Evaluation of coagulation factors and platelet function from an off-line modified ultrafiltration technique for post-cardiopulmonary bypass circuit blood recovery.

Modified ultrafiltration (MUF) is a technique that hemoconcentrates residual CPB circuit blood and the patient at the same time. Hemoconcentration and MUF are Class 1-A recommendations in the anesthesia and surgical blood conservation guidelines. This study evaluated the off-line MUF process of the Hemobag (HB, Global Blood Resources, Somers, CT, USA) to quantitate coagulation factor levels, platelet (PLT) count and function in one facility and cellular growth factor concentrations of the final product that were transfused to the patient in another facility In two cardiac surgery facilities, after decannulation, the extracorporeal circuit (ECC) blood from 22 patients undergoing cardiac surgery was processed with the HB device. In eleven patients from the first facility by the study design, blood samples for coagulation factor levels and PLT aggregation were drawn from the reservoir of the MUF device pre- and post-processing. The samples (n = 11) were sent to a reference laboratory where testing for prothrombin time (PT), international normalized ratio (INR), activated partial thromboplastin time (aPTT), reptilase time, fibrinogen, clotting factors II, V, VII, VIII, IX, X, ADAMTS-13, protein C, protein S, antithrombin III, von Willebrand Factor (vWF), and platelet (PLT) aggregation were performed. A portion of the final concentrated HB blood samples (n = 5-10) from the second facility by design were evaluated for transforming and platelet-derived cellular growth factor concentrations. On average, approximately 800 - 2000 mls of whole blood were removed from the ECC post-CPB for processing in the HB device. After processing, there was, on the average, approximately 300 - 950 mls of concentrated whole blood salvaged for reinfusion. The PT and INR were significantly lower in the post-processing product compared to the pre-processing samples while the aPTT times were not significantly different. All coagulation factors and natural anti-coagulants were significantly increased in the final product. The PLT number, although increased by 24%, was not statistically significant. While PLT function assays showed a statistically significant decrease in the levels post-processing, there was substantial platelet function in the MUF product. Overall, the decrease in function was in the range of 10% to 15%. Final product PDGF-αß and TGF-ß1 averaged 11,048 and 2,040 pg/ml, respectively. In these two case series, (ECC) circuit blood concentrated using the HB device showed coagulation studies with significantly lower PT and INR and significantly increased levels of all clotting factors. The findings are similar to trends reported in other studies utilizing conventional MUF and the HB. Functioning platelets remain in the final product, with growth factor concentrations similar to some methods employed to create platelet concentrates to enhance coagulation. Based on the ability of the HB off-line MUF procedure to concentrate circuit blood, the clinical utility of the HB device to decrease allogeneic blood product exposure should be evaluated in a prospective randomized clinical trial.

Paving the way for transgenic schistosomes.

In parasitological research, significant progress has been made with respect to genomics and transcriptomics but transgenic systems for functional gene analyses are mainly restricted to the protozoan field. Gene insertion and knockout strategies can be applied to parasitic protozoa as well as gene silencing by RNA interference (RNAi). By contrast, research on parasitic helminthes still lags behind. Along with the major advances in genome and transcriptome analyses e.g. for schistosomes, methods for the functional characterization of genes of interest are still in their initial phase and have to be elaborated now, at the beginning of the post-genomic era. In this review we will summarize attempts made in the last decade regarding the establishment of protocols to transiently and stably transform or transfect schistosomes. Besides approaches using particle bombardment, electroporation or virus-based infection strategies to introduce DNA constructs into adult and larval schistosome stages to express reporter genes, first approaches have also been made in establishing protocols based on soaking, lipofection, and/or electroporation for RNA interference to silence gene activity. Although in these cases remarkable progress can be seen, the schistosome community eagerly awaits major breakthroughs especially with respect to stable transformation, but also for silencing or knock-down strategies for every schistosome gene of interest.

The undue influence of shape and weight on self-evaluation in anorexia nervosa, bulimia nervosa and restrained eaters: a combined ERP and behavioral study.

