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SummaryNoninvasive diagnosis of deep vein thrombosis (DVT) is based on ultrasound examination of the leg veins, usually restricted to only compression of the proximal veins (CUS). Patients with negative CUS findings require a second examination or a combination with other tests, which impairs clinical efficiency. In this prospective outcome study, 1646 consecutive patients with clinically suspected DVT were examined once by a standardized protocol of complete compression ultrasound comprising all proximal and distal veins (CCUS) as the only diagnostic test. The examination was equivocal in 15 patients (1% technical failure rate). Another 366 patients (22%) were tested positive for proximal DVT, distal DVT, muscle vein thrombosis, or phlebitis. Of 1265 patients in whom CCUS findings were negative, 242 met exclusion criteria for follow-up (age

The thrombin activatable fibrinolysis inhibitor (TAFI) influences the pathways regulating fibrin formation and deposition. The enormous TAFI plasma level variability present in adults may be explained by a combination of two polymorphisms in the TAFI gene (+1542C>G; 505G>A). We aimed to correlate these two polymorphisms with plasma TAFI antigen concentrations in healthy children and pediatric oncology patients with and without venous thrombosis who were supplied with Broviac central venous catheters. Polymorphisms were detected by restriction fragment length polymorphism (RFLP) analysis of polymerase chain reaction (PCR) amplification, whereas TAFI concentration was determined using a commercial enzyme-linked immunosorbent assay (ELISA). Samples from 57 controls and 67 pediatric patients (11 venous thrombotic complications) were studied. TAFI levels in healthy children and patients were not influenced by gender or age. Compared with the 505GG carriers (wild type), 505AA carriers as well as heterozygous 505GA carriers each exhibited significantly higher TAFI antigen concentrations. In contrast, the lowest TAFI levels were detected in homozygous carriers of the +1542GG polymorphism. A combination of the genotype 505AA (homozygous carrier) and +1542CC (wild type) was present in 13 probands and resulted in the highest TAFI levels. Although in oncologic patients the risk of thrombosis was markedly increased by the heterozygous factor V 1691G>A mutation, the two TAFI polymorphisms investigated exerted no thrombogenic influence.

Factor V Leiden is a well-known risk factor for venous thrombosis. The dual role of factor V as a coagulatory and anticoagulatory cofactor permits the assumption that further mutations in the factor V gene are of importance in the study of the risk of thrombosis. Relevant studies to date have given rise to a controversy over this risk for the HR2 haplotype. For the G allele, defined in our work group as a G at the nucleotide positions 2391, 2663, 2684 and 2863, there have been to date no other investigations of thrombotic risk. In a case-control study on 347 patients with deep venous thrombosis (DVT) and 282 controls, we investigated the association of the HR2 haplotype and the G allele with DVT. We found no association between HR2 haplotype and DVT [odds ratio (OR) 0.87; 95% confidence interval (CI) 0.58-1.30; P = 0.537]. The frequency of the G allele was, on the contrary, higher in the control group than among the patients (OR 0.68, 95% CI: 0.53 to 0.89; P = 0.005). The factor V activity of the HR2 carriers was lower than that of the wild type and G allele carriers. The HR2 haplotype exhibited a moderate influence on activated protein C response. This study presented no evidence of thrombotic risk for the HR2 haplotype alone. The results here permit the assumption of a protective effect of the G allele. The source of a possible protective influence of the G allele on thrombotic risk is at present unclear.

Silibinin, a naturally-occurring flavonoid produced by milk thistle, possesses antioxidant, anti-inflammatory and cancer-preventive activities. In the current study, we examined the effects of silibinin on the expression of secreted phospholipase A2 (sPLA2) enzymes, especially those of group IIA (hGIIA), which play a crucial role in inflammation and carcinogenesis.The effects of silibinin on sPLA2 expressions in human HepG2 hepatoma and PC-3 prostate cancer cells were analyzed using quantitative reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay technique.Silibinin inhibited the expression of hGIIA in unstimulated and cytokine-primed HepG2 and PC-3 cells. The mRNA levels of sPLA2 of groups IB, III and V were also significantly decreased by silibinin. Analyses of transcription factor activation suggest that nuclear factor-κB, but not specificity protein 1 (SP1) is implicated in the silibinin-mediated down-regulation of hGIIA.Silibinin exhibits inhibitory effects on basal and cytokine-induced expression of sPLA2s in cancer cells and therefore, may have the potential to protect against up-regulation of hGIIA and other sPLA2 isoforms during inflammation and cancer.

Background: Comparisons of aldosterone with cortisol during adrenal venous sampling (AVS) provide the currently recommended technique for distinguishing unilateral adenomas from bilateral adrenal hyperplasia during the diagnostic work-up of patients with primary aldosteronism. Selectivity of AVS is routinely established from adrenal venous (AV) to peripheral venous (PV) ratios of cortisol, but is more accurately indicated by AV: PV ratios of plasma metanephrine. Objective: Here we describe a novel liquid-chromatography tandem mass spectrometric (LC-MS/MS) assay for profiling plasma levels of 15 adrenal steroids during AVS, with and without ACTH stimulation. Methods: Selectivity of AVS was established by AV: PV ratios of plasma metanephrine larger than 12. Concentrations of aldosterone, corticosterone, 11-deoxycorticosterone, progesterone, pregnenolone, cortisone, cortisol, 11-deoxycortisol, 17-hydroxy-progesterone, androstenedione, dehydroepiandrosterone, dehydroepiandrosterone-sulfate, 21-deoxycortisol, 18-oxo-cortisol and 18-hydroxy-cortisol were compared in PV and AV plasma samples from 35 patients with and without ACTH stimulation. Aldosterone and cortisol levels measured by LC-MS/MS were compared with levels measured by immunoassay. Results: Median AV: PV ratios of plasma cortisol (17) were lower than those of corticosterone (52), 11-deoxycorticosterone (68), pregnenolone (72), 11-deoxycortisol (55) and 17-hydroxy-progesterone (57). ACTH stimulation increased AV plasma concentrations of cortisol 6-fold, but had larger effects on other steroids such as corticosterone, which increased by 36-fold. As a result AV plasma concentrations of corticosterone relative to cortisol increased from 6% to 38%. Method comparisons indicated agreement of concentrations of cortisol measured by immunoassay and LC-MS/MS. Conclusion: Compared to other adrenal steroids, cortisol provides a less sensitive marker for selectivity of AVS.

