Band 10, Heft 1

Some insect herbivores can regulate their nourishment intake by different feeding behaviour. This mechanism allows them to persist with utilising different food resources according to the composition of the vegetation within their habitats. Using a two-choice experiment, we analysed foraging behaviour in females of the tree-dwelling bush-cricket Barbitistes constrictus (Orthoptera), which originated from two different forest habitats, spruce and beech forest. We found that individuals from the spruce forest mainly foraged on needle tips, and thus they nibbled more needles per day than individuals from the beech forest (medians 106.0 vs. 42.5; p < 0.0001). However, when the contents of droppings were dissected, the volume of consumed spruce was similar in both groups of bush-crickets (median > 90%), which is explained by the different feeding techniques of bush-crickets from different habitats. We propose possible scenarios for bush-cricket feeding adaptations to the deleterious effects of the host plant chemical compounds serving as a plant defence against herbivores.

Human aggression/impulsivity-related traits are influenced by complex genetic and non-genetic factors. The aggression/anxiety relationship is controlled by highly conserved brain regions including the amygdala, hypothalamus and periaqueductal gray of the midbrain, which is responsible for neural circuits triggering defensive, aggressive, or avoidant behavioral models. The social behavior network consists of the medial amygdala, medial hypothalamus and periaqueductal gray, and it positively modulates reactive aggression. An important role in the incidence of aggressive behavior is played by secreted factors such as testosterone, glucocorticoids, pheromones, as well as by expression of genes such as neuroligin-2, monoamine oxidase A, serotonin transporters, etc. The authors deliberate whether aggression in sport is advantageous (or even indispensable), or to what extent it can hamper attainment of sport success. Methods of reducing and inhibiting expression of aggression in athletes are indicated.

In order to provide molecular characteristics of wild pear ( P. pyraster ) resources, six populations (192 accessions) from different regions of Poland were investigated with 17 SSR loci. Each of the SSR loci used was polymorphic, with a mean of 19.5 alleles per locus and a mean PIC of 0.806. Both the high heterozygosity (Ho = 0.751) and low Fis (0.007) indicated that the wild pear populations maintain a relatively high level of diversity, while the mean F index of 0.039 and the number of migrants per generation (Nm = 6.996) revealed a high gene flow and weak inter-population differentiation. AMOVA analysis located polymorphisms mainly within populations (96%). Genetic relations between populations did not show correlations with geographical distances. The dispersal influence of gene flow could be the reason of the disrupted relationship within populations and the low interpopulation differentiation. We did not find any evidence to support the hypothesis about influence of interspecies hybridization with pear cultivars on the level of wild pear population diversity.

Conductometric assessment of electrolyte leakage from spring barley plant organs subjected to higher cadmium concentrations 10 -2 – 10 -4 mol l -1 revealed serious tissue damage. Therefore plants were grown under controlled environment conditions in a phytotron in Knop´s nutrient solution in two variants – the control variant without cadmium and experimental variant treated during the growth of plants with CdCl2 solution of 10 -5 mol l -1 . Barley plants were then separated into roots, residual caryopses, leaf bases, and leaves. Lyophilized samples were analyzed for the uptake and redistribution of cadmium, calcium, copper, iron, zinc, and manganese in plant organs by spectrometric methods. Cadmium caused significant decrease of calcium and manganese levels in all analyzed plant parts indicating that cadmium was a distinctive antagonist to intake of these metals. Cadmium treatment also lowered amounts of zinc and iron in roots. An opposite trend was characteristic for Cu, where Cd stress caused significant Cu increase and retention in roots and significant decrease in leaves. The combined usage of the electroanalytical technique - conductometry and spectrometric methods (ICP-OES, AAS) proved to be a useful tool for monitoring of tissue damage and intake and transfer of cadmium and macro- and microelements into barley organs. Graphical Abstract

Pholiotina cyanopus was collected from wood chips and other woody remnants of undetermined tree species. Its basidiomata were found in June within the area of closed sawmill in the central part of Żywiec city (SW Poland). Description and illustration of Ph. cyanopus based on Polish specimens are provided and its ecology, general distribution and comparison with similar taxa – Pholiotina smithii, Pholiotina sulcatipes , and others are discussed as well. The identity of the active compounds of Ph. cyanopus was additionally determined. Liquid chromatography–mass spectrometry (LC-MS) data sets were obtained to support the occurrence of psilocybin and its analogues – psilocin, baeocystin, norbaeocystin, and aeruginascin in air-dried basidiomata of the species. The content of psilocybin was found to be high (0.90±0.08% of dry weight), besides, analysed samples contained lower concentrations of psilocin (0.17±0.01%), and baeocystin (0.16±0.01%). Additionally, the chemical analysis revealed small amounts of norbaeocystin (0.053±0.004%) and aeruginascin (0.011±0.0007%) for the first time in the species.

Cyanobacterial harmful algal blooms (or CyanoHABs) represent one of the most conspicuous waterborne microbial hazards. The characterization of the bloom communities remains problematic because the cyanobacterial taxonomy of certain genera has not yet been resolved. In this study, 29 planktic and benthic cyanobacterial strains were isolated from freshwaters located in Greece. The strains were assigned to the genera Chroococcus, Microcystis, Synechococcus, Jaaginema, Limnothrix, Pseudanabaena, Anabaena , and Calothrix and screened for the production of the cyanotoxins microcystins (MCs), cylindrospermopsins (CYNs), and saxitoxins (STXs) using molecular (PCR amplification of seven genes implicated in cyanotoxin biosynthesis) and immunological (ELISA) methods. This study presents, for the first time, a cyanobacteria culture collection from Greece, thus providing missing study material for the understanding of bloom formation and cyanotoxin production in the Mediterranean and for the polyphasic characterization of important components of the phytoplankton. The combined use of molecular and immunochemical methods allowed the identification of MC producing strains, but further data are needed for CYNand STX-producing cyanobacteria. The high percentage of MC-producing Microcystis strains in the urban Lakes Kastoria and Pamvotis, frequently used for agriculture irrigation, fishing and recreation, highlights the potential risk for human health.

The effect of environmental factors on bacterial and actinobacterial communities was assessed to predict microbial community structure in natural gradients. Bacterial and actinobacterial communities were studied at four sites differing in vegetation and water regime: creek sediment, wet meadow, dry meadow and deciduous forest located in a shallow valley. The vegetation structure was assessed by phytocoenological releves. T-RFLP and quantitative PCR were used to determine community composition and abundances. Significant relationships between bacterial community structure and selected soil traits at sites located relatively close to each other (within 200 m) were demonstrated. Both the quantity and structure of bacterial communities were significantly influenced by organic matter content, soil moisture and pH. Bacterial diversity was higher in summer, while that of actinobacteria increased in winter. The Simpson’s evenness E was significantly correlated with soil organic matter content. Soil pH had the greatest influence on bacterial community structure showing higher within-site variability in summer than in winter.

