Antioxidant, antigenotoxic and cytotoxic activity of essential oils and methanol extracts of hyssopus officinalis l. Subsp. aristatus (godr.) nyman (lamiaceae)

Abstract:Hyssopus officinalisL. is a well-known aromatic plant used in traditional medicine and the foodand cosmetics industry. The aim of this study is to assess the antioxidant, genotoxic, antigenotoxic andcytotoxic properties of characterized hyssop essential oils and methanol extracts. Chemical compositionwas analyzed by gas chromatography - mass spectrometry (GC-MS) and liquid chromatography with diodearray detection and mass spectrometry (LC-DAD-MS), respectively. Antioxidant activity was examinedby 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing/antioxidant power (FRAP) tests; genotoxicand antigenotoxic activity were examined by the comet assay, while cytotoxicity was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide dye (MTT) test against tumor cell lines (SW480,MDA-MB 231, HeLa) and non-transformed human lung fibroblast cell lines (MRC-5). The essential oilswere rich in monoterpene hydrocarbons (e.g., limonene; 7.99–23.81%), oxygenated monoterpenes (1,8-cineole; 38.19–67.1%) and phenylpropanoids (methyl eugenol; 0.00–28.33%). In methanol extracts, the mostabundant phenolics were chlorogenic and rosmarinic acid (23.35–33.46 and 3.53–17.98 mg/g, respectively).Methanol extracts expressed moderate to weak antioxidant activity (DPPH IC50= 56.04–199.89μg/mL,FRAP = 0.667–0.959 mmol Fe2+/g). Hyssop preparations significantly reduced DNA damage in humanwhole blood cells, induced by pretreatment with hydrogen peroxide. Methanol extracts exhibited selectiveand potent dose- and time-dependent activity against the HeLa cell line. Results of the current studydemonstrated notableH.officinalismedicinal potential, which calls for further investigation

Antiproliferative and Proapoptotic Activities of Methanolic Extracts from Ligustrum vulgare L. as an Individual Treatment and in Combination with Palladium Complex

The aim of this study is to examine the growth inhibitory effects of methanolic leaf and fruit extracts of L. vulgare on HCT-116 cells over different time periods and their synergistic effect with a Pd(apox) complex. The antiproliferative activity of plant extracts alone or in combination with the Pd(apox) complex was determined using MTT cell viability assay, where the IC50 value was used as a parameter of cytotoxicity. Results show that antiproliferative effects of L. vulgare extracts increase with extension of exposure time, with decreasing IC50 values, except for 72 h where the IC50 values for methanolic leaf extract were lower than for the fruit extract. The Pd(apox) complex alone had a weak antiproliferative effect, but combination with L. vulgare extracts caused stronger effects with lower IC50 values than with L. vulgare extracts alone. The type of cell death was explored by fluorescence microscopy using the acridin orange/ethidium bromide method. Treatments with plant extracts caused typical apoptotic morphological changes in HCT-116 cells and co-treatments with Pd(apox) complex caused higher levels of apoptotic cells than treatment with plant extracts alone. The results indicate that L. vulgare is a considerable source of natural bioactive substances with antiproliferative activity on HCT-116 cells and which have a substantial synergistic effect with the Pd(apox) complex

Antioxidant, Antimicrobial and Antiproliferative Activities of Five Lichen Species

The antioxidative, antimicrobial and antiproliferative potentials of the methanol extracts of the lichen species Parmelia sulcata, Flavoparmelia caperata, Evernia prunastri, Hypogymnia physodes and Cladonia foliacea were evaluated. The total phenolic content of the tested extracts varied from 78.12 to 141.59 mg of gallic acid equivalent (GA)/g of extract and the total flavonoid content from 20.14 to 44.43 mg of rutin equivalent (Ru)/g of extract. The antioxidant capacities of the lichen extracts were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging. Hypogymnia physodes with the highest phenolic content showed the strongest DPPH radical scavenging effect. Further, the antimicrobial potential of the lichen extracts was determined by a microdilution method on 29 microorganisms, including 15 strains of bacteria, 10 species of filamentous fungi and 4 yeast species. A high antimicrobial activity of all the tested extracts was observed with more potent inhibitory effects on the growth of Gram (+) bacteria. The highest antimicrobial activity among lichens was demonstrated by Hypogymnia physodes and Cladonia foliacea. Finally, the antiproliferative activity of the lichen extracts was explored on the colon cancer adenocarcinoma cell line HCT-116 by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) viability assay and acridine orange/ethidium bromide staining. The methanol extracts of Hypogymnia physodes and Cladonia foliacea showed a better cytotoxic activity than the other extracts. All lichen species showed the ability to induce apoptosis of HCT-116 cells