Research generally supported a positive effect of competition intensity on financial commitment into news production within U.S. markets. The authors test this assumption across European newspaper markets by applying fuzzy set qualitative comparative analysis. Although oligopolistic too, European newspaper markets reveal a somewhat different pattern. The market condition “high number of competitors” in combination with “high competition intensity” is a sufficient condition for financial commitment across the different markets in this study.

The courses of 79 children (2 weeks to 19 years old) treated with two different low-molecular weight heparins (LMWHs)--nadroparin (n=66) and enoxaparin (n=13)--were retrospectively analysed. In 62 patients, LMWHs were given for short-term prophylaxis (1-2 weeks) during immobilization after surgery or trauma. Thirteen children with thromboembolic events received long-term prophylaxis with LMWHs for 2-18 months--six after thrombolytic therapy and seven after therapy with unfractionated heparin (UFH). Because of thromboembolic events, four patients were initially treated with LMWHs. In all patients with short-term prophylaxis, no thrombosis occurred. After thrombolytic therapy, three children had no reocclusion, two had no thrombus apposition and one had complete recanalization. In the seven patients treated with LMWHs after UFH, four had no reocclusion, two had recanalization and one had reocclusion. In all patients receiving LMWHs for initial treatment of thrombosis, no thrombus apposition, but also no recanalization, occurred. For short-term prophylaxis, nadroparin was used independent of the body weight and without determination of anti-factor Xa (anti-FXa) activity. Long-term prophylaxis was given mainly as doses of 45-100 anti-FXa U/kg resulting in anti-FXa activities between 0.2 and 0.4 U/ml. For treatment of thrombosis, doses of 200-300 anti-FXa U/kg corresponded to 0.5-1.0 anti-FXa U/ml. Side effects--slight gastrointestinal bleeding and temporary reversible hair loss--were seen in two patients. In conclusion, LMWHs proved to be efficacious and safe especially in prophylaxis of thromboembolic events in children.

Essential thrombocythaemia in a teenage girl resulting in acquired von Willebrand syndrome with joint haemorrhage and menorrhagia

So far little is known about the importance of different types of non-diabetic hyperglycemia for the development of macrovascular disease. The aim of this work was to examine the intima-media thickness (IMT) of the common carotid artery (CCA), a well-accepted marker of atherosclerosis, as well as various risk factors for atherosclerosis in non-diabetic subjects with isolated fasting (IFH; n=67), isolated postchallenge (IPH; n=82) and combined hyperglycemia (CH; n=88) in comparison to normoglycemic (NG; n=265) controls. Subjects were participants of the RIAD study (Risk Factors in IGT for Atherosclerosis and Diabetes). IMT in the IPH (IMTmean: 0.89+/-0.02 mm; IMTmax: 1.01+/-0.02 mm; mean+/-SEM) and CH group (IMTmean: 0.91+/-0.02 mm; IMTmax: 1.03+/-0.02 mm) was significantly increased vs. the NG (IMTmean: 0.82+/-0.01 mm; IMTmax: 0.94+/-0.01 mm) and IFH group (IMTmean: 0.81+/-0.02 mm; IMTmax: 0.90+/-0.03 mm). IMT of the IFH group was similar to the normoglycemic controls. Subjects in the first and second tertile for postchallenge plasma glucose have similar carotid IMT irrespective of the level of fasting plasma glucose. The individuals of the third tertile for 2 h plasma glucose, whether in the first, second or third tertile of fasting plasma glucose, showed the same carotid IMT, which was significantly higher than all other groups, except for the one with lowest tertile for fasting and postchallenge plasma glucose. Except for total cholesterol and von Willebrand factor the levels of all other risk parameters were significantly higher in the hyperglycemic groups in comparison to the normoglycemic controls. Among the hyperglycemic subjects the CH group was at the highest risk for atherosclerosis with significantly increased levels of plasma triglycerides, fibrinogen, PAI-1, albuminuria, HDL-triglycerides, free fatty acids, insulin and proinsulin, and significantly reduced HDL-cholesterol in comparison to the normoglycemic controls. In summary, postchallenge hyperglycemia within the non-diabetic range is associated with atherosclerosis, as measured by the increased intima-media thickness of the common carotid artery. Furthermore, cardiovascular risk factors are significantly raised in all types of non-diabetic hyperglycemia.

The factor V (FV) B-domain is extremely important to the cofactor function of native FV for activated protein C (APC) in the inactivation of factor VIII (FVIII). In a previous study, we found that in the B-domain coding portion of DNA, the polymorphisms at nucleotide positions 2391, 2663, 2684, and 2863 were associated. In the major allele, all bases are A (A allele) and those in the minor allele are G (G allele). This study concerns itself with the question of whether or not there are differences in the APC response between the A allele and the G allele in plasma samples from persons without the FV Leiden. The APC ratios of homozygous carriers of the major A allele and the minor G allele do not differentiate themselves in classical activated partial thromboplastin time-based assays. In contrast, a test based on the deactivation of FVIII in the tenase complex in homozygous carriers of the minor G allele showed significantly lower APC ratios (P = 0.001) in comparison with the major A allele. The results of the investigation after modification of the test indicate that mutative changes in the B-domain apparently influence the interaction among phospholipids, APC, FV, and protein S. An increase in FVIII through the introduction of the FVIII concentrate Kogenate to the plasma samples was associated with a drop in the APC ratios of both genotypes. After defining 59 age- and sex-based matched pairs without the FV Leiden, the observed frequency of the minor G allele was higher in the non-thrombotic group (33.0%) than in the thrombotic group (22.8%). However, the difference did not reach the level of significance (odds ratio, 0.53; 95% confidence interval, 0.26-1.12). It does, nevertheless, appear possible that a homozygous condition for the minor allele in combination with a defect known to be associated with thrombophilia represents an additional thrombogenetic risk factor.

