Development of a model for the two major types of prostate carcinomas that enables the detection and study of potential markers for predicting metastasis into lymph nodes

Prostate cancer is the one of the most frequent types of cancer in males. Until today the prostate carcinoma cases were evaluated for the risk of metastases on criteria like the scoring of D Amico or Candiolo but without the possibility to predict the hematogenic or lymphatic route of metastases. In this study three human prostate cancer cells lines which are hormone sensitive have been selected (LNCaP, VcaP and MDAPCa2b) and three which are hormone refractory (PC3, DU145 and SerBob). Then each line was forming co-culture with endothelial cell or osteoblast in order to form tumoroids and resemble the lymphatic or bone metastases respectively. Then a differential expression analysis of all transcriptomes took place, which reveal several genes that are steadily overexpressed during co-cultivation. Then knock down methodologies reveal that only few genes and relevant proteins are involved in the metastatic process especially to the lymph nodes (MYO1B, MYADM and PIP5K1A). These potential biomarkers also validated by comparing prostate carcinoma cases and healthy individuals. Of course, the actual proof of concept requires further in vivo studies in orthotopic animal models, but nevertheless there is a strong indication that these theses biomarkers may have a value to predict the metastases of a prostate carcinoma to the lymph nodes. This may provide more stronger indication to the urologist to safely select which patient may have an actual clinical benefit from an inguinal lymph node clearance which not

QUALITY ASSESSMENT AND QUANTIFICATION OF GENISTEIN IN DIETARY SUPPLEMENTS BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY, QUANTITATIVE NUCLEAR MAGNETIC RESONANCE, AND TWO-DIMENSIONAL DIFFUSION ORDERED SPECTROSCOPY 1H

Objective: The aim of the present study is to analyze four herbal dietary supplements containing genistein by liquid chromatography, quantitative nuclear magnetic resonance (qNMR), and diffusion ordered spectroscopy (DOSY) 1H NMR. Methods: Quantification of the active ingredient, genistein, is carried out by high-performance liquid chromatography (HPLC) and qNMR. Two-dimensional (2D) DOSY NMR also allows the qualitative analysis of the samples with the detection of active ingredient and excipients present in the formulations. Results: The validated HPLC and qNMR methods showed that all four supplements contain genistein in different amounts, and 2D DOSY NMR provides a clear image of all ingredients in the formulations. Conclusion: The use of the three techniques provides detailed information on each product and its contents, and all of them are currently used for the quality control of natural supplements by our laboratory

Bacterial and fungal microflora in surgically removed lung cancer samples

<p>Abstract</p> <p>Background</p> <p>Clinical and experimental data suggest an association between the presence of bacterial and/or fungal infection and the development of different types of cancer, independently of chemotherapy-induced leukopenia. This has also been postulated for the development of lung cancer, however the prevalence and the exact species of the bacteria and fungi implicated, have not yet been described.</p> <p>Aim</p> <p>To determine the presence of bacterial and fungal microflora in surgically extracted samples of patients with lung cancer.</p> <p>Materials and methods</p> <p>In this single-center prospective, observational study, tissue samples were surgically extracted from 32 consecutive patients with lung cancer, and reverse-transcription polymerase chain reaction (RT-PCR) was used to identify the presence of bacteria and fungi strains.</p> <p>Results</p> <p>The analysis of the electrophoresis data pointed out diversity between the samples and the strains that were identified. Mycoplasma strains were identified in all samples. Strains that appeared more often were Staphylococcus epidermidis, Streptococcus mitis and Bacillus strains, followed in descending frequency by Chlamydia, Candida, Listeria, and Haemophilus influenza. In individual patients Legionella pneumophila and Candida tropicalis were detected.</p> <p>Conclusions</p> <p>A diversity of pathogens could be identified in surgically extracted tissue samples of patients with lung cancer, with mycoplasma strains being present in all samples. These results point to an etiologic role for chronic infection in lung carcinogenesis. Confirmation of these observations and additional studies are needed to further characterize the etiologic role of inflammation in lung carcinogenesis.</p

9-(4-Methoxyquinazolin-2-yl)-9H-purin-6-amine

A novel hybrid consisting of quinazoline and adenine moieties has been synthesized as a precursor of a potential biologically active target compound. The structure of 9-(4-methoxyquinazolin-2-yl)-9H-purin-6-amine (2) was characterized and confirmed using the following spectroscopic methods: LC-UV-MS, 1H-NMR, 13C-NMR and HSQC-NMR

Evaluation of a simple method for storage of blood samples that enables isolation of circulating tumor cells 96 h after sample collection

Abstract Background Minimizing the effects of transportation on the properties of biological material is a major challenge for the scientific community. The viability of cells is important in cases where their study is urgent for evaluation of treatment response or for the study of cancer progression. Circulating tumor cells (CTCs) constitute a cell subpopulation with great importance for oncologists, because of their prognostic value. Detection and isolation of CTCs from blood samples is a routine activity in many laboratories, but concerns exist with regard to the maintenance of the cells during transportation. In this study, experiments were conducted to determine the stability of gene and protein expression in CTCs over a period of 96 h. Results Blood samples collected from healthy individuals and patients with cancer were each divided into five aliquots, which were stored at 2–8 °C and analyzed after 0, 24, 48, 72 and 96 h of storage. CTCs from patients and CD45-negative cells from healthy individuals were isolated each day using enrichment protocols, and qPCR was performed to determine expression levels of genes encoding specific biological markers. In addition, cells from breast and colon cancer cell lines were spiked into blood samples from healthy individuals, and these samples were stored and analyzed over a period of 96 h by PCR and by flow cytometry. The markers that were studied included housekeeping genes and genes associated with the response to chemotherapy, as well as genes encoding transcription factors. The results demonstrated that the expression profiles of specific genes and proteins in CTCs were not significantly affected by 72 h of storage. After 96 h of storage, expression of some genes was altered. Conclusion The transportation of blood at low temperature (2–8 °C) in the presence of the anticoagulant EDTA can protect CTCs from alteration of gene and protein expression for at least 72 h. Furthermore, under these conditions, CTCs can be detected and isolated 96 h after blood collection

Evaluation of Tegaran Formula ZhenHua cytotoxicity against human cancer cell lines.

The aim of this study is to evaluate the potential health effects of Tegaran Formula ZhenHua, a nutritional supplement used mainly by cancer patients. Its active ingredients and cytotoxicity was assessed with analytical methods and viability assays, respectively. The analytical methods consisted of dissolution, disintegration, HPLC, LC/MS, GC/MS and NMR. Cytotoxicity was assessed by MTT, SRB, CVE colorimetric viability assays in 0, 24, 48 and 72h time points. The results indicate that Tegaran Formula ZhenHua supplement did not present any cytotoxic effects due to issues related to the capsules' solubility, distribution and identification of the active ingredient