Genetic patterns in range-edge populations of Vaccinium species from the central Balkans: implications on conservation prospects and sustainable usage

Vaccinium myrtillus L., Vaccinium uliginosum L. and Vaccinium vitis-idaea L. are perennial, cold-adapted clonal shrubs distributed throughout Europe, northern Asia and North America. Due to their usage in food (berries) and pharmaceutical industry (berries and leaves), their natural populations are exposed to anthropogenic and other impacts that affect their genetic make-up. We analyzed 14 fragmentary distributed and small-sized peripheral populations of these species from the Balkans, which represents the southeastern-European marginal area of their wide European distributions, using RAPD molecular markers. The contemporary genetic patterns in all three species within the Balkans were generally similar, and in comparison to previous reports on populations of these species found in northward Europe, where they have a more continuous distribution, the levels of genetic diversity were more or less halved, genetic differentiation was several times higher, gene flow exceptionally low, and the expected prevalence of clonal individuals was lacking. The population dynamics of all three species within the Balkans was complex and distinct, and was characterized by a past admixture of individuals from discrete populations of the same species and interspecific hybridisation not only between V. myrtillus and V. vitis-idaea but also between V. uliginosum and V. vitis-idaea, the latter not being reported to date. Conservation measures suitable for preservation of presumably genetically distinct portions of the Balkans' gene pools of studied species have been suggested, while the utility of interspecific hybrids in breeding programs and/or in food/pharmaceutical industry is yet to be assessed

Comparative studies on osmosis based encapsulation of sodium diclofenac in porcine and outdated human erythrocyte ghosts

The objective of our study was to develop controlled drug delivery system based on erythrocyte ghosts for amphiphilic compound sodium diclofenac considering the differences between erythrocytes derived from two readily available materials - porcine slaughterhouse and outdated transfusion human blood. Starting erythrocytes, empty erythrocyte ghosts and diclofenac loaded ghosts were compared in terms of the encapsulation efficiency, drug releasing profiles, size distribution, surface charge, conductivity, surface roughness and morphology. The encapsulation of sodium diclofenac was performed by an osmosis based process - gradual hemolysis. During this process sodium diclofenac exerted mild and delayed antihemolytic effect and increased potassium efflux in porcine but not in outdated human erythrocytes. FTIR spectra revealed lack of any membrane lipid disorder and chemical reaction with sodium diclofenac in encapsulated ghosts. Outdated human erythrocyte ghosts with detected nanoscale damages and reduced ability to shrink had encapsulation efficiency of only 8%. On the other hand, porcine erythrocyte ghosts had encapsulation efficiency of 37% and relatively slow drug release rate. More preserved structure and functional properties of porcine erythrocytes related to their superior encapsulation and release performances, define them as more appropriate for the usage in sodium diclofenac encapsulation process

Anti-encephalitogenic effects of cucumber leaf extract

Cucumber (Cucumis sativus) fruit has been used in cuisine worldwide, while its leaves are rich in immunomodulatory compounds. Cucumber leaf extract (CLE) was characterized by the predominance of triterpenoids cucurbitacins and significant levels of phenolics. Effects of CLE on CD4(+) T helper (Th) cells and macrophages, as the major encephalitogenic cells in the autoimmunity of the central nervous system were investigated in our study. CLE potently inhibited production of major pathogenic Th cytokines: interferon-gamma and interleukin-17, as well as of nitric oxide and reactive oxygen species in macrophages. Antigen-presenting activity of macrophages and dendritic cells was also affected by CLE. The effects of CLE were co-incident with modulation of NFKB and p38 MAPK signaling. Concentrations of CLE used in vitro did not show toxic effects on zebrafish embryos. Moreover, CLE inhibited generation of encephalitogenic cells in vivo. These results demonstrate that CLE deserve further investigation on its anti-encephalitogenic therapeutic properties

Trichome-specific and developmentally regulated biosynthesis of nepetalactones in leaves of cultivated Nepeta rtanjensis plants

