[Effect of Phenylalanine-arginine-beta-naphthylamide to Ciprofloxacin Minimum Inhibitory Concentration Values and Expression of Efflux Pump System Genes in Acinetobacter baumannii Isolates]. / Acinetobacter baumannii Izolatlarinda Fenilalanin-arjinin-beta-naftilamidin Siprofloksasinin Minimal Inhibitör Konsantrasyon Degerleri ve Disa Atim Pompasi Genlerinin Ekspresyonu Üzerine Etkisi.

The most realistic approach in recent years is researching the resistance inhibition rather than synthesizing new compounds. In this study, we aimed to determine i) the effect of phenylalanine-argininebeta-naphthylamide (PAßN), on minimum inhibition concentrations (MICs) of ciprofloxacin (CIP), ii) to obtain the CIP+PAßN concentration that inhibits CIP resistance and iii) to show that this inhibition is caused by the effect of PAßN on the expression of efflux pump (EF) system genes. Acinetobacter baumannii isolates were collected from Trakya University Hospital. In 67 isolates determined to be resistant to CIP, CIP susceptibility was investigated in presence of PAßN once again. Isolates determined to have four or more fold decrease in ciprofloxacin MIC values were included in checkerboard assay and quantitative real-time reverse transcriptase polymerase chain reaction (qrRT-PCR). Fractional inhibition concentrations (FICs) were calculated through the PAßN concentrations that inhibit ciprofloxacin resistance, by the checkerboard assay results. With the combination of CIP+PAßN, the effect of the concentrations at which inhibition occurs, to the expression levels of EF system genes (adeA, adeB, adeR, adeS, adeF, adeG, adeH, adeL) was investigated by qrRT-PCR. By the checker board assay, a synergistic effect was determined between PAßN and CIP in 11 isolates, while in other isolates the effect was determined to be additive. In some isolates resistant to CIP, CIP + PAßN combination inhibited the resistance and increased CIP susceptibility. In the presence of 25 mg/L and 100 mg/L PAßN, 22 (32.83%) and 27 (40.3%) of 67 isolates became sensitive to CIP, respectively. In seven isolates, 12.5 µg/ml PAßN concentration eliminated CIP resistance by decreasing CIP MIC value to 1 µg/ml. Also, in one isolate the MIC value was 0.5 µg/ml in the presence of 25 µg/ml PAßN and 1 µg/ml in the presence of 1.5625 µg/ml PAßN. After analyzing the expression levels of EF genes (adeA, adeB, adeC, adeF, adeG, adeH, adeL, adeR and adeS) by the qRt-PCR method, it was determined that with the addition of PAßN to media containing CIP, the expression levels of the genes decreased (p<0.05). The aim of the study has been achieved with the results obtained. These results highlighted the importance of research on the inhibition of resistance, as well as the synthesis of new antimicrobial compounds. Combined use of inhibitors and antibiotics should be considered as an alternative treatment method. Thus, existing antibiotics can be included in the treatment again, saving time and money. It will be possible to use these findings in further studies to elucidate the mechanism of action of new inhibitor candidate compounds and associate them with the expression of DAP genes, also by investigating mutations in the regulatory gene regions in isolates with over-expression levels.

Synthesis and molecular docking studies of some novel antimicrobial benzamides.

Common use of classical antibiotics has caused to the growing emergence of many resistant strains of pathogenic bacteria. Therefore, we aimed to synthesize a number of N-(2-hydroxy-(4 or 5)-nitrophenyl)benzamide derivatives as a new class of antimicrobial compounds. Moreover, our second goal is to predict the interaction between active structures and enzymes (DNA -gyrase and FtsA) in the binding mode. In this study, thirteen N-(2-hydroxy-(4 or 5-nitrophenyl)-substituted-benzamides were synthesized and determined for their antimicrobial activity using the microdilution method. According to this work, none of the compounds showed any activity against Candida albicans and its clinical isolate. Some of the benzamides (4N1, 5N1, 5N2) displayed very significant activity against Staphylococcus aureus and MSSA with <4 µg/ml MIC value, even they were found to be more potent than ceftazidime. 4N1 was also found to be more effective than gentamicin against Enterococcus faecalis clinical isolate. Molecular docking studies revealed that 4N1, 5N1, and 5N2 showed a good interactions with DNA-gyrase. Moreover, 5N1 has interacted with FtsA enzyme in the binding mode, as well. Only compound 5N4 displayed very good activity against Escherichia coli ATCC 25922. These findings showed us that 4N1, 5N1, 5N2, and 5N4 could be lead compounds to discover new antibacterial candidates against multidrug-resistant strains.

