Direct-Push Blood Transfusion Practice in the Neonatal Unit of a Tertiary Hospital in South-South, Nigeria

Introduction: Transfusion of blood is a life-saving intervention in the care of ill neonates. Donated blood is a scarce national resource and must be used in the most efficient way. Exchange blood transfusion using the blood bag is the commonest mode of blood delivery employed. Other modalities of safe and sustainable blood delivery should also be explored, especially where paucity of funds predominates. This study aims to assess the usefulness of the direct push method where applicable, as an alternative to blood bag delivery in neonatal units of resource poor settings. Methods: A two year retrospective study of newborns admitted in the neonatal wards of the University of Uyo Teaching Hospital. Data obtained were the age, gender, indication for admission, packed cell volume (PCV) before and after transfusion. Blood transfusion was done in aliquots over 24 hours under aseptic conditions, via a peripheral vein. The push and pull method was employed, with no anticoagulant in the syringe. Post-transfusion PCV was done at least 24 hours after the procedure. Results: Of the one thousand and seventy-seven (1077) admitted neonates, two hundred and thirty-nine (22.2%), received blood products. Of these, twenty-one (8.8%), received a direct whole blood transfusion. Age (days) of the neonates transfused ranged from 1 to 26 days, with a mean of 10.4 ± 8.13. The Packed Cell Volume (PCV) pre-transfusion ranged between 20% - 44%, with a mean of 30.05 ± 6.39 while post-transfusion PCV ranged between 31% to 51%, with a mean of 38.17 ± 5.52(Fig. 1). The commonest indication for transfusion was prematurity, 9(42.8%) and neonatal sepsis 5 (23.8%). Conclusion: The direct transfusion of blood occasionally used, seems a relatively safe practice to correct mild/moderate anaemia. It also provides sufficient blood, with the advantage of usage when the umbilical cord access is no longer feasible and where cost of blood would otherwise, hinder quick intervention. This practice may need further evaluation by other centers.

Description of The Chemiluminescence Immunoassay (ChLIA) Method of HIV Screening Test on The Donor's Blood

Human Immunodeficiency Virus (HIV) is one of the transmitted infection through blood transfusion. Chemiluminescence immunoassay for HIV testing is performed to ensure the safety and quality of blood product that released. Purpose of the study to identify the result of Chemiluminescence Immunoassay methode for HIV testing to blood donor’s characteristic based on gender, age, blood group, HIV titer, and donation location. The design of this research is descriptive retrospective. There is 11 (0,8%) reactive blood bag and 1.289 (99,2%) blood bag non reactive to HIV, reactive blood bag based of gender is 8 (72,7%) blood bag from male donors and 3 (27,3%) blood bag from female donors. Based on donors age, mostly reactive result are from 41-70 years old is 4 (36,4%) donors, and the fewest is from (under) 19 years old is 1 (9,1%) donors. The majority based on blood group is B Rh+ is 4 (36,4%) donors. The majority based on HIV titer is low titer, 1,0-2,0 is 7 (63,6%) donors. Based on donation location, mostly from mobile unit (MU) is 8 (72,7%) donors over 11 donors with HIV reactive. Conclusion of this study is donors gender, donors age, and donation location had a significant influence on reactive result of HIV testing.

Blood Unit Segments Accurately Represent the Biophysical Properties of Red Blood Cells in Blood Bags but Not Hemolysis

BACKGROUND: The biophysical properties of red blood cells (RBCs) provide potential biomarkers for the quality of donated blood. Blood unit segments provide a simple and non-destructive way to sample RBCs in clinical studies of transfusion efficacy, but it is not known whether RBCs sampled from segments accurately represent the biophysical properties of RBCs in blood bags. STUDY DESIGN AND METHODS: RBCs were sampled from blood bags and segments every two weeks during 8 weeks of storage at 4 degrees C. RBC deformability was measured by deformability-based sorting using the microfluidic ratchet device in order to derive a rigidity score. Standard hematological parameters, including mean corpuscular volume (MCV), red cell distribution width (RDW), mean cell hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and hemolysis were measured at the same time points. RESULTS: Deformability of RBCs stored in blood bags was retained over 4 weeks storage but a progressive loss of deformability was observed at weeks 6 and 8. This trend was mirrored in blood unit segments with a strong correlation to the blood bag data. Strong correlations were also observed between blood bag and segment for MCV, MCHC and MCH, but not for hemolysis. CONCLUSION: RBCs sampled from blood unit segments accurately represents the biophysical properties of RBCs in blood bags, but not hemolysis. Blood unit segments provide a simple and non-destructive sample for measuring RBC biophysical properties in clinical studies.

