Efficiency of Neat and Quaternized-Cellulose Nanofibril Fillers in Chitosan Membranes for Direct Ethanol Fuel Cells.

In this work, fully polysaccharide based membranes were presented as self-standing, solid polyelectrolytes for application in anion exchange membrane fuel cells (AEMFCs). For this purpose, cellulose nanofibrils (CNFs) were modified successfully with an organosilane reagent, resulting in quaternized CNFs (CNF (D)), as shown by Fourier Transform Infrared Spectroscopy (FTIR), Carbon-13 (C13) nuclear magnetic resonance (13C NMR), Thermogravimetric Analysis (TGA)/Differential Scanning Calorimetry (DSC), and ζ-potential measurements. Both the neat (CNF) and CNF(D) particles were incorporated in situ into the chitosan (CS) membrane during the solvent casting process, resulting in composite membranes that were studied extensively for morphology, potassium hydroxide (KOH) uptake and swelling ratio, ethanol (EtOH) permeability, mechanical properties, ionic conductivity, and cell performance. The results showed higher Young's modulus (119%), tensile strength (91%), ion exchange capacity (177%), and ionic conductivity (33%) of the CS-based membranes compared to the commercial Fumatech membrane. The addition of CNF filler improved the thermal stability of the CS membranes and reduced the overall mass loss. The CNF (D) filler provided the lowest (4.23 × 10-5 cm2 s-1) EtOH permeability of the respective membrane, which is in the same range as that of the commercial membrane (3.47 × 10-5 cm2s-1). The most significant improvement (~78%) in power density at 80 °C was observed for the CS membrane with neat CNF compared to the commercial Fumatech membrane (62.4 mW cm-2 vs. 35.1 mW cm-2). Fuel cell tests showed that all CS-based anion exchange membranes (AEMs) exhibited higher maximum power densities than the commercial AEMs at 25 °C and 60 °C with humidified or non-humidified oxygen, demonstrating their potential for low-temperature direct ethanol fuel cell (DEFC) applications.

Evaluation of Sediments from the River Drava and Their Potential for Further Use in the Building Sector.

Sedimentation is a naturally occurring process of allowing particles in water bodies to settle out of the suspension under a gravity effect. In this study, the sediments of the Drava River were fully investigated to determine the heavy metal concentrations along the river and their potential reuse in the construction sector. Naturally dehydrated sediments from the Drava River were tested as an additive for the production of fired bricks. The dredged sediments were used as a substitute for natural brick clay in amounts up to 50% by weight, and it was confirmed that up to 20% by weight of the added sediment could be used directly in the process without critically affecting performance. Finally, the naturally dehydrated sediments were also evaluated for their use as a filling material in the construction of levees. The natural moisture content of the dehydrated sediment was too high for it to be used without additives, so quicklime was added as an inorganic binder. The test results showed an improvement in the geotechnical properties of the material to such an extent that it is suitable as a filling material for levees.

Efficient Chitosan/Nitrogen-Doped Reduced Graphene Oxide Composite Membranes for Direct Alkaline Ethanol Fuel Cells.

Herein, we prepared a series of nanocomposite membranes based on chitosan (CS) and three compositionally and structurally different N-doped graphene derivatives. Two-dimensional (2D) and quasi 1D N-doped reduced graphene oxides (N-rGO) and nanoribbons (N-rGONRs), as well as 3D porous N-doped graphitic polyenaminone particles (N-pEAO), were synthesized and characterized fully to confirm their graphitic structure, morphology, and nitrogen (pyridinic, pyrrolic, and quaternary or graphitic) group contents. The largest (0.07%) loading of N-doped graphene derivatives impacted the morphology of the CS membrane significantly, reducing the crystallinity, tensile properties, and the KOH uptake, and increasing (by almost 10-fold) the ethanol permeability. Within direct alkaline ethanol test cells, it was found that CS/N rGONRs (0.07 %) membrane (Pmax. = 3.7 mWcm-2) outperformed the pristine CS membrane significantly (Pmax. = 2.2 mWcm-2), suggesting the potential of the newly proposed membranes for application in direct ethanol fuel cells.

Consolidation of cellulose nanofibrils with lignosulphonate bio-waste into excellent flame retardant and UV blocking membranes.

