Bioprospecting of Turbinaria Macroalgae as a Potential Source of Health Protective Compounds

The present study aims to focus on the bioprospecting of marine macroalgae of Turbinaria species, plenteous biomass of the world ocean. Three types of solvents, i.e., H₂O, MeOH/H₂O (80:20, v/v) and hexane/i‐PrOH (50:50, v/v), were used for extraction. Both the biological activity and the pattern of present chemicals were characterized. For the cell proliferation assay, the human embryonic kidney 293 cells, cervix/breast/pancreatic adenocarcinoma, and osteosarcoma cells were used. For the antioxidant activity determination, both intracellular assay with human embryonic kidney and cervix adenocarcinoma cells, as well as the biochemical DPPH test, were employed. To complete the information about macroalgae composition, organic compounds were characterized by the liquid chromatography coupled with high resolution tandem mass spectrometry. Attention was concentrated mainly on the lipidomic profile characterization. In spite the fact that any significant antiproliferative effect was not observed for cancer cells, both the Turbinaria species were shown to be good protectors against the oxidative stress of the non‐cancer cells. Most of the antioxidants were determined in the hexane/i‐PrOH extract. As regards the lipids identified, most of them belonged to the triacylglycerols followed by sphingomyelins, diacylglycerols, and polar (lyso)phospholipids. Additionally to fatty acids with 14, 16 and 18 carbons, also those with odd carbon numbers were frequently present.     

Anthraquinones quinizarin and danthron unwind negatively supercoiled DNA and lengthen linear DNA

The intercalating drugs possess a planar aromatic chromophore unit by which they insert between DNA bases causing the distortion of classical B-DNA form. The planar tricyclic structure of anthraquinones belongs to the group of chromophore units and enables anthraquinones to bind to DNA by intercalating mode. The interactions of simple derivatives of anthraquinone, quinizarin (1,4-dihydroxyanthraquinone) and danthron (1,8-dihydroxyanthraquinone), with negatively supercoiled and linear DNA were investigated using a combination of the electrophoretic methods, fluorescence spectrophotometry and single molecule technique an atomic force microscopy. The detection of the topological change of negatively supercoiled plasmid DNA, unwinding of negatively supercoiled DNA, corresponding to appearance of DNA topoisomers with the low superhelicity and an increase of the contour length of linear DNA in the presence of quinizarin and danthron indicate the binding of both anthraquinones to DNA by intercalating mode.     

Pallenopsis patagonica (Hoek, 1881) – a species complex revealed by morphology and DNA barcoding,with description of a new species of Pallenopsis Wilson, 1881

Pallenopsis patagonica (Hoek, 1881) is one of the most taxonomically problematic and variable pycnogonid species, and is distributed around the southern South American coast, and the Subantarctic and Antarctic areas. We conducted a phylogenetic analysis of mitochondrial cytochrome c oxidase subunit I (COI) sequences of 47 Pallenopsis specimens, including 39 morphologically identified as P. patagonica, five Pallenopsis pilosa (Hoek, 1881), one Pallenopsis macneilli Clark, 1963, one Pallenopsis buphtalmus Pushkin, 1993, and one Pallenopsis latefrontalis Pushkin, 1993. Furthermore, we studied morphological differences between the different COI lineages using light and scanning electron microscopy, including also material from Loman's and Hedgpeth's classical collections, as well as Hoek's type material of P. patagonica from 1881. The molecular results unambiguously reveal that P. patagonica is a complex of several divergent clades, which also includes P. macneilli, P. buphtalmus, and P. latefrontalis. Based on the material available, two major clades could be identified, namely a ‘Falkland’ clade, to which we assign the nominal P. patagonica, and a ‘Chilean’ clade, which is distinct from the ‘Falkland’ clade. We describe the ‘Chilean’ clade as new species, P allenopsis yepayekae sp. nov. Weis, 2013. All molecular results are confirmed by specific morphological characteristics that are discussed in detail and compared with Pallenopsis species closely related to the P. patagonica complex. Our results reveal that P. patagonica is a species‐rich complex that is in need for a thorough taxonomic revision, using both morphological and genetic approaches. © 2014 The Linnean Society of London     

Unequal distribution of genes and chromosomes refers to nuclear diversification in the binucleated Giardia intestinalis

The single-celled parasite Giardia intestinalis (Diplomonadida) has two equally sized nuclei in one cell. The nuclei have been considered identical. We have previously shown that they contain different chromosomal sets and proceed through the cell cycle with some asynchrony. Here, we demonstrate by fluorescence in situ hybridization that several genes from chromosome 5 are lost in one of the two nuclei of the WBc6 Giardia line. The missing segment stretches over at least 50 kb near the 5′ chromosome end. In both WB and WBc6 Giardia cell lines, chromosome 5 is trisomic in one nucleus and monosomic in the other nucleus. The described chromosomal deletion has always been observed at the monosomic chromosome in WBc6; however, the deletion was not detected in the parent line WB. The chromosomal segment was thus initially lost after biological cloning of WB, which gave rise to clone WBc6. We show that Giardia is capable of carrying out gene expression from only one nucleus. The two nuclei display a certain level of diversity, making each of them irreplaceable. The doubled karyomastigonts of diplomonads likely have separate functions both in the mastigont/flagellar organization and in chromosomal and gene content. To our knowledge, our results offer the first methodical approach to differentiating the two, so far indistinguishable nuclei.     

