Cancer diagnoses profile of bone marrow specimens from November 2012 to December 2015 in a tertiary academic setting

This study aimed to examine the number and type of cancer diagnoses made based on bone marrow aspirates (BMA) using morphology and/or histology, over a three year period. All the data collected (2012-2015) was obtained using the Trak-Care laboratory information system (LIS) of the National Health Laboratory Service (NHLS). Data was categorised into four categories including acute leukaemias (ALs), chronic leukaemias (CLs), Hodgkin’s lymphoma (HL), Non-Hodgkin’s lymphoma (NHL) and a miscellaneous group. The laboratory test most frequently used to make diagnoses was bone marrow aspiration (BMA) morphology (199), followed by flow cytometry (138), histology (113), fluorescent in situ hybridisation (FISH) (54) and polymerase chain reaction (PCR) (9). In total, the top three conditions diagnosed were acute myeloid leukaemias (AMLs), chronic myeloid leukaemia (CML) and B-cell acute lymphoblastic leukaemia (B-ALL). There was good agreement between the diagnoses made by BMA morphology, BMT histology and flow cytometry. Results showed that BMA morphology was the most popular diagnostic test used and that this test had excellent agreement with BMT histology and flow cytometry diagnoses. The most frequently diagnosed conditions were the AMLs.http://www.smltsa.org.za/publications.htmlam2018Haematolog

Retrospective data analysis of all requests for flow cytometric immunophenotyping in a tertiary hospital setting

Flow cytometry is a globally accepted diagnostic tool used for the rapid identification of cells based on their surface and intracellular antigens, especially for the diagnosis of haematological malignancies. The aim of this study was to evaluate the requests received for flow cytometric immunophenotyping and to create a profile of diagnoses. In 2014 data regarding indications and diagnoses were captured from request forms received and final diagnosis reports issued by the Tshwane Academic Division (TAD) of the National Health Laboratory Service (NHLS). A total of 1234 requests were received over the one year period, of which 80.4% were performed and 16.8 % were rejected. The most common indications were leukaemia, lymphoma and cytopenia. Nineteen percent of requests received contained no correct indication or clinical history. In total, 103 and 153 diagnoses were established based on peripheral blood and bone marrow aspirate specimens respectively. Samples were mostly rejected due to sample clotting, electronic gate keeping rules and receiving the specimen more than 24 hours after collection.http://www.smltsa.org.za/publications.htmlam2018Haematolog

The impact of collection tube fill volume on international normalized ratio

INTRODUCTION: Pre-analytical variability currently represents the most important source of errors that can lead to inaccurate patient results in monitoring of patients being treated with oral anticoagulant therapy. The volume of blood collected is critical for accurate coagulation results. The National Committee for Clinical Laboratory Standards (NCCLS) recommends a ratio of blood to anticoagulant volume of 9:1. However, investigators have published reports which suggest that a lower ratio may be acceptable. Unfortunately the recommendations of these reports are inconsistent. AIM: The aim of this study was to determine the impact of tube fill volume on INR values both in healthy subjects and patients receiving oral anticoagulation therapy. METHODS: INR values were obtained by processing coagulation specimens containing different volumes of whole blood. The study group included 30 patients taking oral anticoagulation therapy and 15 healthy volunteers. Respectively 2.5ml, 3 ml, 3.5 ml, 4 ml and 4.5 ml of whole blood was drawn into tubes containing a fixed volume of 3.2% (0.109M) sodium citrate. RESULTS: The INR values increased as total tube fill volumes decreased for both groups but this finding did not reach statistical significance in either group for the tube fill volumes studied. CONCLUSION: For blood specimens collected in 3.2% citrate anticoagulant, a total tube fill volume of greater than 56% yielded reliable INR resultshttp://www.smltsa.org.z

Apoptotic profiling of chronic myeloid leukaemia patients' platelets ex vivo before and after treatment with Imatinib

Chronic myeloid leukaemia (CML) is a malignancy of the haematopoietic stem cells. The first line of treatment for CML, especially in developing countries, remains the first-generation tyrosine kinase inhibitor, Imatinib. Patients with CML are frequently diagnosed with platelet abnormalities. However, the specific mechanism of platelet abnormalities in CML remains unclear and poorly understood. The aim of this study was therefore to determine the apoptotic profiles of CML patients ex vivo on platelets before and after treatment with Imatinib. Blood samples of healthy volunteers and CML patients at diagnosis and after 6 months treatment with Imatinib were collected. Platelet counts, viability and activation were determined. Results showed that CML patients' platelet counts were elevated upon diagnosis and these levels statistically significantly decreased after 6 months of treatment. Platelet activation was significantly increased after 6 months of treatment compared to levels at diagnosis (P-value < .05). Similarly, platelet apoptosis was also increased after 6 months of treatment. Abnormalities in platelet functioning found in this study may partly be due to clonal proliferation of haematopoietic cells in CML patients, specifically of megakaryocyte precursors as well as the inhibition of platelet tyrosine kinase's and the inhibition of platelet-derived growth factor.Cancer Association of South Africa; Medical Research Council of South Africa; National Research Foundation; School of Medicine Research Committee of the Faculty of Health Sciences, University of Pretoria; Struwig-Germeshuysen Research Trust.http://wileyonlinelibrary.com/journal/cbfhj2022HaematologyInternal MedicinePhysiolog

