Association of self-DNA mediated TLR9-related gene-, DNA methyltransferase and cytokeratin protein expression alterations in HT29-cells to DNA fragment length and methylation status

To understand the biologic role of self-DNA bound to Toll-like Receptor 9 (TLR9), we assayed its effect on gene and methyltransferase expressions and cell differentiation in HT29 cells. HT29 cells were incubated separately with type-1 (normally methylated/nonfragmented), type-2 (normally methylated/fragmented), type-3 (hypermethylated/nonfragmented), or type-4 (hypermethylated/fragmented) self-DNAs. Expression levels of TLR9-signaling and proinflammatory cytokine-related genes were assayed by qRT-PCR. Methyltransferase activity and cell differentiation were examined by using DNA methyltransferase (DNMT1, -3A, -3B) and cytokeratin (CK) antibodies. Treatment with type-1 DNA resulted in significant increase in TLR9 expression. Type-2 treatment resulted in the overexpression of TLR9-related signaling molecules (MYD88A, TRAF6) and the IL8 gene. In the case of type-3 treatment, significant overexpression of NFkB, IRAK2, and IL8 as well as downregulation of TRAF6 was detected. Using type-4 DNA, TRAF6 and MYD88A gene expression was upregulated, while MYD88B, IRAK2, IL8, and TNFSF10 were all underexpressed. CK expression was significantly higher only after type-1 DNA treatment. DNMT3A expression could also be induced by type-1 DNA treatment. DNA structure may play a significant role in activation of the TLR9-dependent and even independent proinflammatory pathways. There may be a molecular link between TLR9 signaling and DNMT3A. The mode of self-DNA treatment may influence HT29 cell differentiation

Stability and Flexibility in Psychotherapy Process Predict Outcome

Ten good outcome and ten poor outcome psychotherapy cases were compared to investigate whether or not the temporal stability and flexibility of their process variables can predict their outcomes. Each participant was monitored daily using the Therapy Process Questionnaire (TPQ), which has 43 items and seven sub-scales, and responses over time were analyzed in terms of correlation robustness and correlation variability across the TPQ sub-scales. “Correlation robustness” and “correlation variability” are two basic characteristics of any correlation matrix: the first is calculated as the sum of the absolute values of Pearson correlation coefficients, the second as the standard deviation of Pearson correlation coefficients. The results demonstrated that the patients within the poor outcome group had lower values on both variables, suggesting lower stability and flexibility. Furthermore, a higher number of cycles of increase and decrease in correlation robustness and variability of the TPQ sub-scales was observed within good outcome psychotherapies, suggesting that, these cycles can be considered as process-markers of good-outcomes. These results provide support for the validity of these quantitative process-parameters, correlation robustness and variability, in predicting psychotherapeutic outcomes. Moreover, the results lend support to the common clinical experience of alternating periods of flexibility and integration being beneficial to good psychotherapeutic processes

Cell Free DNA of Tumor Origin Induces a 'Metastatic' Expression Profile in HT-29 Cancer Cell Line

BACKGROUND: Epithelial cells in malignant conditions release DNA into the extracellular compartment. Cell free DNA of tumor origin may act as a ligand of DNA sensing mechanisms and mediate changes in epithelial-stromal interactions. AIMS: To evaluate and compare the potential autocrine and paracrine regulatory effect of normal and malignant epithelial cell-related DNA on TLR9 and STING mediated pathways in HT-29 human colorectal adenocarcinoma cells and normal fibroblasts. MATERIALS AND METHODS: DNA isolated from normal and tumorous colonic epithelia of fresh frozen surgically removed tissue samples was used for 24 and 6 hour treatment of HT-29 colon carcinoma and HDF-alpha fibroblast cells. Whole genome mRNA expression analysis and qRT-PCR was performed for the elements/members of TLR9 signaling pathway. Immunocytochemistry was performed for epithelial markers (i.e. CK20 and E-cadherin), DNA methyltransferase 3a (DNMT3a) and NFkappaB (for treated HDFalpha cells). RESULTS: Administration of tumor derived DNA on HT29 cells resulted in significant (p/=1, p/=1, p</=0.05), including increased expression of key adaptor molecules of TLR9 pathway (e.g. MYD88, IRAK2, NFkappaB, IL8, IL-1beta), STING pathway (ADAR, IRF7, CXCL10, CASP1) and the FGF2 gene. CONCLUSIONS: DNA from tumorous colon epithelium, but not from the normal epithelial cells acts as a pro-metastatic factor to HT-29 cells through the overexpression of pro-metastatic genes through TLR9/MYD88 independent pathway. In contrast, DNA derived from healthy colonic epithelium induced TLR9 and STING signaling pathway in normal fibroblasts

