Umbilical cord blood quality and quantity: Collection up to transplantation Seyed Hadi Mousavi, Morteza Zarrabi, Saeid Abroun, Mona Ahmadipanah, Bahareh Abbaspanah Asian Journal of Transfusion Science 2019 13(2):79-89 Umbilical cord blood (UCB) is an attractive source of hematopoietic stem cells for transplantation in some blood disorders. One of the major factors that influence on transplantation fate is cord blood (CB) cell count, in addition to human leukocyte antigen similarity and CD34+ cell number. Here, we review the factors that could effect on quality and quantity of CBUs. Relevant English-language literatures were searched and retrieved from PubMed using the terms: CB, quality, collection, and transplantation. The numbers of total nucleated cells (TNCs) and CD34+ cells are good indicators of CB quality because they have been associated with engraftment; thereby, whatever the TNCs in a CB unit (CBU) are higher, more likely they led to successful engraftment. Many factors influence the quantity and quality of UCB units that collect after delivery. Some parameters are not in our hands, such as maternal and infant factors, and hence, we cannot change these. However, some other factors are in our authority, such as mode of collection, type and amount of anticoagulant, and time and temperature during collection to postthaw CBUs and freeze-and-thaw procedures. By optimizing the CB collection, we can improve the quantity and quality of UCB for storage and increase the likelihood of its use for transplantation.

Role of plasma exchange in postpartum microangiopathies: An experience from a tertiary care center Rekha Hans, Satya Prakash, Divjot Singh Lamba, Ratti Ram Sharma, Pankaj Malhotra, Vanita Suri, Neelam Marwaha Asian Journal of Transfusion Science 2019 13(2):90-94 BACKGROUND: Postpartum microangiopathies are rare but are associated with high maternal and fetal mortality requiring early diagnosis and prompt treatment to improve the outcome. AIMS AND OBJECTIVES: This retrospective study aims to assess the efficacy of plasma exchange (PE) therapy in postpartum thrombotic microangiopathies. MATERIALS AND METHODS: We did retrospective analysis of all plasma exchange procedures performed in patients of postpartum thrombotic microangiopathies over a period of 1 year (2015-2016). Patient's pre- and post-plasma exchange hematological and biochemical parameters were recorded and compared for analyzing the response to the therapy. Patients were followed telephonically even after their discharge from the hospital. RESULTS: Hematological and renal profile improved in 8 out of 9 patients after PE therapy. Survival after PE therapy was 40% in post partum atypical HUS and 75% in patients with HELLP syndrome at 4 months of follow up. CONCLUSION: Early initiation of PE therapy in postpartum thrombotic microangiopathies can reduce morbidity and mortality associated with them.

Comparison of UV spectrometry and fluorometry-based methods for quantification of cell-free DNA in red cell components Dheeraj Khetan, Nitesh Gupta, Rajendra Chaudhary, Jai Shankar Shukla Asian Journal of Transfusion Science 2019 13(2):95-99 BACKGROUND: Stress and shear force applied on blood components during processing and storage may induce cellular damage leading to release of cell-free DNA (cfDNA). In this study, we have compared ultraviolet (UV) spectrophotometry with UV-induced fluorescence for the quantification of cfDNA in red cell supernatant. MATERIALS AND METHODS: cfDNA was extracted from 200 μL sample of supernatants from 99 packed red blood cells using QIAamp DNA Blood Mini Kit (Qiagen, Germany). Quantification of cfDNA was done using two different methods: one based on spectrophotometry (NanoDrop 2000c, ThermoFisher Scientific, USA) and another based on fluorometry (Qubit 2.0, Life Technologies, ThermoFisher Scientific, USA). Interassay variability of both the methods was estimated using serial dilutions of standard with known DNA concentration. RESULTS: DNA quantification by both the methods was close to actual amount of known standard in dilutions with higher concentration of DNA (21.68 to 2.71 ng/μl). While at higher dilutions, quantification by NanoDrop was neither precise nor accurate. Median cfDNA concentration in the study units was found to be 1.60 ng/μl (25th–75th percentile range: 1.10–2.10) by UV spectrophotometry (NanoDrop) compared to 0.080 ng/μl (25th–75th percentile range: 0.050–0.130) by fluorometry (Qubit). CONCLUSION: Due to high interassay variability between the two methods and the better precision and accuracy of Qubit, it is recommended that fluorometry-based method be used for the quantification of cfDNA in blood components.

