A GP1BA Variant in a Czech Family with Monoallelic Bernard-Soulier Syndrome

Bernard-Soulier syndrome (BSS) is a rare inherited disorder characterized by unusually large platelets, low platelet count, and prolonged bleeding time. BSS is usually inherited in an autosomal recessive (AR) mode of inheritance due to a deficiency of the GPIb-IX-V complex also known as the von Willebrand factor (VWF) receptor. We investigated a family with macrothrombocytopenia, a mild bleeding tendency, slightly lowered platelet aggregation tests, and suspected autosomal dominant (AD) inheritance. We have detected a heterozygous GP1BA likely pathogenic variant, causing monoallelic BSS. A germline GP1BA gene variant (NM_000173:c.98G > A:p.C33Y), segregating with the macrothrombocytopenia, was detected by whole-exome sequencing. In silico analysis of the protein structure of the novel GPIbα variant revealed a potential structural defect, which could impact proper protein folding and subsequent binding to VWF. Flow cytometry, immunoblot, and electron microscopy demonstrated further differences between p.C33Y GP1BA carriers and healthy controls. Here, we provide a detailed insight into its clinical presentation and phenotype. Moreover, the here described case first presents an mBSS patient with two previous ischemic strokes.

Bismuth Subgallate as a Local Hemostatic Agent

Background: Patients on clopidogrel increased bleeding risk after surgery. This drug prolonged bleeding time, increased bleeding volume and induced secondary bleeding because its active metabolite inhibited platelets aggregation and interfered with haemostatic plug stabilization. Conventional methods, such as pressing sterile gauze on the surgery site, showed less effective to stop bleeding in patients on clopidogrel. This research aims to prove the haemostatic effect of bismuth subgallate both on normal and delayed platelet aggregation due to clopidogrel.Methods: Twenty-eight Wistar rats were equally and randomly administered with clopidogrel (10 mg/kgBW) or NaCl 0.9% (saline) via oral gavage. After anesthetizing, we amputated transversely their tail 10 mm from the distal tip. Bleeding after amputation was controlled with pressing gauze soaked in saline or bismuth subgallate solution. After 60 seconds, bleeding assays (bleeding time, bleeding volume, and secondary bleeding) have been observed, recorded, and analysed both in normal and clopidogrel groups.Results: Clopidogrel groups had significantly longer bleeding time, greater bleeding volume, and had more secondary bleeding rather than saline groups (p <.05). Using bismuth subgallate as local haemostatic agent decreased bleeding time and bleeding volume significantly (p <.05) both in normal and clopidogrel groups. Conclusions: Bismuth subgallate has a haemostatic effect on both clopidogrel and normal rat tail bleeding models.

A Rare Case of Bleeding Disorder: Glanzmann’s Thrombasthenia

Glanzmann’s thrombasthenia is an extremely rare autosomal recessive inherited bleeding disorder characterized by defective platelet aggregation leading to prolonged bleeding time. Patients may present with easy bruising, purpura, epistaxis, menorrhagia and gingival bleeding. Though the disease is rare, the prognosis is usually excellent with supportive care. Here, we report the case of Glanzmann’s thrombasthenia in a young female who presented with complaints of epistaxis and a history of easy bruising. The patient improved with symptomatic and supportive care. The patient got discharged and is doing well under regular follow-up.