To evaluate the role of inherited thrombophilia in the development of central venous line (CVL)-related thrombosis, the following parameters were determined in 77 pediatric-oncologic patients with CVL: activated protein C (APC)-ratio, factor V (FV) G1691A and prothrombin G20210A mutation, protein C, protein S, antithrombin, coagulation factor XII, lipoprotein (a) and homocysteine. An inherited prothrombotic risk factor was found in 17 patients (23%). Four out of 14 patients with a single defect (hyperlipoproteinemia, heterozygous FV G1691A and prothrombin G20210A mutation, protein C deficiency type I) and all three patients with combined defects (heterozygous FV G1691A mutation combined with heterozygous prothrombin G20210A variant, protein S deficiency or hyperlipoproteinemia) suffered from CVL-related thrombosis. In 11 out of 77 patients (14%) a CVL-related thrombosis was detected. In 2 children thrombosis occurred a few days after asparaginase therapy and in another three thrombosis was associated with CVL-related septicemia caused by Staphylococcus epidermidis. After removal of CVL, thrombosis was detected in 5 children, in 2 without clinical symptoms but in the presence of inherited prothrombotic risk factors. Conclusion The present study demonstrates the clinical importance of CVL in combination with inherited thrombophilia in the development of thrombosis in pediatric-oncologic patients. Before or shortly after insertion of CVL, patients should be tested for the presence of factor V G1691A mutation, prothrombin G20210A variant and increased lipoprotein (a) values.

To determine a new category of dysfunctional glucose homeostasis - impaired fasting glucose (IFG) - introduced by the American Diabetes Association (ADA) and the World Health Organization (WHO) defining those with abnormal but nondiabetic fasting glucose values and with a possible risk for developing diabetes. It is not known whether IFG is a risk factor for atherosclerosis, as is impaired glucose tolerance (IGT).In this case-control cross-sectional study in which the oral glucose tolerance (75-g OGTT) and the carotid intima-media thickness (IMT) with B mode ultrasound, as a marker of atherosclerosis, were measured, together with HbA1c, lipids, plasminogen activator (PAI), insulin and proinsulin concentrations in blood plasma. Out of 788 subjects of the risk factors in IGT for Atherosclerosis and Diabetes (RIAD) study we found 104 IFG cases that were compared to 104 controls with fasting plasma glucose (FPG)6.1 mmol/l, matched for age, sex and body mass index. Subjects with 2h postprandial (pp) plasma glucoseor = 11.1 mmol/l were excluded. The rest were subdivided into those with 2h plasma glucose7.8 mmol/l (63 pairs, NGT) and those with plasma glucose7.8 mmol/l and11.1 mmol/l (41 pairs, IGT).The case and control groups showed no significant differences in the major risk factors except for waist-to-hip ratio (WHR) which was higher in the IFG with NGT. IFG with NGT exhibited significantly higher levels of HbA1c, true insulin and proinsulin. In IFG with IGT, only HbA1c and proinsulin were significantly increased vs. controls. IMT was in the same range for cases and controls in both subgroups. However, IMT mean and IMTmax were significantly increased in IFG with IGT vs. IFG with NGT (0.95 mm vs. 0.80 mm and 1.10 mm vs. 0.90 mm). Cumulative distribution analysis of IMT illustrates that IMT in IFG with IGT is more shifted to higher artery wall thickness than in IFG with NGT.In our case-control study IFG alone was not related to increased IMT. Only IFG in a combination with IGT exhibited atherosclerotic changes of the carotid arteries. IFG is not analogous to IGT as a risk factor for atherosclerosis.

ZusammenfassungZiel der Untersuchung war es, das Vorkommen von pathologischer APCResistenz und Faktor-V-Leiden-Mutation bei Patienten mit durchgemachter Phlebothrombose und peripherer arterieller Verschlußkrankheit aus unserem Krankengut zu ermitteln. Als Vergleichsgruppe dienten gefäßgesunde Probanden, die unterteilt wurden in Probanden mit positiver Thrombose-Familienanamnese und Probanden mit thrombosefreier Familienanamnese. In die Untersuchung einbezogen wurden 132 Probanden (64 Frauen und 68 Männer, mittleres Alter 48,5 ± 15,5 Jahre).Der relative Anteil pathologischer APC-Response betrug in der Gruppe der Patienten mit tiefer Beinvenenthrombose 42% und in der Gruppe der Patienten mit peripherer arterieller Verschlußkrankheit 32%. In beiden Patientengruppen konnte die gestörte APC-Response nur bei ca. 50% durch eine Faktor-V-Leiden-Mutation erklärt werden. Bei gefäßgesunden Probanden war eine pathologische APC-Ratio dagegen fast ausschließlich durch die Faktor-V-Leiden-Mutation bedingt. Der relative Anteil der pathologischen APC-Response betrug bei Probanden ohne Thromboseanamnese in der Familie 12,5%, mit positiver Familienanamnese 41%. Der hohe Anteil gefäßgesunder Probanden mit positiver Familienanamnese unterstreicht das moderate Thromboserisiko bei Faktor-V-Leiden-Mutation ohne zusätzliche Risikofaktoren. Die Ursache der pathologischen APC-Response bei Patienten ohne Faktor-V-Leiden-Mutation ist bisher unklar.Eine pathologische APC-Ratio bei Patienten mit einem Wildtyp für Faktor-V-Leiden sollte Anlaß für eine Verlaufskontrolle möglichst unter Einbeziehung weiterer Familienmitglieder sein, um Hinweise auf andere genetische oder funktionelle Ursachen für eine pathologische APC-Response zu bekommen.

