Differences in morphology of fibrin clots induced with thrombin and ferric ions and its pathophysiological consequences.

The activation of blood coagulation leads to the formation of thrombin that, in turn, converts soluble plasma fibrinogen into insoluble fibrin clot. In healthy individuals, fibrin is effectively degraded; however, in prothrombotic states, proteolysis of fibrin clots are often delayed or even inhibited, and is associated with altered fibrin structure. We have previously shown that in inflammatory conditions like stroke and diabetes, this fibrin forms dense matted deposits. Although there are several factors that modify fibrin structure and delay fibrinolysis in these conditions, no mechanism is yet known to be responsible for a persistent presence of thrombi in the coronary and/or cerebral circulations. It seems, therefore, desirable to better understand this phenomenon in order to improve the effectiveness of thrombolytic therapies. Here, we show that ferric ions can activate non-enzymatic blood coagulation resulting in the formation of fibrin-like dense matted deposits (DMD) demonstrable by electron scanning microscopy (SEM). These DMDs are similar to those found in stroke and diabetes. On the basis of these findings we can conclude that the spontaneous formation of fibrin-like dense deposits in patients' blood may be a consequence of what is known as iron overload. Therefore, it is possible that inactivation of unbound iron in blood by small molecular weight chelating agents may prevent thrombotic consequences of the excessive accumulation of iron in the circulation.

Hydroxyl radical-modified fibrinogen as a marker of thrombosis: the role of iron.

Excessive free iron in blood and in organ tissues (so called iron overload) has been observed in degenerative diseases such as atherosclerosis, cancer, neurological, and certain autoimmune diseases, in which fibrin-like deposits are also found. Although most of the body iron is bound to hemoglobin and myoglobin in a divalent ferrous form, a certain amount of iron exists in blood as a trivalent (ferric) ion. This particular chemical state of iron has been shown to be toxic to the human body when not controlled by endogenous and/or dietary chelating agents. Experiments described in this paper show for the first time that ferric ions (Fe(3+)) can generate hydroxyl radicals without participation of any redox agent, thus making it a special case of the Fenton reaction. Ferric chloride was also demonstrated to induce aggregation of purified fibrinogen at the same molar concentrations that were used for the generation of hydroxyl radicals. Iron-aggregated fibrinogen, by contrast to native molecule, could not be dissociated into polypeptide subunit chains as shown in a polyacrylamide gel electrophoresis. The mechanism of this phenomenon is very likely based on hydroxyl radical-induced modification of fibrinogen tertiary structure with the formation of insoluble aggregates resistant to enzymatic and chemical degradations. Soluble modified fibrinogen species can be determined in blood of thrombotic patients by the reaction with protamine sulfate and/or by scanning electron microscopy. In view of these findings, it is postulated that iron-induced alterations in fibrinogen structure is involved in pathogenesis of certain degenerative diseases associated with iron overload and persistent thrombosis. It is concluded that the detection of hydroxyl radical-modified fibrinogen may be utilized as a marker of a thrombotic condition in human subjects.

Fast generation of stereolithographic models.

In this paper we present a work-in-progress method for fast and efficient generation of stereolithographic models. The overall approach is embedded in our general software framework Julius, which runs on high-end-graphic systems as well as on low-level PCs. The design of the support structures needed for the stereolithographic process will allow semiautomatic generation of the model. We did produce support structures for stereolithographic models with this fast data processing pipeline and will show future perspectives in this paper.

Julius--a software framework for computer-aided-surgery.

In the paper we introduce Julius--an extendable cross-platform software framework for medical visualization and surgical planning. Julius features a modular, cross-platform design using Qt and Vtk libraries and comes with a set of image analysis components, like semi-automatic segmentation, registration, visualization and navigation. We also present a 3D surface generation pipeline used in Julius for generating surfaces from volume data. The pipeline consists of image based filtering, marching cubes surface extraction algorithm, surface decimation and surface smoothing steps. We use this approach within different medical applications like craniofacial surgical planning and will also show the overall software framework within the paper.

Pathophysiology of oxidative stress in diabetes mellitus.

