Fibrin glue in managing intractable gingival bleeding in patients with inherited bleeding disorders-a quasi-experimental pilot study.

This study presents the clinical outcomes of using inhouse prepared fibrin glue for controlling gingival bleeding in patients with inherited bleeding disorders (IBD). The objective of the study was to assess the reduction in transfusion days and improvement in compliance for dental evaluation over a one-year period in a low-to-middle-income country. The quasiexperimental pilot study included 40 IBD patients with gingival bleeding. These were divided into two groups: Group A received fibrin glue (n=20), while Group B did not (n=20). The study compared outcome metrics, including the number of treatment days and blood components transfused, using non-parametric tests with a significance threshold of p<0.05. Results showed that Group A required fewer blood components (n=154) as compared to Group B (n=204) (p<0.001). Patients in Group A with Glanzmann thrombasthenia (GT) had a shorter treatment duration (one day) than those in group B (three days) (p<0.01). In conclusion, the application of fibrin glue effectively managed intractable gingival bleeding in IBD patients.

Recurrent pregnancy loss in a patient with anti-Rh17.

BACKGROUND: Rh is one of the most important blood group systems in transfusion medicine. The two homologous genes RHD and RHCE are located on chromosome 1p36.11 and encode for RhD and RhCE proteins, respectively. Complex genetic polymorphisms result in a variety of antigenic expression of D, C, E, c, and e. Here, we describe a case of a young female with D-- who developed anti-Rh17 secondary to blood transfusion and had signs of haemolytic disease of the fetus and fetal death in five consecutive pregnancies. CASE DESCRIPTION: EDTA-whole blood samples were collected from the patient, husband and eight siblings for blood grouping, phenotyping, and red cell antibody screening. Extracted DNA was genotyped by SNP-microarray and massively parallel sequencing (MPS) with targeted blood group exome sequencing. Copy number variation analysis was performed to identify structural variants in the RHD and RHCE. Routine phenotyping showed all family members were D+. The patient's red blood cells were C-E-c-e-, Rh17- and Rh46- and had anti-Rh17 and anti-e antibodies. MPS showed the patient carried a wildtype RHD sequence and homozygous for RHCE (1)-D (2-9)-CE (10) hybrid gene predicted to express a D-- phenotype. CONCLUSIONS: Our patient had a rare D-- phenotype and confirmed to have RHCE/RHD hybrid gene with replacement of 2-9 exons of RHCE by RHD sequences. Unfortunately, our patient developed anti-Rh17 and anti-e antibodies due to blood transfusion and suffered fetal demise in her very first pregnancy. The adverse outcomes could have been prevented by active prenatal management.

Burden of ocular abnormalities in patients with beta thalassaemia: a cross-sectional study.

INTRODUCTION: In Pakistan, the reported carrier rate of thalassaemia is estimated to be 5%-8% with 5000 new patients diagnosed every year. Several known systemic complications of beta thalassaemia major have been studied, but no studies are conducted to assess ocular complications among these patients from our population. METHODS: It was a cross-sectional study design conducted at three private and public sector centres in Pakistan. We recruited 203, 11-17 years old children with beta thalassaemia major in our study. Frequency of overall ocular complications such as retinal pigment epithelium degeneration, visual field defects, increased retinal vascular tortuosity, lenticular opacities, anterior segment abnormality, etc among beta thalassaemia patients were verified by an ophthalmologist. RESULTS: On univariate analysis male gender (prevalence ratio (PR): 1.023 (0.903 to 1.160), OGTT levels (PR: 0.99 (0.978 to 1.003) and serum calcium levels (PR: 0.716 (0.616 to 0.936)) were significantly associated with ocular complications. However, on multivariable analysis after adjusting for covariates we observed that the prevalence of ocular complications was 88% higher in males as compared with females. Moreover, with every one unit increase in serum calcium levels the prevalence of ocular complications were decreased by 24%. CONCLUSION: Our study results showed that the frequency of ocular complications in beta thalassaemia children was 22.7%. Male gender was a risk factor for ocular complications among children with beta thalassaemia. However, high calcium levels among these patients were found to be protective for ocular complications.

IgG antibodies to SARS-CoV-2 in asymptomatic blood donors at two time points in Karachi.

