Volume 62, Issue 4

Mitochondria are the powerhouse of the cell. They play a vital role in the energy metabolism and regulate calcium flux and apoptosis. The recent resurgence of interest in mitochondrial studies is largely attributed to the recognition that mitochondrial dysfunctions lead to various physiopathological disorders, especially in the development and progression of cancer. Mitochondrial DNA is very susceptible to mutations, which lead to respiratory dysfunction and are implicated in many cancers. Mitochondria serve as the molecular target for a structurally diverse group of pharmacological agents in cancer chemotherapy. Biochemical and biophysical characterization have helped to identify several important differences between mitochondria of normal and disease state. Such unique alterations in mitochondrial structure and function could become promising target for the development of new generation drugs.

Five fresh soil Streptomyces spp. strains were isolated, phylogenetically characterized on the basis of 16S rDNA sequences and analyzed for the presence of restriction modification systems. Three type II site-specific endonucleases were detected and partially purified. Two isolated enzymes were isoschizomers of SacI restriction endonuclease recognizing 5′-GAGCTC-3′ sequence; the third one recognised 5′-CCGCGG-3′ sequence and it was an isoschizomer of SacII. SacII like modification was observed in other two isolates having no detectable restriction activity. The lack of correlation between restriction and modification phenotypes and phylogenetic classification of the isolates indicates efficient gene transfer mechanism in the Streptomyces genus.

Six plasmids carrying a snowdrop lectin (Galanthus nivalis agglutinin, GNA) and one of three selection markers were successfully transferred into two sugarcane cultivars (FN81–745 and Badila) via Agrobacterium-mediated transformation. Agrobacterium strains LBA4404, EHA105 and A281 that harboured a super-binary vector were used for sugarcane transformation. The use of the hygromycin (Hyg) resistance gene (hpt II), phosphinothrincin (PPT) resistance gene (bar) or G418 resistance gene (npt II) as a screenable marker facilitated the initial selection of GNA transgenic sugarcane callus with different efficiencies and helped the rapid segregation of individual transformation events. All the three selective marker genes were controlled by CaMV 35S promoter, while GNA gene was controlled by promoter of RSs-1 (rice sucrose synthase-1) or Ubi (maize ubiquitin). Factors important to successful transformation mediated by Agrobacterium tumefaciens were optimized, which included concentration of A. tumefaciens, medium composition, co-cultivated methods with plant tissue, strain virulence and different selective marker genes. An efficient protocol for sugarcane transformation mediated by A. tumefaciens was established. The GNA gene has been integrated into sugarcane genome as demonstrated by PCR and Southern dot blotting detections. The preliminary results from bioassay demonstrated a significant resistance of the transgenic sugarcane plants to woolly aphid (Ceratovacuna lanigera Zehnther) indicating thus the possibility for obtaining a transgenic sugarcane cultivar with resistance to woolly aphid.

The main goal of this work was to make the cDNA-encoding subunit G2 of soybean glycinin, capable of self-assembly in vitro and rich in methionine residues. Two mutants (pSP65/G4SacG2 and pSP65/G4SacG2HG4) were therefore constructed. The constructed mutants were successfully assembled in vitro into oligomers similar to those occurred in the seed. The successful self-assembly was due to the introduction of Sac fragment of Gy4 (the codons of the first 21 amino acid residues), which reported to be the key element in self-assembly into trimers. The mutant pSP65/G4SacG2HG4 included the acidic chain of Gy4 (HG4), which was previously molecularly modified to have three methionine residues. This mutant will be useful in the efforts to improve the seed quality.

We applied the Cocktail method to a large data set of 4 117 relevés of all Slovak vegetation types with the aim to create formalised definitions of all Slovakian mire plant associations. We defined 21 groups of species with the statistical tendency of joint occurrences in vegetation. These groups differed substantially in their position along the pH/calcium gradient. We further defined 24 plant associations according to presence and/or absence of certain groups and/or strong dominance of some species. Only six traditional plant associations were not possible to be reproduced this way. We applied our formalised definitions to the regional data set of mires from the surrounding of the Vysoké Tatry Mts. Combined with frequency-positive fidelity index this method has led to the classification of the majority of vegetation plots into ten associations. When the vegetation types obtained from Cocktail-based classification and from cluster analysis were compared with respect to measured pH and conductivity in the study region, 82% of pairs differed significantly either in pH or in water conductivity in the former classification and 69% in the latter one.

