Electronic medical record alert in patients with hemophilia at a teaching hospital emergency department

P157 Introduction: Patients with hemophilia are often not treated appropriately when they present out-of-hours to an emergency department (ED). Delays in replacement therapy can affect morbi- mortality. Clinical decision support systems include electronic medical record alerts (EMRA) and guidelines that assist in diagnosis and treatment. Aim: Audit the computerized automated alert tool in the electronic medical record in order to notify the need of health care provision of hemophilia patients in ED. Methods: Retrospective review (February 2015- March 2019) of health care alert notification emails regarding of hemophilia patients in our centre, received and generated by computer alert in the electronic medical record of the ED. Results: 164 visits to ED were registered, corresponding to 39 (37.5%) of the 104 hemophilia patients (pts) from our center. 28 of 80 pts with hemophilia A (HA) and 11 of 24 pts with HB. Median age 25 years (range: 5- 67). 31 were adults (median 33 years; range: 15-67) and 8 children under 15 (range 5- 14). Distribution by type and severity: HA: severe 9, moderate 2, mild 17; HB: severe 8, moderate 1, mild 2. 16 were under prophylaxis and 23 on demand. 11 went once to ED, 9 twice, 5 pts 3 times and 14 (36.6%) >5 times (5 with associated comorbidities). Causes for consultation (59 due to trauma and 57 directly related to hemophilia): Cutaneous/muscle bleed 18 episodes (Iliopsoas 5, cutaneous 5, other muscle 8), Joint pain 38 (bleed 8, synovitis 5, sprain 6, contusion 9, knee pain 2, post- traumatic fracture 3, nonspecific pain 5), abdominal pain 14 (gastroenteritis 5, gastrointestinal bleed 4, inguinal hernia 1, nonspecific pain 4), traumatic incise wound 4, infection 14, treatment administration 15, gingivorrhagia 7, colic pain/hematuria 7, toothache 4, allergic skin reaction 4, epistaxis 3, head trauma 2 and other causes not related to hemophilia 34 episodes. 23 hospital admissions were made in16 pts. The average length of stay of all patients admitted to ED was 4.8 hours as opposed to 2.4 hours (range: 0.1- 44.3) in hemophiliacs. Discussion/Conclusion: In our experience, only one third of patients consult with symptoms related to hemophilia at ED. The reason for hospital admission is mostly associated to other comorbidities. EMRA system allows early care provision, better compliance with the healthcare protocol, and shorten the length of stay and reducing morbidity in hemophilia patients

Unraveling the effect of silent, intronic and missense mutations on VWF splicing: contribution of next generation sequencing in the study of mRNA

Large studies in von Willebrand disease patients, including Spanish and Portuguese registries, led to identification of >250 different mutations. It is a challenge to determine the pathogenic effect of potential splice site mutations on VWF mRNA. This study aimed to elucidate the true effects of 18 mutations on VWF mRNA processing, investigate the contribution of next-generation sequencing to in vivo mRNA study in von Willebrand disease, and compare the findings with in silico prediction. RNA extracted from patient platelets and leukocytes was amplified by RT-PCR and sequenced using Sanger and next generation sequencing techniques. Eight mutations affected VWF splicing: c.1533+1G>A, c.5664+2T>C and c.546G>A (p.=) prompted exon skipping; c.3223-7_3236dup and c.7082-2A>G resulted in activation of cryptic sites; c.3379+1G>A and c.7473G>A (p.=) demonstrated both molecular pathogenic mechanisms simultaneously; and the p.Cys370Tyr missense mutation generated two aberrant transcripts. Of note, the complete effect of 3 mutations was provided by next generation sequencing alone because of low expression of the aberrant transcripts. In the remaining 10 mutations, no effect was elucidated in the experiments. However, the differential findings obtained in platelets and leukocytes provided substantial evidence that 4 of these would have an effect on VWF levels. In this first report using next generation sequencing technology to unravel the effects of VWF mutations on splicing, the technique yielded valuable information. Our data bring to light the importance of studying the effect of synonymous and missense mutations on VWF splicing to improve the current knowledge of the molecular mechanisms behind von Willebrand disease.info:eu-repo/semantics/publishedVersio

Unraveling the effect of silent, intronic and missense mutations on VWF splicing: contribution of next generation sequencing in the study of mRNA

Large studies in von Willebrand disease patients, including Spanish and Portuguese registries, led to the identification of >250 different mutations. It is a challenge to determine the pathogenic effect of potential splice site mutations on VWF mRNA. This study aimed to elucidate the true effects of 18 mutations on VWF mRNA processing, investigate the contribution of next-generation sequencing to in vivo mRNA study in von Willebrand disease, and compare the findings with in silico prediction. RNA extracted from patient platelets and leukocytes was amplified by RT-PCR and sequenced using Sanger and next generation sequencing techniques. Eight mutations affected VWF splicing: c.1533+1G>A, c.5664+2T>C and c.546G>A (p.=) prompted exon skipping; c.3223-7_3236dup and c.7082-2A>G resulted in activation of cryptic sites; c.3379+1G>A and c.7437G>A) demonstrated both molecular pathogenic mechanisms simultaneously; and the p.Cys370Tyr missense mutation generated two aberrant transcripts. Of note, the complete effect of three mutations was provided by next generation sequencing alone because of low expression of the aberrant transcripts. In the remaining 10 mutations, no effect was elucidated in the experiments. However, the differential findings obtained in platelets and leukocytes provided substantial evidence that four of these would have an effect on VWF levels. In this first report using next generation sequencing technology to unravel the effects of VWF mutations on splicing, the technique yielded valuable information. Our data bring to light the importance of studying the effect of synonymous and missense mutations on VWF splicing to improve the current knowledge of the molecular mechanisms behind von Willebrand disease. clinicaltrials.gov identifier:02869074