The floristic structure, distribution and habitat requirements of a forest syntaxon, Populetum pamiricae ass. nova is described. The potential range of the Populetum pamiricae association comprises the mountains of eastern Tajikistan (Pamir Alai Mts), Kashgaria in western China (Kunlun Shan Mts) and north-eastern Afghanistan (Hindu Kush Mts) in Middle Asia. The biotope of this community comprises of high mountain river valleys situated at elevations between 2,500 and 3,200 m above sea level. Phytocoenoses of the Populetum pamiricae association are characterized by the apparent domination in the tree layer of a diagnostic species, Populus pamirica. The tree stand has a mean cover around 55%. The layer of shrubs consists primarily of the willow including Salix linearifolia and S. wilhelmsiana, and Hippophaë rhamnoides, Rosa huntica and Lonicera stenantha. The herb layer is less developed and consists of species typical for Populetea laurifolio-suaveolentis; however, in juvenile stages of the community, taxa from riverbed vegetation (i.e. Salicion eleagni) also frequently occur. Populetum pamiricae grows in areas of flood inundations caused mainly by spring thaw where the humus layer in soils is very thin or even completely absent. The forest association of Populetum pamiricae ass. nova is one of the rarest and most rapidly disappearing wood communities in Tajikistan. High conservation priority and special management is recomended for this association.

In the midst of an obesity epidemic in humans, diet induced obesity studies in rodents are fundamental to unravel the complex mechanisms underlying this disease, ultimately resulting in the identification of new preventative and therapeutic strategies. The current study was designed to determine if high throughput multiobject CT scanning was capable of providing precise quantification of adipose tissue in C57BL/6 mice when benchmarked to the gold standard method for evaluating fat mass (freeze drying). We report a strong correlation between body weight alone and fat percentage in our mouse cohort (20 g-40 g, r = 0.95). The gonadal fat depot was identified as the most accurate single predictor of total fat mass (r = 0.931). Importantly, we observed a high positive correlation between both live tissue weight and dissected adipose tissue when correlated to CT predictions (r ≥ 0.862), suggesting CT can accurately be used to predict total fat mass/percentage and non-fat mass/percentage in our cohort. We conclude that the use of multi-object in vivo CT fat quantification is cost effective, accurate and minimally invasive technique in the genetic manipulation era to exploit lean/obese genes in the study of diet induced obesity, allowing longitudinal studies to be completed in a high throughput manner.

Background: The extracts of Allium cepa are known for their medical use: antioxidant, antiinflammatory, antimicrobial, fibrinolytic and antiplatelet properties. Our study aims to establish, using in vitro tests, the antiplatelet and antioxidant character, the link between them and the extract acidity, from seven varieties of A. cepa. Methodology: The qualitative and quantitative presence of polyphenols and anthocyanins in the extracts was determined using UV-Vis and HPLC. Quantitative determination of the thiosulfinates compounds was calculated using their reaction with 4-mercaptopyridine. Antioxidant character was determined using 3 methods (FC, DPPH and TEAC), and antiplatelet effect was measured by in vitro tests on platelet rich plasma obtained from human blood. Principal Findings/Results: The white variety of A. cepa has the most alkaline pH, the largest amount of thiosulfinate compounds and the most powerful antiplatelet effect, but a very small amount of flavonoids and an antioxidant effect almost nonexistent, in contrast with red variety of A. cepa which is the opposite. Conclusions/Significance: The white variety of A. cepa had very high anitiplatelet activity suggesting the potential use of A. cepa extract in treating cardiovascular diseases.

The objectives of our survey were: to analyze the structure of macroinvetebrate communities in mountain streams in national parks and Biosphere Reserves (Poland, the Slovak Republic), to determine the environmental factors that influence the structure of macroinvertebrate communities and to assess the stream habitats including the bank and channel features, any modifications, land use and channel vegetation. Our results showed that in addition to the conductivity, the altitude, stream gradient and the values of the HQA index that reflected more natural features in the channel and river corridor were most important. The River Habitat Survey (RHS) method reflects not only the morphology but also the relationships between habitat features and the structure of macroinvertebrate communities and it provides a more holistic approach to assessing the health condition of stream ecosystems. Headwater streams support unique macroinvertebrate taxa that are found nowhere else in a catchment and may also constitute refuges for in-stream biota. Some of the least water pollution-tolerant macroinvertebrate taxa were recorded.

Oat (Avena sativa L.) has recently gained importance due to the discovery of a variety of health benefits and new opportunities of use. There is no efficient protocol for the production of oat doubled haploid (DH) lines. The aim of this study was to increase the efficiency of obtaining DHs of oat by the wide crossing method. The study was performed on five oat genotypes. We have compared the induction of embryos after pollination with maize, sorghum and millet pollen as well as the development of haploid embryos isolated 2, 3 and 4 weeks after pollination and cultivated on media with different sugar content. Haploid plants were treated with colchicine after or before acclimation to natural conditions. Of the three types of pollen used, the largest number of haploid embryos was obtained using maize pollen. Three weeks after pollination was the most suitable time for the isolation and cultivation of the embryos. The most efficient medium enabling the development of embryos and conversion to plants was 190-2 containing 9% of maltose. Colchicine treatment of acclimated plants provided high survival rate.

Water blooms dominated by cyanobacteria are capable of producing hepatotoxins known as microcystins. These toxins are dangerous to people and to the environment. Therefore, for a better understanding of the biological termination of this increasingly common phenomenon, bacteria with the potential to degrade cyanobacteria-derived hepatotoxins and the degradative activity of culturable bacteria were studied. Based on the presence of the mlrA gene, bacteria with a homology to the Sphingopyxis and Stenotrophomonas genera were identified as those presenting potential for microcystins degradation directly in the water samples from the Sulejów Reservoir (SU, Central Poland). However, this biodegrading potential has not been confirmed in in vitro experiments. The degrading activity of the culturable isolates from the water studied was determined in more than 30 bacterial mixes. An analysis of the biodegradation of the microcystin-LR (MC-LR) together with an analysis of the phylogenetic affiliation of bacteria demonstrated for the first time that bacteria homologous to the Aeromonas genus were able to degrade the mentioned hepatotoxin, although the mlrA gene was not amplified. The maximal removal efficiency of MC-LR was 48%. This study demonstrates a new aspect of interactions between the microcystin-containing cyanobacteria and bacteria from the Aeromonas genus.

Down syndrome (DS) is characterised by intellectual disability and is caused by trisomy 21. Apoptosis is a programmed cell death process and is involved in neurodegenerative diseases such as Alzheimer. People with DS can develop some traits of Alzheimer disease at an earlier age than subjects without trisomy 21. The leucine zipper, down regulated in cancer 1 (LDOC1) appears to be involved in the apoptotic pathways. The aim of the present work was to detect the presence of intracellular synthesis of LDOC1 protein and LDOC1 mRNA in fibroblast cultures from DS subjects. The western blot shows the presence of LDOC1 protein in fibroblasts of DS subjects but no evidence of LDOC1 protein in fibroblasts of normal subjects. LDOC1 gene mRNA expression is increased in fibroblasts from DS subjects compared to fibroblasts from normal subjects. The data obtained from this study strengthen the hypothesis that the over-expression of LDOC1 gene could play a role in determining the phenotype of individuals with DS but does not exclude that this results from apoptotic mechanisms.

Potato virus X (PVX) is widely used as a peptide presentation system in plant biotechnology, mostly for transient expression of desired peptides fused to the N-terminus of its capsid protein (CP). Here we describe testing/investigation of new positions for peptide presentation based on seven putative surface loops of PVX CP. We performed bacterial expression of fourteen different PVX CPs modified by the insertion of the epitope derived from the E7 oncoprotein (E7 epitope) of Human papillomavirus type 16 fused with His tag. The expression from vector pMPM-A4Ω in Escherichia coli MC1061 was performed to evaluate the capacity of the PVX CP platform to tolerate the insertion of E7 epitope fused with His tag in the seven putative surface loops. The immunological characteristics of expressed epitopes were assessed by Western blot using both anti-PVX CP and anti-His antibodies and by immunoelectron microscopy.

