Volume 64, Issue 1

This review focuses on utilization of plant lectins as medical diagnostic reagents and tools. The lectin-related diagnostic is aimed at detection of several diseases connected to alteration of the glycosylation profiles of cells and at identification of microbial and viral agents in clinical microbiology. Certain lectins, proposed for or used as diagnostic tools could even recognize those cellular determinants, which are not detected by available antibodies. Broad information is presented on the lectinomics field, illustrating that lectin diagnostics might become practical alternative to antibody-based diagnostic products. In addition, the rising trend of lectin utilization in biomedical diagnostics might initiate a development of innovative methods based on better analytical technologies. Lectin microarray, a rapid and simple methodology, can be viewed as an example for such initiative. This technology could provide simple and efficient screening tools for analysis of glycosylation patterns in biological samples (cellular extracts, tissues and the whole cells), allowing thus personalized detection of changes associated with carbohydrate-related diseases.

DNA sequences can be used for the analysis of genetic variation and gene function. The high-throughput sequencing techniques that have been developed over the past three years can read as many as one billion bases per run, and are far less expensive than the traditional Sanger sequencing method. Therefore, the high-throughput sequencing has been applied extensively to genomic analyses, such as screening for mutations, construction of genomic methylation maps, and the study of DNA-protein interactions. Although they have only been available for a short period, high-throughput sequencing techniques are profoundly affecting many of the life sciences, and are opening out new potential avenues of research. With the highly-developed commercial high-throughput sequencing platforms, each laboratory has the opportunity to explore this research field. Therefore, in this paper, we have focused on commercially-popular high-throughput sequencing techniques and the ways in which they have been applied over the past three years.

The dynamics of MinD protein has been recognized as playing an important role in the accurate positioning of the septum during cell division. In this work, spot tracking technique (STT) was applied to track the motion and quantitatively characterize the dynamic behavior of green fluorescent protein-labeled MinD (GFP-MinD) in an Escherichia coli system. We investigated MinD dynamics on the level of particle ensemble or cluster focusing on the position and motion of the maximum in the spatial distribution of MinD proteins. The main results are twofold: (i) a demonstration of how STT could be an acceptable tool for MinD dynamics studies; and (ii) quantitative findings with parametric and non-parametric analyses. Specifically, experimental data monitored from the dividing E. coli cells (typically 4.98 ± 0.75 µm in length) has demonstrated a fast oscillation of the MinD protein between the two poles, with an average period of 54.6 ± 8.6 s. Observations of the oscillating trajectory and velocity show a trapping or localized behavior of MinD around the polar zone, with average localization velocity of 0.29 ± 0.06 µm/s; and flight switching was observed at the pole-to-pole leading edge, with an average switching velocity of 2.95 ± 0.31 µm/s. Subdiffusive motion of MinD proteins at the polar zone was found and investigated with the dynamic exponent, α of 0.34 ± 0.16. To compare with the Gaussian-based analysis, non-parametric statistical analysis and noise consideration were also performed.

Two species of the genus Choricystis (Choricystis minor and Choricystis chodatii) have been chromosomally surveyed. In addition, their ability to form symbiotic associations with the ciliate Paramecium bursaria was also investigated. Choricystis minor (clone #8931/1 from strain #8931) was isolated from a moss sample collected on Livingston Island, Maritime Antarctica. Choricystis chodatii (clone #3090/1) was derived from strain #3090 Chodat-type culture (loc. Switzerland, lake Geneva). The karyotypes of both species showed a haploid number of five chromosomes and shared some similarity in the chromosome lengths. The absolute chromosome lengths ranged from 1.25 µm to 2.95 µm. Studied species possess equal abilities for endosymbiotic associations with Paramecium bursaria. Descriptions of the species and a short discussion on their taxonomical status are given.

The current paper presents four lichen associations, including rare and endangered lichen species found in Lithuanian oak-woods. The structure of communities on the oak, characteristic of these localities, and geographical and sociological evaluation of communities are presented. Main reasons causing the extinction of rare species in the communities are mentioned as well.

