Phosphatidylserine and Tyro3-Axl-Mertk Receptor Tyrosine Kinase level detection in plasma and on plasma-derived extracellular vesicle surface in chronic lymphocytic leukemia.

Chronic lymphocytic leukemia (CLL) is a chronic lymphoproliferative disorder characterized by monoclonal B cell proliferation. Studies carried out in recent years suggest that extracellular vesicles (EVs) may be a potential biomarker in cancer. Tyro3-Axl-Mertk (TAM) Receptor Tyrosine Kinases (RTKs) and Phosphatidylserine (PS) have crucial roles in macrophage-mediated immune response under normal conditions. In the tumor microenvironment, these molecules contribute to immunosuppressive signals and prevent the formation of local and systemic antitumor immune responses. Based on this, we aimed to evaluate the amount of PS and TAM RTK in plasma and on the surface of EVs in CLL patients and healthy volunteers in this study. In this study, 25 CLL (11 F/14 M) patients in the Rai (O-I) stage, newly diagnosed or followed up without treatment, and 15 healthy volunteers (11 F/4 M) as a control group were included. For all samples, PS and TAM RTK levels were examined first in the plasma and then in the EVs obtained from the plasma. We detected a significant decrease in plasma PS, and TAM RTK levels in CLL patients compared to the control. Besides, we determined a significant increase in TAM RTK levels on the EV surface in CLL, except for PS. In conclusion, these receptor levels measured by ELISA in plasma may not be effective for the preliminary detection of CLL. However, especially TAM RTKs on the surface of EVs may be good biomarkers and potential targets for CLL therapies.

A CD34+ Cell Enrichment Protocol of Hematopoietic Stem Cells in a Well-Established Quality Management System.

Allogeneic stem cell transplantation applications have improved tremendously over the past quarter of a century. The use of new immunosuppressive protocols and elimination of T cells by CD34+ cell enrichment or T cell depletion on apheresis products increases the chance of using partially matched or haploidentical grafts. This is without increasing the risk of graft-versus-host disease, which is observed as a major complication of hematopoietic stem cell transplantation. The aim of this protocol is to evaluate the results obtained from 6 different process cycles performed on 6 different days. We used the CliniMACS Plus system located in our Cell and Tissue Manufacturing Center Quality Control Unit which is already calibrated as a class D room and includes a class A microbiological safety cabinet inside. The average purity of the end products was 95.66%, excluding only one end product which was 70%; this was higher than the values in current studies in the field. Superior to the reported studies, the CD3 quantity in each end product was below the dedicated thresholds. BactecTM FX40 blood culture system test results were detected as negative for each end product. Endotoxin testing suggested the absence of endotoxin within the products. The consistent outcomes obtained from these 6 different process cycles confirmed that the CliniMACS® Plus process cycles performed in accordance with our well-defined quality management system procedure is sufficient for the routine application of high-quality and safe CD34+ enrichment processes within our clean room area.

Comparison of the methods evaluating post thawing viability of peripheral blood stem cell graft.

OBJECTIVE: Reliable and pratique methods are essential for rapid and accurate determination of post thawing viability of peripheral blood stem cell (PBSC) graft before hematopoietic stem cell transplantation. In this study, Trypan Blue (TP) Eosin Y (EO), and Acridine-orange-ethidium bromide (AO/EB), which are of the methods commonly used for the assessment of viability in clinic practice, were compared with the flow cytometry-7AAD (7AAD) method, which is a more sensitive method. The aim of this study is to examine which method evaluates postthawing viability in a more compatible manner with 7AAD. MATERIALS-METHODS: Postthawing viability rates were examined simultaneously by means of four different methods before hematopoietic stem cell transplantation in a total of 20 PBSC graft. The results obtained from the AO/EB, TP, EO methods were evaluated with the flow cytometry-7AAD in terms of concordance. RESULTS: The AO / EB was determined to be the method having the best concordance with the flow cytometry-7AAD method. Although, at a lower level compared to the AO/EB method, the EO method had a statistically significant concordance with the flow cytometry-7AAD method. No statistically significant concordance was detected between the TP method and 7AAD method in terms of viability results. CONCLUSION: The AO/EB method was identified to be the method having the best compatibility with the flow cytometry -7AAD method in showing the viability of the cryopreserved PBSC graft. In the viability assessment of PBCS graft using light microscopy, the EO may be preferred since is more sensitive compared to the TP method.

Therapeutic plasmapheresis in geriatric patients: favorable results.