Nepetalactones are accumulated as the main secondary metabolites in the majority of Nepeta species; however, their biosynthetic pathway remains to a great extent unknown, as well as their relationship to the overall physiology of these plants. Nepetalactones most certainly arise through general iridoid biosynthetic pathway leading to nepetalactol, a common precursor of all plant-derived iridoids that is formed in a reaction catalysed by iridoid synthase (IS, EC 1.3.1.99). In this paper, we aimed to localize the sites of nepetalactone biosynthesis and accumulation by comparatively analysing the nepetalactones content and the expression of putative iridoid synthase (NrIS) in leaves of Nepeta rtanjensis Diklic & Milojevic, a trans,cis-nepetalactone and dehydronepetalactone rich species. Another aim was to give an insight into the developmental regulation of nepetalactone biosynthesis in leaves of cultivated N. rtanjensis plants. Knowledge on the spatial-, and developmental regulation of nepetalactone metabolism could greatly benefit potential growers by enabling them to adequately plan and manage plant cultivation, harvesting, and handling and thus increase yield of bioactive compounds. The expression profile of putative NrIS1 is highly correlated with trans,cis-nepetalactone and dehydronepetalactone content/ratio in leaves of different developmental stages. The results strongly indicate that developmental regulation of nepetalactone biosynthesis in leaves resides largely at the level of gene expression, and suggest an important role of NrISI. Young leaves exhibited the most intensive nepetalactone biosynthesis and accumulation, but also its conversion to dehydronepetalactone. Thus, the pattern of NL accumulation in leaves can be explained by both the rate of its biosynthesis and metabolic degradation. The gene expression patterns of NrIS1 and NrIS2 transcripts in trichomes and abraded leaves revealed trichomes as the main site of their activity. UHPLC/qqqMS profiling of nepetalactones in trichomes and abraded leaves, complemented with Raman spectroscopy and dichloromethane dipping experiment, supported glandular trichome-specific localization of nepetalactone biosynthesis and accumulation

Nepeta rtanjensis (Lamiaceae), a plant endemic to the Balkans: Phenolic composition antioxidant activity, and in vitro antigenotoxic effects in triiodothyronine-induced DNA damage in human lymphocytes

The success of antioxidant therapy in hyperthyroidism implies that disease is mediated by oxidative stress, which is known as one of the causing agents of agemg, degenerative diseases, and cancer. The main objective of our study was to determine possible protective effects of methanolic extract of N. rtanjensis in triiodothyronine (T-3)-induced DNA breaks of human lymphocytes under in vitro conditions, based upon plant antioxidant capacity related to its phytochemical profile, mainly its polyphenolic complex. The total phenolic and flavonoid content and the antioxidant activity using in vitro 1,1-dyphenyl-2-picrylhydrazyl reagent (DPPH) was determined in methanolic extracts of plant leaves and flowers. The phenolic compound content of 62.73 +/- 1.80 mg of GaA/g, exhibited solid antioxidant activity (IC50= 112.59 +/- 0.95 g/ml). The antigenotoxic activity of 0.2, 0.5 and 1.0 mg/ml N. rtanjensis methanol extracts mixture with 100 mu M of T-3 was studied in human lymphocytes in vitro using the Comet assay. It is supposed that the antigenotoxicity of N. rtanjensis methanol extracts was caused by high presence of chlorogenic acid, rosmarinic acid and rutin, all known as efficient antioxidant bioactive compounds, which were determined by ultrahigh-pressure liquid chromatograph with MS/MS Mass Spectroscopy (UHPLC/-HESI-MS/MS)

Nepeta rtanjensis (Lamiaceae), a plant endemic to the Balkans: Phenolic composition antioxidant activity, and in vitro antigenotoxic effects in triiodothyronine-induced DNA damage in human lymphocytes

The success of antioxidant therapy in hyperthyroidism implies that disease is mediated by oxidative stress, which is known as one of the causing agents of agemg, degenerative diseases, and cancer. The main objective of our study was to determine possible protective effects of methanolic extract of N. rtanjensis in triiodothyronine (T-3)-induced DNA breaks of human lymphocytes under in vitro conditions, based upon plant antioxidant capacity related to its phytochemical profile, mainly its polyphenolic complex. The total phenolic and flavonoid content and the antioxidant activity using in vitro 1,1-dyphenyl-2-picrylhydrazyl reagent (DPPH) was determined in methanolic extracts of plant leaves and flowers. The phenolic compound content of 62.73 +/- 1.80 mg of GaA/g, exhibited solid antioxidant activity (IC50= 112.59 +/- 0.95 g/ml). The antigenotoxic activity of 0.2, 0.5 and 1.0 mg/ml N. rtanjensis methanol extracts mixture with 100 mu M of T-3 was studied in human lymphocytes in vitro using the Comet assay. It is supposed that the antigenotoxicity of N. rtanjensis methanol extracts was caused by high presence of chlorogenic acid, rosmarinic acid and rutin, all known as efficient antioxidant bioactive compounds, which were determined by ultrahigh-pressure liquid chromatograph with MS/MS Mass Spectroscopy (UHPLC/-HESI-MS/MS)