Prevalence of A2 and A2B Subgroups among Blood Groups A and AB in healthy donors.

Objective: To determine the frequency of A2 and A2B subgroups among blood groups A and AB in healthy donors. Methods: It was a Cross-Sectional study, conducted at the Department of Hematology & Transfusion Medicine, UCHS, The Children's Hospital Lahore and Sundas foundation Lahore from June 2022 to December 2022 including 13,120 healthy blood donors of both genders, after taking informed consent. Venous blood samples of donors were collected in EDTA vials (3ml) and serum gel vial for routine blood grouping which was done by standard tube method. Further testing of donors positive for an antigen (blood Group-A and AB) was performed using anti-A1 lectin by standard tube method as per manufacturer's instruction. The data was analyzed using SPSS version 23. Results: Among 13120 blood donors, 12857 (97.9%) were male and 263 (2.0%) were female with mean age of 36.7 years ± 15.04 years. Majority of them (91.7%) were of Punjabi ethnicity. Donors having blood group phenotype A and AB were 3890 (29.6%). Among blood Group-A donors, A1 was found in 97.8% and A2 in 2.2% donors. While among Blood Group-AB, 96.7% donors belonged to A1B blood group and 3.2% belonged to A2B blood group. Conclusions: Blood group A2 and A2B do exist in blood donors of Punjabi ethnicity. The knowledge of presence of these blood groups' phenotypes in our population can provide a better base for transfusion staff to do troubleshooting in compatibility testing and to avoid any rare but hazardous transfusion outcome.

Complications of Therapeutic Plasma Exchange in pediatric patients: An experience at a tertiary care hospital.

Objective: To find the incidence of various complications of therapeutic plasma exchange (TPE) in ICU admitted children and to determine their association with age, gender, blood group and diagnosis of the patients. Methods: In this observational study, data of 24 patients who underwent 125 sessions of TPE was collected from the Pediatric Intensive care unit (PICU) and Hematology department of The Children's Hospital, Lahore from December 2020 to November 2021. Age, gender, blood group, indications and complications observed during and after the TPE procedure were documented on a pre-designed proforma. The data was analyzed by using SPSS version 23. Quantitative variables were presented in the form of mean and standard deviation. Qualitative variables like gender, blood groups, indications and complications of plasmapheresis were presented as frequency and percentage. Chi square test was applied for comparison of variables. Results: Among the 24 patients, 45.8% were of age group five to ten years with mean age of 7.58 years± 2.04 years and male to female ratio of 0.84:1. Guillain-Barré syndrome (GBS) and Neuromyelitis Optica spectrum disorder (NMO-SD) were the most prevalent among the patients who underwent TPE. Most common complication was hypotension (44.9%), others were febrile reactions (11.6%), unstable vital signs (14.5%) and allergic reactions (24.6%). Blood group, clinical condition and diagnosis of the patient showed significant association with the incidence of TPE related complications. Conclusion: The majority of problems caused by TPE are considered to be minor. Sudden fall in blood pressure, pruritus, urticarial rash and fever are the common adverse consequences among pediatric patients. Blood group and diagnosis of the patient can determine the development of such complications during plasmapheresis procedure.

Comparison of single and double centrifugation methods for preparation of Platelet-Rich Plasma (PRP).