Cadmium-Induced Oxidative Stress and Apoptotic Changes in the Testis of Freshwater Crab, Sinopotamon henanense

Cadmium (Cd), one of the most toxic environmental and industrial pollutants, is known to exert gonadotoxic and spermiotoxic effects. In the present study, we examined the toxic effect of Cd on the testis of freshwater crab, Sinopotamon henanense. Crabs were exposed to different Cd concentrations (from 0 to 116.00 mg·L−1) for 7 d. Oxidative stress and apoptotic changes in the testes were detected. The activities of SOD, GPx and CAT initially increased and subsequently decreased with increasing Cd concentrations, which was accompanied with the increase in malondialdehyde (MDA) and H2O2 content in a concentration-dependent manner. Typical morphological characteristic and physiological changes of apoptosis were observed using a variety of methods (HE staining, AO/EB double fluorescent staining, Transmission Electron Microscope observation and DNA fragmentation analysis), and the activities of caspase-3 and caspase-9 were increased in a concentration-dependent manner after Cd exposure. These results led to the conclusion that Cd could induced oxidative damage as well as apoptosis in the testis, and the apoptotic processes may be mediated via mitochondria-dependent apoptosis pathway by regulating the activities of caspase-3 and caspase-9

Macromolecular design of folic acid functionalized amylopectin- albumin core-shell nanogels for improved physiological stability and colon cancer cell targeted delivery of curcumin

Nanogels have potential for encapsulating cancer therapeutics, yet their susceptibility to physiological degradation and lack of cellular specificity hinder their use as effective oral delivery vehicles. Herein, we engineered novel albumin-core with folic acid functionalized hyperbranched amylopectin shell-type nanogels, prepared through a two-step reaction and loaded with curcumin while the proteinaceous core was undergoing thermal gelation. The nanogels had a mean hydrodynamic diameter of ca. 90 nm and ζ-potential of ca. -24 mV. Encapsulation of curcumin within the nanogels was restored, up to ca. 0.05 mg mL-1, beyond which, a gradual increase in size and a decrease in ζ-potential was observed. The core-shell structures were resilient to in vitro physiological oral-gastrointestinal digestion owing to a liquid crystalline B- and V-type polymorphism in the polysaccharide shell, the latter being driven by the shell functionalization with folic acid. Additionally, these biocompatible nanogels restored stability of the encapsulated curcumin and exhibited augmented cellular uptake and retention specifically in folate receptor-positive HT29 human colon adenocarcinoma cells, inducing early-stage apoptosis. Novel insights from this study represent a promising platform for rational designing of future oral delivery systems that can surmount physiological barriers for delivering cancer therapeutics to colon cancer cells with improved stability and specificity

Prevalence of Genotypes That Determine Resistance of Staphylococci to Macrolides and Lincosamides in Serbia

Macrolides, lincosamides, and streptogramins (MLS) resistance genes are responsible for resistance to these antibiotics in Staphylococcus infections. The purpose of the study was to analyze the distribution of the MLS resistance genes in community- and hospital-acquired Staphylococcus isolates. The MLS resistance phenotypes [constitutive resistance to macrolide–lincosamide–streptogramin B (cMLSb), inducible resistance to macrolide–lincosamide–streptogramin B (iMLSb), resistance to macrolide/macrolide–streptogramin B (M/MSb), and resistance to lincosamide–streptogramin A/streptogramin B (LSa/b)] were determined by double-disc diffusion method. The presence of the MLS resistance genes (ermA, ermB, ermC, msrA/B, lnuA, lnuB, and lsaA) were determined by end-point polymerase chain reaction in 179 isolates of staphylococci collected during 1-year period at the Center for Microbiology of Public Health Institute in Vranje. The most frequent MLS phenotype among staphylococcal isolates, both community-acquired and hospital-acquired, was iMLSb (33.4%). The second most frequent was M/MSb (17.6%) with statistically significantly higher number of hospital-acquired staphylococcal isolates (p < 0.05). MLS resistance was mostly determined by the presence of msrA/B (35.0%) and ermC (20.8%) genes. Examined phenotypes were mostly determined by the presence of one gene, especially by msrA/B (26.3%) and ermC (14.5%), but 15.6% was determined by a combination of two or more genes. M/MSb phenotype was the most frequently encoded by msrA/B (95.6%) gene, LSa/b phenotype by lnuA (56.3%) gene, and iMLSb phenotype by ermC (29.4%) and ermA (25.5%) genes. Although cMLSb phenotype was mostly determined by the presence of ermC (28.9%), combinations of two or more genes have been present too. This pattern was particularly recorded in methicillin-resistant Staphylococcus aureus (MRSA) (58.3%) and methicillin-resistant coagulase-negative staphylococci (MRCNS) (90.9%) isolates with cMLSB phenotype. The msrA/B gene and M/MSb phenotype were statistically significantly higher in hospital-acquired than community-acquired staphylococci strains (p < 0.05). There are no statistically significant differences between staphylococci harboring the rest of MLS resistance genes acquired in community and hospital settings (p > 0.05). The prevalence of iMLSb phenotypes may change over time, so it is necessary to perform periodic survey of MLS resistance phenotypes, particularly where the D-test is not performed routinely

