[Inventive activity of the Departments of Protein Structure and Function, and Molecular Immunology of the Palladin Institute of Biochemistry of NAS of Ukraine. Part II. National breakthrough in the study and diagnostics of human hemostasis system].

The scientists of Protein Structure and Function, and Molecular Immunology Departments of the Palladin Institute of Biochemistry (NAS of Ukraine) under the supervision of member of NASU and NAMSU, prof. S. V. Komisarenko and corresponding member of NASU prof. E. V. Lugovskoy have made the real breakthrough in the field of research of the mechanisms of fibrin polymerization and formation of fibrin framework of thrombi. The immunodiagnostic test-systems for the evaluation of the risk of thrombus formation were developed for the first time. Researches have obtained the monoclonal antibodies to fibrinogen, fibrin, D-dimer and their fragments. These monoclonal antibodies were used as molecular probes for the localization of newly detected fibrin polymerization sites. Obtained antibodies with high affinity interact with fibrinogen, D-dimer and soluble fibrin ­ main markers of the risk of thrombus formation. They were used for the development of the immunodiagnostic test-systems to quantify these markers in human blood plasma for the evaluation of the state of haemostasis system, detection of prethrombotic states, disseminated intravascular coagulation, detection of thrombosis and monitoring of antithrombotic and fibrinolytic therapy. The successful trial of developed test-systems was carried out in clinics of Ukraine, and the State registration was obtained for the implementation of them into the clinical practice. Presented works were awarded State prize of Ukraine in Science and technology.

[Inventive activity of the Departments of Chemistry and Biochemistry of Enzymes, and Protein Structure and Function of the Palladin Institute of Biochemistry of NAS of Ukraine. Part III. Diagnostic test-systems for analysis of fibrinolysis blood system and novel approaches to thrombosis treatment].

This article continues analysis of scientific achievements of the Institute of Biochemistry in the study of hemostasis system. Two previous articles were focused on the studies of blood coagulation proteins and development of the immune-enzyme test-systems for evaluation of the risk of thrombosis upon various pathologies. This article highlights the research on the blood fibrinolysis system and new approaches to thrombosis treatment, which were developed (and are under development) in the Palladin Institute of Biochemistry of the NAS of Ukraine, in particular, in the Department of Chemistry and Biochemistry of Enzymes headed previously by Dr.Sci.(Biol.) S. O. Kudinov and now by Dr.Sci.(Biol.) T .V. Grinenko, and also in the Department of Protein Structure and Function headed by Dr.Biol.Sci. E. M. Makogonenko. The fundamental knowledge of protein molecule functions and mechanisms of regulation of blood coagulation and fibrinolysis opens up new opportunities to diagnose hemostasis disorders and control the effectiveness of the cardiovascular disease treatment and also contributes to development of new techniques for isolation of new proteins ­ promising therapeutic agents.

The old and the new in prekallikrein deficiency: historical context and a family from Argentina with PK deficiency due to a new mutation (Arg541Gln) in exon 14 associated with a common polymorphysm (Asn124Ser) in exon 5.

Prekallikrein (PK) is one of the clotting factors involved in the contact phase of blood. PK has an important historical role as its deficiency state represents the second instance of a clotting defect without bleeding manifestations, the first one being factor XII deficiency. PK deficiency is a rare clotting disorder. Moreover, only 11 patients have been investigated so far by molecular biology techniques. In this article, we briefly review some of the history around PK and also present some recent data on a newly identified family from Argentina suffering from PK deficiency. Two patients are homozygous whereas other family members are heterozygous. PK activity and antigen are 1% of normal in the homozygotes and around 60 to 70% of normal in the heterozygotes. As expected, all patients are asymptomatic of bleeding or thrombosis presentations. However, the two homozygotes showed essential hypertension. The PK deficiency in this family is due to a new mutation (Arg541Gln) in exon 14. The defect segregates together with a known polymorphism, Asn124Ser, in exon 5. The significance of the presence of hypertension in the two homozygotes is discussed in view of the extra coagulation effects of PK on vasodilation, vessel permeability, and the control of blood pressure. Structure function analysis indicates that the substitution of Arg with Gln probably impedes the transmembrane diffusion of the molecule, which therefore cannot be secreted in the homozygotes. The presence of hypertension in patients with PK deficiency has been previously reported in some but not all patients. Future research activities will probably concentrate on the effect of PK and other contact phase factors on the vascular system.

Towards hemostatic resuscitation: the changing understanding of acute traumatic biology, massive bleeding, and damage-control resuscitation.

During the past decade there has been a profound change in the understanding of postinjury coagulation. Concurrently, new data suggest that a resuscitative strategy to minimize large volumes of crystalloid while recreating whole is associated with reduced morbidity and mortality. This article outlines the history of resuscitation and transfusion practices in trauma, the changing understanding of coagulation and inflammation, and clinical data driving changes in resuscitative conduct. Finally, the current state of the science suggests future basic science and clinical investigation that will drive changes in transfusion and resuscitation in severely injured military personnel and civilian patients.