ZusammenfassungBei 157 Patienten und 14 Familienangehörigen wurde die Response gegenüber aktiviertem Protein C (APC) in drei unterschiedlichen Tests bestimmt und mit dem Ergebnis der Genotypisierung verglichen. Die Ermittlung der laboreigenen Referenzwerte erfolgte aus Plasmaproben von 96 anamnestisch gesunden Probanden mit normalem Genotyp. Zur Bestimmung der APC-Response wurden folgende Testsysteme eingesetzt: Test Immunochrom® APC-Response, Coatest® APC™ Resistance und Coatest® APC™ Resistance mit einer Verdünnung des Plasmas mit Faktor-V-Mangelplasma. Die Testmodifikation mit Verdünnung des Patientenplasmas mit Faktor-V-Mangelplasma erbrachte mit Ausnahme von Patienten unter Vollheparinisierung eine gute Trennung zwischen Wildtyp und Faktor-V-Mutation. Dieser Test reduziert jedoch offensichtlich die Aussage auf die Faktor-V-Mutation. Der Test Immunochrom® APC-Response zeigte unabhängig von einer Antikoagulation eine gute Übereinstimmung mit dem Ergebnis der Genotypisierung. Obwohl ein großer Teil der APC-Resistenz assoziiert ist mit einer Mutation im Faktor-V-Gen (Leiden-Mutation) sind beide Begriffe nicht synonym. Pathologische Ratios bei Patienten mit einem Wildtyp unterstreichen die Notwendigkeit zur Bestimmung der funktionellen Response und des Genotypes, ihre Ursache bedarf weiterer Abklärung.

Background Medication-related interferences with measurements of catecholamines and their metabolites represent important causes of false-positive results during diagnosis of phaeochromocytomas and paragangliomas (PPGLs). Such interferences are less troublesome with measurements by liquid chromatography with tandem mass-spectrometry (LC-MS/MS) than by other methods, but can still present problems for some drugs. Levodopa, the precursor for dopamine used in the treatment of Parkinson’s disease, represents one potentially interfering medication. Methods Plasma and urine samples, obtained from 20 Parkinsonian patients receiving levodopa, were analysed for concentrations of catecholamines and their O-methylated metabolites by LC-MS/MS. Results were compared with those from a group of 120 age-matched subjects and 18 patients with PPGLs. Results Plasma and urinary free and deconjugated (free + conjugated) methoxytyramine, as well as urinary dopamine, showed 22- to 148-fold higher ( P Conclusions These data establish that although levodopa treatment confounds identification of PPGLs that produce dopamine, the therapy is not a problem for use of LC-MS/MS measurements of plasma and urinary normetanephrine and metanephrine to diagnose more commonly encountered PPGLs that produce noradrenaline or adrenaline.

Arthrocentesis is an essential emergency step in managing patients with acute arthritis. To identify a bacterial infection, Gram staining is performed promptly. However, crystal analysis may not be immediately performed in many facilities. Being considered not to be stable over time, synovial fluid (SF) is sometimes discarded instead of being stored for crystal identification.The aim of this study was to assess the detectability of monosodium urate (MSU) and calcium pyrophosphate (CPP) crystals in SF over a period of 3 days.Consecutive SF samples from 75 joints were analyzed for MSU, CPP crystals, and pH. Two independent observers evaluated the samples by regular light and polarization microscopy immediately after arthrocentesis and after 1, 2, and 3 days at room temperature or at 4°C.Of 75 samples, 27 contained crystals (16 MSU, 6 CPP, 5 both); semiquantitative counts of both MSU and CPP crystals did not change significantly after 3 days. There was no new formation of crystals in any of the crystal-negative samples, which was independent of the storage temperature. Synovial fluid pH was not predictive of crystals and did not change over time.Although immediate workup for microbiology, including Gram stain and culture, is indispensable and well established, crystal analysis may at times not be immediately performed. Our study suggests that when crystal identification cannot be done immediately, it can be safely performed up to 3 days after arthrocentesis when SF is stored at 4°C or even at stable room temperature (20°C).

Serum amyloid A (SAA), secreted group IIA phospholipase A₂ (sPLA₂-IIA), and C-reactive protein (CRP) are acute-phase proteins whose serum concentrations increase not only during inflammatory disorders, but also in the course of malignant diseases.In this study we analyzed serum levels of these inflammatory markers along with prostate-specific antigens (PSA) in patients with benign prostatic hyperplasia (BPH, n = 55), localized prostate cancers (PCa, n = 55), and metastatic prostate cancers (mPCa, n = 27) using immunological assays.We found that in comparison to healthy individuals (n = 55), patients with BPH, PCa and mPCa have elevated serum levels of SAA, sPLA₂-IIA, and CRP, in addition to elevated levels of PSA. Significant differences with respect to inflammatory biomarkers were found between localized and metastatic PCa (p0.001), suggesting a prognostic value of these parameters. In addition, serum concentrations of SAA and sPLA₂-IIA positively correlate with CRP in BPH patients (p0.05) and in patients with PCa and mPCa (p0.001), but not with PSA levels, Gleason score, or tumor stage, emphasizing a role of SAA and sPLA₂-IIA as circulating biomarkers of inflammation rather than of neoplastic transformation. In contrast to PSA, which differed significantly between BPH and localized PCa patients (p0.01), such a difference was not found for SAA, sPLA₂-IIA, and CRP. In order to elucidate whether the elevated levels of SAA and sPLA₂-IIA can be caused by cancer cell-associated synthesis, in vitro studies were performed. These analyses demonstrated the expression of SAA and sPLA₂-IIA in LNCaP and PC-3 prostate cell lines, which can be further upregulated by pro-inflammatory cytokines in a cell type-dependent manner. This might suggest that, in addition to the hepatic origin, SAA and sPLA₂-IIA can also be synthesized and secreted by prostatic cancer tissue itself.The results of the present study emphasize the utility of SAA, sPLA₂-IIA, and CRP as circulating biomarkers of inflammation during BPH development and PCa progression.