Types and the number of stomata in the following Arum species: Arum italicum Mill., Arum maculatum var. maculatum L. and Arum maculatum var. immaculatum L., Arum alpinum var. pannonicum Terpo. and Arum alpinum var. intermedium Schur. in three different locations in Zablaće, Normanci and Bilje were investigated. The most prevalent stomata type at both upper and lower epidermis for each Arum species was mostly stomata type paracytic, followed by hexacytic, tetracytic or brachyparacytic as far as locality is concerned. Helicocytic type was more prevalent in Arum alpinum var. pannonicum Terpo. and Arum alpinum var. intermedium Schur. at Bilje. Other stomata types were very rare. A striking regularity in the occurrence of stomata types was not found within a single species. Some stomata types, however, were found either at adaxial or abaxial epidermis or were not present at all. The number of stomata per square mm varied from 25 to 651. A statistically significant difference in the number of stomata per square mm at upper and lower epidermis among Arum species was determined in locations Zablaće and Normanci, whereas no statistically significant differences were found in location Bilje.

Changes in the seasonal rhythm of two plant phytocoenoses in a submountain beech forest during secondary succession were studied. Investigations were done on four monitoring plots with different stand density over the period of four successive years. The rhythm of the associations Dentario bulbiferae-Fagetum and Carici pilosae-Fagetum reflects the course of succession processes running six years after the human impact (cutting) in the ecosystem. Results of the phenological observations of the understorey species with the focus on the changes in flowering and colour spectrum allowed to make the comparisons between both associations in connection with different phyto-climatic conditions and in dependence on time. The most conspicuous changes in the seasonal rhythm and structure of the examined associations were found in conditions of the former clear-cut, currently in succession phase. A clear decrease (56%) in number of taxons with the dominance > 1% in one association towards the end of the 4-year study period was detected here. Simultaneously, a decrease in the number of flowering species was observed, while the relative rate of species being in the vegetative stage increased considerably (from 6 to 67%) over the growing season. The course of flowering of both of the associations missed discernible trends and peaks as well as colour spectra were partially changed during four monitored successive years on the formerly unstocked area.

The European method for mapping lichen diversity (LDV) as an indicator of environmental stress/quality in the Czech Republic was evaluated to determine its applicability in this country. A modified Hawksworth & Rose qualitative scale for determining air pollution levels was used for comparative purposes. The study was undertaken in a partially forested and reasonably polluted region of the Bohemian Karst in central Bohemia during 2003. Measurements obtained by the LDV method yielded values in the range 18–65; measurements obtained by the modified Hawksworth & Rose method indicated zones 4–7, with a high correlation with the LDV results. The LDV method proved to be applicable in the region, but its implementation may be limited by the lack of suitable trees and specific landscape ecology.

Research on the diurnal distribution of physical and chemical parameters within a single macrophyte bed was carried out on the shallow Wielkowiejskie Lake (Poland). A non-parametric statistical analysis was used to compare the water quality features in different parts of a Chara hispida habitat including the middle, both edge (vertical and horizontal) parts of a macrophyte plant, and the open water next to-and above the stonewort stand. The obtained results showed a differentiation in the physical-chemical parameters of the environmental conditions within the Chara hispida stand. The greatest variability was found for dissolved oxygen. Its lowest concentrations were noted in the central part of the macrophyte stand, irrespective of the sampling time. The zooplankton communities within the examined Chara bed were strongly influenced by the concentration of dissolved oxygen. It was also found that two main components of zooplankton communities (rotifers and cladocerans) had a similar trend in their spatial and diurnal distribution within the stonewort stand.

Alpine grasslands in the Southern Carpathian Mts, Romania, harbour an extraordinarily high diversity of plants and invertebrates, including Carpathic endemics. In the past decades, intensive sheep grazing has caused a dramatic decrease in biodiversity and even led to eroded soils at many places in the Carpathians. Because of limited food resources, sheep are increasingly forced to graze on steep slopes, which were formerly not grazed by livestock and are considered as local biodiversity hotspots. We examined species richness, abundance and number of endemic vascular plants and terrestrial gastropods on steep slopes that were either grazed by sheep or ungrazed by livestock in two areas of the Southern Carpathians. On calcareous soils in the Bucegi Mts, a total of 177 vascular plant and 19 gastropod species were recorded. Twelve plant species (6.8%) and three gastropod species (15.8%) were endemic to the Carpathians. Grazed sites had lower plant and gastropod species richness than ungrazed sites. Furthermore, grazed sites harboured fewer gastropod species endemic to the Carpathians than ungrazed sites. On acid soils in the Fagaras Mts, a total of 96 vascular plant and nine gastropod species were found. In this mountain area, however, grazed and ungrazed sites did not differ in species richness, abundance and number of endemic plant and gastropod species. Our findings confirm the high biodiversity of grasslands on steep slopes in the Southern Carpathian Mts and caution against increasing grazing pressure in these refuges for relic plants and gastropods as well as for other invertebrates.

