s-Triazine Derivatives Functionalized with Alkylating 2-Chloroethylamine Fragments as Promising Antimicrobial Agents: Inhibition of Bacterial DNA Gyrases, Molecular Docking Studies, and Antibacterial and Antifungal Activity.

The spectrum of biological properties of s-triazine derivatives is broad and includes anti-microbial, anti-cancer, and anti-neurodegenerative activities, among others. The s-triazine molecule, due to the possibility of substituting three substituents, offers many opportunities to obtain hybrid compounds with a wide variety of activities. A group of 1,3,5 triazine derivatives containing a dipeptide, 2-ethylpiperazine, and a methoxy group as substituents was screened for their antimicrobial activity. An in vitro study was conducted on pathogenic bacteria (E. coli, S. aureus, B. subtilis, and M. luteus), yeasts (C. albicans), and filamentous fungi (A. fumigatus, A. flavus, F. solani, and P. citrinum) via microdilution in broth, and the results were compared with antibacterial (Streptomycin) and antifungal (Ketoconazole and Nystatin) antibiotics. Several s-triazine analogues have minimal inhibitory concentrations lower than the standard. To confirm the inhibitory potential of the most active compounds against gyrases E. coli and S. aureus, a bacterial gyrases inhibition assay, and molecular docking studies were performed. The most active s-triazine derivatives contained the -NH-Trp(Boc)-AlaOMe, -NH-Asp(OtBu)-AlaOMe, and -NH-PheOMe moieties in their structures.

Anti-multidrug-resistant Staphylococcus aureus and anti-dermatophyte activities of secondary metabolites of the endophytic fungus Penicillium brevicompactum ANT13 associated with the Algerian endemic plant Abies numidica.

This study aims to identify and assess the antimicrobial activity of endophytic fungi found in the endemic plant Abies numidica. Among all isolates, the ANT13 isolate demonstrated significant antimicrobial activity in the preliminary screening, particularly Staphylococcus aureus ATCC 25923 and Candida albicans ATCC 1024, with inhibition zones of 22 and 21.5 mm, respectively. Based on its morphological and molecular features, this isolate was identified as Penicillium brevicompactum. The maximum activity was observed in the ethyl acetate extract, followed by the dichloromethane extract; however, the n-hexane extract exhibited no activity. The ethyl acetate extract demonstrated very significant activity against the five strains of multidrug-resistant Staphylococcus aureus used, with average zones of inhibition ranging from 21 to 26 mm, in contrast to more resistant Enterococcus faecalis ATCC 49452 and Bacillus cereus ATCC 10876. The ethyl acetate extract was also very active against dermatophytes, where the zones of inhibition for Candida albicans, Microsporum canis, Trichophyton mentagrophytes, Trichophyton rubrum, and Epidermophyton floccosum were 23.5, 31, 43, 47, and 53.5 mm, respectively. The MIC values for dermatophytes ranged between 100 and 3200 µg/mL. The wild isolate of Penicillium brevicompactum ANT13 discovered as an endophyte in Abies numidica may be a distinctive source of novel compounds and drug discovery to trait dermatophytes and multidrug-resistant Staphylococcus aureus infections.

Design, Synthesis, Anti-Proliferative, Anti-microbial, Anti-Angiogenic Activity and In Silico Analysis of Novel Hydrazone Derivatives.

