Volume 72, Issue 11

Paleolimnological research in the territory of Slovakia has relatively short history with the beginning in the 1990s primarily as a response to the ecological changes in mountain lakes due to air pollution and missing data from the pre-industrial period as well as a need to quantify anthropic impact on the landscape. In the present paper, we reviewed 53 publications, including research papers, monographs, conference abstracts and theses, dealing with aquatic proxies in Holocene lake sediments to identify (1) the location of surveyed sites, (2) proxies used, and (3) the object of the paleolimnological reconstructions. The vast majority (37) of the analysed paleolimnological localities (51) took place in the High Tatra Mountains, focusing on the alpine lakes of glacial origin. The most frequently used biological proxy was Chironomidae remains (44%). In contrast, Cladocera, Ostracoda, loss-on-ignition, molluscs and stable isotopes were used only in 10–12% of studies. Most studies (84%) focused on a simple ecological reconstruction of the past environment, and only 24% of the studies attempted to reconstruct specific ecological variables, properties and processes, such as acidification (10%), temperature (8%) and eutrophication (4%). Only 2% of the studies used molecular techniques. Sixteen out of the 51 cores analysed (i.e., 31%) were dated by radiometric methods. In the end of our review we pinpoint the weaknesses of the local paleolimnological research and sketch possible future directions.

Proteoglycan-like (PG) domain of carbonic anhydrase IX (CA IX) is a small-sized acidic domain (8.3 kDa; pI ∼ 3.5) with a very unusual amino acid composition. More than one third of its sequence consists of acidic amino acids and it contains no aromatic residues. It has been revealed that it holds one of the key roles in oncogenetic mechanisms, in which CA IX participates. However, it has not been structurally characterized yet. With these prospects, we developed an expression system of recombinant PG domain production in the form of a fusion protein using thioredoxine (Trx) as the fusion partner in Escherichia coli . We performed three steps during purification process utilizing affinity chromatography, anion-exchange chromatography and gel filtration; and the highest purity we reached was 93.7% with protein concentration 23.3 mg/L. Hypothetical yield of the recombinant protein would be 0.581 mg/L of culture. The applicability of the fusion protein Trx-PG for further structural studies was confirmed by in silico structure prediction tool and ELISA.

We analysed a dataset composed by the frequency of 79 epiphytic lichens recorded on 243 oaks distributed in 29 oak dominated stands of the Western Carpathians, representing areas with high environmental quality up to disturbed environments. Lichen diversity indices (based on total frequencies and on functional and morphological groups) were used as indicators of the response to air quality and forest management. The level of air quality in these categories was a function of NO x , SO 2 and particulate matter. From management viewpoint the units were classified as managed (areas subjected to periodic logging) and semi-natural (which currently have the status of National Nature Reserve and where eventual management practices date back to the history). The results showed that higher environmental levels of pollution were associated to the decrease of fruticose and in general pollution sensitive species, while low environmental levels of pollution were associated to the diffusion of common nitrophilous lichens as well as to a higher share of fruticose and in general pollution sensitive species. As far as forest management is concerned, a higher share of fruticose and sensitive species was associated to semi-natural and natural stands, whereas a higher share of foliose (pioneer and tolerant) species was associated to the managed stands.

Clonostachys rosea is an important biological control agent against various plant pathogenic fungi. We collected – necrotic ponderosa and Coulter pine needles and obtained C. rosea – like fungal colonies. Cultures were identified as C. rosea and their morphological characteristics were described.

The lack of natural references of dry-mesic oak forests creates conceptual obstacles for their conservation and close-to-nature management in Central Europe. Single-tree inventory was used to investigate stand characteristics and old-growth attributes of a Quercus petraea (Matt.) Liebl. and Q. cerris L. dominated, 120-year-old stand in a Hungarian forest reserve, abandoned approximately 40 years ago. All individuals were recorded with diameter ≥ 5 cm on the 3 ha study site. Logs, stumps, saplings and seedlings were also surveyed. 582 woody stem/ha belonging to 14 species were measured with single-tree inventory. Basal area values showed the total dominance of oaks in the canopy layer (99%) as a legacy of the long-term human exploitation. In contrast, young individuals of oak species were almost missing, and Acer campestre L. dominated the lower layers, indicating the transitional nature of the stand. Diameter classes showed a marked bimodal distribution. Both the abandonment of the reserve and the precedent severe oak decline contributed to the relatively high amount of dead wood (46 m 3 /ha). Four decades of abandonment is rather short time interval to generate a diverse forest composition and structure in a mature dry-mesic oak forest. The dense regeneration and shrub layer and the upsurgence of admixing species indicate the shift towards the uneven-aged and compositionally more diverse old-growth oak forest state. Among the structural forest features the dead wood had similar values as old-growth forests. If the recent trend continues, the studied oak forest develops towards a mixed forest with significantly lower ratio of oak species.

