Journal of Haematology and Stem Cell Research <p><em>Journal of Haematology and Stem Cell Research (JHSCR)</em> is an official journal of Pakistan Society of Haematology, recognized by <strong>Higher Education commission of Pakistan in Y category)</strong>. It is a double blind double peer-reviewed being published biannual journal that delivers the premier quality peer-reviewed open access original research articles, reviews, and scholarly comment on pioneering efforts and innovative studies in the medical treatment of blood disorders. The journal has a strong clinical and pharmacological focus and is aimed at an international audience of clinicians and researchers in hematology and related disciplines, providing an online forum for rapid dissemination of recent research and perspectives in this area.</p> Pakistan Society of Haematology en-US Journal of Haematology and Stem Cell Research 2790-2277 <p><a href="" rel="license"><img src="" alt="Creative Commons License" /></a><br />This work is licensed under a <a href="" rel="license">Creative Commons Attribution-NonCommercial 4.0 International License</a>.<br />The licensor permits others to copy, distribute, display, and perform the work, as well as make and distribute derivative works based on it. The licensor permits others to copy, distribute, display, and perform the work for non-commercial purposes only.</p> CRISPR/Cas9 Gene Editing in Bacteria: Leading to Hematopoietic Stem Cell Editing in Pakistan <p><strong>Background:</strong> Deletion of LacZ gene in E. coli HB101-pBRKan using Clustered Regularly Inter-Spaced Short Palindromic Repeats-Cas9 (CRISPR/Cas9) gene-editing tool.</p> <p><strong>Methodology:</strong> Using BioRadTM USA “Out of the blue” gene editing kit, we demonstrated in vitro knock-out model for deletion of LacZ gene in E. coli HB101-pBRKan by employing CRISPR/Cas9 gene-editing tool. We followed an already designed modular plasmid assembly strategy and tested a single nuclease. Editing genomic locus was achieved within three days of minimal lab work.</p> <p><strong>Results: </strong>We were successful in gene editing of the E. coli LacZ gene using CRISPR/Cas9.</p> <p><strong>Conclusion:</strong> With the practical experience gained, we are confident enough to experiment in animal models to tackle the hematological disease.</p> Mohammad Abdul Naeem Copyright (c) 2022 Journal of Haematology and Stem Cell Research 2022-02-13 2022-02-13 2 1 4 7 A Study of Blood Donor Deferral Causes: Pre vs Post Donation and Transfusion Transmissible Infections <p><strong>Objectives: </strong>The aim of this study was to evaluate the prevalence of different causes of donor rejection before and after blood donation based on donor selection tools, and to see the usefulness of Chemiluminescence Immunoassay (CLIA) technique over rational screening by kit method in detecting borderline cases of TTI.</p> <p><strong>Methodology: </strong>All donors who came for the donation of blood over a period of 02 years between March 2019 to February 2021 were evaluated after taking their consent and seeking approval from Hospital Ethical Review Board.&nbsp;The data was analyzed to determine using SPSS version 20.</p> <p><strong>Results: </strong>Among 1849 individuals, mostly ranging between 25 to 45 years, presenting for blood donation, 164 (8.8%) were deferred. Among total deferred cases, 95(57.9%) were temporary deferrals and 69(42%) were permanent deferrals. The most frequent cause of deferral was low hemoglobin. Anemia was the leading cause of deferral in approximately one third of the rejected individuals (n&nbsp;= 51, 31%). One striking finding observed in this study was that 4.7% HCV positive and 62.5% HIV positive donors were in window period, which showed borderline results on CLIA.</p> <p><strong>Conclusion: </strong>Donor deferral, both pre and post blood donation&nbsp;is a major problem in our country and accounts for 8.8% of all prospective&nbsp;blood donors. Our results put an emphasis on addressing these causes efficiently to ensure safety of both the donor and potential recipients.</p> <p><strong>Keywords:</strong>&nbsp;Blood donors; Deferral; Anemia; Transfusion transmissible infections; Screening; Donor questionnaire.</p> Farah Hanif Sundas Ali Muhammad Bilal Habib Ayesha Siddiqa4 Muhammad Anees Copyright (c) 2022 Journal of Haematology and Stem Cell Research 2022-02-13 2022-02-13 2 1 13 18 Indirect Genetic Analysis of Pakistani Hemophilia a Pedigrees by using BclI and HindIII Polymorphic Markers <p><strong>Objective:</strong> The current study was aimed at analyzing the utility of intragenic polymorphic restriction sites (<em>Hind</em>III and <em>Bcl</em>I) in screening of haemophilia A patients and their immediate family members.</p> <p><strong>Methodology:</strong> Thirteen families having one or more affected child were recruited from Pakistan Institute of Medical Sciences. For Genomic DNA extraction, phenol-chloroform method was used. Genomic regions containing the RFLP site were amplified by using specific primers designer from flanking regions. The PCR products were digested using <em>Hind</em>III and <em>Bcl</em>I and the products were resolved on 6% Polyacrylamide gel electrophoresis and visualized by silver staining technique.