Thrombocytosis in children as well as in adult is defined as platelet count ≥ 450 × 109/L, and it is usually a reactive feature to various medical disorders. However, extreme thrombocytosis (platelet count ≥ 1000 × 109/L) is an uncommon finding among pediatric and adult patients, which may indicate more than a reactive phenomenon. We describe a case of a five-year-old boy who was admitted due to recurrent epistaxis. He had no history of allergic tendency or trauma. Physical examination was unremarkable except for shotty neck nodes. Laboratory results at presentation showed normal hemoglobin and total leukocyte count with eosinophilia (0.92 × 109/L), and extreme thrombocytosis. Other relevant investigations including coagulation profile, serum ferritin, liver, and renal function tests were all within normal ranges. Stool samples for ova and cysts were negative. The peripheral blood smear and bone marrow aspirate confirmed thrombocytosis with increased megakaryocytic proliferation and no artefactual reasons for the high platelets such as red blood cell fragments. Different causes of thrombocytosis in childhood were investigated after considering the possible differential diagnoses for extreme thrombocytosis.
The objective of this work is to characterize and investigate the blood compatibility of polyurethane (PU)/mustard oil composites fabricated using electrospinning technique. The fabricated scaffold was characterized using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), atomic force microscopy (AFM), thermogravimetric analysis (TGA) and contact angle measurements. The activated partial thromboplastin time (APPT), prothrombin time (PT) and the hemolytic assay were done to investigate the blood compatibility of the developed composites. The SEM results revealed that the fiber diameter of the composites (761 ± 123 nm) was reduced compared to pristine PU control. The interaction between PU and mustard oil was confirmed by FTIR as evident through the shifting of peaks. The fabricated composites depicted hydrophobic behavior as insinuated by the increase in contact angle measurements. PU/mustard composites displayed improved crystallinity as confirmed by TGA. Atomic force micrographs suggested that developed PU/mustard oil composites showed an increase in the surface roughness (Ra) compared to pure PU. The Ra of pure PU was observed to be 723 nm but for the fabricated PU/mustard oil composite the Ra was found to be 1298 nm (Ra). The hemolytic index value for pure PU and fabricated composites was observed to be 2.73% and 1.15% indicating that developed composites showed a non-hemolytic behavior signifying the safety of the composites with red blood cells. Hence the newly developed composites with improved physicochemical and blood compatibility properties may be considered as a potential candidate for fabricating cardiac patches and grafts.
To determine the optimum storage temperature and time for prothrombin time and activated partial thromboplastin time at various intervals at both room temperature and refrigerator.
The causes of an isolated prolonged activated partial thromboplastin time (APTT) with a normal prothrombin time (PT) are either a deficiency of clotting factors VIII, IX, XI or XII or the presence of an inhibitor. The inhibitor may be specific to an individual clotting factor or it may be a non-specific inhibitor like the lupus anticoagulant which has opposite therapeutic implications. We report a patient referred to our hospital for treatment that was previously diagnosed at another medical institution as an acquired factor IX inhibitor following an investigation for a prolonged APTT. On further testing this turned out to be a potent lupus anticoagulant which interfered with the phospholipid-dependent factor assays. The use of dilution studies, chromogenic assays and phospholipid neutralization can help differentiate these inhibitors. Great care must be taken in the interpretation of factor assays in the presence of lupus anticoagulant to avoid misdiagnosis and inappropriate treatment.
Bleeding following bites by the Malayan Pit Viper can either be local or systemic. Bleeding at the site of the bite is due to the local action of the venom as a vasculotoxin. Systemic bleeding occurs with severe poisoning and appears to be mainly dependent on platelet deficiency and the co-existing defibrination syndrome appears to play a minor role in the initiation of bleeding. Thus in the clinical situation non-clotting blood with no overt bleeding can continue up to weeks when specific antivenene is not given. Assessment of the severity of poisoning can easily be made at the bedside. Specific viper antivenene rapidly corrects the spontaneous bleeding and clotting defect of severe systemic poisoning but has no effect on local poisoning.
Congenital Coagulation Disorders (CCD) are inherited and present from birth. Their diagnosis depends on clinical awareness and correct laboratory investigations. The central registry for CCD or Congenital Bleeding Disorders (CBD) is at the Blood Services Centre, Kuala Lumpur Hospital and was established in 1975. There are 871 CCD registered. The commonest CCD are 631 (72%) Haemophilia A, 102 (12%) Haemophilia B and 93 (10.7%) von Willebrand's Disease. The other deficiencies registered are rare, only 45 in total:— Factor 1 (4), FV (4), FVII (21), FX (4), FXII (6), and FXIII (6). Diagnosis is based on clinical suspicion, screening tests namely the Prothrombin Time (PT) and activated Partial Thromboplastin Time (aPTT) and confirmation of the diagnosis was by doing specific factor assays. Molecular studies were done on FVIII and FXIII. Treatment is by transfusing the deficient factor when there is bleeding and comprehensive care involving the specialities like the neurologist/ neurosurgeon /orthopaedic / physiotherapy/ dental besides the haematologist and paediatrician to manage the complicatioons seen. There are fewer problems now as patients are diagnosed earlier and managed better. There is now a good prognosis and a better quality of life.
