Dreidimensionale Modelle zur Planung komplexer Eingriffe in der kardiovaskulären Chirurgie

Ziel dieser Arbeit war, mit Hilfe von dreidimensionalen, plastischen Modelle komplexe anatomische Situation zu veranschaulichen und die chirurgische Planung zu vereinfachen. Die durchgeführte Routinediagnostik ist im Allgemeinen ausreichend, um kardiale Pathologien exakt zu veranschaulichen, Diagnosen zu stellen und Behandlungsstrategien zu entwerfen. Bei sehr komplexen anatomischen Situationen mit vaskulären und kardialen Fehlbildungen und zusätzlicher Narbenbildung ist es jedoch mitunter schwierig die kardiale Anatomie vollständig zu verstehen und die relevanten Strukturen zu identifizieren. Unter diesen Umständen kann eine dreidimensionale Darstellung hilfreich sein. Zusätzlich zu virtuellen 3D-Rekonstruktionen, die lediglich am Computer betrachtet werden können, bieten plastische Modelle die Möglichkeit, Eingriffe zu simulieren, Devices zu testen und die Modelle zur intraoperativen Orientierung mit in den Operationssaal zu nehmen. Fortschritte in der medizinischen Bildgebung und der Bildbearbeitungssoftware haben die Anwendung von anatomischen Modellbau-Verfahren in der Herzchirurgie ermöglicht. Basierend auf routinemäßig erstellter Diagnostik, wie CT, MRT und MR-Angiographie gelang es für unterschiedliche Indikationen in der Herzchirurgie mittels spezieller Software und dem 3D-Printing Verfahren dreidimensionale Modelle zu erstellen. Die in dieser Arbeit realisierten Indikationen umfassen Pathologien aus den Bereichen der Kinderherzchirurgie, Transplantationschirurgie, Erwachsenenherzchirurgie und der interventionellen Kardiologie. Die Modelle wurden zur präoperativen Planung und intraoperativen Orientierung im Operationssaal eingesetzt. Anhand der Modelle konnten maßgeschneiderte Devices für die interventionelle Kardiologie entwickelt werden und deren Einbringen präoperativ getestet werden. Es hat sich gezeigt, dass die Anwendung von plastischen Modellen zusätzlich zur Routinediagnostik in komplexen Fällen gerechtfertigt ist und dem behandelnden Team die Operationsplanung und die Orientierung im Situs vereinfachen kann

Bacterial expression of mutant argininosuccinate lyase reveals imperfect correlation of in - vitro enzyme activity with clinical phenotype in argininosuccinic aciduria

Background: The urea cycle defect argininosuccinate lyase (ASL) deficiency has a large spectrum of presentations from highly severe to asymptomatic. Enzyme activity assays in red blood cells or fibroblasts, although diagnostic of the deficiency, fail to discriminate between severe, mild or asymptomatic cases. Mutation/phenotype correlation studies are needed to characterize the effects of individual mutations on the activity of the enzyme. Methods: Bacterial in-vitro expression studies allowed the enzyme analysis of purified mutant ASL proteins p.I100T (c.299T > C), p.V178M (c.532G > A), p.E189G (c.566A > G), p.Q286R (c.857A > G), p.K315E (c.943A > G), p.R379C (c.1135C > T) and p.R385C (c.1153C > T) in comparison to the wildtype protein. Results: In the bacterial in-vitro expression system, ASL wild-type protein was successfully expressed. The known classical p.Q286R, the novel classical p.K315E and the known mutations p.I100T, p.E189G and p.R385C, which all have been linked to a mild phenotype, showed no significant residual activity. There was some enzyme activity detected with the p.V178M (5 % of wild-type) and p.R379C (10 % of wild-type) mutations in which Km values for argininosuccinic acid differed significantly from the wild-type ASL protein. Conclusion: The bacterially expressed enzymes proved that the mutations found in patients and studied here indeed are detrimental. However, as in the case of red cell ASL activity assays, some mutations found in genetically homozygous patients with mild presentations resulted in virtual loss of enzyme activity in the bacterial system, suggesting a more protective environment for the mutant enzyme in the liver than in the heterologous expression system and/or in the highly dilute assays utilized her

The well-preserved Late Neolithic dolmen burial of Oberbipp, Switzerland. Construction, use, and post-depositional processes