BACKGROUND: Current theories and nosology assume that the self-evaluation (SE) of individuals with eating disorders (EDs) is unduly influenced by body shape and weight. However, experimental data supporting this link are scarce, and it is not specified which subdomains of SE might be affected. METHOD: We studied patients with anorexia nervosa (AN), bulimia nervosa (BN) and healthy controls (HC) with an affective priming (AP) procedure (Study 1) to unveil explicit and implicit associations between shape/weight and SE. We used weight/shape-related prime sentences, complemented by affectively congruent and incongruent target words from two SE domains. AP effects were assessed by event-related potentials (ERPs), reaction times (RTs) and subjective ratings. The ratings were also assessed (Study 2) in undergraduate restrained (RES) and unrestrained eaters (UNRES). RESULTS: Study 1 demonstrated stronger AP effects in both ED groups compared to HC on RTs and subjective ratings. ERPs showed AP effects only in the BN group. Restrained eaters showed similar, albeit less pronounced, priming effects on subjective ratings. CONCLUSIONS: ED patients associate shape/weight concerns with the non-appearance-related SE domains of interpersonal relationships and achievement/performance. These associations seem to be encoded deeper in BN patients relative to the other groups. Links between shape/weight and SE explain how body dissatisfaction impacts on self-esteem and mood in ED. The existence of similar associations in restrained eaters supports a continuum model according to which increasing associations between shape/weight and SE go along with increasing levels of ED symptoms.

Schistosoma mansoni: signal transduction processes during the development of the reproductive organs.

Among the topics of considerable interest concerning our understanding of the unusual biology of schistosomes is the sexual maturation of the female. The identification of genes coding for signal transduction proteins controlling essential steps of the pairing-dependent differentiation of the reproductive organs, vitellarium and ovary will help to substantiate our knowledge about this unique parasite. Furthermore, such signalling proteins could be potential targets to interfere with the development of this parasite to combat schistosomiasis since its pathology is caused by the eggs. This review summarises first post-genomic steps to elucidate the function of gonad-specific signalling molecules which were identified by homology-based cloning strategies, by in silico identification or by yeast two-hybrid interaction analyses, using a combination of novel techniques. These include the in vitro culture of adult schistosomes, their treatment with chemical inhibitors to block enzyme activity, the use of RNAi to silence gene function post-transcriptionally, and confocal laser scanning microscopy to study the morphological consequences of these experimental approaches. Finally, we propose a first model of protein networks that are active in the ovary regulating mitogenic activity and differentiation. Some of these molecules are also active in the testes of males, probably fulfilling similar roles as in the ovary.

Imatinib has a fatal impact on morphology, pairing stability and survival of adult Schistosoma mansoni in vitro.

Schistosomes cause bilharzia (schistosomiasis), one of the most prevalent parasitic diseases for human and animals worldwide. Praziquantel (PZQ) is the only widely used drug for treatment and control of this parasitemia. Since a vaccine is not yet available, and in light of emerging resistance against PZQ, the search for alternatives has high priority. Here we present that Imatinib, a compound used in human cancer therapy (Gleevec; STI-571), significantly affected schistosome morphology and physiology in vitro. Besides its negative effect on gonad development and pairing stability, Imatinib led to pathological alterations of the gastrodermis, which finally caused the death of the parasite.

Molecular characterisation of kappa-5, a major antigenic glycoprotein from Schistosoma mansoni eggs.

The major immunopathological consequences of infection with Schistosoma mansoni, a T helper type 2 response and granuloma formation leading to fibrotic tissue damage, are caused by the egg stage of the parasite. Three antigens of S. mansoni eggs, termed IPSE/alpha-1, omega-1 and kappa-5, have been found to be the primary targets of the egg-directed antibody response of the host. Here, we report on the isolation, cloning and characterisation of kappa-5. Apart from an uncharacterised mRNA sequence in S. japonicum, no significant similarities of kappa-5 to known sequences from other species were found. In contrast to IPSE/alpha-1 and omega-1, which have been found only in eggs, kappa-5 was present in miracidia as well as in eggs at the mRNA and protein levels. In eggs, isoforms of kappa-5 were observed with both three and four fully occupied N-glycosylation sites, while in miracidia only one isoform with four N-glycans could be detected. Interestingly, in Western blots sera from S. mansoni-infected Africans were reactive against kappa-5 with IgE and IgG isotype antibodies, but against IPSE/alpha-1 and omega-1 only with IgG antibodies. The further characterisation of kappa-5 as one of the three major egg antigens should help to better understand the immunology and immunopathology of schistosomiasis.

