were related to the nest fauna of blackbirds and song thrushes, C. museorum to the nests of white wagtails situated on the ground and on buildings and C. cancroides to the nests in synanthropic habitats. Until present, the occurrence of 22 pseudoscorpion species has been confirmed in the bird nests of Central Europe based on the obtained results and published resources. According to the association to bird nests these pseudoscorpions were classified as (1) nidixenous species — Chthonius fuscimanus, C. tetrachelatus, Mundochthonius styriacus, Neobisium carcinoides, N. crassifemoratum, N. inaequale, N. sylvaticum, Chernes cimicoides, C. similis, C. vicinus, Allochernes powelli, Lamprochernes chyzeri, L. nodosus and Larca lata; and (2) nidiphilous species — C. museorum, C. cancroides, A. wideri, D. cyrneus, D. latreillii, C. hahnii, D. panzeri and P. scorpioides.

In spring 2004, the mark-release-recapture study was conducted in the Osijek area, covering the total of 171 km2, to describe dispersal pattern of three floodwater mosquito species (Aedes vexans, Ochlerotatus sticticus and Oc. caspius). Forty CO2 baited Center for Disease Control and Prevention (CDC) traps were set at distances varying from 0.95 to 16.7 km from release site. Fifty thousand mosquitoes were released, and a total of 22 marked mosquitoes were recaptured in 12 traps, 82% of which were collected in the first six days after release. The maximum distance of recaptured mosquito (Oc. sticticus) was found at 11.68 km from the release site. During the study, the average dispersal rate per day for mosquitoes ranged from 0.96 km to 3.6 km in regard to different species.

Bombyx mori parvo-like virus is a small, icosahedral virus containing two single-stranded linear DNA molecules (VD1, VD2). To date, little is known about this virus, how to package. VD2-ORF1 encoded a large viral structural protein with predicted molecular mass of 133 kDa, and this protein was named as P133. It is unusual for 20–22 nm icosahedral viruses to posses so large a structural protein and the function of this protein is still unknown. In this study, the transcription of P133 was examined with quantitative real-time PCR, and the results demonstrated that the mRNA of P133 could be detected from 28 h post inoculation and kept increasing until 72 h post inoculation. P133 C-terminus (P133C) and P133 N-terminus (P133N) were cloned and expressed in E. coli BL21; then the resulting polypeptides were used to produce antibody, respectively. Western blot analysis showed that the protein in virions recognized by anti-P133C and anti-P133N antibody had the same molecular weight, indicating that VD2-ORF1 encoded a viral structural protein without leaky scanning.

Roost microclimate plays an important role in the survival, growth and reproduction in microbats. Entering torpor is one of the main energy saving mechanisms commonly used by microbats. The use of torpor is affected by roost microclimate and seasonally differs between the two sexes in relation to their reproductive condition. Consequently, thermal properties of male and female roosts should differ. To test this hypothesis, we compared temperature parameters of two anthropogenic day roosts of Daubenton’s bats with a different structure of the population inhabiting them. In accordance with our predictions, the roost occupied by a male-dominated colony was colder and more fluctuant than the maternity roost with a female-dominated population. However, using of the two roosts changed during the season in response to changing energetic demands of the two sexes. While males were almost absent in the warmer maternity roost during pregnancy and lactation, they appeared in this roost during the post-lactation when mating starts. In contrast, females did not use the colder (male) roost until the time of weaning of juveniles, i.e., the time when their thermoregulatory needs change and they may benefit from using colder roost. Our study provides the evidence that the same roost may be used by individuals of different sex and reproductive state in different periods of the year. Generalizations about roost selection without knowledge of temporal variation in roost use and microclimatic conditions should be taken with caution. Anthropogenic roosts may be advantageous to Daubenton’s bats as these can provide a variety of suitable microclimates and/or more space for roosting than tree cavities.

The aim of our study was to evaluate the parasite — host interactions at apoptosis level. We studied histopathological changes and time course of apoptosis in the duodenum during Eimeria acervulina infection. One-day-old broiler chicks were randomly allocated into two equal groups. At the age of two weeks the first group was experimentally infected with a pure suspension of sporulated E. acervulina oocysts. The second group served as a negative control. Tissue samples from the upper part of duodenum were obtained at 0.5, 1, 2, 3, 4, 5 and 6 days post infection. Biopsies of duodenum were studied immunohistochemically using DeadEnd™ Colometric TUNEL System for apoptosis detection in duodenal mucosa. Number of parasites in duodenal epithelium was also investigated. Our experimental results demonstrate: (i) macroscopic and histopathological changes in epithelium detected mainly in proximal segment of duodenum in infected groups; (ii) the number of developmental stages of E. acervulina (DSEA) during our trial increased, reaching the maximum 5 days post infection (dpi) (332.2 ± 16.12) (mean ± SEM), whereas the amount of DSEA declined significantly as late as 6 dpi (124.6 ± 3.91); (iii) the highest apoptosis level was recorded in initiatory 0.5 dpi (13.2 ± 1.02) and on the end of parasite development cycle after 5 dpi (12.6 ± 1.36). Finally, results showed that there was a period of inhibition of apoptosis during infection by E. acervulina.