The use of biomass to produce value-adding materials is a core objective of the circular economy, which has attracted great research interest in recent decades. In this context, we present here a simple dispersion-casting process for consolidation of cellulose nanofibrils (CNF), lignosulphonate (LS)-rich bio-waste and CaCl2 in composite membranes. The addition of CaCl2 to CNF and LS dispersions reduces the ζ potential, due to an electrostatic screening, which promotes the aggregation of CNF, increases its moisture content and promotes LS deposition on CNFs already in the dispersion phase. Addition of both the LS and CaCl2 to CNF dispersion has an adverse effect on the mechanical properties of the final membranes. The effectiveness of the new composite membranes has been described in terms of their passive (charring) flame retardancy and 100 % UVA/UVB shielding capacity, both identified for membranes with the highest LS content, as well as high electronic resistance.

Functional dextran amino acid ester particles derived from N-protected S-trityl-L-cysteine.

This work describes the derivatization of dextran using N-(tert-butyloxycarbonyl)-S-(trityl)-L-cysteine in the presence of N,N'-carbonyldiimidazole (CDI) as a coupling agent. Homogeneous reactions in dimethyl sulfoxide allowed for an efficient coupling of the amino acid derivative to the polymer backbone. Derivatization was confirmed by infrared and 13C NMR spectroscopy, size exclusion chromatography and elemental analysis. The presence of hydrophobic protecting groups resulted in a product that can be shaped into water-insoluble particles stable in an aqueous environment and non-toxic for lung epithelial cells. It is suggested that materials composed of ester bonds between amino acids and polysaccharides are useful for targeted drug delivery, bio-imaging or surface functionalization.

Enrichment of enzymatically mineralized gellan gum hydrogels with phlorotannin-rich Ecklonia cava extract Seanol(®) to endow antibacterial properties and promote mineralization.

Hydrogels offer several advantages as biomaterials for bone regeneration, including ease of incorporation of soluble substances such as mineralization-promoting enzymes and antibacterial agents. Mineralization with calcium phosphate (CaP) increases bioactivity, while antibacterial activity reduces the risk of infection. Here, gellan gum (GG) hydrogels were enriched with alkaline phosphatase (ALP) and/or Seanol(®), a seaweed extract rich in phlorotannins (brown algae-derived polyphenols), to induce mineralization with CaP and increase antibacterial activity, respectively. The sample groups were unmineralized hydrogels, denoted as GG, GG/ALP, GG/Seanol and GG/Seanol/ALP, and hydrogels incubated in mineralization medium (0.1 M calcium glycerophosphate), denoted as GG/ALP_min, GG/Seanol_min and GG/Seanol/ALP_min. Seanol(®) enhanced mineralization with CaP and also increased compressive modulus. Seanol(®) and ALP interacted in a non-covalent manner. Release of Seanol(®) occurred in a burst phase and was impeded by ALP-mediated mineralization. Groups GG/Seanol and GG/ALP/Seanol exhibited antibacterial activity against methicillin-resistant Staphylococcus aureus. GG/Seanol/ALP_min, but not GG/Seanol_min, retained some antibacterial activity. Eluates taken from groups GG/ALP_min, GG/Seanol_min and GG/ALP/Seanol_min displayed comparable cytotoxicity towards MG-63 osteoblast-like cells. These results suggest that enrichment of hydrogel biomaterials with phlorotannin-rich extracts is a promising strategy to increase mineralizability and antibacterial activity.

Enhanced catalytic activity of the surface modified TiO2-MWCNT nanocomposites under visible light.

Fusing multiwall carbon nanotubes (MWCNTs) with TiO2 at the nano-scale level promotes the separation of those electron-hole charges generated upon UV and daylight irradiation. In this study, we investigated facile sonochemical synthesis, combined with the calcination process for the preparations of TiO2-MWCNT composites with different mole ratios of titanium and carbon. In order to produce stable nano dispersions we exploited an innovative biotechnology-based approach for the covalent functionalizations of TiO2-MWCNTs with in-situ synthesized soluble phenoxazine dye molecules. The none and functionalized TiO2-MWCNTs composites were analyzed by a range of analytical techniques including XRD, Raman, XPS, SEM and UV-vis diffuse reflectance spectroscopy (DRS), and dynamic light scattering (DLS). The photocatalytic activity was evaluated toward the liquid-phase degradation of MB in aqueous solution under both UV and visible light irradiation. TiO2-MWCNTs with optimized mole ratio exhibit much higher photocatalytic activity and stability than bare TiO2. The as-prepared TiO2-MWCNTs photocatalyst possessed good adsorptivity of dyes, extended light absorption range and efficient charge separation properties simultaneously. The results indicated that the soluble phenoxazine dyes and amino-benzenesulfonic acid monomers were covalently grafted on to the surfaces of TiO2-MCNTs, which promoted good aquatic dispersibility and extended light absorption, resulting in increased photocatalytic efficiency.

