Newly synthesized palladium(II) complexes with dialkyl esters of (S,S)-propylenediamine-N,N'-di-(2,2'-di-(4-hydroxy-benzil))acetic acid: in vitro investigation of biological activities and HSA/DNA binding.

The four new ligands, dialkyl esters of (S,S)-propylenediamine-N,N'-di-(2,2'-di-(4-hydroxy-benzil))acetic acid (R2-S,S-pddtyr·2HCl) (R = ethyl (L1), propyl (L2), butyl (L3), and pentyl (L4)) and corresponding palladium(II) complexes have been synthesized and characterized by microanalysis, infrared, 1H NMR and 13C NMR spectroscopy. In vitro cytotoxicity was evaluated using the MTT assay on four tumor cell lines, including mouse mammary (4T1) and colon (CT26), and human mammary (MDA-MD-468) and colon (HCT116), as well as non-tumor mouse mesenchymal stem cells. Using fluorescence spectroscopy were investigated the interactions of new palladium(II) complexes [PdCl2(R2-S,S-pddtyr)]; (R = ethyl (C1), propyl (C2), butyl (C3), and pentyl (C4)) with calf thymus human serum albumin (HSA) and DNA (CT-DNA). The high values of the binding constants, Kb, and the Stern-Volmer quenching constant, KSV, show the good binding of all complexes for HSA and CT-DNA. The mentioned ligands and complexes were also tested on in vitro antimicrobial activity against 11 microorganisms. Testing was performed by the microdilution method, where the minimum inhibitory concentration (MMC) and the minimum microbicidal concentration (MMC) were determined.

A mini-review on indigenous microbial biofilm from various wastewater for heavy-metal removal - new trends.

Biofilm, as a form of the microbial community in nature, represents an evolutionary adaptation to the influence of various environmental conditions. In nature, the largest number of microorganisms occur in the form of multispecies biofilms. The ability of microorganisms to form a biofilm is one of the reasons for antibiotic resistance. The creation of biofilms resistant to various contaminants, on the other hand, improves the biological treatment process in wastewater treatment plants. Heavy metals cannot be degraded, but they can be transformed into non-reactive and less toxic forms. In this process, microorganisms are irreplaceable as they interact with the metals in a variety of ways. The environment polluted by heavy metals, such as wastewater, is also a source of undiscovered microbial diversity and specific microbial strains. Numerous studies show that biofilm is an irreplaceable strategy for heavy metal removal. In this review, we systematize recent findings regarding the bioremediation potential of biofilm-forming microbial species isolated from diverse wastewaters for heavy metal removal. In addition, we include some mechanisms of action, application possibilities, practical issues, and future prospects.

Synthesis, characterization, HSA binding, molecular docking, cytotoxicity study, and antimicrobial activity of new palladium(II) complexes with propylenediamine derivatives of phenylalanine.

The four new ligands, propylenediamine derivatives of phenylalanine (R2-S,S-pddbaˑ2HCl; L1-L4) and their palladium(II) complexes (C1-C4) were synthesized and characterized by elemental analysis, infrared, 1H and 13C NMR spectroscopy. The interactions of new palladium(II) complexes with human serum albumin (HSA) were studied by fluorescence spectroscopy. All investigated compounds can be transported to target cells by binding to HSA, but complex C4 interacts most strongly. Molecular docking simulations were applied to comprehend the binding of the complex to the molecular target of HSA. Obtained results are in good correlations with experimental data regarding binding affinity by HSA. In vitro cytotoxicity activities were investigated on four tumor cell lines (mouse mammary (4 T1) and colon (CT26), human mammary (MDA-MD-468) and colon (HCT116)) and mouse mesenchymal stem cells as non-tumor control cells. Cytotoxic capacity was determined by MTT test and according to obtained results ligand L4 stands out as the most active and selective compound and as a good candidate for future in vivo testing. Further examination of the ligand L4 and corresponding complex C4 led to the conclusion that both induced cell death mainly by apoptosis. Ligand L4 facilitated cycle arrest in G0/G1 phase and decreased proliferative capacity of tumor cells. In vitro antimicrobial activity for ligands and corresponding Pd(II) complexes was investigated against eleven microorganisms (eight strains of pathogenic bacteria and three yeast species) using microdilution method. The minimum inhibitory concentration and minimum microbicidal concentration were determined.

