[Fostering organ donation in the intensive care unit : A best practice example]. / Förderung der Organspende auf der Intensivstation : Ein Best-Practice-Beispiel.

We report on the development and implementation of a bundle of measures with which we increased the number of post-mortem organ donations performed in our hospital from 2 per year to 10 per year within 5 years.

Determination of mechanical and rheological properties of a cell-loaded peptide gel during ECM production.

The development of an injectable biomaterial that supports cell survival and maintains or promotes nucleus pulposus (NP) phenotype could aid delivery of cells to degenerated NPs causing low back pain. Mesenchymal cells were loaded and grown in a synthetic peptide gel, PuraMatrix®. Cells were observed within the gels over 0-28 days, and accumulation of glycosaminoglycans were detected by histological staining. The mechanical properties of the cell-loaded constructs, and the change of the mechanical properties were studied using stress relaxation of the gels under compression and confinement. The PuraMatrix® gel was shown to relax fast on compression indicating that the fluid could easily flow out of the gel, and thus indicating the presence of large pores/voids. The presence of these pores/voids was further supported by high mobility of dextran molecules, determined using fluorescence recovery after photo bleaching. The stress required to deform the cell-loaded constructs to a specific strain increases at day 21, at which point the presence of glycosaminoglycans within the cell-loaded constructs was also observed. The results provide evidence of changes in mechanical properties of the PuraMatrix® matrix upon excretion of the extracellular matrix by the cells.

Single particle raster image analysis of diffusion for particle mixtures.

Recently we complemented the raster image correlation spectroscopy (RICS) method of analysing raster images via estimation of the image correlation function with the method single particle raster image analysis (SPRIA). In SPRIA, individual particles are identified and the diffusion coefficient of each particle is estimated by a maximum likelihood method. In this paper, we extend the SPRIA method to analyse mixtures of particles with a finite set of diffusion coefficients in a homogeneous medium. In examples with simulated and experimental data with two and three different diffusion coefficients, we show that SPRIA gives accurate estimates of the diffusion coefficients and their proportions. A simple technique for finding the number of different diffusion coefficients is also suggested. Further, we study the use of RICS for mixtures with two different diffusion coefficents and investigate, by plotting level curves of the correlation function, how large the quotient between diffusion coefficients needs to be in order to allow discrimination between models with one and two diffusion coefficients. We also describe a minor correction (compared to published papers) of the RICS autocorrelation function.

Single particle raster image analysis of diffusion.

As a complement to the standard RICS method of analysing Raster Image Correlation Spectroscopy images with estimation of the image correlation function, we introduce the method SPRIA, Single Particle Raster Image Analysis. Here, we start by identifying individual particles and estimate the diffusion coefficient for each particle by a maximum likelihood method. Averaging over the particles gives a diffusion coefficient estimate for the whole image. In examples both with simulated and experimental data, we show that the new method gives accurate estimates. It also gives directly standard error estimates. The method should be possible to extend to study heterogeneous materials and systems of particles with varying diffusion coefficient, as demonstrated in a simple simulation example. A requirement for applying the SPRIA method is that the particle concentration is low enough so that we can identify the individual particles. We also describe a bootstrap method for estimating the standard error of standard RICS.

Sex peptide of Drosophila melanogaster males is a global regulator of reproductive processes in females.

Seminal fluid proteins (Sfps) alter female behaviour and physiology and can mediate sexual conflict. In Drosophila melanogaster, a single Sfp, the sex peptide (SP), triggers remarkable post-mating responses in females, including altered fecundity, feeding, immunity and sexual receptivity. These effects can favour the evolutionary interests of males while generating costs in females. We tested the hypothesis that SP is an upstream master-regulator able to induce diverse phenotypes through efficient induction of widespread transcriptional changes in females. We profiled mRNA responses to SP in adult female abdomen (Abd) and head+thorax (HT) tissues using microarrays at 3 and 6 h following mating. SP elicited a rich, subtle signature of temporally and spatially controlled mRNAs. There were significant alterations to genes linked to egg development, early embryogenesis, immunity, nutrient sensing, behaviour and, unexpectedly, phototransduction. There was substantially more variation in the direction of differential expression across time points in the HT versus Abd. The results support the idea that SP is an important regulator of gene expression in females. The expression of many genes in one sex can therefore be under the influence of a regulator expressed in the other. This could influence the extent of sexual conflict both within and between loci.