Malaria Seroprevalence in the Blood Donors of Malaria Endemic Regions of Northern Andhra Pradesh, India.

Introduction: Transmission of malaria through blood transfusion continues to be a major threat tosafe blood transfusion practice. Transfusion-transmitted malaria occurs at an estimated rate of 0.25cases per 1 million blood units collected. It is significantly more common in endemic areas. Aim: Tostudy the Seroprevalence of Malaria among the blood donors in the endemic areas of NorthernAndhra Pradesh. Materials and methods: The present survey was carried out at the blood bank ofMaharaja institute of medical sciences, Vizianagaram. This includes the analysis of seroprevalence ofMalaria in the blood donors during the period of 1 year from February 2018 to January 2019. Two mlof the blood sample was collected in the labeled pilot tube at the time of collection of blood fromdonor tubing of the blood bag. The serum was separated. The samples were tested for Malaria byrapid antigen detection test. Results: Out of the total of 3096 blood donors, replacement donors(86.91%) were more in comparison to voluntary donors (13.08%). The seroprevalence among thereplacement blood donors was more compared to voluntary blood donors. Conclusion: Voluntaryblood donation, increasing awareness about blood donation in the general population, selection ofrepeat, non-remunerated, regular voluntary blood donors and diligent donor selection, sensitivescreening tests are most important to increase blood safety and prevent transmission of Malariathrough blood transfusion.

To Determine the Morphological Changes in Red Blood Cells During Storage in Blood Banks

Background: During storage of blood, the red blood cells undergo shape changes which cause fragility and endothelial interaction leading to deterioration the quality of blood in blood banks.Objectives: The aim of this study is to determine the morphological changes in red blood cells during storage in blood banks. Material and Methods: In this experimental study, a total 20 healthy volunteers between 17 to 40 years blood donors-Blood bags were taken, ten from each center i.e. MMCTH blood bank Mardan and KTH blood bank Peshawar. The specimen analysis was done at IBMS (Institute of Basic Medical Sciences) of KMU (Khyber Medical University) Peshawar. The exclusion criteria were People with anemia, hepatitis B &C, HIV and syphilis. The duration of this study was six months. The inform consent was taken from each donor. The total blood 250 ml from vein in cubital fossa from each blood donor was collected in 250ml pediatric blood bag with CPDA-1 solution. Blood bags were put up in the blood bank at +2 to +6 °C and stored till 20 days. Blood specimen of about 5cc were collected in 5cc syringe from each blood bag on 0, 5th,10th ,15th and 20th day for following parameters and thin film red blood cell was prepared for examination by light microscope. Morphological changes in RBCs examined via light microscope as well as grading the RBCs status in the peripheral blood film, the occurrence of distorted RBC simply in random fields; such as +1(scored 1 to 5 altered RBC present in each field), +2 (an average of 6 to 15 altered RBC in each field), +3(16 to 25 altered RBC in each field) and +4(more than 25 altered RBC present in each field). The multi head light microscope NIKON eclipse 50 was used for examination of peripheral blood slide and we took images of randomly selected field. The image J software was used for slide examination.Results: The morphological analysis of red blood cells, count of 200 cells in each blood slide in randomly selected fields are: On day 0 the majority of cells were normally shaped (97.95±1.297 (mean±SD).With increasing storage time, the percentage of morphologically abnormal red cells rose sharply. Mean percentage of abnormal cells on day 5, 10, 15 and 20 was 28.80±10.00, 51.73±12.47, 64.78±14.66 and 68.10±7.92 respectively. This increase in percentage of abnormally shaped cells was significant as determined by one way ANOVA (p =0.001). There was a big difference of percentage of abnormal RBCs on day 0 and in = 5 to= 10 days and in = 15 to = 20 days of blood storage. The mean values of day 0 of abnormal cells was 2.05±1.297 (Mean ± Std. Deviation), abnormal cells in= 5 to= 10 days was 40.26± 16.101 (Mean ± Std. Deviation) and on day = 15 and in = 20 day was 66.44± 11.75. The mean difference from day 0 to day 20 was 63.93±10.45 (Mean ± Std. Deviation).The one way ANOVA was significant, P= 0.001.Conclusion: This study confirms the hematological and morphological changes, when blood stored at 2 °C to 6 °C for up to 21 days. The significant morphological changes were observed on 5th day of blood storage. These findings suggested that approximately a week old stored blood is as good as the fresh blood; however, significant morphological and biochemical changes begin to appear after the first week of storage and these changes aggravate with time. Hence in order to achieve best possible transfusion outcomes, stored blood up to one week can be utilized.

