Project description:Glutamicibacter arilaitensis grown in large scale liquid Cheese Curd media and extracted with Amberlite XAD16, back extracted with MeOH:DCM 50:50, separated on SPE column to afford 7 fractions labeled A through G based on increasing polarity (A = 10% MeOH, F = 100% MeOH and G = 100% EtoAC. Fractions E(60% MeOH), D(80% MeOH), and F(100% MeOH) run on a qTOF and submitted here. suffix 1 refers to treatment of cultures with desferrioxamine and suffix PLAIN means untreated. JB182new_20170614 is a crude extract from a subsequent small scale growup without desferrioxamine.
Project description:We investigated the effects of the crude extract of a South African medicinal plant, Cotyledon orbiculata, on cell survival of colon (HCT116) cancer cell lines. Using RNASeq, we discovered that the extract interfered with mRNA regulatory pathways. Here, we found that the extract of Cotyledon orbiculata, a South African medicinal plant, had an anti-proliferative effect in cancer cells, mediated by apoptosis induced by alternative splicing of hnRNPA2B1 and BCL2L1.
Project description:In order to get mechanistic insights into the cardiomyogenic and differentiation-promoting activity of Crataegus spp. Extract WS®1442 and the identified bioactive subfractions (MeOH eluate), we have employed whole genome microarray expression profiling after 6 hours and 24 hours of treatment. We found that stress-associated genes are affected as well as specific signaling pathways such as TGFbeta, FGF, BDNF and retinoic acid.
Project description:Untargeted metabolomics (LC-MS/MS) of water samples (SPE on Strata XL Phenomonex) and of macroalgal whole tissues (50% MeOH/DCM) and surface extracts (MeOH). Positive and negative ionization mode
Project description:One of the central goals for the development of cell-free (CF) gene expression systems is to ensure reproducible expression of proteins at high quality and yield. Over the past decades, starting from crude cell extracts, a variety of successful preparation protocols have been developed and optimized reaction conditions have been established. One of the crucial steps during the preparation of cell extract-based expression systems, however, is the cell lysis procedure itself, which largely determines the quality of the active components of the extract. Here, we demonstrate for an E. coli based system that a lysis procedure combining incubation of the cells with lysozyme with a gentle sonication step results in highly active cell extracts. As examples for the capabilities of our extract, we demonstrate the production of several fluorescent proteins as well as the production and assembly of T7 bacteriophages. Compared to other cell-free expression systems, our method achieved the highest phage titers in our plaque assays. Stateof-the-art quantitative proteomics revealed that enzymes of the energy regeneration pathway were enriched in our extract compared to a widely used commercial cell extract. On the other hand, ribosomal proteins were found to be more abundant in the commercial product.
Project description:Bone marrow stromal cells (BMSCs) are multipotent stem cells that preferentially differentiate into mesenchymal cells. If they can be dedifferentiated into embryonic stem cell-like cells, they will be a highly attractive source for cell therapy. Cell and egg extracts have been used in a few studies to evaluate nuclear reprogramming, but these have not examined cell pluripotency in any detail. In this study, we used a cell reversible permeabilization method to treat BMSC with Xenopus laevis mitotic egg extract. We observed an upregulation of the pluripotent protein Oct3/4 in BMSCs treated by this extract. We also further evaluated transcriptional changes with a focused stem cell oligonucleotide array. A number of genes involved in the Notch or Wnt signaling pathways were upregulated in BMSC exposed to Xenopus egg extract. In conclusion, our microarray data from BMSCs exposure to egg extracts may provide interesting clues regarding factors involved in nuclear reprogramming. Our approach is an alternative method towards dedifferentiation of cells without genetic modification, which is preferable in the clinical situation. Keywords: Cell type comparison
Project description:To assess the influences of a crude medicinal herb extract (MHE) on the immune composition and function in inguinal white adipose tissue (iWAT), we isolated iWAT stromal vascular fractions (SVFs) from control (PBS) and MHE-treated mice and performed RNA-seq analysis. iWAT SVF of mice treated with MHE was defined as the treatment group. Phosphate-buffered saline (PBS) treatment was used as the control group. Then RNA-Seq experiment was performed by OE Biotech Co., Ltd. (Shanghai, China) to analyze gene expression changes in iWAT SVF.