Antimicrobial features of acetone, methanol and aqueous extracts of lichens of Cladonia furcata, Parmelia caperata, Parmelia pertusa, Hypogymnia physodes and Umbilicaria polyphylla were investigated by two different methods at the same time. Testing of antimicrobial activities of extracts from five species of lichens was performed by disc diffusion test in relation to Gram-positive and Gram-negative bacteria and fungal organisms, and through determination of minimal inhibitory concentration (MIC) by Broth Tube Dilution method. The obtained results indicated that acetone and methanol extracts of all investigated lichens in different concentrations manifested selective antibacterial and antifungal activity. That activity was more evident in relation to Gram-positive, than Gram-negative bacteria and fungal organisms. Acetone and methanol extracts of lichens Parmelia pertusa, Hypogymnia physodes and Umbilicaria polyphylla inhibited the growth of all tested microorganisms, most of all of lichens Cladonia furcata and Parmelia caperata. Although, the methanol extracts were generally the most active against the test organisms, the lowest MIC value was measured for acetone extract of species Cladonia furcata 0.39 mg/mL in relation to bacterium Bacillus subtilis. Aqueous extracts of investigated lichens were inactive against all tested organisms.

In the present study we report some preliminary results concerning the evaluation of antiproliferative activity on murine myeloma cells (P3X63-Ag8.653) of crude extracts of two common lichen species, Evernia prunastri and Xanthoria parietina. The results were evaluated by means of the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] test, which is commonly used to assess the activity of living cells through mitochondrial dehydrogenases. They indicated that extracts of E. prunastri had no effect, while those of X. parietina significantly affected murine myeloma cell proliferation, with a reduction down to 75% for methanolic extracts. This opens perspectives for deeper investigations extended also to other mammalian cell lines.

Growth and enzymatic activities of extraradical mycelia (ERM) of native mycorrhizal symbionts associated with three orchid species, Dactylorhiza fuchsii, D. majalis and Platanthera bifolia, were studied. ERM extracted from the mycorrhizosphere of these species showed features typical for fungi that form orchid mycorrhiza. In the first pot experiment, three different treatments were applied on tubers of D. fuchsii transplanted from a natural site: control (no specific treatment), reinoculated (surface-sterilized tubers reinoculated with mycorrhizal fungi-colonised roots), and benomyl (nonsterilized tubers treated with fungicide). However, no significant differences in ERM growth and intensity of root mycorrhizal colonisation at harvest were observed among these treatments. ERM associated with reinoculated D. fuchsii plants showed significantly higher alkaline phosphatase (ALP) enzymatic activity at week 36 than at week 24, but no differences were observed for NADH diaphorase activity. Benomyl application significantly reduced ALP activity in comparison with reinoculated plants at week 36. In the second experiment, plants of all three species were either untreated (control), or repeatedly treated with benomyl. Similarly to the results of the first experiment, benomyl application did not reduce the ERM growth of mycorrhizal symbionts associated with D. majalis and D. fuchsii. The low ERM growth associated with benomyl-treated P. bifolia was probably caused by poor root system development in this treatment. Significantly higher mycorrhizal colonisation was found for D. fuchsii compared to P. bifolia in control treatments at the end of cultivation. The ERM of native symbionts of the three orchid species studied seemed to have a different growth pattern over time and responded differently to fungicide application.

We studied the demography of Viola elatior, V. pumila, and V. stagnina, three rare and endangered Central European floodplain species, to (i) analyse variation in life-cycles among congeners and between regions (Dyje-Morava floodplains, Czech Republic; Upper Rhine, Germany), (ii) to define sensitive stages in the life-cycles, and (iii) to identify possible threats for population viability and species conservation. Matrix models were based on the fate of marked individuals from a total of 27 populations over two years. We analysed population growth rate (λ), stage distribution, net reproductive rate (R 0), generation time, age at first reproduction, and elasticity and calculated a life table response experiment (LTRE). Most populations were declining and λ did not differ between species or regions during the observed interval. Despite higher probabilities for survival and flowering in the Dyje populations, R 0 was higher in the Rhine populations. Also other demographic traits showed consistent differences between regions and/or species. Complex life-cycles and large variation in λ precluded unequivocal identification of sensitive stages or vital rates for conservation. Variation between regions may be a consequence of differences in habitat quality. Our results suggest that deterministic processes such as reduced management, succession, habitat destruction, and lack of disturbance through reduced or eliminated flooding present the strongest threat for the viability and persistence of populations of the three floodplain violets as compared with stochastic processes. However, the persistent seed bank of the species may buffer populations against environmental variation and represents a reservoir for recovery after resumption of suitable land-use management.

