ABSTRACT: In patients with primary breast cancer, neoadjuvant chemotherapy with doxorubicin plus pemetrexed followed by docetaxel (AP-D) is associated with a pathologic complete response (pCR) rate of 16.5%, and doxorubicin plus cyclophosphamide followed by docetaxel (AC-D) is associated with a pCR rate of 20.2%. Our primary objective was to identify single predictive genetic markers for achievement of pCR following either AP-D or AC-D treatment. Our main secondary objective was to detect treatment-group specific, pCR-predictive gene signatures.
Project description:Effect of PAR and temperature stress on the gene expression Saccharina latissima. Total RNA of stress treatments (low PAR 2° and 17°C, high PAR 2° and 17°C) was hybridized against the control treatment (low PAR 12°C); hybridizations were carried out in 4 replicates.
Project description:Streptococcus mutans was grown for 48 h in a biofilm in the absence (single species) and in the presence (dual species) of Veillonella parvula. In addition V. parvula single species 48 h biofilms were grown, to be used as a control. RNA was harvested from all types of biofilms and the transcript levels of the two types of biofilms containing S. mutans were compared with the use of S. mutans microarrays. V. parvula RNA was hybridized to S. mutans microarrays as a control for possible cross-hybridisation.
Project description:Helicobacter pylori are gram-negative bacteria that colonize the human stomach and are the major etiological factor in gastric carcinoma development. The aim of this work was to evaluate changes in gene expression in gastric cells induced by H. pylori. The human gastric carcinoma-derived cell line AGS was infected with H. pylori strain 60190 (ATCC 49503) for 24 hours. RNA was extracted from three independent experiments.
Project description:A method based on a modified broad-range PCR and an oligonucleotide microarray for the simultaneous detection and identification of 12 bacterial pathogens at the species level. The feasibility of this assay in routine diagnostic testing was evaluated using 146 blood culture positive and 40 blood culture negative samples. The processed data are composed of ORGANISM NAME or NEGATIVE answer depending if the pathogen is detected in the sample or not. See supplementary file linked below.
Project description:Human skeletal myoblast cell line LHCN-M2 (Zhu et al., Aging Cell (2007), vol. 6, pp 515-523) were plated on collagen-coated 6-well Falcon cell culture plates at 500 000 cells/well, differentiation was induced 24h later by switching to DMEM supplemented with 0,01 mg/ml insulin and 0,1 mg/ml transferrin. Cells were collected at 0, 3, and 7 days of differentiation time points. Total RNA was extracted and miRNA expression evaluated by quantitative real-time PCR. MiRNA expression profiling in proliferating and differentiating (3 and 7 days) human skeletal myoblasts LHCN-M2.
Project description:We describe mass spectrometric antibody-peptide binding motif analyses of a monoclonal antibody (clone 18E9.D9, BioLegend, London, England) from mouse which was raised against the predominantly occuring phosphorylated linker sequence HpTGEKP of human C2H2 zinc finger proteins. As the anti-HpTGEKP antibody is assumed to bind to related phosphorylated C2H2 ZNF protein linker sequences as well, specific amino acid exchanges were investigated for evaluating their importance for immune complex formation. Two point mutations of the zinc finger protein 331 (ZNF331) gene, causing single amino acid exchanges R9I and H11Y, respectively, have been found to be significantly enriched in Uterine Corpus Endometrial Carcinoma, Colon and Rectal Adenocarcinomas, and Skin Cutaneous Melanoma. Since the H11Y amino acid exchange affects histidine residue H1 of the HpTGEKP linker motif which is recognized by the anti-HpTGEKP antibody, we examined in this work whether the in several cancers mutated linker amino acid sequence YTGEKP and its phosphorylated derivatives YpTGEKP, pYTGEKP, and pYpTGEKP would be bound by the anti-HpTGEKP antibody. Immune complex formation ability was compared to that of peptides HTGEKP, HpTGEKP, and HpTGKKP, respectively, which served as controls. Binding behaviors towards the anti-HpTGEKP antibody were investigated by "Intact Transmission Epitope Mapping - Thermodynamic Weak-force Order (ITEM-TWO)".