Invasive plant species are a growing concern in global biodiversity management. In this study, we tried to determine whether invasive plants could spread from railway track into the surrounding areas and focused on the factors responsible for the invasive plant species composition, richness, and abundance. We sampled invasive species along transects located outside a railway route and recorded the invasive species richness and abundance at a distance of < 5 (control), 5–10, 10–15, and > 15 m away from the railway edge in three geographical regions in SE Poland. We revealed that the mechanism allowing alien species to invade was complex. Canonical correspondence analysis (CCA) explained 30.9% of variance. On a regional scale, the invasive flora composition in the surroundings of railway habitats was determined by abiotic environment diversity (mean annual air temperature, mean annual precipitation), but not habitat-related factors (soil type, soil properties). On a spatial scale, the traffic volume, the distance from the railway area, the distance from build-up areas, and the interaction of these factors affect the invasive species composition away from railway areas. The invasive species spread from railways to the surrounding vegetation, therefore, regular monitoring of railway areas should be a framework for assessing the risk of invasive species.

The main goal of this study was to evaluate the role of polysaccharide and phospholipids on the surface activity of model root mucilage in its liquid and solid phases. Polygalacturonic acid (PGA) and Phosphatidylcholine (PC) were used as representative polysaccharide and phospholipid, respectively. The surfactant performance of the PC found to be superior relative to the PGA – reducing the surface tension (ST) down to 32.9 mN m -1 at 0.04 mg mL –1 while the PGA reduced the ST to 47.7 mN m -1 at 15.9 mg mL –1 . The minimum ST of the PC/PGA mixture was similar to the pure PC for total concentration of 2 and 10 mg mL –1 at PC mass-fraction of 0.01 and 0.1, respectively. The PC, PGA and the PC/PGA mixtures induced subcritical soil water repellency to sand particles. The maximum contact angle (CA) obtained for the PC and PGA were 85° (at 0.7 mg g -1 ) and 75° (4.5 mg g -1 ), respectively. The maximum CA obtained for the PC/PGA mixture was 85°, depending on the total concentration and the PC mass-fraction. It is suggested that during the rehydration of sand coated with air-dried mucilage, water flow at the rhizosphere may initially affected by the induced water repellency. However, with elapsed time, dissolution or detachment of surface-active constituents of the mucilage from the sand surface into the soil solution may reduce the ST of the soil solution at the liquid-air interface and consequently the CA at the wetting front.

Zizania latifolia is a perennial aquatic vegetable of the Poaceae family. Chilling temperatures significantly affect stem enlargement and harvest time of Z. latifolia . Cold tolerance improves growth and harvest time at low temperatures. The study aimed to elucidate the proteomics and physiological differences between cold stress-tolerant (Longjiao 2) and -sensitive (911) Z. latifolia varieties. Superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) enzyme activities, as well as levels of malondialdehyde (MDA) and soluble sugars were determined. 2D gel electrophoresis and MOLDI-TOF MS were used to detect the differentially expressed proteins at normal and lower temperatures. Significant differences were observed for relative SOD, CAT, and POD enzyme activities, as well as for soluble sugar levels when comparing the two varieties grown at different temperatures. The results indicated that different responses of the two variety to chilling stress involved previously described physiological mechanisms. Proteomic changes under chilling stress suggested that key pathways including photosynthesis, energy metabolism, defense mechanisms, and RNA processing were differentially regulated in the chilling tolerant variety. Our findings provide a basis for the generation of new varieties Z. latifolia that could be capable of optimal growth under chilling stress, even in the context of global climate change.

