We identified conditions under which Buffalo green monkey cells grew on the surfaces of cellulose nitrate membrane filters in such a way that they covered the entire surface of each filter and penetrated through the pores. When such conditions were used, poliovirus that had previously been adsorbed on the membranes infected the cells and replicated. A plaque assay method and a quantal method (most probable number of cytopathic units) were used to detect and count the viruses adsorbed on the membrane filters. Polioviruses in aqueous suspensions were then concentrated by adsorption to cellulose membrane filters and were subsequently counted without elution, a step which is necessary when the commonly used methods are employed. The pore size of the membrane filter, the sample contents, and the sample volume were optimized for tap water, seawater, and a 0.25 M glycine buffer solution. The numbers of viruses recovered under the optimized conditions were more than 50% greater than the numbers counted by the standard plaque assay. When ceftazidime was added to the assay medium in addition to the antibiotics which are typically used, the method could be used to study natural samples with low and intermediate levels of microbial pollution without decontamination of the samples. This methodological approach also allowed plaque hybridization either directly on cellulose nitrate membranes or on Hybond N؉ membranes after the preparations were transferred.

BackgroundPlatinum derivatives are used widely for the treatment of many cancers. However, the toxicity that is observed makes imperative the need for new drugs, or new combinations. Anvirzel™ is an extract which has been demonstrated with experimental data that displays anticancer activity. The aim of the present study is to determine whether the combination of Cisplatin and Anvirzel™ has a synergistic effect against different types of cancer.Materials and methodsTo measure the efficacy of treatment with Cisplatin and Anvirzel™, methyl-tetrazolium dye (MTT) chemosensitivity assays were used incorporating established human cancer cell lines. Measurements were performed in triplicates, three times, using different incubation times and different concentrations of the two formulations in combination or on their own. t-test was used for statistical analysis.ResultsIn the majority of the cell lines tested, lower concentrations of Anvirzel™ induced a synergistic effect when combined with low concentrations of Cisplatin after an incubation period of 48 to 72 h. The combination of Anvirzel™/Cisplatin showed anti-proliferative effects against a wide range of tumours.ConclusionThe results showed that the combination of Anvirzel™ and Cisplatin is more effective than monotherapy, even when administered at low concentrations; thus, undesirable toxic effects can be avoided.

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.

Cancer stem cell-like cells (CSCs) are cancer cells that have the ability of self-renewal and differentiation into multiple malignant cell types (hierarchy). Thus, can cause relapses and metastasis. CSCs' phenotype is defined by special transcription factors such as Nanog, Oct3/4, Sox2, Nestin, and CD34. The present study aims to determine the change in gene expression of the above markers in correlation with the stage of the disease in breast cancer patients. Initially, whole blood samples from patients with breast cancer were collected, followed by the isolation and culture of circulating tumor cells (CTCs). This was followed by the quantification of CSCs from the above cultures. CSCs' molecular analysis was performed with qPCR, with the use of gene specific primers. At the same time of the analysis, the clinical assessments of the patients were requested from their physicians. The results indicated a linear relationship between the gene expression of stemness markers and the stage of the disease, as well as specific expression patterns by stage. It seems that these genes have an important role in the progression of the disease, thus they might be target for new treatment approaches.

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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.

A fundamental problem in cancer research is identification of the cells responsible for tumor formation. The latest field of cancer research has revealed the existence and role of cancer stem cells (CSCs). These findings support the idea that malignancies originate from a small fraction of cancer cells that show self-renewal and multi- or pluripotency. Identification of this CSC population has important implications for the management of cancer patients, including diagnostic and predictive laboratory assays as well as novel therapeutic strategies that specifically target CSCs. In this study, we investigated the growth rates of CSC populations for comparison with cancer cell lines. To construct the growth curves, blood-derived CSCs were isolated from patients with breast, colon, or lung cancer and cultured in vitro. Quantitative real-time PCR was then performed to identify CSCs in the samples. We found that CSCs did not follow the common pattern of a typical growth curve of mammalian cells in contrast to the cancer cell lines. This observation of rapidly growing CSCs indicates their involvement in tumor formation.

The aim of the paper is to examine the uses of CALL among students and teachers in Cyprus private secondary education. The notions of digital native and digital immigrant are revisited as part of the framework set for understanding the current situation in CALL exploitation and future potential within and beyond the classroom walls. A survey investigation is conducted for the collection of the data. Results indicate that a third category of users emerges, the 'inbetweeners' who may have a key role to play in CALL integration.