Excavation of the Late Neolithic dolmen of Oberbipp BE, Steingasse in the Swiss Central Plateau provided a unique opportunity for a comprehensive study of the archaeological and anthropological evidence. In multidisciplinary studies, we investigated the processes at work during construction, use, and abandonment of the megalithic structure, as well as the dietary habits, subsistence strategy, and possible mobility of the Neolithic population. Archaeological methods included micromorphology, archaeobiology, typology, use-wear analysis, and geology. The anthropological investigation was complemented by an analysis of stable isotope ratios and palaeogenetics. Local topography and the cover of alluvial sediments ensured an extraordinary conservation of the monument. It allowed the preservation of the human remains of at least 42 individuals of both sexes and all ages. The observation of the sedimentary and post-depositional processes, supplemented by an extensive series of radiocarbon dates, allowed us to reconstruct the history of the dolmen in its environment and the definition of at least two deposition phases. We found genetic evidence of lactase intolerance, a local population with a mixed ancestry of early Anatolian farmers and Western hunter-gatherers, and a crop-based diet. Sparse remains of a nearby Late Neolithic settlement sustain the interpretation that this is the burial site of a local farming community. Evidence of higher mobility of females and kinship over three generations solely in the paternal line suggests a virilocal community. Bone-altering pathologies support the assumption of a caring society

Unstable argininosuccinate lyase in variant forms of the urea cycle disorder argininosuccinic aciduria

Loss of function of the urea cycle enzyme argininosuccinate lyase (ASL) is caused by mutations in the ASL gene leading to ASL deficiency (ASLD). ASLD has a broad clinical spectrum ranging from life-threatening severe neonatal to asymptomatic forms. Different levels of residual ASL activity probably contribute to the phenotypic variability but reliable expression systems allowing clinically useful conclusions are not yet available. In order to define the molecular characteristics underlying the phenotypic variability, we investigated all ASL mutations that were hitherto identified in patients with late onset or mild clinical and biochemical courses by ASL expression in human embryonic kidney 293T cells. We found residual activities >3% of ASL wild type (WT) in nine of 11 ASL mutations. Six ASL mutations (p.Arg95Cys, p.Ile100Thr, p.Val178Met, p.Glu189Gly, p.Val335Leu, and p.Arg379Cys) with residual activities ≥16% of ASL WT showed no significant or less than twofold reduced Km values, but displayed thermal instability. Computational structural analysis supported the biochemical findings by revealing multiple effects including protein instability, disruption of ionic interactions and hydrogen bonds between residues in the monomeric form of the protein, and disruption of contacts between adjacent monomeric units in the ASL tetramer. These findings suggest that the clinical and biochemical course in variant forms of ASLD is associated with relevant residual levels of ASL activity as well as instability of mutant ASL proteins. Since about 30% of known ASLD genotypes are affected by mutations studied here, ASLD should be considered as a candidate for chaperone treatment to improve mutant protein stability

Unstable argininosuccinate lyase in variant forms of the urea cycle disorder argininosuccinic aciduria.

Loss of function of the urea cycle enzyme argininosuccinate lyase (ASL) is caused by mutations in the ASL gene leading to ASL deficiency (ASLD). ASLD has a broad clinical spectrum ranging from life-threatening severe neonatal to asymptomatic forms. Different levels of residual ASL activity probably contribute to the phenotypic variability but reliable expression systems allowing clinically useful conclusions are not yet available. In order to define the molecular characteristics underlying the phenotypic variability, we investigated all ASL mutations that were hitherto identified in patients with late onset or mild clinical and biochemical courses by ASL expression in human embryonic kidney 293 T cells. We found residual activities >3 % of ASL wild type (WT) in nine of 11 ASL mutations. Six ASL mutations (p.Arg95Cys, p.Ile100Thr, p.Val178Met, p.Glu189Gly, p.Val335Leu, and p.Arg379Cys) with residual activities ≥16 % of ASL WT showed no significant or less than twofold reduced Km values, but displayed thermal instability. Computational structural analysis supported the biochemical findings by revealing multiple effects including protein instability, disruption of ionic interactions and hydrogen bonds between residues in the monomeric form of the protein, and disruption of contacts between adjacent monomeric units in the ASL tetramer. These findings suggest that the clinical and biochemical course in variant forms of ASLD is associated with relevant residual levels of ASL activity as well as instability of mutant ASL proteins. Since about 30 % of known ASLD genotypes are affected by mutations studied here, ASLD should be considered as a candidate for chaperone treatment to improve mutant protein stability

Whole-body CT in haemodynamically unstable severely injured patients--a retrospective, multicentre study.

