Kx 21n

At the end of treatment in the ethanol-sucrose-induced hypertensive rat model, blood samples from rats in the various treatment groups were obtained from the jugular vein using standard protocols [30 (link), 31 (link)] into labeled ethylenediaminetetraacetate (EDTA) tubes (Mediplus, Mumbai) for hematological profiling and gel separation tubes (Mediplus, Mumbai) for biochemical analysis. Full blood count was done at Noguchi Memorial Institute for Medical Research using an automated haematology analyser (SYSMEX KX-21N, USA) whilst liver and kidney function tests were performed on the samples at the ChemPath Laboratory, Mampong-Akuapem, using a fully automated chemistry analyser (PENTRA C200, Belgium).

Complete blood count (CBC) namely white blood cell (WBC), Diff count (neutrophil count (NEU#), lymphocyte count (LYM#), mixed cells count (MXD#), neutrophil percentages (NEU%), lymphocyte percentages (LYM%), mixed cells percentages (MXD%)) red blood cells (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red cell distribution width (RDW), platelets (PLT), mean platelet volume (MPV) and platelet distribution width (PDW) were analyzed using Sysmex KX-21 N (Sysmex Corporation, Kobe, Japan) [41 ] automated hematology analyzer. The analyzer performs 18 parameters within one minute. It utilizes three detector blocks for WBC (DC detection method), HGB (Non-cyanide HGB method), RBC and PLT (DC Detection method)). Peripheral blood smears from cord blood samples were prepared and stained using Wright’s stain for investigation of red blood cell morphology, white blood cell and platelets abnormalities.

To ensure leukemia development and engraftment, peripheral blood (PB) samples were taken from the tail of mice for complete blood counts using an automated cell counter (Sysmex KX-21N, Sysmex America, Inc., Mundelein, IL) and flow cytometry (FCM) (BD FACS Canto II, BD Biosciences, San Diego, CA). The spleens from dead or euthanized animals were analyzed for evidence of AML. Single-cell suspensions from PB samples were stained with fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD117, phycoerythrin (PE)-conjugated anti-mouse CD34 and allophycocyanin (APC)-conjugated anti-mouse Ly-6G and Ly-6C (myeloid differentiation antigen, Gr-1) (all from BD Biosciences Pharmingen, San Diego, CA). A minimum of 10,000 events were acquired for each sample by FCM and data analyzed using FACSDiva software (BD Biosciences, San Diego, CA).

Blood with anticoagulant was recollected for erythrocyte count (EC), hemoglobin (Hb) concentration, hematocrit (packed cell volume, PCV), and reticulocytes, using automated Coulter method with SYSMEX kx-21N® equipment.

This was a diagnostic accuracy study, separated into two phases, and distinguished by the location of the Presto device either in the district hospital DH (phase 1) or the rural lab (phase 2). In the first phase, all participants were recruited from the DH clinic, and samples were tested on-site. Results from the Presto were compared to the FACSCount (Becton Dickinson, CA, USA) for CD4 Count, and to the Sysmex KX– 21N (Sysmex Corporation, Japan) for hemoglobin, which were also operated at the DH. In the second phase, the Presto was operated in the rural laboratory, and results were compared again to the FACSCount and Sysmex, which were still operated at the DH. Participants in phase 2 were recruited from rural clinics, as well as from the DH. Samples from the DH were therefore transported to the rural laboratory for testing on the Presto. Using Presto at the rural laboratory provided insight into the feasibility of using near-patient CD4 counters in low-resource areas, as compared to the larger and better equipped laboratory at the DH. The Presto test results were never used for patient care in either phase, and all clinical decisions were made based on results from venous blood samples tested on the reference machines.