In this study, an efficient and reproducible plant regeneration system for lotus (Nelumbo nucifera Gaertn.) was established using shoot apical meristems from the buds and one-week-old aseptically germinated embryos as explants. Multiple shoot clumps were induced on Murashige and Skoog (MS) basal medium supplemented with various combinations of N6-Benzylaminopurine (6-BA) and α-Naphthalene acetic acid (NAA). The maximum response was obtained with 2.22 μM 6-BA, and produced 21.33 shoots per explant after four weeks. After five subcultures, multiple shoot clumps were transferred to MS basal medium supplemented with various combinations of 3-Indolebutyric acid (IBA), NAA and sucrose for root induction. After four weeks, plantlets with well-developed roots were achieved on MS basal medium supplemented with 0.54 μM NAA and 30 g/L sucrose with 100% rooting rate. The successfully acclimated plantlets were transferred to pots with the addition of 2 g/L KMnO4 into the soils, and finally fertile plants with much bigger leaves were obtained in the greenhouse. The survival rate was 97.33%.

Symphyotrichum ciliatum was reported as invasive in Romania in the early 1970s. In this study we have analysed its invasion history, current distribution in Romania, habitat preferences and the associated plant communities. Data recorded during our recent field work, as well as from herbaria and previously published papers, were used in the analysis. This data confirms its invasive character. Given the abrupt increase in the slope of the invasion curve during the last decade, we can assume that S. ciliatum is still far from reaching the saturation phase of its invasion in Romania. Habitats invaded by this species are mostly anthropogenic herb stands associated with rail and road transport networks, and the continental inland salt steppes. The classical methodology of the Zürich- Montpellier school was used for the field phytosociological research and the classification of the investigated communities was done using the deductive method. The phytocoenoses dominated by S. ciliatum were classified as derivate communities. The description of these communities included data about plant composition, synecology and distribution in the study area.

The paper presents the impact of periodic soil drought on physiological parameters important for bioproductivity of mountain beech populations. The investigated forest population was located near Fruška gora mountain peak, where water runs off quickly, and consequently lack of soil humidity develops very often. Decreasing trends of photosynthesis, transpiration, water use efficiency and stomatal conductance (gs) during the growing season were evident, in correlation with a shortage of precipitation. Diurnally, photosynthesis of beech leaves showed rhythmical changes. It was the most intensive in the morning, then decreased between noon and 1 pm, and increased again during early afternoon. High leaf temperature and water deficit in the deeper soil layers caused a decreasing trend in photosynthesis and daily rhythmic changes of the transpiration rate and water use efficiency. Although surface soil water capacity did not show a significant decreasing trend from July to September, a lack of precipitation was observed, which probably caused a lack of moisture in deeper soil layers, resulting in a decline in photosynthesis and transpiration. Physiological status, linked to primary photosynthetic productivity of forests, could be a significant indicator of environmental conditions and trends in climate changes.

Non-coding rDNA spacers (IGS) can vary substantially in size due to differences in the number of repetitive elements among closely related species. Three pairs of universal primers were used in this study for the amplification of non-coding regions of ribosomal (rRNA) IGS. The amplified IGS products obtained from 19 Secale accessions, which included both cultivated and noncultivated rye and which represented three species and four subspecies of the genus Secale, showed a high level of polymorphism. The PCR results were characterized by multiple bands (1-6), different sizes (750bp-3250bp) and 100% polymorphism. Cluster analysis using the neighborjoining method based on the Dice’s coefficient of genetic similarity showed a division of the studied species into two similarity groups. All the studied Secale cereale ssp. cereale were found to belong to the same similarity group. The variation in the size of the IGS among the species which was detected in this study could be due to dissimilarity between the sequences of their respective repetitive elements or between their tandem repeat numbers. The highly interspecific polymorphisms for the rDNA IGS regions suggested that IGS might be a useful molecular marker in studies of Secale species.

In the Old-Polish Industrial Region (Southern Poland, Central Europe), specific remnants of former iron-ore extraction occur. These interesting post-mining habitats influence the plant cover which emerges on them. In this paper, I analyzed the impact of the former iron-ore mining on the increase of plant species richness in forest communities. I analyzed the floristic composition of 100 phytosociological relevés (plots) made in the mining areas, their surroundings and 100 associated soil samples. I found that within the island communities developing in the former mining fields, there was a considerable increase in the number of species of native flora (in comparison with communities of nontransformed areas). This increase in species richness is caused chiefly by the increased proportion of species characteristic of the syntaxa from the Querco-Fagetea class – species exclusively attached to the mesophilous forest communities developing in gob piles whose limits are marked by the material extracted and scattered around in ancient times. My research suggested that in this case, mining activity exerted a positive influence on the biodiversity in forested areas, improving the quality of soils and creating mesophilous forest communities growing on remnants of former iron-ore extraction, which are specific islands among acidophilous forest communities growing in the non-transformed areas.

Tularemia is a debilitating febrile and potentially fatal zoonotic disease of humans and other vertebrates caused by the Gram-negative bacterium Francisella tularensis. The natural reservoirs are small rodents, hares, and possibly amoebas in water. The etiological agent, Francisella tularensis, is a non-spore forming, encapsulated, facultative intracellular bacterium, a member of the γ-Proteobacteria class of Gram-negative bacteria. Francisella tularensis is capable of invading and replicating within phagocytic as well as non-phagocytic cells and modulate inflammatory response. Infection by the pulmonary, dermal, or oral routes, respectively, results in pneumonic, ulceroglandular, or oropharyngeal tularemia. The highest mortality rates are associated with the pneumonic form of this disease. All members of Francisella tularensis species cause more or less severe disease Due to their abilities to be transmitted to humans via multiple routes and to be disseminated via biological aerosol that can cause the disease after inhalation of even an extremely low infectious dose, Francisella tularensis has been classified as a Category A bioterrorism agent. The current standard of care for tularemia is treatment with antibiotics, as this therapy is highly effective if used soon after infection, although it is not, however, absolutely effective in all cases.

Lactobacillus fermentum ME-3 (LfME-3) has been proven to synthesize and secrete glutathione. A regular use of the foods fermented by it has shown a favourable influence on human lipid profiles and several antioxidant parameters. We administered the LfME-3-fermented kefir for 14 days to 43 human subjects and evaluated their serum with MALDI-TOF mass spectrometer at the beginning and end of the test period. We found an increase of the peak at m/z 308 (corresponding to glutathione) and a new peak at m/z 1467.

This study has assessed the protective efficacy of Silybum marianum seed oil (SMSO) in the context of CCl 4 -induced injury and oxidative stress in murine liver. Based on the GC-MS analysis, linoleic and stearic acids, tocopherol, ascorbic acid 2,6 dihexadecanoate and other constituents were identified in SMSO. Swiss mice received oral doses of SMSO daily for 21 days (10 g/kg b.w.) and subsequently injected i.p. with CCl 4 (50% v/v in olive oil; 1 ml/kg) on the 22nd day. CCl 4 administration induced an elevation of serum amino- and glutamyl transferases activities and an increased peroxidation, as well as a decrease of SOD, CAT, GPx, GR and GST activities in liver. SMSO successfully prevented oxidative stress and restored the biochemical parameters, hepatic architecture and expression of TNF-alpha. These findings suggest that SMSO was effective in counteracting the damaging effects of CCl 4 -induced injury in hepatocytes, probably due to its inherent antioxidant properties.