Computerized occlusion rating: a superior predictor of aneurysm rebleeding for ruptured embolized aneurysms.

BACKGROUND AND PURPOSE: The initial angiographic occlusion rate is the strongest predictor of later rebleeding in previously ruptured coil-embolized cerebral aneurysms. Angiographic estimations of aneurysmal occlusion rates are, however, subjective in nature and confounded by methodologic problems. COR has been developed, and its superiority has been experimentally established to overcome subjective bias. The purpose of this study was to assess the clinical value of COR as a more objective predictor of aneurysm rebleeding when compared with SOR as described in the Raymond Classification. MATERIALS AND METHODS: We applied COR in a consecutive series of 249 patients. Two DSA projections were selected independently by 2 blinded investigators. In cases of disagreement on the selected projections, a consensus decision was obtained. SOR were determined by 2 independent observers according to the Raymond classification. COR was measured by 2 blinded investigators. Interobserver variations were determined for SOR and COR. COR results were compared with SOR results and stratified as 100%, 99.9%-90%, 89.9%-70%, and <70% occlusion. SOR and COR were evaluated as predictors for aneurysm rebleeding. RESULTS: Seven aneurysms rebled (2.8%; follow-up, 59 ± 35 months). In 20.9% of all cases, DSA selection was performed by consensus evaluations. Interobserver variations were statistically significant for SOR (P = .0030) but not for COR (P = .3517). Compared with COR, SOR overestimated the degree of aneurysmal occlusion in 81.9% of all cases. Only COR predicted rebleeding (P = .0162). CONCLUSIONS: Unacceptable interobserver variations were shown for the standard SOR estimations. COR substantially reduced the impact of subjective bias. COR may, therefore, serve as an easily applicable more objective predictor of aneurysm rerupture. The remaining bias of COR, caused by 2D image analysis, may be overcome by use of direct 3D measurements.

Prognostic significance of L1CAM in ovarian cancer and its role in constitutive NF-κB activation.

BACKGROUND: Overexpression of L1-cell adhesion molecule (L1CAM) has been observed for various carcinomas and correlates with poor prognosis and late-stage disease. In vitro, L1CAM enhances proliferation, cell migration, adhesion and chemoresistance. We tested L1CAM and interleukin-1 beta (IL-1ß) expression in tumor samples and ascitic fluid from ovarian carcinoma patients to examine its role as a prognostic marker. PATIENTS AND METHODS: We investigated tumor samples and ascitic fluid from 232 serous ovarian carcinoma patients for L1CAM by enzyme-linked immunosorbent assay. L1CAM expression was correlated with pathoclinical parameters and patients' outcome. IL-1ß levels were measured in tumor cell lysates. Ovarian cancer cell lines were analyzed for the contribution of L1CAM to IL-1ß production and nuclear factor 'kappa-light-chain-enhancer' of activated B-cells (NF-κB) activation. RESULTS: We observed that L1CAM-expressing tumors show a highly invasive phenotype associated with restricted tumor resectability at primary debulking surgery and increased lymphogenic spread. Soluble L1CAM proved to be a marker for poor progression-free survival and chemoresistance. In ovarian carcinoma cell lines, the specific knock-down of L1CAM reduces IL-1ß expression and NF-κB activity. CONCLUSIONS: L1CAM expression contributes to the invasive and metastatic phenotype of serous ovarian carcinoma. L1CAM expression and shedding in the tumor microenvironment could contribute to enhanced invasion and tumor progression through increased IL-1ß production and NF-κB activation.

Δ133p53 is an independent prognostic marker in p53 mutant advanced serous ovarian cancer.

