Electroporation Enhances the Anticancer Effects of Novel Cu(II) and Fe(II) Complexes in Chemotherapy-Resistant Glioblastoma Cancer Cells.

Schiff base ligand (L) was obtained by condensation reaction between 4-aminopyrimidin-2(1H)-one (cytosine) with 2-hydroxybenzaldehyde. The synthesized Schiff base was used for complexation with Cu(II) and Fe(II) ions used by a molar (2 : 1 mmol ration) in methanol solvent. The structural features of ligand, Cu(II), and Fe(II) metal complexes were determined by standard spectroscopic methods (FT-IR, elemental analysis, proton and carbon NMR spectra, UV/VIS, and mass spectroscopy, magnetic susceptibility, thermal analysis, and powder X-ray diffraction). The synthesized compounds (Schiff base and its metal complexes) were screened in terms of their anti-proliferative activities in U118 and T98G human glioblastoma cell lines alone or in combination with electroporation (EP). Moreover, the human HDF (human dermal fibroblast) cell lines was used to check the bio-compatibility of the compounds. Anti-proliferative activities of all compounds were ascertained using an MTT assay. The complexes exhibited a good anti-proliferative effect on U118 and T98G glioblastoma cell lines. In addition, these compounds had a negligible cytotoxic effect on the fibroblast HDF cell lines. The use of compounds in combination with EP significantly decreased the IC50 values compared to the use of compounds alone (p<0.05). These results show that newly synthesized Cu(II) and Fe(II) complexes can be developed for use in the treatment of chemotherapy-resistant U118 and T98G glioblastoma cells and that treatment with lower doses can be provided when used in combination with EP.

Synthesis, characterization, antioxidant and anticancer activities of a new Schiff base and its M(II) complexes derived from 5-fluorouracil.

In this study, Schiff base ligand was obtained from the condensation reaction of benzene-1,2-diamine and 5-fluoropyrimidine-2,4(1H,3H)-dione (5-FU). Metal(II) complexes were synthesized with Fe(II), Co(II) and Ni(II) chloride salts. The synthesized ligand and metal complexes were characterized by FT-IR, UV-vis, 1H-13C NMR, elemental analyses, mass spectroscopy, magnetic moments, molar conductivity and thermogravimetric analysis studies. With the help of different techniques reveal Fe(II), Co(II) and Ni(II) complexes have exhibited tetrahedral and octahedral geometry. Ligand acted as bidentate and it binds metal(II) ions through deprotonated-NH, imine-N atom and carbonyl-O atom, respectively. DPPH, ABTS, FRAP, CUPRAC and total antioxidant activity methods were used to determine the antioxidant properties of ligand and metal complexes. According to the results, the synthesized compounds showed very high antioxidant activity compared to 5-FU. The cytotoxicities of the synthesized compounds were performed on MCF-7 (human breast cancer) and L-929 (fibroblast) cell lines using the MTT assay. In addition, the effect of electroporation (EP) on the cytotoxicity of the compounds was investigated. Our results demonstrated that novel Co(II) and Ni(II) complexes show potential as new anticancer agents and ECT may be a viable treatment option for breast cancer.

Postbiotic metabolites, antioxidant and anticancer activities of probiotic Leuconostoc pseudomesenteroides strains in natural pickles.

In this study, five strains of Leuconostoc pseudomesenteroides were thought to have probiotic properties and anticancer activity isolated from natural pickles and identified by performing the 16S rRNA sequence analysis. The probiotic properties, postbiotic amounts, the capacity to adhere to the L-929, HT-29 and Caco-2 cell lines, the effects of postbiotic and bacterial extracts on cell viability and biochemical activities were investigated in the strains. In the results, Leu. pseudomesenteroides Y6 strain was detected to have the best resistance to the stomach and intestinal environments, and the quantities of postbiotic metabolites are similar to each other. The bacterial adhesion capacities were found to be in the range of 1.66-8.5%. Furthermore, postbiotic metabolites of all isolates had good anticancer activity (27.67-86.05%) and the activity of bacterial extractions increased depending on concentration. Leu. pseudomesenteroides Y4 and Y6 strains generally showed better activity than controls and all strains were strong 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavengers in the antioxidant studies. In conclusion, the Y6 strain, which had the best probiotic feature, was found to show significantly good biological activity. It is thought that this isolate will be supported by new in vivo studies and eventually be brought to the food and health industry.

Preparation of Cu(II), Ni(II), Ti(IV), VO(IV), and Zn(II) Metal Complexes Derived from Novel vic-Dioxime and Investigation of Their Antioxidant and Antibacterial Activities.