The crucifixion of Jesus: review of hypothesized mechanisms of death and implications of shock and trauma-induced coagulopathy.

The crucifixion of Jesus is arguably the most well-known and controversial execution in history. Christian faithful, dating back to the time of Jesus, have believed that Jesus was executed by crucifixion and later returned physically to life again. Others have questioned whether Jesus actually died by crucifixion, at all. From review of medical literature, physicians have failed to agree on a specific mechanism of Jesus' death. A search of Medline/Pubmed was completed with respect to crucifixion, related topics, and proposed mechanisms of Jesus' death. Several hypotheses for the mechanism of Jesus' death have been presented in medical literature, including 1) Pulmonary embolism 2) Cardiac rupture 3) Suspension trauma 4) Asphyxiation 5) Fatal stab wound, and 6) Shock. Each proposed mechanism of Jesus' death will be reviewed. The events of Jesus' execution are described, as they are pertinent to development of shock. Traumatic shock complicated by trauma-induced coagulopathy is proposed as a contributing factor, and possibly the primary mechanism, of Jesus' death by crucifixion.

Hemostasis research in India: past, present, and future.

Hemostasis research in India has a long history considering the fact that it is one of the youngest specialities in the world. If we take creation of prothrombin time (PT) test as one of the beginning of modern hemostsis research, then the specialty is no older than 60 years. School of Tropical Medicine Kolkata, Banaras Hindu University, All India Institute of Medical Sciences at Delhi, Christian Medical College at Vellore, Post Graduate Institute of Medical Education and Research at Chandigarh, and KEM Hospital at Mumbai contributed substantially in defining various bleeding disorders in our country. Unfortunately, some of these institutes are no longer as active in the field as they used to be. Currently, the Institute of Immunohaematology at Mumbai, Chrstian medical College at Vellore, and All India Institute of Medical Sciences at Delhi are actively engaged in hemostsis research in India. Developing prenatal diagnostic technologies, mutation detection of various hemostatic disorders, developing low-cost management technologies for hemophilia, and other bleeding disorders are becoming important present day research activity in the area of hemostasis in addition to age old areas of prevalence and unusual case description studies. Entry of many new corporate hospitals, development of structured postgraduate training program in hematology, and easy availability of instruments and reagents are likely to foster further growth in this area of medical research in India in future.

Discovery of anticoagulant drugs: a historical perspective.

The history of the traditional anticoagulants is marked by both perseverance and serendipity. The anticoagulant effect of heparin was discovered by McLean in 1915, while he was searching for a procoagulant in dog liver. Link identified dicumarol from spoiled sweet clover hay in 1939 as the causal agent of the sweet clover disease, a hemorrhagic disorder in cattle. Hirudin extracts from the medicinal leech were first used for parenteral anticoagulation in the clinic in 1909, but their use was limited due to adverse effects and difficulties in achieving highly purified extracts. Heparins and coumarins (i.e.: warfarin, phenprocoumon, acenocoumarol) have been the mainstay of anticoagulant therapy for more than 60 years. Over the past decades, the drug discovery paradigm has shifted toward rational design following a target-based approach, in which specific proteins, or "targets", are chosen on current understandings of pathophysiology, small molecules that inhibit the target's activity may be identified by high-throughput screening and, in selected cases, these new molecules can be developed further as drugs. Despite the application of rational design, serendipity has still played a significant role in some of the new discoveries. This review will focus on the discovery of the main anticoagulant drugs in current clinical use, like unfractionated heparin, low-molecular-weight heparins, fondaparinux, coumarins (i.e.: warfarin, acenocoumarol, phenprocoumon), parenteral direct thrombin inhibitors (DTIs) (i.e.: argatroban, recombinant hirudins, bivalirudin), oral DTIs (i.e.: dabigatran) and oral direct factor Xa inhibitors (i.e.: rivaroxaban, apixaban).

Fifty-five years (1955-2010) of the Coagulation Section at Laboratory of Hematology, Sestre milosrdnice University Hospital, and its founder, hematologist Ljubomir Popovic.