Chitosan hydrogels enriched with polyphenols: Antibacterial activity, cell adhesion and growth and mineralization.

Injectable hydrogels for bone regeneration consisting of chitosan, sodium beta-glycerophosphate (Na-ß-GP) and alkaline phosphatase (ALP) were enriched with the polyphenols phloroglucinol (PG) and gallic acid (GA) and characterized physicochemically and biologically with respect to properties relevant for applications in bone regeneration, namely gelation kinetics, mineralizability, antioxidant properties, antibacterial activity, cytocompatibility and ability to support adhesion and growth of human osteoblast-like MG63 cells. Enrichment with PG and GA had no negative effect on gelation kinetics and mineralizability. PG and GA both enhanced antioxidant activity of unmineralized hydrogels. Mineralization reduced antioxidant activity of hydrogels containing GA. Hydrogels containing GA, PG and without polyphenols reduced colony forming ability of Escherichia coli after 1h, 3h and 6h incubation and slowed E. coli growth in liquid culture for 150min. Hydrogels containing GA were cytotoxic and supported cell growth more poorly than polyphenol-free hydrogels. PG had no negative effect on cell adhesion and growth.

Characterisation and properties of homo- and heterogenously phosphorylated nanocellulose.

Nano-sized cellulose ester derivatives having phosphoryl side groups were synthesised by phosphorylation of nanofibrilated cellulose (NFC) and nanocrystaline cellulose (NCC), using different heterogeneous (in water) and homogeneous (in molten urea) processes with phosphoric acid as phosphoryl donor. The phosphorylation mechanism, efficacy, stability, as well as its influence on the NC crystallinity and thermal properties, were evaluated using ATR-FTIR and (13)C NMR spectroscopies, potentiometric titration, capillary electrophoresis, X-ray diffraction, colorimetry, thermogravimmetry and SEM. Phosphorylation under both processes created dibasic phosphate and monobasic tautomeric phosphite groups at C6 and C3 positioned hydroxyls of cellulose, yielded 60-fold (∼1,173 mmol/kg) and 2-fold (∼1.038 mmol/kg) higher surface charge density for p-NFC and p-NCC, respectively, under homogenous conditions. None of the phosphorylations affected neither the NC crystallinity degree nor the structure, and noticeably preventing the derivatives from weight loss during the pyrolysis process. The p-NC showed high hydrolytic stability to water at all pH mediums. Reusing of the treatment bath was examined after the heterogeneous process.

New findings about the lipase acetylation of nanofibrillated cellulose using acetic anhydride as acyl donor.

The acetylation efficiency of nanofibrillated cellulose (NFC) with acetic anhydride as acetyl donor was studied using lipase from Aspergillus niger in a mixture of dimethyl sulphoxide (DMSO) and phosphate buffer solution at ambient conditions and in supercritical carbon dioxide (scCO2). The chemical acetylation of NFC with comparable ester content was carried out for comparison. The ATR-FTIR, solid-state CP/MAS (13)C NMR and DSC analyses revealed that, besides the enzyme-catalysed acetylation, predominantly appearing at the C-6 position of cellulose hydroxyls, a strong and stable acyl-enzyme intermediate attachment also occurred on the NFC via Maillard reaction. Enzymatic acetylation via attached acyl-enzyme complex on NFC yielded high hydophobicity (contact angle of 84±9°), whereas the chemical acetylation with comparable ester content resulted in a much lower hydrophobic surface with a contact angle of 33±3°. Finally, the adsorption capacity profiles of lysozyme and BSA proteins on native, chemically and enzymatically acetylated NFC as a function of the pH medium were determined.

Nanocelluloses and their phosphorylated derivatives for selective adsorption of Ag(+), Cu(2+) and Fe(3+) from industrial effluents.

