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International audience; Background: Rasagiline, a monoamine oxidase B inhibitor with neuroprotective potential in Parkinson's disease, has shown a disease-modifying effect in the SOD1-Gly93Ala low-expressing mouse model of amyotrophic lateral sclerosis, both alone and in combination with riluzole. We sought to test whether or not rasagiline 1 mg/day can prolong survival in patients with amyotrophic lateral sclerosis also receiving riluzole.Methods: Patients with possible, probable, or definite amyotrophic lateral sclerosis were enrolled to our randomised, placebo-controlled, parallel-group, double-blind, phase 2 trial from 15 German network for motor neuron diseases (MND-NET) centres (university hospitals or clinics). Eligible patients were aged at least 18 years, had onset of progressive weakness within the 36 months before the study, had disease duration of more than 6 months and less than 3 years, and had a best-sitting slow vital capacity of at least 50%. After a 4-week screening period, eligible patients were randomly assigned (1:1) to receive either rasagiline (1 mg/day) or placebo in addition to riluzole (100 mg/day), after stratification for site of onset (bulbar or spinal) and study centre. Patients and all personnel assessing outcome parameters were masked to treatment allocation. Patients were followed up 2, 6, 12, and 18 months after randomisation. The primary endpoint was survival time, defined as the time to death or time to study cutoff date (ie, the last patient's last visit plus 14 days). Analyses of primary outcome and safety measures were done in all patients who received at least one dose of trial treatment (intention-to-treat population). The trial is registered with ClinicalTrials.gov, number NCT01879241.Findings: Between July 2, 2013, and Nov 11, 2014, 273 patients were screened for eligibility, and 252 patients were randomly assigned to receive rasagiline (n=127) or placebo (n=125). 126 patients taking rasagiline and 125 taking placebo were included in the intention-to-treat analysis. For the primary outcome, the survival probability at the end of the study was 0·43 (95% CI 0·25-0·59) in the rasagiline group (n=126) and 0·53 (0·43-0·62) in the placebo group (n=125). The estimated effect size (hazard ratio) was 0·91 (one-sided 97·5% CI -infinity to 1·34; p=0·31). Rasagiline was well tolerated, and most adverse events were due to amyotrophic lateral sclerosis disease progression rather than treatment; the most frequent of these were dysphagia (32 [25%] taking rasagiline vs 24 [19%] taking placebo) and respiratory failure (25 [20%] vs 31 [25%]). Frequency of adverse events were comparable between both groups.Interpretation: Rasagiline was safe in patients with amyotrophic lateral sclerosis. There was no difference between groups in the primary outcome of survival, although post-hoc analysis suggested that rasagiline might modify disease progression in patients with an initial slope of Amyotrophic Lateral Sclerosis Functional Rating Scale Revised greater than 0·5 points per month at baseline. This should be confirmed in another clinical trial.Funding: Teva Pharmaceutical Industries.

Caffeine and selective PDE inhibitors are widely used in clinical management of preterm and term neonates. However, little is known about how these compounds interact with the neonatal adaptive immune system. We aimed to describe the effects of caffeine, milrinone and sildenafil on the activation and cytokine production of T cells from umbilical cord blood (UCB) compared to adult peripheral blood (APB). We isolated mononuclear cells from 10 APB and 6 UCB samples. We assessed intracellular cytokine production (IFN-γ, IL-2, IL-4, IL-6, IL-17) of stimulated CD4 cells and parameters of calcium influx and ROS production following treatment with caffeine, milrinone, sildenafil, dbcAMP or a specific A2A receptor antagonist, ZM241385 using flow cytometry. In ABP, only ZM241385 caused a 1.14-fold increase in calcium influx, while all compounds increased calcium influx in UCB. This effect was more pronounced in case of caffeine (1.41-fold) and dbcAMP (1.3-fold) compared to milrinone (1.22-fold), sildenafil (1.23-fold) or ZM241385 (1.23-fold). Intracellular levels of the studied cytokines were unaffected by the applied compounds in both APB and UCB samples. Caffeine increases calcium influx upon activation in neonatal T lymphocytes to a larger extent than milrinone or sildenafil. This effect appears to be mediated primarily via increased cAMP levels rather than A2A receptor inhibition. Overall, the application of caffeine, sildenafil or milrinone does not appear to have immunosuppressive effects on neonatal T cells.