The Coagulation Section at Laboratory of Hematology, Sestre milosrdnice University Hospital, Zagreb, was founded in 1955 by Ljubomir Popovic, hematologist and assistant at School of Medicine, University of Zagreb, in cooperation with hard-working laboratory technicians. Apart from papers on hematologic neoplasms, plasmacytoma and lymphoma, Ljubomir Popovic published a number of papers in the field of anticoagulant therapy with heparin and oral anticoagulants, some of which are also in use today. After Ljubomir Popovic left the Hospital in 1964, the Laboratory was run by Professor Nedjeljko Milic, head of the newly founded Division of Hematology. In 1968, the management of the Laboratory of Hematology was taken over by Biserka Raic, MS, medical biochemist, until her retirement in 2007. Great development in morphological and cytometric studies of blood and blood cells has been paralleled by continuous progress and almost dominating activities in the diagnosis of hemostasis disorders. In the 1970s, Marko Koprcina, hematologist, and Biserka Raic introduced the then latest tests in practice at all Hospital departments. In that golden age of the Coagulation Section, M. Koprcina, B. Raic and their associates transferred their knowledge to all colleagues in the Hospital. Through that collaboration, high standards in the diagnosis of hemostasis disorders were achieved, from which the currently high level of clinical knowledge about coagulation disorders and their treatment has derived, making Sestre milosrdnice University Hospital one of the leading hospitals in this field in the country. By describing development of the Coagulation Section and the life of its founder Ljubomir Popovic, the authors tried to provide an answer to the following question: can today's clinicians still have a deciding role in laboratory development, considering that assessments of different phenomena are always initiated by an interested clinician who is trying to interpret and understand the nature of the disorder? This means that the clinician's place may still be in the laboratory, or else, it will become clear that the laboratory, as well as knowledge in general, has undergone such an expansion that the clinician is no longer able to run it by himself. It is our belief that the answer will assert itself through the survey of the history of the Coagulation Section at Laboratory of Hematology, Division of Hematology, and the lives of its founders and beneficiaries.

Desmopressin (DDAVP) in bleeding disorders of childhood.

As in adults, desmopressin (DDAVP) can be used in children for prophylaxis of bleeding and to stop bleeding in many hereditary and acquired bleeding disorders. DDAVP is the treatment of choice in children with mild hemophilia and type 1 von Willebrand's disease (vWD). It is effective in some variants of vWD and in many patients with platelet function defects. It reduces the bleeding diathesis of children with uremia and drug-induced bleeding complications. In any case, a test dose of DDAVP has to be given to the patient to predict the hemostatic effect before relying on this drug for treatment. The response can be measured by shortening of the bleeding time (BT) and of partial thromboplastin time (PTT), indicating a rise of Factor (F) VIII or von Willebrand factor (vWF). Side effects such as facial flushing, transient headache, increased pulse rate, and drop in systolic blood pressure are mild and transient. They can be minimized when the dose is not exceeding 0.3 microg/kg body weight, and the infusion lasts at least 20 to 30 minutes. The strong antidiuretic action of DDAVP has some potential problems that are negligible in adults and older children when water intake is restricted. In infants and small children under the age of 18 months, however, DDAVP should be used with caution and with close surveillance in order to prevent water intoxication and electrolyte imbalance. The danger is increased when the patients receive parenteral fluid substitution. The advantages of DDAVP include the reduction in the use of plasma factor concentrates, thereby minimizing the danger of immunological or infectious complications, as well as the considerable reduction of costs realized by treatment with this form of medication. Fortunately, it can be applied successfully in the most frequent hereditary bleeding disorder, namely vWD type 1.

Blood clots: the nineteenth-century debate over the substance and means of transfusion in Britain.

Historians have devoted little attention to blood transfusion in the nineteenth century. In part, this neglect reflects the presentist assumption that, before Karl Landsteiner's discovery of blood types, this practice would have failed too often to gain currency. Yet, transfusion was in fact the subject of much debate, and was actively practised, primarily by obstetricians on haemorrhaging women. Examining this practice through the conceptual lens of 'blood clots', both as noun and as observation, I follow transfusors' assumptions about the nature of the blood and the problem of its coagulation. Tracing the medicalization of ideas about blood by the century's end, I map this shift onto changing notions about why transfusion was performed, what substance was best employed, and what instrument best fitted that substance's movement into the circulation. In this way, 'Blood Clots' reconstructs the discourse surrounding transfusion, extending that discourse to material culture in order to illuminate the rationale that guided transfusion's practice in nineteenth-century Britain.

Association of blood groups and disease: do blood group antigens and antibodies have a biological role?

Although there are probably over a thousand publications on the associations of blood groups and disease, many are based totally on statistical analyses. Most of the earlier studies have been controversial, because they were small studies and/or had inadequate controls and/or had been analyzed incorrectly. Nevertheless, it is difficult to argue with the general pattern that emerges from the large body of statistical data on malignancy, coagulation and infection. Recent findings in membrane chemistry, rumor immunology and infectious disease (especially relating to bacterial receptors), add a scientific rationale for some of these findings, and there is an increasing rationale for some of the earlier statistical findings. Some of the more recent findings on parasitic/bacterial/viral receptors, the hematological abnormalities seen when high frequency blood group antigens are missing, and the association with immunologically important proteins are most convincing and suggest that blood group antigens do sometimes play a biological role; this role may relate directly, or often be completely unrelated, to the red cell.