A total of 57 stonefly species have been recorded in the Hron River. The natural gradient (slope and stream width) and pollution gradient of the river were defined using CCA based on physical, chemical and stonefly data. Stonefly metrics (abundance, richness/diversity, sensitivy/tolerance and functional metrics) were used to estimate the quality of the Hron River and the degree of proximity to its natural state. Similar results were obtained using two different methods. The first method was based on the homogeneity of variance and the interquartile range of different groups of stretches of the Hron River and the second was based on deviations from the expected values of biological metrics in a given stretch of the river. These values continuously decreased with increasing distance from the spring area, with the exception of the saprobic index, which increased in a downstream direction, and the stonefly average score, which did not change significantly along the whole river flow. The Stonefly Average Score (SAS) metric is universal for a variety of habitats such as the Hron River upstream and downstream, and is a reliable indicator of water quality and the natural course of a stream.

The incubation period of eggs, duration of three nymphal instars, adult longevity and the daily egg-deposition rate of the ischnoceran Phthiraptera, Brueelia amandavae, were determined by rearing the louse in vitro (35 ± 1°C, 75–82% RH, feather diet). The data obtained were utilized for life table construction and determination of the intrinsic rate of natural increase (0.031 per day) and the doubling time (23.45 days) of the louse population. The doubling time of the louse in in vivo experiments was 21.5 days.

The classification methodology based on morphometric data and supervised artificial neural networks (ANN) was tested on five fly species of the parasitoid genera Tachina and Ectophasia (Diptera, Tachinidae). Objects were initially photographed, then digitalized; consequently the picture was scaled and measured by means of an image analyser. The 16 variables used for classification included length of different wing veins or their parts and width of antennal segments. The sex was found to have some influence on the data and was included in the study as another input variable. Better and reliable classification was obtained when data from both the right and left wings were entered, the data from one wing were however found to be sufficient. The prediction success (correct identification of unknown test samples) varied from 88 to 100% throughout the study depending especially on the number of specimens in the training set. Classification of the studied Diptera species using ANN is possible assuming a sufficiently high number (tens) of specimens of each species is available for the ANN training. The methodology proposed is quite general and can be applied for all biological objects where it is possible to define adequate diagnostic characters and create the appropriate database.

The 1997 check list of Dolichopodidae of the Czech Republic and Slovakia has recently been reviewed and updated. The new species list includes 346 species with 22 species added as new to the fauna of the Czech Republic. While the check list itself is published elsewhere, largely unpublished new records of Hercostomus argentifrons, H. nigrilamellatus, Medetera adjaniae, M. melancholica and M. setiventris are presented here, together with data on their distribution in Europe and their biology and ecology. The status of the newly added Sympycnus desoutteri is discussed. H. argentifrons is recorded here for the first time from the Czech Republic (Bohemia; Moravia) and background information is given on its discovery. While the Czech fauna with 324 species can be considered well known, the fauna of Slovakia is definitely much richer than its current national list of 217 species suggests. In the latter country, in particular surveys of sandy habitats with heathland or peatmoor, saltmarshes, reedmarshes, humid forests on loamy soils, and of rothole and saprun microhabitats on trees might quickly yield new species records.

Five larval diets for laboratory rearing of Ceratitis capitata were tested. These diets were based on wheat bran, microcellulose, potato starch and agar. To evaluate the quality of diet, pupal rearing efficacy and pupal weight were checked. The best results were obtained with an agar based diet used for Manduca sexta laboratory rearing. To simplify the preparation and to reduce the cost of this diet, a new formulation was developed. Larvae reared on the new agar-based diet achieved higher pupal rearing efficacy than larvae reared on bran diet recently used in medfly mass rearing facilities. Heat treated medflies reared on the new agar-based diet achieved similar pupal rearing efficacy with heat treated medflies reared on bran diet. When testing population density, higher pupal rearing efficacy was again achieved on new diet. The highest pupal rearing efficacy was achieved with 100 eggs per 25 g of diet, lowest with 500 eggs per 25 g of diet. Concerning pupal weight, there was no difference in results achieved on Petri dishes with different larval population densities. Larvae reared on new agar-based diet reached better results than larvae reared on bran diet. The preparation of the new diet is simple and the cost is low, so it is good for laboratory tests and rearing.