Putative application of Najas marina L. extracts as a source of bioactive compounds and their antioxidant, antimicrobial, antibiofilm, and genotoxic properties.

In this research paper, the total phenols (TP), flavonoids (TF), and tannins (TT) content in the acetone and ethyl acetate extracts of Najas marina L. and the identification and quantification of phenolic acids and flavonoids from the ethyl acetate extract were performed. Antioxidant, antimicrobial, and antibiofilm properties of the mentioned extracts were investigated in vitro. The genotoxic potential was analyzed in cultured human peripheral blood lymphocytes (PBL). The TP and TF content was higher in the ethyl acetate extract, dominated by quercetin (172.4 µg mg-1) and ferulic acid (22.74 µg mg-1), while the TT content was slightly higher in the acetone extract. Both extracts tested showed limited antioxidant effects compared to ascorbic acid. The strongest antibacterial activity was observed with Gram-positive bacteria, particularly Staphylococcus aureus (MIC and MMC at 0.31 mg ml-1) and S. aureus ATCC 25923 (MIC at <0.02 mg ml-1), while antifungal activity was limited. Both extracts tested showed better activity on preformed biofilms. Acetone extract had no genotoxic activity but showed significant genoprotective activity against mitomycin C-induced DNA damage in cultured PBLs. Results of our research demonstrate the potential for the development of plant-based antibacterial and biofilm agents.

Response of selected microbial strains and their consortia to the presence of automobile paints: Biofilm growth, matrix protein content and hydrolytic enzyme activity.

The goal of the current study was to examine the effects of pollutants (White color - CP; Metallic red color - FM; Thinner - CN; Thinner for rinsing paint - MF; Basic color (primer) - FH) originating from the automotive industry on the biofilm growth, matrix protein content, and activity of the hydrolytic enzymes of selected microbial strains in laboratory conditions that mimic the bioreactor conditions. The chosen microorganisms (bacteria, yeasts, and fungi) were isolated from automotive industry wastewater. Pure microbe cultures and their consortia were injected into AMB Media carriers and developed into biofilms. The use of AMB media carriers has been linked to an increase in the active surface area colonized by microorganisms. Afterwards, the carriers were transferred to Erlenmeyer flasks with nutrient media and pollutants at a concentration of 200 µL/mL. The current study found that, depending on the microbial strain, development phase, and chemical structure, the assessed pollutants had an inhibitory or stimulatory influence on the growth of single cultures and their consortia. Statistical analysis found positive correlations between the protein content in the matrix and the biofilm biomass of Rhodotorula mucilaginosa and consortia in CP and FH media, respectively. The proteolytic activity of Candida utilis was very pronounced in media with MF and CN. The best alkaline phosphatase activity (ALP) was achieved in the CN medium of R. mucilaginosa. Acid invertase activity was the highest in the FM and CP media of Escherichia coli and consortia, respectively, whereas the highest alkaline invertase activity was measured in the MF medium of E. coli. A positive correlation was confirmed between ALP and the biofilm biomass of R. mucilaginosa in CP and CN media, as well as between ALP and the biofilm biomass of Penicillium expansum in FM medium. The findings provide novel insights into the extracellular hydrolytic activity of the investigated microbial strains in the presence of auto paints, as well as a good platform for subsequent research into comprehensive biofilm profiling using modern methodologies.

Synthesis, characterization, DNA interactions and biological activity of new palladium(II) complexes with some derivatives of 2-aminothiazoles.

