New diagnostic approaches for venous thromboembolism and thrombophilia

Background: Venous thromboembolism is a serious disease comprising both pulmonary embolism and deep vein thrombosis. Venous thromboembolism causes considerable mortality and morbidity, with residual symptoms such as pulmonary hypertension and post thrombotic syndrome. Current assays for venous thromboembolism and the predisposition to develop venous thromboembolisms (thrombophilia) can only reflect a fragment of the complicated hemostatic processes. This thesis evaluates the usefulness of current and emerging hemostatic assays that reflect several aspects of the hemostatic process. Increased diagnostic specificity would enable better diagnosis and risk stratification of patients with venous thromboembolism. Improved biomarkers for venous thromboembolism and thrombophilia have the potential to prevent major morbidity and mortality by guiding treatment and prophylaxis to the right patients at the right time. Aim: The aim of this project was to investigate assays that could improve the care for patients with suspected venous thromboembolism or thrombophilia. We evaluated the usefulness of current and emerging biomarkers for venous thromboembolism, explored the utility of emerging biomarkers for thrombophilia and functionally characterized a novel prothrombotic genetic variant. Methods: A prospective case-control study of 954 patients with clinically suspected acute deep venous thrombosis or pulmonary embolism were recruited from the emergency department and analyzed by four D-dimer assays, fibrin monomers, thrombin generation and fibrin aggregation assays. The discriminatory accuracy of all assays and of age-adjusted cutoffs for D-dimer was evaluated. From the special coagulation laboratory, we included 369 patients with clinical criteria for thrombophilia testing. Plasma and DNA samples were analyzed by the global hemostatic assays Overall Hemostatic Potential (OHP) and Endogenous Thrombin Potential (ETP) as well as genotyped for several prothrombotic variants. In a separate study, a novel genetic variant with possible prothrombotic effect was characterized by the same assays, prothrombin levels, mRNA expression, and scanning electron microscopy of fibrin clot structure. Conclusion: Use of age-adjusted cutoffs for D-dimer could lead to a >5% decrease in false positivity rate and in elderly patients this avoidance of unwarranted radiology could even affect as many as 20% of patients with suspected venous thromboembolism. The Overall Hemostatic Potential, Endogenous Thrombin Potential and fibrin monomer assay were not superior to D-dimer for diagnosis of deep venous thrombosis or pulmonary embolism. Global hemostatic assays and extended investigation of prothrombotic genetic variants discretely improved the predictive ability of the classical genetic thrombophilia markers and the proportion of patients with verifiable hypercoagulability. We could also suggest a connection between increased thrombotic risk and a recently discovered synonymous single nucleotide polymorphism

Markers of neutrophil activation and neutrophil extracellular traps in diagnosing patients with acute venous thromboembolism: A feasibility study based on two VTE cohorts

Background Venous thromboembolism (VTE) diagnosis would greatly benefit from the identification of novel biomarkers to complement D-dimer, a marker limited by low specificity. Neutrophil extracellular traps (NETs) have been shown to promote thrombosis and could hypothetically be used for diagnosis of acute VTE. Objectives To assess the levels of specific markers of neutrophil activation and NETs and compare their diagnostic accuracy to D-dimer. Methods We measured plasma levels of neutrophil activation marker neutrophil elastase (NE), the NET marker nucleosomal citrullinated histone H3 (H3Cit-DNA) and cell-free DNA in patients (n = 294) with suspected VTE (pulmonary embolism and deep vein thrombosis) as well as healthy controls (n = 30). A total of 112 VTE positive and 182 VTE negative patients from two prospective cohort studies were included. Results Higher levels of H3Cit-DNA and NE, but not cell-free DNA, were associated with VTE. Area under receiver operating curves (AUC) were 0.90 and 0.93 for D-dimer, 0.65 and 0.68 for NE and 0.60 and 0.67 for H3Cit-DNA in the respective cohorts. Adding NE and H3Cit-DNA to a D-dimer based risk model did not improve AUC. Conclusions Our study demonstrates the presence of neutrophil activation and NET formation in VTE using specific markers. However, the addition of NE or H3Cit-DNA to D-dimer did not improve the discrimination compared to D-dimer alone. This study provides information on the feasibility of using markers of NETs as diagnostic tools in acute VTE. Based on our findings, we believe the potential of these markers are limited in this setting

Federation of European Laboratory Animal Science Associations recommendations of best practices for the health management of ruminants and pigs used for scientific and educational purposes