The potential of nanoscaled cellulose and enzymatically phosphorylated derivatives as bio-adsorbents to remove metal ions (Ag(+), Cu(2+) and Fe(3+)) from model water and industrial effluents is demonstrated. Introduction of phosphate groups onto nanocelluloses significantly improved the metal sorption velocity and sorption capacity. The removal efficiency was considered to be driven by the high surface area of these nanomaterials as well as the nature and density of functional groups on the nanocellulose surface. Generally, in the solutions containing only single types of metal ions, the metal ion selectivity was in the order Ag(+)>Cu(2+)>Fe(3+), while in the case of mixtures of ions, the order changed to Ag(+)>Fe(3+)>Cu(2+), irrespective of the surface functionality of the nanocellulose. In the case of industrial effluent from the mirror making industry, 99% removal of Cu(2+) and Fe(3+) by phosphorylated nanocellulose was observed. The study showed that phosphorylated nanocelluloses are highly efficient biomaterials for scavenging multiple metal ions, simultaneously, from industrial effluents.

Homogeneous and heterogeneous methods for laccase-mediated functionalization of chitosan by tannic acid and quercetin.

Homogeneous and heterogeneous methods for functionalization of chitosan with quercetin or tannic acid using laccase from Trametes versicolor is presented, yielding a bio-based product with synergistic antioxidant and antimicrobial properties. HPLC-SEC analysis and cyclic voltammetry kinetic studies showed that laccase catalyzes the oxidation of quercetin to electrophilic o-quinones, which undergo to an oligomer/polymer-forming structures. On the other hand, tannic acid was converted into gallic acid, its dimers, partially gallic acid esterified glucose and glucose, when exposed to laccases. ATR-FTIR spectroscopy provided evidence that quercetin o-quinones undergo coupling reactions with amino groups of chitosan via Schiff-base and Michael addition mechanisms under heterogeneous method, while oxidized tannic acid cross-linked with chitosan by hydrogen and electrostatic interactions under both methods. All polyphenols functionalized chitosans showed greatly improved ABTS(+) cation radicals scavenging capacity, compared with the untreated chitosan, while antimicrobial activities significantly depended on the mode of functionalization and type of microorganism.

Enzymatic reduction of complex redox dyes using NADH-dependent reductase from Bacillus subtilis coupled with cofactor regeneration.

Conventional vat dyeing involves chemical reduction of dyes into their water-soluble leuco form generating considerable amounts of toxic chemicals in effluents. In the present study, a new beta-nicotinamide adenine dinucleotide disodium salt (NADH)-dependent reductase isolated from Bacillus subtilis was used to reduce the redox dyes CI Acid Blue 74, CI Natural Orange 6, and CI Vat Blue 1 into their water-soluble leuco form. Enzymatic reduction was optimized in relation to pH and temperature conditions. The reductase was able to reduce Acid Blue 74 and Natural Orange 6 in the presence of the stoichiometrically consumed cofactor NADH; meanwhile, Vat Blue 1 required the presence of mediator 1,8-dihydroxyanthraquinone. Oxygen from air was used to reoxidize the dyes into their initial forms. The enzymatic reduction of the dyes was studied and the kinetic constants determined, and these were compared to the chemically-reduced leuco form. The enzyme responsible for the reduction showed homology to a NADH-dependent reductase from B. subtilis based on results from the MS/MS peptide mass mapping of the tryptically digested protein. Additionally, the reduction of Acid Blue 74 to its leuco form by reductase from B. subtilis was confirmed using NADH regenerated by the oxidation of formic acid with formate dehydrogenase from Candida boidinii in the same solution.

Effect of replacement fluids saline, gelofusine, and blood on biochemical and hematological parameters in rats subjected to repeated blood sampling.