BACKGROUND: We aimed to evaluate the clinical relevance of p53 and p73 isoforms that modulate the function of p53. METHODS: This prospective multicentre study included 154 patients with stage III and IV serous ovarian cancer. A functional yeast-based assay and subsequent sequencing were performed to analyse the p53 mutational status. Expression of p53 and p73 isoforms was determined using RT-qPCR. RESULTS: Δ133p53 expression constituted an independent prognostic marker for recurrence-free (hazard ratio=0.571, P=0.016, 95% CI: 0.362-0.899) and overall survival (hazard ratio=0.365, P=0.004, 95% CI: 0.182-0.731) in patients with p53 mutant ovarian cancer (n=121). High Δ40p53 expression was associated with favourable tumour grading (P=0.037) and improved recurrence-free survival (33.4 vs 19.6 months, P=0.029), but not overall survival (43.1 vs 33.6 months, P=0.139), in patients with p53 wild-type cancer (n=33). Neither the p53 mutational status nor p73 isoform expression possessed prognostic significance in the examined ovarian cancer cases. CONCLUSION: Δ133p53 expression was associated with prognosis in the vast majority of ovarian cancer cases, that is, patients with p53 mutant advanced serous carcinomas. Thus, our findings underline the importance of considering the complex p53 regulatory network.

Examinations on the prevalence of footpad lesions and breast skin lesions in B.U.T. Big 6 fattening turkeys in Germany. Part II: Prevalence of breast skin lesions (breast buttons and breast blisters).

This project involves the prevalence of breast skin alterations (breast buttons, blisters, purulent bursitis) in male and female fattened turkeys (British United Turkeys Big 6) from conventional intensive farming. In this case, 24 turkey farms across Germany and 11,860 turkeys (5,740 toms, 6,120 hens) were clinically examined, and 16,200 birds (8,400 hens, 7,800 toms) were evaluated on slaughter lines at 7 turkey abattoirs. Each turkey farm included examination of at least 3 consecutive fattening cycles. Especially in the meat examinations, breast skin alterations were a frequent discovery, particularly in the form of breast buttons. The turkey toms exhibited a prevalence of 27.15%, and the toms were affected significantly more frequently (P < 0.001) by these variations than the turkey hens (7.77%). It is possible that the higher BW of the male specimens leads to increased lying times and is therefore connected with more severe strain on the breast region. The increased fattening period of the turkey toms (slaughtering at wk 21 to 22 of life) vs. the female specimens (slaughtering at wk 15 to 17 of life) must be discussed as a cause. Breast blisters and bursitis sternalis were discovered with a prevalence of 7.36 and 1.24% in toms and more seldom in hens, at 0.30% and 0.15%. Large differences in prevalence were partially recognizable between individual farms as well as between the 3 fattening cycles on a farm. Breast buttons and breast blisters are pathological skin variations for which the etiology is multifactorial. Clear correlations between the prevalence of breast skin alterations and the manner of rearing the birds were not able to be determined. An improvement in this situation, which is considered very problematic from the perspective of bird protection and economics, may be achieved only via cooperation to achieve optimized rearing methods, management, and breeding practices.

Examinations on the prevalence of footpad lesions and breast skin lesions in British United Turkeys Big 6 fattening turkeys in Germany. Part I: prevalence of footpad lesions.

The present study demonstrates the prevalence of footpad lesions in turkeys and their level of expression in both live birds and in carcasses. In clinical investigations of 11,860 turkeys (5,740 males, 6,120 females) of the strain British United Turkeys Big 6, individuals of all the observed flocks showed alterations to the plantar skin. In general, the degree and severity of skin alterations were higher in the 16th week of life than in the 6th or 11th week of life. Epithelial necrosis, however, was already apparent in about 45% of individuals in the 6th week of life. At this point, deep lesions on the footpad surface were still rare findings. In the 11th week of life, such skin changes were apparent in 14.7% of males and 25.7% of hens. In general, the feet of male turkeys were less affected by footpad damage than those of females in the subsequent fattening period. In hens, the prevalence of lesions of the footpads at wk 16 of age was 60.0%, almost twice as high as in males of the same age group (33.8%). A comparison of the findings from the right and left feet showed that the state of the 2 extremities was correlated (Spearman correlation: r = 0.830). During the visual inspection of 16,200 slaughtered turkeys (7,800 males, 8,400 hens), almost all carcasses studied showed alterations to the footpad skin. Only 2.1% of males and 0.6% of females were without lesions. The most frequently observed lesions measured up to 2 cm in diameter (males: 59.2%; hens: 57.7%). Pronounced skin lesions of the footpad (males: 21.1%; females: 29.5%) and necrosis of the superficial epithelium (males: 17.5%; females: 12.1%) were less common. Foot abscesses were diagnosed only sporadically (males: 0.1%; females: 0.1%).

