Project description:Objective: 2-Hydroxyethyl methacrylate (HEMA), one of the most common components of tooth filling materials, could diffuse throughout dentine, and in gingival and pulp cells, affecting odontoblast vitality. Design: the aim of this study was to assess the differential transcriptome modulation induced by HEMA treatment in human cultured gingival fibroblasts (HGFs) at different exposure time compared to untreated cells. Materials and methods: Gene expression profile was performed by employing a whole gene microarray approach on RNA extracted from cultured HGFs exposed to 3 mmol/l HEMA for 24 or 96 h. Differentially identified transcripts were analyzed by Ingenuity Pathways Analysis software to disclose genes biological functions. Results: Hierarchical clustering analysis of the data set originated from a total of 8 experiments showed the selective presence of two gene clusters, composed by hundreds trandcripts differentially expressed after 24-h and 96-h HEMA treatment. when compared to untreated controls. Functional analysis showed the transcripts involvement mainly in cell survival, proliferation and death, with an unbalance towards cell growth and inflammatory response. Conclusions: Data analysis showed an overall damage induced by HEMA exposure for both HGFs cultures at 24 and 96-h mainly leading to a proliferation impairement. Interestingly 24-h HEMA exposure could induce the cells to trigger repair mechanisms evidencing an early compensatory response and survive while. 96-h incubation might increase apoptosis as a consequence of the chronic damage. Gene expression profile of Human Gingival Fibroblasts (HGFs) exposed to 3mM 2-Hydroxyethyl Methacrylate (HEMA) for 24- and 96-h versus untreated Human Gingival Fibroblasts at 24- and 96-h. A total of 8 experiments was carried out including biological and technical replicates.
Project description:Objective: 2-Hydroxyethyl methacrylate (HEMA), one of the most common components of tooth filling materials, could diffuse throughout dentine, and in gingival and pulp cells, affecting odontoblast vitality. Design: the aim of this study was to assess the differential transcriptome modulation induced by HEMA treatment in human cultured gingival fibroblasts (HGFs) at different exposure time compared to untreated cells. Materials and methods: Gene expression profile was performed by employing a whole gene microarray approach on RNA extracted from cultured HGFs exposed to 3 mmol/l HEMA for 24 or 96 h. Differentially identified transcripts were analyzed by Ingenuity Pathways Analysis software to disclose genes biological functions. Results: Hierarchical clustering analysis of the data set originated from a total of 8 experiments showed the selective presence of two gene clusters, composed by hundreds trandcripts differentially expressed after 24-h and 96-h HEMA treatment. when compared to untreated controls. Functional analysis showed the transcripts involvement mainly in cell survival, proliferation and death, with an unbalance towards cell growth and inflammatory response. Conclusions: Data analysis showed an overall damage induced by HEMA exposure for both HGFs cultures at 24 and 96-h mainly leading to a proliferation impairement. Interestingly 24-h HEMA exposure could induce the cells to trigger repair mechanisms evidencing an early compensatory response and survive while. 96-h incubation might increase apoptosis as a consequence of the chronic damage.
Project description:Gene expression profiling of immortalized human mesenchymal stem cells with hTERT/E6/E7 transfected MSCs. hTERT may change gene expression in MSCs. Goal was to determine the gene expressions of immortalized MSCs.
Project description:Transcriptional profiling of human mesenchymal stem cells comparing normoxic MSCs cells with hypoxic MSCs cells. Hypoxia may inhibit senescence of MSCs during expansion. Goal was to determine the effects of hypoxia on global MSCs gene expression.
Project description:Kynureninase is a member of a large family of catalytically diverse but structurally homologous pyridoxal 5'-phosphate (PLP) dependent enzymes known as the aspartate aminotransferase superfamily or alpha-family. The Homo sapiens and other eukaryotic constitutive kynureninases preferentially catalyze the hydrolytic cleavage of 3-hydroxy-l-kynurenine to produce 3-hydroxyanthranilate and l-alanine, while l-kynurenine is the substrate of many prokaryotic inducible kynureninases. The human enzyme was cloned with an N-terminal hexahistidine tag, expressed, and purified from a bacterial expression system using Ni metal ion affinity chromatography. Kinetic characterization of the recombinant enzyme reveals classic Michaelis-Menten behavior, with a Km of 28.3 +/- 1.9 microM and a specific activity of 1.75 micromol min-1 mg-1 for 3-hydroxy-dl-kynurenine. Crystals of recombinant kynureninase that diffracted to 2.0 A were obtained, and the atomic structure of the PLP-bound holoenzyme was determined by molecular replacement using the Pseudomonas fluorescens kynureninase structure (PDB entry 1qz9) as the phasing model. A structural superposition with the P. fluorescens kynureninase revealed that these two structures resemble the "open" and "closed" conformations of aspartate aminotransferase. The comparison illustrates the dynamic nature of these proteins' small domains and reveals a role for Arg-434 similar to its role in other AAT alpha-family members. Docking of 3-hydroxy-l-kynurenine into the human kynureninase active site suggests that Asn-333 and His-102 are involved in substrate binding and molecular discrimination between inducible and constitutive kynureninase substrates.
Project description:Single-nucleus RNA sequencing (snRNA-seq) was used to profile the transcriptome of 16,015 nuclei in human adult testis. This dataset includes five samples from two different individuals. This dataset is part of a larger evolutionary study of adult testis at the single-nucleus level (97,521 single-nuclei in total) across mammals including 10 representatives of the three main mammalian lineages: human, chimpanzee, bonobo, gorilla, gibbon, rhesus macaque, marmoset, mouse (placental mammals); grey short-tailed opossum (marsupials); and platypus (egg-laying monotremes). Corresponding data were generated for a bird (red junglefowl, the progenitor of domestic chicken), to be used as an evolutionary outgroup.