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).

Synthesis and antimicrobial evaluation of some novel sulfonylamido-benzoxazoles.

A series of 2-(p-substituted phenyl)-5-[(4-substituted phenyl) sulfonylamido]-benzoxazoles were synthesized and tested for their antimicrobial activities. The structures of the new derivatives were elucidated by spectral techniques. The minimum inhibitory concentrations (MIC) of the new benzoxazoles were determined against standard bacterial and fungal strains and drug-resistant isolates and compared to those of several reference drugs.

Aspergillus alliaceus, a new potential biological control of the root parasitic weed Orobanche.

During extensive surveys in fields heavily infested by broomrape in the Trakya Region-Turkey, a different new fungus, Aspergillus alliaceus, was isolated from the infected broomrape. It is aimed to investigate whether or not it is really a pathogen for Orobanche. The fungi was exposed to a greenhouse environment in order to assess its pathogenicity and virulence against Orobanche cernua. In addition, infection tests on Orobanche seeds were also performed under laboratory conditions. The fungus was subjected using two different methods, exposure to a liquid culture with conidial solution and a sclerotial solid culture with fungal mycelia. Cytological studies were carried out at light, TEM and SEM levels. The results show that the sclerotial solid culture with fungal mycelia quickly caused necrosis and was more effective than the other type. It also greatly diminished attachments, tubercles, and caused the emergence of shoots and an increase in the total shoot number of Orobanche. In addition, both when the fungi was exposed to both soil and used to contaminate sunflower seeds, its pathogenicity was more effective. Consequently, it was determined that A. alliaceus was an effective potential biological control of broomrape throughout its life cycle from dormant seed to mature plant.

Airborne fungi and bacteria in indoor and outdoor environment of the Pediatric Unit of Edirne Government Hospital.

This study was performed between January 2004 and December 2004 in 13 stations in the Pediatric Unit of Edirne Government Hospital in order to determine the outdoor and indoor airborne microfungal and bacterial contents. The results of air samplings revealed that 1,376 microfungal and 2,429 bacterial colonies in total were isolated. The isolated microfungal specimens were identified and 65 species from 16 genera were determined. Among these, the most frequent genus was Cladosporium with 462 colonies (33.58%) followed by Alternaria with 310 (22.53%) and Penicillium with 280 (20.35%) colonies. The isolated bacterial samples were grouped based on their Gram-staining properties. The most frequent ones were Gram (+) cocci with 1,527 colonies (62.87%) followed by Gram (+) bacilli with 828 colonies (34.09%) and Gram (-) bacilli with 74 colonies (3.05%). Staphylococcus, Bacillus, Corynebacterium, and Microccus appeared to be the common genera isolated for all months. Statistical analyses were performed in order to see if there existed a relationship between meteorological conditions and the microfungal and bacterial species and their concentrations.

A novel biosensor based on Lactobacillus acidophilus for determination of phenolic compounds in milk products and wastewater.

Different branches of industry need to use phenolic compounds (PCs) in their production, so determination of PCs sensitively, accurately, rapidly, and economically is very important. For the sensitive determination of PCs, some biosensors based on pure polyphenol oxidase, plant tissue and microorganisms were developed before. But there has been no study to develop a microbial phenolic compounds biosensor based on Lactobacillus species, which contain polyphenol oxidase enzyme. In this study, we used different forms of Lactobacillus species as enzyme sources of biosensor and compared biosensor performances of these forms for determination of PCs. For this purpose, we used lyophilized Lactobacillus cells (containing L. bulgaricus, L. acidophilus, Streptococcus thermophilus), pure L. acidophilus, pure L. bulgaricus, and L. acidophilus- and L. bulgaricus adapted to catechol in Lactobacilli MRS Broth. The most suitable form was determined and optimization studies of the biosensor were carried out by using this form. For preparing the bioactive layer of the biosensor, the Lactobacillus cells were immobilized in gelatin by using glutaraldehyde. In the study, we used catechol as a substrate. Phenolic compound determination is based on the assay of the differences on the respiration activity of the cells on the oxygen meter in the absence and the presence of catechol. The microbial biosensor response depends directly on catechol concentration between 0.5 and 5.0 mM with 18 min response time. In the optimization studies of the microbial biosensor the most suitable microorganism amount was found to be 10 mg, and also phosphate buffer (pH 8.0; 50 mM) and 37.5 °C were obtained as the optimum working conditions. In the characterization studies of the microbial biosensor some parameters such as substrate specificity on the biosensor response and operational and storage stability were examine. Furthermore, the determination of PC levels in synthetic wastewater, industrial wastewater, and milk products was investigated by using the developed biosensor under optimum conditions.