The relationships between the closely related Turkish endemic species, Viola sandrasea Melchior and V. kizildaghensis M. Dinç & Ş. Yıldırımlı, are discussed in the present study. Viola sandrasea has been known only from a few gatherings and insufficient description since 1939. Morphological and palynological properties of it are determined in detail and compared to its closest relative V. kizildaghensis in this study. The special habitats these species occupy, their typical associates and some other ecological features are also briefly discussed. The obtained data suggests that the morphological differences between them are sometimes subtle, but the petal colour, the leaf wrinkles and the pattern of indument are distinct. The pollen characteristics clearly appear to have taxonomic value. The pollen exine ornemantion of V. sandrasea characterized by the rugulate clearly differs from that of V. kizildaghensis characterized by psilate-perforate. Moreover, the two species are seasonally and geographically isolated from each other.

Five small water bodies located within the agricultural region of Wielkopolska (west Poland) underwent investigation. Periphyton samples were collected from various macrophyte habitats representing rush vegetation (in five water bodies), submerged aquatic plants (in three) and nymphaeids (in one): Palędzie — Ceratophyllum demersum, Potamogeton crispus, Typha latifolia; Batorowo — Phragmites australis; Piotrowo — Potamogeton natans, Ceratophyllum submersum, Typha latifolia; Tarnowo Podgórne — Typha latifolia; Dąbrówka — Zannichellia palustris, Potamogeton pectinatus, Phragmites australis. The main goal of the study was to determine the composition and abundance of the periphytic communities inhabiting various types of rush and water vegetation of five water bodies located within a mid-field landscape area. Diatoms such as Achnanthidium minutissimum, Amphora ovalis, Cocconeis placentula orNavicula cincta revealed significantly higher densities in the zone of elodeids, while green algae prevailed among nymphaeids. As a result of this study it was found that the epiphytic algae were characterised by much lower diversity in respect to a specific water body, though much greater diversity was observed in its relation to the type of substratum. Two types of habitats were distinguished — the first of simple build (helophytes and nympheids) and the second containing the complicated architecture of plant stems (elodeids).

In vitro culture is an important aid for ex situ conservation of rare, endemic or threatened plants. In this work, we establish an efficient method for the seed germination, seedling development, and axillary shoot propagation of Centaurea zeybekii Wagenitz. The seeds, collected from a wild population, were surface sterilised and cultured on various in vitro germination media. The effects of photoperiod and temperature on seed germination were also investigated. Germinations were obtained after 6 weeks in culture and the radicle emergence was evaluated as a main indicator. A high frequency of germination was obtained on distilled water supplemented with vitamines and 1 mg/L GA3. Although the seed germination frequencies were not affected by photoperiod, the highest germination frequency was obtained at 24 ± 2°C. A high frequency of axillary shoot proliferation was produced on MS medium supplemented with 1 mg/L BA. Then, the axillary shoots were separated and transferred to MS medium with or without plant growth regulators for rooting. Rhizogenezis was promoted after 6 weeks only in MS and 1/2 MS media containing 0.5 mg/L IBA. The rooting process was very slow and the percentage of shoot rooting was also very low (15%). The present study not only enables reinforcement of wild plant populations using ex situ growth of individuals, but it also helps to large number of aseptic seedling to use it in clonaly micropropagation studies.