The cytotoxicity of korbazol against murine cancer cell lines

Background/aim: Korbazol is a natural product that has been shown to exert cytotoxic eff ects on leukemic cells in vitro. The cytotoxicity and biochemical eff ects induced by Korbazol were investigated in the murine cell lines 4T1, B16 and BCL1. Methods: The cytotoxic activity of the tested compound was assessed using a colorimetric MTT assay. The concentration of the superoxide anion radical (O 2.-) and the activity of superoxide dismutase (SOD) in the samples were determined using spectrophotometric methods. The MDA content was determined using a TBA assay. Results: We found that Korbazol induced cell toxicity, an increased the concentration of the lipid peroxidation end product MDA, decreased superoxide dismutase activity and increased superoxide anion formation. Conclusions: Altogether, these results suggest that oxidative stress is involved in Korbazol-induced cytotoxicity in the investigated cell lines

Effects of host defense peptides B2RP, Brevinin-2GU, D-Lys-Temporin, Lys-XT-7 and D-Lys-Ascaphin-8 on peripheral blood mononuclear cells: Preliminary study

© 2017 Croatian Society of Natural Sciences. All rights reserved. Background and purpose: Host defense peptides have considerable therapeutic potential. One of the limitations for their therapeutic use is insufficient selectivity of some peptides, i.e. toxicity for eukaryotic cells. In this study, we have investigated effect of two naturally occurring and three analogs of frog skin-derived peptides on viability/proliferation of resting peripheral blood mononuclear cells and activated lymphocytes. Materials and Methods: Effect of tested peptides was assessed using MTT colorimetric assay. Concanavalin A was used as lymphocyte mitogen. Results: Brevinin-2GU induced cell death only in the highest tested concentration, whereas other peptides were not cytotoxic to resting peripheral blood mononuclear cells. Moreover, high concentrations of B2RP, DLys-Ascaphin-8 and Lys-XT-7 induced cell proliferation and this effect was more prominent in lymphocytes (p<0.05). Tested peptides had opposite effect on activated lymphocytes inhibiting proliferative response to Concanavalin A (Brevinin-2GU, B2RP and D-Lys-Temporin p<0.05). Conclusions: Tested peptides (with exception of Brevinin-2GU) didn’t show cytotoxicity toward peripheral blood mononuclear cells. Moreover, they have potential to modulate immune response by inducing proliferation of resting peripheral blood mononuclear cells and limiting proliferative response to the activation stimulus. Regarding their potent antimicrobial and low hemolytic activity this makes them good candidates for therapeutic use

Suppression of natural killer-cell and dendritic-cell apoptotic tumoricidal activity in patients with head and neck cancer

Background Natural killer (NK) cells and dendritic cells (DCs) mediate tumor cell apoptosis using tumor necrosis factor superfamily ligands (TNFSFLs). This cytotoxicity is an important anticancer immune defense mechanism. Methods We examined TNFSFL expression and apoptotic tumoricidal activity (ATA) of purified NK cells and DCs, and peripheral blood mononuclear leukocytes (PBMLs) of healthy individuals and patients with head and neck cancer (HNC) before and after cancer ablation. Results PBMLs, NK cells and DCs, but not NK-cell/DC-depleted PBMLs, expressed multiple TNFSFLs and mediated ATA. Both TNFSFL expression and ATA were suppressed in tumor-bearing, and restored in tumor-ablated patients with (HNC) Soluble TNF superfamily receptors (solTNFSFRs) were increasingly bound by PBNLs of tumor-bearing HNC patients. Dissociation of solTNFSFR led to more pronounced increases in TNFSFL expression and ATA of PBMLs of patients with HNC than healthy individuals. Conclusion NK-cell and DC TNFSFL expression and ATA are suppressed in patients with HNC. This suppression is tumor-dependent and possibly mediated by solTNFSFRs. © 2012 Wiley Periodicals, Inc