Posterior and anterior components of force during bite loading.

Late anterior crowding of teeth has been associated with the anterior component of force (ACF) developed during biting. Possible physiologic mechanisms countering ACF, including the presence of a posterior component of force (PCF), are hypothesized. In this self-controlled study, 60 subjects aged 27.05+/-3.9 years were examined for ACF and PCF that were calculated as the change in tightness of a mandibular dental contact points from non-biting to biting state. Both ACF and PCF were found to develop simultaneously. However, the PCF was 4-7 folds smaller than the ACF (p<0.001). The ACF progressively declined by 10-20 folds (p<0.001) from the posterior to anterior dentition. The lateral incisor-canine contact point had the greatest ACF decline (63-74%). ACF effect on the anterior dentition is counteracted by a protective mechanism consisted of PCF, progressive dissipation of ACF, and canine blockage.

Ribozyme targeting of HER-2 inhibits pancreatic cancer cell growth in vivo.

We have analysed HER-2 expression and function in pancreatic cancer cells to determine whether HER-2 has a rate-limiting role for pancreatic cancer cell growth in vitro and in vivo. To specifically assess HER-2 function, we used HER-2-targeted ribozymes expressed under the control of the tet-off promoter system. Six out of 11 human pancreatic cancer cell lines expressed all four epidermal growth factor (EGF)-receptor family members (HER-1 (EGF-R), HER-2, HER-3, and HER-4), including Panc89 cells. Expression of the ribozymes quenched endogenous HER-2 mRNA levels in Panc89 cells by approximately 40-60% which was reflected by a 40-50% reduction of the HER-2 surface glycoprotein. HER-2 depletion inhibited the in vitro proliferation rate by approximately 40% and decreased in vivo tumour growth by approximately 60% (P<0.05). Our study demonstrates for the first time a rate-limiting role for HER-2 in pancreatic cancer cell proliferation and suggests HER-2 targeting as a potential approach in pancreatic cancer therapy.

Use of intravascular ultrasound to compare effects of different strategies of lipid-lowering therapy on plaque volume and composition in patients with coronary artery disease.

BACKGROUND: We studied whether lipid-lowering therapy with atorvastatin (target LDL cholesterol [LDL-C] <100 mg/dL) compared with a moderate treatment regimen that used other lipid-lowering drugs led to a lesser progression of atherosclerosis and to different changes in plaque echogenicity in patients with coronary artery disease. METHODS AND RESULTS: This study was a 12-month, open-label, randomized, multicenter trial, which used serial 3D intracoronary ultrasound to calculate plaque volume and plaque echogenicity. After transcatheter therapy, 131 patients were randomized (atorvastatin n=65, usual care n=66). The target plaque had to be a minor lesion (ie, a diameter stenosis of <50% on angiography). After 12 months, mean LDL-C was reduced from 155 to 86 mg/dL in the atorvastatin group and from 166 to 140 mg/dL in the usual care group. Mean absolute plaque volume showed a larger increase in the usual care group compared with the atorvastatin group (usual care 9.6+/-28.1 mm(3), atorvastatin 1.2+/-30.4 mm(3); P=0.191). The hyperechogenicity index of the plaque increased to a larger extent for the atorvastatin group than for the usual care group, with a significant treatment effect for the percent change (atorvastatin 42.2%, usual care 10.1%; P=0.021). CONCLUSIONS: One year of lipid-lowering therapy to <100 mg/dL LDL-C most likely led to a slowdown of plaque growth of minor lesions. The significantly larger increase in plaque hyperechogenicity is most likely due to a change in plaque composition.

Comparative analysis of the EGF-receptor family in pancreatic cancer: expression of HER-4 correlates with a favourable tumor stage.