In this work, novel vic-dioxime ligand (LH2 ) containing bound to the N4 -oxime core moiety and its complexes with Cu(II), Ni(II), Ti(IV), VO(IV), and Zn(II) salts have been studied. The structure of the ligand and its complexes were successfully synthesized and characterized using NMR (1 H and 13 C), LC/MS/MS spectrometer, FT-IR and UV/VIS spectroscopy, melting point, and magnetic susceptibility measurements. Vic-dioxime ligand (LH2 ) (1) and its metal complexes ([Cu(LH)2 ] (2), [Ni(LH)2 ] (3), [Ti(LH)2 ]Cl2 (4), [VO(LH)2 ] (5), and [Zn(LH)2 ] (6), respectively) were tested for them in-vitro antibacterial and antioxidant activities. According to the metal chelating results of the study, it was determined that compounds (1), (2), (3), and (6) showed very good activity, and especially compound (2), had a stronger metal chelating capacity due to ligand dissociation from the synthesized metal complexes, which then would chelate Fe(II) in the experimental setting. When microorganisms were evaluated in terms of the % viability effect, it was observed that all compounds had activity against C. Albicans and S. Cerevisiae at rates similar to antibiotics.

Synthesis, characterization, antiproliferative of pyrimidine based ligand and its Ni(II) and Pd(II) complexes and effectiveness of electroporation.

In the study, a new Schiff base (ligand) was obtained using 4-aminopyrimidine-2(1H)-one, the starting material, and 2,3,4-trimethoxy benzaldehyde. Ni(II) and Pd(II) complexes were obtained from the reaction of the ligand and NiCl2·6H2O, PdCl2(CH3CN)2 (1:1 ratio). These compounds were characterized using the elemental and mass analysis, 1H, 13C-NMR, FT-IR, UV-Vis, magnetic susceptibility, thermal analysis, and the X-ray diffraction analyses. The antiproliferative activities of the synthesized ligand, Ni(II) and Pd(II) complexes were identified on the HepG2 (human liver cancer cells) cell line and their biocompatibility was tested on the L-929 (fibroblast cells) cell line by the MTT analysis method. Furthermore, the effects of electroporation (EP) on the cytotoxic activities of synthesized compounds were investigated in HepG2 cancer cells. According to the MTT findings of the study, the ligand did not exhibit an antiproliferative activity while its Ni(II) and Pd(II) complexes exhibited an antiproliferative activity. Moreover, it was observed that the antiproliferative activity of the Pd(II) complex was stronger than that of the Ni(II) complex. The combined application of EP + compounds is much more effective than the usage of the compounds alone in the treatment of HepG2 cancer cells. The EP increased the cytotoxicity of the Ni(II) and Pd(II) complexes by 1.66, and 2.54 times, respectively. It was concluded that Ni(II) and Pd(II) complexes may contribute as potential anti-cancer agents for the treatment of hepatocellular carcinoma and yield promising results in the case of being used in ECT.Communicated by Ramaswamy H. Sarma.

Effects of electroporation on anticancer activity of 5-FU and newly synthesized zinc(II) complex in chemotherapy-resistance human brain tumor cells.

Zn(II) complex of Schiff base derived from the condensation of 4-aminopyrimidine-2(1H)-one with salicylaldehyde was prepared and characterized by various physico-chemical and spectral methods for structure determination. The cytotoxic activity of the Zn(II) complex was investigated in comparison with 5-fluorouracil (5-FU) against two different human brain tumor cell lines (T98G and U118), while primer human dermal fibroblast cells (HDF) was used as control for biocompatibility. Then, the effectiveness of electroporation (EP) on cytotoxic activities of these compounds has been examined. The cytotoxicities of the 5-FU and new Zn(II) complex, alone or in combination with electroporation, were determined by MTT assay. The Zn(II) complex showed good cytotoxicity against T98G and U118 brain tumor cell lines with IC50 = 282.47 and 297.91 µM respectively, while it was safe on HDF healthy cells with IC50 = 826.72 µM. The 5-FU exhibited less cytotoxicity compared to the Zn(II) complex against T98G (IC50 = 382.35 µM) and U118 (IC50 = 396.56 µM) tumor cell lines. The combined application of Zn (II) + EP decreased the IC50 value by 5.96-fold in T98G cells and 4.76-fold in U118 cells. EP showed a similar effect in its combined application with 5-FU, resulting in a decrease of the IC50 value of 4.22-fold in the T98G cells and 3.84-fold in the U118 cells. In a conclusion, the Zn(II) complex exhibited an anticancer potential against both brain tumor cell lines (T98G and U118) and EP greatly increased the cytotoxicity of Zn(II) complex and 5-FU on these chemotherapy-resistant cells.

