Screening the antifungal activities of monoterpenes and their isomers against Candida species.

AIM: Increasing the occurrence of non-albicans Candida species with intrinsic or acquired resistance to antifungals as well as the emergence of multidrug Candida species coupled with the limited antifungal agents challenges the treatment of candidiasis. Consequently, a class of secondary metabolites of plants exhibiting decent antifungal activity. Therefore, this study aimed to evaluate the antifungal potential of various monoterpenes including Carvone, Limonene, Pinene, Menthone, Menthol, Camphor, Thujone, Citronellol, and Piperitone against standard and clinical isolates of Candida. METHODS AND RESULTS: Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of compounds were determined, using the broth Microdilution method based on M27-A3 protocol documented by clinical laboratory standard institute (CLSI). Amongst the tested monoterpenes, oxygenated terpenoids showed strong antifungal activity. Specifically, alcoholic terpenoids such as (±)-Citronellol possess more efficacy than the corresponding ketonic ones with MICs ranging from 0·03 to 2·00 µl ml-1 (0·16-10·80 mmol l-1 ). Among the examined yeasts, Candida tropicalis was the most susceptible species to (±)-Citronellol. Moreover, the examined monoterpenes successfully inhibited the growth of fluconazole-resistant Candida species. Moreover, statistical analysis showed no statistically significant difference between the (+) and (-) isomers, except for (±)-α-Pienene and (±) Menthone (ρ-value < 0·05). CONCLUSION: Among the tested monoterpenes, (±)-Citronellol was the most potent compounds followed by (+)-α-Pinene and Menthol. Considering the significant antifungal activity of the examined monoterpenes, they could be used in controlling or treating candidiasis. Those potent antifungal monoterpenes with GRAS status in addition to their pleasant taste and odour make them appropriate additive or preservative compounds in food and cosmetics products. Furthermore, these data might help researchers to predict EOs antifungal activities, after determining its constituents. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides new information about the antifungal activities of monoterpenes and their isomers presented widely in essential oils. Screening results against pathogenic yeasts confirm the correlation between the chemical structure of tested monoterpenes and their antifungal effects. The present findings might be helpful to anticipate the antifungal activity of essential oils.

Antibiotic resistance and frequency of class 1 integrons among Pseudomonas aeruginosa isolates obtained from wastewaters of a burn center in Northern Iran.

BACKGROUND: Pseudomonas aeruginosa, being responsible of a broad variety of infections, is considered an important nosocomial pathogen. The emergence of multiple-drug resistance among strains of P. aeruginosa appeared as a further public health concern. Due to the considerable ability of multiple-drug resistant P. aeruginosa strains to transmit themselves in the environment, we aimed to investigate the association of class 1 integrons with the antibiotic resistance profile of P. aeruginosa strains isolated from hospital wastewaters. METHODS: In this cross-sectional study, a total of 100 P. aeruginosa isolates were obtained from raw wastewater samples from February 2010 to January 2011 in a Teaching Burn Hospital in Guilan province. All isolates were identified as P. aeruginosa using standard microbiological tests. Antibiotic susceptibility was investigated using the disk diffusion method according to Clinical and Laboratory Standards Institute recommendations. All isolates were assayed for the presence of the class 1 integrons gene by PCR. RESULTS: Overall, 30 (30%) P. aeruginosa isolates were positive for the presence of class 1 integrons. The highest antibiotic resistance rates in both integron-positive and -negative isolates belonged to cephalexin and cephazolin, with 100% resistance. Amikacin and ciprofloxacin with the lowest level of resistance (13.3%) were the effective antibiotics against integron-positive isolates. The rates of MDR isolates were significantly higher among integron-positive isolates with 43.3% compared to negative isolates with 22.9% (P < 0.05). CONCLUSION: The results highlight the importance of class 1 integrons in multiple antibiotic resistance among P. aeruginosa isolates. Moreover, the spread of hospital derived wastewaters in the environment can be regarded as the origin of significant reservoirs for antibiotic-resistant pathogens.

Structure, Chaperone Activity, and Aggregation of Wild-Type and R12C Mutant αB-Crystallins in the Presence of Thermal Stress and Calcium Ion - Implications for Role of Calcium in Cataract Pathogenesis.

The current study was performed with the aim to evaluate the chaperoning ability, structural features, and aggregation propensity of wild-type and R12C mutant αB-crystallins (αB-Cry) under thermal stress and in the presence of calcium ion. The results of different spectroscopic analyses suggest that wild-type and mutant αB-Cry have dissimilar secondary and tertiary structures. Moreover, αB-Cry indicates slightly improved chaperone activity upon the R12C mutation. Thermal stress and calcium, respectively, enhance and reduce the extent of solvent-exposed hydrophobic surfaces accompanying formation of ordered and non-ordered aggregate entities in both proteins. Compared to the wild-type protein, the R12C mutant counterpart shows significant resistance against thermal and calcium-induced aggregation. In addition, in the presence of calcium, significant structural variation was accompanied by reduction in the solvent-exposed hydrophobic patches and attenuation of chaperone activity in both proteins. Additionally, gel mobility shift assay indicates the intrinsic propensity of R12C mutant αB-Cry for disulfide bridge-mediated protein dimerization. Overall, the results of this study are of high significance for understanding the molecular details of different factors that are involved in the pathomechanism of cataract disorders.

Positively charged imidazolium-based ionic liquid-protected silver nanoparticles: a promising disinfectant in root canal treatment.