Metabolic control of the membrane fluidity in Bacillus subtilis during cold adaptation

Membrane fluidity adaptation to the low growth temperature in Bacillus subtilis involves two distinct mechanisms: (1) long-term adaptation accomplished by increasing the ratio of anteiso- to iso-branched fatty acids and (2) rapid desaturation of fatty acid chains in existing phospholipids by induction of fatty acid desaturase after cold shock. In this work we studied the effect of medium composition on cold adaptation of membrane fluidity. Bacillus subtilis was cultivated at optimum (40 degrees C) and low (20 degrees C) temperatures in complex medium with glucose or in mineral medium with either glucose or glycerol. Cold adaptation was characterized by fatty acid analysis and by measuring the midpoint of phospholipid phase transition T(m) (differential scanning calorimetry) and membrane fluidity (DPH fluorescence polarization). Cells cultured and measured at 40 degrees C displayed the same membrane fluidity in all three media despite a markedly different fatty acid composition. The T(m) was surprisingly the highest in the case of a culture grown in complex medium. On the contrary, cultivation at 20 degrees C in the complex medium gave rise to the highest membrane fluidity with concomitant decrease of T(m) by 10.5 degrees C. In mineral media at 20 degrees C the corresponding changes of T(m) were almost negligible. After a temperature shift from 40 to 20 degrees C, the cultures from all three media displayed the same adaptive induction of fatty acid desaturase despite their different membrane fluidity values immediately after cold shock.     

Trends in pollen season characteristics of Alnus,Poaceae and Artemisia allergenic taxa in Bratislava,central Europe

Over the period 2002–2019, air temperature and precipitation significantly increased regionally for Bratislava, which could lead to phenological changes in some plant species. This study aimed to analyse the changes in the intensity, timing, and duration of pollen seasons of three allergological important plant taxa (Alnus, Poaceae, Artemisia) in the study area over 18 years. The pollen sampling was performed using a Hirst-type sampler. Mann–Kendall tau test was used to determine trends in pollen season characteristics, while Spearman’s correlation analysis was used to identify the relationships between the characteristics of pollen seasons and both air temperature and precipitation trends. The notable changes in the pollen-season-related features were observed for all analysed taxa. The Alnus pollen season now reaches the peak earlier and its intensity is rising in line with the summer-autumn temperature increasing trend, while unexpectedly intensity and duration of the Artemisia pollen season are declining in line with the increased precipitation and/or temperature trends. On the other hand, the intensity of the Poaceae pollen season is also declining, however, without statistically significant correlations with recorded increases in meteorological parameters considered. This phenomenon is probably related to both the reduction of the area of grasslands due to urbanization and the implementation of effective maintenance of urban green areas (e.g., timely mowing preventing the repeatedly flowering of grasses).

     

Resistin inhibits essential functions of polymorphonuclear leukocytes

The serum levels of resistin, a 12-kDa protein primarily expressed in inflammatory cells in humans, are increased in patients with chronic kidney disease and in those with diabetes mellitus. Both groups of patients have an increased risk of infections mainly as a result of disturbed polymorphonuclear leukocyte (PMNL) functions. Therefore, we investigated the influence of resistin on human PMNLs. Serum resistin concentrations were determined with a sandwich enzyme immunoassay. Using PMNLs from healthy subjects, chemotaxis was tested by the under-agarose method. Flow cytometric assays to measure oxidative burst and phagocytosis were conducted in whole blood. The uptake of deoxyglucose was determined as measure of the PMNL activation state. The activity of intracellular kinases was assessed by Western blotting and by in vitro kinase assays. Resistin inhibited PMNL chemotaxis and decreased the oxidative burst stimulated by Escherichia coli and by PMA, but did not influence PMNL phagocytosis of opsonized E. coli and PMNL glucose uptake. The inhibition of PMNLs by resistin was observed at concentrations found in serum samples of uremic patients, but not in concentrations measured in healthy subjects. Experiments with specific signal transduction inhibitors and measurements of intracellular kinases suggest that PI3K is a major target of resistin. In conclusion, resistin interferes with the chemotactic movement and the stimulation of the oxidative burst of PMNL, and therefore may contribute to the disturbed immune response in patients with increased resistin serum levels such as uremic and diabetic subjects.