Ionizing radiation commonly used in the radiotherapy of brain tumours can cause adverse side effects to surrounding normal brain tissue. The most significant response of adult brain to radiation damage is induction of apoptosis. The adult mammalian subventricular zone (SVZ) of the brain lateral ventricles (LV) and their subsequent lateral ventricular extension, the rostral migratory stream (RMS), is one of the few areas, which retains the ability to generate new neurons and glial cells throughout life. Taking into account the fact, that ionizing radiation is one of the strongest exogenous factors affecting cell proliferation, the aim of the present study was to investigate the occurrence of radiation-induced apoptosis in this neurogenic region. Adult male Wistar rats were investigated 1, 5 or 10 days after single whole-body gamma irradiation with the dose of 3 Gy. Apoptotic cell death was determined by in situ labelling of DNA nick ends (TUNEL) and fluorescence microscopy evaluation of TUNEL-positive cells. Considerable increase of apoptotic TUNEL-positive cells was observed 24 hrs after irradiation in caudal parts of RMS; i.e. in the vertical arm and elbow of RMS. Initial increase was followed by strong reduction of apoptosis in the RMS and by secondary over-accumulation of apoptotic cells in the animals that survived ten days after exposure. Results showed, that the proliferating population of cells, arisen in SVZ are highly sensitive to radiation-induced apoptosis. This observation should have implications for clinical radiotherapy to avoid complications in therapeutic brain irradiation.

Stem cells play pivotal role in the development of tissues and organs, as well as they maintain homeostasis and integrity of multicellular organisms. Human dental pulp-derived stem cells are capable of differentiation into osteoblasts, odontoblasts, adipocytes and neuronal-like cells and can have potential use in tissue regeneration. The aim of this study was a detailed description of human dental pulp-derived stem cells (HDPSCs) cultivated in vitro ultrastructural morphology. HDPSCs were isolated from the dental pulp of impacted third molars from healthy donors. Transmission electron microscopic analysis showed typical ultrastructural morphology similar to mesenchymal stem cells from other organs. The morphology of HDPSCs cells reflects their proteosynthetic and metabolic activity. Each cell contained spherical or irregularly-shaped large pale nucleus with a large amount of euchromatine. Nuclei had noticeable nucleolus (or two nucleoli) located nearby the nuclear envelope. Abundant cisterns of rough endoplasmic reticulum and numerous coated matrix vesicles as well as granules of glycogen were present in their cytoplasm. Nearby the nucleus small, elongated mitochondria were placed. Most of HDPSCs created and secreted vesicles; in plasmalemma bounded amorphous electron-lucide granules and also few glycogen granules. These secretory vesicles had around 0.5 μm in diameter. We assume that it can be a special type of communication between cells, probably paracrine type of cell signalling, but its real function is still unknown.

Obesity is a known risk factor for postmenopausal cervical cancer. In human, plasma adiponectin (ADPN) levels are inversely associated with obesity. ADPN may influence the pathogenesis of cervical cancer. In this paper, the effects of ADPN on the proliferation of the cervical cancer HeLa cells and its underlying mechanism were studied. We discovered that two ADPN receptors were expressed in HeLa cells and ADPN inhibited the proliferation of HeLa cells. Western-blotting showed that ADPN activated AMP kinase in HeLa cells. Reverse transcriptase-polymerase chain reaction revealed ADPN down-regulated the expression of cell cycle regulators cyclin D1 and c-myc, and induced the expression of p21WAF1/CIP1 and p53. These results indicate that ADPN inhibits proliferation and induces apoptosis of HeLa cells by altering the expression of cell cycle regulators.

Angiogenesis is involved in tumor growth and metastasis. Endostatin inhibits angiogenesis, but its precise mechanism is not fully understood. To clarify signal transduction involved in endostatin-induced angiogenesis inhibition (endothelial cell growth inhibition), it is important to identify an endostatin receptor, which is the aim of the present study. We hypothesized that vascular endothelial cadherin (VE-cadherin) is an endostatin receptor and found that endostatin induced apoptosis in cultured calf pulmonary artery endothelium (CPAE) cells. Immunoprecipitation and western blots revealed that endostatin specifically bound to VE-cadherin in a Ca2+-dependent manner. Binding of endostatin to VE-cadherin induced tyrosine phosphorylation of VE-cadherin, β-catenin recruitment, and endothelial cell death. Antisense oligonucleotides against VE-cadherin rescued endostatin-induced endothelial cell death. Inhibition of tyrosine phosphorylation of VE-cadherin inhibited endostatin-induced β-catenin recruitment and CPAE cell death. Taken together, we conclude that VE-cadherin is an endostatin receptor.

Epidemiological studies indicate that 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, or statins, play a role in inhibition of several human neoplasia including breast cancer. In this study, chemopreventive effects of atorvastatin in N-methyl-N-nitrosourea-induced mammary carcinogenesis in female rats were evaluated. Atorvastatin was administered in the diet at two concentrations: 10 mg/kg (ATOR 10) and 100 mg/kg (ATOR 100). Atorvastatin treatment began 8 days prior to carcinogen administration and subsequently continued for 15 weeks till the end of the experiment. Atorvastatin at a higher dose suppressed tumor frequency by 80.5% (P = 0.0008) and tumor incidence by 49.5% (P = 0.015), and extended latency period by 14 days (P = 0.076) when compared to the control group. Atorvastatin at a lower dose did not significantly alter tumor parameters in comparison with the control group. In the specimens of mammary tumors, atorvastatin (in the ATOR 100 group) significantly decreased mRNA expression of Bcl-2 gene but non-significantly increased Bax mRNA expression compared to control group. Atorvastatin administration did not alter serum concentration of triacylglycerols, total cholesterol, and LDL cholesterol in comparison with controls. This study is the first report on tumor suppressive effect of atorvastatin in rat mammary carcinogenesis.