Objective: To compare the platelet count, platelet concentration/yield, residual Red blood cells (RBCs) and White blood cells (WBCs) counts in platelet-rich plasma (PRP) samples prepared from the single- and the double-centrifugation protocols. Methods: It was a Cross-Sectional study, conducted at the Department of Hematology & Transfusion Medicine, The Children's Hospital and UCHS, Lahore from October 2021 to January 2022 including 50 voluntary, healthy individuals of age 20-45 years of both genders, after taking informed consent. Complete blood count analysis of all participants was done initially by drawing 3ml blood in EDTA vial. From all the participants, 20 ml venous blood sample was taken in syringes containing tri-sodium citrate and then shifted to harvest tubes. Group-I comprised of PRP samples prepared by single- centrifugation method. While Group-II samples were prepared by Double-centrifugation method consisting of soft and hard spin. The platelet, RBC and WBC counts in prepared PRP samples were determined by using automated SYSMEX XP-100 hematology analyzer. Platelet yield or Platelet concentration (%) was calculated for samples using formula. The data analysis was done using SPSS version 23. Results: The mean PRP platelet count in Group-I was 594.6±157.4×103/µl whereas in Group-II was 923.06 ± 127.58×103/µl. In Group-I, the mean platelet concentration/yield in PRP was 175.75 ± 55.08% while in Group-II, it was 276.78 ± 112.7%. Significant difference was observed between the platelet counts and platelet concentration/yields from the PRP samples of two Group-s (p < 0.01). Significant difference between the WBCs count was also observed (p < 0.01) with higher WBCs in Group- I PRP. Residual RBCs were almost same among two Group-s. Conclusions: The double centrifugation protocol resulted in higher platelet quantity and yield with less contamination by red and white blood cells than did the single centrifugation protocol for PRP preparation. So, double centrifugation method is beneficial in preparation of autologous as well as allogenic PRP.

Effect of donor and red blood cells concentrate characteristics on recipient hemoglobin increment following red blood cells transfusion in pediatric patients.

Objectives: To determine the effects of donor and red blood cells concentrate characteristics on recipient hemoglobin increment following red blood cells transfusion in pediatric patients. Methods: This cross-sectional study was conducted at The Hematology & Transfusion Medicine Department of The UCHS & The Children's Hospital, Lahore from 23rd December 2020 to 31st July 2021 after Institutional Ethical committee approval. After taking informed consent from parents/guardians, One hundred recipients receiving RBCs unit transfusion studied along with the respective donors. The donor's details were recorded on a pre-designed proforma which included age, gender, Body Mass Index (BMI), CBC analysis (Hemoglobin [Hb] & Hematocrit) and blood group. Components' preparation, storage and modifications details were also recorded. Hb levels of recipient were determined 12 hours prior to transfusion and 12-18 hours after transfusion. The data was analyzed on SPSS version 26. Results: Among recipients, the mean age was 5.25 ±3 years and male to female ratio was 1.16:1. The mean pre-transfusion Hb level of patients was 6.48g/dl (SD: 2.15) and mean post transfusion Hb was 8.824 g/dl (SD: 2.03) with a significant raise after transfusion (p< 0.001). Majority donors (60%) were between 18 to 30 years of age and mean age was 30.7 years (SD: 9.04). The hemoglobin increment was reduced for transfusion of RBC units from donor with greater age. Post- transfusion Hb rise was more in Rh D positive donations than Rh D negative (p< 0.0001). No significance of donors' gender, BMI, Hb and hematocrit was found in relation to Hb increment. Among RBCs concentrate features, washing with normal saline found to have greater Hb increment, particularly in Thalassemia patients (p< 0.0001). Conclusion: Donors' age and Rh blood group and red blood cell concentrates' washing accounts for significant rise in recipient's post-transfusion hemoglobin. These factors may be used to predict changes in recipients' hemoglobin before transfusion.

Utility of the ISTH bleeding assessment tool (BAT) in diagnosis of Glanzmann Thrombasthenia patients.