Lettuce ( Lactuca sativa L.), a leafy vegetable belonging to Asteraceae family, possesses sedative-hypnotic, anticonvulsant, anti-inflammatory, antinociceptive, anticancer, and antioxidant properties. Currently, only a few microsatellite markers have been developed from lettuce, which hindered the genetic research. In the present study, EST-SSRs were developed and characterized from 81,518 expressed sequence tags of lettuce. The results showed that tri-nucleotide motif (40.789%) was the most abundant, followed by di-nucleotide (33.109%) and hexa-nucleotide repeats (16.295%). Among 100 primer pairs designed according to SSR-containing sequences, a set of 35 could give out reproducible and clear amplification products, and 15 were polymorphic. These novel microsatellite markers were used to evaluate genetic diversity of lettuce and its related species. The number of alleles per locus ranged from 3 to 11, with a mean value of 6.2. The expected heterozygosity ( H E ) and observed heterozygosity ( H O ) ranged from 0.5141 to 0.8346 and 0.0484 to 0.6000, respectively. Eleven could be transferable to three related species, including Artemisia argyi H. Lév. & Vaniot, Erigeron annuus (L.) Pers., and Dendranthema morifolium (Ramat.) Tzvel. These novel microsatellite markers will not only enrich current resource of genetic markers, but also be useful in the following cultivar identification, germplasm evaluation, and marker-assisted selection of L. sativa and related species.

Heat shock protein 40 (HSP 40), as an essential co-chaperone of HSP70, has been poorly investigated in planarians. In this study, a full-length cDNA of a HSP40 gene from planarian Dugesia japonica (designated Djhsp40 ) was identified and its expression pattern during planarian regeneration was examined. Its cDNA is 1,378 bp, which includes an open reading frame of 1,236 bp encoding 411 amino acids with a molecular weight of 46.2 kDa. DjHSP40 contains three conserved DNAJ domains belonging to the Type I HSP40/DNAJ family. After amputation is performed, Djhsp40 is up-regulated and the transcripts are specifically expressed in the blastema, and its inhibition by siRNA strongly delays the blastema growth. Interestingly, the silencing of Djhsp40 does not affect the expression level of neoblast-specific proliferation marker ( Dj-mcm2 and DjpiwiB ), but reduces Dj-mt-mmpA expression significantly. Our work provides a crucial clue for the study of the function of HSP40 in planarian regeneration.

A new species of Bothropolys Wood, 1862, Bothropolys jiuensis sp. n. (Lithobiidae: Ethopolyinae), and a new record of the species Lithobius magnitergiferous Zhang, 1996 (Lithobiidae: Lithobiinae) are described. The new species originates from JiuZhaigou County of the Qinghai-Tibet Plateau, and is distinguished from two recorded congeneric species by having 13–31 ocelli; porodonts situated between the outer two teeth; posterior triangular projections on TT9, 11, 13 distinctly; and no accessory claws in legs 14–15. A key to the Chinese species of Bothropolys is presented. Lithobius magnitergitiferous was collected in QuMalai County, Qinghai Province and shows TT7, 8 extended distinctly and larger than the other tergites. A key to the Chinese species of Lithobius Leach, 1814 is presented.

It is well established that hedgerows provide habitats, which cannot be found in otherwise coherent agricultural landscape. However, different organisms vary in their response to various environmental variables of hedgerows. The aim of this paper is to study how are the myriapod species richness and community composition related to biological and morphological parameters of farmland hedgerows. We expected diversity to be related to size of hedgerows and community composition to be primarily affected by their biological attributes. We studied myriapod communities in 20 hedgerows located in close vicinity of Žibritov village (the Carpathians, Slovakia) in Central European upland farmland. Characteristics of the hedgerows were either measured in the field or derived from GIS layers. We used univariate and multivariate correlation methods to investigate the effect of hedgerow characteristics on diversity and composition of myriapod communities. Surprisingly high taxa numbers of myriapods were found within small area. Richness of millipedes showed negative relationship with hedgerow plant community diversity, hedgerow length and soil depth and weak positive relationships with a degree of shrub layer defoliation. Centipede richness was mainly related to diversity of hedgerow tree layer. We did not find any significant relationship between centipede communities’ composition and environment. Length of hedgerows was the only variable showing significant relationship with millipede communities’ composition. Despite small area, hedgerows represent important habitats for several species of centipedes and millipedes and show specific composition of their taxocoenoses determined by hedgerows attributes. Farmland hedgerows appear to be important habitats for soil organisms, which may be further beneficial for surrounding landscape.