BACKGROUND: Of major interest for a better molecular understanding of pancreatic cancer is the EGF receptor family. While HER-1 (EGF-receptor) and HER-2 have been extensively studied, little is known about the clinical significance of HER-3 and especially HER-4 expression. METHODS: We investigated the expression of HER-1, HER-2, HER-3 and HER-4 in 11 pancreatic cancer cell lines using FACS-analysis and determined expression and overexpression of these receptors in 24 pancreatic cancer specimens. Therefore, we used two different immunostaining techniques: a highly sensitive streptavidin-biotin method showed receptor expression while an approximatly 10-fold less sensitive indirect immunperoxidase technique determined receptor-overexpression. RESULTS: HER-1 and HER-2 were expressed by all 11 pancreatic cancer cell lines, HER-3 was found in 82% and HER-4 in 54% of the cell lines. Low levels of HER-1, HER-2 and HER-3 were detected in all tumor samples but overexpression was only found in 33%, 25% and 50% of the cases, respectively. HER-4 was expressed by 37% of the tumor specimens but overexpression was seen in one patient only. HER-1 and HER-2 overexpression increased in parallel with the tumor stage and R0-resected tumors showed significantly less often overexpression compared to R1/R2 resected tumors (p < 0.05). In contrast, 54% of the R0-resected vs. 18% of the R1/R2 resected tumors showed HER-4 expression (p = 0.07) and HER-4 was exclusively found in non-metastatic tumors (p = 0.0149). CONCLUSION: Our data indicate that HER-1 and HER-2 overexpression contributes to a more aggressive phenotype. In contrast, the lack of HER-4 expression might increase the metastatic capacity of pancreatic cancer cells.

Combined thallium-201 and dynamic iodine-123 iodophenylpentadecanoic acid single-photon emission computed tomography in patients after acute myocardial infarction with effective reperfusion.

BACKGROUND: Considerable derangements of energy metabolism are to be expected during ischemia and reperfusion. In ischemic myocardium, the oxidative degradation of carbohydrates is shifted toward the anaerobic production of lactate and the oxidation of fatty acids is suppressed. HYPOTHESIS: The aim of this study was to examine the uptake and metabolism of iodine-123 (123I) iodophenylpentadecanoic acid (IPPA) in stunned myocardium. METHODS: In 15 patients, SPECT with 201Tl and 123I IPPA as well as echocardiography with low-dose dobutamine stimulation were performed 12 +/- 5 days after myocardial infarction with reperfusion. Follow-up echocardiography was carried out 24 +/- 8 days later for documentation of functional improvement. Uptake of 201Tl and 123I IPPA were obtained in five left ventricular segments, and dynamic SPECT imaging was used for calculation of the fast and the slow components of the biexponential myocardial 123I IPPA clearance. RESULTS: Wall motion improved in 14 of 26 dysfunctional segments (54%). Stunned segments were characterized by a reduced 123I IPPA extraction, a shorter half-life of the fast, and a longer half-life of the slow clearance component. All parameters of the combined 201Tl/123I IPPA study predicted functional recovery with similar accuracies (area under the receiver operator characteristic curves between 0.68 and 0.76; p = NS). Analysis of 201Tl uptake alone could not predict functional recovery in this study. CONCLUSIONS: Stunned myocardium is characterized by a disturbance of fatty acid metabolism. For prediction of functional improvement, 123I IPPA imaging added significant diagnostic information.

Bcl-XL protects pancreatic adenocarcinoma cells against CD95- and TRAIL-receptor-mediated apoptosis.