Background Measurements of plasma normetanephrine (NMN) and metanephrine (MN) provide a sensitive test for diagnosis of phaeochromocytomas and paragangliomas (PPGLs), but do not allow detection of dopamine-producing tumours. Here we introduce a novel mass spectrometric based method coupled to ultraperformance liquid chromatography (LC-MS/MS) for measuring NMN, MN and 3-methoxytyramine (MTY), the O-methylated metabolite of dopamine. Methods Specific collision-induced fragment ions assessed by multireaction monitoring transitions were used for identification, with quantification according to signal intensities of analytes relative to stable isotope labelled internal standards. Results for solid-phase extracted samples from 196 subjects analysed by LC-MS/MS were compared with those analysed by liquid chromatography with electrochemical detection (LC-ECD). Concentration ranges in 125 volunteers were compared with those from 63 patients with PPGLs, including 14 with metastatic disease. Results The LC-MS/MS method showed linearity over four orders of magnitude with analytical sensitivity sufficient to measure to 0.02 nmol/L. Intra- and inter-assay coefficients of variation ranged from 2.8% to 13.5%. NMN and MN were respectively measured 17% and 10% higher and MTY 26% lower by LC-MS/MS than by LC-ECD. Medians and ranges for 3-methoxytramine, NMN and MN were respectively 0.08 (0.03-0.13), 0.35 (0.13-0.95) and 0.15 (0.07-0.33) nmol/L in volunteers. Among patients with PPGLs, plasma methoxytyramine was six-fold higher in patients with than without metastastases (1.09 versus 0.19 nmol/L) and in three patients was the only metabolite increased. Conclusions The LC-MS/MS method enables accurate, selective and sensitive measurements of catecholamine O-methylated metabolites that should be particularly useful for screening and management of dopamine-producing metastatic PPGLs.

There is evidence of iron deficiency (ID) in patients treated with lipoprotein apheresis. Aim of this study was to assess ID in apheresis patients and to study its management comparing safety and efficacy of two approved intravenous (i.v.) iron formulations.Inclusion criteria were defined as a) serum ferritin300 μg/l and transferrin saturation20%, b) ferritin100 μg/l. Both iron deficient alone and ID anemic (IDA) patients were included. Other causes for anemia were ruled out by thorough history-taking and examination/blood tests. Patients were treated with six different lipoprotein apheresis methods: DALI, Liposorber D, TheraSorb LDL, HELP, MONET and Lipidfiltration. 50 patients were randomized to either ferric carboxymaltose (FCM, 500-1000 mg as single shot infusion over 20 min) or ferric gluconate (FG, 62.5 mg once weekly).50 of 67 patients of our Lipoprotein Apheresis Center showed iron deficiency. Both i.v. iron formulations studied were equally safe (no serious adverse events (SAEs), 6 patients/group showed adverse events (AEs)) and both effective (clinically and with respect to laboratory data) in lipoprotein apheresis patients, however FCM led to a more rapid and steeper rise of iron parameters.ID and IDA are common findings in lipoprotein apheresis patients. The pathogenesis remains yet poorly understood and is probably multifactorial. Differential diagnosis of ID/IDA is as essential as differential therapy. Handled with care, older i.v. iron preparations like FG appear to be safe and effective in lipoprotein apheresis patients. However, novel formulations like FCM can be administered rapidly at higher doses due to high complex stability, allowing faster filling of iron stores. Newer laboratory parameters (Reticulocyte-He, low/medium/high fluorescence reticulocytes (LFR/MFR/HFR)) assessing iron status may be helpful in early detection of ID and in monitoring iron replacement therapy.

Plasminogen activator inhibitor-1 (PAI-1) and urokinase-type plasminogen activator (uPA) play a crucial role in cancer progression. In the present study we examined the regulation of PAI-1 and uPA expressions in normal prostate epithelial cells (PrEC) and the prostate cancer cell lines LNCaP, DU-145, and PC-3. The antigen and mRNA levels of PAI-1 were down-regulated in cancer cells, especially in LNCaP and DU-145. In the presence of proinflammatory cytokines, an increase of PAI-1 mRNA levels was observed in PrEC, LNCaP and PC-3, but not in DU-145 cells. Treatment with demethylating agent, 5-aza-2'-deoxycytidine increased the level of PAI-1 transcript in DU-145 cells and restored the inducing effect of cytokines on PAI-1 expression. An aberrant methylation of PAI-1 promoter in DU-145 and LNCaP cells was shown by methylation-sensitive high resolution melting (MS-HRM) analysis. PAI-1 methylation was also significantly increased in tumor samples (23.2±1.7%) in comparison to adjacent non-tumor tissue (6.0±0.8%). Furthermore, the expression of uPA was increased in high invasive cell lines DU-145 and PC-3 in comparison to PrEC and low invasive LNCaP cells. MS-HRM analysis revealed aberrant methylation of uPA promoter in LNCaP cells, but not in PrEC, DU-145 and PC-3 cells, as well as in normal and prostate cancer tissue samples. In conclusion, the study shows that PAI-1 and uPA expressions were changed in opposite directions in high invasive prostate cancer cell lines resulting in a strong decrease of PAI-1/uPA ratio, which may indicate a shift towards proteolytic activities. Methylation of the PAI-1 gene is suggested as one of the molecular mechanisms involved in the cancer-associated down-regulation of the PAI-1 expression.

Background Measurements of plasma normetanephrine and metanephrine provide a useful diagnostic test for phaeochro-mocytoma, but this depends on appropriate reference intervals. Upper cut-offs set too high compromise diagnostic sensitivity, whereas set too low, false-positives are a problem. This study aimed to establish optimal reference intervals for plasma normetanephrine and metanephrine. Methods Blood samples were collected in the supine position from 1226 subjects, aged 5-84 y, including 116 children, 575 normotensive and hypertensive adults and 535 patients in whom phaeochromocytoma was ruled out. Reference intervals were examined according to age and gender. Various models were examined to optimize upper cut-offs according to estimates of diagnostic sensitivity and specificity in a separate validation group of 3888 patients tested for phaeochromocytoma, including 558 with confirmed disease. Results Plasma metanephrine, but not normetanephrine, was higher (P < 0.001) in men than in women, but reference intervals did not differ. Age showed a positive relationship (P < 0.0001) with plasma normetanephrine and a weaker relationship (P = 0.021) with metanephrine. Upper cut-offs of reference intervals for normetanephrine increased from 0.47 nmol/L in children to 1.05 nmol/L in subjects over 60 y. A curvilinear model for age-adjusted compared with fixed upper cut-offs for normetanephrine, together with a higher cut-off for metanephrine (0.45 versus 0.32 nmol/L), resulted in a substantial gain in diagnostic specificity from 88.3% to 96.0% with minimal loss in diagnostic sensitivity from 93.9% to 93.6%. Conclusions These data establish age-adjusted cut-offs of reference intervals for plasma normetanephrine and optimized cut-offs for metanephrine useful for minimizing false-positive results.