Stonefly samples were collected from disturbed and undisturbed tributaries of the Tatra Mountains streams (the West Carpathians). In the autumn, at stable low discharge, the total density of stoneflies was significantly higher in the undisturbed streams. Microhabitats such as macrolithal (boulders), mesolithal (stones) and moss had higher stonefly density. Taxa of different species or genera have different demands for microhabitats. Very narrow spatial niches were found for the genera Rhabdiopteryx, Protonemura and Perlodes . The spatial niche overlap was low between the genera Rhabdiopteryx and Brachyptera , but was high between species of the Protonemura and Leucra genera. The highest biodiversity of stoneflies was on coarser substrata (except boulders) and moss, the lowest biodiversity was on the finer substrata. Among the organic substrata, a significantly lower coefficient of stonefly α-diversity variation was recorded in mosses compared to submerged wood and roots.

The influence of different fertilization treatments on soil biological characteristics and microbial community structure was investigated. Soil samples were taken from a long-term field experiment which was conducted to explore the effect of eight treatments: control (non-fertilized), NPK (nitrogen, phosphorus and potassium), FYM (farmyard manure), FYM + NPK, CSl + St (cattle slurry + straw), CSl + St + NPK, CSl, CSl + NPK. The highest values of invertase activity were found in treatment fertilized with farmyard manure combined mineral fertilizers, similarly to total N content, activity of urease, and C of microbial biomass. Dehydrogenase activity was lower in all treatments with mineral fertilization. Bacterial and actinobacterial T-RFLP profiles of 16S rRNA genes showed similar patterns in response to eight fertilization treatments. In both, the communities formed distinct groups, which were separated by organic fertilization i.e. cattle slurry and straw amendments along the x axes and by NPK amendments along the y axes using the Sammon’s method of multidimensional scaling. Significant correlations were determined in several situations related to diversity: between invertase and 1/Db (dominance, bacteria), 1/Da (dominance, actinobacteria) and between T-RFLP profiles of actinobacteria, nitrogen and organic carbon content and bacterial E (evenness).

The aim of this study was to evaluate the influence of phenolic, flavonoid, and anthraquinones from sequential extracts (n-hexane, dichloromethane, ethyl acetate, and ethanol) of Embothrium coccineum leaves on the antioxidant capacity, cell viability, and toxicity of the same, in order to find possible sources for novel antioxidants for food and pharmaceutical formulations. Antioxidant potential of sequential extracts was analyzed by five different assays: 2,2-diphenyl-1- picrylhydrazyl radical scavenging activity assay (DPPH), hydrogen peroxide scavenging activity (H2O2), oxygen radical absorbance capacity (ORAC), ferric reducing antioxidant power (FRAP), and total reactive antioxidant potential (TRAP). An in vitro growth inhibition assay was performed using sulphorhodamine dye to quantify cell viability, while an in vivo brine shrimp lethality test was used to quantify toxicity. The dichloromethane extract has a greater efficiency in scavenging free radicals, combined with low toxicity, and no effect exhibited on healthy cells, compared to observations for the other extracts tested. Further research is in progress to identify and separate the active compounds of active extracts and investigate the protective effect of extracts on human dermal fibroblast injury induced by hydrogen peroxide.

As a target of insect growth regulators, the ecdysone receptor complex binds ecdysone agonists to regulate gene transcription, development, and growth. To understand the response of Lymantria dispar to methoxyfenozide, we studied receptor complex transcription profiles following larval exposure to methoxyfenozide, and observed important variations amongst post-embryonic stages. After the exposure of 2nd and 3rd instar larvae to LC5 and LC20 methoxyfenozide for 72 h, the transcription levels of ecdysone receptor complex genes were mainly upregulated in 2nd instar larvae, but downregulated in 3rd instar larvae. These results provide expression characteristics of the ecdysone receptor complex in L. dispar larvae after methoxyfenozide exposure.

Objective: Ileal transposition procedure (IT), in combination with sleeve gastrectomy, is widely used to induce diabetes remission and to control related metabolic abnormalities. A transposition of a long segment of distal ileum in obese Zucker rats improved glucose tolerance 6 months after IT. The premise of our study was to to examine the long - term effects of ileum transposition on the liver glycolytic enzymes content in a euglycemic group of operated Zucker rats. Methods: Twenty male Zucker rats underwent either the transposition of 50% distal ileum or a sham surgery. Six months after surgery, liver tissue concentrations of glycogen synthase kinase alpha (GSK-3α), glucose 6-phosphatase (G6PC), glycogen phosphorylase (PYGM) and phosphofructokinase (PFK) and HSP70 were assessed by immunoenzymatic methods. Results: HSP70 values were significantly higher in the IT group compared to SHAM. G6PC liver concentrations in the IT group were almost 1.45-fold lower than in the SHAM operated rats. Statistical analyses (F-test) showed HSP70 levels were significantly related to caveolin-1and SHAM group. Conclusions: Lowered glycolytic enzyme concentrations assessed in the liver suggest positive effects on glucose metabolism in long-term observations.

Environmental scanning electron microscopy enables the investigation of uncoated pine early embryogenic tissue samples in situ. The samples were examined under low vacuum conditions (air pressure 550 Pa) at a temperature of around -18°C by the AQUASEM II noncommercial environmental scanning electron microscope. The native extracellular matrix surface network was imaged by the environmental scanning electron microscope and in dark field mode of the optical microscope too. The backscattered electron detector disclosed brightness loci in the cells of early embryogenic culture. This work shows images of native pine embryogenic tissues. The continuity of extracellular matrix with structural integrity of plant organism is discussed.

BACKGROUND: “VITALPLANT” medicinal plant mixture, aimed at body weight regulation and metabolism enhancement was formulated in order to be incorporated as an ingredient in functional food formulations on the basis of known biological activity of the alder buckthorn bark, peppermint, caraway and parsley. METHODOLOGY: The body weight-lowering potential, hypolipidemic effects and antioxidant activity of “Vitalplant” mixture were examined on five groups of male Wistar rats on different food regimes for 14 weeks. RESULTS: “Vitalplant” mixture supplementation caused an increase in food intake, but induced a decrease in body weight gain. The lipid status and atherogenic index (AI) were significantly improved in “Vitalplant” mixture supplemented hyperlipidemic (HF) groups in comparison to HF group. “Vitalplant” mixture was able to influence the activity of antioxidant enzymes and to protect erythrocytes of the rats fed the HF diet from oxidative damage, which was indicated by significantly lower malondialdehyde (MDA) levels in “Vitalplant”- HF groups in comparison to the HF group. CONCLUSION: “Vitalplant” mixture seemed to be beneficial for the reduction of body weight and improvement of antioxidant status of the erythrocytes, and its antihyperlipidemic property was highly active for enhancing the profile of plasma lipids in rats.

Glutamine synthetase (GS) plays a fundamental role in nitrogen metabolism in higher plants. Three BnGS genes have first been isolated: one gene encoding plastid GS (BnGS2) and two encoding cytosolic GS (BnGS1-1 and BnGS1-2) in ramie. Based on a sequence analysis and phylogenetic study, three BnGS sequences were classified into three distinct sub-families. The phylogenetic analysis showed that BnGS2 and BnGS1-2 were closely related to those of legumes, alfalfa (Medicago sativa), soybean (Glycine max) and bean (Phaseolus vulgaris). The BnGS gene expression patterns revealed that each gene exhibited similar organ specificity, but distinct transcript intensity during different vegetative processes. The relatively abundant expression of BnGS1-1 and BnGS2 at specific organs during different vegetative processes indicates that they have critical roles in nitrogen uptake and assimilation relating to forage and growth characteristics. The BnGS1-2 mRNA levels were remarkably upregulated in the phloem, xylem and stems during the fiber development stage, suggesting a correlation with fiber development. Therefore, the non-overlapping transcript intensity of BnGS genes in different tissues regulates ramie growth and development during different vegetative processes.