BACKGROUND: Multiple blood sampling in rat pharmacokinetic (PK) and toxicokinetic studies can result in a pathophysiological response and misleading data interpretation. This study investigated whether these drawbacks can be minimized by replacing withdrawn blood with saline, gelofusine, or donor blood. MATERIAL/METHODS: Rats with cannulated jugular veins were subjected to 12 withdrawals of 250 and 500 microl of blood followed by replacement with the same amount of saline, gelofusine, or donor blood. Red blood cell, hematocrit, hemoglobin, total protein, albumin, glucose, Na, and Cl levels were determined in the drawn blood. RESULTS: Multiple blood sampling caused a time-dependent decrease in red blood cell counts and hematocrit, hemoglobin, total protein, and albumin levels in all the treatment groups, but these effects were significantly alleviated in the blood-treated group. Withdrawal of 500 microl of blood caused more pronounced changes than 250 microl blood withdrawal in the saline- and gelofusine-treatment groups, but not in the blood-treatment group. Plasma glucose and Na remained unchanged in all the treatment groups, while slight changes in Cl levels were observed in the saline- and gelofusine-treatment groups. CONCLUSIONS: Blood sample volume had a crucial impact on the measured parameters, which were least affected in the donor blood-treated group. Regardless of the treatment, up to five 250 microl blood samples can be drawn without causing significant changes in a rat's normal condition. However, for a proper determination of PK profile, more than five blood samples are required and therefore drawn blood should be replaced, preferably with donor blood.

Major and potential prothrombotic genotypes in patients with venous thrombosis and in healthy subjects from Slovenia.

The objective of our study was to investigate the prevalence of the polymorphisms factor V Leiden (FVL), prothrombin G20210A (PT G20210A), methylenetetrahydrofolate reductase C677T (MTHFR C677T), plasminogen activator inhibitor type 1 -675 4G/5G (PAI-1 4G/5G) and factor XII -4 C/T (FXII -4 C/T) in 295 Slovenian patients with venous thrombosis (VT) and 223 healthy controls in order to establish their contribution to the risk for VT. The major genetic risk factor was FVL, while PT G20210A, MTHFR 677 C/T, PAI-1 4G/5G and FXII -4 C/T polymorphisms were not. However, PT G20210A increased the risk of recurrent VT, MTHFR C677T increased the risk in older patients, while the FXII -4 T allele suggested a possible protective effect in younger patients. The risk of VT increased with increasing number of genetic defects.

Hemostasis activation in thrombophilic subjects with or without a history of venous thrombosis.

Thrombophilia is considered to increase the risk of venous thrombosis (VT) due to hemostasis activation. To determine the level of hemostasis activation in thrombophilic subjects with or without a history of VT, hemostasis activation markers prothrombin fragment 1 and 2 (F1+2), thrombin-antithrombin complex (TAT), and cross-linked fibrin degradation products (D-dimer) were measured in 94 subjects with (patients) and 101 subjects without a history of VT (controls). A total of 34.8% of patients and 14.8% of controls (P= .002) had at least 1 thrombophilic defect (protein C deficiency, activated protein C [APC] resistance, presence of lupus anticoagulants, or prothrombin G20210A polymorphism). The subjects were divided into 4 subgroups: patients with (TF(+) patients) and without (TF(-) patients) thrombophilia, and controls with (TF(+) controls) and without (TF(-) controls) thrombophilia. Hemostasis activation was comparable between all patients and controls (TAT: 2.1 vs 2.6 microg/L; F1+2: 1.0 vs 0.9 nmol/L; D-dimer: 36 vs 37 microg/L, respectively) and between TF(+) and TF(- ) patients. However, TF(+) controls had a significantly higher prevalence of increased hemostasis activation markers compared with TF(-) controls (TAT>4.4 microg/L, 38.4 vs 7.3%; F1+2>1.1 nmol/L, 53.8 vs 22.0%; D-dimer >78 microg/L, 30.7 vs 8.8% of subjects, respectively; all P< .05). After stratification for thrombophilic defects, hemostasis activation was associated with APC resistance in controls and with protein C deficiency in patients. To conclude, thrombophilia was associated with hemostasis activation in controls. We assumed that, in patients, the differences in hemostasis activation between subjects with or without thrombophilia were blurred due to undetermined and unidentified thrombophilic defects.

Utility of PFA-100 closure time vs. optical aggregometry in assessing the efficacy of platelet membrane glycoprotein IIb/IIIa antagonists in vitro.