Heat shock protein 90 (Hsp90), known as molecular chaperone, is involved in protein folding and assembly in the cell. In the present study, a full-length cDNA named Vitis pseduoreticulta heat shock protein 90 (VpHsp90) (GenBank accession Number:EU239815), encoding a heat shock protein 90, was obtained by degenerated primers and 3′-and 5′-RACE from Vitis pseudoreticulata according to our previously obtained EST sequence (GenBnak accession number:DV182112), putatively known as Hsp90. Comparison of VpHsp90 sequence has revealed that an open reading frame (ORF) consists of 2,100 bp nucleotides and the translated proteins of 699 amino acid residues. The molecular mass of VpHsp90 calculated from the deduced amino acid sequence was 80.2 kDa, Isolectric Point was 4.893, which is in close proximity of Hsp90. The maximum similarity of VpHsp90 at nucleotides level (85%) and protein level (96%) was found to be with Nicotiana tabacum. Phylogenetic tree analysis at both the nucleotides and amino acids levels indicates that Vitis pseduoreticulata, Nicotiana tabacum, and Arabidopsis thaliana Hsp90 sequences comprise one clade, which is closely related to Oryza sativa, Hordeum vulgare and Triticum aestivum Hsp90s. It may be reasonably concluded that VpHsp90 possesses the ancestral gene of Hsp90 similar to that of higher plant species.

The richness of endemic species is often recognized as an indication of the distinctiveness of certain local faunas and is used for the definition of conservation hotspots as well. Faunas of different animal taxa were considered in sets of contiguous geographical units. Comparing the faunas of different units in one set, we found an exponential increase in the number of endemics when plotted against the number of non-endemics. A model of independent stochastic population dynamics under the control of environmental oscillations produces random fluctuations in the ranges of species. Ranges of endemic species are supposedly narrower than ranges of co-occurring non-endemic species. In such a case, the flow of a random process leads to an exponential relationship between numbers of co-occurring endemic and non-endemic species. This process also produces an apparent positive correlation between total species number and the percentage of endemics.

A new soil mite species, Quadroppia (Coronoquadroppia) sezeki sp. n., is described. Adult mites were recorded from moss under a pine tree from a pine forest. The new species differs from the other species of the subgenus by bearing epimera III+IV with a wide key hole shaped inner border and the shape of frontal appendage of rostrum. The morphological features of the new species are also compared with those present in closely related species.

The epigeic spider communities of the Western Carpathians are characterised by a high number of species. The great influence of Pannonian elements on the coenoses of the Carpathian hills is evident. The habitats of the Western Carpathians differ from each other, and several types of microclimatic conditions are there. These facts affect the spider fauna. The epigeic spider communities of the western part of the Carpathians, represented by the Devínska Kobyla hill, were characterized by the constant presence of four species at all study sites — Dysdera erythrina, Agroeca cuprea, Pardosa lugubris and Trochosa terricola. Principal components analysis confirmed the high proportion of Trochosa terricola, Eresus moravicus and Scotina celans in the epigeic spider community of the steppe. In contrast, the presence of Coelotes inermis, Histopona torpida and Haplodrassus silvestris was important in the oak forest. Bray-Curtis analyses confirmed that the spider community of the oak forest differed. The migration and penetration of the species with high vagility was an important fact that affected the spider community in the ecotones. Based on the analysis of the araneocoenoses in the Western Carpathians, spiders were grouped into four categories — leaders, typical spiders, specific spiders and tourist species.

Morphological colour variation in Idotea baltica basteri Audouin, 1827 (Isopoda: Crustacea) is reported for the first time on populations from a brackish water lake (Kucukcekmece Lagoon) in Turkey. According to pigmentation characteristics, the isopods were described and sorted to ten different colour morphs which are named as maculata, maculata-lineata, uniformis, uniformis-lineata, immaculatum, immaculatum-lineata, nigrum, nigrum-lineata, albafusca, and albafusca-fasciatum. The similarity of the morphs was compared by defining digit codes to some of the visual pigmentation characteristics and calculating the Bray-Curtis similarity index of the colour morphs. The albafusca and albafusca-fasciatum forms were found to be the most similar to each other (similarity 96%), whereas immaculatum forms were considered the most different from maculata-lineata (similarity 50%). The morphs named maculata and maculata-lineata, having the highest abundance with 70%, belong to the most successful forms which may achieve crypsis by background matching in their living habitat in Kucukcekmece Lagoon which has sandy and rocky bottom surface area.

