Design and introduction of a rational mechanical eluting system for leukocyte recovery from leukoreduction filters: A cell differential approach.

OBJECTIVES: The use of leukoreduction filters has been highly increased in Iranian Blood Transfusion Centers within the last decade to provide sufficient leukoreduced blood products from healthy repeated donors for alloimmunized or sensitive recipients. Leucoflex LCR5, the dominant brand which procured by the Iranian Blood Transfusion Organization, is the most updated generation of the filters used around the world. MATERIAL AND METHODS: In this study, we recovered trapped leukocytes from these filters using different buffer solutions and optimized elution method. The count of recovered cells assessed by cell counter, and cell viability was detected using trypan blue staining. The percent of leukocyte subpopulations was evaluated using a panel of monoclonal antibodies and flow cytometric analysis. RESULTS: It illustrated that a buffer solution consistent with PBS in pH 7.2 containing 2mM EDTA and 4% (w/w) Dextran 40 was the best buffer for LCR5 filter backflushing. The white cell counted as 4.56×108 Granulocytes, 3.34×108 Lymphocytes, and 0.64×108 Monocytes according to analysis with auto hemoanalysis and flow cytometric methods. CONCLUSION: The study guides and assists blood management systems in arranging a national blood profile database for future cell therapy strategies. Also, the recovered cells could be of significance in stem cell research, cellular interaction studies as well as novel molecular developments in drug discovery.

Increasing the antibacterial activity of gentamicin in combination with extracted polyphosphate from Bacillus megaterium.

AIMS: The aim of this research was production of polyphosphate (poly P) and study on its antibacterial effects. METHODS AND RESULTS: Poly P granules in the cells were observed with the help of Albert staining and extracted by Mussig-Zufika method. Thin layer chromatography and nuclear magnetic resonance spectroscopy ((31) P NMR) were used to characterize properties of these granules. Relation of phosphorus consumption and poly P production with growth was determined by the vanado-molybdate colorimetric method. Among the 60 strains of bacteria isolated from the environmental samples, strain G11 showed ability for the formation of high levels of poly P. Phylogenetic analysis showed that this isolate had 98% similarity with Bacillus megaterium. 16S rRNA sequence of isolate was deposited in GenBank with accession number JX115009. The average poly P chain length was 10·5 in this bacterium. The antimicrobial activity of bacterial extracted poly P was much better than chemical poly P, and its interaction with gentamicin increased the activity of this drug. The best synergistic activity of this interaction was observed for Corynebacterium glutamicum and Pseudomonas aeruginosa species. The highest adsorption of phosphorus occurred in stationary phase of growth curve, and then the amount of phosphorus increased in medium by degradation of stored poly P. CONCLUSIONS: In this study, we isolated a high-level producer bacterium of poly P and extracted poly P by chemical treatment. In addition, we compared antimicrobial activity of chemical poly P with bacterial poly P and its interaction with gentamicin against both Gram-positive and Gram-negative bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: Many studies have shown that bacteria are becoming resistant to gentamicin sulphate. In this study, we approved that Acinetobacter baumannii, a pathogenic gentamicin-resistant bacterium, is sensitive to bacterial poly P, and thus, this poly P can be substituted for gentamicin in treatment.

A simple method for primary screening of antibacterial peptides in plant seeds.

BACKGROUND AND OBJECTIVES: Regarding the importance of finding new antibacterial drugs, screening of plants as a promising resource are now conducted worldwide. In this study, we report the application of a simple previously described method for screening of different plant seeds in order to find the best resources of plant antimicrobial peptides. MATERIALS AND METHODS: Total water soluble protein of 10 different plant seeds were extracted and subjected to SDS-PAGE and subsequent agar-overlay bioassays. Standard strains of Staphylococcus aureus, Enterococcus faecium and Escherichia coli were included in the bioassays. This method also was used for total proteins precipitated by Ammonium sulphate which ensure the protein nature of the test substances. Molecular size and the amounts of effective peptides were estimated using Tricin-SDS-PAGE and densitometry. RESULTS: Two different plant seeds showed noticeable antibacterial activities against tested Gram positive bacteria and a moderate inhibitory effect on Gram negative ones. Based on the results of Tricin-SDS-PAGE analysis which were carried out in parallel to bioassays, it was concluded that effective antibacterial substances are peptides with molecular weight of slightly larger than 5 kDa. CONCLUSION: On the basis of results of agar-overlay experiments and by screening of 10 different herbal seeds, we could introduce seeds of M. sativa L. and Onobrychis sativa Lam., as great sources of putative plant antibacterial peptides. The proposed screening method can be used for screening of large number of different plant seeds and even other parts of the plant body, regarding some necessary modification in total water soluble protein extraction steps.

The utilization of aniline, chlorinated aniline, and aniline blue as the only source of nitrogen by fungi in water.

The ability of fungi to degrade aniline and its derivatives in water is reported. Several fungi are able to degrade aniline and its derivatives as a sole nitrogen, carbon and energy source. Some of these fungi were obtained from activated sludge by enrichment technique. Among the 10 studied fungi, Fusarium sp. and Rhizopus sp. utilize aniline as a sole nitrogen, carbon and energy source, with production of acetanilide and catechol. Fusarium sp. utilized 70% of 10 mmol aniline and produced 3.55 mM ammonia during 30 days. Rhizopus sp. utilized 65% of 10 mmol aniline during 30 days. Rhizopus sp. and Fusarium sp. utilized only 2-chloroaniline and 3-chloroaniline as nitrogen source in the presence of glucose, with production of catechol, ammonium and chloride. The utilization of 2-chloroaniline was better than 3-chloroaniline, by Fusarium sp. and Rhizopus sp. Cladosporium sp. was the best isolate which could use aniline blue as the only source of nitrogen. This fungus reduced 89% of aniline blue, and ammonia is produced as the result of aniline blue biodegradation by Cladosporium sp.

Biodegradation of lignocellulosic waste by Aspergillus terreus.

Biodegradation of lignocellulosic waste by Aspergillus terreus is reported for the first time. This isolate produced 250 CMCase (carboxymethyl cellulase or endoglucanase) U.ml(-1) and biodegraded hay and straw during 3 days and the biomass production on straw was 5g.L(-1) dry weight from 0.25 cm2 inoculated mycellium. This strain secreted endocellulases and exocellulases in the culture medium, but some of the enzymes produced, remained cell membrane bound. Cell bound enzymes were released by various treatments. The highest amount of endoglucanase and exoglucanase was released when the cells were treated with sonication. Aspergillus terreus was added to two tanks containing sugar wastewater and pulp manufacturing waste, as a seed for COD removal. This fungus reduced the COD by 40-80 percent, also, ammonia was reduced from 14.5 mM to 5.6 mM in sugar beet wastewater. The effects of crude enzyme of this fungus for COD removal was studied.