Oxidative stress is believed to play an important role, albeit not fully recognized, in the development of vascular complications in diabetes mellitus (DM) particularly type 2. In the majority of studies, attention was focused on lipid oxidation, specifically on that of low-density lipoproteins (LDLs). More recent investigations have revealed that it is not only the lipid but also the apolipoprotein moiety of LDL that becomes oxidatively modified resulting in the formation of insoluble aggregates. Consequently, it has been documented that LDL aggregation was due to the hydroxyl radical-induced dityrosine crosslinking between apo B monomers. In DM patients with atherosclerotic complications, intravascular fibrous deposits were shown to contain, in addition to oxidized LDL, a fibrin-like material (FLM). This material is immunologically identical to fibrin that is normally formed as a result of intravascular activation of the blood coagulation cascade. Although DM patients with vascular disease display increased concentration of plasma fibrinogen (Fbg), the precursor of fibrin, no markers of full blown activation of blood coagulation could be found.

Resistance of cancer cells to immune recognition and killing.

It is well recognized that, in order for a wound to heal, the fibrin clot must be eliminated by fibrinolytic enzymes. In certain instances, however, fibrin is ineffectively degraded or even not degraded. For example, in pregnancy, the placenta contains a layer of fibrin (Nitabuck's layer) which presents as 'self' to the immune system. Similar situations have been observed in many solid tumors. A hypothesis is presented according to which tumor cells can escape detection and attack by the immune system in most cancer patients. The tumor dons a 'coat' of the host's own protein on its cell surface. The coat is composed of fibrin and of a polymeric form of human serum albumin (HSA) which, by contrast to pure fibrin, is resistant to fibrinolytic degradation. Such a coated tumor appears as 'self' to the immune system, and thus is not detected as a tumor by the immune system (i.e. natural killer cells). When tumors are prepared for in vitro assays against drugs, they are routinely treated with proteolytic enzymes (e.g. pepsin, or chymotrypsin, etc.) which dissolve the protein coat, exposing the tumor cell surface to the drug. Thus, the in vivo existence of a coat on the tumor surface may explain why some drugs have little or no effect in vivo, while the same drugs are active in vitro.

Reduced glutamate in the anterior cingulate cortex in depression: an in vivo proton magnetic resonance spectroscopy study.

BACKGROUND: Functional imaging studies suggest a specific role of the anterior brain regions in the pathogenesis of major depression. The aim of this study was to evaluate possible neurochemical alterations in the frontomesial cortex in patients with major depressive episode using in vivo proton magnetic resonance spectroscopy ((1)H-MRS). METHODS: Single voxel (1)H-MRS was performed in 19 patients with major depressive episodes and 18 age-matched healthy controls within the anterior cingulate cortex and the parietal white matter. Absolute concentrations were estimated for N-acetyl-aspartate, choline-containing compounds, total creatine, myo-inositol, unresolved glutamate and glutamine (Glx) and glutamate alone (Glu). Voxel composition was analyzed by image segmentation into cerebrospinal fluid (CSF), grey and white matter. RESULTS: MANOVA test for Glx and Glu using age, percent CSF and percent grey matter contribution as covariates yielded a significant group effect within the anterior cingulate due to decrease of Glx in patients (-10.4%, p =.013). Considering only severely depressed patients, both Glx and Glu (-14.3%, p =.03) showed a significant decrease. There was no significant group effect for the neuronal marker NAA, creatine, choline or myo-inositol in either localization. CONCLUSIONS: This study suggests a possible role of altered glutamatergic neurotransmission within the anterior cingulate in the pathogenesis of mood disorders. The otherwise unremarkable findings of major brain metabolites confirms lack of neurodegenerative or membrane metabolic changes in major depression.

Effect of magnesium on fibrin formation from lower molecular weight (LMW) fibrinogen.