Culture fermentation of Lactobacillus in traditional pickled gherkins: Microbial development, chemical, biogenic amine and metabolite analysis.

Fermented cucumber pickles are the lactic acid fermentation products formed through the influence of microorganisms present in the environment. This study investigated the impacts of starter cultures, namely, Lactobacillus plantarum, Lactobacillus pentosus and Lactobacillus paraplantarum, typically utilized for the fermentation of traditional pickled gherkins, on fermentation process. The chemical (pH, total acidity and salt) and microbiological (total mesophilic aerobic bacteria, lactic acid bacteria and yeast-mould) changes were observed against the control sample during fermentation process. Moreover, the amounts of biogenic amines (BAs) and metabolites formed as a consequence of fermentation were determined using HPLC. It was found that the chemical analyses provided similar results for all the samples. The amount of total mesophilic aerobic bacteria and yeast-mould colonies in pickle sample containing L. plantarum 49 strain appeared to reduce significantly. The amount of BAs was the lowest for the pickle samples where L. plantarum strains were added. The amount of BAs was below the toxic value that could affect human health. More BAs were synthesized as the fermentation period increased. Lactate was seen to exist in the samples when pyruvate was present, and acetoin was converted into 2.3-butanediol during the fermentation period. It was concluded that the pickle sample for which L. plantarum 49 strain was used displayed a better fermentation profile (i.e., metabolite and biogenic amines) than the remaining samples. Producing a more delicious and reliable product using such characteristics of L. plantarum strains in pickled gherkins is believed to significantly contribute to the food industry.

Assessment of Antimicrobial and Antioxidant Activities of Nepeta trachonitica: Analysis of Its Phenolic Compounds Using HPLC-MS/MS.

Continuing our work on the sources of natural bioactive compounds, we evaluated the antimicrobial and antioxidant activities of Nepeta trachonitica as well as its major phenolic content using the high-performance liquid chromatography-mass spectrometry/mass spectrometry (HPLC-MS/MS) technique. For antioxidant activity, ferric reducing antioxidant power (FRAP) and cupric ion reducing antioxidant capacity (CUPRAC) methods were performed to measure the reducing power and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay was employed to evaluate the radical scavenging activity of the sample. For antimicrobial activity, three Gram-positive and four Gram-negative microbial species as well as three fungi species were tested. N. trachonitica appeared to have reasonable antioxidant activity and decent antimicrobial activity as indicated by the inhibition of the organisms' growth. The most susceptible species were Bacillus subtilis ATCC 6633 and Escherichia coli ATCC 11229 among the organisms tested. Ethanol extract of the plant has the highest effect on Saccharomyces cerevisiae but no effect on Yarrowia lipolytica. The HPLC-MS/MS analysis showed that at least 11 major phenolic compounds of N. trachonitica exist, the major ones being rosmarinic acid, chlorogenic acid and quinic acid. The obtained results suggest that N. trachonitica could be a promising source for food and nutraceutical industries because of its antimicrobial and antioxidant properties and phenolic compounds.

Enhanced electrokinetic properties and antimicrobial activities of biodegradable chitosan/organo-bentonite composites.

In this study, chitosan (CS), Na+-bentonite (Na+-BNT) and chitosan/organo-bentonite (CS/O-BNT) biodegradable composites having three different compositions were investigated. Electrokinetic measurements were examined in aqueous medium by taking the effects pH, electrolytes (NaCl and BaCl2), surfactants (CTAB and SDS), and temperature into account. It was noticed that the initial ζ-potential of Na+-BNT shifted from negative (ζ=-35mV) to positive region (ζ=+13mV) with increasing polycationic CS content in the composite structure as aimed. Divalent 2:1 electrolyte (BaCl2) caused to shift the ζ-potentials of all the dispersions to more positive regions. While the most negative effect on ζ-potential of the composites was reached with SDS, which reduced the value of ζ-potential to -39mV for CS(1)/O-BNT composite, the most positive effect was monitored with CTAB (ζ=+40mV) for CS(3)/O-BNT composite. Further, the composites were tested against various bacterial (Gram-positive and Gram-negative) and fungal microorganisms at various concentrations and results obtained were compared with the reference antibiotics and fungicide. According to inhibition zone values accomplished, antibacterial and antifungal activities of the CS/O-BNT composites are increased with increasing CS content as proportional with their positive ζ-potential values.

RP-HPLC/MS/MS Analysis of the Phenolic Compounds, Antioxidant and Antimicrobial Activities of Salvia L. Species.