This study summarizes the results of radiocarbon dating and age-depth modelling in four mountain lakes (Brazi, Gales, Bucura and Lia) in the Retezat Mountains. Altogether 69 AMS 14 C measurements were performed on these deposits, mostly on terrestrial plant macrofossils. In several cases, plant macrofossils, aquatic animal remains (Cladoceran eggs) and bulk sediment were measured from the same depth allowing for the comparison of various sediment components in terms of their dating potential. Age-depth models were developed using both Bayesian modelling with the BACON and smooth spline curve fitting with the CLAM software. In this study Bayesian models are applied to identify outlier 14 C dates, while smooth spline models are used to model sediment accumulation age-depth relations, as these are more likely to follow natural deposition time changes of lake sediments. Changes in sediment deposition times (DT) in relation to catchment size and climate are also studied. DT varied considerably in the late glacial (LG) part of the records. The sediment sequence characterised by high deposition rates during the late glacial (DT maximum around 100–110 years cm −1 ) was defined by small catchment size (Lake Brazi, 6 ha; surface area: 0.5 ha). In contrast, much slower LG sediment deposition in the southern slope lake characterised by large catchment area (Lake Lia, 171 ha, 20 years cm −1 ), principally reflecting strong erosion in these catchment areas at times when vegetation cover was scarce. Holocene was characterised again by variable DT values, but only Lake Gales showed extreme values: 62–110 years between 10,900 and 6050 cal yr BP, followed by much slower sediment deposition in the last 6000 years (av. 29 years cm −1 ). Generally, sediment deposition times were largely dependent on the stability and vegetation cover of the slope in the case of high altitude deep lakes, while lower altitude, shallow lakes showed slower sediment deposition time in the early and mid-Holocene, when summer insolation was higher than today. This reflects that in these shallow lakes in-lake organic production probably increased with summer insolation, which was a significant driver in the rate of deposition.

How genetic diversity of populations reacts to neutral or adaptive processes such as population bottlenecks, immigration or local adaptation are central questions of population genetics. They may be directly answered through ancient DNA analysis, however such studies in plants are remarkably scarce, owing to the difficulty of gaining population-scale samples with good DNA preservation. Plant macrofossils are abundant in lake sediments, and here we assessed if they can be valuable material for population genetic studies. We extracted ancient DNA from early to late Holocene seed and needle remains of Norway spruce (Picea abies L. Karst) recovered from the sediment of a lake in the Retezat Mts. (South Carpathians). We aimed to reveal whether deforestations starting in the late Holocene and reaching maximum in the 19th and early 20th centuries have caused decreased genetic diversity in the modern Norway spruce population of the Retezat Mts. In a combined approach, we sequenced two plastid regions and generated detailed paleobotanical data to reconstruct the population history of Norway spruce. Our results show that genotyping success of the ancient material was high in samples younger than c. 2000-year-old, but was very low in early Holocene samples, and needles allowed genotyping with higher success compared to seeds. We discovered macrofossil samples that contained DNA from more than a single Norway spruce individual. This phenomenon, together with the possible multiple sampling of the same individual needs to be kept in mind when interpreting the genetic data obtained from macrofossils. Our results overall indicate a genetic diversity decrease during the late Holocene. We demonstrate that macrofossils preserved in sediments are invaluable material for population genetic studies.

