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The role of peripheral blood smear examination in the evaluation of suspected platelet-related disorders in children: A practical approach and an illustrated review

Tyrrell L, Scruggs M, Kerwin A, Kahwash SB

Malays J Pathol, 2022 Dec;44(3):397-413.
PMID: 36591709

Platelets, along with coagulation factors and vasculature, represent the three main compartments of hemostasis. Upon investigation of a suspected hemostasis disorder, platelet count, size and morphology often offer important clues to the diagnosis or help narrow the differential diagnosis. In this review, we describe a general approach to diagnosing platelet disorders, starting with easily obtained data such as findings of complete blood count (CBC) and microscopic review of a stained peripheral blood smear. We discuss general findings that help separate consumptive from underproduction thrombocytopenia. We further touch on inherited thrombocytopenia disorders after classifying them into those associated with small, normal sized or large platelets. Illustrative microscopic images are provided where contributory. We conclude with a suggested algorithmic step-by-step approach to investigating a suspected platelet disorder in children.

Matched MeSH terms: Blood Coagulation Disorders*
Does automation play a role in coagulation studies in Southeast Asian countries

George E

Southeast Asian J. Trop. Med. Public Health, 1979 Sep;10(3):319-20.
PMID: 515794
Matched MeSH terms: Blood Coagulation Disorders/diagnosis; Blood Coagulation Tests/instrumentation; Blood Coagulation Tests/methods*
Blood coagulation, fibrinolysis and coronary heart disease: observations in Malaysia

Loncin H, Gurian JM, Loncin ME

J Atheroscler Res, 1968 5 1;8(3):471-82.
PMID: 5660508
Matched MeSH terms: Blood Coagulation*; Blood Coagulation Disorders/epidemiology*; Blood Coagulation Tests
Biosensing human blood clotting factor by dual probes: Evaluation by deep long short-term memory networks in time series forecasting

Gopinath SCB, Ismail ZH, Shapiai MI, Sobran NMM

Biotechnol Appl Biochem, 2021 Apr 09.
PMID: 33835514 DOI: 10.1002/bab.2164

Artificial intelligence of things (AIoT) has become a potential tool for use in a wide range of fields, and its use is expanding in interdisciplinary sciences. On the other hand, in a clinical scenario, human blood-clotting disease (Royal disease) detection has been considered an urgent issue that has to be solved. This study uses AIoT with deep long short-term memory networks for biosensing application and analyzes the potent clinical target, human blood clotting factor IX, by its aptamer/antibody as the probe on the microscaled fingers and gaps of the interdigitated electrode. The earlier results by the current-volt measurements have shown the changes in the surface modification. The limit of detection (LOD) was noticed as 1 pM with the antibody as the probe, whereas the aptamer behaved better with the LOD at 100 fM. The time-series predictions from the AIoT application supported the obtained results with the laboratory analyses using both probes. This application clearly supports the results obtained from the interdigitated electrode sensor as aptamer to be the better option for analyzing the blood clotting defects. The current study supports a great implementation of AIoT in sensing application and can be followed for other clinical biomarkers.

Matched MeSH terms: Blood Coagulation; Blood Coagulation Disorders; Blood Coagulation Factors
Aptamer-antibody dual probes on single-walled carbon nanotube bridged dielectrode: Comparative analysis on human blood clotting factor

Yao J, Li S, Zhang L, Yang Y, Gopinath SCB, Lakshmipriya T, et al.

Int J Biol Macromol, 2020 May 15;151:1133-1138.
PMID: 31743722 DOI: 10.1016/j.ijbiomac.2019.10.156

Haemophilia is a blood clotting disorder known as 'Christmas disease' caused when the blood has defect with the clotting factor(s). Bleeding leads various issues, such as chronic pain, arthritis and a serious complication during the surgery. Identifying this disease is mandatory to take the necessary treatment and maintains the normal clotting. It has been proved that the level of factor IX (FIX) is lesser with haemophilia patient and the attempt here is focused to quantify FIX level by interdigitated electrode (IDE) sensor. Single-walled carbon nanotube (SWCNT) was utilized to modify IDE sensing surface. On this surface, dual probing was evaluated with aptamer and antibody to bring the possible advantages. The detection limit with antibody was found to be 1 pM, while aptamer shows 100 fM. Further, a fine-tuning was attempted with sandwich pattern of aptamer-FIX-antibody and antibody-FIX-aptamer and compared. Specific elevation of detection with 10 folds was noticed and displayed the detection at 100 f. in both sandwich patterns. In addition, FIX was detected in the diluted human serum by aptamer-FIX-antibody sandwich, it was found that FIX detected from the dilution factor 1:640. A novel demonstration is with higher discrimination against other clotting factors, XI and VII.