Newly palladium(II) complexes (C1, C2) with derivatives of 2-aminothiazoles (L1 = 2-amino-6-methylbenzothiazole, L2 = 2-amino-6-chlorobenzothiazole), general formula [PdL2Cl2] were synthesized and characterized by elemental microanalyses, IR, NMR spectroscopy and X-ray spectroscopy in case of [Pd(L2)2Cl2]. The kinetic of the substitution reactions of complexes and the nucleophiles, such as guanosine-5'-monophosphate (5'-GMP), tripeptide glutathione (GSH) and amino acid L-methionine (L-Met), were studied by stopped-flow technique. The complex C2 was always more reactive, while the order of the reactivity of the nucleophiles, due to the associative mode of the reaction, was L-Met > GSH > 5'-GMP. In order to determine the type of interactions between palladium(II) complexes and calf thymus DNA (CT-DNA), we used electronic absorption spectroscopy, viscosity measurements, and fluorescence spectroscopic studies, while interactions with bovine serum albumin (BSA) were determined only with fluorescence spectroscopic studies. The observed results confirmed that both complexes bound to DNA by groove binding. The significantly strong interaction with BSA, especially for complex C2, was also observed. In vitro cytotoxic activity was evaluated against four tumor cell lines, 4 T1, CT26, MDA-MB-468, HCT116 and mesenchymal stem cells (mMSC). C1 complex showed higher cytotoxic activity against CT26 cell line. Flow cytometry analysis showed that C1 stimulated apoptosis of tumor cells via inhibition of expression of antiapoptotic Bcl-2 molecule and decelerated proliferation by decreasing Cyclin-D and increasing expression of P21. In vitro antimicrobial activity for ligands and corresponding palladium(II) complexes was investigated by microdilution method and minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) were determined. Tested compounds exhibited selective and moderate activity.

Low-dimensional compounds containing bioactive ligands. Part XVII: Synthesis, structural, spectral and biological properties of hybrid organic-inorganic complexes based on [PdCl4]2- with derivatives of 8-hydroxyquinolinium.

In this study, four hybrid organic-inorganic compounds (8-H2Q)2[PdCl4] (1), (H2ClQ)2[PdCl4] (2), (H2NQ)2[PdCl4] (3) and (H2MeQ)2[PdCl4]·2H2O (4) (where 8-H2Q = 8-hydroxyquinolinium, H2ClQ = 5-chloro-8-hydroxyquinolinium, H2NQ = 5-nitro-8-hydroxyquinolinium and H2MeQ = 2-methyl-8-hydroxyquinolinium) were synthesized through organic cation modulation. Single-crystal X-ray structure analysis of compounds 1 and 3 indicates that their structures are planar and consist of [PdCl4]2- anions and 8-H2Q or H2NQ cations, respectively. Both ionic components are held together through ionic interactions and hydrogen bonds forming infinite chains linked through π-π interactions to form 2D structures. Furthermore, NMR spectroscopy, UV-Vis spectroscopy, elemental analysis, and FT-IR spectroscopy were used to explore the synthesized compounds. The DNA interaction, antimicrobial activity, antiproliferative activity, and radical scavenging effect of the compounds were evaluated. The hybrid compounds and their free ligands can interact with the calf thymus DNA via an intercalation mode involving the insertion of the aromatic chromophore between the base pairs of DNA; compound 1 has the highest binding affinity. Moreover, they have high antimicrobial efficacy against the tested 14 strains of microorganisms with minimum inhibitory concentration values ranging from <1.95 to 250 µg/mL. The antiproliferative activity of the compounds was investigated against three different cancer cell lines, and their selectivity was verified on mesenchymal stem cells. Compounds 1 and 2 displayed selective and high cytotoxicity against human lung and breast cancer cells and showed moderate cytotoxicity against colon cancer cells. Accordingly, they might be auspicious candidates for future pharmacological investigations in lung and breast cancer research.

Low-dimensional compounds containing bioactive ligands. Part VIII: DNA interaction, antimicrobial and antitumor activities of ionic 5,7-dihalo-8-quinolinolato palladium(II) complexes with K+ and Cs+ cations.