Assessment of left ventricular volumes, ejection fraction and mass. Comparison of model-based analysis of ECG-gated (99m)Tc-SPECT and ¹8F-FDG-PET.

AIM: We compared and delineated possible differences of model-based analysis of ECG-gated SPECT using (99m)Tc-sestamibi (Tc-SPECT) with ECG-gated ¹8F-fluorodeoxyglucose-PET (FDG-PET) for determination of end-diastolic (EDV) and end-systolic (ESV) cardiac volumes, left ventricular ejection fraction (LVEF), and myocardial mass (LVMM). PATIENTS, METHODS: 24 patients (21 men; age: 54±12years) with coronary artery disease underwent Tc-SPECT and FDG-PET imaging for evaluation of myocardial perfusion and viability. By using model-based analysis EDV, ESV, LVEF and LVMM were calculated from short axis images of both Tc-SPECT and FDG-PET. RESULTS: Left ventricular volumes by Tc-SPECT and FDG-PET were 176±60 ml and 181±59 ml for EDV, and 97±44 ml and 103±45 ml for ESV respectively, LVEF was 47±8% by Tc-SPECT and 45±9% by FDG-PET. The LVMM was 214±40 g (Tc-SPECT) and 202±43 g (FDG-PET) (all p = NS, paired t-test). A significant correlation was observed between Tc-SPECT and FDG-PET imaging for calculation of EDV (r = 0.93), ESV (r = 0.93), LVEF (r = 0.83) and LVMM (r = 0.72). CONCLUSION: ECG-gated Tc-SPECT and FDG-PET using two tracers with different characteristics (perfusion versus metabolism) showed close agreement concerning measurements of left ventricular volumes, contractile function and myocardial mass by using a model-based analysis.

Alternative splicing of p53 and p73: the novel p53 splice variant p53delta is an independent prognostic marker in ovarian cancer.

Similar to p73, the tumor suppressor gene p53 is subject to alternative splicing. Besides p53DeltaE6 and p53beta, we identified p53zeta, p53delta and p53varepsilon, arising from alternative splicing of exon 6 and intron 9, respectively. p53 splice variants were present in 18 of 34 ovarian cancer cell lines (52.9%) and 134 of 245 primary ovarian cancers (54.7%). p53delta expression was associated with impaired response to primary platinum-based chemotherapy (P=0.032). Also, p53delta expression constituted an independent prognostic marker for recurrence-free and overall survival (hazard ratio 1.854, 95% confidence interval 1.121-3.065, P=0.016; and hazard ratio 1.937, 95% confidence interval 1.177-3.186, P=0.009, respectively). p53beta expression was associated with adverse clinicopathologic markers, that is, serous and poorly differentiated cancers (P=0.002 and P=0.008, respectively), and correlated with worse recurrence-free survival in patients exhibiting functionally active p53 (P=0.049). DeltaN'p73 constituted the main N-terminally truncated p73 isoform and was preferentially found in ovarian cancer cell lines showing functionally active p53, supporting our hypothesis that N-terminally truncated p73 isoforms can alleviate the selection pressure for p53 mutations by the inhibition of p53 protein function.

Rheo-NMR studies of the behavior of a nematic liquid crystal in a low-shear-rate regime: the transition from director alignment to reorientation.

Deuterium NMR spectroscopy has been used to study the director dynamics of the nematic liquid-crystal system cetyl trimethylammonium bromide/D2O under the action of applied viscous torques. Shear forces were applied using a custom-built Couette cell that was introduced into an NMR superconducting magnet, so that its rotational axis was parallel to the magnetic field direction, along which the liquid-crystal director originally aligned. Subsequently, the inner cylinder of the cell was rotated continuously at different rates using a stepper motor. The resulting time evolution and ultimate steady-state orientation of the director, governed by the competition between the applied viscous torque with elastic and magnetic terms, was measured via observed changes in the deuterium spectrum. Using a simple gearbox allowed unprecedented access to a low-shear-rate regime in which, above a threshold shear rate, the director of part of the sample was observed to reorient, while the remaining part still aligned with the magnetic field. Subsequent increases in the applied rotational rate were found to increase the relative proportion of the orienting fraction. Spatially resolved NMR spectra showed that the orienting and field-aligned fractions formed separated bands across the gap of the Couette cell, with director reorientation being initiated at the moving inner wall. The behavior was found to be consistent with the often ignored variation in velocity gradient manifest across the gap of a cylindrical cell, so that as the angular frequency of the inner cylinder was increased the radial location of the critical shear rate required for reorientation traversed the gap. Once the applied rotational rate was sufficient to reorient the director of the entire sample, the dependence of the exhibited steady-state orientation on the average applied shear rate was measured. These results could be fitted to an analytical solution of the force-balance equation, made tractable by the assumption that the elasticity term was of minor significance and could be ignored. Additionally, the use of a numerical solution of the full force-balance equation, which explicitly includes elasticity and secondary flow and additionally allows the time evolution of the director orientation to be calculated, was investigated.