AIM: To synthesize and characterize silver nanoparticles (Ag NPs) with different surface charges in order to evaluate their cytotoxicity and antibacterial activity in the absence and presence of dentine compared with NaOCl and CHX. METHODOLOGY: Ag NPs with positive, negative and neutral surface charges were synthesized and characterized. The first phase of the experiment determined the minimum inhibitory concentrations (MICs) of NPs against planktonic E. faecalis and compared them with that of NaOCl and CHX. The second phase tested the elimination of E. faecalis at different contact times (5, 20 and 60 min and 4 and 24 h), and the role of dentine in their inactivation was assessed. In the third phase, the most effective Ag NP solution was selected for cytocompatibility assessment. An MTT-based cytotoxicity assay was used to evaluate the cytotoxicity of the selected NP solution in different concentrations on L929 fibroblasts compared to that of 2.5% NaOCl and 0.2% CHX. Student's t-test and repeated measures manova approach were used for statistical analyses. RESULTS: The characterization revealed synthesis of colloidal NPs in the size range of 5-10 nm in diameter. The results indicated that Ag NP with a positive surface charge had the smallest MIC against planktonic E. faecalis, and it was active in very lower concentrations compared to NaOCl, CHX and the other tested AgNPs. Positive-charged Ag NPs at 5.7 × 10(-10)  mol L(-1) completely prevented the growth of E. faecalis after 5 min of contact time, a finding comparable to 0.025% NaOCl. Dentine powder had variable inhibitory effects on all tested materials after 1 h incubation period, but after 24 h, NaOCl and the positive-charged Ag NPs were not inhibited by dentine at any concentration used. CHX was the most and the positively charged Ag NP solution was the least toxic solutions to L929 fibroblasts (P < 0.001). CONCLUSIONS: Ag NP surface charge was important in bactericidal efficacy against E. faecalis. The positively charged imidazolium-based ionic liquid-protected Ag NPs showed promising antibacterial results against E. faecalis and exhibited a high level of cytocompatibility to L929 cells.

Microalgae biofuel potentials (review).

With the decrease of fossil based fuels and the environmental impact of them over the planet, it seems necessary to seek the sustainable sources of clean energy. Biofuels, is becoming a worldwide leader in the development of renewable energy resources. It is worthwhile to say that algal biofuel production is thought to help stabilize the concentration of carbon dioxide in the atmosphere and decrease global warming impacts. Also, among algal fuels' attractive characteristics, algal biodiesel is non toxic, with no sulfur, highly biodegradable and relatively harmless to the environment if spilled. Algae are capable of producing in excess of 30 times more oil per acre than corn and soybean crops. Currently, algal biofuel production has not been commercialized due to high costs associated with production, harvesting and oil extraction but the technology is progressing. Extensive research was conducted to determine the utilization of microalgae as an energy source and make algae oil production commercially viable.

Isolation and characterization of some moderately halophilic bacteria with lipase activity.

Lipases are an important class of enzymes which catalyze the hydrolysis of long chain triglycerides and constitute the most prominent group ofbiocatalysts for biotechnological applications. There are a number of lipases, produced by some halophilic microorganisms. In this study, some lipase producing bacteria from Maharlu salt lake located in south of Iran were isolated. All isolates were screened for true lipase activity on plates containing olive oil. The lipase activity was measured using titrimetric methods. Among thirty three isolates, thirteen strains demonstrating orange zone around colonies under UV light, were selected for identification using the molecular methods and some morphological characteristics. The bacterium Bacillus vallismortis BCCS 007 with 3.41 +/- 0.14 U/mL lipase activity was selected as the highest lipase producing isolate. This is the first report of isolation and molecular identification of lipase producing bacteria from Maharlu lake.

Characterization of hydrocortisone bioconversion and 16S RNA gene in Synechococcus nidulans cultures.

A unicellular cyanobacterium, Synechococcus nidulans (Pringsheim) Komárek, was isolated from paddy-fields and applied in the biotransformation experiment of hydrocortisone (1). This strain has not been previously tested for steroid bioconversion. Fermentation was carried out in BG-11 medium supplemented with 0.05% substrate at 25 degrees C for 14 days of incubation. The obtained products were chromatographically purified followed by their characterization using spectroscopic methods. 11beta,17beta-dihydroxyandrost-4-en-3-one (2), 11beta-hydroxyandrost-4-en-3,17-dione (3), and androst-4-ene-3,17-dione (4) were the main bioproducts in the hydrocortisone bioconversion. The observed bioreaction characteristics were the side chain degradation of the substrate to prepare compounds (2) and (3) following the 11beta-dehydroxylation for accumulation of the compound (4). Time course study showed the accumulation of the product (2) from the second day of the fermentation and compounds (3) and (4) from the third day. All the metabolites reached their maximum concentration in seven days. Cyanobacterial 16S rRNA gene was also amplified by PCR. Sequences were amplified using the universal prokaryotic primers which amplify a approximately 400-bp region of the 16S rRNA gene. PCR products were sequenced to confirm their authenticity as 16S rRNA gene of cyanobacteria. The result of PCR blasted with other sequenced cyanobacteria in NCBI showed 99% identity to the 16S small subunit rRNA of seven Synechococcus species.

Analysis of antimicrobial, antifungal and antioxidant activities of Juniperus excelsa M. B subsp. Polycarpos (K. Koch) Takhtajan essential oil.

Juniperus excelsa M.B subsp. Polycarpos (K.Koch), collected from south of Iran, was subjected to hydrodistillation using clevenger apparatus to obtain essential oil. The essential was analyzed by gas chromatography/mass spectrometry (GC/MS) and studied for antimicrobial, antifungal and antioxidant activities. The results indicated α-pinene (67.71%) as the major compound and α-cedral (11.5%), δ3-carene (5.19%) and limonene (4.41%) in moderate amounts. Antimicrobial tests were carried out using disk diffusion method, followed by the measurement of minimum inhibitory concentration (MIC). All the Gram positive and Gram negative bacteria were susceptible to essential oil. The oil showed radical scavenging and antioxidant effects.