Most ruminants and pigs used for scientific and educational aims are bred not for these purposes but in a farm environment. Given the wide range of diseases that these species might have, ensuring that the animals’ health status is appropriate can be complex and challenging. The Federation of European Laboratory Animal Science Associations has previously published recommendations for the health monitoring of experimental colonies of pigs (1998) and, respectively, calves, sheep and goats (2000). Unfortunately, the uptake of those recommendations was poor and insufficiently reported in scientific publications. These new recommendations for best practice focus on the main species of ruminants (cattle, sheep and goats) and pigs. They provide general and specific information helpful for designing a health management programme for the suppliers and for the user establishments, as well as guidance on animal procurement. Critical thinking based on the fields of use of the animals is promoted, aiming to help in taking informed decisions rather than establishing an exhaustive exclusion list for pathogens. Implementing the best health and welfare management practices should be done under the guidance of a competent attending veterinarian, with expertise and sufficient authority to take the appropriate action, doubled by excellent communication skills. It is strongly recommended that the user establishment’s veterinarian works in close collaboration with the supplier’s veterinarian. // Die meisten Wiederkäuer und Schweine, die zu wissenschaftlichen und Ausbildungszwecken dienen, werden nicht eigens dafür, sondern in einem landwirtschaftlichen Umfeld gezüchtet. Angesichts des breiten Spektrums potenzieller Krankheiten bei diesen Tierarten kann die Gewährleistung eines adäquaten Gesundheitszustands der Tiere durchaus komplex und schwierig sein. FELASA hat bereits früher Empfehlungen für die Gesundheitsüberwachung von Versuchskolonien von Schweinen (1998) bzw. Kälbern, Schafen und Ziegen (2000) veröffentlicht. Leider stießen diese Empfehlungen auf mangelndes Echo und wissenschaftliche Publikationen berichteten diesbezüglich nur unzureichend. Die vorliegenden neuen Empfehlungen für beste Praxis konzentrieren sich auf die wichtigsten Arten von Wiederkäuern (Rinder, Schafe und Ziegen) sowie auf Schweine. Sie enthalten allgemeine und spezifische Informationen, die für die Gestaltung eines Gesundheitsmanagementprogramms für die Lieferanten und für die Verwendereinrichtungen nützlich sind, ebenso wie Hinweise zur Tierbeschaffung. Kritisches Denken auf der Grundlage der Anwendungsbereiche der Tiere, das darauf abzielt, fundierte Entscheidungen zu treffen, anstatt eine erschöpfende Ausschlussliste für Krankheitserreger zu erstellen, wird unterstützt. Die Umsetzung der besten Gesundheits- und Tierschutzmanagementpraktiken sollte unter der Anleitung eines kompetenten behandelnden Tierarztes erfolgen, der über Fachwissen und genügend Autorität sowie über ausgezeichnete Kommunikationsfähigkeiten verfügen, um die entsprechenden Maßnahmen zu ergreifen und zu vermitteln. Eine enge Zusammenarbeit zwischen dem Tierarzt der Verwendereinrichtung und dem Tierarzt des Lieferanten wird dringend empfohlen

Presence of a Gender Perspective in Psychotherapeutic Assesments : A Qualitative Study

 Föreliggande studie avsåg att undersöka förekomst av genusperspektiv i klientarbete hos yrkesverksamma terapeuter. Sex stycken semi-strukturerade intervjuer om ett fiktivt kvinnligt klientcase genomfördes med väl etablerade terapeuter med minst steg1- utbildning i KBT. På grund av risken för socialt önskvärda svar blindades terapeuterna initialt för den del av syftet som rörde genus. Studien hade en induktiv ansats och materialet analyserades med tematisk analys. Analysen ledde till fyra huvudteman: Att vara kvinna och mamma är att vara stressad, Gilla läget, Jämställdhet som valfri livsstil – upp till varje person  samt Feministisk medvetenhet.  Resultaten visar att strukturella problem ofta hanteras med individualterapeutiska åtgärder. Jämställdhet mellan könen konceptualiseras som ett personligt val och den feministiska medvetenheten hos terapeuterna varierar. Ett genomfört maktkritiskt synsätt i både bedömning och behandlingsförslag finns inte. Terapeuterna uttrycker kunskap om och intresse för genus som något vid sidan av det psykoterapeutiska arbetet. Studiens resultat talar för behov av mer utbildning om genus på psykoterapeututbildningar

Causes of death after first time venous thromboembolism

Abstract Background Causes of death after first time community-acquired venous thromboembolism (VTE) diagnosed in unselected patients at the emergency department (ED) was investigated. Materials and methods The study consists of all patients > 18 years of age who had a visit for any medical reason to any of 5 different ED in Stockholm County, Sweden from 1st January 2016 to 31st December 2017. We have identified all patients with a first registered incident VTE; deep vein thrombosis (DVT) and/or pulmonary embolism (PE) during the study period. Cox regression models were used to estimate hazards ratios (HR) with 95% confidence intervals (CIs) for all-cause mortality and cause-specific death in patients with DVT or PE using all other patients as the reference group. Results In total, 359,884 patients had an ED visit during the study period of whom about 2.1% were diagnosed with VTE (DVT = 4,384, PE = 3,212). The patients with VTE were older compared to the control group. During a mean follow up of 2.1 years, 1567 (21%) and 23,741(6.7%) patients died within the VTE and reference group, respectively. The adjusted risk of all-cause mortality was nearly double in patients with DVT (HR 1.7; 95% CI, 1.5–1.8) and more than 3-fold in patients with PE (HR 3.4; 95% CI, 3.1–3.6). While the risk of cancer related death was nearly 3-fold in patient with DVT (HR 2.7; 95% CI, 2.4–3.1), and 5-fold in PE (HR 5.4; 95% CI, 4.9-6.0 respectively). The diagnosis of PE during the ED visit was associated with a significantly higher risk of cardiovascular death (HR 2.2; 95% CI, 1.9–2.6). Conclusion Patients with VTE have an elevated risk of all-cause mortality, including cardiovascular death