Therapeutic plasma exchange is generally a tolerable procedure, although several complications should be considered. Since geriatric population has been growing worldwide, the aim of this study was to retrospectively analyze 4709 TPE data from 981 geriatric procedures (20.8%) and to compare them with 3728 non-geriatric procedures (79.2%). The most common indications for TPE in both groups were sepsis/adult respiratory distress syndrome and multiple organ dysfunction. In geriatric patient group, contrary to expectations of aggravation, complication rate was statistically similar with non-geriatric group (P > 0.05). Therefore, TPE appeared to be a safe procedure in geriatric patients when performed by experienced practitioners.

rHuEPO hyporesponsiveness and related high dosages are associated with hyperviscosity in maintenance hemodialysis patients.

OBJECTIVE: Increased viscosity may increase the risk of thrombosis or thromboembolic events. Recombinant human erythropoietin (rHuEPO) is the key stone treatment in anemic ESRD patients with the thrombotic limiting side effect. We evaluated the influence of clinical and laboratory findings on plasma viscosity in MHD patients in the present study. METHOD: After applying exclusion criteria 84 eligible MHD patients were included (30 female, age: 54.7 ± 13.7 years). RESULTS: Patients with high viscosity had longer MHD history, calcium × phosphorus product, and higher rHuEPO requirement (356.4 versus 204.2 U/kg/week, P: 0.006). rHuEPO hyporesponsiveness was also more common in hyperviscosity group. According to HD duration, no rHuEPO group had the longest and the low rHuEPO dosage group had the shortest duration. Despite similar Hb levels, 68% of patients in high rHuEPO dosage group; and 38.7% of patients in low rHuEPO dosage group had higher plasma viscosity (P: 0.001). Patients with hyperviscosity had higher rHuEPO/Hb levels (P: 0.021). Binary logistic regression analyses revealed that rHuEPO hyporesponsiveness was the major determinant of hyperviscosity. CONCLUSION: We suggest that the hyperviscous state of the hemodialysis patients may arise from the inflammatory situation of long term HD, the calcium-phosphorus mineral abnormalities, rHuEPO hyporesponsiveness, and related high dosage requirements.

Therapeutic plasma exchange in treatment of neuroimmunologic disorders: review of 92 cases.

Therapeutic plasma exchange (TPE) is a procedure that reduces amount of circulating antibodies in patients through filtration for the treatment of neurologic diseases in which autoimmunity plays a major role. We reviewed the medical records of 92 neurologic patients who had been consecutively treated by TPE between June 2000 and April 2011 at Ankara University School of Medicine, Neurology Department and The Apaheresis Center. Neurological indications included myastehia gravis (MG, 16 patients), Guillain-Barre syndrome (GBS, 37 patients) and miscellaneous diseases (39 patients). The median TPE session number was 5 with a range of 1-8; total number of TPE procedures in all cases was 454. All MG patients improved with TPE during their hospitalization time. Regarding GBS, nearly 67% of the patients improved early, during their hospitalization time, either. In our series, 25% of GBS cases died because of dysautonomia. TPE was not effective in the treatment of the patients with Lambert-Eaton myasthenic syndrome, paraneoplastic polineuropathy, toxic polineuropathy, mononeuropathy multiplex in the case series. During the TPE procedures, 4 patients had hypotension and total number of the procedures was 21 in those patients. One patient had urticaria in only one session of total 5 TPE procedures. Two patients had septicemia; the first one had 3 and the second had 5 TPE procedures; both septic cases died. In conclusion, TPE is an effective treatment in neurologic diseases that autoimmunity plays an important role in the pathogenesis.

The effect of chronic cadmium toxicity on blood pressure and plasma viscosity.

Cadmium (Cd) is a heavy metal which affects many systems in humans and animals as a consequence of environmental and industrial pollution. The aim of this study was to investigate the effect of chronic Cd toxicity on blood pressure and plasma viscosity. Experimental group rats were given doses that contained 15 ppm CdCl(2) in drinking water for 8 weeks. The systolic blood pressure and heart rate were measured from rats' tails and recorded by plethysmography every two weeks. Blood samples were drawn, Cd levels were determined by atomic absorption spectrophotometer and plasma viscosity values by viscometer. Blood Cd levels were found to be significantly higher in the experimental group compared to the control group (p < 0.001). The whole blood analysis was made by an analyzer. Polymorphonuclear leukocytes and monocytes increased (p < 0.01) and lymphocyte number (p < 0.05) decreased in the experimental group. Viscosity values were 2.21 ± 0.54 and 1.62 ± 0.31 centipoises in the experimental and control groups, respectively (p < 0.001). In the experimental group, changes in systolic blood pressure between weeks were significant (p < 0.001) and were found to be correlated with plasma viscosity (p < 0.001). In the experimental group, changes in heart rate between weeks were significant (p < 0.001). According to our findings, Cd toxicity may lead to an increase in blood pressure by increasing plasma viscosity.