Insights into the potential functionality of single-chain force-induced conformational transitions in polymer networks: implications for polysaccharide signaling in the plant cell wall.

The behavior of biopolymer networks comprised of clickable polysaccharide chains that can undergo force-induced conformational transitions was investigated during straining using a simulation technique. The simulation was carried out both using an affine deformation field and alternatively using Lees-Edwards boundary conditions as an example of a nonaffine case. In the affine situation the simulated stress-strain curves were found to be consistent with results obtained by evaluating the molecular force-extension curve at a single average extension and calculating the bulk modulus as an average over all possible orientations with respect to the deformation. While in all cases examined the macroscopic mechanical responses of networks of randomly oriented chains, consisting either of simple extensible wormlike chains or their clickable analogs, were found to be indistinguishable, the simulation additionally allowed the number of chains containing sugar rings in different conformational states to be monitored, and this was found to change significantly during straining. This supports the hypothesis that in networks of randomly oriented clickable polysaccharide chains, such conformational transitions could have biological significance as stress switches in signaling processes but that they are unlikely to affect the bulk rheological properties of tissue.

Computerized assessment of angiographic occlusion rate and coil density in embolized human cerebral aneurysms.

BACKGROUND AND PURPOSE: Computerized methods have been introduced for more objective quantification of angiographic occlusion rate and coil density as parameters of successful embolization. This study aimed 1) to evaluate this new computerized method for angiographic occlusion rating and coil density calculations by comparison with corresponding histometric parameters from retrieved human aneurysms, and 2) to compare the new computerized method with the present standard of subjective angiographic occlusion rating. MATERIALS AND METHODS: From 14 postmortem-retrieved human aneurysms, angiographic occlusion rate was determined by contrast medium attenuation-gradient distinction on digital subtraction angiographs after Guglielmi detachable coil (GDC) embolization. Angiographic coil density was calculated, approximating aneurysms as ellipsoid and coils as cylindric volumes. On surface-stained histologic ground sections of the respective aneurysms, the occluded aneurysm area and coil area were measured. Then, we calculated and compared the histometric occlusion rates and coil densities with the corresponding angiographic parameters by using the Wilcoxon paired signed-rank test and the Spearman rank correlation. RESULTS: Computerized angiographic occlusion rates (75%-100%) showed good correlation (r = 0.799; P < .01) with histometric occlusion-rates (61%-100%), resulting in no statistically significant differences (P = .2163). With 5.1% (+/-3.8), the mean difference between computerized angiographic occlusion rates and histometry was substantially lower compared with 10.7% (+/-8.7) mean difference between subjective angiographic estimations and histometry. Calculated angiographic coil density (13%-32%) significantly differed from histometric coil density (8%-35%; P < .05). CONCLUSIONS: For recanalized aneurysms, computerized angiographic occlusion rating showed better correspondence with histometry compared with subjective angiographic occlusion rating. Clinical application of this new tool may lead to more objective cutoff values for re-embolization indications. The value of coil density calculations seems limited by the approximation of the aneurysms as ellipsoid volumes.

Adjunctive treatment of acute mania: a clinical overview.