The Effects of Pre-Storage Leukoreduction on the Conservation of Bovine Whole Blood in Plastic Bags

Leukoreduction (LR) is a technique that consists of reducing the number of leukocytes in whole blood or blood components that can contribute to decreasing storage lesions and the occurrence of post-transfusion complications. We propose that using a blood bag with pre-storage leukocyte filtration is sufficient for blood conservation under field conditions. Ten healthy Nelore cows were used. Whole blood was sampled from each animal and stored at 2 to 6 °C in CPD/SAG-M (citrate phosphate dextrose bag with a saline, adenine, glucose, mannitol satellite bag) triple bags (Control) and in CPD/SAG-M quadruple bags with a leukocyte filter (Filter). At baseline and after 7, 14, 21, 28, 35, and 42 days (D0, D7, D14, D21, D28, D35, and D42, respectively), complete hematological, blood gas, and biochemical evaluations were determined. The filtered bag removed 99.3% of white blood cells from cattle blood, and the entire filtration process was performed in the field. There was a reduction in the number of red blood cells (RBCs) in both groups from D14 onward, with a decrease of 19.7% and 17.1% at D42 for the Control and Filter bags, respectively. The hemoglobin (Hb) concentration had variation in both groups. Potassium, pO2, pCO2, and sO2 increased, and sodium, bicarbonate, and pH decreased during storage. The filtered bag was efficient in removing white cells from cattle whole blood and could be used under field conditions. Blood stored after LR showed differences (p < 0.05) in blood gas analysis towards a better quality of stored blood (e.g., higher pH, lower pCO2, higher sO2). Further experimental studies are required to prove that blood without white cells results in a decrease in transfusion reactions in cattle.

A Simple, Safe and Effective Approach for Personalized Precision Ex Vivo Gene Therapy Based on Autoinfusion of Gene-Modified Leucoconcentrate (GML) Prepared from Routine Unit of Patient's Peripheral Blood

Nowadays gene and cell therapy become the basic methods in regenerative medicine. However only few gene and cell products are currently approved for clinical usage. Biosafety problems, complexity of cell and gene technologies and high cost of manufacturing are the main reasons for the slow introduction of such approaches in practical medicine. Treatment of hereditary diseases of the immune system based on the correction of the mutant gene by delivering functional recombinant gene into WBC is the first successfully employed in the clinical practice approach of cell-mediated or ex vivo gene therapy. Earlier we have reported the strategy of the cell-mediated gene therapy based on umbilical cord blood mononuclear cells transduced with adenoviral vectors carrying recombinant genes encoding neurotrophic factors for treatment neurodegenerative diseases, neurotrauma and stroke. Significant disadvantage of this method is the usage of the umbilical cord blood mononuclear cells as a cell carrier for the therapeutic genes. Considering immunodeficiency treatment and our own data we developed a new approach of recombinant gene delivery for personalized ex vivo gene therapy. The method is based on autoinfusion of patient's WBC transduced with recombinant therapeutic genes for correction of certain pathological conditions. In the present study for the first time the human gene-modified leucoconcentrate (GML) producing recombinant reporter gene encoding green fluorescent protein (GFP) was obtained without culturing WBC in vitro. The routine unit of peripheral blood (450 ml) was collected into the plastic blood bag and the leucocyte- and platelet-rich concentrates (50 ml) were obtained by standard method using Macopress Smart (Macopharma, France). Afterwards the equal volume of hydroxyethyl starch 6% was added into the plastic blood bag which was centrifuged (DP-2065 R PLUS, Centrifugal Presvac RV; Presvac, Buenos Aires, Argentina) at 350 rpm for 10 min at 10°C. The obtained supernatant was transferred into the new plastic blood bag using manual plasma extractor FK-01 (Leadcore, Russia) and 200 ml of saline was added into the bag which was centrifuged at 1300 rpm for 10 min at 10°C and the supernatant was expressed out of the bag so that the remaining solution in the bag (30 ml) contained leucoconcentrate (WBC - 45.56 ± 23.93 × 106/ml and RBC - 1.76 ± 3.33 × 109/ml). Transduction of WBC with chimeric adenoviral vector (Ad5/35) carrying GFP gene was performed in the plastic bag with MOI 5 according to the count of WBC in the leucoconcentrate. After transduction for 12 hours, 200 ml of saline was added to the bag with leucoconcentrate, the mixture was centrifuged at 1000 rpm for 10 min at 10°C and the supernatant was squeezed out of the bag. The remained in the bag solution (30 ml) was considered as gene-modified leucoconcentrate carrying GFP gen (WBC - 22.63 ± 8.90 × 106/ml and RBC - 1.77 ± 1.21 × 109/ml). For in vitro study of GFP gene expression the samples of GML-GFP were cultivated for 60 hours after GML-GFP preparation. Fluorescent microscopy in the cytoplasm of the transduced WBC showed specific intensive green fluorescence. Flow cytometry analysis demonstrated that 2.5% of WBC from the GML-GFP efficiently expressed GFP. Thus leucoconcentrate after 72 h of transduction with Ad5/35-GFP with MOI 5 resulted in 2.5% of the GFP-positive cells. Thus the results of this study represent a simple, safe and effective approach for preparation of GML for personalized ex vivo gene therapy aimed at temporary production of the specific recombinant biologically active molecules for pathogenetic therapy of the varied nosological form, such as trauma, ischemic, degenerative, autoimmune, infection and other diseases. This study was supported by the grant of Russian Science Foundation 19-75-10030. Disclosures No relevant conflicts of interest to declare.