INTRODUCTION: An estimated 1.5 million cases were reported in Pakistan until 23 March, 2022. However, SARS-CoV-2 PCR testing capacity has been limited and the incidence of COVID-19 infections is unknown. Volunteer healthy blood donors can be a control population for assessment of SARS-CoV-2 exposure in the population. We determined COVID-19 seroprevalence during the second pandemic wave in Karachi in donors without known infections or symptoms in 4 weeks prior to enrollment. MATERIALS AND METHODS: We enrolled 558 healthy blood donors at the Aga Khan University Hospital between December 2020 and February 2021. ABO blood groups were determined. Serum IgG reactivity were measured to spike and receptor binding domain (RBD) proteins. RESULTS: Study subjects were predominantly males (99.1%) with a mean age of 29.0±7.4 years. Blood groups were represented by; B (35.8%), O (33.3%), A (23.8%) and AB (7%). Positive IgG responses to spike were detected in 53.4% (95% CI, 49.3-37.5) of blood donors. Positive IgG antibodies to RBD were present in 16.7% (95% CI; 13.6-19.8) of individuals. No significant difference was found between the frequency of IgG antibodies to spike or RBD across age groups. Frequencies of IgG to Spike and RBD antibodies between December 2020 and February 2021 were found to be similar. Seropositivity to either antigen between individuals of different blood groups did not differ. Notably, 31.2% of individuals with IgG antibodies to spike also had IgG antibodies to RBD. Amongst donors who had previously confirmed COVID-19 and were seropositive to spike, 40% had IgG to RBD. CONCLUSIONS: Our study provides insights into the seroprevalence of antibodies to COVID-19 in a healthy cohort in Karachi. The differential dynamics of IgG to spike and RBD likely represent both exposure to SARS-CoV-2 and associate with protective immunity in the population.

Combined central retinal artery and vein occlusion associated with antiphospholipid syndrome.

A female patient in early 50s presented to us several months after developing severe visual loss in her right eye. The patient was diagnosed with resolved central retinal vein occlusion (CRVO) based on the clinical picture at the time of presentation, however, retroactive evaluation of fundus imaging and further multi-disciplinary workup led to the rare diagnosis of combined central retinal artery and vein occlusion associated with antiphospholipid syndrome (APS). Only a few cases reporting retinal arterial and venous occlusions in patients with APS are found in the literature. To the best of our knowledge, no case of simultaneous CRAO and CRVO has been reported with APS. The patient was started on lifelong warfarin therapy to prevent a similar episode in the left eye. It is important to properly evaluate patients presenting with retinal vascular occlusions, as a missed diagnosis of APS can lead to recurrent and more devastating vascular events.

Perspective on newborn screening (NBS): Evidence sharing on conditions to be included in NBS in Pakistan.

Newborn screening aims at detecting treatable disorders early so that the treatment can be initiated to prevent mortality and morbidity. Such programmes are well established in most developed countries, and all newborns are screened for selected metabolic, endocrine and other disorders based on disease epidemiology, testing and treatment availability, efficiency and cost-effectiveness. Even in developing countries, such screening programmes are initiated using heel prick capillary blood collected on filter paper. The current narrative review was planned to provide a perspective with evidence in favour of starting newborn screening for different disorders. The programme project should be initiated nationwide, taking one disorder, congenital hypothyroidism, as the prototype and a newborn screening panel can then be extended to include other disorders. A task force should be set up to recommend disorders to be included in the panel, develop the national plan policies, and define procedures to strengthen the testing.

D-dimer Testing in COVID-19: From Basics to Clinical Application.

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Association of Factor V Leiden G1691A and Prothrombin gene G20210A mutations with adverse pregnancy outcomes.

OBJECTIVE: To determine the association of Factor V Leiden / prothrombin gene mutation in Pakistani women with adverse pregnancy outcomes. METHOD: The prospective study was conducted at the Aga Khan University Hospital, Karachi, from January 1 to December 31, 2016, and comprised females > 40 years having history of two or more foetal losses with no apparent aetiology. Restriction fragment length polymorphism- Polymerase chain reaction was performed using MnlI and HindIII restriction enzymes for factor V Leiden G1691A and prothrombin gene mutation G20210A. Females with two or more consecutive normal pregnancies were enrolled as the control group. Data was analysed using SPSS 19. RESULTS: Of the 172 participants with a mean age of 29.3±5.9 years (range: 19-38 years). 86(50%) each were healthy controls and those with recurrent pregnancy loss. There were 238 livebirths among the controls compared to 13 in the other group. Factor V Leiden G1691A was identified in 2(2.3%) women, and prothrombin gene mutation G20210A in 1(1.2%) woman in the patient group, while no mutation was identified in the control group. CONCLUSIONS: The prevalence of Factor V Leiden / prothrombin gene mutation in women with recurrent pregnancy loss was found to be very low.