BACKGROUND: Cancer is the second leading cause of death globally. Hydrazone and hydrazone derivatives have high activity, and for this reason, these compound are greatly used by researchers to synthesize new anti-cancer drug. The aim of this research work is to synthesize novel anticancer agents. METHODS: New hydrazone derivatives were synthesized via a reaction between 3-formylphenyl methyl carbonate and benzhydrazide, 4-methylbenzoic hydrazide, 4-tert-butylbenzoic hydrazide, 4-nitrobenzoic hydrazide and 3- methoxybenzoic hydrazide, and were successfully characterized using elemental analysis, 1H-NMR, 13C-NMR, FT-IR and LC-MS techniques. The synthesized compounds were evaluated for their antimicrobial (some grampositive and -negative bacteria, filamentous fungi and yeasts), anti-proliferative (T47D and HCC1428-breast cancer cells) and anti-angiogenic (HUVEC-endothelial cells) activities. The anti-proliferative activities of the hydrazone compounds R1-R5 were studied on these cell lines by MTT assay. The anti-angiogenic potential of the compounds was determined by the endothelial tube formation assay. To identify structural features related to the anti-proliferative activity of these compounds, 2D-QSAR was performed. RESULT: The results indicated that compound R3 exhibited strong anti-angiogenic and anti-proliferative activity on breast cancer cell lines and healthy cell lines. Also, this compound; possessing a tertiary butyl moiety on the hydrazine, exhibited the highest inhibitory effect against all tested microorganisms; in particular, it inhibited Candida albicans at a lower concentration than ketoconazole. Among the investigated compounds, those bearing methyl, tertiary butyl (compound R2, R3) and methoxy (compound R5) moiety were found to be more successful anticandidal derivatives than standard antifungal antibiotics. The QSAR analysis suggested that the tumor specificity of the hydrazone correlated with their molecular weight, lipophilicity, molar refractivity, water solubility, DipolHybrid:(MOPAC) and ExchangeEnergy:(MOPAC). Absorption, Distribution, Metabolism and Elimination (ADME) analysis of the hydrazone compounds showed that they have favorable pharmacokinetic and drug-likeness properties. The ADME results clarify that R3 is the best compound in terms of pharmacokinetic properties. In contrast to other compounds; target prediction analysis of the compound R3 showed inhibitory activity on estrogen-related receptor alpha transcription factor (ESRRA). The target prediction analysis was supported by molinspiration bioactivity score. CONCLUSION: The R3 compound is considered to be an important candidate for future studies with its suitability for the Lipinski's rule of five for drug-likeness, and effective in vitro and in silico results.

Fungal contaminants in man-made water systems connected to municipal water.

Water-related fungi are known to cause taste and odor problems, as well as negative health effects, and can lead to water-pipeline clogging. There is no legal regulation on the occurrence of fungi in water environments. However, much research has been performed, but further studies are needed. The main objectives of this study were to evaluate the fungal load and the presence of mycotoxigenic fungi in man-made water systems (for homes, hospitals, and shopping centers) connected to municipal water in Istanbul, Turkey. The mean fungal concentrations found in the different water samples were 98 colony-forming units (CFU)/100 mL in shopping centers, 51 CFU/100 mL in hospitals, and 23 CFU/100 mL in homes. The dominant fungal species were identified as Aureobasidium pullulans and Fusarium oxysporum. Aflatoxigenic Aspergillus flavus and ochratoxigenic Aspergillus westerdijkiae were only detected in the hospital water samples. Alternaria alternata, Aspergillus clavatus, Aspergillus fumigatus, and Cladosporium cladosporioides were also detected in the samples. The study reveals that the municipal water supplies, available for different purposes, could thus contain mycotoxigenic fungi. It was concluded that current disinfection procedures may be insufficient, and the presence of the above-mentioned fungi is important for people with suppressed immune systems.

Indoor airborne fungal pollution in newborn units in Turkey.

Pathogenic and/or opportunistic fungal species are major causes of nosocomial infections, especially in controlled environments where immunocompromised patients are hospitalized. Indoor fungal contamination in hospital air is associated with a wide range of adverse health effects. Regular determination of fungal spore counts in controlled hospital environments may help reduce the risk of fungal infections. Because infants have inchoate immune systems, they are given immunocompromised patient status. The aim of the present study was to evaluate culturable airborne fungi in the air of hospital newborn units in the Thrace, Marmara, Aegean, and Central Anatolia regions of Turkey. A total of 108 air samples were collected seasonally from newborn units in July 2012, October 2012, January 2013, and April 2013 by using an air sampler and dichloran 18% glycerol agar (DG18) as isolation media. We obtained 2593 fungal colonies comprising 370 fungal isolates representing 109 species of 28 genera, which were identified through multi-loci gene sequencing. Penicillium, Aspergillus, Cladosporium, Talaromyces, and Alternaria were the most abundant genera identified (35.14, 25.40, 17.57, 2.70, and 6.22% of the total, respectively).

