Methanol Extracts of 28 Hieracium Species from the Balkan Peninsula – Comparative LC–MS Analysis,Chemosystematic Evaluation of their Flavonoid and Phenolic Acid Profiles and Antioxidant Potentials

Introduction

Hieracium s. str. represents one of the largest and most complex genera of flowering plants. As molecular genetics seems unlikely to disentangle intricate relationships within this reticulate species complex, analysis of flavonoids and phenolic acids, known as good chemosystematic markers, promise to be more reliable. Data about pharmacological activity of Hieracium species are scarce.

Objective

Evaluation of the chemosystematic significance of flavonoids and phenolic acids of methanol extracts of aerial flowering parts of 28 Hieracium species from the Balkans. Additionally, investigation of antioxidant potentials of the extracts.

Methods

Comparative qualitative and quantitative analysis of flavonoids and phenolic acids was performed by LC–MS. Multivariate statistical data analysis included non‐metric multidimensional scaling (nMDS), unweighted pair‐group arithmetic averages (UPGMA) and principal component analysis (PCA). Antioxidant activity was evaluated using three colorimetric tests.

Results

Dominant phenolics in almost all species were luteolin type flavonoids, followed by phenolic acids. Although the investigated Hieracium species share many compounds, the current classification of the genus was supported by nMDS and UPGMA analyses with a good resolution to the group level. Hieracium naegelianum was clearly separated from the other investigated species. Spatial and ecological distances of the samples were likely to influence unexpected differentiation of some groups within H. sect. Pannosa. The vast majority of dominant compounds significantly contributed to differences between taxa. The antioxidant potential of the extracts was satisfactory and in accordance with their phenolics composition.

Conclusions

Comparative LC–MS analysis demonstrated that flavonoids and phenolic acids are good indicators of chemosystematic relationships within Hieracium, particularly between non‐hybrid species and groups from the same location. Copyright © 2017 John Wiley & Sons, Ltd.     

Formation of α- and β-conidia by Phaeocytostroma ambiguum

The formation of conidia in Phaeocytostroma ambiguum on different media and conditions was investigated in this study. Carnation leaf agar (CLA) and a 12 h photoperiod (24/18 °C) provided excellent conditions for the promotion of rapid formation of both alpha (α) and beta (β) conidia in a number of P. ambiguum isolates. The dimensions of α- and β-conidia amounted to 6.0–19.6 × 3.8–7.5 μm and 6.0–24.9 × 1.1–2.6 μm, respectively. They were produced on short or elongate, simple and branched conidiophores. β-conidia have not been described before in P. ambiguum. Intermediate conidia were rarely found. This revised version was published online in August 2006 with corrections to the Cover Date.     

The genusCandida berkhout

The aim of the study was to verify the accuracy of the taxonomic classification of rough variants of the speciesCandida guilliermondii on the basis of comprehensive study of phenetic manifestation and to determine differences in cell wall structure with special reference to polysaccharides (1) According to their phenotype, the test strainsCandida guilliermondii (Cast.) Langeron et Guerra andCandida guilliermondii var.membranaefaciens Lodder et Kreger-Van Rij belong to the speciesCandida guilliermondii, whileCandida guilliermondii var.nitratophila Diddens et Lodder is phenotypically closer toCandida pelliculosa. (2) Observation of native and hydrolysed cell walls in the electron microscope showed no differences between the test strains. (3) The results of X-ray phase analysis of cell walls differentiatedCandida guilliermondii var.nitratophila from the other two, however. (4) Electron microscopy photomicrographs and diffractograms of cell walls indicated that, after 2% HCl extraction at 100 C, the cell walls contain chitin, which is isolated by further extraction with 30% HCl. After 3% NaOH hydrolysis the chitin diffractogram is not clear.     

Antioxidant defence enzyme activities in hepatopancreas, gills and muscle of Spiny cheek crayfish (Orconectes limosus) from the River Danube

The aim of our study was to determine the activity of antioxidant defence (AD) enzymes: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR) and the phase II biotransformation enzyme glutathione-S-transferase (GST) in the hepatopancreas, the gills and muscle of Spiny cheek crayfish (Orconectes limosus) from the River Danube and to compare tissue specificities of investigated enzymes. Our results indicated that both specific and total SOD activities in the hepatopancreas were lower compared to the gills and muscle. Total SOD activity in the gills was lower with respect to that in muscle. CAT and GSH-Px (both specific and total) activities were higher in the hepatopancreas compared to those in the gills and muscle. In the gills the specific and total GR activities were higher than in the hepatopancreas and muscle. The specific and total GST activities were higher in the hepatopancreas compared with the gills and muscle. Our study represents the first comprehensive report of AD enzymes in tissues of O. limosus caught in the River Danube. The noted tissue distributions of the investigated AD enzyme activities most likely reflected different metabolic activities and different responses to environmental conditions in the examined tissues.     

The PECAM-1 gene polymorphism — a genetic marker of myocardial infarction

We investigated a possible association between the C373G (Leu125Val) polymorphism in the platelet endothelial cell adhesion molecule-1 (PECAM-1) and myocardial infarction (MI) among patients with type 2 diabetes (T2DM) in the Slovene population (Caucasians). The study population of this cross-sectional analysis consisted of 452 subjects with T2DM lasting more than 10 years: 142 patients with MI (MI group) and 310 patients (control group) with no history of coronary diseases. There were significant differences of PECAM-1 genotype distribution in patients with MI (CC=28.2%, CG=47.2% and GG=24.6%) compared with subjects in the control group (CC=17.1%, CG=53.5% and GG=29.4%). The multivariate model showed that the CC genotype of the PECAM-1 gene polymorphism (C373G) (OR=1.9, 95% CI 1.2–3.0, P=0.007) was an independent risk factor for MI. The C allele frequency was also significantly higher (P=0.005) in MI (51.8%) than in control subjects (41%). In addition, our study revealed the connection between smoking habits, the duration of diabetes and the total and LDL cholesterol serum levels and MI in Slovene T2DM patients. We suggest that the tested polymorphism of PECAM-1 (C373G) is associated with MI. Therefore, it might be used as genetic marker of MI in T2DM.     

