Bakry R.,Transfusion Center |
Sayed D.,South Egypt Cancer Institute |
Galal H.,Assiut University |
Shaker S.,South Valley University
Therapeutic Apheresis and Dialysis | Year: 2010
Platelets are known to undergo shape change, activation, release reaction and apoptosis/necrosis during processing and storage. Apheresis may have a deleterious impact on platelet achievability and functional integrity. Platelet concentrates from 50 male volunteers obtained by COBE spectra were screened for platelet activation (CD62 and CD154) and apoptosis (phosphatidylserine detected by Annexin V). Donor samples before separation, during apheresis and at the third day of storage were used as baseline donor samples. Platelet aggregation to adenosine diphosphate (ADP) and collagen was performed. There was a statistically significant increase in the expression of activation markers in two different samples (during separation samples and third day samples). Although the increase in Annexin V expression was not so observable, it showed a significant increase also. There was marked decline in the platelet aggregation. The correlations between the values of CD62, CD154 and Annexin V were detected in baseline samples and increased during separation and at the third day of platelets storage. Correlation between values of platelet aggregation to collagen and Annexin V was relevant only in the baseline samples. No other correlations were encountered between platelet aggregation and markers of activation and apoptosis during apheresis and storage. Initial platelet activation induced by apheresis may have an impact on phosphatidylserine expression with no impact on aggregation function of platelets during storage. © 2010 International Society for Apheresis.
Gringeri A.,University of Milan |
Lundin B.,Lund University |
Von Mackensen S.,University of Milan |
Von Mackensen S.,University of Hamburg |
And 18 more authors.
Journal of Thrombosis and Haemostasis | Year: 2011
Background: Prevention of arthropathy is a major goal of hemophilia treatment. While studies in adults have demonstrated an impact of prophylaxis on the incidence of joint bleeds and patients' well-being in terms of improved quality of life (QoL), it is unclear whether or not prophylaxis influences the outcome and perception of well- of children with hemophilia. Objective: This randomized controlled study compared the efficacy of prophylaxis with episodic therapy in preventing hemarthroses and image-proven joint damage in children with severe hemophilia A (factor VIII < 1%) over a 10-year time period. Methods: Forty-five children with severe hemophilia A, aged 1-7 years (median 4), with negative clinical-radiologic joint score at entry and at least one bleed during the previous 6 months, were consecutively randomized to prophylaxis with recombinant factorVIII (25 IU kg-1 3 × week) or episodic therapy with ≥ 25 IU kg-1 every 12-24 h until complete clinical bleeding resolution. Safety, feasibility, direct costs and QoL were also evaluated. Results: Twenty-one childrenwere assigned to prophylaxis, 19 to episodic treatment. Children on prophylaxis had fewer hemarthroses than children on episodic therapy: 0.20 vs. 0.52 events per patient per month (P < 0.02). Plainfilm radiology showed signs of arthropathy in six patients on prophylaxis (29%) vs. 14 on episodic treatment (74%) (P < 0.05). Prophylaxis was more effective when started early (≤ 36 months), with patients having fewer joint bleeds (0.12 joint bleeds per patient per month) and no radiologic signs of arthropathy. Conclusion: This randomized trial confirms the efficacy of prophylaxis in preventing bleeds and arthropathy in children with hemophilia, particularly when it is initiated early in life. © 2011 International Society on Thrombosis and Haemostasis.
Pizon M.,Transfusion Center |
Zimon D.,Transfusion Center |
Carl S.,Friedrich - Schiller University of Jena |
Pachmann U.,Transfusion Center |
And 2 more authors.
ecancermedicalscience | Year: 2013
Background: The detection of tumour cells circulating in the peripheral blood of patients with breast cancer is a sign that cells have been able to leave the primary tumour and survive in the circulation. However, in order to form metastases, they require additional properties such as the ability to adhere, self-renew, and grow. Here we present data that a variable fraction among the circulating tumour cells detected by the Maintrac® approach expresses mRNA of the stem cell gene NANOG and of the adhesion molecule vimentin and is capable of forming tumour spheres, a property ascribed to tumour-initiating cells (TICs). Patients and methods: Between ten and 50 circulating epithelial antigen-positive cells detected by the Maintrac approach were selected randomly from each of 20 patients with breast cancer before and after surgery and were isolated using automated capillary aspiration and deposited individually onto slides for expression profiling. In addition, the circulating tumour cells were cultured without isolation among the white blood cells from 39 patients with breast cancer in different stages of disease using culture methods favouring growth of epithelial cells. Results: Although no epithelial cell adhesion molecule (EpCAM)-positive cells expressing stem cell genes or the adhesion molecule vimentin was detected before surgery, 10%-20% of the cells were found to be positive for mRNA of these genes after surgery. Tumour spheres from circulating cells of 39 patients with different stages of breast cancer were grown without previous isolation in a fraction increasing with the aggressivity of the tumour. Summary: Here we show that among the peripherally circulating tumour cells, a variable fraction is able to express stem cell and adhesion properties and can be grown into tumour spheres, a property ascribed to cells capable of initiating tumours and metastases. Copyright: © the authors.
