In Situ Monitoring of the Catalytic Activity of Cytochrome c Oxidase in a Biomimetic Architecture

AbstractCytochrome c oxidase (CcO) from Paracoccus denitrificans was immobilized in a strict orientation via a his-tag attached to subunit I on a gold film and reconstituted in situ into a protein-tethered bilayer lipid membrane. In this orientation, the cytochrome c (cyt c) binding site is directed away from the electrode pointing to the outer side of the protein-tethered bilayer lipid membrane architecture. The CcO can thus be activated by cyt c under aerobic conditions. Catalytic activity was monitored by impedance spectroscopy, as well as cyclic voltammetry. Cathodic and anodic currents of the CcO with cyt c added to the bulk solution were shown to increase under aerobic compared to anaerobic conditions. Catalytic activity was considered in terms of repeated electrochemical oxidation/reduction of the CcO/cyt c complex in the presence of oxygen. The communication of cyt c bound to the CcO with the electrode is discussed in terms of a hopping mechanism through the redox sites of the enzyme. Simulations supporting this hypothesis are included

Ecstasykonsumenten: Neurokognitive und funktionelle Problemkonstellationen und Ansätze zu spezifischen Frühinterventionen

Hintergrund: In den letzten Jahren ist zunehmend deutlich geworden, dass Konsumenten von Ecstasy sich hinsichtlich Gebrauchsmuster und -kontext wie auch Spontanverlauf, Risiken und Konsequenzen von Konsumenten anderer legaler und illegaler Substanzen unterscheiden und möglicherweise eine recht eigenständige Gruppe darstellen. Diese eigenständige Gruppe wird im angelsächsischen Raum zum Teil auch als club drug user bezeichnet. Alarmierend waren die Vermutungen aus Voruntersuchungen, dass club drug-Konsumenten auch nach dem Konsum geringer Mengen von Ecstasy bemerkenswerte und möglicherweise überdauernde neurobiologische Veränderungen mit assoziierten kognitiven Beeinträchtigungen und Störungen aufzeigen. Dies stellt an sich eine mögliche Gefährdung der Konsumenten dar, zusätzlich wiederum können kognitive Veränderungen auch Einfluss nehmen auf den Verlauf des weiteren Suchtmittelkonsums und den Erfolg von Interventionen. Ziel: In der MAYA-Studie (Munich Assessment of Young Adults) werden an einer epidemiologischen Bevölkerungsstichprobe junger Erwachsener (Stichprobe A) sowie an einer klinischen Stichprobe von Ecstasy-Konsumenten (Stichprobe B) die Art und das Ausmaß kognitiver Störungen und Defizite in Abhängigkeit von Gebrauchsmustern und anderen Einflussfaktoren untersucht. Bei der Stichprobe A handelt es sich um ein Subsample der EDSPStudie. Zusätzlich zu den bereits erhobenen Charakterisierungen werden spezifische neurokognitive Maße (vor allem Aufmerksamkeit, Gedächtnis und exekutive Funktionen) und Fragebögen (Impulsivität, BDI, STAI etc.) erhoben. Die Probanden erhalten weiterhin ein Screening mit dem neu eingeführten Instrument WHO ASSIST (Alcohol, Smoking and Substance Involvement Screening Test). Wenn indiziert, wird eine Intervention im Sinne eines Motivational Enhancement durchgeführt. Initiale Auswirkungen werden in einem Telefoninterview sechs Wochen später überprüft. Ergebnisse: Die vorläufigen Ergebnisse beruhen auf einer Teilstichprobe. Insgesamt handelte es sich eher um Konsumenten mit geringgradigem bis moderaten Konsum. Dennoch ließen sich Unterschiede zwischen den Konsumentengruppen (Ecstasy, Cannabis, Alkohol) und den Nichtkonsumenten erkennen. Die Konsumenten von Ecstasy unterschieden sich am ausgeprägtesten von den Kontrollen. Die sich bisher abzeichnenden Unterschiede bei Aufmerksamkeit, Gedächtnis und exekutiven Funktionen scheinen bei der Gedächtnis- und Merkfähigkeit am ausgeprägtesten zu sein. Bei diesen vorläufigen Ergebnissen sind die Untersuchungsgruppen noch sehr heterogen und enthalten z.B. Probanden, die ihren Ecstasykonsum bereits wieder aufgegeben haben.Introduction: In recent years it has become increasingly evident that ecstasy users represent a group distinct from users of other drugs. This is based on consumption patterns, context of use, development of use patterns and other factors. This group of users might be considered "club drug users", given the overlap, similarity of use patterns and consumptions within the class of club drugs. In recent neurobiological research, alarming results have been reported, indicating that persistent neurotoxic effects with concomitant cognitive problems may be induced by ecstasy consumption. Methods: In the Munich Assessment of Young Adults Study (MAYA) two samples of ecstasy users are investigated. Sample A is a sample of users drawn from an ongoing epidemiological longitudinal study of young adults in Munich. Sample B is an additional clinical sample. Both samples are characterized extensively and are neuropsychologically tested for attention, memory and executive functioning. A screening for harmful use is conducted with the German version of the WHO ASSIST (Alcohol, Smoking and Substance Involvement Screening Test). A Motivational Enhancement Intervention is applied when harmful use is detected. With a telephone interview the intervention's initial effect is assessed. Results: Initial results are based on a small sample. Use within the tested population is low to moderate. Still significant differences in attention, memory, and executive functioning could be detected with the current testing. Ecstasy users almost consistently showed the lowest cognitive functioning. The one difference is that ecstasy users were significantly faster in finger tapping and reaction. Note that the results are preliminary. The sample is heterogeneous, including both current and former users. Firmer results will be reported with the full sample size, allowing to further elucidate subgroups and interactions