Starting from well-defined NH2(CH3)2[PdCl2(XQ)] complexes, coordination compounds of general formula Cat[PdCl2(XQ)] have been prepared by cationic exchange of NH2(CH3)2+ and Cat cations, where XQ are biologically active halogen derivatives of quinolin-8-ol (5-chloro-7-iodo-quinolin-8-ol (CQ), 5,7-dibromo-quinolin-8-ol (dBrQ) and 5,7-dichloro-quinolin-8-ol (dClQ)) and Cat is K+ or Cs+. The cation exchange of all prepared complexes, K[PdCl2(CQ)] (1), K[PdCl2(dClQ)] (2), K[PdCl2(dBrQ)] (3), Cs[PdCl2(CQ)] (4), Cs[PdCl2(dClQ)] (5) and Cs[PdCl2(dBrQ)] (6) was approved using IR spectroscopy, their structures in DMSO solution were elucidated by one- and two-dimensional NMR experiments, whereas their stability in solution was verified by UV-VIS spectroscopy. Interaction of complexes to ctDNA was investigated using UV-VIS and fluorescence emission spectroscopy. The minimum inhibitory concentration and the minimum microbicidal concentration values were detected against 15 bacterial strains and 4 yeast strains to examine the antimicrobial activity for the complexes. The in vitro antitumor properties of the complexes were studied by testing the complexes on leukemic cell line L1210, ovarian cancer cell line A2780 and non-cancerous cell line HEK293. The majority of the prepared compounds exhibited moderate antimicrobial and very high cytotoxic activity.

Low-dimensional compounds containing bioactive ligands. Part VI: Synthesis, structures, in vitro DNA binding, antimicrobial and anticancer properties of first row transition metal complexes with 5-chloro-quinolin-8-ol.

A series of new 3d metal complexes with 5-chloro-quinolin-8-ol (ClQ), [Mn(ClQ)2] (1), [Fe(ClQ)3] (2), [Co(ClQ)2(H2O)2] (3), [Ni(ClQ)2(H2O)2] (4), [Cu(ClQ)2] (5), [Zn(ClQ)2(H2O)2] (6), [Mn(ClQ)3]·DMF (7) and [Co(ClQ)3]·DMF·(EtOH)0.35 (8) (DMF=N,N-dimethylformamide), has been synthesized and characterized by elemental analysis, IR spectroscopy and TG-DTA thermal analysis. X-ray structure analysis of 7 and 8 revealed that these molecular complexes contain three chelate ClQ molecules coordinated to the central atoms in a deformed octahedral geometry and free space between the complex units is filled by solvated DMF and ethanol molecules. Antimicrobial activity of 1-6 was tested by determining the minimum inhibitory concentration and minimum microbicidal concentration against 12 strains of bacteria and 5 strains of fungi. The intensity of antimicrobial action varies depending on the group of microorganism and can be sorted: 1>ClQ>6>3/4>2>5. Complexes 1-6 exhibit high cytotoxic activity against MDA-MB, HCT-116 and A549 cancer cell lines. Among them, complex 2 is significantly more cytotoxic against MDA-MB cells than cisplatin at all tested concentrations and is not cytotoxic against control mesenchymal stem cells indicating that this complex seems to be a good candidate for future pharmacological evaluation. Interaction of 1-6 with DNA was investigated using UV-VIS spectroscopy, fluorescence spectroscopy and agarose gel electrophoresis. The binding studies indicate that 1-6 can interact with CT-DNA through intercalation; complex 2 has the highest binding affinity. Moreover, complexes 1-6 inhibit the catalytic activity of topoisomerase I.

ANTIMICROBIAL AND ANTIBIOFILM EFFECTS OF EXTRACTS FROM TRAPA NATANS L., EVALUATION OF TOTAL PHENOLIC AND FLAVONOID CONTENTS AND GC-MS ANALYSIS.

Research conducted in this study shows the applied in vitro antimicrobial and antibiofilm activity of the four extracts isolated from Trapa natans L. leaves. In this study, different methods were used (microdilution, tissue culture plate, different colorimetric methods, GC-FID and GC-MS analysis). While the water extract didn't show antibacterial activity, the acetone extract showed the strongest one. The same activity in the case of Pseudomonas aeruginosa (MIC was 313 µg/mL) was better than the activity of controls and it matched with antibiofilm activity. The effect of extracts was better on G+ bacteria (MICs were <78-625 µg/mL). For ethanol and ethyl acetate extracts all BIC values were better than MICs. Extracts showed a significant effect on Aspergillus restrictus (MICs were < 78/156 µg/mL). The GC and GC-MS analysis of the ethyl acetate extract revealed the identification of 22 compounds with (all E)-squalene (20.2%), n-alkanes and norlignan hinokiresinol among the most abundant ones. This is the first time that T. natans was studied using these methods.