This paper focuses on the Late Glacial (LG) and Holocene regional and local establishment times of several tree and shrub species in the Retezat Mountains (South Carpathians) using pollen, stomata and plant macrofossil records from four lakes situated at different altitudes. We used the empirical and rational pollen percentage limits to infer the local presence and population expansion/recession times. Our results show that Pinus mugo and P. sylvestris spread first at 15,100 cal yr BP in the mountain. P. mugo arrived at high altitudes (1740 m) already at 13,600 cal yr BP. The expansion of Larix decidua was weakly recorded in the pollen diagrams, but the plant macrofossil and stomata records suggested its Late Glacial expansion mainly on the northern slope and local presence since 14,900 cal yr BP at 1740 m. The species likely went extinct in the Late Holocene (around 3330 cal yr BP). The expansion of Picea abies and Ulmus commenced earlier on the southern slope, from 15,200 to 14,400 cal yr BP respectively, compared to the northern slope, where these species expanded from 14,300 to 13,850 cal yr BP. The following step was the expansion of Quercus at 13,900 cal yr BP. Following the Younger Dryas decrease of temperate deciduous taxa and P. abies, their expansion on the northern flank was in the order of P. abies (11,750), Ulmus (11,600), Quercus (11,100), Corylus (10,600), Carpinus betulus (7100), Abies alba (5100) and Fagus sylvatica (5000 cal yr BP). Timing of the Early Holocene tree expansions on the southern flank is ambiguous due to uncertainties in the Early Holocene age-depth model of Lake Lia; however, the mid-Holocene expansion of C. betulus and A. alba were well-dated in both southern slope pollen records to 7700 and 6100 cal yr BP, respectively, and we inferred several hundred years earlier expansion on the southern flank that can be explained by climatic differences. Higher solar radiation results in longer growing seasons and milder winters on the southern flank, which likely facilitated the earlier spread of these trees. For several woody taxa studied in this paper, last glacial maximum refugial populations can be inferred either locally in the mountain (P. abies, P. mugo, P. sylvestris, L. decidua) or in the region (Quercus, Ulmus, C. betulus, F. sylvatica) on the basis of their pollen presence since at least 15,700–16,000 cal yr BP and early increase during the LG. We compared the pollen inferred population expansion times of several tree taxa along a south-north transect in the Romanian Carpathians, and found that the Early Holocene high altitude expansion of A. alba (10,650–9400 cal yr BP) is exceptional in the region. We also demonstrated that the Holocene mass expansion of Quercus and C. avellana started earliest in the Retezat, and at least two centuries later in other sites in the Romanian Carpathians. We also demonstrated that certain tree species had preference for survival and/or early expansion on the northern slopes (A. alba, L. decidua, F. sylvatica), while others on the southern slopes (C. avellana, C. betulus).

Pluripotent stem cell derived human neuronal progenitor cells (hPSC-NPCs) and their mature neuronal cell culture derivatives may efficiently be used for central nervous system (CNS) drug screening, including the investigation of ligand-induced calcium signalization. We have established hippocampal NPC cultures derived from human induced PSCs, which were previously generated by non-integrating Sendai virus reprogramming. Using established protocols these NPCs were differentiated into hippocampal dentate gyrus neurons. In order to study calcium signaling without the need of dye loading, we have stably expressed an advanced calcium indicator protein (GCaMP6fast) in the NPCs using the Sleeping Beauty transposon system. We observed no significant effects of the long-term GCaMP6 expression on NPC morphology, gene expression pattern or neural differentiation capacity. In order to compare the functional properties of GCaMP6-expressing neural cells and the corresponding parental cells loaded with calcium indicator dye Fluo-4, a detailed characterization of calcium signals was performed. We found that the calcium signals induced by ATP, glutamate, LPA, or proteases - were similar in these two systems. Moreover, the presence of the calcium indicator protein allowed for a sensitive, repeatable detection of changes in calcium signaling during the process of neurogenesis and neuronal maturation.

The commercial enzymatic preparation, Pectinex Ultra SP-L, was successfully immobilized on activated chitosan particles for production of lactulose-based galacto-oligosaccharide (LuOS). The main degrees of polymerization (DP) of LuOS produced in this bioreactor were DP3 and DP4. The immobilization yield in term of expressed activity and recovery activity were 71% and 36.05%, respectively. The optimum temperature (60° C) was not affected by immobilization process, while the optimum pH was shifted from pH 5.5 to pH 4.5. In addition, a thermal stability of the immobilized enzyme was better than that of free enzyme. The half-life time of hydrolytic activity increased from 2.5 days (free enzyme) to 14 days (immobilized enzyme). Hydrolytic KM values were 651.3 mM and 187.9 mM lactulose for free enzyme and immobilized enzyme, respectively. Furthermore, the immobilized enzyme showed high reusability. The yield of LuOS formation was more than 60% after 33 cycles of reuses. In the case of trans-galactosylation, a model with 8 reaction rates was build using five-step reaction pathways with different initial lactulose concentrations. This catalytic model was validated well for a long-term period (27 days). These results strongly support to develop a continuous bioconversion to synthesize functional food ingredients, prebiotic LuOS.