SummaryThe plasma tissue factor (TF) concentration was correlated to factor VII concentration (FVIIag) and factor VII activity (FVIIc) in 498 healthy volunteers ranging in age from 17 to 64 years. Immunoassays using monoclonal antibodies (mAbs) were developed for the determination of TF and FVIIag in plasma. The mAbs and the test systems were characterized. The mean value of the TF concentration was 172 ± 135 pg/ml. TF showed no age- and gender-related differences. For the total population, FVIIc, determined by a clotting test, was 110 ± 15% and the factor VIlag was 0.77 ± 0.19 μg/ml. FVII activity was significantly increased with age, whereas the concentration demonstrated no correlation to age in this population. FVII concentration is highly correlated with the activity as measured by clotting assay using rabbit thromboplastin. The ratio between FVIIc and FVIIag was not age-dependent, but demonstrated a significant difference between men and women. Between TF and FVII we could not detect a correlation.

Tissue injury following trauma and surgery may induce alterations in blood coagulation and fibrinolysis. Hypercoagulable state after surgery can be associated with the risk of postoperative thromboembolic complications. The contact of coagulation factors with TF after injury of vessel wall and organ tissues may contribute to the development of thrombosis after surgery (1). TF, the cell surface receptor and cofactor of factor VII/VIIa is normally not expressed by cells within the vasculature. Only monocytes and endothelial cells can be stimulated to express TF transiently by a variety of inflammatory and immunological reactions (for review see 2,3). Also surgical treatment was reported to induce TF synthesis in monocytes (4,5,6). TF is present in many extravascular tissues as vascular adventitia, organ capsules, epidermis, colonic mucosal epithelium, liver stroma, pancreas stroma and also on tumor cells (7-12). In this study, we investigated, whether we can detect the release of TF from the traumatized tissues and from activated monocytes into the circulation following abdominal surgery. To test the dependence of the extension of tissue injury during surgery we segregated the patients into group A with major abdominal operations and group B consisting of patients with appendectomy and cholecystectomy. No relationship could be established between changes of TF and postoperative thromboembolic complications.

BACKGROUND: We first describe a patient who developed urosepsis from an ordinary urinary tract infection. In this case, the new hematological parameters of immature leukocytes, that is, the high‐fluorescence lymphocyte cell (HFLC) and immature granulocyte (IG) counts peaked early, whereas the established infection parameters, that is, C‐reactive protein (CRP) and total white blood cell count showed less dynamic regarding infection and therapy. METHODS: To investigate this phenomenon in greater detail, the novel parameters HFLC and IG counts are investigated retrospectively in a cohort of 38 patients who were admitted to the anesthesia intensive care unit. Three groups of patients have been analyzed and compared: patients without signs of infection, patients with limited infections, and patients with sepsis. Data were collected with a Sysmex XE‐5000 hematological analyzer. RESULTS: In patients (n = 22) without any signs of infection, both values are very low. In patients with limited local infections (n = 10), moderate elevations of the IG and HFLC counts are seen. In patients with sepsis (n = 6), the IG and HFLC counts are significantly higher. CONCLUSION: The total IG count seems to be useful for distinguishing a septic patient from a nonseptic (P < 0.004). Hematological parameters have the advantage of being measured easily during routine blood cell analysis.

Objective We introduce a novel liquid chromatographic tandem-mass spectrometric method for simultaneous measurements of urinary catecholamines and their free O-methylated metabolites, which we compare to the deconjugated metabolites. Methods Method performance was validated for recovery, linearity, precision and accuracy, analyte stability, ion suppression and carry over. Results from 53 patients with and 138 volunteers without pheochromocytoma were compared. Results Analyte recoveries ranged from 60 to 96% and intra- and inter-assay coefficients of variation from 2.7 to 13.2%. The method showed excellent linearity over 3 orders of magnitude with analytical sensitivity sufficient to measure to 1.2 nmol/L. Free O-methylated metabolites were excreted at less than 20% the rates of the deconjugated metabolites, but were easily measureable. Increases in urinary normetanephrine in pheochromocytoma patients relative to volunteers were higher for free than deconjugated metabolites and higher for both than for norepinephrine (10 vs 5.5 vs 3.7 fold increases). In contrast, relative increases in urinary free versus deconjugated metanephrine (2.7 and 3.2) and methoxytyramine (2.1 and 1.9) did not differ, but for methoxytyramine were larger than for dopamine (1.2). Conclusion Measurements of urinary catecholamines and their free O-methylated metabolites by our method provide potential advantages over urinary deconjugated metanephrines for diagnosis of pheochromocytoma.

Background The M-type phospholipase A2 receptor (PLA2R1) plays a crucial role in several signaling pathways and may act as tumor-suppressor. This study examined the expression and methylation of the PLA2R1 gene in Jurkat and U937 leukemic cell lines and its methylation in patients with myelodysplastic syndrome (MDS) or acute leukemia. Methods Sites of methylation of the PLA2R1 locus were identified by sequencing bisulfite-modified DNA fragments. Methylation specific-high resolution melting (MS-HRM) analysis was then carried out to quantify PLA2R1 methylation at 5`-CpG sites identified with differences in methylation between healthy control subjects and leukemic patients using sequencing of bisulfite-modified genomic DNA. Results Expression of PLA2R1 was found to be completely down-regulated in Jurkat and U937 cells, accompanied by complete methylation of PLA2R1 promoter and down-stream regions; PLA2R1 was re-expressed after exposure of cells to 5-aza-2´-deoxycytidine. MS-HRM analysis of the PLA2R1 locus in patients with different types of leukemia indicated an average methylation of 28.9% ± 17.8%, compared to less than 9% in control subjects. In MDS patients the extent of PLA2R1 methylation significantly increased with disease risk. Furthermore, measurements of PLA2R1 methylation appeared useful for predicting responsiveness to the methyltransferase inhibitor, azacitidine, as a pre-emptive treatment to avoid hematological relapse in patients with high-risk MDS or acute myeloid leukemia. Conclusions The study shows for the first time that PLA2R1 gene sequences are a target of hypermethylation in leukemia, which may have pathophysiological relevance for disease evolution in MDS and leukemogenesis.