Phytomedical investigation of Najas minor All. in the view of the chemical constituents.

Plants are an abundant natural source of effective antibiotic compounds. Phytomedical investigations of certain plants haven't still been conducted. One of them is Najas minor (N. minor), an aquatic plant with confirmed allelopathy. Research conducted in this study showed the influence of water and ethyl acetate extracts of N. minor on microorganisms, in the view of chemical profiling of volatile constituents and the concentrations of total phenols, flavonoids and tannins. Antimicrobial activity was defined by determining minimum inhibitory and minimum microbicidal concentrations using microdilution method. Influence on bacterial biofilm formation was performed by tissue culture plate method. The total phenolics, flavonoids and condensed tannins were determined by Folin-Ciocalteu, aluminum chloride and butanol-HCl colorimetric methods. Chemical profiling of volatile constituents was investigated by GC and GC-MS. Water extract didn't have antimicrobial activity below 5000 µg/mL. Ethyl acetate extract has shown strong antimicrobial activity on G+ bacteria - Staphylococcus aureus PMFKGB12 and Bacillus subtilis (MIC < 78.13 µg/mL). The best antibiofilm activity was obtained on Escherichia coli ATCC25922 (BIC50 at 719 µg/mL). Water extract had higher yield. Ethyl acetate extract had a significantly greater amount of total phenolics, flavonoids and tannins. As major constituent hexahydrofarnesyl acetone was identified. The ethyl acetate extract effected only G+ bacteria, but the biofilm formation of G-bacteria was suppressed. There was a connection between those in vivo and in vitro effects against pathogenic bacterial biofilm formation. All of this points to a so far unexplored potential of N. minor.

Biological evaluation of mechlorethamine-Pt(II) complex, part II: antimicrobial screening and lox study of the complex and its ligand.

The reaction of K(2)PtCl(4) with anticancer-alkylating agent mechlorethamine hydrochloride (CH(3)NH(C(2)H(4)Cl)(2) x HCl = HN2×HCl), in the molar ratio 1 : 2, affords the complex [H2N2](2)[PtCl(4)]. In vitro antimicrobial and lipoxygenase inhibitory activities of the complex and its precursor were evaluated. Antimicrobial activity of the HN2×HCl and [H2N2](2)[PtCl(4)] complex was investigated against 29 species of microorganisms. Testing is performed by microdilution method. Minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) have been determined. The difference between antimicrobial activity of precursor and corresponding platinum(II) complex is noticed and the activity of the precursor was higher. Tested compounds demonstrated the high and significant antifungal activity and low to moderate antibacterial activity. It was shown that the gram-positive bacteria were more sensitive than the gram-negative. UV absorbance-based enzyme assays were performed with HN2×HCl and [H2N2](2)[PtCl(4)] complex, in order to evaluate their in vitro inhibitory activity of soybean lipoxygenase (LOX), also. Assay with LOX showed significantly greater inhibitory activity of the complex, than the precursor.

Biological effects, total phenolic content and flavonoid concentrations of fragrant yellow onion (Allium flavum L.).

The antioxidant, antibacterial and antiproliferative activities, total phenolic content and concentrations of flavonoids of A. flavum extracts were determined. The total phenolic content was determined with Folin-Ciocalteu reagent and it ranged between 42.29 to 80.92 mg GA/g. The concentration of flavonoids in various extracts of A. flavum was determined using spectrophotometric method with aluminum chloride and obtained results varied from 64.07 to 95.71 mg RU/g. The antioxidant activity was monitored spectrophotometrically and expressed in terms of IC50 (µg/ml), and its values ranged from 64.34 to 243.34 µg/ml. The highest phenolic content and capacity to neutralize DPPH radicals were found in acetone extract. Antibacterial efficacy was defined by determining minimum inhibitory and minimum bactericidal concentrations using microdilution method. Significant antibacterial activity, especially for ethyl acetate extract, was observed. The best activity was showed against G+ bacteria, Staphylococcus aureus ATCC 25923 and Bacillus subtilis, while Escherichia coli was one of the least sensitive bacteria. Antiproliferative activity of the methanolic extract on HCT- 116 cell line was determined by MTT assay. Results showed that A. flavum has good antiproliferative activity with IC50 values of 28.29 for 24 h and 35.09 for 72 h. Based on these results, A. flavum is a potential source of phenols as natural antioxidant, antibacterial and anticancer substance of high value. Phenolic content of extracts depend on the solvents used for extraction.