Growing up -the completion of the VLTI

Abstract. The completed VLTI with eight Delay Lines and eight ATs forms the basis for the second generation instrumentation. We describe the events up to first fringes with the test instrument VINCI using the siderostats, and the planning for the immediate future. Multi beam combination for &apos;smoother images&apos; will be briefly discussed as well as artificial guide stars for fringe tracking. New technological developments like fiber optics amplifiers and integrated optics in combination with STJ open the door for a new type of interferometric arrays. Baselines as long as a a few kilometres come into reach. Examples of these second generation interferometers will be given

Poupart-szalag rekonstrukciója autológ fascia lata lebennyel. Az első ismertetett hazai eset

INTRODUCTION: A technique of reconstructing the inguinal ligament using pedicled fascia lata flap is described. PRESENTATION OF CASE: A 66-year-old woman was referred with massive incarcerated left inguinal hernia, following acute surgery on a femoral vein leasion and numerous attempts at repair and subsequent recurrences. There was complete absence of the left inguinal ligament. The inguinal ligament was reconstructed using a strip of fascia lata, pedicled on the anterior superior iliac spine. This was transposed to cover the external iliac vessels, and sutured to the pubic tubercle. The musculoaponeurotic abdominal wall was reconstructed with 15x13 cm sheet of polypropylene mesh, placed preperitoneal and sutured to the remaining abdominal wall muscles and to the neo-Pouoart ligament. DISCUSSION: Complete destruction of the inguinal ligament is rare but can occur following multiple operative procedures or trauma. Published reports of inguinal ligament reconstruction have been performed using synthetic mesh. The use of autologous tissue should reduce the risk of erosion into the neurovascular bundle, seroma formation, and enhance integration into surrounding tissues. CONCLUSION: This new technique for autologous reconstruction of the inguinal ligament provides a safe alternative to the use of synthetic mesh in the operative armamentarium of plastic and general surgeons. This is the first reported case in Hungary

Role of DNA Methylation in Colorectal Carcinogenesis

Colorectal cancer is the most common malignancy of the gastrointestinal tract and a leading cause of cancer-related deaths worldwide. In order to detect early precursor lesions, colonoscopy is widely used. Unfortunately, patient adherence to colonoscopy is poor, which is partially due to the modest performance of currently used prescreening tests. Recently, epigenetics added an additional layer to the understanding of colorectal carcinogenesis. DNA methylation as part of the epigenetic gene-silencing complex is a universally occurring change in colorectal cancer and arises prior to the onset of recognizable preneoplastic changes, which may have huge preventive implications. Herein we discuss the major developments in the field of colorectal carcinogenesis and DNA methylation, including alterations in nonneoplastic conditions such as aging and ulcerative colitis. We try to demonstrate how this epigenetic modification can be harnessed to address some of the key issues impeding the successful clinical management of colorectal cancer. Copyright (C) 2012 S. Karger AG, Base

Neuroprotektion: Von bench -Zelluläre Mechanismen- und von bedside -Moderne Schlaganfallakuttherapie

<p>Number of living(A) and dead(B) HT-29 cells determined by Trypan blue exclusion test; PI cell-cycle analysis of HT-29 colon cancer cells(C).</p

Table representing RNA expression change of CDH1 and KRT 20 after normal and tumor tissue derived DNA treatment.

<p>Table representing RNA expression change of CDH1 and KRT 20 after normal and tumor tissue derived DNA treatment.</p

Significant gene expression changes after tumor tissue-originated DNA treatment in HT-29 cells.

<p>Significant gene expression changes after tumor tissue-originated DNA treatment in HT-29 cells.</p