The cut chrysanthemum (Chrysanthemum ×morifolium Ramat.) is one of the most recognized cut flowers in the world. Light quality significantly affects the development and growth of plants. To conduct an in-depth investigation of the effects of light quality on the growth and development of cut chrysanthemums, seedlings of the cut chrysanthemum cultivar ‘Riqietaohong’ were cultured under four different light emitting diode (LED) systems: red (R), blue (B), red : blue = 2:1 (RB) and blue : red = 2:1 (BR). A fluorescent lamp was used for control group lighting. The Duncan multiple comparisons analysis showed that the B light promoted stem and leaf development of seedlings but inhibited photosynthesis. The R light stimulated their rooting but inhibited the accumulation of biomass. RB improved transpiration. Moreover, a comprehensive evaluation system that combined the analytic hierarchy process (AHP) method with the technique for order preference by similarity to an ideal solution (TOPSIS) method indicated that the photosynthesis index, transpiration index and biomass index were the most important indicators for the growth and development of cut chrysanthemums, and RB was the best LED system for cut chrysanthemums. The present study provides references for the lower energy consumption and industrial annual production of cut chrysanthemums.

RNase H2 consists of three subunits (H2A, H2B and H2C) and is involved in the hydrolysis of RNA/DNA hybrids. The GRG motif in RNase H2 is highly conserved and recognizes the ribonucleotides misincorporated into dsDNA. The mutant G37S in the GRG motif of human RNase H2A was found to be correlated with Aicardi-Goutièressyndrome (AGS). In this study, 4 mutants (G37S, G37A, R38A and G39A) were prepared and their biochemical properties of secondary structure, activity and binding affinity with substrate were studied in order to explore the function of the GRG motif. The activity assay showed that the mutations resulted in significantly decreased RNase activity. The binding efficiency assay demonstrated that binding affinities between 4 mutants and 1-ribo FAMlabeled substrate were significantly weakened. These results all indicated that the GRG motif contributes directly to substrate binding and is closely related to the enzyme activity.

Little has been published about the occurrence, speciesidentification, andpathogenicpotentialofcoagulasenegative staphylococci (CoNS) present in drinking water. In this study, ten species were identified among 57 isolates of staphylococci from 756 samples of chlorinated drinking water taken from public distribution networks in the Slovak Republic. S. warneri (37%), S. haemolyticus (23%), and S. saprophyticus ssp. saprophyticus (14%) were identified most frequently. Isolates did not produce coagulase, DNase, or hyaluronidase; production of gelatinase and lecithinase was observed in 28 and 22 isolates, respectively. Genetically encoded ability for production of enterotoxin SED was revealed in two isolates. Among ten antibiotics tested, resistance to ampicillin (66.7%), penicillin (64.9%), and erythromycin (57.9%) were observed most frequently. Resistance to gentamicin, vancomycin, or clindamycin was not confirmed. Production of β-lactamase was observed in 64.9% of isolates. Fourty-two isolates were resistant to two or more antibiotics tested, and eight isolates showed multiresistance. The presence of mecA gene was confirmed in 8 isolates, while PBP2a was revealed in 7. Two isolates of S. epidermidis were identified as methicillin-resistant (MRSE). The results demonstrate that CoNS in chlorinated drinking water may possess virulence factors and show resistance to various antibiotics. Therefore, their pathogenic potential should not be ignored.

Tobacco (Nicotiana tabacum L.) is an economically important and relatively drought-tolerant crop grown around the world. However, the molecular regulatory mechanisms involved in tobacco root development in response to drought stress are not wellknown. To gain insight into the transcriptome dynamics associated with drought resistance, genome-wide gene expression profiling of roots from a tobacco cultivar (Honghua Dajinyuan, a major flue-cured tobacco cultivar in Southwest China) under 20% PEG6000 treatment for 0, 6 h and 48 h were conducted using Solexa sequencing (Illumina Inc., San Diego, CA, USA). Over five million tags were generated from tobacco roots, including 229,344, 221,248 and 242,065 clean tags in three libraries, respectively. The most differentially expressed tags, with either log2FC > 2.0 for up-regulated genes or log2FC < -2.0 for down regulated genes (p < 0.001), were analyzed further. In comparison to the control, 1476 up-regulated and 1574 down-regulated differentially expressed genes (DEGs) were identified, except for unknown transcripts, which were grouped into 43 functional categories involved in seven significant pathways. The most enriched categories were those that were populated by transcripts involved in metabolism, signal transduction and cellular transport. Many genes and/or biological pathways were found to be common among the three libraries, for example, genes participating in transport, stress response, auxin transport and signaling, etc. Next, the expression patterns of 12 genes were assessed with quantitative real-time PCR, the results of which agreed with the Solexa analysis. In conclusion, we revealed complex changes in the transcriptome during tobacco root development related to drought resistance, and provided a comprehensive set of data that is essential to understanding the molecular regulatory mechanisms involved. These data may prove valuable in future studies of the molecular mechanisms regulating root development in response to drought stress in tobacco and other plants.

A variable response of Prunus avium L. seeds to desiccation and storage in liquid nitrogen (LN) has been reported in the literature. The majority of these experiments were conducted on initially dried seeds. The desiccation and LN exposure tolerance of fresh P. avium seeds is unknown. In the present study, fresh seeds were used to determine seed response to desiccation and cryopreservation. Desiccation of seeds from a moisture content (MC) of 19.7-20.2% to 10.1-10.9% or 07.6-8.5% reduced seedling emergence from approximately 73 to 19 and 16% for first provenance; and from approximately 89 to 10-12% for second provenance of seeds. After exposure to LN, seeds had the highest seedling emergence when seed MC was the highest (19.7 and 20.2%, respectively) prior to cryostorage. Results indicated that P. avium seeds should be classified as intermediate. For cryopreservation in seed banks, we recommend that seeds be dried directly after extraction from fruits in the range of 16.8-20.2% of MC (0.21-0.25 g·g –1 of WC) and directly immersed in LN in tightly closed cryovials.

Background Metabolic syndrome (MS) has a great impact on cardiovascular mortality and morbidity. Our aim was to investigate the association of MS with some oxidant and antioxidant markers, including pro-and antioxidant status of peripheral blood mononuclear cells (PBMC) in newly diagnosed type 2 diabetic mellitus patients (ND-T2D). Methods 219 ND-T2D and 88 healthy subjects were divided in two groups according to the absence or presence of MS. Anthropometric measurements, routine blood tests, total oxidant status (TOS), total antioxidant status (TAS) and ELISA measurements were included. The PBMC capacity to release free radicals and to neutralize them was also determined by measuring the respiratory burst (RB) together with the lactonase activity of the intracellular antioxidant enzyme paraoxonase 2 (PON2). Results Comparing ND-T2D MS+ with those MS- the RB of the PBMC was significantly higher (p<0.05) while lactonase PON2 enzymatic activity was decreased (p < 0.001). A negative correlation of RB was found with TAS (r = -0.416, p < 0.05). PON2 was also negatively correlated with glycaemia (r = -0.275, p < 0.001), HbA1c (r = -0.308, p < 0.001), weight (r = -0.183; p < 0.05), waist circumference (r = -0.353, p < 0.001) and body mass index (r = -0.290, p < 0.001). Conclusion PON2 lactonase activity is negatively associated with anthropometric markers in ND-T2D with MS.