Matched MeSH terms: Blood Coagulation Factors; Blood Coagulation Tests*
Potential blood clotting factors and anticoagulants

Jin NZ, Gopinath SCB

Biomed Pharmacother, 2016 Dec;84:356-365.
PMID: 27668535 DOI: 10.1016/j.biopha.2016.09.057

Hemostasis initiates a wound healing process and stops bleeding of blood within a damaged tissue, an important process in human and animal systems. However, this process needs to revert temporarily during surgery and analyze the clotting mechanism. In the past decade, heparin has been used widely as an anticoagulant in surgery to prevent unwanted blood clotting as it is not expensive, not difficult to control, lack of suitable replacement as well as less harmful to the human. However, heparin has several disadvantages, which include thrombocytopenia and non-specific plasma binding. Moreover, using heparin it may lead dysfunction and platelet aggregation. In this overview, potential clotting factors and anticoagulants are reviewed and special focus was given to get more insights.

Matched MeSH terms: Blood Coagulation; Blood Coagulation Factors/metabolism*
Anticoagulant activity of heat-unmasked antiphospholipid autoantibody in normal human plasma?

Ismail R, Cheng HM

Thromb Res, 1994 Jan 15;73(2):143-7.
PMID: 8171414 DOI: 10.1016/0049-3848(94)90090-6
Matched MeSH terms: Blood Coagulation/drug effects*
Fulltext Effect of higher centrifugation speed and shortened centrifugation time on prothrombin and activated thromboplastin time

Azlin, I., Hafiza, A., Azma, R.Z., Aidifitrina, R.K., Hamidah, N.H.

Medicine & Health, 2011;6(1):68-72.
MyJurnal

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.

Matched MeSH terms: Blood Coagulation; Blood Coagulation Tests
HAEMOPHILIA DUE TO FACTOR 8 DEFICIENCY IN AN INDIAN BOY

ENG LI, LONCIN M, PILLAY RP

Med J Malaysia, 1964 Jun;18:219-22.
PMID: 14199436
Matched MeSH terms: Blood Coagulation Tests*
LABORATORY CONTROL OF ANTICOAGULANT THERAPY

LAU KS, SINGH N

Med J Malaysia, 1963 Dec;18:107-15.
PMID: 14117278
Matched MeSH terms: Blood Coagulation Tests*
A case of fatal envenomation by a captive puff adder (Bitis arietans) in Malaysia

Husain Z, Wicaksono AC, Renault A, Md Zhahir SS, Ismail AK

Toxicon, 2023 Mar 01;224:107023.
PMID: 36640813 DOI: 10.1016/j.toxicon.2023.107023

The Puff Adder (Bitis arietans) is a viper native to Africa and the Middle East. Envenomation by this species often requires the administration of appropriate antivenom in order to achieve a favorable outcome. A patient was bitten in both hands by a captive B. arietans presented to a teaching hospital in Malaysia. The patient developed painful progressive swelling on both limbs that extended to the chest, hypotension, hypokalemia with worsening anemia, thrombocytopenia, coagulopathy, and severe metabolic acidosis. The patient was managed supportively while waiting for the appropriate antivenom, Antivipmyn-Africa, from the Singapore Zoo. The patient developed cardiorespiratory arrest twice and did not recover from the second. The patient was pronounced dead 23 hours post-incident. The local unavailability of the appropriate antivenom may be the most important factor that contributed to the patient's death. There is also a need to amend the Malaysian Wildlife Act in order to prevent such cases from recurring.