Pomalidomide (POM), a potent anticancer thalidomide analogue was characterized in terms of cyclodextrin complexation to improve its aqueous solubility and maintain its anti-angiogenic activity. The most promising cyclodextrin derivatives were selected by phase-solubility studies. From the investigated nine cyclodextrins – differing in cavity size, nature of substituents, degree of substitution and charge – the highest solubility increase was observed with sulfobutylether-β-cyclodextrin (SBE-β-CD). The inclusion complexation between POM and SBE-β-CD was further characterized with a wide variety of state-of-the-art analytical techniques, such as nuclear magnetic resonance spectroscopy (NMR), infrared spectroscopy (IR), circular dichroism spectroscopy, fluorescence spectroscopy as well as X-ray powder diffraction method (XRD). Job plot titration by NMR and the AL-type phase-solubility diagram indicated 1:1 stoichiometry in a liquid state. Complementary analytical methods were employed for the determination of the stability constant of the complex; the advantages and disadvantages of the different approaches are also discussed. Inclusion complex formation was also assessed by molecular modelling study. Solid state complexation in a 1:1 M ratio was carried out by lyophilization and investigated by IR and XRD. The complex exhibited fast-dissolution with immediate release of POM, when compared to the pure drug at acidic and neutral pH. Kinetic analysis of POM release from lyophilized complex shows that Korsmeyer-Peppas and Weibull model described the best the dissolution kinetics. The cytotoxicity of the complex was tested against the LP-1 human myeloma cell line which revealed that supramolecular interactions did not significantly affect the anti-cancer activity of the drug. Overall, our results suggest that the inclusion complexation of POM with SBE-β-CD could be a promising approach for developing more effective POM formulations with increased solubility., Pomalidomide, Cyclodextrin complexation, Inclusion complex, Solubility, Comparative dissolution, Pomalyst®

Aim Although Crohn’s disease (CD) is an extensively investigated autoimmune condition, knowledge on early phase activation of lymphocytes, especially CD8+ Tc cells is scarce. Our aim was to investigate the calcium influx characteristics of CD8+ cells upon activation as well as the expression and function of Kv1.3 and IKCa1 lymphocyte potassium channels. Methods We took peripheral blood from 12 healthy controls, 23 CD children on conventional therapy and 6 severe CD children before and after infliximab therapy. Intracellular calcium levels were monitored in CD8+ lymphocytes using flow cytometry. Results In CD treated with standard therapy calcium response during activation was elevated. This was not affected by the inhibition of Kv1.3 or IKCa1 potassium channels. After the switch to infliximab potassium channel function and expression of CD8+ lymphocytes were comparable to healthy controls in severe CD. Conclusion Calcium handling of CD8+ lymphocytes is altered in pediatric CD, which is normalized by infliximab therapy.

Plants belonging to the genus Rumex (family Polygonaceae) are used worldwide in traditional medicine for the treatment of various diseases caused by different microorganisms (e.g. bacteria-related dermatologic conditions, dysentery and enteritis). The present study focused on the antibacterial screening of Rumex species native to the Carpathian Basin, and isolation of compounds from one of the most efficient species, Rumex aquaticus. The antibacterial effects of n-hexane, chloroform and aqueous fractions of methanol extracts prepared from different parts of 14 Rumex species (R. acetosella, R. acetosa, R. alpinus, R. aquaticus, R. conglomeratus, R. crispus, R. hydrolapathum, R. obtusifolius subsp. obtusifolius, R. obtusifolius subsp. subalpinus, R. patientia, R. pulcher, R. scutatus, R. stenophyllus and R. thyrsiflorus) were investigated against Staphylococcus epidermidis, S. aureus, MRSA, Bacillus subtilis, Moraxella catarrhalis, Streptococcus pyogenes, S. pneumoniae, S. agalactiae, Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae using the disc diffusion method. Mainly the n-hexane and chloroform extracts prepared from the roots of the plants displayed high antibacterial activity (inhibition zones > 15 mm) against one or more bacterial strains. The highly active extracts of the aerial part and root of R. aquaticus were subjected to a multistep separation procedure. 19 Compounds, among them naphthalenes (musizin, and its glucoside, torachrysone-glucoside, 2-methoxystypandrone), anthraquinones (emodin, chrysophanol, physcion, citreorosein, chrysophanol-8-O-glucoside), flavonoids (quercetin, quercetin-3,3′-dimethylether, isokaempferide, quercetin 3-O-arabinoside, quercetin 3-O-galactoside, catechin), stilbenes (resveratrol, piceid), and 1-stearoylglycerol were isolated from the plant. The antibacterial activities of isolated compounds were determined, and it was observed that especially naphthalenes exerted remarkable antibacterial effects against several bacterial strains.