Fibrinogen circulating in human blood is comprised of high molecular weight (HMW) and lower molecular weight (LMW) fractions. As previously documented by means of SDS-polyacrylamide gel electrophoresis (PAGE), LMW fraction was significantly increased in patients with cardiovascular disease and with diabetes mellitus (DM). We have recently observed that the values of fibrinogen measured by thrombin clotting time (the method of Clauss) were consistently lower in EDTA plasma than those obtained with citrated plasma. However, supplementation of EDTA plasma with magnesium (Mg) ions gave comparable results. In this study we documented by SDS-PAGE that fibrin formed with thrombin alone in EDTA plasma originated from HMW fibrinogen, whereas that formed after addition of Mg was derived from LMW fibrinogen. Thus, measurement of thrombin clotting time in EDTA plasma with and without Mg may serve as a quick method for the determination of HMW and LMW fibrinogens in human blood. Preliminary result obtained with this new method revealed that LMW fibrinogen was significantly increased in DM patients. We have therefore concluded that measurement of this fraction of fibrinogen may prove to be of clinical diagnostic significance.

Signal undershoots following visual stimulation: a comparison of gradient and spin-echo BOLD sequences.

Gradient-echo (GRE) and spin-echo (SE) EPI BOLD sequences were used to quantitate the effect of visual stimulation. Both sequences showed a positive BOLD response during stimulation and a negative BOLD response in the interstimulation intervals. The relaxation rate changes during stimulation were larger for the GRE sequence than for the SE sequence, whereas in the interstimulation intervals they were not significantly different. In both cases, the ratio of the GRE/SE relaxation rate changes were consistent with BOLD effects in larger vessels despite the well-known lower sensitivity of the SE sequence to the extravascular component of the BOLD effect in larger vessels. The most probable explanation of this result is that a significant fraction of the observed changes originated from the intravascular component of the BOLD effect. The SE sequence depicted smaller areas of activation than the GRE sequence with more than 85% of the pixels being depicted as significant by the SE sequence being also significant in the GRE activation maps. However, for the reverse comparison, an overlap of only 35% was observed, with many of the strongly correlated GRE pixels showing weak correlations in the corresponding SE activation image. Our results, together with the fact that signal undershoots have not been observed by groups using MR sequences that measure absolute flow changes for similar stimulation paradigms, suggest that the undershoot may be due to alterations in the blood volume and/or hematocrit during stimulation that normalize at a slower rate than the changes in blood flow after the cessation of the stimulation, leading to a poststimulation signal undershoot.

Expectation maximization reconstruction of positron emission tomography images using anatomical magnetic resonance information.

Using statistical methods the reconstruction of positron emission tomography (PET) images can be improved by high-resolution anatomical information obtained from magnetic resonance (MR) images. We implemented two approaches that utilize MR data for PET reconstruction. The anatomical MR information is modeled as a priori distribution of the PET image and combined with the distribution of the measured PET data to generate the a posteriori function from which the expectation maximization (EM)-type algorithm with a maximum a posteriori (MAP) estimator is derived. One algorithm (Markov-GEM) uses a Gibbs function to model interactions between neighboring pixels within the anatomical regions. The other (Gauss-EM) applies a Gauss function with the same mean for all pixels in a given anatomical region. A basic assumption of these methods is that the radioactivity is homogeneously distributed inside anatomical regions. Simulated and phantom data are investigated under the following aspects: count density, object size, missing anatomical information, and misregistration of the anatomical information. Compared with the maximum likelihood-expectation maximization (ML-EM) algorithm the results of both algorithms show a large reduction of noise with a better delineation of borders. Of the two algorithms tested, the Gauss-EM method is superior in noise reduction (up to 50%). Regarding incorrect a priori information the Gauss-EM algorithm is very sensitive, whereas the Markov-GEM algorithm proved to be stable with a small change of recovery coefficients between 0.5 and 3%.

In vitro preparation and partial characterization of a novel complex of human fibrinogen with albumin.

Limited reduction of a mixture of purified human fibrinogen and human serum albumin leads to the formation of a stable complex. This complex is clottable with thrombin, but its electrophoretic mobility is lower than that of fibrinogen. By contrast to clots prepared from purified fibrinogen, those obtained from the complex are transparent and are completely resistant to fibrinolytic degradation induced with urokinase. In this respect the complex of fibrinogen with albumin is similar to that identified in congenitally abnormal fibrinogens.