Case Study of Pediatric Cerebral Sinus Venous Thrombosis Center of a Low Middle-Income Country.

Pediatric cerebral venous sinus thrombosis (CVST) is rare but a potentially fatal disease requiring its understanding in local setting. In this study, we observed the clinical course, management, and outcome of pediatric patients with sinus thrombosis in a tertiary care center at Pakistan. Patients between age 0 to 18 years of both genders diagnosed with sinus thrombosis during 2011 to 2020 were included. Data was collected through in-house computerized system and SPSS version 19 was used for analysis. Of 143492 pediatric admissions, 32 (21 males and 11 females) patients with a median (IQR) age of 4.5 years (0-16) had CVST. This is equivalent to 18.5 CVST events per million pediatric admissions. Adolescents were mostly affected, and the overall mortality was 7%. Primary underlying disorders were infections (59%), hematological neoplasms (12.5%), thrombotic thrombocytopenic purpura (3%) and antiphospholipid syndrome (3%). Activated protein C resistance (44%) was the most common inherited thrombophilia. Twenty-one (66%) patients were anemic with a mean (±SD) hemoglobin of 9.0 g/dL (±2.3). Regression analysis showed a positive association of anemia with multiple sinus involvement (P-value 0.009) but not with duration of symptoms (P-value 0.344), hospital stay (P-value 0.466), age (P-value 0.863) or gender (P-value 0.542) of the patients. SARS-COV2 was negative in patients during 2020. Adolescents were primarily affected by sinus thrombosis and infections was the predominant risk factor for all age groups, with a low all-cause mortality. A high index of clinical suspicion is required for prompt diagnosis and intervention.

Plasmodium in the bone marrow: case series from a hospital in Pakistan, 2007-2015.

BACKGROUND: Malaria is a life-threatening, multisystem disease caused by the plasmodial parasite with a global incidence of approximately 229 million annually. The parasites are known to have unique and crucial interactions with various body tissues during its life cycle, notably the liver, spleen, and recent work has shown the bone marrow to be a reservoir of infection. METHODS: This study is a case series of patients in whom examination of bone marrow revealed malarial parasites. A retrospective record review of 35 parasite-positive bone marrow specimens examined at Aga Khan University Hospital (AKUH), Karachi, Pakistan, over the years 2007 to 2015 was conducted. Bone marrow aspirates were collected as per International Council for Standardization in Haematology (ICSH) guidelines. RESULTS: The median age of patients was 22 years (range 1-75), and 60 % (n = 21) were male. 22 patients had evidence of Plasmodium falciparum, 12 had evidence of Plasmodium vivax and 1 patient had a mixed infection. Gametocytes and trophozoites were the most common stages identified on both peripheral blood and bone marrow examinations. Indications for bone marrow examination included fever of unknown origin and the workup of cytopenias and malignancies. CONCLUSIONS: The incidental finding of Plasmodium in samples of bone marrow suggests the reticuloendothelial system may be regularly harbour these parasites, be the infection acute or chronic in character.

Prolonged activated partial thromboplastin time secondary to factor XII deficiency in two surgical patients.

Factor XII (FXII) plays a pivotal role in hemostasis, inflammation and complement system. Its deficiency is usually an incidental finding in an otherwise asymptomatic patient who is identified during his/her routine preoperative blood work. This study aimed in evaluating the clinical course of the surgical patients having FXII deficiency. Information regarding demographics, laboratory tests and management of patients was obtained through medical chart and in-house integrated laboratory management system whereas the medical literature was searched through PubMed®. During the study period, two patients were consulted for FXII deficiency prior to the various surgical procedures. Both patients had uneventful surgeries without any thrombotic events while hemorrhage observed in one patient was secondary to obstetric complications. With the limited evidence today, it is concluded that patients having FXII deficiency are not at increased risk of bleeding, thrombosis or infections during surgery, but a personalized approach is needed for planning an appropriate perioperative management.