Thaumetopoea pityocampa, the winter pine processionary moth, is one of the most important pests of pine trees in the Mediterranean region. To learn more about the refugial areas within the greater refugium of Greece, samples from 15 local populations were collected. Analysis of the Cytochrome Oxidase I region identified 15 haplotypes. One strongly supported clade was found, separating the Aegean island of Lesvos from the mainland populations. Mdiv analysis showed that this clade diverged from the general clade 155,000 y.a., suggesting the sea level changes during the Quaternary Period as a possible reason for its seclusion. Additionally, comparisons with similar studies in this region revealed a possible colonization of the eastern Aegean islands from the Turkish mainland rather than from Greece. However, no strong isolation-by-distance events were detected among the mainland populations, which could be attributed to the joint effect of regular gene flow and the lack of insurmountable geographic barriers. Finally, regarding population structure, Bayesian analysis as well as neutrality tests pointed towards an ongoing population expansion which verifies the potential invasiveness of this pest species, something that will have to be dealt with under the influence of climate change.

Background: The effects of two lipidlowering drugs, simvastatin and fenofibrate on osteoporosis in the femurs of healthy and ovariectomized female rats were investigated quantitatively by histological images and 1 H NMR relaxometry. Methodology: The intertrabecular cavities percentage was estimated from numerically cleaned histological images which initially present both the trabecular bone architecture and bone marrow filling the intertrabecular pores. The eight weeks evolution of intertrabecular cavities percentage measured from histological images of randomly selected sections in ovariectomized rats femoral diaphysis is similar with the evolution of transverse relaxation time centre of gravity T 2,CG , a selected parameter to quantify the changes in intertrabecular cavities dimensions from onedimensional (1D) transverse relaxation time T 2 distributions. Results: The analysis of histological images recorded for non-ovariectomized rats show that the treatment with simvastatin and fenofibrate has a negative effect on the dimension of the femoral diaphysis trabecular bone. The analysis of 1D T 2 distributions show that the simvastatin and fenofibrate lipid-lowering drugs produce different effects on different particular sections of studied rat bones, i.e. proximal part of femoris, diaphysis or distal epiphysis. Conclusions: Finally, it is shown that the effects of the treatments are strongly dependent on the duration of treatment.

The signaling pathways that control intestinal development, regeneration and disease show a high degree of conservation between Drosophila and mammals. The gut epithelia of Drosophila provide protection against invasion of microorganisms through production of reactive oxygen species (ROS) and antimicrobial peptides (AMPs). Although Drosophila gut immunity has been extensively studied, the specific responses to Gram-positive bacteria, fungi and toxic compounds are not fully understood. To identify the physiological role of genes involved in host defense we studied Drosophila mutants in antifungal genes identified previously and tested their survival upon feeding with various pathogens and toxic compounds. The results showed that several mutants displayed decreased viability compared with wild-type flies, and the lower survival rates were attributed to morphological change and excessive cell death in mutant guts. Thus, we identified several new Drosophila genes (spen, jumeau, inv, DDB1 and shg) required for intestinal homeostasis or stress responses.

Notch signaling plays a key role in cell proliferation and differentiation, and is important in several biological processes, but its role in the chondrogenesis of human umbilical cord mesenchymal stem cells (UC-MSCs) is still unknown. N-[N-(3,5- difluorophenacetyl-L-alanyl)]-(S)-phenylglycinet-butyl ester (DAPT) is the inhibitor of Notch pathway. The aim of this study is to investgate the influence of DAPT on the chondrogenesis of UC-MSCs. In our study, UC-MSCs were isolated from human umbilical cord and their characteristics were identified. The UC-MSCs were induced to differentiate into chondrocytes in vitro and treated with 5 μM DAPT. Glycosaminoglycan (GAG) and collagen type II (COL-2A1) were analyzed qualitatively and quantitatively. The gene expression of Notch-1, Hes-1, GAG and COL-2A1 were analyzed by quantitative polymerase chain reaction (qPCR). The UC-MSCs separated from human umbilical cord, followed the characteristics of Mesenchymal Stem Cells (MSCs). The gene expression of Notch-1 and Hes-1 decreased after chondrogenic induction but the percentage in G1 period and the content of GAG and COL-2A1 increased. The expression of all tested Notch signaling and proliferation genes declined when 5 μM DAPT was added, also the content of GAG and COL-2A1 also decreased. Our study revealed that Notch signaling exists in UC-MSCs and it may remain the proliferative activity of UC-MSCs. Once the chondrogenesis begins, Notch signaling strength decline evidently. DAPT inhibits the chondrogenesis of UC-MSCs.

The present paper describes the antifungal activity of some plant extracts on the development of Fusarium oxysporum f.sp. lycopersici. The best extracts were selected to be tested as a phytofungicide to control crop diseases, with the ultimate goal of developing a green alternative to synthetic fungicides. Using the conidia germination assay, of the 24 plant extracts tested, 15 reduced conidia germination and 6 completely inhibited germination. Extracts of Rivina humulis, Brassica carinata, Brunfelsia calyicina, Salvia guaranitica and Punica granatum showed good antifungal activity. The relationship between total phenolic content (TPC) in each plant extract tested and the percentage of mycelial growth inhibition showed a significant correlation (R2 = 0.69), while no correlation was found between total flavonoid content (TFC) and percentage mycelial growth inhibition. Among all extracts tested, Punica granatum and Salvia guaranitica showed the best inhibitory effect against Fusarium oxysporum f.sp. lycopersici . Our results indicate that plant extracts with a good antifungal activity generally had a high level of total polyphenolic content and titratable acidity, and low values of pH.

The main function of the Immune System (IS) in metazoans is to protect them against invading microorganisms, which are considered from this perspective as competitors that reduce the availability of resources, cause tissue damage and essentially threaten their adaptability. This relationship is seen as a low intensity perpetual armed war, related to the hypothesis of the “Red Queen”. This view suggests that the metazoans’ IS has evolved under a selective pressure imposed by microorganisms of the microbiota, that whether infectious or not, have co-evolved with the host’s IS in an evolutionary symbiogenesis, with reciprocal interactions that have developed local immunity. The microbiota changes with age and communicates with the brain. The above mentioned implies that we are witnessing the birth of a new scientific discipline that could be termed microbial anthropology, and several perspectives that includes the change of our perception of health and established the fundamentals for the use of microorganisms as therapeutic agents. In order for these therapies to succeed considerable information must be available about our microbiota, such as changes during pregnancy, the effects of antibiotic use, new methods for in vitro cultivation and the host-commensal dynamics.

Plants of Heracleum sosnowskyi Manden. are rich in essential oil which might be of herbicidal potential. In this study, we examined 1) the content and chemical composition of H. sosnowskyi essential oil (EO) distilled from seeds and 2) the herbicidal effect of EO in a bioassay against germination of maize and five weed species. As a result, a 5.1% EO yield was obtained from the seeds of H. sosnowskyi. We identified sixty-two compounds of EO that constituted 96% of the total oil. Aliphatic esters were the main constituents of the EO, followed by aliphatic alcohols. The tested seeds showed different susceptibilities to the tested concentrations of EO (0.2 – 7.2 g L -1 ). The most susceptible were Bromus secalinus and Avena fatua, and the most resistant were Echinochloa crus-galli and maize. Kernels of maize germinated normally up to a 0.6 g L -1 dose of EO, and 20% of the seedlings were still germinating in the presence of 7.2 g L -1 of EO. These findings suggest that the EO of H. sosnowskyi and its main compounds should be studied further in soil conditions for its herbicidal properties against A. fatua and weeds in maize.