The identification and quantification of the phenolic contents of methanolic extracts of three Salvia L. species namely S. brachyantha (Bordz.) Pobed, S. aethiopis L., and S. microstegia Boiss. and Bal. were evaluated using reverse phase high performance liquid chromatography, UV adsorption, and mass spectrometry (RP-HPLC/MS). In order to determine the antioxidant capacity of these species, cupric ions (Cu2+) reducing assay (CUPRAC) and ferric ions (Fe3+) reducing assay (FRAP) were performed to screen the reducing capacity and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay was employed for evaluation of the radical scavenging activity for both solvents. In further investigation, the antimicrobial activities of Salvia species were tested using the disc diffusion method against three Gram-positive and four Gram-negative microbial species, as well as three fungi species. The results showed that there is a total of 18 detectable phenols, the most abundant of which was kaempferol in S. microstegia and rosmarinic acids in S. brachyantha and S aethiopis. The other major phenols were found to be apigenin, luteolin, p-coumaric acid, and chlorogenic acid. All species tested showed moderate and lower antioxidant activity than standard antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and ascorbic acid. The ethanolic extracts of Salvia species revealed a wide range of antimicrobial activity. S. brachyantha and S. microstegia showed the highest antimicrobial activities against B. subtilis, whereas S. aethiopis was more effective on Y. lipolytica. None of the extracts showed anti-fungal activity against S. cerevisiae. Thus these species could be valuable due to their bioactive compounds.

Detection of putative virulence genes in Aeromonas isolates from humans and animals.

INTRODUCTION: Aeromonas are food- and water-borne bacteria that are considered to be zoonotic human pathogens. This study aimed to investigate the presence of genes associated with virulence in human and animal Aeromonas isolates and the potential role of animal isolates with regards to human Aeromonas infections. METHODOLOGY: The presence of aerA, hlyA, alt, ast, laf, ascF-G, stx1 and stx2 putative virulence genes in 40 human and animal Aeromonas isolates (16 human and 24 animal isolates) were examined by polymerase chain reaction (PCR). DNA fragments of expected sizes were purified and sequenced. BLAST in the NCBI was used to verify any amplified products. RESULTS: PCR screening showed that hlyA, alt, and laf genes were determined at ratios of 6.25%, 50%, and 6.25%, respectively, in human isolates. The ratios of hlyA, alt, ascF-G, laf, stx2, and stx1 genes in animal isolates were 58.3%, 20.83%, 33.3%, 20.83%, 8.33%, and 4.17%, respectively. Neither aerA nor ast genes were detected in any isolates. Any one of eight putative virulence genes was not detected in seven human and eight animal isolates in the study. CONCLUSIONS: The current study is the first to investigate the presence of the virulence gene in gull Aeromonas isolates. The manifestation of the presence of the virulence gene and gene combinations was considerable, especially in fish and gull isolates when compared with clinical human isolates. The current study demonstrates the potential importance of fish and gulls in terms of human Aeromonas infections.

The synthesis, characterization, antimicrobial and antimutagenic activities of hydroxyphenylimino ligands and their metal complexes of usnic acid isolated from Usnea longissima.

Novel multifunctional hydroxyphenylimino ligands (L1, L2 and L3) were synthesized by the condensation of 2-aminophenol, 3-aminophenol and 4-aminophenol with usnic acid, a lichen metabolite. The synthesized ligands and their Cu(II), Co(II), Ni(II) and Mn(II) complexes were characterized using FT-IR, UV-Vis, (1)H-NMR, (13)C-NMR, 1D- and 2D NMR (DEPT, COSY, HMQC and HMBC), LC-MS and TGA. In addition, the metal complexes of the novel ligands were prepared with high yields using Cu(II), Co(II), Ni(II) and Mn(II) salts and were characterized using the FT-MIR/FAR, UV-Vis, elemental analysis, ICP-OES and TG/DTA techniques. The ligands and their complexes were tested against ten important pathogen microorganisms using the disc diffusion method and the metal complexes of the ligands were more active against all of the microorganisms tested with a broad spectrum than the ligands exhibiting 11­32 mm inhibition zones. On the other hand, a broad spectrum of the strongest antimicrobial activity was determined for the Mn(II) and Cu(II) complexes of the hydroxyphenylimino ligand with usnic acid (L3). In addition, the antimutagenic activities of all of the ligands and their metal complexes were determined using the Ames-Salmonella and E. coli WP2 microbial assay systems and they showed varied and strong antimutagenic effects. In general, it has been found that the Co and Mn complexes of the ligands possess potent antimutagenic activity. In view of these results, it can be concluded that some metal complexes can be used as antimicrobial and anticancer agents.