Glucose transporter-1 deficiency syndrome: the expanding clinical and genetic spectrum of a treatable disorder

Glucose transporter-1 deficiency syndrome is caused by mutations in the SLC2A1 gene in the majority of patients and results in impaired glucose transport into the brain. From 2004-2008, 132 requests for mutational analysis of the SLC2A1 gene were studied by automated Sanger sequencing and multiplex ligation-dependent probe amplification. Mutations in the SLC2A1 gene were detected in 54 patients (41%) and subsequently in three clinically affected family members. In these 57 patients we identified 49 different mutations, including six multiple exon deletions, six known mutations and 37 novel mutations (13 missense, five nonsense, 13 frame shift, four splice site and two translation initiation mutations). Clinical data were retrospectively collected from referring physicians by means of a questionnaire. Three different phenotypes were recognized: (i) the classical phenotype (84%), subdivided into early-onset (<2 years) (65%) and late-onset (18%); (ii) a non-classical phenotype, with mental retardation and movement disorder, without epilepsy (15%); and (iii) one adult case of glucose transporter-1 deficiency syndrome with minimal symptoms. Recognizing glucose transporter-1 deficiency syndrome is important, since a ketogenic diet was effective in most of the patients with epilepsy (86%) and also reduced movement disorders in 48% of the patients with a classical phenotype and 71% of the patients with a non-classical phenotype. The average delay in diagnosing classical glucose transporter-1 deficiency syndrome was 6.6 years (range 1 month-16 years). Cerebrospinal fluid glucose was below 2.5 mmol/l (range 0.9-2.4 mmol/l) in all patients and cerebrospinal fluid : blood glucose ratio was below 0.50 in all but one patient (range 0.19-0.52). Cerebrospinal fluid lactate was low to normal in all patients. Our relatively large series of 57 patients with glucose transporter-1 deficiency syndrome allowed us to identify correlations between genotype, phenotype and biochemical data. Type of mutation was related to the severity of mental retardation and the presence of complex movement disorders. Cerebrospinal fluid : blood glucose ratio was related to type of mutation and phenotype. In conclusion, a substantial number of the patients with glucose transporter-1 deficiency syndrome do not have epilepsy. Our study demonstrates that a lumbar puncture provides the diagnostic clue to glucose transporter-1 deficiency syndrome and can thereby dramatically reduce diagnostic delay to allow early start of the ketogenic die