Samples of dying branches from 121 trees of Quercus pubescens, Q. cerris, and Q. petraea were taken from 102 sites in Seslerio autumnalis-Quercetum petraeae and Ostryo-Quercetum pubescentis forests. After nine months’ rearing in eclectors, branches were cut to 20-cm sections, and signs of infestation as well as larvae and adult insects were noted. We analysed 395 sections of branches from Q. pubescens, 125 sections from Q. cerris, and 85 sections from Q. petraea. We found 44 adult insects: 45% were from the Cerambycidae family (Callimus angulatus ssp. angulatus, Poecilium alni, and Exocentrus adspersus), 18% were from the Scolytinae subfamily (Scolytus intricatus, Xyleborus dispar), 4% from the Buprestidae family (Coraebus florentinus), 4.5% from the Cleridae family (Tilloidea unifasciata), and 28.5% from other families. In addition, 145 larvae were found: 50% from the Cerambycidae family, 39% from the Buprestidae family, 4% from the Scolytinae subfamily, and 7% from other families. Significant difference in the abundance of adult insects and larvae with regard to the diameter of branch sections and the host species were found. 78% of dying branches showed signs of infestation. Species of the Buprestidae, Scolytinae, and Cerambycidae represent important factors in the decline in oak vitality in the lower Karst region of Slovenia.

The cooperation and aggression between five laboratory colonies of Monomorium pharaonis were compared using an aggressiveness test and pupa-carrying test in laboratory arenas. The colonies were derived from field collections in different parts of Europe and USA. Generally, inter-colony aggressiveness was low and acceptance of pupae from other colonies was high. Workers from one colony (Lužiny, CZ), however, frequently displayed aggressive behavior when paired with workers from other colonies, and the Lužiny pupae were avoided by workers of other colonies in pupa-carrying tests. Behavioral tests were only partly consistent with the phylogenetic relatedness of ants because the Wisconsin colony (USA) grouped with the Lužiny colony (and not with the other three colonies) in the phylogenetic analysis but grouped with the other three colonies in the behavioral tests.

The descriptions of pupae of three Western Palaearctic species of Erebidae (subfamily Herminiinae), namely Idia calvaria (Denis et Schiffermüller, 1775), Pechipogo flavicrinalis (Andreas, 1910), and Nodaria nodosalis (Herrich-Schäffer, 1851), and three species of Noctuidae (subfamilies Acronictinae and Bryophilinae), namely Oxicesta geographica (F., 1787), Simyra dentinosa (Freyer, 1839), and Cryphia ochsi Boursin, 1940 are given. The main morphological features are described, compared with those of the related taxa and also illustrated in 57 line drawings. The systematic status and position of the described species present in the current systems are also discussed from the point of view of morphology of their pupae.

Atatürk Dam Lake is an important ecosystem located in south-eastern Anatolia in Turkey. In this study, concentration of Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn were determined in kidney, liver, gill and muscle of Tor grypus which has great economic value in Atatürk Dam Lake. Heavy metal levels in fish samples were analyzed by inductively coupled plasma spectroscopy (ICP). The metal accumulation in the liver, kidney and gill of Tor grypus was found to be quite high in comparison to the muscle. All metal levels detected in tissues were the limits for fish proposed by FAO/ WHO, EU and Turkish Food Codes and safe within the limits for human consumption in the edible parts of fish species in the region.

From 2003–2006, research on the breeding distribution of the white-tailed eagle (Haliaeetus albicilla) was conducted in Croatia in order to assess the size of the national population. In 125 locations, clear signs of breeding activity were found. An additional 10 presumably active territories were detected but it was not possible to locate the exact position of the nests and confirm the breeding. Based on this, it is concluded that the national breeding population is not less than 135 breeding pairs. The present distribution can be compared with previous reports with the exception of the area along the Ilova and Lonja rivers that have never been reported as an important breeding site. Analysis of the characteristics of 138 nest positions as well as preferences/avoidance of specific structural features were performed. The results showed that white-tailed eagles prefer to build their nests on pedunculate oaks, narrow-leafed ash and white poplars with the greatest preference for mature trees with a diameter above 92.5 cm. The minimal distance between two active pairs was 348 meters. More than 50% of the national population breed less than two km from a large water area and 95% of the population less than four km. More than 95% of the population breed at altitudes lower than 140 m above sea level and are further than one km away from the nearest human settlement, regardless of the availability of forests. According to several parameters (distance to a large water area, elevation, forest presence, distance to the nearest settlement, distance to highways and railways) geographic information system (GIS) helped to determine potential white-tailed eagle breeding areas.