Fungal biodiversity and mycotoxigenic fungi in cooling-tower water systems in Istanbul, Turkey.

This is the first study to assess fungal diversity and mycotoxigenic fungi in open recirculating cooling-tower (CT) water systems (biofilm and water phase). The production capability of mycotoxin from fungal isolates was also examined. The mean fungal count in 21 different water and biofilm samples was determined as 234 CFU/100 mL and 4 CFU/cm2. A total of 32 species were identified by internal transcribed spacer (ITS) sequencing. The most common isolated fungi belonged to the genera Aspergillus and Penicillium, of which the most prevalent fungi were Aspergillus versicolor, Aspergillus niger, and Penicillium dipodomyicola. From 42% of the surveyed CTs, aflatoxigenic A. flavus isolates were identified. The detection of opportunistic pathogens and/or allergen species suggests that open recirculating CTs are a possible source of fungal infection for both the public and for occupational workers via the inhalation of aerosols and/or skin contact.

Synthesis and biological evaluation of new naphthalene substituted thiosemicarbazone derivatives as potent antifungal and anticancer agents.

New thiosemicarbazone derivatives (1-10) were obtained via the reaction of 4-(naphthalen-1-yl)thiosemicarbazide with fluoro-substituted aromatic aldehydes. The synthesized compounds were evaluated for their in vitro antifungal effects against pathogenic yeasts and molds using broth microdilution assay. Ames and umuC assays were carried out to determine the genotoxicity of the most effective antifungal derivatives. Furthermore, all compounds were evaluated for their cytotoxic effects on A549 human lung adenocarcinoma and NIH/3T3 mouse embryonic fibroblast cell lines using XTT test. Among these derivatives, 4-(naphthalen-1-yl)-1-(2,3-difluorobenzylidene)thiosemicarbazide (1) and 4-(naphthalen-1-yl)-1-(2,5-difluorobenzylidene)thiosemicarbazide (3) can be identified as the most promising antifungal derivatives due to their notable inhibitory effects on Candida species and no cytotoxicity against NIH/3T3 mouse embryonic fibroblast cell line. According to Ames and umuC assays, compounds 1 and 3 were classified as non-mutagenic compounds. On the other hand, 4-(naphthalen-1-yl)-1-(2,4-difluorobenzylidene)thiosemicarbazide (2) can be considered as the most promising anticancer agent against A549 cell line owing to its notable inhibitory effect on A549 cells with an IC50 value of 31.25 µg/mL when compared with cisplatin (IC50 = 16.28 µg/mL) and no cytotoxicity against NIH/3T3 cells.

A novel series of thiazolyl-pyrazoline derivatives: synthesis and evaluation of antifungal activity, cytotoxicity and genotoxicity.

In the current work, new thiazolyl-pyrazoline derivatives (1-22) were synthesized and evaluated for their antifungal effects against pathogenic yeasts and molds using a broth microdilution assay. Ames assay was carried out to determine the genotoxicity of the most effective antifungal derivatives. The cytotoxicity of the compounds (1-22) was also investigated against A549 human lung adenocarcinoma and NIH/3T3 mouse embryonic fibroblast cells. Among these derivatives, 2-[5-(4-fluorophenyl)-3-(5-methylthiophen-2-yl)-4,5-dihydro-1H-pyrazol-1-yl]-4-(4-methylsulfonylphenyl)thiazole (18) can be identified as the most promising anticandidal derivative due to its notable inhibitory effect on Candida zeylanoides with a MIC value of 250 µg/mL when compared with ketoconazole (MIC = 250 µg/mL), low cytotoxicity against NIH/3T3 cells and non-mutagenic effect. On the other hand, 2-[5-(4-fluorophenyl)-3-(5-chlorothiophen-2-yl)-4,5-dihydro-1H-pyrazol-1-yl]-4-(4-bromophenyl)thiazole (4) can be considered as the most promising anticancer agent against A549 cancer cells owing to its notable inhibitory effect on A549 cells with an IC50 value of 62.5 µg/mL when compared with cisplatin (IC50 = 45.88 µg/mL) and low cytotoxicity against NIH/3T3 cells.