There are numerous applications of quantitative PCR for both diagnostic and basic research. As in many other techniques the basis of quantification is that comparisons are made between different (unknown and known or reference) specimens of the same entity. When the aim is to compare real quantities of different species in samples, one cannot escape their separate precise absolute quantification. We have established a simple and reliable method for this purpose (Ct shift method) which combines the absolute and the relative approach. It requires a plasmid standard containing both sequences of amplicons to be compared (e.g. the target of interest and the endogenous control). It can serve as a reference sample with equal copies of templates for both targets. Using the ΔΔCt formula we can quantify the exact ratio of the two templates in each unknown sample. The Ct shift method has been successfully applied for transposon gene copy measurements, as well as for comparison of different mRNAs in cDNA samples. This study provides the proof of concept and introduces some potential applications of the method; the absolute nature of results even without the need for real reference samples can contribute to the universality of the method and comparability of different studies.

Flow cytometry is a common approach to study invertebrate immune cells including earthworm coelomocytes. However, the link between light-scatter- and microscopy-based phenotyping remains obscured. Here we show, by means of light scatter-based cell sorting, both subpopulations (amoebocytes and eleocytes) can be physically isolated with good sort efficiency and purity confirmed by downstream morphological and cytochemical applications. Immunocytochemical analysis using anti-EFCC monoclonal antibodies combined with phalloidin staining has revealed antigenically distinct, sorted subsets. Screening of lectin binding capacity indicated wheat germ agglutinin (WGA) as the strongest reactor to amoebocytes. This is further evidenced by WGA inhibition assays that suggest high abundance of N-acetyl-d-glucosamine in amoebocytes. Post-sort phagocytosis assays confirmed the functional differences between amoebocytes and eleocytes, with the former being in favor of bacterial engulfment. This study has proved successful in linking flow cytometry and microscopy analysis and provides further experimental evidence of phenotypic and functional heterogeneity in earthworm coelomocyte subsets.

We show a new approach to evaluate eight different Ocimum basilicum L. gene bank accessions. Based on the characteristics of varieties, we compared gene bank accessions with the sum of ranking differences (SRD) method, which sets priorities among the varieties quickly and efficiently. The results are shown with a suitable graphical evaluation too. SRD running on the original data results one SRD value of each varieties. We applied leave-one-out (LOO) cross-validation to characterize uncertainty of the SRD values, after that we run a re-evaluation of SRD on these data. As a ranked true location of gene bank accession either can only be defined by statistical tests therefore Wilcoxon matched pairs test and Sign test were used to define the pairwise comparison in rankings of LOO SRDnorm values. ’M. Grunes’ variety was the best performing of the selected gene bank stored accessions on the given environmental conditions.

In neuroscience the use of a microelectrode array allows the detection of neuroelectric signals with high temporal resolution in a confined space within the tissue, while two-photon laser scanning microscopy provides high spatial resolution in a wide region of interest. The combination of these two techniques promises better understanding of the operation of neural pathways. To facilitate this connection, we studied the direct electrochemical deposition of the conductive polymer poly-2,3-ethylenedioxy-thiophene onto different Pt and Pt/Ir electrode surfaces from non-aqueous solvents, such as ionic liquid and propylene carbonate. We show the effects of electrochemical deposition technique (pulsed or continuous), monomer concentration range and solvent electrolyte type on the formation of photoluminescent - conductive films. For these variables we determined the optimal deposition parameters given as 0.025–0.050 M EDOT monomer concentration in BMIMBF4 ionic liquid and the use of pulsed deposition process to form an adherent, uniform functional electrode coating.

