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.

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.

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Current COVID-19 pandemic has affected the entire globe. While there was no vaccine neither any specific treatment, investigational use of convalescent plasma has been explored in clinical trials. A prospective multicenter study of convalescent plasma was conducted. Donors were tested for total Anti-SARS-CoV-2 antibodies by electrochemiluminescence (ECLIA) and RT-PCR for COVID-19. Enzyme Linked Immunosorbent Assay (ELISA) was used to detect semi-quantitative and quantitative IgG anti-SARS-COV-2 antibodies. IgG Immunofluorescence-based lateral flow immunoassay (LFIA) was used to recheck seronegative donors. A total of 400 donors were enrolled. Twelve donors were SARS-CoV-2 positive by RT-PCR. Nine of 12 donors had developed SARS-CoV-2 IgG antibodies, while in 3 donors antibodies were not developed. A total of 70 donors (17.5%) were deferred due to seronegative status; 64 (16%) of them did not develop antibodies when plasma collection was planned. The IgG semiquantitative ELISA was positive in 282 and quantitative in 284 of 330 donors with a mean value of >1:160 and 44.10±39.22 IU/ml respectively. A total of 116 (29%) donors did not show IgG humoral response to COVID-19 even 28 days from the onset of illness. Subsequently, LFIA method was able to detect IgG antibodies in 20 of 48 (41.6%) seronegative donors and in 20 of 34 (58.8%) ECLIA positive ELISA negative donors. Viral RNA detection in recovered asymptomatic patients with concomitant IgG antibodies indicates recovery. Inability to detect antibodies by different testing kits may be due to their different antigenic targets or sensitivity. Significance of a positive COVID-19 RT-PCR in asymptomatic recovered patients is yet to be determined.