Risk Assessment and Outcome of Venous Thromboembolism in Pediatric Population in an Academic Care Center of a Low-Middle Income Country.

Venous thromboembolism (VTE) is a recognized complication of hospital stay in young patients in many developed countries, but such an information is largely unavailable from a low middle-income country (LMIC). This study aimed at identifying the frequency, risk factors, treatment options and outcome of deep venous thrombosis/pulmonary embolism (DVT/PE) in pediatric population in a tertiary care center from a LMIC. International classification of disease, ninth revision (ICD-9) was used to identify VTE in patients aged 0-18 years during January 2011 to September 2019. In-house computerized system was used to collect data for demographics, clinical and laboratory details. SPSS version 19 was used to analyzed data. The study was approved by Institutional ethical review committee (3872-Pat-ERC-15). During the study period, 134617 pediatric patients were hospitalized, DVT/PE was observed in 77 unique patients (47 males and 30 females) with a median (IQR) age of 14 (5-16) years equivalent to 5.9 VTE events /10,000 hospital admissions. Malignancy, community acquired infections and autoimmune diseases were the predominant risk factors (75%) in adolescent age-group while surgery for congenital heart anomalies was the primary reason (71%) in infants. Overall, lower extremity thrombosis was the most frequent (51%) followed by pulmonary embolism (25%). and upper extremity thrombosis (24%). Enoxaparin and unfractionated heparin were mainly used to treat VTE and all-cause mortality was 13% in the cohort studied. We observed substantial VTE events in pediatric patients during their hospital stay in a tertiary care center of a low-middle income country.

Documentation errors in transfusion chain: Challenges and interventions.

BACKGROUND AND OBJECTIVE: There are several steps in transfusion chain where accurate documentation is critical. This study was conducted to evaluate the frequency of documentation errors during transfusion process and to evaluate the effectiveness of interventions in error-management. METHODS /MATERIAL: This study was conducted at Aga Khan University, Pakistan during 2016-2018. Transcription and bedside documentation errors were identified from in-house computerized system and from medical charts. Raw WBIT rate was calculated for repeat blood samples and adjusted for frequencies of ABO-groups in our population accounting for silent WBIT. Rate of ABO-mismatched red cell transfusions was calculated for the annual totals of red cell transfusions. Chi-square was used for observing relationship among errors of various data sets. RESULTS: A total of 43 WBIT was identified during 54,219 repeat blood samples where blood group was already defined in blood bank information system. Annual unadjusted and cryptic WBIT rate was consistent at 0.8 and 0.6 per 1000 samples respectively during 2016-2018 (p 0.859). There were 1161 transcription errors (1.1 %) in blood group documentation in 105,064 blood samples received for arranging blood products. ABO-mismatched transfusion rate was 0.9 for 10,000 RBC transfusions in pre- and decreased to 0.4 in post-typing era. Overall, the compliance for completing checklist, correct ABO technique and appropriate ABO-interpretation was 88 %, 40 % and 24 % in the reviewed medical charts. CONCLUSIONS: Sample labeling errors were not improved through training or counseling. Bedside ABO-typing and checklist prior to blood transfusion can control the ABO-mismatched transfusion if done timely and correctly.

Reporting transfusion-related acute lung injury cases.

BACKGROUND: Transfusion-related acute lung injury (TRALI) is a rare but potentially fatal complication of blood product transfusion. It is felt worldwide that TRALI is an underrecognized and underreported entity because of lack of awareness. AIM: The purpose of this study was to report all cases of TRALI diagnosed in a tertiary care hospital over a 5-year period. MATERIALS AND METHODS: This is a retrospective review of all TRALI cases reported from January 2011 to December 2015. All TRALI cases were identified from a manual review of reported transfusion reaction forms. For detailed information of all TRALI cases, medical record charts of patients were reviewed. The record of donors implicated in TRALI cases was derived from blood bank system. STATISTICAL ANALYSIS USED: The rate of TRALI cases per 1000 blood products transfused was computed by dividing the transfusion reactions by total number of all blood units transfused. RESULTS: Total number of transfusions during the study was 291,041. Six cases of TRALI were reported during this period. Rate of TRALI per 1000 units transfused was 0.02%. The mortality associated with TRALI was 33.3%. TRALI occurred following the transfusion of fresh-frozen plasma in one patient, packed red blood cells in two patients, and a mixture of blood components in three patients. In all cases, the donors were male. CONCLUSION: The rate of TRALI reported to our blood bank was found to be 0.02%, which is very low as compared to international data. This is the first comprehensive study on TRALI from the country and a step forward to create awareness about the importance of diagnosing and reporting TRALI.

