Volume 43, Issue 3

Background Study on the synthesis of kojic acid derivatives (KADs) in solvent-free system using scalable reactors and their biological activities is still lacking. Methods In this study, two types of KADs, were synthesized using saturated-fatty acid [lauric acid (LA)] and unsaturated-fatty acid [oleic acid (OA)] in stirred tank reactor (STR) and fluidized tank reactor (FTR). The yield and biological activities of the synthesized KADs were evaluated and compared. Results The highest yield of KADs (42.95%) was obtained in the synthesis using OA, with molar ratio of 1:1, enzyme loading of 5% (w/v), temperature of 70°C, using immobilized lipase N435 in STR. However, FTR may provide biocatalyst protection and reusability with reduced loss of KADs yield up to three cycles. In antioxidant assay, the hydroxyl-unsaturated-fatty acid of kojic acid (HUFA-KA) showed better activity as compared to hydroxyl-saturated-fatty acid of kojic acid (HSFA-KA) at concentrations ranging from 125 to 2000 μg/mL. In contrast, HSFA-KA showed better cytotoxicity effect against G361 melanoma cell as compared to HUFA-KA. Conclusion The yield of KADs obtained in STR was higher than that obtained in FTR. HUFA-KA could be used as potential lipophilic antioxidant while HSFA-KA has the potential to be used to treat melanoma skin disorder.

Objective The advent of resistant pathogenic microorganisms against current antimicrobial drugs prompted scientists to investigate novel molecules with new mechanisms. In this paper, some new 2-[2-[4-(ethyl/phenyl)cyclohexylidene]hydrazinyl]-4-(4-substitutedphenyl)thiazole ( 2a–2o ) derivatives were synthesized and studied for their antimicrobial activities. Materials and methods The title compounds ( 2a–2o ) were obtained via the reaction of 4-(ethyl/phenyl)cyclohexane-1-one with appropriate phenacyl bromide in ethanol at room temperature. The chemical structures of the compounds were elucidated by FT-IR, 1 H-NMR, 13 C-NMR, HRMS and elemental analysis. Antimicrobial activity of the compounds was measured by using broth microdilution method. Chloramphenicol and ketoconazole were used as reference drugs. Results Among the synthesized compounds, 2-[2-(4-phenylcyclohexylidene)hydrazinyl]-4-phenylthiazole ( 2h ) and 2-[2-(4-phenylcyclohexylidene)hydrazinyl]-4-(4-chlorophenyl)thiazole ( 2l ) have been found to exhibit potency almost four-fold better than ketoconazole against C. albicans with MIC 90 value of 1.95. Conclusion The current study contributed to the knowledge of the antimicrobial activity of thiazole bearing compounds.

Objective In the scope of presented work, synthesis of water based acrylate hydrogels, characterization, and their usage in controlled drug release systems were aimed to investigate. Methods Synthesis of acrylate based hydrogels that have different properties was carried out by free radical photopolymerization using photoinitiator. Because of its high biocompatibility, 2-hydroxyethyl metacrylate (HEMA) was used as monomer. Then drug release experiments were performed in pH 7.4 and 1.2 buffer solutions with certain ionic strength while the dynamic swelling behaviors were also determined. In the last part of the work, drug activities of synthesized drug-loaded hydrogels were tested in mediums containing Staphylococcus aureus and Pseudomonas aeruginosa bacteria cultures. Results ATR-FTIR spectrums of all synthesized hydrogels were analyzed. The characteristic O-H, C-H, C=O, C-O tension vibrations bands were observed in the spectrums of the hydrogels. The rate of drug release in acidic pH 1.2 for two types of hydrogels was observed to be much faster than at pH 7.4. It was determined that hydrogel swelling ratio decrease with increasing monomer ratio. All drug loaded hydrogels were effective to inhibit the growth of both two bacterial strains. Conclusion Hydrogels synthesized were found to be suitable for the controlled drug delivery applications.

Objective The aim of this work was to study the optimal cultivation conditions for β-galactosidase production by Bacillus licheniformis ATCC 12759. Materials and methods The screening of β-galactosidase production from B. licheniformis ATCC 12759 was performed by solid state fermentation method on media rich with rice bran (RB). Different factors were tested for the optimization of β-galactosidase production. Results Certain fermentation parameters involving incubation time, incubation temperature, inoculum level, moisture content, initial pH, agitation speed, size of fermentation medium and optimum temperature of β-galactosidase activity were studied separately. Maximal amount of β-galactosidase production was obtained when solid-state fermentation (SSF) was carried out using RB, having inoculum level 35%, moisture content of 20%, initial pH 7.5 at 37°C for 48 h. Conclusion Results indicated that optimal fermentation conditions play a key role in the maximum production of β-galactosidase from B. licheniformis ATCC 12759. This study shows the potential of the studied enzymes to be promoting candidates for the degradation of lactose and production of important bioproducts.