Reversal of warfarin-coagulopathy: How to improve plasma transfusion practice in a community hospital setting? Lubna Bashir Munshi, Braghadheeswar Thyagarajan, Aasems Jacob, Shil Patel, Steve Zheng Liu, Arpad Szallasi Asian Journal of Transfusion Science 2019 13(2):100-104 BACKGROUND: Plasma is often given inappropriately to reverse warfarin-induced coagulopathy, wasting health-care resources and exposing the patients to transfusion-associated risks. AIMS: The clinical practice at our institution was evaluated in order to reduce the number of unnecessary plasma transfusions. MATERIALS AND METHODS: Retrospective audit of plasma transfusions was done (July 2014 to June 2015). DESIGN: To improve the clinical practice, a two-prong strategy was implemented: (1) in-service was given to clinicians on the warfarin-reversal guidelines and (2) for a 30-day period, plasma orders were placed on the approval list of the Transfusion Medicine Service. RESULTS: Of the 729 units of plasma, 189 (26% of total) were given for the reversal of warfarin-induced coagulopathy. The medical charts of these patients were reviewed: 46 units of plasma (~25%) were given inappropriately (e.g., patients with minimally elevated international normalized ratio, no evidence of bleeding, and no surgery within 24 h). To check the effectiveness of our intervention, two audits of plasma transfusions were done. During the first audit (January 1–February 29, 2016), 24 patients received plasma to reverse warfarin-coagulopathy. Medical chart review revealed that the vast majority of plasma orders (96.66%) followed the guidelines. A second audit was carried out a year later (January 1–March 31, 2017): during this 3-month period, 47 patients were transfused with plasma for warfarin reversal with a 94% adherence to the guidelines. CONCLUSION: We conclude that plasma transfusion practices may be improved by a combination of education and active enforcement of warfarin reversal guidelines.

Mutational analysis of thalassemia in transfusion-dependent beta-thalassemia patients from central India Manisha Shrivastava, Rashmi Bathri, Nirupama Chatterjee Asian Journal of Transfusion Science 2019 13(2):105-109 BACKGROUND: Thalassemia and hemoglobin (Hb) disorders are the most common genetic disorders among humans. These disorders entail huge morbidity, economic, and psychological burden on the families of the affected. Genetic counseling and prenatal diagnosis are the steps, which helps to reduce this burden. At present, there is paucity of data on the mutational spectrum of thalassemia from the central Indian region. METHODS: Blood samples were collected from 62 transfusion-dependent patients, demographic and relevant data were collected and screened for the two rare mutations − 88 (C-T) and CAP + 1 (A-G) using amplification refractory mutation system-polymerase chain reaction (PCR) and GAP PCR technique. PCR was performed for rare Hb disorders such as Hb Lepore and δ β chain disorder by GAP PCR in addition to five common Indian beta-thalassemia mutations IVS1-5 (G-C), IVS1-1 (G-T), Cd41/42 (−TCTT), Cd8/9 (+G), 619 bp deletion. RESULTS: Overall 93.5% of the mutations could be identified. Among the abnormal Hb, sickle cell and HbE were found at 4% and 3% of all the loci studied. We also reported two loci with Hb δ β and one locus with Hb Lepore in the present samples. IVS I-5 (G–C) was the common mutation (46%) followed by IVS I-1 (G–T) (12%) and 619 bp (9%). CONCLUSION: The identification of the genotypes helps to define the severity of the phenotype, plan therapy and form the basis of the comprehensive diagnostic database that would be useful not only for genetic counseling but prenatal diagnosis as well, contributing to the current focus of the National Policy to prevent and control hemoglobinopathies.