Objectives: To assess the utility of ISTH-BAT (International Society on Thrombosis and Hemostasis- Bleeding Assessment Tool) in the diagnosis of Glanzmann Thrombasthenia (GT) in comparison to controls. Methods: It was a case-control study carried out at The Children's Hospital, Lahore from January 2012 to May 2021. All patients from neonates to 18 years with a final diagnosis of GT were studied retrospectively. The clinical details were collected from hospital records and telephonically on ISTH-BAT questionnaire after taking informed consent. The same proforma was obtained from 75 healthy controls. Data was analyzed on SPSS version 26. Results: Out of 427 patients with suspected platelet function disorders, 133 were diagnosed as GT. The mean age was 7.29±5 years. Male to female ratio was 1.1:1. Among cases, 76.6% were products of consanguineous marriage. Epistaxis was the commonest symptom with highest score (p value<0.001). Cutaneous and oral cavity bleeds were more severe and frequent in patients than controls (p value < 0.004). The median ISTH-BAT score among patients was nine while in control group was one. Sensitivity was 86.4%, specificity was 77.3%, positive predictive value was 0.87 and negative predictive value was 0.76. Area under the receiver operator curve was 0.78 (95% confidence interval 0.82-0.90, p< 0.001*). Conclusion: ISTH-BAT scores were significantly higher in GT patients than controls. So, we recommend the inclusion of ISTH-BAT in diagnostic evaluation of patients with suspected Glanzmann Thrombasthenia.

T lymphoblastic lymphoma with BCR-ABL negative chronic myeloid leukaemia: a novel association.

Lymphoblastic lymphoma and chronic myeloid leukaemia (CML) are two distinct neoplasms with different pathogenesis and clinical presentation. We hereby share a challenging case of a child presenting with fever, leucocytosis, generalised lymphadenopathy and massive splenomegaly. He was diagnosed as having novel association of concurrent T-lymphoblastic lymphoma diagnosed on cervical lymph node biopsy with BCR-ABL negative CML on bone marrow aspirate. The study of more such cases is needed for optimal patient management.

Is leukopenia and lymphopenia a characteristic feature of COVID-19 in children?

OBJECTIVES: To analyze whether leucopenia and lymphopenia a characteristic feature of children with COVID-19 and to find out its association with the disease severity. METHODS: This was a descriptive cross-sectional study conducted at The Children's Hospital Lahore from March 2020 to October 2020. All confirmed cases of COVID-19 infection and post-COVID MIS-C/Kawasaki Disease diagnosed on the basis of RT-PCR and Antibody test respectively were included. Complete blood and differential counts were performed on the day of admission. RESULTS: Out of a total of 83 patients 60 (72%) were diagnosed as COVID-19 and 23 (28%) as post-COVID MIS-C/KD. The mean age of children was 7.0±4.3 years (95%CI: 6.07 - 8.75) with a male preponderance 51 (61%). Twenty (24%) children had an underlying comorbidity and 7 (8%) were surgical cases. Our case fatality rate was 5 (6%) and all children who died had an underlying comorbid condition. In both, COVID and MIS-C/KD the mean leukocyte count was (14.0 ± 12.5 vs 13.6 ± 6.9 x109/L), respectively (p=0.888). The mean lymphocyte count in children with COVID was (39.1 ± 21.4%). Patients with MIS-C/KD showed significantly higher levels of neutrophil count (76.5 ± 15.0%) as compared to children with COVID (52.0 ± 22.1%), absolute lymphocyte count was (5.02±4.81 vs 2.13±0.95 x109/L) in COVID and MIS-C respectively (p=<0.001). In 60 COVID-19 patients, the mean neutrophil lymphocyte ratio (NLR) in mild-moderate and severe-critical group was 2.00 and 5.08 respectively (p=0.009). CONCLUSION: The blood picture of COVID-19 in children does not show leukopenia. NLR was a prognostic factor to assess the severity in COVID-19 patients. The presence of an underlying comorbid conditions is significant a risk factor for poor outcome.

Characterization of the mutation spectrum in a Pakistani cohort of type 3 von Willebrand disease.

