Factors Associated With Emergency Department Health Professionals' Attitudes Toward Family Presence During Adult Resuscitation in 9 Greek Hospitals.

BACKGROUND: Different cultural context results in variability in health professionals' attitude toward family presence in the emergency department (ED) during resuscitation. Knowledge about emergency department health professionals' (EDHPs') attitude toward family presence during resuscitation (FPDR) in Greece can contribute to the understanding of cultural and value-related obstacles and opportunities to implement this practice. It would be useful for Greek health professionals when they have to care for citizens from other countries in which guidelines for FPDR exist to understand their difference in behavior and reaction and be more tolerant. OBJECTIVES: To explore Greek EDHPs' experiences, concerns, and benefits regarding FPDR and determine the contributing factors that are associated with their positive or negative attitude about the process. METHODS: A convenience sample of 305 health professionals working in the ED of 9 general hospitals of the National Healthcare System in northern Greece participated in a cross-sectional study (response rate, 87.6%). RESULTS: Positive experience of FPDR was encountered in 65.9% of EDHPs. Only 2.3% reported that they work in EDs where FPDR is allowed. Hospital regional location, previous positive experience of FPDR, and competency in supporting family members during FPDR were independently associated with a positive attitude toward FPDR. CONCLUSIONS: Greek EDHPs are not ready to accept FPDR practice. The establishment of formal policy similarly to resuscitation guidelines and family-centered training courses are needed to mitigate negative beliefs. Attempts to enhance positive attitude should include educational interventions providing knowledge on the benefits of FPDR.

Involvement of retrotransposon L1 in stemness and cellular plasticity.

Epithelial-to-mesenchymal transition (EMT) as well as the reverse process, mesenchymal-to-epithelial transition (MET) is important during embryogenesis. EMT is also involved in cancer invasion and metastasis, and can generate cells with properties similar to those of stem cells. Retrotransposons can rearrange the genome by inserting DNA in new loci, thus inducing mutations. This study examines the gene expression of transcription factors involved in EMT and MET. In the second experimental panel, the gene expression of L1 retrotransposon was studied. L1-open reading frame (ORF) 2 mRNA was found to be expressed both in cancer and cancer stem cells, while L1-ORF1 mRNA was expressed only in cancer cells. The suppression of L1-ORF2 gene expression demonstrated that this retrotransposon might affect EMT in colon cancer stem cells. This study highlights that the EMT process seems to differ between cancer cells and cancer stem cells, and that transposable elements seem to be involved in the process, influencing cellular plasticity.

Potential role for the Metnase transposase fusion gene in colon cancer through the regulation of key genes.

The Metnase fusion gene consists of a SET histone methyltransferase domain and a transposase domain from Mariner transposase. This transposable element is involved in chromosome decatenation, enhances DNA repair, promotes foreign DNA integration, and assists topoisomerase II function. This study investigates the role of Metnase in colon cancer homeostasis and maintenance of the stemness phenotype in colon cancer stem cells (CSCs). Silencing of Metnase was performed in human cancer cell lines before and after treatment with cisplatin, and in colon CSCs. Subsequent changes in the expression of genes involved in repair mechanisms, DNA synthesis, topoisomerase II function, and metastasis as well stemness transcription factors were studied with RT-qPCR experiments. Cellular viability and apoptosis were evaluated by flow cytometry. The results suggest that Metnase influences the expression of many genes involved in the above processes. Furthermore, Metnase levels appear to impact upon expression of NANOG, OCT3/4, and SOX2. Suppression of Metnase also led to an increase in apoptosis. Therefore, Metnase may possess an important role in DNA repair, topoisomerase II function, and the maintenance of stemness during colon cancer development.

Use of the comet assay technique for quick and reliable prediction of in vitro response to chemotherapeutics in breast and colon cancer.

BACKGROUND: Determination of response to chemotherapy is a major requirement of personalized medicine. Resistance, whether developed or native, critically affects a treatment's success. Single Cell Gel lectrophoresis - also known as a comet assay - is used to detect DNA damage at the level of individual eukaryotic cells. We assessed the use of comet assays in determining response to chemotherapeutic drugs that are widely used in breast and colon cancer. RESULTS: We treated human breast and colon cancer cell lines with melphalan, cisplatin, mechlorethamine or doxorubicin, as monotherapies. Drug activities varied even in the same cancer types, further demonstrating the heterogeneity of different cancer types. CONCLUSION: The comet assay technique can provide reliable and quick results with minimum requirements and is applicable to a wide variety of drugs.

Study of the interaction among Notch pathway receptors, correlation with stemness, as well as their interaction with CD44, dipeptidyl peptidase-IV, hepatocyte growth factor receptor and the SETMAR transferase, in colon cancer stem cells.

CONTEXT: The Notch signaling pathway is one of the most important pathways during normal development and implicated in self-renewal of adult stem cells and differentiation of progenitor cells. Abnormal expression of Notch receptors has been associated with many epithelial metaplastic and neoplastic lesions. OBJECTIVE-MATERIALS AND METHODS: In this particular study, it was determined the relative gene expression of Notch receptors after knockdown experiments in colon cancer stem cells (CSCs) and the gene expression changes in stemness transcription factors (Oct4, Sox2, Nanog), as well in dipeptidylpeptidase-4, CD44 antigen, Met proto-oncogene and in Metnase transposase. RESULTS: In control CSCs Notch-2 had the higher expression, followed by Notch-1, Notch-3. Notch-4 demonstrated the lower gene expression among the receptors. The suppression of Notch-1 led to increased expression of Oct4 and Sox2, but in decreased gene expression of cMET, Setmar and CD44. The CD26 expression remained unchanged. The knockdown of Notch-2 led to decreased expression of all transcription factors. Notch-3 down regulation caused increased Oct4 gene expression, but decreased levels for the rest of the genes. Finally, the suppression of Notch-4 had the same effect as in receptor Notch-3. DISCUSSION AND CONCLUSION: The above experimental data suggest the possible interaction between the four different receptors of Notch signaling pathway. The expression of CD26, cMET and N-methyltransferase Setmar was also changed. Finally, the stemness phenotype was changed in a different way each time, according to the receptor that was down regulated. All Notch receptors and particularly Notch-2 seem to play an important role in cancer stem cells.

AP-1 Gene Expression Levels May Be Correlated with Changes in Gene Expression of Some Stemness Factors in Colon Carcinomas.

The AP-1 transcription factor is a heterodimer protein that regulates gene expression in response to a variety of extrinsic stimuli through signal transduction. It is involved in processes including differentiation, proliferation, and apoptosis. Among the genes it regulates are transcription factors that contribute to the stemness phenotype. Cancer stem cells have the ability to self-renew and initiate differentiation into heterogenic cancer cells, which may cause metastasis and relapses. In the present study, we evaluated the effect of AP-1 complexes, as well as the C-FOS and C-JUN genes, in relation to NANOG, OCT3/4, and SOX2 transcription factors. All assays were undertaken with colon cancer stem cells. Knockdown experiments with siRNA were performed for each individual gene as well as their combination. Changes in gene expression were calculated with quantitative polymerase chain reaction experiments, while the effect on cell cycle distribution and apoptosis was studied by flow cytometry. The results differed depending on the percentage of repression, as well as the gene that was suppressed. In all cases, the number of apoptotic cells was increased. These findings indicate that AP-1 may have a crucial role in the maintenance of cancer stem cells.