EFFECTS OF HEMOTOXIC SNAKE BITE ENVENOMATION ON HAEMATOLOGICAL PARAMETERS VARIABILITY IN PREDICTING COMPLICATIONS

Background. Snake bite envenomation is a major public health problem in India with a high mortality rate. The major complications following a hemotoxic snake bite are disseminated intravascular coagulation (DIC), shock, acute kidney injury (AKI), acute respiratory distress syndrome (ARDS) and coagulopathy. The study explores a possible correlation of the haematological parameters studied to complications like DIC, AKI, acute renal failure (ARF), ARDS, shock and gastrointestinal (GI) bleed. Objective. The aim of the study was to find out the effect of snakebite envenomation on the major haematological parameters of the body in relation to complications. Methods. This cross-sectional study was conducted during a period of 18 months. 150 patients were included in the study after obtaining their informed consents. Data collection was done using a proforma. The study also compared clotting time (CT) by capillary tube method and whole blood clotting time at 20 minutes (WBCT20). SPSS software was used for statistical analysis. Results. Among the people with complications, the majority (52%) of participants had AKI, 26% of them requiring dialysis, 16.7% participants had GI bleed, 11.3% participants had shock and 10% participants had DIC. Conclusions. A prolonged bleeding time was found to be one of the most helpful haematological parameters in predicting shock and AKI. Clotting time by both capillary tube and WBCT20 methods were comparable in predicting complications. Objective: The objective of this study was to find the effect of snakebite envenomation on the major haematological parameters of the body. Material and method: This cross-sectional study was conducted over a period of 18months. A total of 150 patients were included in the study after obtaining informed consent. Data collection was done using a proforma. SPSS software was used for statistical analysis. Results: Among the people with complications, the majority (52%) of participants had AKI, followed by 26% participants who required Dialysis, 16.7% participants had GI bleed, 11.3% participants had the shock and 10% participants had DIC. Conclusion: A prolonged bleeding time was found to be one of the most helpful haematological parameters in predicting the shock and AKI.

Coagulation profile in cases of pre-eclampsia and eclampsia

Pregnancy Induced Hypertension (PIH) is a syndrome of hypertension with or without proteinuria (Preeclampsia) proteinuria and oedema. With additional symptoms like coagulation abnormalities and liver function abnormalities. Preeclampsia is a severe multi-systemic pregnancy-specific, hypertension disorder usually occurring after 20 weeks of gestation, characterised by new onset of hypertension and proteinuria, and it regresses after delivery of the concepts. The present cross-sectional case-control study was carried out in the Department of Pathology of a tertiary care hospital. Study of Coagulation profile in Preeclampsia and Eclampsia comprised of 258 cases which were categorised as preeclampsia, eclampsia and control group with 86 cases in each group. A total number of 258 patients were studied who were categorised as the control group, preeclampsia group and Eclampsia group with 86 cases in each. Preeclampsia was further classified as mild Preeclampsia (48/86) and severe Preeclampsia (38/86). Results-Majority of occurrences were noted in 21 to 25 years of age group, i.e. 45 patients of Eclampsia (52%), Eclampsia (67.44%) and Severe Preeclampsia (63.95%) were found to be more common in Primigravida patients. In Eclampsia group, 40% had mild thrombocytopenia. In Severe preeclampsia group, 37% had mild thrombocytopenia. In Mild preeclampsia group, 67% (32/48) cases showed platelet counts within a healthy range. In Eclampsia group, (13/86), i.e., 15% had prolonged bleeding time (>9 mints) In Severe preeclampsia group, (3/38), i.e. 8% had prolonged clotting time (>11mins) In Mild preeclampsia group and control group, none had prolonged clotting time (> 11mins). In Eclampsia group, (37/ 86), i.e., 43% had prolonged prothrombin time. Study concluded that coagulation profile could help to assess the severity of Preeclampsia and Eclampsia and thus can help to reduce complications if treated early.

Novel Mouse Model for Studying Hemostatic Function of Human Platelets

Objective: Platelets are critical to the formation of a hemostatic plug and the pathogenesis of atherothrombosis. Preclinical animal models, especially the mouse, provide an important platform to assess the efficacy and safety of antiplatelet drugs. However, these studies are limited by inherent differences between human and mouse platelets and the species-selectivity of many drugs. To circumvent these limitations, we developed a new protocol for the adoptive transfer of human platelets into thrombocytopenic nonobese diabetic/severe combined immune deficiency mice, that is, a model where all endogenous platelets are replaced by human platelets in mice accepting xenogeneic tissues. Approach and Results: To demonstrate the power of this new model, we visualized and quantified hemostatic plug formation and stability by intravital spinning disk confocal microscopy following laser ablation injury to the saphenous vein. Integrin α IIb β 3 -dependent hemostatic platelet plug formation was achieved within ≈30 seconds after laser ablation injury in humanized platelet mice. Pretreatment of mice with standard dual antiplatelet therapy (Aspirin+Ticagrelor) or PAR1 inhibitor, L-003959712 (an analog of vorapaxar), mildly prolonged the bleeding time and significantly reduced platelet adhesion to the site of injury. Consistent with findings from clinical trials, inhibition of PAR1 in combination with dual antiplatelet therapy markedly prolonged bleeding time in humanized platelet mice. Conclusions: We propose that this novel mouse model will provide a robust platform to test and predict the safety and efficacy of experimental antiplatelet drugs and to characterize the hemostatic function of synthetic, stored and patient platelets.

