Optimized Isolation of Extracellular Vesicles From Various Organic Sources Using Aqueous Two-Phase System.

From biomarkers to drug carriers, Extracellular Vesicles (EVs) are being used successfully in numerous applications. However, while the subject has been steadily rising in popularity, current methods of isolating EVs are lagging behind, incapable of isolating EVs at a high enough quantity or quality while also requiring expensive, specialized equipment. The "isolation problem" is one of the major obstacles in the field of EV research - and even more so for their potential, widespread use for clinical diagnosis and therapeutic applications. Aqueous Two-Phase Systems (ATPS) has been reported previously as a promising method for isolating EVs quickly and efficiently, and with little contaminants - however, this method has not seen widespread use. In this study, an ATPS-based isolation protocol is used to isolate small EVs from plant, cell culture, and parasite culture sources. Isolated EVs were characterized in surface markers, size, and morphological manner. Additionally, the capacity of ATPS-based EV isolation in removing different contaminants was shown by measuring protein, fatty acid, acid, and phenol red levels of the final isolate. In conclusion, we have shown that EVs originating from different biological sources can be isolated successfully in a cost-effective and user-friendly manner with the use of aqueous two-phase systems.

Cytotoxicity, bactericidal and hemostatic evaluation of oxidized cellulose microparticles: Structure and oxidation degree approach.

Oxidized cellulose is the most used hemostatic materials in clinical applications. In addition to its perfect hemostatic efficiency, it is degradable under in vivo conditions and supremely prevents bacterial growth. On the other hand, one of the drawbacks of the oxidized cellulose is cytotoxicity due to the strongly acidic nature during degradation. There is a number of commercially available oxidized cellulose products which are derived from regenerated and non-regenerated cellulose. On the other hand, the effect of oxidation degree and structure (regenerated or non-regenerated) on product efficiency is undetermined. Moreover, oxidation degree which is primary factor for both bactericidal and hemostatic efficiency is also crucial for assessment of the product. In this study, oxidized cellulose versus oxidized regenerated cellulose microparticles with various oxidation degree was produced and characterized. Comparative studies were conducted in terms of bactericidal and hemostatic efficiencies in addition to cytotoxicity. The results could be a reference for the optimized oxidized cellulose product for the hemostatic applications.

Anti-microbial coating innovations to prevent infectious diseases (AMiCI): Cost action ca15114.

Worldwide, millions of patients are affected annually by healthcare-associated infection (HCAI), impacting up to 80,000 patients in European Hospitals on any given day. This represents not only public health risk, but also an economic burden. Complementing routine hand hygiene practices, cleaning and disinfection, antimicrobial coatings hold promise based, in essence, on the application of materials and chemicals with persistent bactericidal or -static properties onto surfaces or in textiles used in healthcare environments. The focus of considerable commercial investment and academic research energies, such antimicrobial coating-based approaches are widely believed to have potential in reduction of microbial numbers on surfaces in clinical settings. This belief exists despite definitive evidence as to their efficacy and is based somewhat on positive studies involving, for example, copper, silver or gold ions, titanium or organosilane, albeit under laboratory conditions. The literature describes successful delay and/or prevention of recontamination following conventional cleaning and disinfection by problematic microbes such as methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin resistant enterococci (VRE), among others. However, there is a scarcity of studies assessing antimicrobial surfaces other than copper in the clinical environment, and a complete lack of published data regarding the successful implementation of these materials on clinically significant outcomes (including HCAI). Through its Cooperation in Science and Technology program (COST), the European Commission has funded a 4-year initiative to establish a network of stakeholders involved in development, regulation and use of novel anti-microbial coatings for prevention of HCAI. The network (AMiCI) comprises participants of more than 60 universities, research institutes and companies across 29 European countries and, to-date, represents the most comprehensive consortium targeting use of these emergent technologies in healthcare settings. More specifically, the network will prioritise coordinated research on the effects (both positive and negative) of antimicrobial coatings in healthcare sectors; know-how regarding availability and mechanisms of action of (nano)-coatings; possible adverse effects of such materials (e.g., potential emergence of microbial resistance or emission of toxic agents into the environment); standardised performance assessments for antimicrobial coatings; identification and dissemination of best practices by hospitals, other clinical facilities, regulators and manufacturers.

Intra- and inter-species interactions within biofilms of important foodborne bacterial pathogens.

A community-based sessile life style is the normal mode of growth and survival for many bacterial species. Under such conditions, cell-to-cell interactions are inevitable and ultimately lead to the establishment of dense, complex and highly structured biofilm populations encapsulated in a self-produced extracellular matrix and capable of coordinated and collective behavior. Remarkably, in food processing environments, a variety of different bacteria may attach to surfaces, survive, grow, and form biofilms. Salmonella enterica, Listeria monocytogenes, Escherichia coli, and Staphylococcus aureus are important bacterial pathogens commonly implicated in outbreaks of foodborne diseases, while all are known to be able to create biofilms on both abiotic and biotic surfaces. Particularly challenging is the attempt to understand the complexity of inter-bacterial interactions that can be encountered in such unwanted consortia, such as competitive and cooperative ones, together with their impact on the final outcome of these communities (e.g., maturation, physiology, antimicrobial resistance, virulence, dispersal). In this review, up-to-date data on both the intra- and inter-species interactions encountered in biofilms of these pathogens are presented. A better understanding of these interactions, both at molecular and biophysical levels, could lead to novel intervention strategies for controlling pathogenic biofilm formation in food processing environments and thus improve food safety.

Effectiveness of organic acid, ozonated water and chlorine dippings on microbial reduction and storage quality of fresh-cut iceberg lettuce.

BACKGROUND: The comparative effects of organic (citric and lactic) acids, ozone and chlorine on the microbiological population and quality parameters of fresh-cut lettuce during storage were evaluated. RESULTS: Dipping of lettuce in 100 mg L(-1) chlorine solution reduced the numbers of mesophilic and psychrotrophic bacteria and Enterobacteriaceae by 1.7, 2.0 and 1.6 log(10) colony-forming units (CFU) g(-1) respectively. Treatment of lettuce with citric (5 g L(-1)) and lactic (5 mL L(-1)) acid solutions and ozonated water (4 mg L(-1)) reduced the populations of mesophilic and psychrotrophic bacteria by 1.7 and 1.5 log(10) CFU g(-1) respectively. Organic acid dippings resulted in lower mesophilic and psychrotrophic counts than ozonated water and chlorine dippings during 12 days of storage. Lactic acid dipping effectively reduced (by 2.2 log(10) CFU g(-1)) and maintained low populations of Enterobacteriaceae on lettuce for the first 6 days of storage. No significant (P > 0.05) changes were observed in the texture and moisture content of lettuce samples dipped in chlorine, organic acids and ozonated water during storage. Colour, ß-carotene and vitamin C values of fresh-cut iceberg lettuce did not change significantly (P > 0.05) until day 8. CONCLUSION: Lactic and citric acid and ozonated water dippings could be alternative treatments to chlorine dipping to prolong the shelf life of fresh-cut iceberg lettuce. Copyright © 2007 Society of Chemical Industry.

Modeling the growth kinetics of Bacillus cereus as a function of temperature, pH, sodium lactate and sodium chloride concentrations.

Mathematical models describing the growth kinetic parameters (lag phase duration and growth rate) of Bacillus cereus as a function of temperature, pH, sodium lactate and sodium chloride concentrations were obtained in this study. In order to get a residual distribution closer to a normal distribution, the natural logarithm of the growth kinetic parameters were used in modeling. For reasons of parsimony, the polynomial models were reduced to contain only the coefficients significant at a level of p