The structure of a breeding bird assemblage of a primeval alder swamp in the Šúr National Nature Reserve (the Danube Basin) was studied in the period 1992–1995. A 16 ha forest interior study plot was established for bird censusing. Population abundances were estimated by a combined version of the mapping method from the end of March to the beginning of July. Altogether, 42 breeders were recorded and the mean total density of species total across years was 125.15 ± 12.73 pairs/10 ha (CV = 10.17%). Two species were eudominant (≥ 10%): Sturnus vulgaris and Anas platyrhynchos, and six species were dominant (5% ≤ 10% <): Ficedula albicollis, Fringilla coelebs, Phylloscopus collybita, Parus major, Sylvia atricapilla, Erithacus rubecula. The Shannon diversity index (H′) varied between 3.98–4.10 bites. The evenness index (J′) reached values between 0.79–0.81. Expected species diversity in a random sample of 100 pairs calculated by rarefaction [E (S 100 pairs)] was 21.35 ± 0.92 species derived as a mean value from the years 1992–1995. The mean rarefaction estimate on the area [E (S 10 ha)] was 22.75 ± 1.58 species. Bird species richness and diversity were significantly higher in the study plot in comparison to the mean value of European wet alder forests. These results are comparable with the values of structural assemblage parameters of the primeval stands dominated by alder within the Białowieża National Park, Poland.

The in situ identification of carbohydrate structures in Trichinella spiralis intestinal larvae, adults and L1 muscular larvae was carried out by lectin histochemistry, with emphasis on the O-linked glycans. The absence of reactivity with two lectins-TML and MAL indicated that Trichinella spiralis does not synthesize sialic acid. Reactivity with HPA, VVL-B4, PNA and UEA-I staining suggested that T. spiralis synthesizes and expresses on its cuticle O-linked glycans analogous to Tn-antigen (GalNAc-α-Ser/Thr), T-antigen (Gal-β1,3-GalNAc-α-Ser/Thr) and also structures analogous to A-blood group antigens (GalNAc-α1,3-Gal-β1,3(4)-(Fuc-α1,2-)-R). Expression of the saccharidic moieties is stage-specific. Blood group-A and T-antigen structures were identified on the cuticle of the intestinal and muscular larvae. The Tn-antigen structure was missing in the intestinal larvae. Appropriate ligands for WGA were not identified in the adult individuals. The obtained results may contribute to a better understanding of the glycobiology of this parasitic nematode in relation to occupation of its intracellular niche. The presence of saccharidic structures analogous to some of those expressed on the intestinal epithelial cells may serve as a protective shield on the surface of the parasite.

N-acetyl-5-methoxytryptamine or melatonin is a multifunctional molecule. The main physiological function, at least in vertebrates, is to transduce to the animal the photoperiodic information and regulate rhythmic parameters. But studies have also observed the action of this molecule on pigment migration in ectothermic vertebrates. Thus the aim of this paper was to investigate in vivo and in vitro the influence of melatonin on the pigment migration in melanophores of the crab Neohelice granulate. Injections of melatonin (2 × 10−9 moles · crab−1) at 07:00 h or 19:00 h did not affect (p > 0.05) the circadian pigment migration of the melanophores in constant darkness. Additionally no significant pigment migration (p > 0.05) was verified in normal and eyestalkless crabs injected with melatonin (10−10–10−7 moles · crab−1) during the day or night. In the in vitro assay, the response of melanophores to the pigment-dispersing hormone in eyestalkless crabs injected with melatonin (2 × 10−9 moles · crab−1) 1 and 12 hours before the observations did not differ (p > 0.05) from the control group (injected with physiological solution). These results suggest that melatonin does not act as a signaling factor for pigment dispersion or aggregation in the melanophores of N. Granulate.

This study investigates the differential expression of lactate dehydrogenase (LDH) isoenzymes in the genus Channa using PAGE. With the help of obligate air-breathing, all of the selected species can sustain water deprivation to varying degrees. In subunit composition and higher electrophoretic mobility of LDH-A4, the profiles of channid species were similar to other teleosts documented in the literature. However, inter- and intra-species differences, with particular reference to aerobic/anaerobic metabolic options, existed. Whereas glycolysis in Channa punctata appears to depend largely on aerobic LDH-B and partly on anaerobic LDH-A, metabolism in C. gachua, C. striata and C. marulius depends exclusively on the activity of anaerobic LDH-A. Expression of the third locus Ldh-C was recorded in the eyes of C. marulius, in addition to C. gachua. Heat inactivation experiments reveal species differences between LDH isoenzymes and a general order of the relative stabilities: LDH-C > DH-B > LDH-A. Metabolic and evolutionary implications of the findings have also been discussed.