Synthesis and biological evaluation of thiazoline derivatives as new antimicrobial and anticancer agents.

N'-(3,4-Diarylthiazol-2(3H)-ylidene)-2-(arylthio)acetohydrazides were synthesized and evaluated for their antimicrobial activity and cytotoxicity against NIH/3T3 cells. Compound 22 bearing 1-phenyl-1H-tetrazole and p-chlorophenyl moieties was found to be the most promising antibacterial agent against Pseudomonas aeruginosa, whereas compound 23 bearing 1-phenyl-1H-tetrazole and p-bromophenyl moieties was the most promising antifungal agent against Candida albicans. The most effective derivatives were also evaluated for their cytotoxicity against C6 glioma cells. The results indicated that compound 17 bearing 1-phenyl-1H-tetrazole and nonsubstituted phenyl moieties (IC50 = 8.3 ± 2.6 µg/mL) was more effective than cisplatin (IC50 = 13.7 ± 1.2 µg/mL) against C6 glioma cells. Compound 17 also exhibited DNA synthesis inhibitory activity on C6 cells. Furthermore, compound 17 showed low toxicity to NIH/3T3 cells (IC50 = 416.7 ± 28.9 µg/mL).

Mycotoxigenic moulds and mycotoxins in flours consumed in Turkey.

BACKGROUND: Aflatoxins (AFs) and ochratoxin A (OTA) are metabolites produced by several fungi of the genera Aspergillus and Penicillium and have been found to contaminate human foods and animal feeds. The aim of this study was to investigate the abundance and diversity of total microfungi and mycotoxigenic fungi in 25 samples of different grain-based flours from four regions of Turkey (Thrace and Central, Northwest and West Anatolia) and to evaluate the level of AF and OTA contamination. Microscopic and polymerase chain reaction analyses were used to identify fungi, while high-performance liquid chromatography was used for the detection of AFs and OTA. RESULTS: A total of 551 fungal strains were obtained from the samples and identified morphologically and by multi-locus gene sequencing. All samples were contaminated with fungi ((2-4.8) × 10(4) colony-forming units g(-1) ) and three of them exceeded the European Commission (EC) limits. The data also revealed that 70.5 and 14.7% of the fungal isolates were positive for AF and OTA production respectively. In addition, 21 samples were contaminated by AFs (14.98 and 22.4 µg kg(-1) for AFB1 ) and OTA (3.02 and 4.76 µg kg(-1) ) and three of them exceeded the EC limits. CONCLUSION: This study is the first report on problems with the occurrence of microfungi, mycotoxin contamination, AFs and OTA in different grain-based flour samples from Turkey and highlights developable points of current limits for food and public health safety.

Polymerase chain reaction (PCR) identification of terverticillate Penicillium species isolated from agricultural soils in eskişehir province

In the present study, nine terverticillate Penicillium isolates (P. griseofulfum, P. puberulum, P. crustosum, P. aurantiogriseum, P. chrysogenum, P. primulinum, P. expansum, P. viridicatum, Eupenicillium egyptiacum) from 56 soil samples were characterized genetically by a PCR method. The DNAs of the strains were isolated using the glass beads and vortexing extraction method and then used for PCR amplification with the internal transcribed spacer 1 (ITS1) and ITS4 universal fungal specific primers. The ITS regions of fungal ribosomal DNA (rDNA) were sequenced through the CEQ 8000 Genetic Analysis System. ITS-5.8S sequences obtained were compared with those deposited in the GenBank Database. The results indicated that the identification of Penicillium species with PCR based methods provided significant information about the solution to taxonomy and improve food safety and to protect the users from harmful contaminants such as mycotoxins, which must be controlled during the production of agricultural materials as well as during the processing of food and feed.