Only a few families have been recognized in the 320 million year history of cockroaches. The 31 st one, represented by monotypic Ol xiai sp. n. from the Myanmar amber is described here. It inhabited lowland forest near the shore. It possesses several unique morphological features including bipectinate antennae, round terminal palpomeres, sophisticated forewing shape with broken R and CuP forming a distinct unsclerotised area and with RS branched to both sides, and a huge foretibial spur. The new genus also displays several putatively apomorphic reductions (short antennae, rudimental femoral terminal spine, short pronotal chaetae, minor arolia, short palps). Synapomorphies with early ant-nest parasites (short antennae, dense pronotal setosity, irregular spacing of wing veins, indication of maculated colouration) suggest an origin from Blattidae or ancestral Mesoblattinidae during the Early Cretaceous. No plesiomorphies are apparent. An attached mite documents a phoresy on males. A syninclusion of a coccoid male is not ecologically informative at the given state of knowledge.

Freshly hatched larvae represent critical but little studied phase in insect development. In butterflies inhabiting seasonally harsh environments, such as mountains of temperate regions, the larvae have only short time available for feeding before entering diapause. In the species rich genus Erebia , post-diapause larvae activate and feed exclusively in evening and late night hours, whereas the diurnal schedule and modifying effects of weather of pre-diapause larvae are practically unknown. Using captive observation with a transplant between alpine (2000 m) and mountain (800 m) altitudes, we studied larval activity of two species, alpine Erebia cassioides and lower-altitude E. aethiops , in Austrian Alps. Both species activated and fed both during day time and after the sunset, contrasting with the situation in post-diapause larvae. Both activity and feeding were strongly and species-specifically affected by temperature, cloudiness and humidity. Additionally, the loweraltitude E. aethiops , but not the alpine E. cassioides , restricted activity and feeding in the alpine altitude, indicating that some factors particular for alpine environments, possibly increased UV load or reduced air pressure, may limit the ability of this mountain species to develop above its vertical distribution limit.

Carniolan honeybee ( Apis mellifera carnica ) is considered an endangered subspecies of Apis mellifera in Slovakia due to its illegal crossbreeding with allochthonous subspecies. Despite more than a century-long tradition of Slovak beekeeping, genetic research of Slovak honeybees has not yet been done. The aim of our study was to develop a molecular approach to characterize Carniolan honeybee population using polymorphic microsatellite markers. A total of 79 samples of Slovak Carniolan honeybee from 19 regions of Slovakia were analyzed together with 85 reference samples of Apis mellifera mellifera, Apis mellifera macedonica, Apis mellifera ligustica and Apis mellifera hybrid (Buckfast). Ten highly informative (PIC > 0.5) microsatellite markers running in two multiplex PCR reactions were used for molecular analysis of bees. The mean number of alleles per locus ranged from 4.6 ( A. m. mellifera ) to 11.5 ( A. m. carnica ). Bayesian and frequency-based methods implemented in software ‘GeneClass’ (v 2.0.h) have been used to assign individuals to populations. We were able to successfully assign 95% of individuals in actual sample size. After simulating 100,000 individuals using Bayesian Markov chain Monte-Carlo resampling, we correctly assigned 88% individuals to five tested populations. Population genetic structure was inferred through the Bayesian clustering method incorporated in software ‘Structure’ (v 2.3.4). Overall, 92% of the individuals were assigned to the same subspecies as originally declared. According to Structure analysis, Slovak population of A. m. carnica appears to be separated into two clusters, showing the relatively high level of gene flow from A. mellifera hybrid (Buckfast) and A. m. macedonica subspecies. It might be caused by rebuilding the Slovak population after colony losses using imported subspecies of the honeybee.

The alarming pace of loss of global biodiversity gets major attention worldwide, and amphibians are by far the most endangered group of vertebrates. IUCN estimates that 41% of amphibian species are threatened. The year 2008 was named as “The Year of the Frog” to raise both awareness and funding of mainly ex situ programs needed to mitigate the amphibian extinction crisis. Zoos and aquariums are expected to participate in ex situ conservation breeding programs of endangered species. Number of publications documents the increasing concern in amphibian captive breeding programs. In this paper, we reviewed the current effort of public zoological gardens in Czech Republic on amphibian keeping and breeding. Our results show that species collections reflect combination of the availability of species in the pet trade and the availability of surplus animals from other zoos, but not the needs of amphibian conservation.