OBJECTIVE: To provide an overview of the available high quality evidence-base of studies of adjunctive pharmacologic treatment for acute mania. METHOD: Double-blind controlled trials with adequate samples (n > 100) were identified through search of PubMed/MEDLINE and computerized abstracts from 2004-2006 meetings of the American Psychiatric Association, International Conference on Bipolar Disorder, and Collegium Internationale Neuro-Psychopharmacolium using key words mania, adjunct, and combination. RESULTS: Placebo-controlled studies with positive results support the adjunctive use of five agents including valproate, olanzapine, risperidone, quetiapine, and haloperidol. Agents with only negative or failed placebo-controlled studies included carbamazepine, gabapentin, lamotrigine, topiramate, oxcarbazepine, and ziprasidone. We found no placebo-controlled study of many commonly used agents including lithium, aripiprazole, and clozapine. No studies explicitly excluded subjects, based on prior treatment with the monotherapy being offered and several studies limited randomization to patients with documented inadequate response to the monotherapy arm. CONCLUSION: The available placebo-controlled randomized clinical trials support adjunctive therapy combining lithium or valproate with olanzapine, risperidone, haloperidol or quetiapine. The additional increment of antimanic efficacy of these combinations over monotherapy was similar in magnitude to that seen for the same agents as monotherapy in comparison with placebo. These additive benefits enhanced the tolerability of adverse effects sufficiently to allow a higher proportion of subjects receiving combination therapies to complete the studies than monotherapy treated patients. The available data has several shortcomings and the available studies are inadequate to conclusively determine whether combination treatment is more efficacious than monotherapy when used by subjects naive to both treatments. Nevertheless, adjunctive treatment, which combines agents with proven antimanic efficacy, offers an attractive option for patients with acute mania.

Effect of a pocket-size tablet-dispensing device on glycaemic control in Type 2 diabetic patients.

AIM: To evaluate whether a pocket-size tablet-dispensing device would improve adherence to therapy as judged by reduction of HbA(1c) levels in a large population of Type 2 diabetic patients. METHODS: The study design was prospective, randomized, open label with two parallel groups. Patients (2296) were recruited from general practitioners and internists and randomized to either receive a tablet dispenser (TD) or no intervention (control group, CO). Patients' characteristics and current oral therapy (including dosage) were recorded at baseline. HbA(1c) was compared between groups at baseline and after 6 months' intervention. RESULTS: Data were available in 2081 patients. Baseline characterisitcs, including age, body mass index (BMI), blood pressure and gender distribution were comparable between the two groups, as was HbA(1c)[7.9 (7.9-8.0), TD vs. 8.0 (7.9-8.0)%, CO, means (95% CI)]. After 6 months, HbA(1c) improved in both groups, but improvement was significantly greater in TD than in controls [-0.74 (0.67-0.80) vs. -0.53 (0.47-0.59)%, P < 0.0001]. Possession of a dispenser remained an independent predictor of improved control in a multiple regression model. In the subgroup analysis, the effect was significantly more pronounced (i) in patients receiving more medications and (ii) more diabetes medications per day (iii) in younger patients. CONCLUSION: In this large study population in a 'realistic' setting, a simple tablet-dispensing device led to a significant and clinically relevant improvement in HbA(1c) levels. Because patients with a more complex therapy regimen benefited more, we suggest that TD might have improved adherence to therapy.

[The influence of conductive hearing loss on DPOAE-threshold. The effect of an individually optimized stimulation]. / Der Einfluss einer Schallleitungsstörung auf die DPOAE-Schwelle. Möglichkeiten einer individuell optimierten Auslösung.

BACKGROUND: The DPOAE-threshold, estimated from extrapolated I/O functions, allows an objective assessment of the mechanical sensitivity of the inner ear. In children, the specificity of this diagnostic tool is impaired by conductive hearing loss. METHODS: In this study, we propose an individual optimization of the primary tone level ratio. This procedure allows the detection of a conductive hearing loss that can be accounted for when estimating the DPOAE-threshold. By means of a simulation using DPOAE-data from 22 normally hearing subjects, the effects of this procedure on the estimation of the DPOAE-threshold are examined. RESULTS: An individually optimized DPOAE stimulation distinctly improves the signal-to-noise ratio of the DPOAE which enables an estimation of the DPOAE-threshold for sound conductive losses up to 15 dB. The DPOAE-threshold only worsens in individual cases. CONCLUSIONS: An individually optimized stimulation paradigm may improve the specificity of inner ear diagnostics with the DPOAE-threshold. A clinical evaluation of the method in children, however, is necessary.