Project description:To study the effect of Plasmodium falciparum-infected erythrocytes on gene expression in NK92 cells, microarray analysis after 6, 12 and 24 hours of co-culture with either uRBC or iRBC was performed. The aim was to identify pathways in NK92 cells that are switched on after iRBC encounter in a time-dependent manner that will help to understand the mechanisms in innate immune defenses against Plasmodium falciparum infection.
Project description:To study the effect of Plasmodium falciparum-infected erythrocytes on gene expression in NK92 cells, microarray analysis after 6, 12 and 24 hours of co-culture with either uRBC or iRBC was performed. The aim was to identify pathways in NK92 cells that are switched on after iRBC encounter in a time-dependent manner that will help to understand the mechanisms in innate immune defenses against Plasmodium falciparum infection. Variation in gene expression of NK92 cells was determined after 6, 12, and 24 hours of co-culture with either infected or uninfected RBC compared to timepoint 0 (start of co-culture, untreated control). All experiments were done in triplicate, that means that samples were collected in 3 different experiments (A, B, and C) each time after 0, 6, 12 and 24 hours of co-culture.
Project description:Whole blood RNA-Seq was applied to investigate gene expression kinetics in Tanzanian males who underwent controlled malaria infection by intradermal injection with aseptic, purified, cryopreserved Plasmodium falciparum sporozoites.
Project description:The aim of the study was to determine the effect of natural killer (NK) cells on the global gene expression in Plasmodium falciparum. FCR3-CSA-infected red blood cells (RBCs) were co-cultured for 24h with NK92. Prior to RNA extraction, the parasitized RBCs were separated via Ficoll from the lymphocyte fraction. Control cultures of parasites without NK cells were cultivated in parallel. In addition, the effect of the separation method was determined via a Ficoll control culture of FCR3-CSA that was passed over a Ficoll gradient. All three treatments were performed in triplicate. The RNA was reverse-transcribed and hybridized onto microarrays.
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:Healthy Kenyan children with (n=10) or without (n=14) a previous history of complicated Plasmodium falciparum infection had aliquots of their whole-blood cultured ex-vivo and either mock infected or infected with Plasmodium falciparum (A4 strain) for 5 (n=24) and 9 hours (n=11). A clinical history design type is where the organisms clinical history of diagnosis, treatments, e.g. vaccinations, surgery etc. Time: lenght of co-culture Infection: whole-blood cells cultured ex-vivo in the presence of either uninfected red blood cells or red blood cells infected with Plasmodium falciparum Disease State: Healthy individuals with (PrevHxMal) or without (NoHxMal) previous exposure to complicated Plasmodium falciparum infection
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:Healthy Kenyan children with (n=10) or without (n=14) a previous history of complicated Plasmodium falciparum infection had aliquots of their whole-blood cultured ex-vivo and either mock infected or infected with Plasmodium falciparum (A4 strain) for 5 (n=24) and 9 hours (n=11). A clinical history design type is where the organisms clinical history of diagnosis, treatments, e.g. vaccinations, surgery etc. Time: lenght of co-culture Infection: whole-blood cells cultured ex-vivo in the presence of either uninfected red blood cells or red blood cells infected with Plasmodium falciparum Disease State: Healthy individuals with (PrevHxMal) or without (NoHxMal) previous exposure to complicated Plasmodium falciparum infection clinical_history_design
Project description:Gene methylation profiling of immortalized human mesenchymal stem cells comparing HPV E6/E7-transfected MSCs cells with human telomerase reverse transcriptase (hTERT)- and HPV E6/E7-transfected MSCs. hTERT may increase gene methylation in MSCs. Goal was to determine the effects of different transfected genes on global gene methylation in MSCs.
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.