Objective Insects dwelled in soil play a key role in monitoring of metal contaminations. In order to explore the toxicity of lead (Pb) in soil, juvenile of oriental beetle Anomala corpulenta , were firstly applied to analyze effect of Pb. Methods In the current study, toxicity of different concentrations Pb on A. corpulenta in the laboratory was performed by measuring survival, growth and avoidance of animals. Meanwhile, activities of acetylcholinesterase (ChE), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in A. corpulenta were examined to quantify the extent of alterations caused by Pb treatments. Result Events of mortality were occurred in the Pb treated groups (1200 and 1600 mg Pb/kg soil). Administration of Pb treatments could result in a significant decrease of mean end weights in contrasted with that of control group. Treatment of Pb at 800 mg Pb/kg soil caused an obviously avoidance behavior. Expressions of ChE, SOD, CAT and GSH-Px of Pb treated groups were significant decreased compared with that of control group in the experiment observed. Conclusion Pb contamination of soil may cause multiple effects on juvenile A. corpulenta including occurrence of mortality, delay of growth and avoidance of contaminated environment.

Background Aim of the study was to produce α-amylase cheaply from Rhizoctonia solani AG-4 strain ZB-34 by solid-state fermentation and investigate its suitability for some industries. Methods Rhizoctonia solani AG-4 strain ZB-34 α-amylase was purified with starch affinity method. Results The best production of enzyme was achieved by using corn bran. Optimum pH and temperature were 5.5 and 50°C, respectively. V max and K 0.5 were determined as 238.8 U/mg protein and 0.03% from substrate-activity curve. Activity was maintained about 100% in the tested pHs after 1 day incubation. The enzyme conserved full of its activity at 4 and 28°C for 120 min. Mn 2+ , Ca 2+ , Tween 20, Triton X-100 and Triton X-114 activated the enzyme. The enzyme was highly active in the presence of some commercial detergents. The chocolate stains on the cotton fabrics were more effectively cleaned with the combination of a commercial detergent and purified enzyme. The purified enzyme also clarified the raw apple juice and desized the cotton fabrics. Conclusion The results showed that Rhizoctonia solani AG-4 strain ZB-34 α-amylase might have potential application as a detergent additive. In addition, its activity between pH 4.0 and 7.0 may facilitate its use in the food and fruit processing industries.

Background Selenium (Se 34 ) is an essential micronutrient for humans and animals and has growth promoting and antioxidative effects at low concentrations. Methods Effects of various sodium selenite (Na 2 SeO 3 ) doses on grain amino acid content of barley cultivars (Bülbül 89 and Çetin 2000) was investigated using ion exchange liquid chromatography. Results Majority of the amino acids could be altered with Selenium (Se) fertilization. Grain Se content of Bülbül 89 (0.175 mg kg −1 ) and Çetin 2000 (0.171 mg kg −1 ) were similar and both displayed an increase in proteinogenic, essential, and sulfur amino acids. The response of cultivars was more pronounced for Se accumulation and amino acid content at mid dose (12.5 mg ha −1 ). The quantities of proteinogenic, essential and sulfur amino acids increased considerably at that dose. Se induced increase in nitrogen content might cause an increase in some of the proteins of grain and consequently can alter amino acid composition. An obvious increase in the limiting amino acids (lysine and threonine) were prominent in response to Se fertilization. Conclusion Se treatment influence amino acid composition of barley grains; especially improve the quantity of limiting amino acids and consequently nutritional value of the grain.