Matched MeSH terms: Blood Coagulation Disorders*
Dental management of people with complex or rare inherited bleeding disorders

Nanayakkara L, Yahaya N, Parreira M, Bajkin B

Haemophilia, 2024 Apr;30 Suppl 3:128-134.
PMID: 38571337 DOI: 10.1111/hae.15005

Advances in haematological therapies for people with complex or rare inherited bleeding disorders (IBD) have resulted in them living longer, retaining their natural teeth with greater expectations of function and aesthetics. Dental management strategies need to evolve to meet these challenges. Utilising low level laser diode therapy to reduce pre-operative inflammation to reduce the intraoperative and postoperative burden on haemostasis is described in a case series of 12 patients. For these individuals who previously required further medical management to support haemostasis or experienced such prolonged haemorrhage sufficient to warrant hospital admission, haemostasis was achieved in the dental surgery such that they were able to return home with no further medical intervention or overnight stays. Global inequities in accessing novel treatments for complex or rare IBD necessitates a comprehensive understanding of the local haemostatic agents available to dentists and the most commonly used agents and techniques are described including the use of single tooth anaesthesia (STA). STA is a computerised delivery mechanism that allows routine dental procedures that would previously have required block injections needing factor replacement therapy to be undertaken safely and effectively with no additional haemostatic intervention. The challenges of inhibitors in oral surgery are explained and discussed although more research and evidence is required to establish new treatment protocols. The importance of establishing good dental health in the quality of life of people with complex or rare IBD is highlighted with respect to the dental specific impact that more novel therapies may have on people with IBD.

Matched MeSH terms: Blood Coagulation Disorders, Inherited*
Desmopressin acetate (DDAVP) for preventing and treating acute bleeds during pregnancy in women with congenital bleeding disorders

Karanth L, Barua A, Kanagasabai S, Nair S

Cochrane Database Syst Rev, 2015 Sep 09.
PMID: 26350784 DOI: 10.1002/14651858.CD009824.pub3

BACKGROUND: Congenital bleeding disorders can cause obstetric haemorrhage during pregnancy, labour and following delivery. Desmopressin acetate is found to be an effective drug which can reduce the risk of haemorrhage and can also stop bleeding in certain congenital bleeding disorders. Its use in pregnancy has been controversial. Hence beneficial and adverse effects of desmopressin acetate in these groups of pregnant women should be evaluated.This is an update of a Cochrane review first published in 2013.
OBJECTIVES: To determine the efficacy of desmopressin acetate in preventing and treating acute bleeds during pregnancy in women with congenital bleeding disorders.
SEARCH METHODS: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group's Coaguopathies Trials Register comprising references identified from comprehensive electronic database searches and handsearches of relevant and abstract books of conferences proceedings. We also searched for any randomised controlled trials in a registry of ongoing trials and the reference lists of relevant articles and reviews.Date of most recent search: 18 June 2015.
SELECTION CRITERIA: Randomised and quasi-randomised controlled trials investigating the efficacy of desmopressin acetate versus tranexamic acid or factor VIII or rFactor VII or fresh frozen plasma in preventing and treating congenital bleeding disorders during pregnancy were eligible.
DATA COLLECTION AND ANALYSIS: No trials matching the selection criteria were eligible for inclusion.
MAIN RESULTS: No trials matching the selection criteria were eligible for inclusion.
AUTHORS' CONCLUSIONS: The review did not identify any randomised controlled trials investigating the relative effectiveness of desmopressin acetate for bleeding during pregnancy in women with congenital bleeding disorders. In the absence of high quality evidence, clinicians need to use their clinical judgement and lower level evidence (e.g. from observational trials) to decide whether or not to treat women with congenital bleeding disorders with desmopressin acetate.Given the ethical considerations, future randomised controlled trials are unlikely. However, other high quality controlled studies (such as risk allocation designs, sequential design, parallel cohort design) to investigate the risks and benefits of using desmopressin acetate in this population are needed.

Matched MeSH terms: Blood Coagulation Disorders/congenital; Blood Coagulation Disorders/drug therapy*; Blood Coagulation Disorders/prevention & control*
An Evaluation of a Factor Xa-Based Clotting Time Test for Enoxaparin: A Proof-of-Concept Study

Ng DPJ, Duffull SB, Faed JM, Isbister GK, Gulati A

Clin Appl Thromb Hemost, 2018 May;24(4):669-676.
PMID: 28731370 DOI: 10.1177/1076029617711802

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.