Project description:Mesenchymal stem cells (MSC) resemble a multipotent adult stem cell population capable of differentiation into a number of different mesodermal cell types including adipodytes, osteoblasts, chondroblasts. Although still in debate there is some evidence, that these cells can also differentiate into cells of non-mesodermal germal layers including hepatocytes, myocytes, cardiomyocytes or neurons. Analysis of these cells in the course of differentiation makes them an intriguing model for the examination of stemness. More importantly the differentiation capacity of MSC raises high hopes for clinical applications. In this study we have isolated MSC from bone marrow under two different culture conditions, from adipose tissue and from umbilical cord blood. The genome wide gene expression profile of these different human MSC was compared with reference RNA of non-multipotent human fibroblasts HS68. The aim of this study was to elucidate common molecular characteristics of MSC as well as differences in their expression profile. These results might help for a better understanding of MSC and contribute to a reliable quality control that will be necessary for clinical applications.
Project description:Plasma cells (PCs) as effectors of humoral immunity produce immunoglobulins to match pathogenic insult. However, emerging data suggests more diverse roles for PCs as regulators of immune and inflammatory responses via secretion of factors other than immunoglobulins. The extent to which such responses are pre-programmed in B-lineage cells or can be induced in PCs by the microenvironment is unknown. Here we dissect the impact of IFNs on the regulatory networks of human plasma cells. We show that core PC programs are unaffected, while PCs respond to IFNs with distinctive transcriptional responses. The ISG15-system emerges as a major transcriptional output induced in a sustained fashion by IFN-α in PCs and linked both to intracellular conjugation and ISG15 secretion. This leads to the identification of ISG15-secreting plasmablasts/PCs in patients with active SLE. Thus ISG15-secreting PCs represent a distinct pro-inflammatory PC subset providing an immunoglobulin-independent mechanism of PC action in human autoimmunity B-cells were isolated from the peripheral blood of three adult donors and differentiated in vitro (see individual samples for culture conditions)
Project description:Bacillus thuringiensis, a well-known and effective bio-insecticide, has attracted considerable attention as a potential biological control agent for the suppression of plant diseases. Treatment of tomato roots with a filter-sterilized cell-free filtrate (CF) of B. thuringiensis systemically suppresses bacterial wilt caused by Ralstonia solanacearum through systemic activation of the plant defense system. Comparative analysis of the expression of the Pathogenesis-Related 1(P6) [PR-1(P6)] gene, a marker for induced resistance to pathogens, in various tissues of tomato plants treated with CF on their roots suggested that the B. thuringiensis-induced defense system was activated in the leaf, stem, and main root tissues, but not in the lateral root tissue. At the same time, the growth of R. solanacearum was significantly suppressed in the CF-treated main root tissue but not in the CF-treated lateral root tissue. This distinct activation of the defense reaction and suppression of R. solanacearum were reflected by the differences in the transcriptional profiles of the main and lateral tissues in response to the CF. In the CF-treated main root tissue, but not CF-treated lateral root tissue, the expression of several salicylic acid (SA)-responsive defense-related genes was specifically induced, whereas jasmonic acid (JA)-related gene expression was either down-regulated or not induced in response to the CF. On the other hand, genes encoding ethylene (ET)-related proteins were induced equally in both the main and lateral root tissues. Taken together, the co-activation of SA-dependent signaling pathway with ET-dependent signaling pathway and suppression of JA-dependent signaling pathway may play key roles in B. thuringiensis-induced resistance to R. solanacearum in tomato plants. Gene expression was measured in main and lateral root tissues of tomato treated with Bacillus thuringiensis or distilled water-treated control at 48 hours after treatment. Two independent experiments were performed at each tissue (main root or lateral root tissue) for each treatment (Bacillus thuringiensis or distilled water control).