Diagnostic Accuracy in Acute Venous Thromboembolism: Comparing D-Dimer, Thrombin Generation, Overall Hemostatic Potential, and Fibrin Monomers

Introduction For acute venous thromboembolism (VTE), a biomarker with higher specificity than D-dimer would be of great clinical use. Thrombin generation and overall hemostatic potential (OHP) reflect the hemostatic balance by globally assessing multiple coagulation factors and inhibitors. These tests discriminate between healthy controls and patients with a prothrombotic tendency but have yet to be established as clinical biomarkers of VTE. Objective This study compares endogenous thrombin potential (ETP) and OHP to D-dimer and fibrin monomers (FM) in outpatients with suspected VTE. Methods A cross-sectional diagnostic study where 954 patients with suspected pulmonary embolism or deep venous thrombosis were recruited consecutively from the medical emergency department at Karolinska University Hospital. D-dimer, FM, OHP, and ETP were analyzed in a subpopulation of 60 patients with VTE and 98 matched controls without VTE. VTE was verified either by ultrasonography or computed tomography and clinical data were collected from medical records. Results Compared with healthy controls, both VTE and non-VTE patients displayed prothrombotic profiles in OHP and ETP. D-dimer, FM, ETP area under the curve (AUC), and ETP T lag were significantly different between patients with VTE and non-VTE. The largest receiver-operating characteristic AUCs for discrimination between VTE and non-VTE, were found in D-dimer with 0.94, FM 0.77, and ETP AUC 0.65. No useful cutoff could be identified for the ETP or the OHP assay. Conclusion Compared with D-dimer, neither ETP nor OHP were clinically viable biomarkers of acute venous thrombosis. The data indicated that a large portion of the emergency patients with suspected VTE were in a prothrombotic state.Funding agencies: Foundation for Coagulation Research at Karolinska Institutet, the ScandinavianResearch Foundation for Varicose Veins and other Venous Diseases, FoU Region Stockholm, the Swedish Society onThrombosis and Haemostasis with Leo Pharma.</p

The Silence Speaks, but We Do Not Listen: Synonymous c.1824C gt T Gene Variant in the Last Exon of the Prothrombin Gene as a New Prothrombotic Risk Factor

BACKGROUND: Thrombosis is a major global disease burden with almost 60% of cases related to underlying heredity and most cases still idiopathic. Synonymous single nucleotide polymorphisms (sSNPs) are considered silent and phenotypically neutral. Our previous study revealed a novel synonymous FII c.1824C gt T variant as a potential risk factor for pregnancy loss, but it has not yet been associated with thrombotic diseases. METHODS: To determine the frequency of the FII c.1824C gt T variant we have sequenced patients' DNA. Prothrombin RNA expression was measured by quantitative PCR. Functional analyses included routine hemostasis tests, western blotting and ELISA to determine prothrombin levels in plasma, and global hemostasis assays for thrombin and fibrin generation in carriers of the FII c.1824C gt T variant. Scanning electron mi- croscopy was used to examine the structure of fibrin clots. RESULTS: Frequency of the FII c.1824C gt T variant was significantly increased in patients with venous thromboembolism and cerebrovascular insult. Examination in vitro demonstrated increased expression of prothrombin mRNA in FII c.1824C gt T transfected cells. Our ex vivo study of FII c.1824C gt T carriers showed that the presence of this variant was associated with hyperprothrom-binemia, hypofibrinolysis, and formation of densely packed fibrin clots resistant to fibrinolysis. CONCLUSION: Our data indicate that FII c.1824C gt T, although a synonymous variant, leads to the development of a prothrombotic phenotype and could represent a new prothrombotic risk factor. As a silent variant, FII c.1824C gt T would probably be overlooked during genetic screening, and our results show that it could not be detected in routine laboratory tests

POLYPHENO srl - Spin Off Accademica Accreditata del''Università degli Studi di Bari "Aldo Moro"

La società ha per oggetto le seguenti attività: 1. Consulenza per finalità di industrializzazione di materiali e prodotti innovativi in campo sanitario attraverso lo sviluppo di attività per l’ottenimento di materie prime in ambito nutraceutico; 2. Consulenza per lo sviluppo di studi di farmacoeconomia; 3. Consulenza per sviluppo di studi sui rapporti tra ambiente, nutrizione e salute; 4. Fund raising ad impatto sociale