Development and Pilot Testing of a Novel Tool for Evaluating Practical Skills in Hematopathology Residents in Pakistan.

BACKGROUND: The mini-Clinical Evaluation Exercise (mini-CEX) and direct observation of procedural skills (DOPS) are reliable tools for work-based assessment of medical trainees. Tools of this type do not yet exist for evaluation of practical laboratory skills of pathology residents. OBJECTIVE: We developed and piloted a 9-item instrument for direct observation of laboratory skills (DOLS). METHODS: We used the DOLS tool with 10 hematopathology residents (PGY-1 to PGY-5) from Aga Khan University. Each resident was evaluated by 3 faculty members in the laboratory during 4 separate encounters using the DOLS instrument. We assessed construct validity, interrater reliability and G coefficient, feasibility of using DOLS, and learner satisfaction. RESULTS: A total of 120 encounters were observed with a mean score (±1 SD) of 56.7% (±12.44). Assessment scores moderately correlated with the number of laboratory procedures previously performed by participants (r = 0.658 and 0.641; P = .0001) and with PGY level. Interrater reliability ranged between 0.47 and 0.96. Cohen's d was 1.64. Residents accounted for a large component of estimated variance (73%), suggesting DOLS can differentiate residents' laboratory skills; variance associated with assessors was small (0.01%). Residents reported being satisfied with the tool. Mean time (±1 SD) taken for observing and feedback was 17.89 ± 5.89 minutes. CONCLUSIONS: The new DOLS instrument could provide reliable scores for observing laboratory skills. Residents were satisfied with the tool, and rating times make the tool feasible for formative assessments.

Unique classification of parathyroid dysfunction in patients with transfusion dependent thalassemia major using Nomogram-A cross sectional study.

INTRODUCTION: Hypoparathyroidism is a rare complication of iron overload in patients with transfusion dependent ß thalassemia major (ß-TM). We aim to determine the prevalence of parathyroid dysfunction in patients with ß-TM. METHODS: Diagnosed cases of transfusion dependent ß-TM between 5 and 17 years of age were recruited from outpatient clinics of a non-profit organization in Karachi, Pakistan. Blood and urine samples were collected in fasting to determine Ca, P, Alb, Mg, Cr 25OHD and iPTH. Patients were grouped on the basis of upper and lower levels of Ca, 25OHD and iPTH for assessing parathyroid dysfunction into primary hypoparathyroidism [low calcium (Ca) & intact parathyroid hormone (iPTH)], sub-clinical hypoparathyroidism [low iPTH and 25 hydroxy vitamin D (25OHD), low/normal Ca], normal functioning parathyroid gland [Normal Ca, iPTH and 25OHD] and secondary hyperparathyroidism [high iPTH, low/normal Ca and/or 25OHD]. Using PTH nomogram subject specific expected PTH (maxPTH) was calculated. Difference between maxPTH and measured iPTH was determined to assess the utility of nomogram in identifying parathyroid gland dysfunction. The statistical analysis was performed using the Statistical Package of Social Sciences (SPSS) version 20. RESULTS: Median age of patients was 11 years (13-7) with males being 54.2% (n = 205).Based on Ca, 25OHD and iPTH, primary hypoparathyroidism was identified in 3.4% (n = 13) [median iPTH 11.3 pg/ml (12.6-7)], 52.3% (n = 192) had subclinical hypoparathyroidism [iPTH 40.4 pg/ml (52.7-28.7)], and 34% (n = 125) were identified as secondary hyperparathyroidism [iPTH 88.6 pg/ml (116-74.7)]. Normal response to Ca & 25OHD was seen in 10.6% (n = 39) [iPTH 44.2 pg/ml (53.8-33.4)] patients. High phosphorous was present in all groups. Difference between maxPTH & iPTH was highest in primary hypoparathyroidism, followed by subclinical and secondary hyperparathyroidism. CONCLUSION: Nomogram by Harvey et al. identify low secretion capacity of parathyroid gland that correlated with biochemical classification of patients. It requires clinical validation before using in clinical practice for assessing parathyroid dysfunction.