BACKGROUND: Closure time measured by a platelet function analyser (PFA-100) was examined for its usefulness in assessing the efficacy of platelet membrane glycoprotein IIb/IIIa antagonists in vitro, and was compared to optical platelet aggregometry. METHODS: Three known glycoprotein IIb/IIIa antagonists [H-Arg-Gly-Asp-Ser-OH (RGDS), tirofiban and eptifibatide] and six new peptidomimetic glycoprotein IIb/IIIa antagonists (DKT-59, DPS-172, SMA-101, SMA-104, SMA-179 and SKN-191) were assessed. The concentration of antagonist which doubled closure time in collagen/ADP and collagen/epinephrine cartridges (IC200) or decreased ADP- or collagen-induced platelet aggregation by 50% (IC50) was used to assess the efficacy of the glycoprotein IIb/IIIa antagonist in inhibiting platelet function. RESULTS: IC200 for collagen/ADP and collagen/epinephrine closure times and IC50 for ADP- and collagen-induced platelet aggregation were highly associated (correlation coefficients 0.97-1.00, all p<0.001). Therefore, according to both methods, the most efficient glycoprotein IIb/IIIa antagonist was tirofiban (IC200=0.030-0.034 micromol/L, IC50=0.005-0.027 micromol/L) and the least efficient was RGDS (IC200=875-1100 micromol/L, IC50=124-377 micromol/L; all data are means), while the new peptidomimetic glycoprotein IIb/IIIa antagonists exhibited intermediate efficacies. CONCLUSIONS: Closure time represents a fast, simple and sensitive method of assessing glycoprotein IIb/IIIa antagonism in vitro, is comparable to optical aggregometry, and suitable for testing larger numbers of glycoprotein IIb/IIIa antagonists.

Evaluation of pre-analytical, demographic, behavioural and metabolic variables on fibrinolysis and haemostasis activation markers utilised to assess hypercoagulability.

BACKGROUND: Measurement of some haemostatic factors and products formed during activation of haemostasis seems to be promising in the determination of hypercoagulability. METHODS: The fibrinolytic variables euglobulin clot lysis time, tissue-type plasminogen activator, plasminogen activator inhibitor-1 and the haemostasis activation markers prothrombin fragment 1+2, thrombin-antithrombin complex and D-dimer were determined in 101 apparently healthy men and women aged 20-92 years (58+/-18 years, mean+/-SD) to establish variability due to several demographic, behavioural and metabolic factors. RESULTS: None of the fibrinolytic variables were affected by smoking, while tissue-type plasminogen activator antigen was significantly lower in women compared to men. Multiple regression analysis revealed several independent associations between tissue-type plasminogen activator, plasminogen activator inhibitor, body mass index and lipid levels, describing up to 40% of the variance in fibrinolytic variables. For haemostasis activation markers, no gender difference or effect of smoking was observed. Only D-dimer was independently associated with age. The haemostasis activation markers determined proved to be extremely sensitive to blood sampling procedure and were significantly higher in samples obtained by an untrained nurse compared to a trained nurse. CONCLUSIONS: Fibrinolytic variables are predominantly modulated by age, body mass index and blood lipids, while haemostasis activation markers are mainly un-influenced by these factors.

Evidence of hypercoagulability and inflammation in young patients long after acute cerebral ischaemia.

INTRODUCTION: Young subjects with acute cerebral ischaemia - stroke or transient ischaemic attack - form an etiologically heterogeneous and often not clearly explained group of patients. The aim was to investigate possible disturbances in haemostasis and inflammation long after an acute cerebral ischaemic event. MATERIALS AND METHODS: Forty-four consecutive patients referred after having suffered from acute cerebral ischaemia before the age of 45 participated 1 to 9 years (median value 5 years) after the event. At the time of blood sampling 33 (75%) patients were receiving antithrombotic treatment. Forty-six apparently healthy subjects of the same age group served as controls. In all subjects global haemostasis parameters (overall haemostasis, coagulation and fibrinolytic potential), thrombophilia, several markers of haemostasis activation and inflammation were determined. RESULTS: Patients did not differ from controls in most of the conventional risk factors and the presence of most forms of thrombophilia, although in seven (17.5%) patients the weak presence of lupus anticoagulants was observed. Patients had significantly increased overall haemostasis and coagulation potential, increased soluble P-selectin and D-dimer, decreased overall fibrinolysis potential and increased fibrinogen and C-reactive protein compared to controls. The subgroups of patients receiving antiplatelet treatment, with thrombophilia and recurrent acute cerebral ischaemia, did not differ significantly from the other patients. CONCLUSIONS: In young patients long after acute cerebral ischaemia an imbalance in the haemostatic system and a minor, but significant degree of inflammation was detected. The mechanisms behind haemostatic imbalance seem to be enhanced thrombin generation, platelet activation and depressed fibrinolysis.