Introducing Alternaria tenuissima SBUp1, as an endophytic fungus of Ferula assa-foetida from Iran, which is a rich source of rosmarinic acid.

Endophytic fungi are the endogenous micro-organisms to interacting with the plant cells, which do not exhibit any symptoms on the host plant and may produce some of the main secondary metabolites of the host plant cells. Ferula assa-foetida is a perennial and endemic medicinal plant of Iran, which is a rich source of sesquiterpene, coumarins, polysulfides and phenolic acids. In this study, 28 endophytic fungi isolates including Fusarium (60·7%), Aspergillus (7·1%), Alternaria (17·9%) and Plectosphaerella (7·1%) were isolated from F. assa-foetida root (57·1%), stem (32·1%) and leaf (10·8%) collected from Parvand protected area. Subsequently, their ability to produce phenolic acids was evaluated. The high amounts of total phenol (326·09 mg g-1 of dry weight, DW), total flavonoid (901·11 mg g-1 DW) and antioxidant activity (247·96 mg l-1 ) were found in the supernatant fluid of SBUp1 isolate. The high-performance liquid chromatography analysis of 14 phenolic acids showed that rosmarinic acid (RA) is the main phenolic acid in the supernatant fluid of SBUp1 by 64·11 mg g-1 DW confirmed by the liquid chromatography coupled with mass spectrometric analysis. According to morphological identification followed by phylogenetic study based on internal transcribed spacer (ITS) sequencing (ITS1-5.8S-ITS2) analysis, the SBUp1 isolate was identified as Alternaria tenuissima. Eventually, to our knowledge, it is the first document confirming A. tenuissima as an endophytic fungus of F. assa-foetida, which is a rich source of RA.

A New Approach for Determining the Minimum Concentration of Proanthocyanidin for Preservation of Collagen in H Dentin.

PURPOSE: Introducing a minimum concentration and clinically relevant application time for grape seed extract (GSE) proanthocyanidin as a dentin preserver. MATERIALS AND METHODS: Dentin beams were demineralized in 10% phosphoric acid for 24h. Then, the following groups were prepared: G1: no treatment, G2: 2% CHX + 1min, G3: 1% GSE + 1min, G4: 1% GSE +5min, G5: 2% GSE + 1min, G6: 2% GSE + 5min, G7: 5% GSE + 1min and G8: 5% GSE + 5min. After 30 days, MALDI-TOF mass spectrometry was used to confirm the availability of digested peptide fragments and monitor the pattern of collagen digestion. Gravimetric assessment and HPLC-UV were utilized for quantitative measurement of dentin destruction. Glycine quantities were considered as measures of collagen digestion. RESULTS: 7% to 25% loss of dry mass was measured in experimental groups. Regarding the liberated Glycine, GSE dose- and time-responses were observed, so that, 5% GSE showed considerable protecting effect on collagen compared to 1 and 2% GSE (P⟨0.001) and 5min GSE application could establish superior dentin preservation compared to 1min application (P⟨0.001). CONCLUSION: 5-minute treatment of dentin at 2% GSE and above is essential for protecting the demineralized dentin collagen against biodegradation.

In vitro study of antioxidant and antibacterial activities of Lactobacillus probiotic spp.

This study investigated the influence of aeration and minimal medium conditions on antioxidant and antibacterial activities of 21 probiotic Lactobacillus strains isolated from dairy products. The probiotic potential of the isolates was evaluated by pH and bile tolerance. Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) was used to confirm the phenotypic identification of isolates. Antioxidant producer isolates were screened by resistance to reactive oxygen species (ROS). The antioxidant and antibacterial activities of extracellular materials after 48 h fermentation with antioxidative strains were determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and broth microdilution assays, respectively. The results indicate that the antioxidant capacity of supernatants was increased by using of both minimal medium and agitation. The antibacterial activity was increased in minimal medium, but there has nearly no change in the antibacterial properties by using both agitation and minimal medium. The maximum antibacterial activity was observed during mid-exponential phase until the beginning of the early-stationary phase, but the maximum antioxidant activity was detected at the stationary growth phase. There is a significant relationship between antioxidant and antibacterial activities of the cell-free probiotic extracts, and their production rates are closely related to the fermentation type. The bioactive materials from probiotics could be extracted in a large amount at an appropriate time under a suitable condition.

Identification of bacteria using volatile organic compounds.

The rapid diagnosis of respiratory infections has always been an important goal for medical professionals, because rapid and accurate diagnosis leads to proper and timely treatment, and consequently, reduces the costs of incorrect and long-term treatments, and antibiotic resistance. The present study was conducted with the aim of detecting volatile organic compounds (VOCs) in three bacteria: Pseudomonas aeruginosa, Acinetobacter baumannii, Klebsiella pneumoniae. Headspace of the studied bacteria, after separately culturing in two types of liquid medium in three different time-periods, was extracted by solid phase microextraction and analysed by gas chromatography mass spectrometry The analysis results of the VOCs produced by the studied bacteria indicate that some VOCs are common and some are unique in each bacterium. 1-penten-3-ol, levomenthol, and 2-octyl-1-ol for P. aeruginosa, cyclohexene, 4-ethenyl, and cis-Dihydro-α-terpinyl acetate for A. baumannii and 1,3-butadiene, butyraldehyde, longifolene, octyl acetate, tridecanol, dodecenal, (E)-2-hexyl ester, butanoic acid, and 5,5-dodecadinyl-1 12-diol for K. pneumoniae were identified as unique VOCs for each bacterium. Finally, it can be said that an accurate and rapid bacterial detection method can be achieved by using a tool that can detect bacterial VOCs. However, more studies are needed to design a tool for which all aspects have been assessed, so that it can give us a more complete pattern for the use of these compounds as biomarkers.

Polarity-based fractionation in proteomics: hydrophilic interaction vs reversed-phase liquid chromatography.

During recent decades, hydrophilic interaction liquid chromatography (HILIC) ahs been introduced to fractionate or purify especially polar solutes such as peptides and proteins while reversed-phase liquid chromatography (RPLC) is also a common strategy. RPLC is also a common dimension in multidimensional chromatography. In this study, the potential of HILIC vs RPLC chromatography was compared for proteome mapping of human peripheral blood mononuclear cell extract. In HILIC a silica-based stationary phase and for RPLC a C18 column were applied. Then separated proteins were eluted to an ion trap mass spectrometry system. Our results showed that the HILIC leads to more proteins being identified in comparison to RPLC. Among the total 181 identified proteins, 56 and 38 proteins were fractionated specifically by HILIC and RPLC, respectively. In order to demonstrate this, the physicochemical properties of identified proteins such as polarity and hydrophobicity were considered. This analysis indicated that polarity may play a major role in the HILIC separation of proteins vs RPLC. Using gene ontology enrichment analysis, it was also observed that differences in physicochemical properties conform to the cellular compartment and biological features. Finally, this study highlighted the potential of HILIC and the great orthogonality of RPLC in gel-free proteomic studies. Copyright © 2015 John Wiley & Sons, Ltd.