Alzheimer disease (AD) is responsible for the majority of elderly dementia cases. There are multiple mouse models of AD, however, none of them completely recapitulate human pathology. Moreover, it is even more difficult to imitate the onset of the disease in experimental models. In the current study we analyzed the influence of low amyloid level toxicity on the function of hippocampal neurons. Our assay is based on use of synthetic oligomeric amyloid beta peptides. These peptides were added to primary hippocampal cultures at a physiological nanomolar range to mimic early AD. Calcium imaging was used to evaluate the functionality of the assay. A low amyloid level toxicity assay could be useful for fundamental research as well as for testing newly developed AD drugs.

European cisco (Coregonus albula L.) is one of the most precious species of the European lake ichtyofauna, however, due to progressive eutrophication of water, the range of its occurrence has decreased. Deteriorating ecological conditions are the main cause of this decline in population, and most of the existing populations in lakes are maintained thanks to reintroduction. Thus, it is important to determine the genetic structure of the European cisco. The study involved PCR-RFLP-based genetic analysis of C. albula caught in 15 lakes in northern Poland, including four lakes located in national parks. The analysis covered 3 genes located in the mitochondrial DNA: ND1, ND3/4 and cytochrome b, as well as a control region (D-loop). The PCR product was digested with 4 endonucleases (RsaI, MspI, BsuI and HhaI) and the resulting haplotypes were grouped into combinations. Statistical analysis were then performed on these groups. Based on the genetic distance, a phenogram was constructed in which two groups could be distinguished. One group was represented by C. albula populations from most lakes in north-western Poland, including the three protected lakes. The other group consisted of the European cisco population from Lake Wigry (north-eastern Poland) and commercially exploited lakes from the southern part of the investigated north-western Poland area. The results of the study of C. albula from northern Poland present a valuable molecular characterization of the populations and can be a starting point for further genetic monitoring.

Aims: A transposition of the long segment of distal ileum in obese Zucker rats improved glucose tolerance 6 months after IT. It was undertaken to compare the gene expression of miRNA-103, -107 and caveolin-1 in the liver of euglycemic groups of IT relative to SHAM operated rats. Main methods: Obese, male Zucker rats underwent either transposition of 50% distal ileum or sham surgery. For determining the gene expression, the Real-Time PCR for caveolin-1 and miRNA-103, -107 was performed. Plasma concentrations of LDL, HDL, TG and total cholesterol were measured with enzymatic colorimetric assays after optimization procedure. Key findings: The Cav-1 expression in liver tissue after ileal transposition was 1.22 times higher compared to the SHAM group (SHAM median 63.58, min 41.3, max 82.4; IT median 77.35, min 60.8, max 95.41, p < 0.001). miRNA-107 expression was significantly downregulated by 0.6-fold in the IT group compared to the SHAM group (SHAM median 507.51, min 236.42, max 721.29; IT median 355.2, min 278.15, max 478.15, p < 0.015. The level of TG was significantly higher after IT surgery (SHAM median 115, min 96, max 143; IT median 153, min 115, max 162, p = 0.001). The total cholesterol plasma levels decreased after IT (SHAM median 178, min 161, max 183; IT median 128, min 103, max 114, p < 0.000001). The LDL plasma level in IT was two-fold lower than in the SHAM (SHAM median 117, min 68, max 151; IT median 58, min 45, max 61, p < 0.000001). Significance: The transposition of 50% of the distal ileum lead to an increase in caveolin-1 and reduction in miR-107 expression compared to those of SHAM group. Endogenous miR-107 is more involved in regulation of the functions of insulin-target liver tissue than miRNA-103. Reduced LDL and cholesterol plasma levels suggest positive effects on lipid metabolism in long-term observations. The present study is the first to show a lack of IT effect regarding triglycerides six months after surgery.

The phosphatidylinositol 3-kinase (PI3K) signaling pathway regulates many important cellular functions. The functional impact of deregulating the PIK3CA gene, encoding the p110α catalytic subunit of PI3K, is validated by frequent gain of function mutations in a range of human cancers. We generated a mouse model with an inducible constitutively active form of PI3K. In this model Cre recombinase activates expression of a myristoylated form of p110α (myr-p110α). The myristoylated version of p110α brings the protein to the cytoplasmic side of the cell membrane, which mimics the normal activation mechanism for the p110α catalytic subunit and activates the PI3K enzyme. Constitutively activated PI3K signaling induced by myr-p110α in all cells of the developing mouse caused lethality during embryonic development. Transgenic Cre;myr-p110α heterozygous embryos displayed morphological malformation and poor vascular development with extremely dilated blood vessels and hemorrhage in the embryo and the extraembryonic yolk sac. Previous studies demonstrated that loss of p110α during embryonic development causes angiogenic disruption and here we show that constitutive activation of p110α by gain of function mutation during development also disrupts vasculogenesis/angiogenesis in what appears to be a similar manner. These finding demonstrate the importance of tight regulation of PI3K signaling during embryonic vasculogenesis/angiogenesis.

Collagen hydrogels were considered as favourable scaffolding for tissue engineering. It was demonstrated that cytokines and siRNAs could be efficiently retained by collagen hydrogels for controlled release thereby enhancing their bioactivities. Basic fibroblast growth factor (bFGF) was a stimulator for osteogenic differentiation of mesenchymal stem cells (MSC), and PPARγ was a key regulator in MSC osteogenic differentiation. However, whether bFGF and PPARγ could play synergetic roles within a 3D matrix to promote MSC osteogenic differentiation was unknown. In the study, bFGF and PPARγ targeting siRNAs were incorporated into collagen hydrogels for MSC cultivation. Their optimal concentrations in collagen hydrogels were determined. The capacity of bFGF/siRNA-carrying hydrogels in supporting osteogenic differentiation of MSCs was systematically evaluated with multimodality of methods, including flow cytometry, quantitative real-time PCR, Western Blotting, as well as ALP activity and calcium content determination. We demonstrated in 3D collagen hydrogel that both bFGF and siRNA molecules were efficiently retained, strengthening their effects on the incorporated MSCs. Osteogenic analysis demonstrated that the in-situ forming hydrogels carrying bFGF and siRNAs potently promoted osteogenic differentiation of incorporated MSCs, significantly superior to pure collagen and bFGF-carrying collagen. Thus, collagen hydrogels functionalized with bFGF and PPARγ targeting siRNAs may be promising in bone tissue engineering.

Leishmaniasis is a disease with a worldwide distribution affecting both humans and animals. There is a need to identify and develop new drugs for the treatment of leishmaniasis. This study showed that crude ethanolic extracts of the red alga Asparagopsis taxiformis have a powerful effect against L. infantum, the prevalent species of the genus Leishmania in the Mediterranean basin. L. infantum demonstrated decreased vitality with increasing concentration of the algal extracts. At a concentration of 40 μg/mL, the extracts achieved 100% mortality of the parasite and the LD 50 value was 25 μg/mL for promastigotes and 9 μg/mL for amastigotes. Algal extracts caused morphological alterations and apoptosis in Leishmania cells. The potential cytotoxic action of crude extracts was investigated by a MTT viability assay on DH82 and Vero cell lines but there was no cytotoxic effect. The potential of red alga A. taxiformis metabolites as anti-leishmanial agents merits further pharmacological investigation.