Evaluation of molecular typing and serological methods in solving discrepant results of weak and partial D (Rh) in South Egypt Rania M Bakry, Eman Nasreldin, Ashraf E Hassaballa, Samar M Mansour, Sahar A Aboalia Asian Journal of Transfusion Science 2019 13(2):110-114 INTRODUCTION: Rh discrepancies produced by partial and weak D phenotypes are a problem during routine testing. Some blood units with weak and partial D expression may be missed by serology. Overcoming the limitations of serology can be achieved by molecular typing. Our objective was to evaluate currently used serologic methods with the molecular analysis in solving discrepant results of weak and partial D (Rh) in South Egypt. PATIENTS AND METHODS: Fifty blood donor and patient samples with undetermined D phenotype were subjected to serology to define their phenotype using identification (ID)-Card “ID-partial RhD typing set” using six monoclonal anti-D panels, followed by molecular typing using polymerase chain reaction sequence-specific primer kit. RESULTS: Molecular typing confirmed most of the serology results; two samples previously resolved as partial D Type 3 and DFR by serological methods were clarified by molecular techniques – one sample as weak Type 4 and the other sample as weak Type 3. Among the weak D alleles found in our study, Type 4 was the most common, with a frequency of 20%, followed by Type 3 (14%), Type 1 (8%), Type 2 (6%), and finally, Type 5 with a frequency of 3%. The most common types of partial D were partial D Type D5 (14%) and Type D3 (10%). CONCLUSION: Our study identified D variants (weak D and partial D categories) of the antigen D and determined the frequency and composition of partial D and weak D alleles in our population. Molecular typing also confirmed most of the results obtained from serological methods.

Comparison of a column agglutination technology-based automated immunohematology analyzer and a semiautomated system in pretransfusion testing Ravi C Dara, Aseem Kumar Tiwari, Subhasis Mitra, Deviprasad Acharya, Geet Aggarwal, Dinesh Arora, Gunjan Bhardwaj Asian Journal of Transfusion Science 2019 13(2):115-119 INTRODUCTION: Semi-automated equipment using Column agglutination technology (CAT) is widely used where centrifugation and incubation are automated but substantial amount of the work is still executed manually. Larger laboratories are moving towards automation to eliminate errors, reducing exposure to bio-hazardous samples, assuring traceability, reliability, turnaround time (TAT) and throughput. Moving towards automation and greater reliability, we therefore, decided to install an automated immunohematological (IH) analyzer “Vision”. In this study we evaluated reliability and performance before clearing “Vision” for routine use. MATERIALS AND METHODS: Study was conducted in the Department of Transfusion-Medicine. The primary objective was to assess the reliability of results and compare with routine use semi-automated BIOVUE system (Reference system). Secondary objective was to evaluate the performance (TAT and throughput) of the Vision to handle routine and emergency workload. RESULTS: Total of 1276 known samples were used to assess 2640 pre-transfusion tests (1229 ABO/Rh D typing; 1229 antibody screening; 54 antibody identification; 86 crossmatch and 42 DAT). All 1229 ABO Rh typing results were concordant between the two systems. Overall agreement between the Vision IH analyzer and reference system for ABO Rh typing was 99.95%. All antibody screening, crossmatch and DAT results were concordant between the two systems. TAT of Vision was substantially shorter than the reference system for all test profiles. CONCLUSION: Based on study results, Vision was approved for routine use in laboratory. It was found to be reliable with considerably shorter TAT and capable of handling high throughput of immunohematological tests.