Neutrophil extracellular traps and their role in health and disease

The human innate immune system is indispensable for protection against potentially invasive microbial and viral pathogens, either neutralising them or containing their spread until effective mobilisation of the slower, adaptive (specific), immune response. Until fairly recently, it was believed that the human innate immune system possessed minimal discriminatory activity in the setting of a rather limited range of microbicidal or virucidal mechanisms. However, recent discoveries have revealed that the innate immune system possesses an array of novel pathogen recognition mechanisms, as well as a resourceful and effective alternative mechanism of phagocyte (predominantly neutrophil)-mediated, anti-infective activity known as NETosis. The process of NETosis involves an unusual type of programmed, purposeful cell death, resulting in the extracellular release of a web of chromatin heavily impregnated with antimicrobial proteins. These structures, known as neutrophil extracellular traps (NETs), immobilise and contribute to the eradication of microbial pathogens, ensuring that the anti-infective potential of neutrophils is sustained beyond the lifespan of these cells. The current review is focused on the mechanisms of NETosis and the role of this process in host defence. Other topics reviewed include the potential threats to human health posed by poorly controlled, excessive formation of NETs, specifically in relation to development of autoimmune and cardiovascular diseases, as well as exacerbation of acute and chronic inflammatory disorders of the airways.http://www.sajs.co.zaam2016HaematologyImmunologyInternal Medicin

Pneumolysin mediates heterotypic aggregation of neutrophils and platelets <i>in vitro</i>

OBJECTIVES: Platelets orchestrate the inflammatory activities of neutrophils, possibly contributing to pulmonary and myocardial damage during severe pneumococcal infection. This study tested the hypothesis that the pneumococcal toxin, pneumolysin (Ply), activates production of platelet-activating factor (PAF) and thromboxane A2 (TxA2) by neutrophils, these bioactive lipids being potential mediators of neutrophil:platelet (NP) networking. METHODS: The effects of recombinant Ply (10–80 ng mL−1) on the production of PAF and TxA2 by isolated neutrophils were measured using ELISA procedures, and NP aggregation by flow cytometry. RESULTS: Exposure of neutrophils to Ply induced production of PAF and, to a lesser extent, TxA2, achieving statistical significance at ≥20 ng mL−1 of the toxin. In the case of NP interactions, Ply promoted heterotypic aggregation which was dependent on upregulation of P-selectin (CD62P) and activation of protease-activated receptor 1 (PAR1), attaining statistical significance at ≥10 ng mL−1 of the toxin, but did not involve either PAF or TxA2. CONCLUSION: Ply induces synthesis of PAF and TxA2, by human neutrophils, neither of which appears to contribute to the formation of NP heterotypic aggregates in vitro, a process which is seemingly dependent on CD62P and PAR1. These pro-inflammatory activities of Ply may contribute to the pathogenesis of pulmonary and myocardial injury during severe pneumococcal infection.South African National Research Foundation (NRF)http://www.elsevierhealth.com/journals/jinf2018-06-30hj2017HaematologyImmunologyInternal Medicin

An endometrial histomorphometric study of CD56+ natural killer cells in women with unexplained infertility

BACKGROUND. The number of peripheral blood and endometrial natural killer cells varies greatly during implantation and the first trimester of pregnancy and is thought to play a role in the maintenance of a healthy pregnancy. However, the role of endometrial CD56+ natural killer (NK) cells as an immunological mechanism in unexplained infertility is yet unknown. OBJECTIVES. The study aimed to enumerate the concentrations of CD56+ NK cells in endometrial samples, and to statistically compare these numbers between fertile and infertile women. METHODS. A histomorphometric analysis was conducted using haematoxylin and eosin staining and an immunohistochemical approach was used for quantifying cell numbers. RESULTS. Fifty samples were collected in equal parts between a study group of infertile female subjects (mean (standard deviation) age 35 (4), range 26 - 42 years) and a control group of multiparous fertile individuals (mean (SD) age 43.4 (6.3), range 30 - 55). The mean number of CD56+ NK cells present at different depths for both the study and control groups did not differ significantly. Age and group (study or control) were not significantly related to the mean number of CD56+ NK cells. However, for the late secretory phase the mean number of CD56+ NK cells was significantly higher than for the early phase. CONCLUSION. Our findings could not identify a statistically significant correlation between the number of CD56+ NK cells and infertility.Discovery Foundation Research Awardshttp://sajog.org.za/index.php/SAJOGhttp://www.journals.co.za/content/journalam2017Anatomical PathologyAnatomyHaematologyImmunologyStatistic

Pan-Cancer Analysis of lncRNA Regulation Supports Their Targeting of Cancer Genes in Each Tumor Context

Long noncoding RNAs (lncRNAs) are commonly dys-regulated in tumors, but only a handful are known toplay pathophysiological roles in cancer. We inferredlncRNAs that dysregulate cancer pathways, onco-genes, and tumor suppressors (cancer genes) bymodeling their effects on the activity of transcriptionfactors, RNA-binding proteins, and microRNAs in5,185 TCGA tumors and 1,019 ENCODE assays.Our predictions included hundreds of candidateonco- and tumor-suppressor lncRNAs (cancerlncRNAs) whose somatic alterations account for thedysregulation of dozens of cancer genes and path-ways in each of 14 tumor contexts. To demonstrateproof of concept, we showed that perturbations tar-geting OIP5-AS1 (an inferred tumor suppressor) andTUG1 and WT1-AS (inferred onco-lncRNAs) dysre-gulated cancer genes and altered proliferation ofbreast and gynecologic cancer cells. Our analysis in-dicates that, although most lncRNAs are dysregu-lated in a tumor-specific manner, some, includingOIP5-AS1, TUG1, NEAT1, MEG3, and TSIX, synergis-tically dysregulate cancer pathways in multiple tumorcontexts