The encapsulation of living cells in materials with good optical and mechanical properties often produces death or stress due to the release of toxic byproducts originated during the synthesis. We present here a method to assess the cellular stress that silica entrapment exerts over living cells taking into account the main preparation variables such as the nature of the silica source, protecting functional groups, total solid concentration, or indirect procedures. Measurement of the cellular stress status of genetically modified Sacharomyces cereVisiae, a true biological probe, allowed us to perform a quantitative analysis of cellular stress in a short time basis (compared to conventional long-term viability tests), opening the gate for a more sophisticated approach to optimize the synthesis conditions. In addition, the aforementioned findings allowed the preparation of novel materials with enhanced optical and mechanical properties. The relation of cellular stress with initial viability is also discussed.
d-aminolevulinic acid, the precursor of porphyrin biosynthesis has been used to induce the endogenous synthesis of the photosensitiser protoporphyrin IX for photodynamic therapy in the treatment of various tumours. The aim of this work was to characterise the d-aminolevulinic acid transport system in the murine mammary adenocarcinoma cell line LM3 using 14 C-daminolevulinic acid, to finally improve d-aminolevulinic acid incorporation in mammalian cells. Our results showed that daminolevulinic acid is incorporated into these cells by two different mechanisms, passive diffusion which is important at the beginning of the incubation, and active transport. Specificity assays suggested that the transporter involved in d-aminolevulinic acid incorporation is a BETA transporter, probably GAT-2.
The Saccharomyces cerevisiae UGA4 gene encodes a permease capable of importing ␥-aminobutyric acid (GABA) and ␦-aminolevulinic acid (ALA) into the cell. GABA-dependent induction of this permease requires at least two positive-acting proteins, the specific factor Uga3 and the pleiotropic factor Uga35/Dal81. UGA4 is subjected to a very complex regulation, and its induction is affected by the presence of extracellular amino acids; this effect is mediated by the plasma membrane amino acid sensor SPS. Our results show that leucine affects UGA4 induction and that the SPS sensor and the downstream effectors Stp1 and Stp2 participate in this regulation. Moreover, we found that the Uga3 and Uga35/Dal81 transcription factors bind to the UGA4 promoter in a GABA-dependent manner and that this binding is impaired by the presence of leucine. We also found that the Leu3 transcription factor negatively regulates UGA4 transcription, although this seems to be through an indirect mechanism.
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