Cotton ( Gossypium hirsutum L.) is the most important natural textile fiber crop grown worldwide. Several biotic and abiotic stress factors affect cotton yield due to lower genetic diversity for the traits of particular interest. Verticillium wilt (VW) is one of the major factors incurring huge cotton yield losses. The most effective management option against VW is the development of resistant cultivars. The resistant cultivars must also have superior fiber quality and yield traits. Therefore, the current study was aimed at screening some of the identified simple sequence repeats (SSR) markers for VW resistance and fiber quality traits of cotton genotypes in Turkey. Fifty different cultivars were screened with 30 SSR markers. Polymerase chain reaction (PCR) was conducted to amplify the SSR markers. The amplified bands were scored as 0 or 1 for absence and presence, respectively. For the molecular data analysis, polymorphism information content (PIC) values of molecular markers were calculated. Among tested SSR markers 13 were found polymorphic, which produced a total of 677 loci. The number of alleles per marker ranged from 1 to 4 and, overall average PIC values of markers ranged from 0.00 to 0.76, respectively. Principal component analysis executed on presence absence data yielded two distinct groups of cultivars screened. Hierarchical clustering revealed low genetic diversity among the tested cultivars. Based on the results TamcotCamdES, Gloria, Natalia, Lydia, Teks, Tamcot SP37H, N87 and BA525 are the promising cultivars for further breeding studies. The results of the current study also revealed that 4 SSR markers (DPL752 and DPL322 for fiber traits, DPL0022 and GH215 for VW resistance) out of 30 could be used for improving VW resistance and fiber quality in cotton through marker assisted selection.

Background Pectinases are one of the upcoming enzymes in food processing industries and they hydrolyze pectin substances. Objectives This study was done to examine the production of pectinase by Piriformospora indica in submerged fermentation (SmF) along with growth parameters. Materials and methods The fungus P. indica was cultured on Kafer medium supplemented with pectin for 0–12 days and fungus growth, number of spores, total protein content, and pectinase activity were investigated. Results Firstly, pectinase secretion by P. indica was confirmed by cup-plate assay. The maximum dry cell weight (10.21 g/L), growth yield (0.65 g/g), specific growth rate (0.56 day −1 ) and pectinase activity (10.47 U/mL) on pectin containing medium (P + ) were achieved after 6 day of culture. In the case of pectin free medium (P − ) all the parameters were less than P + medium. The pectinase production by P. indica on P + was 2.7 times higher than P − . The optimum pH and temperature for maximum polygalacturonase activity were 5 and 50°C, respectively. Conclusions For the first time, the work confirmed pectinase secretion by P. indica fungus and it could be a good source for pectinase production. Moreover, optimum pH 5, make it a potential candidate for future application in fruit juice industries.

Background Membrane bioreactor (MBR) systems used for wastewater treatment (WWT) processes are regarded as clean technologies. Degradation capacity of the predator bacterium, Bdellovibrio bacteriovorus , was used as a cleaning strategy for reducing membrane fouling. Method Wastewater with different sludge age and hydraulic retention time were filtered through Poly(ether)sulphone (PES) membranes using dead end reactor. Change in filtration performance after cleaning of membrane surface by B. bacteriovorus was measured by comparison of flux values. Bacterial community of the sludge was determined by 16SrRNA sequence analysis. Community profile of membrane surface was analyzed by fluorescent in situ hybridization technique. Results After cleaning of MP005 and UP150 membranes with predator bacteria, 4.8 L/m 2 ·h and 2.04 L/m 2 ·h increase in stable flux at steady state condition was obtained as compared to the control, respectively. Aeromonas, Proteus , and Alcaligenes species were found to be dominant members of the sludge. Bdellovibrio bacteriovorus lysed pure cultures of the isolated sludge bacteria successfully. FISH analysis of the membrane surface showed that Alfa-proteobacteria are the most numerous bacteria among the biofilm community on the membrane surface. Conclusion Results suggested that cleaning of MBR membranes with B. bacteriovorus has a potential to be used as a biological cleaning method.

Objective The percentage of large unstained cells (%LUCs) reflects peroxidase-negative cells and activated lymphocytes. Unlike other infections, the value of %LUCs in the diagnosis of fungal infections is not clear. We aimed to evaluate %LUCs and its correlations with other inflammatory parameters of invasive aspergillosis (IA) patients. Methods Twenty patients and 20 healthy participants were included. Full blood count parameters including %LUCs values were recorded. Platelet to lymphocyte ratio (PLR), neutrophil to lymphocyte ratio (NLR), monocyte to lymphocyte ratio (MLR), eosinophil to lymphocyte ratio (ELR) were calculated. Results There was a significant difference between the study groups for %LUCs [2.40 (2.22–3.25); 1.43 (1.25–2.10), respectively; p<0.001]. Furthermore, %LUCs were statistically significantly correlated with PLR, NLR and MLR (p=0.020, 0.040, 0.040; respectively) but not correlated with ELR (p>0.05). Conclusion The %LUCs values were significantly increased and correlated with markers of inflammation in patients. We suggest that the %LUCs is a useful predictor and may be an aid in the diagnosis and/or the management of IA and may help clinicians for follow up these patients in therapy process. Our study provides target pathways for further studies in the diagnosis of Aspergillus -infected patients using inflammatory parameters.