Matched MeSH terms: Blood Coagulation/drug effects*; Blood Coagulation Tests/methods*
Fulltext Desmopressin acetate (DDAVP) for preventing and treating acute bleeds during pregnancy in women with congenital bleeding disorders

Karanth L, Barua A, Kanagasabai S, Nair NS

Cochrane Database Syst Rev, 2019 02 13;2:CD009824.
PMID: 30758840 DOI: 10.1002/14651858.CD009824.pub4

BACKGROUND: Congenital bleeding disorders can cause obstetric haemorrhage during pregnancy, labour and following delivery. Desmopressin acetate (DDAVP) is found to be an effective drug which can reduce the risk of haemorrhage and can also stop bleeding in certain congenital bleeding disorders. Its use in pregnancy has been controversial. Hence beneficial and adverse effects of DDAVP in these groups of pregnant women should be evaluated.This is an update of a Cochrane Review first published in 2013 and updated in 2015.
OBJECTIVES: To evaluate the efficacy and safety of DDAVP in preventing and treating acute bleeding in pregnant women with bleeding disorders.
SEARCH METHODS: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group's Coaguopathies Trials Register comprising references identified from comprehensive electronic database searches and handsearches of relevant and abstract books of conferences proceedings. We also searched several clinical trial registries and grey literature (27 August 2017).Date of most recent search of the Cochrane Cystic Fibrosis and Genetic Disorders Group's Coaguopathies Trials Register: 01 October 2018.
SELECTION CRITERIA: Randomised and quasi-randomised controlled trials investigating the efficacy of DDAVP versus tranexamic acid or factor VIII or rFactor VII or fresh frozen plasma in preventing and treating congenital bleeding disorders during pregnancy were eligible.
DATA COLLECTION AND ANALYSIS: No trials matching the selection criteria were eligible for inclusion.
MAIN RESULTS: No trials matching the selection criteria were eligible for inclusion.
AUTHORS' CONCLUSIONS: No randomised controlled trials were identified investigating the relative effectiveness of DDAVP for bleeding during pregnancy in women with congenital bleeding disorders. In the absence of high-quality evidence, clinicians need to use their clinical judgement and lower level evidence (e.g. from observational trials) to decide whether or not to treat women with congenital bleeding disorders with DDAVP.Given the ethical considerations, future randomised controlled trials are unlikely. However, other high-quality controlled studies (such as risk allocation designs, sequential design, parallel cohort design) to investigate the risks and benefits of using DDAVP in this population are needed.Given that there are unlikely to be any trials published in this area, this review will no longer be regularly updated.

Matched MeSH terms: Blood Coagulation Disorders/congenital; Blood Coagulation Disorders/drug therapy*; Blood Coagulation Disorders/prevention & control*
A potent anticoagulant hybrid of snake venom derived FIX-binding protein and anti-factor IX RNA aptamer: Assessed by in-silico and electrochemical analyses

Krishnan H, Gopinath SCB

Int J Biol Macromol, 2023 Aug 30;247:125740.
PMID: 37423441 DOI: 10.1016/j.ijbiomac.2023.125740

Anticoagulant therapies are crucial in the management of surgical complications as well as the prophylaxis of thrombosis. Many studies are being conducted on the Habu snake-venom anticoagulant, FIX-binding protein (FIX-Bp), for its greater potency and strong affinity to FIX clotting factor. On the other hand, the capacity to promptly reverse such acute anticoagulation is equally important. Combining a reversible anticoagulant with FIX-Bp may be advantageous in maintaining the balance between adequate anticoagulation and repealing when necessary. In this study, authors integrated FIX-Bp and RNA aptamer-based anticoagulants into a single target, FIX clotting factor, in order to achieve a robust anticoagulant effect. An in-silico and electrochemical approach were used to investigate the combination of FIX-Bp and RNA aptamers as a bivalent anticoagulant and to verify the competing or predominant binding sites of each anticoagulant. The in-silico analysis discovered that both the venom- and aptamer-anticoagulant had a strong affinity for the FIX protein at the Gla-domain and EGF-1 domain by holding 9 conventional hydrogen bonds with the binding energy of -34.859 kcal/mol. The electrochemical technique verified that both anticoagulants had different binding sites. The impedance load upon RNA aptamer binding to FIX protein was 14 %, whereas the addition of FIX-Bp caused a significant impedance rise of 37 %. This indicates that the addition of aptamers prior to FIX-Bp is a promising strategy for the conception of a hybrid anticoagulant.

Matched MeSH terms: Blood Coagulation Factors
Immune-stealth VP28-conjugated heparin nanoparticles for enhanced and reversible anticoagulation

Hussein HR, Chang CY, Zheng Y, Yang CY, Li LH, Lee YT, et al.