Microhabitat selection and seasonal activity of the snake-eyed skink, Ablephaus kitaibelii fitzingeri, are compared to the two lacertid lizards (Lacerta viridis and Podarcis muralis) that co-occur in many of its habitats. The food composition of A. k. fitzingeri is also described. Significant differences in microhabitat selection and seasonal activity among the three species were found. The snake-eyed skink was associated with open grasslands, and with a low level of scrub, bare soil and rock cover. The microhabitat preference of L. viridis was quite similar to that of the skink, but with a higher preference for scrub. P. muralis occurred in places with greater rock and bare soil cover, and more scrub than A. k. fitzingeri. Activity of the snake-eyed skink decreased dramatically in summer, probably because of the reduced thermal inertia originating from the extremely small size of this species, but its seasonal activity overlapped with those of the lacertids. Stomach content analysis of the snake-eyed skink suggests that it is a generalist predator of small, mainly flightless arthropod prey. Competition with juvenile lacertids and predation by adult L. viridis are conceivable for the snake-eyed skink.

Predation on the non-native cyprinid, Pseudorasbora parva, was reported during an annual study in a small connecting canal about 40 km south of Brno, Czech Republic. Two cyprinid fish species (Pseudorasbora parva, Rutilus rutilus) were identified in the diet of 23 piscivores belonging to three native fishes (Perca fluviatilis, Sander lucioperca, Esox lucius). The relative abundance of prey fishes changed seasonally and was dominated by the topmouth gudgeon in spring. Thereafter roach incidence continuously increased. In contrast, topmouth gudgeon was found to be a more sized-available prey and was the most dominant prey species in the diet of all piscivorous fishes during the whole season. Strong piscivory towards this alien might be beneficial for its eradication or control to support ecological integrity in natural and modified seminatural habitats.

Apoptosis is a form of programmed cell death that plays an important role in small intestine ischemia-reperfusion (IR) injury. The aim of this study was to determine the total proportion of apoptotic cell death (apoptotic index) following injury induced by ischemia and during various subsequent reperfusion periods, total histopathological status and the intestine regeneration dynamics after the IR injury. Experimental animals, Wistar rats (n = 45) were divided into three experimental and one control groups. In the experimental groups 1 h ischemia was followed by 1, 4 and 24 h reperfusion. Intestinal ischemia was induced by superior mesenteric artery (SMA) occlusion. Segments of jejunum were stained with hematoxylin and eosin and studied immunohistochemically using M30 CytoDEATH and in situ TUNEL methods for apoptosis detection. Our experimental data showed that: (i) apoptosis is an important form of cell death in the small intestine after IR injury induced by SMA occlusion; (ii) maximum levels of histopathological damage and apoptotic index of mucosa occurred after 1 h ischemia and 1 h of reperfusion; and (iii) mucosa possesses great regeneration ability. The lowest levels of histopathological damage and apoptotic index were observed in the group with 1 h ischemia and 24 h reperfusion where, however, the highest mitotic index was present.

Spikes arriving at the synaptic connection produce short-term plastic changes of the synaptic efficacy. Model experiments have shown that paired-pulse facilitation attaining its maximum after a specific interval between a pair of arriving spikes might turn a “weak” plastic synapse attached to an integrate-and-fire neuron to a frequency-tuned device. Resulting computational capabilities create biologically plausible mechanisms of information processing relating to: (i) real-time identification of temporal patterns in a stream of random spiking activity (a recognition problem); and (ii) codetermination of the specific activity routing among neurons (an addressing problem) resulting in definite spatio-temporal patterns of the output activity (an input-output pattern problem).

Third part of chromosome number and DNA ploidy level reports from Central Europe comprising whole Carpatho-Pannonian region includes the data for following taxa: Scilla bifolia s. str. (2n = 18), S. bifolia agg. (2n = 36, 54), S. drunensis subsp. drunensis (2n = 36), S. drunensis subsp. buekkensis (2n = 36), S. kladnii (2n = 18) and S. vindobonensis (2n = 18) by J. Kochjarová from Austria, Slovakia and the Czech Republic (nos. 27–32); Campanula macrostachya (2n = ca 32) and Erysimum diffusum (2n = 14, 28) by E. Michalková from Hungary (nos. 33–34).