BackgroundCongenital 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.MethodsThis 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.ResultsTen 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.ConclusionsCongenital 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.

Background Convalescent plasma(CP) was utilized as potential therapy during COVID-19 pandemic in Pakistan. The study aimed at appraisal of CP transfusion safety and usefulness in COVID pneumonia. Methods Single arm, MEURI study design of non-randomized open label trial was conducted in five centers. Patients werecategorized as moderately severe, severe, and critical. The primary endpoint was a) improvement in clinical status and change in category of disease severity; secondary endpoint was b) CP ability to halt disease progression to invasive ventilation. CP transfused to hospitalized patients. Statistical tests including median (interquartile ranges), Mann-Whitney U test, Fisher’s exact test using SPSS ver. 23, ANOVA and Chi-square test were applied for the analysis of results parameters before and after CP treatment. SOFA score was applied for multiorgan failure in severe and critical cases. Results A total of 50 adult patients; median age 58.5 years (range: 29–92 years) received CP with infusion titers; median 1:320 U/mL (Interquartile range 1:80–1:320) between April 4 to May 5, 2020. The median time from onset of symptoms to enrollment in trial was 3 to 7 days with shortness of breath and lung infiltration as severity criterion. In 35 (70%) recipients, oxygen saturation improved from 80 to 95% within 72h, with resolution of lung infiltrates. Primary endpoint was achieved in 44 (88%) recipients whereas secondary endpoint was achieved in 42 (84%). No patient experienced severe adverse events. A high SOFA score (> 7) correlated with deaths in severe and critical patients. Eight (16%) patients expired due to comorbidities; cardiac arrest in 2 (4%), multiorgan failure secondary to cytokine storm in 5 (10%) and ventilator associated complications in 1 (2%). Conclusion CP transfusion can be used as a safe and useful treatment in moderately severe and severe patients. Trial registration The trial registration number is NCT04352751 (https://www.irct.ir/search/result?query=IRCT20200414047072N1). Trial Registration date is 28th April 2020.

Diabetes Mellitus (DM) is the most frequently occurring metabolic disorder, caused by inadequacy in secretion of insulin or malfunction leading to chronic hyperglycemia.  Well-established corroborations have been reported in the literature suggesting association of ABO blood group with DM. The available literature focuses on adult population, with limited information of said association in children. Thus this study was aimed to determine the association of ABO blood group with Diabetes in pediatric patients having confirmed diagnosis. This was a comparative cross sectional study conducted from October 2021-January 2022 at Endocrinology Ward, Children Hospital Lahore (CHL), Pakistan. The study was approved by the local research ethics committee of CHL and conducted according to the declaration of Helsinki 2000. Blood grouping was done by forward and reverse methods. A total of 25 patients, including 18 males and 7 females were included. Frequency of blood group B (n=10;40%) was highest followed by blood group O (n=4;16%), A blood group (n=3;12%), and AB (n=1;4%) in males. In females, the blood group AB (n=3;12%) has the highest frequency followed by O (n=2;8%). Blood group A (n=1;4%) and B (n=1;4%) had the  same frequency among female diabetic patients

This study was aimed to compare hemostatic, fibrinolytic and thrombotic parameters in pre and post induction chemotherapy in acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL). A total of 110 diagnosed acute leukemia patients and 40 normal individuals were enrolled in the study. Questionnaire were filled and patients’ blood specimens were collected before the commencement and post induction chemotherapy. Prothrombin time (PT), activated partial thromboplastin time (aPTT), von Willebrand factor antigen (vWF Ag), fibrinogen level, factor VIIIc (FVIIIc), D-Dimers, fibrinogen degradation products (FDPs), anti-thrombin III (AT), lupus anticoagulant (LA), protein C (PC), protein S and diluted russel viper venom test (DRVVT) were conducted in all the participants. A significant rise was identified in post-induction levels of PT, aPTT, vWF Ag, fibrinogen and factor VIII in acute leukemia patients. Analysis of fibrinolytic markers indicated increased D-Dimers and plasminogen levels while the levels of alpha 2 anti-plasmin were reduced in pretreated patients. Thrombotic markers’ assessment showed increased levels of AT and LA while decreased level of PC in pretreated acute leukemia patients. It was concluded that remission induction chemotherapy, in acute leukemia patients, significantly affects the coagulation, fibrinolytic and thrombotic parameters.

Background: Glanzmann thrombasthenia (GT) is most common inherited platelet functional defect. It is an autosomal recessive disorder, characterized by a bleeding diathesis. Incidence is increased in locations where consanguineous marriages are common. The defect is caused by mutations in the genes encoding ITGA2B or ITGB3. This results in qualitative or quantitative abnormalities of the platelet receptor, αIIb-β3 integrin. Objectives: The aim of this study was to identify and correlate the mutations in GT patients with phenotype of the patient. Subjects and methods: 20 patients with GT were enrolled in the study to identify the molecular defects and to correlate their phenotype with their genotype. CBC with peripheral film, PT, APTT and Fibrinogen levels were done initially. Platelet aggregation studies, flow cytometry, and mutation analysis was done by Sanger sequencing. Genomic DNA was extracted from peripheral blood by QIAamp DNA Blood mini kit (Qiagen) and Exon specific PCR was done for GT gene and Direct gene sequenced on automated ABI-3130 Genetic Analyzer (Applied Biosystems). For any variation wild type was matched on HGMD (Human Gene Mutation Database http://www.hgmd.cf.ac.uk/ac/index.php) and wild type color fasta sequence (http://pga.gs.washington.edu/). Pathogenecity score was evaluated by using software tools including : Polyphen-2(http://genetics.bwh.harvard.edu/pph2/) ,SNP&GO(http://snps.biofold.org/snps-and-go/index.html), MUpro (http://mupro.proteomics.ics.uci.edu/) ,SIFT (http://sift.jcvi.org), Provean (http://provean.jcvi.org/about.php) . All samples were sequenced at the Gene Sequencing Lab of NIBD (National Institute of Blood diseases and Bone Marrow Transplantation) Karachi. Results: Mutations were identified in all patients. Missense mutations were seen in most of the GT patients. The remaining mutations were heterogeneous and were distributed throughout the length of the gene. Conclusions: The severe type I GT was the most common subtype found in this study.Carrier detection and genetic counseling in these families is a potentially effective alternative for decreasing the burden of severe type of GT. Disclosures No relevant conflicts of interest to declare.