Separation and purification of hyaluronic acid by embedded glucuronic acid imprinted polymers into cryogel.

Hyaluronic acid (HA) has been used in many applications such as pharmaceutical, clinical and cosmetics, so its separation and purification is very important. In this study, firstly d-glucuronic acid imprinted polymers (MIPs) have been synthesized for the separation of HA which has glucuronic acid part in its structure. MIP particles have characterized by elemental analysis, Fourier Transform Infrared Spectroscopy (FT-IR) and swelling tests. Then, synthesized MIP particles have embedded into polyacrylamide based cryogel. Cryogel has prepared by free radical cryogelation process initiated by N,N,N',N'-Tetramethylethylenediamine (TEMED) and ammonium persulfate (APS) as redox initiators. This cryogel material was characterized by FT-IR, swelling tests, scanning electron microscopy (SEM) and surface adsorption analyze including pore size analyzer (BET) method. The adsorption of HA has investigated by spectrophotometric method using MIPs embedded into cryogel columns (GAIPEC) and the maximum HA adsorption capacity was found to be 318mgg(-1). The selectivity of GAIPEC column has estimated using N-acetylglucose amine as interfering agent since this molecule is a part of HA and the results have shown that GAIPEC has been nearly 35 times selective for HA than N-acetylglucose amine. The optimum chromatographic conditions for separation of HA were investigated. pH 7.0 buffer solution for elution and 0.1M of NaCl solution as desorption agent were used at 0.5mLmin(-1) flow rate. Also, recovery of GAIPEC was investigated and the results have shown that GAIPEC could be used many times without decreasing its adsorption capacity significantly. Here in, combining selectivity of MIP particles and mechanical properties of cryogel, a rigid and stable material was prepared for the separation and purification of HA. To point out this, HA has been isolated from fish eye and fermentation of Streptococcus equi RSKK 679 cell culture. After that, it has characterized and Fast Protein Liquid Chromatography (FPLC) applications have been investigated.

Microfungi in cultivated fields in Eskisehir provience (Turkey).

The soil microfungi flora was investigated in four locations of Eskisehir (Turkey). 56 soil samples were seasonaly collected from 14 stations in the areas of Karacahöyük, Bahçecik, OGU I, and OGU II. A total of 110 species belonging to 32 genera were encountered including Absidia, Acremonium, Alternaria, Aspergillus, Beauveria, Botryoderma, Chaetomium, Chrysosporium, Cladosporium, Eupenicillium, Eurotium, Fusarium, Geotrichum, Gliocladium, Gonytrichum, Metarrhizium, Mucor, Myrothecium, Paecilomyces, Penicillium, Phoma, Plectosphaerella, Rhizoctania, Rhizopus, Scopulariopsis, Septonema, Stachybotrys, Trichocladium, Trichoderma, Ulocladium, Verticillium, and Wardomyces. Twenty five species were more frequent (all locations) while twenty seven species were rare (only one sample). Mainly, Acremonium kiliense, Aspergillus ochraceus, A. terricola var. americanus, A. versicolor, Cladosporium cladosporioides, Fusarium oxysporum, F. solani, Gliocladium roseum, Penicillium chrysogenum, P. corylophum, P. expansum, P. griseofulvum, P. implicatum, P. restrictum, and Stachybotrys chartarum were the most common and abundant microfungi in all locations. Five species Aspergillus subsessilis, A. terreus var. africanus, Eupenicillium egyptiacum, Paecilomyces ramosus, and Penicillium novae-zeelandiae are likely to be newly recorded for Turkey. The microfungi number in Eskisehir soils was between 25,000-234,000 CFU/g (mean value at 126,375 CFU/g).