ABCG2 is a multidrug transporter with wide substrate specificity, and is believed to protect several cell types from various xenobiotics and endobiotics. This "guardian" function is important in numerous cell types and tissue barriers but becomes disadvantageous by being responsible for the multidrug resistance phenotype in certain tumor cells. ABCG2 regulation at the protein level has already been extensively studied, however, regulation at the mRNA level, especially the functional role of the various 5' untranslated exon variants (5' UTRs) has been elusive. In the present work, we describe a comprehensive characterization of four ABCG2 mRNA variants with different exon 1 sequences, investigate drug inducibility, stem cell specificity, mRNA stability, and translation efficiency. Although certain variants (E1B and E1C) are considered as "constitutive" mRNA isoforms, we show that chemotoxic drugs significantly alter the expression pattern of distinct ABCG2 mRNA isoforms. When examining human embryonic stem cell lines, we provide evidence that variant E1A has an expression pattern coupled to undifferentiated stem cell stage, as its transcript level is regulated parallel to mRNAs of Oct4 and Nanog pluripotency marker genes. When characterizing the four exon 1 variants we found no significant differences in terms of mRNA stabilities and half-lives of the isoforms. In contrast, variant E1U showed markedly lower translation efficiency both at the total protein level or regarding the functional presence in the plasma membrane. Taken together, these results indicate that the different 5' UTR variants play an important role in cell type specific regulation and fine tuning of ABCG2 expression.

The emerging resistance of tumor cells against methotrexate (MTX) is one of the major limitations of the MTX treatment of tumorous diseases. The disturbance in the polyglutamation which is a main step in the mechanism of methotrexate action is often the reason of the resistance. Delivery of polyglutamylated MTX into cells may evade the mechanisms that are responsible for drug resistance. In this study conjugates of methotrexate and its pentaglutamylated derivatives with cell-penetrating peptides - penetratin and octaarginine - were investigated. The cellular-uptake and in vitro cytostatic activity of conjugates were examined on breast cancer cell cultures (MDA-MB-231 as resistant and MCF-7 as sensitive cell culture). These cell cultures showed very different behaviour towards the conjugates. Although the presence of pentaglutamyl moiety significantly decreased the internalisation of conjugates, some of them were significantly active in vitro. All of the conjugates were able to penetrate in some extent into both cell types, but only the conjugates of penetratin showed in vitro cytostatic activity. The most effective conjugates were the MTX-Glu5-Penetratin(desMet) and MTX-Glu5-GFLG-Penetratin(desMet). The latter was effective on both cell cultures while the former was active only on the resistant tumor cells. Our results suggest that the translocation of polyglutamylated MTX may be a new way to treat sensitive and more importantly resistant tumors. While both penetratin and octaarginine peptides were successfully used to deliver several kinds of cargos earlier in our case the activity of penetratin conjugates was more pronounced.

Ketogenic diet is a dietetic approach for the treatment of refractory childhood epilepsy. In this experiment we aimed to assess the total lipid content enrichment capability of different however, less studied fats, as well as the possibilities to transfer unsaturated fatty acids and antioxidants into different foods, as these substances proved to be relevant in the mechanism of the ketogenic diet. We also determined the stability of different fats during food preparation and short-term storage. French fries, meatloaf and fried zucchini were prepared with sunflower oil, olive oil, coconut oil, rape oil and lard. The total lipid contents, iodine values, rancidities and antioxidant-activities of the samples were ascertained immediately after the preparation and after 3-day-storage on 4 °C. Our results indicate that the different foods are distinct in their lipid uptake capabilities, while the fats cannot be distinguished by their penetrance abilities into the foods. Sunflower and rape oils proved to be the best for unsaturated fatty acid enrichment but as rape oil proved to be sensitive to both heat and storage time, sunflower oil seems to be the best option. Anitoxidant-acitvity did not show any tendencies.

Cholesteryl lipoprotein semisynthesis was accomplished via a copper-catalyzed azide–alkyne cycloaddition on the surface of live cells. In this convergent synthesis an azido-cholesterol was introduced into the cell membrane without the application of detergents followed by conjugation of the C-terminal alkyne modified protein. This cytocompatible method resulted in a folded membrane-anchored protein containing a small molecule fluorophore in the lipid headgroup.