Investigation of indoor molds and allergic diseases in public primary schools in Edirne city of Turkey.

BACKGROUND: Studies evaluating the role of indoor molds in the development of allergic or respiratory symptoms in schools are few in childhood. OBJECTIVE: This study aimed to investigate relation between indoor molds and allergic diseases or respiratory symptoms in primary school's children in Edirne, Turkey. METHODS: Ten public primary schools were included into the study. A thorough assessment, using a questionnaire and inspection surveys was carried out. The concentration of culturable mold was assessed in the dust samples in the schools. Indoor temperature and humidity were measured. A total of 1374 students who completed valid questionnaires were included in the study, and dust-samples were collected from the schools. RESULTS: Cumulative and current prevalence rates of wheezing, asthma, allergic rhinitis, and atopic dermatitis were found as 31.4%, 9.3%, 16.2%, 6.0% and 13.4%, 11.9%, 15.1%, 2.1%, respectively. The most frequent mold-species detected in indoor dusts were Cladosporium, Penicillium, Alternaria and Aspergillus. Although the prevalence rates of allergic diseases and respiratory symptoms were high, indoor mold amounts were low in the schools in our region and no significant correlation was determined between indoor mold amount and the prevalence of these diseases in schools or classrooms. CONCLUSION: Even though allergic molds are present in schools, the mold-exposure may not be an important predisposing factor for development of allergic and respiratory diseases the schools in our region.

Design, synthesis, molecular docking, density functional theory and antimicrobial studies of some novel benzoxazole derivatives as structural bioisosteres of nucleotides.

A series of some novel 2-(p-tert-butylphenyl)-5-(3-substituted-propionamido)benzoxazole derivatives have been designed, synthesized, evaluated for antimicrobial activity and have performed molecular docking studies against penicillin-binding protein 4 (PBP4) and active and allosteric site of PBP2a; were calculated some theoretical quantum parameters and absorption, distribution, metabolism and excretion (ADME) descriptors. B9 acted at minimum inhibitory concentration (MIC) = 8 µg/mL against S. aureus, E. faecalis and their drug-resistant isolates and also formed with GLU145 (1.74 Å) and ILE144 (1.89 Å) two hydrogen bonds at allosteric site of PBP2a with Glide emodel score: -42.168. ΔE of compound B9 had moderate value of all compounds with 0.14742.[Formula: see text]Communicated by Ramaswamy H. Sarma.

Airborne and soilborne microfungi in the vicinity Hamitabat Thermic Power Plant in Kirklareli City (Turkey), their seasonal distributions and relations with climatological factors.

Soil and air samples of seven different localities around Hamitabat Thermic Power Plant, 10 km far away from Luleburgaz/Kirklareli (Turkey), were taken between the years 2003 and 2004 with seasonal intervals. The samples were brought to the laboratory and their microfungal identifications were done. From the air samples, 737 microfungi colonies were isolated comprising 26 species belonging to eight genera. From soil samples, 170.6 x 10(4) colony-forming unit (CFU)/g was isolated from 33 species belonging to 16 genera. The most isolated genus from air samples was Alternaria (324 CFU, 43.96%), followed by Cladosporium (208 CFU, 25.52%) and Phoma (44 CFU, 5.40%). Penicillium was the most isolated genus from the soil samples with a value of 560,000 CFU/g (32.8%), followed by Fusarium (226,000 CFU/g, 13.12%) and Aspergillus (154,000 CFU/g, 9.03%). Among these species, Alternaria citri and Alternaria alternata are the most abundant species in air with 164 and 107 CFU, respectively, whereas Fusarium graminearum and Penicillium citrinum are the most abundant species in soil with CFU per gram values of 17.8 x 10(4) and 1.3 x 10(5). Correlation analysis was applied to determine whether or not there was a relationship between colony number of isolated fungal genera and meteorological factors. Some parameters of soil samples' incontent during the research period were calculated using a computer analysis program. From the air samples, a positive correlation was found between relative humidity and Alternaria colonial counts and Cladosporium spore counts (r = 0.912 and r = 1.000, respectively). Similarly, with the analysis of soil samples, a positive correlation between colonial counts of Alternaria and soil pH and a positive correlation between colonial counts of Aspergillus and Penicillium and salt percentage concentration of soil were found.

