Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) settingg

Aim: To compare our parameters as regards: i) cell count via two different automated cell count techniques, and ii) viability via automated trypan blue exclusion and 7-aminoactinomycin D (7-AAD) staining. Method: We used the trypan blue exclusion technique and an automated cell counter and for viability testing, and the trypan blue exclusion technique and the 7-AAD evaluation by flow cytometry. The trypan blue exclusion and the radio frequency techniques were used for automated cell counting. Flow cytometric analysis was performed by evaluating the yielded cellular products for 7-AAD uptake during the cell count of CD34+ cells. Results: The mean values for cell count were estimated as 3.44±1.22x106/ml (range, 2.48-5.71x106/ml) and 4.14±1.94x106/ml (range, 1.77-7.43x106/ml) for the trypan blue exclusion and radio frequency techniques, respectively. Additionally, the mean values for viability analyses via the automated trypan blue exclusion and 7-AAD were 93.38±6.09% (range, 79.00-98.00%) and 99.49±0.60% (range, 98.40-100.00%), respectively. Conclusions: Our study has responded to two fundamental questions: whether the results of both of the automated techniques for cell count correspond with each other, and whether the results of the automated viability assessment conform those of the 7-AAD technique during the manufacturing processes of cellular therapy products intended for clinical use. Even though we have the opportunity to use the hemocytometer in our laboratory setting, the automated trypan blue exclusion technique gives cell count results in concordance within the range of the expectations of our Quality Management System (QMS)

A Preliminary Investigation on the Presence of Calcifying Nanoparticles in the Breast Tumor

Calcium phosphate is deposited in many diseases, but the molecular basis of mineralization remains largely unknown. Biomineralizied calcifications that are formed by calcium deposits are also detected in breast mammograms. Some of the detected microcalcifications are thought to be related with malignancy. Taken together, calcifying nanoparticles (CNP) may be thought as a source of malign calcifications in breast cancers. The aim of the study is to research the presence of CNP in breast tumor tissue. With this aim, the presence of CNP was investigated by culturing 16 patients' breast tumor tissue and from 2 pathologic tissues with transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Their growth was monitored by optical density (OD) at a wavelength of 650 nm. CNP couldn't be found in the analysed tissues. The presence of CNP in the breast tumor tissue was researched for the first time. We could not find CNP in the breast tumor tissue, but we think this research will open a new field of study for researchers

Cost analysis and characteristics of the patients admitted to emergency service with poisoning

Objective: To investigate the cost analysis and hospitalization rates and modes of poisoning of patients who applied to Bakent University Ankara Hospital Adult Emergency Service. Methods: Poisoning Patients who applied to emergency service due to suicide attempt between 01.01.2011-31.12.2014 were included in the study. Patients' age, gender, date of admission, definite diagnosis, the rate of hospitalization, and hospital costs were examined. A total of 646 patients were included in the study. 208 (32.2%) of the patients were male. The median age of the patients was 29 years. Results: The most causes of poisoning was drug intake. The median cost of the patients was 75.14 TL (IQR: 66.5). Conclusion: There was a positive correlation between age and cost (P<0.05). The majority of the patients apply to emergency dapartment with drug poisoning and female

Genetic engineering of hematopoiesis for targeted IFN-α delivery inhibits breast cancer progression

The immunosuppressive tumor microenvironment represents a major hurdle to cancer therapy. We developed a gene transfer strategy into hematopoietic stem cells (HSCs) to target transgene expression to tumor-infiltrating monocytes/macrophages. Using a combination of transcriptional and microRNA-mediated control, we achieved selective expression of an interferon-α (IFN-α) transgene in differentiated monocytes of human hematochimeric mice. We show that IFN-α transgene expression does not impair engraftment and long-term multilineage repopulation of NSG (NOD/LtSz-scidIL2Rγ) mice by transplanted human HSCs. By providing a source of human cytokines in the mice, we improved the functional reconstitution of human myeloid, natural killer, and T cell lineages, and achieved enhanced immune-mediated clearance of transplanted human breast tumors when hematopoiesis was engineered for tumor-targeted IFN-α expression. By applying our strategy to mouse breast cancer models, we achieved inhibition of tumor progression and experimental metastases in an autologous setting, likely through enhanced generation of effector T cells and their recruitment to the neoplastic tissues. By forcing IFN-α expression in tumor-infiltrating macrophages, we blunted their innate protumoral activity and reprogrammed the tumor microenvironment toward more effective dendritic cell activation and immune effector cell cytotoxicity. Overall, our studies validate the feasibility, safety, and therapeutic potential of a new cancer gene therapy strategy, and open the way to test this approach as adjuvant therapy in advanced breast cancer patients