The current common and dogmatic opinion is that whole-body computed tomography (WBCT) should not be performed in major trauma patients in shock. We aimed to assess whether WBCT during trauma-room treatment has any effect on the mortality of severely injured patients in shock.In a retrospective multicenter cohort study involving 16719 adult blunt major trauma patients we compared the survival of patients who were in moderate, severe or no shock (systolic blood pressure 90-110,<90 or >110 mmHg) at hospital admission and who received WBCT during resuscitation to those who did not. Using data derived from the 2002-2009 version of TraumaRegister®, we determined the observed and predicted mortality and calculated the standardized mortality ratio (SMR) as well as logistic regressions.9233 (55.2%) of the 16719 patients received WBCT. The mean injury severity score was 28.8±12.1. The overall mortality rate was 17.4% (SMR  = 0.85, 95%CI 0.81-0.89) for patients with WBCT and 21.4% (SMR = 0.98, 95%CI 0.94-1.02) for those without WBCT (p<0.001). 4280 (25.6%) patients were in moderate shock and 1821 (10.9%) in severe shock. The mortality rate for patients in moderate shock with WBCT was 18.1% (SMR 0.85, CI95% 0.78-0.93) compared to 22.6% (SMR 1.03, CI95% 0.94-1.12) to those without WBCT (p<0.001, p = 0.002 for the SMRs). The mortality rate for patients in severe shock with WBCT was 42.1% (SMR 0.99, CI95% 0.92-1.06) compared to 54.9% (SMR 1.10, CI95% 1.02-1.16) to those without WBCT (p<0.001, p = 0.049 for the SMRs). Adjusted logistic regression analyses showed that WBCT is an independent predictor for survival that significantly increases the chance of survival in patients in moderate shock (OR = 0.73; 95%CI 0.60-0.90, p = 0.002) as well as in severe shock (OR = 0.67; 95%CI 0.52-0.88, p = 0.004). The number needed to scan related to survival was 35 for all patients, 26 for those in moderate shock and 20 for those in severe shock.WBCT during trauma resuscitation significantly increased the survival in haemodynamically stable as well as in haemodynamically unstable major trauma patients. Thus, the application of WBCT in haemodynamically unstable severely injured patients seems to be safe, feasible and justified if performed quickly within a well-structured environment and by a well-organized trauma team

Bacterial expression of mutant argininosuccinate lyase reveals imperfect correlation of in-vitro enzyme activity with clinical phenotype in argininosuccinic aciduria

BACKGROUND: The urea cycle defect argininosuccinate lyase (ASL) deficiency has a large spectrum of presentations from highly severe to asymptomatic. Enzyme activity assays in red blood cells or fibroblasts, although diagnostic of the deficiency, fail to discriminate between severe, mild or asymptomatic cases. Mutation/phenotype correlation studies are needed to characterize the effects of individual mutations on the activity of the enzyme. METHODS: Bacterial in-vitro expression studies allowed the enzyme analysis of purified mutant ASL proteins p.I100T (c.299 T > C), p.V178M (c.532 G > A), p.E189G (c.566A > G), p.Q286R (c.857A > G), p.K315E (c.943A > G), p.R379C (c.1135 C > T) and p.R385C (c.1153 C > T) in comparison to the wildtype protein. RESULTS: In the bacterial in-vitro expression system, ASL wild-type protein was successfully expressed. The known classical p.Q286R, the novel classical p.K315E and the known mutations p.I100T, p.E189G and p.R385C, which all have been linked to a mild phenotype, showed no significant residual activity. There was some enzyme activity detected with the p.V178M (5 % of wild-type) and p.R379C (10 % of wild-type) mutations in which K(m) values for argininosuccinic acid differed significantly from the wild-type ASL protein. CONCLUSION: The bacterially expressed enzymes proved that the mutations found in patients and studied here indeed are detrimental. However, as in the case of red cell ASL activity assays, some mutations found in genetically homozygous patients with mild presentations resulted in virtual loss of enzyme activity in the bacterial system, suggesting a more protective environment for the mutant enzyme in the liver than in the heterologous expression system and/or in the highly dilute assays utilized here

The Revised Injury Severity Classification (RISC) – Score.

<p>New ISS  =  new injury severity score; GCS  =  Glasgow coma scale; PTT  =  partial thromboplastin time; * Systolic blood pressure <90 mmHg/haemoglobin level <9 g/dL/≥10 units of packed red blood cells; The RISC score is used to calculate the probability of death of a trauma patient. It is one of the most precise prognostic major trauma score <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068880#pone.0068880-Lefering1" target="_blank">[28]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068880#pone.0068880-Lefering2" target="_blank">[44]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068880#pone.0068880-Ruchholtz2" target="_blank">[45]</a>.</p

Characteristics of severely injured patients with information about CT during trauma room treatment.

<p>Data are given as number of patients (% of total patients) or mean ± SD, unless indicated otherwise. SBP Systolic blood pressure, GCS Glasgow Coma Scale; ICU Intensive Care Unit; PRBC packed red blood cells; ISS Injury Severity Score; AIS abbreviated injury scale. RISC  =  revised injury severity classification score; p-value (comparison of WBCT vs. non-WBCT group): χ² test or Mann-Whitney-U test (two sided), *MOF, defined as organ failure of two systems of >2 SOFA-score points of at least 2 days duration <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068880#pone.0068880-Vincent1" target="_blank">[46]</a>.</p