Surviving and Thriving Immediate Jeopardy in Infection Control from the Centers for Medicare and Medicaid

Background: Because of a patient death from a blood transfusion, a large hospital in Houston, Texas, underwent one of the largest unannounced CMS surveys in 2019. Methods: A 520-bed quaternary-care hospital was surveyed in one of the nation’s largest CMS surveys in March 2019, with a resurvey in June 2019. In an anticipated but unannounced arrival, ∼30 CMS surveyors evaluated the hospital and 10 Clinical Laboratory Improvement Amendments surveyors looked at the laboratory. They stayed for 11 consecutive days in March. On day 4, they declared that the hospital was in immediate jeopardy in infection control for the same observations noted by several surveyors. In addition, 11 CMS surveyors returned for a shorter resurvey in June. Results: The following 14 issues were listed under the infection control heading during the first survey, which led to the immediate jeopardy designation. The hospital’s infection prevention department committed to putting remediation processes, procedures, and audits in place during the first survey, which led to lifting the IJ before the surveyors left. The following shortcomings were recorded:(1)Inappropriate donning and doffing of personal protective equipment (PPE) for patients in isolationStandardized donning and doffing processes of PPE developed to include train-the-trainer and return demonstrations from >4,000 employees and providers followed by a minimum of fifty (50) audits/week with the goal of achieving 100% proper PPE donning and doffing for a minimum of three months, followed by a minimum of fifty (50) quarterly observations.(2)Environment Service (EVS) cleaning issues in isolation roomsTwo-person isolation room cleaning process developed, implemented, and audited a minimum of ten (10) times/week.(3)Incorrect set-up of dialysis machinesMinimum of five (5) dialysis machine set-ups audited/week.(4)Biohazard trash left in dialysis room between patientsMinimum random audits twice/week to look for biohazard trash.(5)Need for maintenance and cleanliness in the operating rooms (OR)Minimum three times/week audits of rotating ORs in all locations.(6)Rust noted on OR equipmentMinimum of twice/week audits looking for rust on OR equipment.(7)Insects noted in ORObservations for living insects will be audited twice/week.(8)Improper cleaning and high-level disinfection (HLD) of transvaginal probesMinimum of three times/week, cleaning and HLD processes of probes will be observed.(9)Matching patient to probes in their medical records needed clarificationMinimum of twice/week, logs will be audited to check that appropriate patient/probe linkage occurs.(10)Contaminated gloves used on a blood bag in ambulatory settingOnce/month, removal of blood bag from transport container will be observed to observe clean/dirty glove use(11)Lack of cleaning between patients of durable medical equipmentCleaning of DME will be observed for thoroughness a minimum of three times/week.(12)Sanitation and mislabeling issues in the kitchenA minimum of one (1) complete audit and two (2) abbreviated audits of kitchen sanitation and food labeling will be conducted per week.(13)Endoscopy misuse of test stripsTest strip audits showing appropriate labeling and use will be auditing a minimum of twice/week.(14)Process of air blowing of automatic endoscopic reprocessor (AER) needed improvement.Funding: NoneDisclosures: None