Nanotechnology, 2024 Feb 09;35(17).
PMID: 38262054 DOI: 10.1088/1361-6528/ad21a2

Heparins are a family of sulfated linear negatively charged polysaccharides that have been widely used for their anticoagulant, antithrombotic, antitumor, anti-inflammatory, and antiviral properties. Additionally, it has been used for acute cerebral infarction relief as well as other pharmacological actions. However, heparin's self-aggregated macrocomplex may reduce blood circulation time and induce life-threatening thrombocytopenia (HIT) complicating the use of heparins. Nonetheless, the conjugation of heparin to immuno-stealth biomolecules may overcome these obstacles. An immunostealth recombinant viral capsid protein (VP28) was expressed and conjugated with heparin to form a novel nanoparticle (VP28-heparin). VP28-heparin was characterized and tested to determine its immunogenicity, anticoagulation properties, effects on total platelet count, and risk of inducing HIT in animal models. The synthesized VP28-heparin trimeric nanoparticle was non-immunogenic, possessed an average hydrodynamic size (8.81 ± 0.58 nm) optimal for the evasion renal filtration and reticuloendothelial system uptake (hence prolonging circulating half-life). Additionally, VP28-heparin did not induce mouse death or reduce blood platelet count when administered at a high dosein vivo(hence reducing HIT risks). The VP28-heparin nanoparticle also exhibited superior anticoagulation properties (2.2× higher prothrombin time) and comparable activated partial thromboplastin time, but longer anticoagulation period when compared to unfractionated heparin. The anticoagulative effects of the VP28-heparin can also be reversed using protamine sulfate. Thus, VP28-heparin may be an effective and safe heparin derivative for therapeutic use.

Matched MeSH terms: Blood Coagulation
Coagulation studies in Asian women using injectable progestogen for contraception

Tsakok FH, Koh S, Yuen R, Ratnam SS

Int J Gynaecol Obstet, 1980 9 1;18(2):105-8.
PMID: 6108245 DOI: 10.1002/j.1879-3479.1980.tb00256.x

Coagulation, fibrinolytic activity and platelet function were studied in 104 Asian women volunteers who received 150 mg of depot medroxyprogesterone acetate intramuscularly every three months for two years or more. The results were compared with those in matched controls. There was a paucity of change in coagulation factors. The fibrinogen levels were increased and prothrombin time was shortened. The fibrinolytic activity, as shown by the euglobulin clot lysis time, was significantly increased. This latter change contrasts with the many reports concerning Caucasian women and may reflect an increase in fibrinolytic potential in Asian women.

Matched MeSH terms: Blood Coagulation/drug effects*; Blood Coagulation Factors/analysis; Blood Coagulation Tests
Fulltext Effects of plateletpheresis on blood coagulation parameters in healthy donors at National Blood Centre, Kuala Lumpur, Malaysia

Siti Nadiah AK, Nor Asiah M, Nur Syimah AT, Normi M, Anza E, Aini AN, et al.

Transfus Apher Sci, 2013 Dec;49(3):507-10.
PMID: 24055412 DOI: 10.1016/j.transci.2013.08.004

Plateletpheresis is a method used to remove platelet from the body either from random volunteer donors, patient's family members or HLA matched donors. A cross sectional study was carried out on 59 plateletpheresis donors aged between 18 and 55 years at National Blood Center (NBC), Kuala Lumpur. We compared the blood parameters before and after plateletpheresis and we found that the platelet count, FVIII, fibrinogen and thrombophilia markers anti-thrombin (AT), protein C and protein S were significantly reduced (p<0.05) with prolonged PT and APTT. There were significant changes in blood coagulation parameters but it is within acceptable range.

Matched MeSH terms: Blood Coagulation/physiology*
A comparative study on the biological properties of venoms from juvenile and adult common tiger snake (Notechis scutatus) venoms

Tan NH, Ponnudurai G, Mirtschin PJ

Comp. Biochem. Physiol., B, 1993 Nov;106(3):651-4.
PMID: 8281760

1. The biological properties of venoms from juvenile and adult common tiger snakes (Notechis scutatus) were compared. 2. The lethality, procoagulant activity and enzymatic activities of the juvenile venom were not substantially different from those of the adult venom. 3. Electrophoretic studies, however, indicated some minor differences in the protein composition of the juvenile and adult venoms.

Matched MeSH terms: Blood Coagulation Factors/analysis*

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