The present research focused on the preparation of TiO2/WO3 (76/24 wt. %) composite photocatalysts for the photodegradation of methyl orange under UV light. The composites were prepared by the adjustment of the surface charge of the individual semiconductors, achieving a homogeneous distribution of the components within the composite. This resulted in a homogeneous distribution of the components within the composite. The individual WO3 and TiO2 nanocrystals were obtained either by sol–gel or by hydrothermal synthesis. The produced nanomaterials were characterized by using TEM, SEM, XRD and DRS. Major differences in their photocatalytic performance depended on the nature of the precursors and the synthesis pathway.

A 45.8 m high tower is designed. The limitation of the horizontal displacement of the tower top is the main active design constraint. The tower consists of two main parts. The upper 23.8 m high part is designed as a truss structure welded from square hollow section (SHS) rods. The loads are as follows: dead mass of the conductors, wind, ice and break of conductors. The combination of wind and ice loads is calculated according to the new standard EN 50341-1: 2012. The lower 22 m high part is constructed from a circular cylindrical shell with a limited diameter of 2200 mm. To satisfy the displacement constraint, a simple shell should have a thickness of 41 mm, which is unsuitable for forming and welding. The applied new structural version of a cellular shell consists of two concentric shells and longitudinal stiffeners welded between them. This cellular shell has thicknesses of 24 mm, which is much more suitable for fabrication

Ethnopharmacological relevance The approximately 200 species of the genus Rumex (sorrel, Polygonaceae) are distributed worldwide (European, Asian, African and American countries). Some species have been used traditionally as vegetables and for their medicinal properties. Based on the traditional knowledge, different phytochemical and pharmacological activities have been at the focus of research. This review aims to provide an overview of the current state of knowledge of local and traditional medical uses, chemical constituents, pharmacological activities, toxicity, and safety of Rumex species, in order to identify the therapeutic potential of Rumex species and further directions of research. Materials and methods The selection of relevant data was made through a search using the keyword “Rumex” in “Scopus”, “Google Scholar”, “Web of Science”, “PubMed”, and “ScienceDirect” databases. Plant taxonomy was validated by the databases “The Plant List”, and “ Mansfeld's Encyclopedia ”. Additional information on traditional use and botany was obtained from published books and MSc dissertations. Results This review discusses the current knowledge of the chemistry, the in vitro and in vivo pharmacological studies carried out on the extracts, and the main active constituents, isolated from plants of genus Rumex . Although, there are about 200 species in this genus, most of the phytochemical and pharmacological studies were performed on up to 50 species. The aerial parts, leaves and roots of the plants are used as vegetables and for the treatment of several health disorders such as mild diabetes, constipation, infections, diarrhoea, oedema, jaundice, and as an antihypertensive, diuretic and analgesic and in case of skin, liver and gallbladder disorders, and inflammation. Many phytochemical investigations on this genus confirmed that Rumex species are rich in anthraquinones, naphthalenes, flavonoids, stilbenoids, triterpenes, carotenoids, and phenolic acids. Moreover, it draws the attention that high level of oxalic acid in some species can cause toxicity (kidney stones) if consumed large quantity. Conclusions This review confirms that some Rumex species have emerged as a good source of the traditional medicine for treatment of inflammation, cancer and different bacterial infections and provides new insights for further promising investigations on isolated compounds, especially quercetin 3- O -glucoside, emodin, nepodin, torachrysone, and trans-resveratrol to find novel therapeutics and aid drug discovery. In addition, hepatoprotective, antiviral and antidiabetic activities should have priority in future pharmacological studies. However, for applying species to prevent or treat various diseases, additional pharmacological studies are needed to find the mechanism of actions, safety and efficacy of them before starting clinical trials.

In our former study, we proved that sperm injected into the ovary by catheter through the oviduct 10-12 h before hormonally induced ovulation, retains its fertilizing capacity for several hours. Using this method, we produced viable larvae via in vitro fertilisation. The aim of this study was to investigate the time-dependent fertilizing capacity of sperm which were introduced into ovary. Sperm samples (2 mL sperm / bodyweight kg) were incubated in gonad lobes 5, 10, 15, 20, 25, 36 and 48 h before the gamete stripping. Ovulation was induced by extracted carp pituitary (CPE) hormonal administration (5 mg CPE / BW kg). There were no statistical differences (p