A well-accepted test for monitoring anticoagulation by enoxaparin is not currently available. As inadequate dosing may result in thrombosis or bleeding, a clinical need exists for a suitable test. Previous in silico and in vitro studies have identified factor Xa as an appropriate activating agent, and the phospholipid Actin FS as a cofactor for a Xa clotting time (TenaCT) test. A proof-of-concept study was designed to (1) explore the reproducibility of the TenaCT test and (2) explore factors that could affect the performance of the test. In vitro clotting time tests were carried out using plasma from 20 healthy volunteers. The effect of enoxaparin was determined at concentrations of 0.25, 0.50, and 1.0 IU/mL. Clotting times for the volunteers were significantly prolonged with increasing enoxaparin concentrations. Clotting times were significantly shortened for frozen plasma samples. No significant differences in prolongation of clotting times were observed between male and female volunteers or between the 2 evaluated age groups. The clotting times were consistent between 2 separate occasions. The TenaCT test was able to distinguish between the subtherapeutic and therapeutic concentrations of enoxaparin. Plasma should not be frozen prior to performing the test, without defining a frozen plasma reference range. This study provided proof-of-concept for a Xa-based test that can detect enoxaparin dose effects, but additional studies are needed to further develop the test.
Centrifugation of blood samples to produce platelet-poor plasma is one of the important steps for coagulation testing. Reduction of the time required for specimen processing without affecting quality of results should be ideal for tests which require immediate results. Centrifugation of platelet-poor plasma (3580 rpm) for 15 minutes performed for routine coagulation tests would prolong the turn-around time for an urgent test (30 minutes). This study was done to determine the effect of reducing centrifugation time for routine coagulation tests in order to meet the turn-around time (TAT) for urgent tests. Seventy-nine blood samples sent for routine coagulation tests, were assayed for prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen level and platelet counts, using two different centrifugation speed for plasma preparation: centrifugation at 3580 rpm for 15 minutes and rapid centrifugation at 4000 rpm for five minutes. Paired sample t-test showed that there was a significant
difference in the platelet count between the two groups (p=0.001). However, there was no significant difference in the normal APTT (p=0.16), abnormal APTT (p=0.80), abnormal PT (p=0.43) and the results of fibrinogen levels (p=0.36). In conclusion, rapid centrifugation at 4000 rpm for five minutes does not modify results of routine coagulation tests (PT, APTT and fibrinogen). It would be beneficial in providing rapid results for urgent coagulation tests.
Combined factor V and VIII deficiency is a rare bleeding disorder. Diagnosis of congenital coagulation factor deficiency in a neonate is challenging due to "immaturity" of the hemostatic system. A 2-day-old baby girl presented with spontaneous cephalhematoma. She was found to have persistent abnormal coagulation tests and finally diagnosed as combined factor V and VIII deficiency. Interestingly, factor V and factor VIII in developmental hemostasis are quite similar with adult levels in newborn, and hence early diagnosis is possible. An investigation to detect underlying hemostatic defects is recommended in newborns with spontaneous cephalhematoma.
Thawed plasma is fresh frozen plasma (FFP) that has been stored for 5 days at 1-6 °C. Duration of storage and different storage temperatures might affect the coagulation factor activity in thawed FFP. This study measured the changes of coagulation factor activities over 5 days in thawed FFP and stored at two different initial storage temperatures. Thirty-six units of FFP, which consisted of nine units each from blood groups A, B, AB, and O, were thawed at 37 °C. Each unit was divided into two separate groups (Group A and Group B) based on initial storage temperature. The first group was stored at 2-6 °C for 5 days (Group A). The second group was stored at 20-24 °C for initial 6 h followed by 2-6 °C for 5 days (Group B). Prothrombin time (PT), activated partial thromboplastin time (APTT), coagulation factor activities of fibrinogen, factor (F) II, FV, FVII, FVIII, FIX, FX, and von Willebrand factor antigen (vWF Ag) were assessed at baseline after thawing, at 6 h, and on days 1, 3, and 5 of storage for both groups. All coagulation factors mean activities in both storage groups decreased significantly over 5 days of storage. The mean FVIII activity at day 5 of storage was 36.9% in Group A and 39.8% in Group B. The other coagulation factors mean activities were > 50% on day 5 of storage in both groups. The coagulation factor activities of thawed FFP stored for 5 consecutive days were reduced in the two storage groups but most of the activities were still above 30%. This study suggests that thawed FFP stored for 5 days has the potential to ameliorate coagulation factor deficiencies in affected patients.