</p> <p><strong>Results: </strong>A total of 100 X-chromosomes were investigated in this study (36 males, 32 females). The minor allele frequency for Intron 18/<em>Bcl</em>I T&gt;A was observed to be 0.68 while for Intron 19/<em>HindIII</em> C&gt;T it was observed to be 0.55 in our study population. The Intron 18/<em>Bcl</em>I T&gt;A marker was informative in four of the families while Intron 19/<em>Hind</em>III C&gt;T was informative for 8 families. The PIC was calculated to be 0.34 for <em>Bcl</em>I marker and 0.37 for <em>Hind</em>III .</p> <p><strong>Conclusion:</strong> The rate of heterozygosity and the PIC values obtained for the two markers suggests that <em>Hind</em>III marker is more informative as compared to <em>Bcl</em>I in our study population. The two markers were decisive in haplotyping 8 (66.6%) families.</p> Memoona Rasheed Naghmi Asif Asfa Zawar Khalid Hassan Tazheen Anwar Copyright (c) 2022 Journal of Haematology and Stem Cell Research 2022-02-13 2022-02-13 2 1 8 12 Association of Various Etiological Factors with Idiopathic Acquired Aplastic Anemia <p><strong>Objective:</strong> To determine demographic features, clinical features and association of various etiological factors in patients of idiopathic acquired aplastic anemia in local population at the time of diagnosis.</p> <p><strong>Patients and Methods:</strong> This cross sectional study was conducted in Department of Haematology, Pakistan Institute of Medical Sciences Islamabad and Department of Genetics, Children Hospital Lahore, from June 2015 to July 2017. Total 64 cases of peripheral blood pancytopenia having clinical suspicion of acquired aplastic anemia were enrolled in the study. Patients with congenital aplastic anaemia, patients with features of bone marrow dysplasia and abnormal infiltrates and patients with post chemotherapy and radiotherapy aplasia were identified and excluded from the study. Sixty patients diagnosed to have acquired aplastic anaemia on the basis of clinical features, peripheral blood and bone marrow findings, were included in this study as per inclusion criteria.</p> <p><strong>Results:</strong> Among 60 diagnosed cases of acquired aplastic anaemia, 34 were male and 26 were female with a male to female ratio of 1:1.30. Age distribution of study participants ranged from 1 to 84 years with a median age of 10 years. Most of the patients (73.3%) presented with non severe stage of disease. Different etiological factors reported to be implicated in acquired aplastic anaemia in the patients of this study were Viral infections (hepatitis B &amp; hepatitis C), Mycobacterial infection, Fertilizers &amp; Pesticides, Chemicals &amp; Toxins, Ionizing radiations, Autoimmune diseases (Rheumatoid arthritis &amp; Systemic lupus erythematosus), Transfusion associated GVHD, Pregnancy, Drugs (Analgesics &amp; Hypoglycemics) and Cigarette smoking. In 14 out of 60 patients (23.3%) various etiological factors were found to be associated with idiopathic acquired aplastic anaemia.</p> <p><strong>Conclusion:</strong> Aplastic anaemia was seen in patients of all ages and both genders were equally affected. Non severe aplastic anaemia was most common stage of the disease. Pallor, bleeding and fever were common clinical features. Etiological factors could not be identified in majority of the patients, however various etiological factors have been reported to be associated with increased risk of acquired aplastic anaemia. Identification and treatment of underlying cause may be beneficial in the management of patients with acquired AA. It may also guides in prevention of the disease.</p> Imran Khalid Farhan Abbas Zeeshan Mustafa Muhammad Kamran Copyright (c) 2022 Journal of Haematology and Stem Cell Research 2022-02-13 2022-02-13 2 1 19 25 Splenectomy in Patients with Beta Thalassaemia Major <p><strong>Objective: </strong>To evaluate the effect of splenectomy, in patients with beta thalassaemia major, on blood transfusion rates and blood counts<strong> &nbsp;</strong></p> <p><strong>Methods:</strong> In this prospective descriptive study patients with beta thalassaemia major, who underwent splenectomy were included . Inclusion criteria was thalasseamics&nbsp; with signs of growth retardation , poor health, leucopenia, thrombocytopenia, increased transfusion demand (more than 250 ml/kg/year or more than four transfusions per month) or respiratory embarrassment (mechanical discomfort) from massive splenomegaly even with lesser transfusion requirements&nbsp; were included.&nbsp; The transfusion requirement, pre- and post- splenectomy, was expressed as the volume of packed red cells used for transfusion in one year divided by the weight of the patient in mid year and expressed as ml/kg/year. Hypersplenism was established in patients having splenomegaly with accompanying anaemia, leucopenia, thrombocytopenia&nbsp; or any combination of these with hypercellular bone marrow .