PubMed | Director Transfusion Center and Transfusion Center
Type: Journal Article | Journal: Immunohematology | Year: 2015
Classic methods to determine human red blood cell (RBC) antigens are based on serologic testing. Thanks to increased knowledge of the molecular basis associated with many blood group antigens, it is currently possible to predict their presence or absence on the red cell membrane. Several molecular techniques have been developed to detect the most important allelic variations attributable to single nucleotide polymorphisms. The human erythrocyte antigen (HEA) BeadChip system manufactured by BioArray Solutions (Immucor, Warren, NJ) is one of the commercial DNA array platforms currently available to predict HEAs by DNA analysis. This technology provides a useful tool to increase the inventory of antigen-negative RBC units and prevent immunization of patients who require chronic transfusion by providing compatible RBC units based on matching by DNA testing.
Martelli A.M.,University of Bologna |
Martelli A.M.,CNR Institute of Molecular Genetics |
Papa V.,University of Cassino and Southern Lazio |
Tazzari P.L.,Transfusion Center |
And 13 more authors.
Leukemia | Year: 2010
Alkylphospholipids and alkylphosphocholines (APCs) are promising antitumor agents, which target the plasma membrane and affect multiple signal transduction networks. We investigated the therapeutic potential of erucylphosphohomocholine (ErPC3), the first intravenously applicable APC, in human acute myelogenous leukemia (AML) cells. ErPC3 was tested on AML cell lines, as well as AML primary cells. At short (6-12 h) incubation times, the drug blocked cells in G2/M phase of the cell cycle, whereas, at longer incubation times, it decreased survival and induced cell death by apoptosis. ErPC3 caused JNK 1/2 activation as well as ERK 1/2 dephosphorylation. Pharmacological inhibition of caspase-3 or a JNK 1/2 inhibitor peptide markedly reduced ErPC3 cytotoxicity. Protein phosphatase 2A downregulation by siRNA opposed ERK 1/2 dephosphorylation and blunted the cytotoxic effect of ErPC3. ErPC3 was cytotoxic to AML primary cells and reduced the clonogenic activity of CD34+ leukemic cells. ErPC3 induced a significant apoptosis in the compartment (CD34+ CD38 Low/Neg CD123+) enriched in putative leukemia-initiating cells. This conclusion was supported by ErPC3 cytotoxicity on AML blasts showing high aldehyde dehydrogenase activity and on the side population of AML cell lines and blasts. These findings indicate that ErPC3 might be a promising therapeutic agent for the treatment of AML patients. © 2010 Macmillan Publishers Limited All rights reserved.
Pachmann K.,Transfusion Center
Expert Review of Molecular Diagnostics | Year: 2015
Cells shed from solid malignant tumors into the circulation are considered to be the origin of metastases. In spite of a wealth of research on the pathway of metastasis formation, it is still not clear when and how metastases develop, nor is there a consensus on the number and the nature of circulating tumor cells present in individual patients and their relationship to the formation of metastases. We have developed a method to detect a maximum of unselected non-hematological, epithelial cells in the blood, assuming that in cancer patients the majority of these cells are derived from the tumor. Assessment of the number of these cells longitudinally during the course of disease and therapy allows the response to different treatments to be monitored. Due to the viability of the cells, additional analyses such as expression profiles and determination of their sensitivity to drugs can be performed. © Informa UK, Ltd.
Angelini M.,Clinic of Obstetrics and Gynecology |
Barillari G.,Transfusion Center |
Londero A.P.,Clinic of Obstetrics and Gynecology |
Bertozzi S.,AOU Santa Maria della Misericordia |
And 4 more authors.
Journal of Thrombosis and Thrombolysis | Year: 2013
Ovarian vein thrombosis (OVT) is an uncommon but potentially serious complication in the early postpartum. Two case studies seem to prove the point: Case 1 A 24-year-old woman was transferred to our hospital with the chief complaint of abdominal pain radiating to the right thigh, vomit, diarrhea, and a slight pyrexia (37.6 C rectal). Five days earlier, she had a spontaneous vaginal delivery after labor induction. The woman appeared slightly distressed because of pain; vital signs were found to be normal and the CRP elevated (129.9 mg/L). Abdominal examination was remarkable for tenderness by palpation in the right lower quadrant with no rebound tenderness or guarding. Pelvic examination was remarkable for mild right adnexal tenderness. Abdominal-pelvic computer tomography with contrast medium revealed a 2.5-cm OVT having extended into the inferior vena cava for 14 cm with a slight peripheral edema. The patient was treated with nadroparin 0.6 cc (5700 IU) bid and warfarin 5 mg since the attainment of the therapeutic INR range. Case 2 A 31-year-old twin-pregnant woman had an emergency cesarean section at 35 gestational weeks because of hypertension complicated by increased liver enzymes, diuresis contraction, and continuous lower back pain bilaterally radiating to the groins. One day after delivery, CT scan that was performed because of onward anemia showed a pelvic, perihepatic, and perisplenic blood effusion, and a 1-cm right OVT extended to the inferior vena cava below renal veins for 28 mm. She underwent exploratory laparotomy and blood transfusion, and because of respiratory insufficiency she was transferred to a second level center with ICU facility, where she was placed under a suprarenal inferior vena cava filter, and AngioJet Rheolytic Thrombectomy for acute pulmonary embolism was performed. © 2012 Springer Science+Business Media, LLC.
