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Unicellular green algae, like Chlorella, offer a potentially useful system for the expression of heterologous proteins. However, the development of Chlorella as a bioreactor has been delayed owing to the lack of a stable transformation technique. Here we report on the use of micro-projectile bombardment to introduce the nitrate reductase (NR) gene from Chlorella vulgaris into NR-deficient Chlorella sorokiniana mutants, resulting in stable transformants. The stable transformants were able to grow on nitrate medium after repeated passages between selective and nonselective medium and exhibited inducible nitrate reductase activity comparable to that of wild-type cells. Southern analysis suggests homologous recombination occurs with insertion of the wild type gene into the mutated gene and that the genes of the two Chlorellaspecies used are very similar. Specific RNase protection assays, selecting for a poorly conserved region of the gene, identified the presence of the C. vulgaris NR transcript only in the transformed C. sorkiniana mutant and not in the mutant. [ABSTRACT FROM AUTHOR]

At present, there are no proven pharmacological treatments demonstrated to improve long term functional outcomes following traumatic brain injury(TBI). In the setting of non-penetrating TBI, sterile brain inflammatory responses are associated with the development of cerebral edema, intracranial hypertension, and secondary neuronal injury. There is increasing evidence that endogenous apolipoprotein E(apoE) modifies the neuroinflammatory response through its role in downregulating glial activation, however, the intact apoE holoprotein does not cross the blood-brain barrier due to its size. To address this limitation, we developed a small 5 amino acid apoE mimetic peptide(CN-105) that mimics the polar face of the apoE helical domain involved in receptor interactions. The goal of this study was to investigate the therapeutic potential of CN-105 in a murine model of closed head injury. Treatment with CN-105 was associated with a durable improvement in functional outcomes as assessed by Rotarod and Morris Water Maze and a reduction in positive Fluoro-Jade B stained injured neurons and microglial activation. Administration of CN-105 was also associated with reduction in mRNA expression of a subset of inflammatory and immune-related genes. [ABSTRACT FROM AUTHOR]