Aim This study aimed to isolate moderately halophilic bacteria from salted goat skins, to characterize these microorganisms and to determine their industrially important enzymes such as amylase, catalase, oxidase, caseinase, cellulase, DNase, lipase, lecithinase, protease, pullulanase, urease, phospholipase, xylanase and β-galactosidase. Methods Enzymes of these bacteria, isolated from skin samples belonging to eight countries and identified using phenotypic and genotypic methods, were examined in agar media. Results Thirty-nine isolates were fairly similar to species of genera Staphylococcus , Bacillus , Salinicoccus , Gracilibacillus , Chromohalobacter and Halomonas . Various carbon sources were utilized, and all isolates produced enzyme. Enzyme-producing species were Staphylococcus saprophyticus subsp. saprophyticus , Staphylococcus arlettae , Bacillus pumilus , Gracilibacillus dipsosauri , Salinicoccus roseus , Bacillus licheniformis , Chromohalobacter beijerinckii , Staphylococcus xylosus , Halomonas eurihalina , Staphylococcus equorum subsp. equorum , Halomonas zhanjiangensis , Halomonas venusta and Chromohalobacter canadensis . Fairly high percentage of isolates produced protease (87%) and catalase (100%). While more than 50% of isolates produced lipase (64%), β-galactosidase (59%) and oxidase (56%), less than 50% of isolates produced urease (46%), caseinase (28%), amylase (26%), lecithinase (8%) and cellulase (5%). Conclusion We detected that moderately halophilic bacteria on skins produced important enzymes, which may be used in diverse industrial applications in leather, feed, detergent, paper, food, chemical, medical, pharmaceutical, textile industries.

Background Lactic acid bacteria (LAB) are considered as GRAS (generally recognized as safe) and being used extensively as bio-preservatives. Bacteriocins, the metabolites of LAB, belong to a diverse family of naturally synthesized antimicrobial peptides. Objective Strain improvement for enhanced bacteriocin production by physical and chemical mutagenesis. Methods The bacterial strain was identified by 16S rDNA sequence analysis and improved by ultraviolet and ethidium bromide mutation. The resultant bacteriocin was biochemically characterized, purified and analyzed for mass determination. Results Among mutants of identified Lactobacillus acidophilus MS1, the UV 6 (ultraviolet mutant) revealed 3400 AU bacteriocin activity with 42% survival rate and EB 5 (ethidium bromide mutant) exhibited 4020 AU with 28% survival rate. Bacteriocin of 6.5 kDa was purified by cation exchange and gel exclusion chromatography. It was found to be thermally stable at 100°C for 30 min and maintained the stability up to 121°C. The activity was monitored in a wide range of pH (4–9). Conclusion Being resistant to several biochemical parameters, the bacteriocins have an effective incorporation in food, forage and pharmacy. There is a need to engage more efforts to explore novel bacteriocins and multifarious applications.

Objective The aim of this study was to evaluate the effects of hydraulic calcium silicate endodontic cements, MTA Angelus, MTA Fillapex, and Theracal LC, on erythrocyte and liver oxidative stress parameters of rats. Methods Right upper incisor of each rat was extracted and polyethylene tubes containing the dental cements, or left empty for the control group, were inserted into the extraction socket. Blood and liver samples of each animal were obtained after 7, 30, or 60 days. Thiobarbituric acid reactive substances (TBARS) levels and catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) activities were determined by spectrophotometry. Results Erythrocyte and liver TBARS levels, and CAT and SOD enzymatic activities were significantly increased in dental cement applied groups compared with controls on day 7. The highest erythrocyte and liver TBARS concentrations were observed in the MTA Angelus group on day 7 of exposure. On day 30, erythrocyte CAT activity remained markedly high, but the other parameters returned to almost normal levels. On day 60, all parameters were similar between the control and the experimental groups. Conclusions This is the first study to show that TBARS levels and antioxidant enzyme activities are transiently increased as a result of dental cement application.