Molecular typing and antimicrobial resistance of <Emphasis Type="Italic">Salmonella</Emphasis> Enteritidis isolated from poultry,food, and humans in Serbia

Molecular typing and resistotyping coupled with gyrA single nucleotide polymorphism (SNP) of 60 Salmonella Enteritidis (SE) isolates originated from poultry, food, and humans in Serbia is described. Molecular fingerprinting was performed by randomly amplified polymorphic DNA (RAPD) using four primers, and the diversity index (D) was 0.688. In combination with resistotyping and gyrA SNP, D increased to 0.828. A total of 23 genetic groups were obtained. When four RAPD primers were combined, epidemic isolates from a fast-food restaurant outbreak were clustered in a distinctive genetic group. Among 60 SE strains, three had multiple resistances to three or more antibiotics. Nine strains were resistant to nalidixic acid (NAL; a non-fluorinated quinolone). The mutations in quinolone resistance-determining region (QRDR) found in NAL-resistant strains were attributed to Asp⁸⁷ → Asn in six strains, Asp⁸⁷ → Gly in one strain, and Ser⁸³ → Phe in one strain. One NAL-resistant strain had no mutations in QRDR, suggesting another mechanism of resistance.     

Characterization of velogenic Newcastle disease viruses isolated from dead wild birds in Serbia during 2007

Avian paramyxoviruses type 1 or Newcastle disease viruses (NDV) are frequently recovered from wild birds and such isolates are most frequently of low virulence. Velogenic NDV are usually recovered from poultry and only occasionally from wild birds. Five NDV isolates were obtained from carcasses of four wild bird species during 2007 in Serbia: Mallard (Anas platyrhynchos), Eurasian Sparrowhawk (Accipiter nisus), feral Rock Pigeon (Columba livia), and Eurasian Collared Dove (Streptopelia decaocto). All the isolates have a typical fusion protein cleavage site motif of velogenic viruses ((112)R-R-Q-K-R-F(117)). The highest homology (99%) for the nucleotide sequences spanning the M and F gene of the studied isolates was with the genotype VII NDV isolate Muscovy duck/China(Fujian)/FP1/02. Phylogenetic analysis based on a partial F gene sequence showed that the isolates from wild birds cluster together with concurrent isolates from poultry in Serbia within the subgenotype VIId, which is the predominant pathogen involved currently in Newcastle disease outbreaks in poultry worldwide. It is unlikely that the wild birds played an important role in primary introduction or consequent spread of the velogenic NDV to domestic poultry in Serbia, and they probably contracted the virus from locally infected poultry.     

Effects of Dietary Form of Selenium on Its Distribution in Eggs

The objective of this experiment was to investigate the selenium distribution in eggs from hens fed diets supplemented with Se from sodium selenite (SS) or selenium-enriched yeast (SY). One-day-old female chickens of Hy-Line Brown breed were randomly divided into four groups according to dietary treatments and, for the subsequent 9?months, were fed diets which differed only in the form or amount of Se supplemented. During the whole experiment, group 1 (control) was fed basal diet (BD) with only background Se level of 0.13?mg/kg dry matter (DM). Diets for groups 2 and 3 consisted of BD supplemented with an Se dose of 0.4?mg/kg DM either in the form of SS or SY, respectively. Group 4 was fed BD supplemented with 0.9?mg Se/kg DM from SY. After 9?months of dietary treatments, the Se levels in egg yolk and albumen from hens fed unsupplemented diet were almost identical whereas eggs from hens given diet supplemented with SS showed significantly higher Se deposition in yolk than in albumen (P?相似文献   

Composition, antimicrobial and antioxidant activity of the extracts of Eryngium palmatum Pančić and Vis. (Apiaceae)

The chemical composition, antimicrobial and antioxidant activity of Eryngium palmatum, an endemic plant species from the Balkan Peninsula, were investigated. The flavonoids apigenin (9.5±0.3 mg g^?1) and apigenin 7-O-glucoside (2.4±0.1 mg g^?1) were determined in a methanol extract of aerial parts using HPLC analysis. The methanol extract of roots contained catechin (5.0±0.1 mg g^?1), epicatechin (2.9±0.1 mg g^?1), chlorogenic acid (1.6±0.0 mg g^?1), gallic acid (0.9±0.0 mg g^?1) and rosmarinic acid (0.9±0.2 mg g^?1). GC-FID and GCMS analysis of a chloroform extract of aerial parts showed that the main volatile constituents were falcarinol, linoleic acid, hexadecanoic acid and methyl linoleate (comprising 32.6%; 24.4%; 19.9; 13.2% of the volatile fraction, respectively), while octanoic acid, tetradecanol and dodecanol dominated in the chloroform extract of the roots (34.9%; 25.8%; 22.2% of the volatile fraction, respectively). Investigation of antimicrobial activity by broth microdilution showed that the methanol and chloroform extracts of aerial parts and roots exerted a significant effect (MIC 3.5–15.6 μg mL^?1) against tested Gram-positive and Gram-negative bacteria. The methanol extracts of aerial parts or roots exerted moderate ferric reducing antioxidant power, DPPH radical scavenging activity and hydroxyl radical scavenging activity.