In this study we sought to clarify the role of the proapoptotic potential of mitochondria in the death pathway emanating from the TRAIL (APO-2L) and CD95 receptors in pancreatic carcinoma cells. We focused on the role of the Bcl-2 family member Bcl-XL, using three pancreatic carcinoma cell lines as a model system, two of which have high (Panc-1, PancTuI) and one has low (Colo357) Bcl-XL expression. In these cell lines, the expression of Bcl-XL correlated with sensitivity to apoptosis induced by TRAIL or anti-CD95. Flow cytometric analysis revealed cell surface expression of TRAIL-R1 and TRAIL-R2 on PancTuI and Colo357, and TRAIL-R2 on Panc-1 cells. In Colo357 cells retrovirally transduced with Bcl-XL, caspase-8 activation in response to treatment with TRAIL or anti-CD95 antibody was not different from parental cells and EGFP-transfected controls, however, apoptosis was completely suppressed as measured by the mitochondrial transmembrane potential deltapsim, caspase-3 activity (PARP cleavage) and DNA-fragmentation. Inhibition of Bcl-XL function by overexpression of Bax or administration of antisense oligonucleotides against Bcl-XL mRNA resulted in sensitization of Panc-1 cells to TRAIL and PancTuI cells to anti-CD95 antibody-induced cell death. The results show that Bcl-XL can protect pancreatic cancer cells from CD95- and TRAIL-mediated apoptosis. Thus, in these epithelial tumour cells the mitochondrially mediated 'type II' pathway of apoptosis induction is not only operative regarding the CD95 system but also regarding the TRAIL system.

[Experiences with ambulatory cardiologic phase II rehabilitation]. / Erfahrungen mit der ambulanten kardialen Rehabilitation der Phase II.

The phase II cardiac rehabilitation in Germany differs markedly from other European countries and the USA. Most of the patients enter a 3-week full residential program. In contrast we developed an outpatient phase II cardiac rehabilitation program. Since 1979 we treated more than 8,500 patients with different indications (i.e. after myocardial infarction, coronary bypass surgery, valve replacement and reconstruction). Patients with a daily commuting time over 60 minutes are not suitable for outpatient rehabilitation. Our model corresponds to the German intrahospital rehabilitation. The rehabilitation is carried out in 3 weeks offering approximately 66 hours of therapy. Groups of 8 patients with a similar level of physical capacity stay together during the rehabilitation. A comprehensive program with exercise training, physical therapy, psychological support, education in life style changes and risk factor modification has been developed. The compliance of the patients as well as the acceptance by the family are excellent. Long-lasting reduction in LDL cholesterol levels and increments in work-load capacities have been demonstrated. A high percentage of patients returned to work. Cost analysis demonstrates a reduction up to 40% in comparison to the full residential program. Therefore the outpatient phase II cardiac rehabilitation is a good alternative especially in urban areas.

Psychosocial characterization of patients with atopic dermatitis in conventional versus alternative-medical therapy.

OBJECTIVE: Recording of psychosocial differences between patients with atopic dermatitis (AD) undergoing conventional versus alternative-medical treatment as indication of a center-related selection effect. STUDY DESIGN: In this prospective comparative study, inpatients of an alternative-medical clinic and of the University Dermatology Clinic Freiburg were given standardized questionnaires on psychosocial characteristics in the areas of disease-specific stress, social support, and coping with the disease. PATIENTS: Fifty-nine consecutive AD patients were recruited at the alternative-medical clinic and 79 AD patients at the University Dermatology Clinic. Fifty-five (93.2%) and 73 (92.4%) patients respectively returned completed questionnaires. RESULTS: The two treatment groups differed markedly from one another with respect to psychosocial parameters. Patients of the alternative-medical clinic showed greater disease-specific stress, lower social integration, and higher values in the coping scales 'depressive coping' and 'religious conviction and search for meaning'. Compared to patients with acne vulgaris, both patient groups were under significantly greater disease-specific stress. The factor 'social integration' was determined by logistical regression analysis to be the most important differentiation characteristic between the groups. CONCLUSIONS: Patients under alternative-medical treatment may show significant psychosocial differences in comparison with conventionally treated patients. In comparative studies between conventional and alternative-medical therapy centers, selection effects must be assumed and taken into account when evaluating the study.

Alveolar and skeletal dimensions associated with overbite.