</p> <p><strong>Results: </strong>Majority were male (60.65%).Average hospital stay was six days. Pre-splenectmy, majority of patients had transfusion requirements more than 350 ml/kg/l. Blood trnasfusion requirements revealed a significant fall( less&nbsp; than 250 ml/kg/l) post splenectomy.One year follow up showed significant fall in transfusion rate and significant increase in haemoglobin . In&nbsp; 23.94%&nbsp; patients splenectomy was performed at an age less than ten years.</p> <p><strong>Conclusion : </strong>Splenectomy has a positive outcome on blood transfusion&nbsp; rates and haemoglobin&nbsp; levels in patients with beta thalassaemia major</p> Tazeen Anwar Gul-e-Najaf Bilal Ahmed Faheem Ahmad Subhani Copyright (c) 2022 Journal of Haematology and Stem Cell Research 2022-02-13 2022-02-13 2 1 26 30 The Hematological parameters in Malaria and its correlation with parasite Index: A cross-sectional study in Pakistan <p><strong>Objective</strong>: To determine the haematological parameters in malaria (Hb, TLC, Platelets) and correlate them with parasite index.</p> <p><strong>Methodology:</strong> This was a descriptive cross-sectional study conducted at the Fauji Foundation Hospital, Rawalpindi in the setting of the Pathology Department from March-September 2020. A total of 85 cases were included in the study. The sample size was calculated keeping confidence level=95%, SD= 0.35, absolute precision required= 4%. Non-probability consecutive sampling was used for data collection.</p> <p><strong>Results:</strong> The average age (years) of patients was 27.96+18.60. There were 213 (66.8%) male and106 (33.2%) female cases. Patients presented with average TLC and Hb levels as 5.92+2.22 and 11.22+1.68 respectively. Mean values of TLC among high and low parasite index (PI) were 6.14+1.67 and 5.83+2.41 respectively (p-value 0.564) whereas the mean values of Hb among high and low parasite index (PI) was 11.14+1.16 and 11.25+1.86 respectively (p-value 0.779). Mean platelet count among high and low parasite index (PI) was 27.44+90.81 and 61.20+31.09 respectively which was statistically significant (p-value 0.000).</p> <p><strong>Conclusion:</strong> Mild anaemia and moderate thrombocytopaenia were associated with high parasite index whereas no significant change was observed in TLC with high or low parasite index.</p> <p><strong>Keywords:</strong> Haematological parameters, Malaria, Parasite index, Pakistan.</p> Mariam Qamar Khan Prof. Dr. Lubna Zafar Dr Amina Zulfiqar Dr. Atif Abbas Dr Hassan Salim Amir Afzal Copyright (c) 2022 Journal of Haematology and Stem Cell Research 2022-02-13 2022-02-13 2 1 31 34 Compact introduction of stem cell association with skin cancer <p>Stem cells are defined as specific undifferentiated cells with self-replicating ability which are common to multicellular organisms. They possess the characteristics to replace, generate and endure terminally differentiated cells.Normal adult stem cells reside in most somatic tissues, where they form the cellular basis for tissue homeostasis, maintenance, and repair, as has been shown, for example, in the skin.It has been recently suggested that oncogenic mutations and other genetic and epigenetic defects could be inherited from transformed normal stem cells, giving rise to some of the cell populations observed in many tumors . A number of hypotheses, mostly based on new experimental evidences, have been proposed to explain the presence of cancer stem cells in cancer and the implications for future therapies.</p> Nida Sadaqat Copyright (c) 2022 Journal of Haematology and Stem Cell Research 2022-02-13 2022-02-13 2 1 35 44 A Review of Procurement Practices of Regional Blood Centres for Complex Equipment and Consumables <p>Access to accurate, safe, and adequate diagnostics and dependable laboratory services is vital for diagnosis, surveillance, and ensuring a safe supply of blood. However, testing results are only of value to those making clinical conclusions if the test results reported are well-timed and correct. Selection and procurement of diagnostics and laboratory technologies are time and again challenging due to the extensive choice of products and suppliers in the global market. A study was commissioned to review the current procurement practices for complex equipment and consumables to enable Regional Blood Centres to weigh up the advantages and disadvantages of the "purchase" and the "lease" model and to make an informed decision between both models. The study is expected to provide practical guidance on negotiation strategies with vendors and describe the procedures that Health Departments must follow to lease equipment or procure consumables for purchased equipment. The study concluded that the rental model is already implemented successfully in all provinces at various blood banks and laboratories. Reagent rental and cost-per-test contracts with diagnostic manufacturers provide a viable solution to the challenges of high capital investments saving a huge amount of financial resources.</p> Usman Waheed Noore Saba Saeed Ahmed Muhammad Nisar Copyright (c) 2022 Journal of Haematology and Stem Cell Research 2022-02-13 2022-02-13 2 1 45 48 Emerging Horizons in Haemophilia Care Nadeem Ikram Tahira Zafar Copyright (c) 2022 Journal of Haematology and Stem Cell Research 2022-02-13 2022-02-13 2 1 1 3