INTRODUCTION: Type 3 von Willebrand disease (VWD), a severe autosomal recessive hereditary bleeding disorder, is described by the virtual absence of von Willebrand factor (VWF). In consanguineous populations, for example Pakistan, the disease is reported with a higher incidence rate than the worldwide prevalence. AIMS: This study aims to characterize molecular pathology and clinical profile of type 3 VWD cohort of Pakistani origin. METHODS: In total, 48 patients were enrolled in the current study. Initially, the index patients (IPs) were evaluated by a standardized questionnaire for recording bleeding manifestations and by performing conventional coagulation tests. The diagnosis of VWD type 3 was confirmed by VWF antigens less than 5 IU/dL. Direct sequencing of VWF gene (VWF) was carried out to identify causative gene variations. We evaluated the potential consequence of novel splice site and missense variations by predictive computational programs and in silico structural analysis. RESULTS: VWF mutations were detected in 46 out of 48 IPs (95.8%), predominantly as homozygous variants. In total, twenty-nine different gene defects were characterized in this cohort from which 10 (34.5%) are novel. The majority of the mutations were null alleles (66%; including gene conversions, nonsense, splice site variations, small deletions and insertions), and 34% of them were missense substitutions. CONCLUSION: Herein, we reported for the first time, the pattern of gene defects in Pakistani type 3 VWD cohort. We identified a wide heterogeneous mutation spectrum along with variability in the type of bleeding episodes.

Correction to: Identification of novel mutations in congenital afibrinogenemia patients and molecular modeling of missense mutations in Pakistani population.

[This corrects the article DOI: 10.1186/s12959-017-0143-3.].

Identification of novel mutations in congenital afibrinogenemia patients and molecular modeling of missense mutations in Pakistani population.

BACKGROUND: Congenital afibrinogenemia (OMIM #202400) is a rare coagulation disorder that was first described in 1920. It is transmitted as an autosomal recessive trait that is characterized by absent levels of fibrinogen (factor I) in plasma. Consanguinity in Pakistan and its neighboring countries has resulted in a higher number of cases of congenital fibrinogen deficiency in their respective populations. This study focused on the detection of mutations in fibrinogen genes using DNA sequencing and molecular modeling of missense mutations in all three genes [Fibrinogen gene alpha (FGA), beta (FGB) and gamma (FGG)] in Pakistani patients. METHODS: This descriptive and cross sectional study was conducted in Karachi and Lahore and fully complied with the Declaration of Helsinki. Patients with fibrinogen deficiency were screened for mutations in the Fibrinogen alpha (FGA), beta (FGB) and gamma (FGG) genes by direct sequencing. Molecular modeling was performed to predict the putative structure functional impact of the missense mutations identified in this study. RESULTS: Ten patients had mutations in FGA followed by three mutations in FGB and three mutations in FGG, respectively. Twelve of these mutations were novel. The missense mutations were predicted to result in a loss of stability because they break ordered regions and cause clashes in the hydrophobic core of the protein. CONCLUSIONS: Congenital afibrinogenemia is a rapidly growing problem in regions where consanguinity is frequently practiced. This study illustrates that mutations in FGA are relatively more common in Pakistani patients and molecular modeling of the missense mutations has shown damaging protein structures which has profounding effect on phenotypic bleeding manifestations in these patients.

Atypical Chronic Myeloid Leukemia, BCR-ABL1 Negative, in a 5-Month Baby - ARare Presentation.

Atypical chronic myeloid leukemia (aCML) is a neoplasm with poor prognosis, characterized by myeloid hyperplasia, dysmyelopoiesis and absence of BCR-ABL1 gene. Clinically, the disease course may be similar to chronic myeloid leukemia (CML), BCR-ABL1 positive. It presents in seventh to eighth decade of life with few cases of paediatric aCML being reported. Here, we report a case of aCMLin a 5-month baby who presented with massive splenomegaly. The diagnosis was in accordance with the WHO criteria established in 2008.

Congenital amegakaryocytic thrombocytopenia with multiple physical anomalies in a female neonate.

Congenital Amegakaryocytic Thrombocytopenia (CAMT) is a rare disorder of infancy characterized by isolated thrombocytopenia along with hypoplasia or aplasia of megakaryocytes in the bone marrow. It is caused by c-mpl mutation which disrupts the function of thrombopoietin (TPO) receptor. CAMT in association with physical anomalies is a rare entity with only limited data from single case reports being available. Here we present a case of 35 days neonate who had CAMT together with facial malformations and cardiac defects.