Novel Brown Coat Color (Cocoa) in French Bulldogs Results from a Nonsense Variant in HPS3

Brown or chocolate coat color in many mammalian species is frequently due to variants at the B locus or TYRP1 gene. In dogs, five different TYRP1 loss-of-function alleles have been described, which explain the vast majority of dogs with brown coat color. Recently, breeders and genetic testing laboratories identified brown French Bulldogs that did not carry any of the known mutant TYRP1 alleles. We sequenced the genome of a TYRP1+/+ brown French Bulldog and compared the data to 655 other canine genomes. A search for private variants revealed a nonsense variant in HPS3, c.2420G>A or p.(Trp807*). The brown dog was homozygous for the mutant allele at this variant. The HPS3 gene encodes a protein required for the correct biogenesis of lysosome-related organelles, including melanosomes. Variants in the human HPS3 gene cause Hermansky–Pudlak syndrome 3, which involves a mild form of oculocutaneous albinism and prolonged bleeding time. A variant in the murine Hps3 gene causes brown coat color in the cocoa mouse mutant. We genotyped a cohort of 373 French Bulldogs and found a strong association of the homozygous mutant HPS3 genotype with the brown coat color. The genotype–phenotype association and the comprehensive knowledge on HPS3 function from other species strongly suggests that HPS3:c.2420G>A is the causative variant for the observed brown coat color in French Bulldogs. In order to clearly distinguish HPS3-related from the TYRP1-related brown coat color, and in line with the murine nomenclature, we propose to designate this dog phenotype as “cocoa”, and the mutant allele as HPS3co.

Anti-Phospholipid Antibody Syndrome with Subdural Hematoma: A Case Report

A 33-year-old man with anti-phospholipid antibody syndrome associated with Budd Chiari syndrome and subdural hematoma. He developed venous thrombosis in his hepatic vein (stenting done) when laboratory studies demonstrated prolongation of activated partial thromboplastin time (APTT). Subdural hematoma demonstrated with Computed tomography (CT) of brain. Laboratory studies revealed thrombocytopenia, prolonged bleeding time and APTT, positive antinuclear antibody and positive test results for both lupus anticoagulant and an anti-cardiolipin antibody, namely antiphospholipid antibodies. Based on these findings, we consider that the tendency of this bleeding may have been due to antiphospholipid antibodies, attacking the platelet membranes and that the bridging veins in the subdural space may be the site at which the bleeding tendency easily appears. Antiphospholipid antibody syndrome accompanied by hemorrhagic complications had rarely been reported. We suggest that special attention should be given to hemorrhagic complications in patients with antiphospholipid antibody syndrome associated with fragility of the vessels and/or platelet dysfunction and on anticoagulant (warfarin).Pulse Vol.10 January-December 2017 p.25-28

ACETOSAL, BUAH MENGKUDU (MORINDA CITRIFOLIA L.) DAN WAKTU PERDARAHAN

Research regardless the effect of noni fruit for increasing bleeding time have already been carried out widely. The similar activity ofnoni fruit extract and acetosal can be concerned that the fruit extract has a potential activity for prolonged bleeding time. This study aimsto know the present of prolonged of bleeding time as a results of intake of combination of noni fruit extract with acetosal on mice. Thisresearch was carried out at Unit Binatang Percobaan, Departemen Farmakologi Universitas Udayana. This is an experimental study withpre and post-test control group design. Subject was compromised of 3 groups of mice and each group contain of 7 mice. The first groupwas treated with a dose of 40 mg/kg bw acetosal, the second group treated with a dose of 100 mg/kg bw ethanol noni fruit extract, andthe third group treated with combination of 40 mg/kg bw acetosal and 100 mg/kg bw ethanol extract of noni fruit. All groups werefed once per day for a week. Bleeding time was determined on the basis of tail bleeding method. This study results that the first groupexperience bleeding time increased from 61.42±9.43 second to 160.71±19.77 second. Increase bleeding time of the second group is from59.14±7.12 to 138.14±59.91 second. For the third group, the bleeding time increases from 65.00±7.91 to 213.00±20.92 second.One Way ANOVA analysis indicates that there is a significant different among these three groups after treatment p = 0.006 (p<0.05).Bleeding time of the third group which was treated with combination of noni fruit and acetosal results in the highest increase compareto the other two groups. In conclusions, combination of noni fruit and acetosal treatment results in increase of bleeding time on mice.