Wax morphology and chemical composition are widely accepted to be important for the protective properties of the leaf’s surface and also valuable characteristics in plant systematics. The leaves of Sorbus domestica L. and Cotoneaster granatensis Boiss., species of two large genera with intricate taxonomy referred to subtribe Pyrinae, Rosaceae (formerly subfamily Maloideae), were studied by scanning electron microscope (SEM) and performing different methods of wax isolation. The aim of the study was to acquire a suitable, cost and time effective method for wax removal. Chloroform and methanol extractions and freeze-embedding method for direct isolation of the wax crystals were applied. Immersing the leaves for 3 minutes in chloroform was sufficient to extract the waxes whereas the efficiency of the methanol solvent was lower. Wax layers with wellpreserved structures of the crystals from both upper and lower epidermis were successfully transferred to artificial surfaces. The recrystallization experiment demonstrated that waxes from chloroform extracts could recrystallize in in vitro conditions on artificial surfaces. The crystals showed same micromorphology as on the intact leaves. Results of this study could be applied in further analytical researches of the waxes of S. domestica and C. granatensis or other species of the subtribe Pyrinae.

The breeding phenology and reproductive strategies of Porcellio variabilis were investigated, both in field and laboratory, in the population of Béja (North of Tunisia). P. variabilis Lucas, 1946 is characterized by a seasonal reproduction period followed by a sexual rest phase during November, December and January. Mean fecundity showed a great seasonal variation ranging between 29.612 eggs for females caught in the late summer, and 125.920 eggs for those collected in the spring. P. variabilis is an iteroparous species; females were able to produce one (95% of females), two (40% of females) or three broods (20% of females) during their reproduction period. The duration of the gestation period ranged from 34 days in the first brood to 14 days in the third one. Energy allocated to reproduction decreased from the first brood (26.29%) to the last one (4.618%). During the sampling period, eight cohorts were identified, each exhibiting a variable life span estimated around 6 months for cohorts born in early spring, and 12 months for those born in the fall. According to our present results and previous studies in another population of P. variabilis in Tunisia, we discuss factors of variability in breeding phenology in this species.

Gill characteristics and the size and number of erythrocytes were observed for three species of goby: Neogobius melanostomus, N. fluviatilis and N. kessleri. While erythrocyte size was similar in all species, N. fluviatilis had a statistically higher erythrocyte count. Significant differences were found in gill apparatus structure, with N. melanostomus having 25% and 50% greater gill contact area than N. kessleri and N. fluviatilis, respectively. Secretory goblet cells, which produce a protective mucous, were also present in highest numbers in N. melanostomus gill tissues. These physiological and morphological differences may be one reason for differences observed in the invasiveness of these three species.

Two cry1-type genes encoding insecticidal crystal proteins (ICPs) were detected by PCR-RFLP and cloned from Bacillus thuringiensis subsp. galleriae 87. The nucleotide sequences were deposited in GenBank with accession numbers EU679501 and EU679502, and designated as cry1Fb7 and cry1Cb3 respectively by B. thuringiensis Delta- Endotoxin Nomenclature Committee. cry1Cb3 shared 99% homology with other cry1Cb genes. The existence of two additional stop codons indicated cry1Cb3 was a silent gene. The cry1Cb3 was 3531 bp with 38.98% G+C content and its first open reading frame (ORF) encoded a protein of 213 amino acid residues with a calculated molecular weight of 23.8 kDa and a predicted pI value of 4.63. Five amino acid sequence blocks (block 1, block 2, block 3, block 4 and block 5) were found in Cry1Cb3. Translation of cry1Fb7 revealed an ORF of 3525 bp with 39.12% G+C content and a protein with a calculated molecular weight of 133.2 kDa and a predicted pI value of 5.18. Cry1Fb7 had five amino acid sequence blocks (blocks 1, 2, 3, 4 and 5) and three domains (I, II and III), which consisted of 218 residues (Leu 34 to Ala 252 ), 197 residues (Thr 257 to Asp 454 ), and 138 residues (Ile 464 to Glu 600 ), respectively.

Knowledge of protein structure plays a key role in the analysis of protein functions, protein binding, rational drug design, and many other related fields and applications. In this study, a novel feature extraction model based on linear predictive coding (LPC) and position-specific score matrices (PSSM) was proposed to predict structural class from protein sequences. First, the PSI-BLAST tool was employed to transform the original protein sequences into PSSMs. Then, the LPC, a signal processing tool, was applied to extract the features from PSSMs. The selected features were finally fed to a support vector machine to perform the prediction. Cross-validation tests on the four benchmark datasets Z277, Z498, 1189 and 25PDB, showed a significant leap in overall accuracy using the proposed method. Compared to existing methods, our method achieved better performance in prediction of protein structural class.

Biotic homogenization is the increasing similarity of the species composition of communities over time and represents a loss of biodiversity. We analysed changes in weed vegetation over a period of 70 years by comparing three datasets (from 1939, 2002 and 2012) sampled with the same methodology. We present the results of changes in species richness, homogenization and differentiation as expanding neophytes and generalist species. The species richness of weed communities decreased and the number of neophytes in cereal fields and root crops increased over time. The decreased ratio of specialists to generalists in vernal communities and cereal crops indicates homogenization, while the ratio of generalists to specialists increased in root crops.

Introduction: The association of vaspin and visfatin, with a myocardial infarction is still not fully understood. Reduced levels of adenine nucleotides are hallmarks of chronic heart failure. There is little data concerning the relationship between these markers and their changes over time. Material/Methods: The concentration of adenine nucleotides, vaspin and visfatinwere assessed in 41 consecutive patients with acute myocardial infarction one before (day I) and four days after (day IV) percutaneous coronary intervention (PCI) and a control group. Results: Visfatin concentrations were higher before and after PCI vs. control (visfatin I: median 25.55, 20.12 - 30.69 ng/ml; visfatin IV: median 20.79, 16.89 - 25.61 ng/ml vs. control: median 14.94, 10.66 - 25.25 ng/ml; p < 0.0001). Vaspin concentrations were lower before and after PCI vs. control (vaspin I: median 0.18, 0.11 - 0.44 ng/ml; vaspin IV: median 0.24, 0.15 - 0.58 ng/ml vs. control: median 1.303, 1.13 - 2.26 ng/ml, p < 0.00001). Concentrations of visfatin, day I, correlated well to vaspin concentrations (r 2 = 0.201, p = 0.011). ATP levels were significantly lower in patients vs. controls (day I: p = 0.00012; day IV: p = 0.0001). Conclusions: Changes in the analyzed visfatin and vaspin concentrations can be used as potential MI markers. Visfatin serum concentration may be considered a potential marker to differentiate MI over time.

The aim of this work was to investigate the different evolutionary patterns between HBeAg(+) and HBeAg(-) status of genotype B and genotype C of Hepatitis B virus (HBV). Complete HBV genome sequences were obtained from GenBank, and were classified as HBeAg(+) of genotype B, HBeAg(-) of genotype B, HBeAg(+) of genotype C, and HBeAg(-) of genotype C. The results showed that genotype B had lower diversity and a longer phylogenetic tree than genotype C. This means that genotype B was more conserved than genotype C. The genetic distance of HBeAg(+) was shorter than that of HBeAg(-) in genotype B. dN, dS, dN/dS of HBeAg(+) were lower than that of HBeAg(-). In addition, the Core and S genes had lower diversity, and were under lower selective pressure than other genes. We propose that by having different HBeAg HBV receives disparate immune selective pressures, however these different genotypes also receive disparate natural selective pressures. The evolutionary pattern of HBV could be one of the important factors that may affect viral replication in HBV infected patients and could therefore play an important role in the diseases progression.