Vagace J.M.,Materno Infantil Hospital |
Rodriguez M.A.,Transfusion Center |
De La Maya M.D.,Materno Infantil Hospital |
Gervasini G.,University of Extremadura
Journal of Clinical Pathology | Year: 2013
We investigated the case of a 14-year-old girl with an ethylenediaminetetraacetic acid (EDTA)-dependent haemagglutination detected by macrocytosis, which was only evident by an abnormal red blood cell (RBC) population in the histogram. Investigations included haemograms with different anticoagulants and experimental conditions. Immunohaematological studies were performed using a gel-based technology. At admission, the patient had a low RBC count and an increased mean corpuscular volume with normal haemoglobin. A double population appeared in the RBC histogram. However, the peripheral blood smear was normal and macrocytosis was absent when heparin or citrate was used instead of EDTA. Later studies revealed that the patient's serum was able to induce macrocytosis of control RBC only in the presence of EDTA. An EDTAdependent panagglutinin was then indentified that produced mixed field agglutination. These findings provide evidence of a haemagglutination induced by EDTA as a source of pseudomacrocytosis.
PubMed | Ontario Cancer Institute and Transfusion Center
Type: Journal Article | Journal: Hematology (Amsterdam, Netherlands) | Year: 2016
Adhesion of hematopoietic precursors to the marrow microenvironment appears to be a prerequisite for proliferation and differentiation of hematopoietic cells. In this report, we have studied the adhesion of CFU-GM from marrow CD34+ precursors to human marrow myofibroblasts and to an human stromal cell line, L2Ori-, transformed by a vector comprising the whole of the SV40 viral sequence except for the origin of replication. This Stro-l(+) cell line presents characteristics similar to those of vascular smooth muscle cells, since (i) few cells were -SM actin(+)while all cells were vimentin(+) but desmine(-) and a metavinculin band was consistently detected, (ii) all cells contained lysosomes filled with glycoproteins recognized by the monoclonal antibody 1B10, (iii) we detected EDa(+) EDb(-) pericellular fibronectin and intracellular 1 and laminins and (iv) the cytokine expression pattern was similar to that of cells from colony-derived cell lines. Transformation was confirmed by abnormal and irregular growth (hallmarked by crises with rather slow growth between crises), and the presence of some very large cells with several nuclei. Although presenting an usual stromal phenotype, this cell line could not sustain hematopoiesis from marrow CD34+ cells in coculture due to a complete inability of adhesion of CD34+ cells (0% of adherent CFU-GM vs. 20% on normal stromal myofibroblasts). The lack of adhesion was explained by abnormal expression of adhesion molecules and molecules involved in the organization of extracellular matrix: (1) at the membrane level: the lack of VCAM-1 and significant differences in the distribution of CD44 and integrins 1, 3, 4 and as compared to normal stroma; (2) at the level of focal adhesions: the predominance of the 200 kD fragment of talin (as opposed to that of 230 kD in normal stroma), and a significantly decreased expression of vinculin and -actinin; (3) at the level of microfilaments: the decrease in polymerized actin and a large decrease of -SM actin synthesis; and (4) at the level of extracellular matrix: very few fibronectin fibres. These data show that transformation can strongly and negatively affect the function of hematopoiesis maintenance by disrupting intercellular and extracellular matrix adhesion mechanisms of hematopoietic cells to the stroma, in particular by affecting the fibronexus. Our data suggest the need for extreme caution when using SV40 transformed cell lines and instead, make the case for the use of other means of immortalization (such as thermosensitive T, other transforming sequences, introduction of inducible promoters).
PubMed | Transfusion Center
Type: Journal Article | Journal: Expert review of molecular diagnostics | Year: 2015
Cells shed from solid malignant tumors into the circulation are considered to be the origin of metastases. In spite of a wealth of research on the pathway of metastasis formation, it is still not clear when and how metastases develop, nor is there a consensus on the number and the nature of circulating tumor cells present in individual patients and their relationship to the formation of metastases. We have developed a method to detect a maximum of unselected non-hematological, epithelial cells in the blood, assuming that in cancer patients the majority of these cells are derived from the tumor. Assessment of the number of these cells longitudinally during the course of disease and therapy allows the response to different treatments to be monitored. Due to the viability of the cells, additional analyses such as expression profiles and determination of their sensitivity to drugs can be performed.