A genomic collection of haploid Saccharomyces cerevisiae deletion strains provides a unique resource for systematic analysis of gene interactions. Double-mutant haploid strains can be constructed by the synthetic genetic array (SGA) method, wherein a query mutation is introduced by mating to mutant arrays, selection of diploid double mutants, induction of meiosis, and selection of recombinant haploid doublemutant progeny. The mechanism of haploid selection is mating-type-regulated auxotrophy (MRA), by which prototrophy is restricted to a particular haploid genotype generated only as a result of meiosis. MRA escape leads to false-negative genetic interaction results because postmeiotic haploids that are supposed to be under negative selection instead proliferate and mate, forming diploids that are heterozygous at interacting loci, masking phenotypes that would be observed in a pure haploid double-mutant culture. This work identified factors that reduce MRA escape, including insertion of terminator and repressor sequences upstream of the MRA cassette, deletion of silent mating-type loci, and utilization of a-type instead of a-type MRA. Modifications engineered to reduce haploid MRA escape reduced false negative results in SGA-type analysis, resulting in .95% sensitivity for detecting gene-gene interactions.

In the last decade, nerve tissue engineering has attracted much attention due to the incapability of self-regeneration. Nerve tissue regeneration is mainly based on scaffold induced nanofibrous structures using both bio and synthetic polymers. The produced nanofibrous scaffolds have to be similar to the natural extracellular matrix and should provide an appropriate environment for cells to attach onto. Nanofibrous scaffolds can support or regenerate cells of tissue. Electrospinning is an ideal method for producing the nanofibrous scaffolds. In this study, Bacterial cellulose (BC)/ Poly (ε-caprolactone) (PCL) blend nanofibrous scaffolds were successfully prepared by electrospinning for nerve tissue induced repair. The produced nanofibrous scaffolds contain well defined interconnected nanofiber networks with hollow micro/nanobeads. Firstly, in-vitro biocompatibilities of nanofibrous scaffolds were tested with L2929 murine fibroblasts and improved cell adhesion and proliferation was observed with polymer blends compared with PCL only. The primary cell culture was performed with dorsal root ganglia (DRG) cells on nanofibrous samples and the samples were found suitable for enhancing neural growth and neurite outgrowth. Based on these results, the BC/PCL (50:50 wt. %) nanofibrous scaffolds exhibited nerve-like branching and are excellent candidate for potential biomimetic applications in nerve tissue engineering regeneration.

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ABSTRACT:The antimicrobial polymer/polymer macrocomplexes were synthesized by radical alternating copolymerization of N-vinyl-2-pyrrolidone with maleic anhydride [poly(VP-alt-MA)] with 2,2 0 -azobis-isobutyronitrile as an initiator at 658C in dioxane solutions under nitrogen atmosphere, and interaction of prepared copolymer with poly(ethylene imine) (PEI) in aqueous solutions. The susceptibility of some Gram-negative (Salmonella enteritidis and Escherichia coli) and Gram-positive (Staphylococcus aureus and Listeria monocytogenes) bacteria to the alternating copolymer and its PEI macrocomplexes with different compositions in microbiological medium was studied using pour-plate technique. All the studied polymers, containing biologically active moieties in the form of ionized cyclic amide, and macrobranched aliphatic amine groups and acid/amine complexed fragments, were more effective against L. monocytogenes than those for Gram-positive S. aureus bacterium. This fact was explained by different surface layer structural architectures of biomacromolecules of tested bacteria. The resulting polymeric antimicrobial materials are expected to be used in various areas of medicine and food industry.