Great horsetail (Equisetum telmateia Ehrh.): Active substances content and biological effects.

This paper deals with the antioxidant and antimicrobial activity, total phenolic content and concentrations of flavonoids of Equisetum telmateia extracts. Total phenolic content was determined with Folin-Ciocalteu reagent and it ranged between 129.0 to 262.7 mg GA/g. The concentration of flavonoids in various extracts of E. telmateia was determined using spectrophotometric method with aluminum chloride and obtained results varied from 112.6 to 199.8 mg RU/g. Antioxidant activity was monitored spectrophotometrically and expressed in terms of IC50 (µg/ml), and its values ranged from 33.4 to 982.2 µg/ml. The highest phenolic content, concentrations of flavonoids and capacity to neutralize DPPH radicals were found in the acetone extract. In vitro antimicrobial activity was determined using microdilution method. Minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) were also determined. Testing was performed on 22 microorganisms, including 15 strains of bacteria (standard and clinical strains) and 7 species of fungi. There were statistically significant differences in activity between the extracts of E. telmateia. Different effects were noticed against the bacteria and the methanol extract appeared to be most efficient. All the extracts showed significant antibacterial activity against G+ bacteria and weak to moderate activity against other microorganisms.

Biological activities of extracts from cultivated Granadilla Passiflora alata.

Research conducted in this study showed the influence of ethanol, acetone and ethyl acetate extracts of the outgrowth of cultivated Passiflora alata on microorganisms, as well as the antioxidant activity and the concentrations of total phenols, flavonoids and tannins. In vitro antimicrobial activities of extracts were studied on 27 species of microorganisms, of which 17 species of bacteria and 10 species of fungi. The strongest antimicrobial activity was detected on G+ bacteria while the activities on other species were moderate. Ethyl acetate extract showed the strongest effect. The concentrations of total phenols were examined by using Folin-Ciocalteu reagent and the obtained values ranged from 14.04 to 34.22 mg GA/g. By using aluminium chloride method, the concentrations of flavonoids were obtained and the values ranged from 33.19 to 62.30 mg RU/g. In determining the amount of tannins we used the method with buthanol-HCl reagent and the obtained value was 5.1 % of dry matter. The efficiency of antioxidation, which we identified through the reduction of DPPH, was in the range from 808.69 to 1107.79 µg/ml for a particular IC50, and AAI values were between 0.07 and 0.10. The best parameters were shown by ethanol extract. All data were statistically analyzed. Overall, extracts showed potential for further investigation and use.

Immortelle (Xeranthemum annuum L.) as a natural source of biologically active substances.

Antioxidant and antimicrobial effects, total phenolic content and flavonoid concentrations of methanolic, acetone and ethyl acetate extracts from Xeranthemum annuum L. were investigated in this study. The total phenolic content was determined using Folin-Ciocalteu reagent and ranged between 101.33 to 159.48 mg GA/g. The concentration of flavonoids in various X.annuum extracts was determined using spectrophotometric method with aluminum chloride and the results varied from 22.25 to 62.42 mg RU/g. Antioxidant activity was monitored spectrophotometrically using DPPH reagent and expressed in terms of IC50 (µg/ml), and it ranged from 59.25 to 956.81 µg/ml. The highest phenolic content and capacity to neutralize DPPH radicals were found in the acetone extract. In vitro antimicrobial activity was determined by microdilution method. Minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) have been determined. Testing was conducted against 24 microorganisms, including 15 strains of bacteria (standard and clinical strains) and 9 species of fungi. Statistically significant difference in activity between the extracts of X. annuum L. was observed and the acetone extract was found most active. The activity of acetone extract was in accordance with total phenol content and flavonoid concentration measured in this extract. The tested extracts showed significant antibacterial activity against G+ bacteria and weak to moderate activity against other microorganisms. Based on the obtained results, X. annuum can be considered as a rich natural source of polyphenolic compounds with very good antioxidant and antimicrobial activity.