: Glanzmann's thrombasthenia is a rare inherited bleeding disorder characterized by the quantitative or qualitative defect of glycoprotein IIb/IIIa receptor on platelets which leads to ineffective aggregation. Light transmittance aggregometry is considered as the gold standard for diagnosis of Glanzmann's thrombasthenia. Thromboelastography (TEG) is a global hemostatic assay which measures clot formation, clot strengthening and fibrinolysis. This study evaluates the clinical, laboratory and TEG profiles in patients with Glanzmann's thrombasthenia. Bleeding score by (International Society on Thrombosis and Haemostasis) ISTH-bleeding assessment tool (bleeding score), laboratory tests to diagnose Glanzmann's thrombasthenia, and TEG parameters were correlated in 11 Glanzmann's thrombasthenia patients. Seventeen participants with normal bleeding score were included as controls. Bleeding score was increased in all patients. The highest bleeding score was in an adult female (26), whereas the lowest score (4) was in two children of less than 1 year. Majority of TEG parameters (except R-time) showed a statistically significant difference between Glanzmann's thrombasthenia patients and controls (K-time: P
Several bioactive molecules isolated from the saliva of blood-sucking arthropods, such as mosquitoes, have been shown to exhibit potential anticoagulant function. We have previously identified a 30kDa allergen named Aegyptin-like protein (alALP), which is highly homologous to Aegyptin, from the salivary glands of female Aedes albopictus (Asian tiger mosquito). In this study, we identified the conserved functional domain of alALP by using bioinformatic tools, and expressed the His-tagged alALP recombinant protein in sf9 insect cells by generation and transfection of a baculoviral expression plasmid carrying the fulllength cDNA of alALP. We purified this recombinant protein and examined its function on the inhibition of blood coagulation. The results showed that the purified His-alALP prolonged the Activated Partial Thromboplastin Time (APTT), Prothrombin Time (PT) and Thrombin Time (TT) in vitro as well as the Bleeding Time (BT) in vivo, which suggest that alALP could be a novel anticoagulant.
Green pit viper (Trimeresurus sp.) bite occurred throughout Myanmar, but there is no specific antivenom produced in the country for related envenomation. Instead, Myanmar Russell's viper antivenom (Anti-MRV) was often misused because of prolonged clotting time was observed from both species. Thai green pit viper antivenom (Anti-TGPV) raised against Trimeresurus albolabris was found to be effective against venoms of more than ten Trimeresurus sp. from Thailand, Malaysia and Indonesia. The present study compared the neutralization capacities of Anti-TGPV and Anti-MRV towards the venom from T. erythrurus from Myanmar. Anti-TGPV was more efficacious than Anti-MRV in cross-neutralizing the lethal and haemorrhagic activities of the venom by a potency of a least 1.4 times higher. Although Anti-TGPV effectively cross-neutralized the coagulation activity of the venom, Anti-MRV failed to do so. Immunodiffusion and immunoblot experiments showed that Anti-TGPV cross-reacted with more protein components of the venom than Anti-MRV. In conclusion, Anti-TGPV is a better choice for patients bitten by Myanmar green pit viper, but further clinical investigation is required. The current findings highlight the development of a specific antivenom against Myanmar green pit viper venom.
Conventional anticoagulant therapy is the mainstay of medical treatment for deep vein thrombosis disorders. However,there are many complications associated with these agents such as bleeding. Hence, the search for novel anticoagulant derived from natural substances such as plants origin is in high demand nowadays. Ocimum sanctum(O.sanctum) also known as Ocimum tenuiform (OT), tulsi or holy basil from the family of Lamiaceae has been widely used for thousands of years in Ayurveda and Unani systems to cure or prevent a number of illnessessuch as headache, malaria, ulcers, bronchitis, cough, flu, sore throat and asthma. The objective is to investigate theeffect ofO. sanctum(Tulsi) aqueous leaf extract on prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time (TT) in human plasma. Coagulation activity of O. sanctum was measured via PT, APTT and TT assay in citrated plasma collected from thirty-six healthy regular blood donors. The plasma was tested against different concentrations of O. sanctum aqueous extract as follows: 0.1mg/ml, 0.5 mg/ml and 1.0 mg/ml. Result shows the aqueous extract of O. sanctum prolonged the PT and APTT assays (p0.05). The gas chromatography-mass spectrometry (GC-MS) analysis had identified the linolenic acid at 1-10% of ethanol and aqueousconcentration at different retention time which was responsible for the coagulation activities of O. sanctumin human plasma. This study suggests that O. sanctum does affect coagulation activity in human plasma and can be potentially used as naturally derived anticoagulant products in the future.