The aim of this study was to investigate whether in the maxilla and in the mandible the structure of the anterior medial sagittal alveolar and basal bone is related to the overbite. A total of 460 untreated adult subjects were divided into four groups with either deep bite, normal overbite, end-to-end bite, or open bite and were compared. The overbite, lower face height, and anterior alveolar and basal midsagittal cross-sectional areas from the maxilla and the mandible were assessed on lateral cephalometric radiographs. An index was calculated, dividing the sagittal by the vertical dimension of the midsagittal cross-sectional area. A deeper bite coincided with smaller lower face height, larger alveolar and basal areas, and a more widened shape of the symphysis. If the lower face height was introduced as a covariable, the open bite group showed significantly smaller maxillary and mandibular alveolar and basal cross-sectional areas compared with the end-to-end group, the normal overbite group, or the deep bite group. Vertical variation of the overbite probably coincides with a relative hyperdevelopment or hypodevelopment of the symphysis.

Alveolar and skeletal dimensions associated with lower face height.

In this study, the relationships between the lower face height and the structure of the frontal alveolar and basal bone were investigated. The areas and the dimensions of the anterior alveolar and basal midsagittal cross-sectional bone from the maxilla and the mandible were recorded on lateral cephalograms from 460 untreated adults. An index was calculated dividing the sagittal by the vertical dimension of the midsagittal cross-sectional area. The subjects with a normal overbite between 0.5 and 4 mm (N=165) were divided into three groups according to the lower face height. A larger lower face height coincided with a larger maxillary alveolar and basal area and with a smaller mandibular alveolar index. Correlations between the lower face height and the maxillary alveolar index and the mandibular alveolar and basal area were low. It is concluded that long-faced subjects have a large mandibular alveolar height, which is more associated with a narrowed shape than with a large volume of the symphysis.

Does dobutamine stress echocardiography induce damage during viability diagnosis of patients with chronic regional dysfunction after myocardial infarction?

Experimental hibernating-model investigations of animals have shown that myocardial necrosis can be induced by longer-term intracoronary dobutamine infusion. This study was designed to determine whether myocardial infarction could be ascertained in patients with chronic regional wall motion abnormalities and greater than 75% stenosis in the supplying coronary artery through dobutamine stress echocardiography. Twenty patients with coronary artery disease and regional resting wall motion abnormalities were examined with a standard dobutamine protocol (5 to 50 microg/kg/min). Exclusion criteria were an acute coronary syndrome, severe heart failure, and severe hypertension. Creatine kinase (CK, CKMB), myoglobin, and troponine-I were measured before and at each of the first 7 hours after beginning of infusion. Fourteen of these 20 patients exhibited viable myocardium. The serum markers CK, CKMB, myoglobin, and troponin-I demonstrated no increase beyond the reference range, suggesting that with this protocol, no myocardial necrosis was induced.

Safety of intracoronary ultrasound imaging in patients with acute myocardial infarction.

In 103 patients with acute myocardial infarction, intracoronary ultrasound imaging (ICUS) was performed before and after percutaneous transluminal coronary angioplasty (PTCA) with a pre-PTCA success rate of 79 of 103 patients (76.7%), post-PTCA rate of 88 of 103 patients (85.4%), and a reversible subacute occlusion rate after initial ICUS of 3.9%. Time consumption was 7 +/- 1 minute for pre-PTCA ICUS and 3 +/- 1 minute for post-PTCA ICUS.

Stress echocardiography in special groups: in women, in left bundle branch block, in hypertension and after heart transplantation.

The non-invasive diagnosis of coronary artery disease by exercise electrocardiography is less accurate in women than in men, with a high rate of false-positive results in women. In contrast, recent studies have demonstrated that stress echocardiography in women is more accurate than exercise echocardiography and that the significantly higher specificity of stress electrocardiography may have the benefit of avoiding unnecessary angiography in women. Exercise-induced changes in the electrocardiogram are non-diagnostic in the presence of left bundle branch block or basal ST changes. In these patients, stress echocardiography can be used instead of conventional scintigraphy for the detection of coronary artery disease, but further echocardiographic studies are needed to confirm the promising results. Exercise electrocardiography and exercise echocardiography have been reported to be disappointing in the early detection of cardiac allograft vasculopathy after heart transplantation, and dobutamine stress echocardiography overestimates the incidence of angiographic evidence of cardiac allograft vasculopathy. However, compared to intravascular ultrasound imaging, dobutamine stress echocardiography seems to be a suitable non-invasive method for detecting cardiac allograft vasculopathy.