Here, we describe a novel method utilizing double stable isotope ultra performance liquid chromatography-tandem mass spectrometry to measure tissue contents and activity of phenylethanolamine N-methyltransferase (PNMT), the enzyme responsible for synthesis of the stress hormone, epinephrine. The method is based on measurement of deuterium-labeled epinephrine produced from the reaction of norepinephrine with deuterium-labeled S-adenosyl-l-methionine as the methyl donor. In addition to enzyme activity, the method allows for determination of tissue contents of PNMT using human recombinant enzyme for calibration. The calibration curve for epinephrine was linear over the range of 0.1 to 5,000 pM, with 0.5 pM epinephrine representing the lower limit of quantification. The calibration curve relating PNMT to production of deuterium-labeled epinephrine was also linear from 0.01 to 100 ng PNMT. Intra- and inter-assay coefficients of variation were respectively 12.8 % (n = 10) and 10.9 to 13.6 % (n = 10). We established utility of the method by showing induction of the enzyme by dexamethasone in mouse pheochromocytoma cells and strong relationships to PNMT gene expression and tissue epinephrine levels in human pheochromocytomas. Development of this assay provides new possibilities for investigations focusing on regulation of PNMT, the crucial final enzyme responsible for synthesis of epinephrine, the primary fight-or-flight stress hormone.

Die Berichterstattung uber politische Kampagnen in der Schweiz findet in einem Umfeld, in dem sich die Frage nach Pluralismus oder Populismus direkt stellt. Ein Umfeld, in welchem die Medien dem Druck politischer Akteure und PR-Berater ausgesetzt sind und sich Politiker standig prasentieren und rechtfertigen mussen, wofur die Medien ein wichtiges Mittel sind.

Item does not contain fulltext BACKGROUND: There are currently no reliable biomarkers for malignant pheochromocytomas and paragangliomas (PPGLs). This study examined whether measurements of catecholamines and their metabolites might offer utility for this purpose. METHODS: Subjects included 365 patients with PPGLs, including 105 with metastases, and a reference population of 846 without the tumour. Eighteen catecholamine-related analytes were examined in relation to tumour location, size and mutations of succinate dehydrogenase subunit B (SDHB). RESULTS: Receiver-operating characteristic curves indicated that plasma methoxytyramine, the O-methylated metabolite of dopamine, provided the most accurate biomarker for discriminating patients with and without metastases. Plasma methoxytyramine was 4.7-fold higher in patients with than without metastases, a difference independent of tumour burden and the associated 1.6- to 1.8-fold higher concentrations of norepinephrine and normetanephrine. Increased plasma methoxytyramine was associated with SDHB mutations and extra-adrenal disease, but was also present in patients with metastases without SDHB mutations or those with metastases secondary to adrenal tumours. High risk of malignancy associated with SDHB mutations reflected large size and extra-adrenal locations of tumours, both independent predictors of metastatic disease. A plasma methoxytyramine above 0.2nmol/L or a tumour diameter above 5cm indicated increased likelihood of metastatic spread, particularly when associated with an extra-adrenal location. CONCLUSION: Plasma methoxytyramine is a novel biomarker for metastatic PPGLs that together with SDHB mutation status, tumour size and location provide useful information to assess the likelihood of malignancy and manage affected patients. 01 juli 2012

Commercialization of the media refers to the increasing importance of market considerations in the daily work of media professionals. Media organizations need to find ways to maintain the quality promise of their brands as an economically successful business. The increasing market orientation of media companies challenges their dedication to good journalism. The media brand identity approach is a good starting point from which to analyze this decision making in a commercialized and complex environment. By introducing the MBAC (“media, brands, actors, and communication”) model, a new approach is offered toward better theoretical understanding of brand identity-driven decision making. Responsibilities and tensions between the journalistic and the business side of news production are presented, and the complex environment in which decisions are made, implemented, and defended are elaborated on.

Mass-media coverage is important for both citizens and political actors: Citizens use the media as their main source of information for opinion formation. ‘Some minimal level of information facilitates the exercise of citizenship’ (Bennett and Iyengar 2008: 717). Political actors aim at influencing the judgement and voting decisions of citizens, but most of them have very limited resources (e.g. financial resources) to get into contact with their target audience by their own organizational channels (e.g. party newsletters, etc.) or by paid media (e.g. political advertising). Thus, getting access to the editorial content of mass media by means of political public relations messages (earned media) is a crucial strategy to convey campaign messages to the citizens. The success of this strategy heavily depends on the way the media treat political public relations messages, and on the degree of professionalization of PR in political organizations.

Media are a crucial actor in direct-democratic societies. In the political communication process, as we saw in Chapter 1 (see also Buchanan 2001), media basically transport political messages from political actors to citizens. Media thus serve democracy as an important and indispensable instrument. However, commercialization of the media, a shift from the dominance of journalistic values to the dominance of financial considerations, and a shift from citizen orientation to consumer orientation, puts in danger the democratic functions of the media.