Effectiveness of bone marrow-derived mononuclear stem cells for neurological recovery in participants with spinal cord injury: A randomized controlled trial Rajeshwar Nath Srivastava, Ashok Kumar Agrahari, Alka Singh, Tulika Chandra, Saloni Raj Asian Journal of Transfusion Science 2019 13(2):120-128 BACKGROUND: Complete lesion after spinal cord injury (SCI) remains irreversible with little hope of neurological recovery. Newer interventions such as re-stimulation of damaged neurons using artificial agents and the use of stem cells for neuronal regeneration have shown promising results. AIM: This study was undertaken for evaluating the neurological status of acute SCI participants after stem cell augmentation and comparing them with other treatment methods. SETTING AND DESIGN: Randomized controlled trial in the northern Indian population. MATERIALS AND METHODS: A total 193 SCI participants of complete paraplegia with unstable T4–L2 injury having thoracolumbar injury severity score ≥4 were enrolled in this study. Participants were randomly allocated for three different treatment modalities, namely, conventional with stem cell augmentation (Group-1), conventional (Group-2), and conservative (Group-3). Neurological recovery after 1 year was evaluated through the ASIA Impairment Scale (AIS)-grading, sensory, and motor scores. STATISTICAL ANALYSIS: T-test for sensory-motor score analysis of each group and analysis of variance for comparison of same variables between the groups. RESULTS: After 1-year significant difference was observed in the AIS-grade, sensory and motor scores in-Group 1 (P < 0.001). In Group-1 versus 2, the mean difference at 1 year for AIS grade, sensory and motor scores were 0.40 (P = 0.010, 95% confidence interval [CI] 0.075–0.727), 8.52 (P = 0.030, 95% CI 0.619–16.419), and 4.55(P = 0.003, 95% CI 1.282–7.815), respectively. In Group-1 versus 3, 1.03, 19.02 and 7.22 (P < 0.001 for each of the parameters) and in Group-2 versus 3, 0.63 (P < 0.001), 10.49 (P = 0.009), and 2.68 (P = 0.019), respectively. CONCLUSIONS: Significant motor neurological recovery and AIS-grade promotion was observed in Group-1 as compared to Group-2 and 3.

Detection of a rare subgroup of A phenotype while resolving ABO discrepancy Revathy Nair, Harita Gogri, Swati Kulkarni, Debasish Gupta Asian Journal of Transfusion Science 2019 13(2):129-131 Weaker subgroups of ABO blood group system give rise to discrepancies between forward and reverse grouping and cause diagnostic difficulties in routine blood banking. Weaker subgroups of A blood group that have been reported so far include A3, Aend, Ax, Am, Ay, and Ael. We report a case of a 54-year-old patient whose red cells showed a discrepancy between cell and serum grouping on initial testing. Serological investigation included absorption elution tests and saliva testing after performing initial blood grouping. Molecular genotyping of the ABO gene was performed by DNA sequencing of exons 6 and 7 of the ABO gene. The serological characteristics of the patient's red cells were similar to Ax subtype. The patient was a secretor and only H substance was present in the saliva. Serum did not show the presence of anti-A1. Molecular genotyping confirmed the ABO status as Aw06/O13. The weak A phenotype identified in the propositus had serological characteristics similar to Ax and showed the ABO genotype Aw06/O13. Although Aw06 allele has been previously reported in the Indian population, this is the first study to report O13 allele in the Indian population.

Deciphering a delayed hemolytic transfusion reactions nightmare – Case of Chido/Roger antibodies Soumya Das, PS Priyamvada, Abhishekh Basavarajegowda, Ankit Mathur Asian Journal of Transfusion Science 2019 13(2):132-135 Antibodies against Rh (CEce) and Kidd (Jka and Jkb) system antigens are mostly implicated in delayed hemolytic transfusion reactions (DHTR), which is a potentially life-threatening complication observed in patients receiving chronic transfusions. Here, we are describing a case of Chido/Roger antibody which presented to our laboratory as DHTR. The clinical presentation and laboratory findings including the immunohematological workups with regard to the reaction are discussed, with a special emphasis on the benefit of identifying such an antibody and obtaining blood unit for transfusion supports the patient with respect to providing a compatible unit.