MedShapeNet -- A Large-Scale Dataset of 3D Medical Shapes for Computer Vision

Prior to the deep learning era, shape was commonly used to describe the objects. Nowadays, state-of-the-art (SOTA) algorithms in medical imaging are predominantly diverging from computer vision, where voxel grids, meshes, point clouds, and implicit surface models are used. This is seen from numerous shape-related publications in premier vision conferences as well as the growing popularity of ShapeNet (about 51,300 models) and Princeton ModelNet (127,915 models). For the medical domain, we present a large collection of anatomical shapes (e.g., bones, organs, vessels) and 3D models of surgical instrument, called MedShapeNet, created to facilitate the translation of data-driven vision algorithms to medical applications and to adapt SOTA vision algorithms to medical problems. As a unique feature, we directly model the majority of shapes on the imaging data of real patients. As of today, MedShapeNet includes 23 dataset with more than 100,000 shapes that are paired with annotations (ground truth). Our data is freely accessible via a web interface and a Python application programming interface (API) and can be used for discriminative, reconstructive, and variational benchmarks as well as various applications in virtual, augmented, or mixed reality, and 3D printing. Exemplary, we present use cases in the fields of classification of brain tumors, facial and skull reconstructions, multi-class anatomy completion, education, and 3D printing. In future, we will extend the data and improve the interfaces. The project pages are: https://medshapenet.ikim.nrw/ and https://github.com/Jianningli/medshapenet-feedbackComment: 16 page

A principal component meta-analysis on multiple anthropometric traits identifies novel loci for body shape

Large consortia have revealed hundreds of genetic loci associated with anthropometric traits, one trait at a time. We examined whether genetic variants affect body shape as a composite phenotype that is represented by a combination of anthropometric traits. We developed an approach that calculates averaged PCs (AvPCs) representing body shape derived from six anthropometric traits (body mass index, height, weight, waist and hip circumference, waist-to-hip ratio). The first four AvPCs explain >99% of the variability, are heritable, and associate with cardiometabolic outcomes. We performed genome-wide association analyses for each body shape composite phenotype across 65 studies and meta-analysed summary statistics. We identify six novel loci: LEMD2 and CD47 for AvPC1, RPS6KA5/C14orf159 and GANAB for AvPC3, and ARL15 and ANP32 for AvPC4. Our findings highlight the value of using multiple traits to define complex phenotypes for discovery, which are not captured by single-trait analyses, and may shed light onto new pathways

New genetic loci link adipose and insulin biology to body fat distribution.

Body fat distribution is a heritable trait and a well-established predictor of adverse metabolic outcomes, independent of overall adiposity. To increase our understanding of the genetic basis of body fat distribution and its molecular links to cardiometabolic traits, here we conduct genome-wide association meta-analyses of traits related to waist and hip circumferences in up to 224,459 individuals. We identify 49 loci (33 new) associated with waist-to-hip ratio adjusted for body mass index (BMI), and an additional 19 loci newly associated with related waist and hip circumference measures (P < 5 × 10(-8)). In total, 20 of the 49 waist-to-hip ratio adjusted for BMI loci show significant sexual dimorphism, 19 of which display a stronger effect in women. The identified loci were enriched for genes expressed in adipose tissue and for putative regulatory elements in adipocytes. Pathway analyses implicated adipogenesis, angiogenesis, transcriptional regulation and insulin resistance as processes affecting fat distribution, providing insight into potential pathophysiological mechanisms

Abstracts from the 8th International Conference on cGMP Generators, Effectors and Therapeutic Implications

This work was supported by a restricted research grant of Bayer AG