Die Ansichten daruber, was das Unterhaltungsfernsehen zu Public Value beitragen kann, gehen weit auseinander. Von den Rezipienten wird Unterhaltung genutzt, um in Phantasiewelten einzutauchen, die die Grenzen des Alltags aufheben. Der Zuseher kann sich „Wunsche und Bedurfnisse befriedigen, die ihm die alltagliche Lebenswelt mit ihren spezifischen Relevanzbereichen versagt“ (Stumm/Dorsch-Jungsberger 1996: 132). Je nach Weltanschauung kann diese Entspannung nun als funktional und gesellschaftlich gewunscht oder als dysfunktional und gesellschaftsschadlich angesehen werden. Die Diskussion um den Wert von Unterhaltung ist so alt wie das offentliche Fernsehen. Lange bevor der Begriff Public Value als Beschreibung der eigenen Leistung in das Vokabular von Verantwortlichen bei Public-Service-Sendern aufgenommen wurde (vgl. Freedman 2008) wurde gestritten, was der Beitrag der Unterhaltung zum Auftrag der offentlichen Sender sein konnte. So haben sich z. B. die Mainzer Tage der Fernsehkritik 1970 der Unterhaltung im Fernsehen gewidmet. Der damalige Geschaftsfuhrer der Bavaria Studios propagierte eine paternalistische Meritorik (vgl. Erlei 1992) wenn er erklart, es sei die Aufgabe der Programmverantwortlichen, das Publikum „daran zu hindern, das zu wollen, was seinem Nutzen, seiner Freiheit und seiner Orientierung in die Zukunft entgegensteht“ (Gottschalk 1971: 109). Scheuch (1971) dagegen glaubt nicht, dass der offentliche Rundfunk durch ein erzieherisch gemeintes Unterhaltungsprogramm die Bedurfnisse des Publikums lenken konne.1 Entsprechend wundert er sich uber damals unter Medienschaffenden populare Forderungen, wonach der offentliche Rundfunk sich als Anstalt zu Beforderung von Umsturzen verstehen sollte, indem die Rezipienten der Berieselung entrissen werden sollten.

In this paper a new tool to assess viscoelastic and dielectric properties of human fluids is presented. Shear horizontal polarized surface acoustic waves (SH-SAW) are used to detect the viscoelastic properties of coagulating blood and blood plasma samples. One-port SAW resonators, with fundamental modes of 85, 170 und 340 MHz were developed. Additionally, their electrode structures can be used simultaneously to detect the dielectric behavior of the whole system by impedance spectroscopy while the frequency ranges from kHz to MHz. The combination of both methods offers the detection of clinical relevant blood parameters like the blood coagulation time and the hematocrit value within one measurement.

Im Beitrag wird nicht allein Online-Werbung analysiert, sondern auf die Besonderheiten der Online-Kommunikation im Hinblick auf Werbung eingegangen. Damit ist es moglich, auch werbliche Phanomene, die sich dem traditionellen Werbeprozess entziehen und das Potenzial der Online-Kommunikation innovativ einsetzen, zu thematisieren. Online- Medien konnen als Werbetrager aufgefasst werden, wenn sie lediglich zum Transport klassischer Werbemittel (Anzeige oder Spot) benutzt werden und insofern nur als weitere Distributionsplattform neben oder erganzend zu anderen fungieren. Die Besonderheiten der Online-Kommunikation beeinflussen aber bereits diese klassische Werbetragerfunktion, indem sie den ublichen Prozess und die ublichen Formen z.B. durch die aktive Einbeziehung der Werbenutzer verandern. Die Besonderheiten der Online- Kommunikation, Interaktivitat, Personalisierung, Netzwerke und Mobile Medien/ Endgerate sind auch fur die Werbung ausschlaggebend und werden deshalb jeweils hinsichtlich ihrer Auswirkungen auf Werbeform und Werbebotschaft sowie auf den werblichen Kommunikationsprozess diskutiert. Nach einer kurzen Ubersicht uber Werbeformate im Internet werden als Beispiele fur die Potenziale aber auch die Risiken, die sich aus der Kombination von Online-Kommunikation und Werbung ergeben, Word-of- Mouth bzw. virales Marketing und Communities behandelt.

In diesem Kapitel wird die wissenschaftliche Herangehensweise an das Phanomen Werbung skizziert. Im Alltag zeigt sich Werbung omniprasent, von facettenreicher Komplexitat und in vielschichtigen Erscheinungsformen und Vernetzungen. Entsprechend umfassend gestaltet sich die wissenschaftliche Auseinandersetzung mit dem Thema. Sichtet man die Literatur zur Werbung, sieht man sich mit einer nahezu unuberschaubaren Fulle und Bandbreite konfrontiert. Es nimmt daher kaum Wunder, dass Werbung nicht nur studentische Qualifikationsarbeiten inspiriert, sondern auch unterschiedlichste Wissenschaftsdisziplinen beschaftigt. Diese werden mit ihrem Bezug zur Werbung kurz vorgestellt, bevor der Stellenwert der Werbung als Forschungsgegenstand der Publizistik-und Kommunikationswissenschaft thematisiert wird.

Die vorhergehenden Kapitel konnten zeigen, dass es vielfaltige wissenschaftliche Erkenntnisse zur Werbung gibt, obwohl Werbung in der Publizistikund Kommunikationswissenschaft weder ein in sich geschlossenes Forschungsfeld darstellt noch als Kernbereich verankert ist. Gleichwohl ist Werbung von den gesellschaftspolitischen, okonomischen, technologischen, soziokulturellen und medialen Entwicklungen der Medien- und Informationsgesellschaft betroffen und wirkt an diesen mit. Dies fuhrt sowohl zur Aus-als auch zur Entdifferenzierung der Werbung, so dass die begriffliche Systematik nur eine vorlaufige sein kann. Auch der Einblick in die Geschichte belegt, dass Werbung ein wichtiger Indikator fur den gesellschaftlichen Wandel ist, wenn die entsprechenden Selektionslogiken berucksichtigt werden. Die aktuelle Relevanz der Werbung lasst sich einerseits aus den entsprechenden Kennzahlen interpretieren und grundet andererseits auf ihrer kontinuierlichen Systematisierung, Professionalisierung und Autonomisierung.

Werbung kann nicht fur sich allein betrachtet werden. Sie ist in ihrer Relevanz und Bedeutung sowie in Ablauf und Ergebnissen wesentlich von allgemeinen Rahmenbedingungen abhangig und beeinflusst diese mit. Die Rahmenbedingungen sind ihrerseits einer mehr oder weniger dynamischen Entwicklung unterworfen und von einzelnen herausragenden Ereignissen gepragt. Auf diesen Meta-Prozessen aufbauend ist Werbung in einen historischen Kontext eingebettet, und ihr jeweiliger Status, ihre Prozesse, Akteure und Ergebnisse sind von diesem mitbestimmt.