The effect of four sublethal concentrations (from 0.01165 to 0.3025 µg per insect, responding to LC10/10, LC10, LC30 and LC50) of the organophosphorus insecticide, fenitrothion, on Spodoptera exigua (Lepidoptera, Noctuidae) larval development and ultrastructure of follicular cells of adult female moths was tested. The most prominent malformations were: breaks in the cuticle, increased mortality during larval/pupal molting, alterations of nuclear envelope, changed heterochromatin pattern and abnormalities in mitochondrial ultrastructure. Some of the alterations are dose-dependent, but the other ones, like organization of rough endoplasmic reticulum (RER), show non-linear response: cells exposed to lower concentrations (LC10/10 and LC10) of the insecticide possessed clusters of accumulated RER, which were absent within the cells exposed to higher ones (LC30 and LC50). Formation of the eggshell was also markedly postponed within the groups exposed to lower doses. These findings prove that even minute amounts of xenobiotics may cause significant changes within exposed populations.

The fact that leukaemic cells are primarily or secondarily resistant to cytostatics is a serious phenomenon, which leads to the failure of chemotherapy of malignant diseases in clinical practise. Some detoxification and transporting systems are responsible for the generation of chemoresistance on the cellular level and the decrease of effectiveness in treatment. In vitro testing of chemoresistance of leukaemic cells is presently an inseparable component of “tailoring” therapy in the developing field of predictive oncology. The aim of this work was to estimate profiles of drug resistance, based on the predictive in vitro test, and to help in choosing the most effective cytostatic. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoline (MTT) assay was used, based on the direct effect of cytostatics on the viability of leukaemic cells in vitro. The number of living leukaemic cells was evaluated by a computer program, where LC50 (concentration of cytostatics lethal to 50% of leukaemic cells) was established from the achieved dose-relation curves. Seventy-one samples of leukaemic cells isolated from the patients’ peripheral blood or bone marrow were examined. All samples were tested to 3 cytostatics minimally. It was found by the in vitro assay, that resistance to dexamethasone, prednisolone, etoposide and vincristine is increased in patients with acute myeloid leukaemia disease, compared to the acute lymphoblastic leukaemia patients. In patients with a relapsed disease population, leukaemic cells are highly heterogeneous in the MTT assay. It was concluded that the MTT assay can be used to study drug interactions in vitro in leukaemia samples. The type of interaction was highly different between patients, and depended on drug concentrations.

The osteoprotegerin (OPG)/receptor activator of nuclear factor-κB ligand (RANKL)/receptor activator of NF-κB (RANK) system plays an important role in the pathogenesis of metabolic bone diseases. This study is aimed to investigate effects and mechanisms of RANKL gene silencing on the function of human osteoblast-like MG63 cells by RNA interference using a lentivirus-based small hairpin RNA (vshRNA) delivery system. After RANKL-specific vshRNAs were designed, constructed and transfected into MG63 cells, changes in the expression levels of RANKL mRNA and protein were determined by Western blot and RT-PCR, respectively; changes in cell activity and cell cycle distribution were examined by thiazolyl blue tetrazolium bromide assay and flow cytometry. The expression levels of RANKL mRNA and protein in MG63 cells were reduced by transferring RANKL-specific vshRNAs. Compared to cells infected with negative control virus, the proliferation of cells infected with the recombinant virus was more likely to be inhibited. Furthermore, the cell cycle of MG63 was altered, with the number of G1 phase cells decreasing significantly (P < 0.05). RANKL-specific vshRNAs can significantly inhibit the expression of the target gene in MG63 cells. RANKL gene silencing can inhibit the proliferation and alter the cell cycle of MG63 cells. Our findings suggest that RANKL might play an important role in the regulation of growth and cell cycle of MG63 cells.