Mycological contamination in dental unit waterlines in Istanbul, Turkey.

Studies on dental units (DUs) are conducted either for the prevention or the reduction of the density of bacterial contamination in dental unit waterlines (DUWLs). However, the existence of fungi in the these systems requires more attention. During dental treatment, direct contact with water contaminated with fungi such as Candida, Aspergillus, or inhalation of aerosols from high-speed drill may cause various respiratory infections, such as asthma, allergies, and wounds on mucose membranes, especially on immunocompromised patients and dentists. The aims of this study are to investigate the number and colonization of fungi in DUWLs in the city of Istanbul, Turkey. Water samples were collected from air-water syringes, high-speed drills, and inlet waters from 41 DUs. The aerobic mesophilic fungi count in high- speed drills was higher than inlet waters and air-water syringes. Non-sporulating fungi were found in 7 DUs. The isolated fungi were identified as Penicillium waksmanii, Cladosporium spp., Penicillium spp., Candida famata, Cryptococcus laurentii, Candida guilliermondii, Penicillium verrucosum, Aspergillus pseudoglaucus, Penicillium decumbens, and Acremonium sp. Some of these fungal genera are known as opportunistic pathogens that led to respiratory diseases such as allergic rhinits. This study shows the importance of regular control of mycological contamination on water at DUs.

Mycological contamination in dental unit waterlines in Istanbul, Turkey

Studies on dental units (DUs) are conducted either for the prevention or the reduction of the density of bacterial contamination in dental unit waterlines (DUWLs). However, the existence of fungi in the these systems requires more attention. During dental treatment, direct contact with water contaminated with fungi such as Candida, Aspergillus, or inhalation of aerosols from high-speed drill may cause various respiratory infections, such as asthma, allergies, and wounds on mucose membranes, especially on immunocompromised patients and dentists. The aims of this study are to investigate the number and colonization of fungi in DUWLs in the city of Istanbul, Turkey. Water samples were collected from air-water syringes, high-speed drills, and inlet waters from 41 DUs. The aerobic mesophilic fungi count in highspeed drills was higher than inlet waters and air-water syringes. Non-sporulating fungi were found in 7 DUs. The isolated fungi were identified as Penicillium waksmanii, Cladosporium spp., Penicillium spp., Candida famata, Cryptococcus laurentii, Candida guilliermondii, Penicillium verrucosum, Aspergillus pseudoglaucus, Penicillium decumbens, and Acremonium sp. Some of these fungal genera are known as opportunistic pathogens that led to respiratory diseases such as allergic rhinits. This study shows the importance of regular control of mycological contamination on water at DUs.

Airborne Alternaria and Cladosporium species and relationship with meteorological conditions in Eskisehir City, Turkey.

Alternaria and Cladosporium, known as the most allergenic spores were first collected by means of Durham gravimetric sampler from Eskisehir atmosphere from January 1, 2000 to December 31, 2001. The daily, monthly and annual variations in spores/cm(2) of Cladosporium and Alternaria were recorded. During this period, a total of 10.231 spores belonging to Cladosporium and Alternaria genera were recorded. Of these spores, 5,103 were identified in 2000 and 5,128 in 2001. While 63.09% of the total spores were those of Cladosporium, 36.91% were of Alternaria. Relationships between airborne fungal spore presence and meteorological conditions were statistically investigated. A Shapiro-Wilk test revealed that the airborne Cladosporium and Alternaria spores differed from a normal distribution. Thus, a Friedmann test was performed followed by a Pearson Correlation Analysis. The effects of rainfall, temperature and wind speed on Cladosporium and Alternaria numbers were non-significant according to the sites and months (p > 0.05), but the effects of relative humidity on Cladosporium and Alternaria numbers were significant (p < 0.01). Spore concentrations reached to their highest levels in May 2001.