This study describes the craniometrical population pattern of the European golden jackal ( Canis aureus moreoticus ) populations inhabiting the southeastern part of its European range. The craniometric analysis was carried out using 18 linear skull and dental parameters. The analysis included 167 adult individuals (78 males and 89 females), originating from five populations inhabiting Bulgaria, Serbia and Hungary. The expected high degree of craniometric sexual dimorphism was confirmed. The multivariate differentiation analyses of linear characters were carried out separately for males and females from the five analyzed populations. The results are suggesting that Multiple Group Discriminant Analysis is able to “build” a model which classifies both males and females to the population from which they originated. The general pattern of craniometrical similarity in both males and females golden jackals in investigated area is not strongly bounded to the geographic origin of their population. Comparative population analysis of linear skull and dental parameters of golden jackals in southeastern Europe provided important information for future population differentiation studies across the whole golden jackal range.

Fucoidan is a sulfated polysaccharide widely distributed in brown seaweed. It exhibits several bioactivities, such as anti-cancer, anti-tumor, anti-microbial, anti-diabetic and anti-oxidant properties. However, the effects of fucoidan in chondrocytes are not well established. Previously, we have reported in vitro and in vivo anti-inflammatory effects of fucoidan. In this study, we evaluated the effects and regulatory mechanism of fucoidan derived from Undaria pinnatifida on the cyclooxygenase-2 (COX-2) and type II collagen in rabbit articular chondrocytes. Using western blotting and alcian blue staining, respectively, fucoidan was shown to induce type II collagen and sulfated proteoglycan in a dose- and time-dependent manner. Moreover, fucoidan inhibited the COX-2 expression in a dose- and time-dependent manner and increased the phosphorylation of extracellular signal-regulated kinase (ERK), p38, and AKT kinases in chondrocytes. The inhibition of p38 and AKT using SB203580 and LY294002, respectively, in the presence of fucoidan decreased the expression of type II collagen. However, ERK inhibition using PD98050 stimulated type II collagen expression. Fucoidan increased COX-2 expression in the presence of inhibitors of ERK, p38, and AKT kinases. These results conclusively suggested that fucoidan regulated type II collagen expression via the p38 and AKT pathways, and COX-2 expression via the p38, ERK and AKT pathways in rabbit articular chondrocytes. Moreover, given its ability to mediate cell differentiation and exert anti-inflammatory activity, fucoidan may represent a potential therapeutic substance for use in inflammatory conditions, including arthritis.

Esophageal sensory and motor nerves contribute to the symptoms of eosinophilic esophagitis, however, the mechanisms of this neural dysfunction are essentially unknown. We addressed the hypothesis that eosinophilic inflammation in the esophagus alters production of the key regulators of neural function neurotrophins and neurotrophic factors. We developed and optimized the model of allergic eosinophilic inflammation of esophageal mucosa induced by localized administration of allergen ovalbumin into the esophagus in ovalbumin-sensitized guinea pigs. We evaluated changes in expression of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), glial cell-derived neurotrophic factor (GDNF) and artemin in esophageal mucosa by quantitative RT-PCR. We found that the administration of ovalbumin into the esophageal wall of sensitized animals induced a massive eosinophilic infiltration restricted to esophageal mucosa (3 ± 1 vs. 97 ± 23 eosinophils per high power filed). This inflammatory response altered the expression profile of selected neurotrophic factors. The BDNF mRNA was increased (to 200%), artemin mRNA was decreased (to 50%) while NGF and GDNF was not changed. We conclude that a strong eosinophilic inflammation can be induced in guinea pigs and alters the expression of neurotrophins and neurotrophic factors. Our findings will aid mechanistic studies of neural dysfunction in eosinophilic esophagitis.

The growth inhibitory effects of garlic polysaccharide (GPS) combined with cis -dichlorodiamine platinum (DDP) on human HepG2 cells were evaluated in this paper. Our results showed that GPS, with a molecular weight of 2,714.94 Da, was isolated by precipitation and purification, and composed of glucose, galactose, xylose and arabinose with a relative molar ratio of 4.6: 0.8: 0.9: 0.5. GPS inhibited the proliferation of HepG2 cells. While, compared with GPS and DDP treated alone, the combination agents enhanced significantly growth inhibitory effect on HepG2 cells in a time- and dose-dependent manner. Propidium iodide/annexin V staining analyzed by flow cytometry demonstrated that GPS enhanced the cytotoxic effects of chemotherapeutic drug DDP on tumor cells. Cell cycle arrest of HepG2 treated with the combined compounds occurred in S/G2 phase. Western blot analysis revealed that the GPS can up-regulate the pro-apoptotic protein Bax. Our research provides additional evidence to develop combination therapies of GPS and DDP for human hepatic tumor and a potential way to decrease the side effect of DDP by combining GPS treatment.