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Sussulini A.,University of Campinas | Sussulini A.,Julich Research Center | Sussulini A.,Max Planck Institute for Experimental Medicine | Kratzin H.,Max Planck Institute for Experimental Medicine | And 5 more authors.
Analytical Chemistry | Year: 2010

In the present work, metallomics studies using biomolecular (matrix-assisted laser desorption ionization time-of-flight tandem mass spectrometry, MALDI-TOF MS/MS) and elemental mass spectrometry (laser ablation inductively coupled plasma mass spectrometry, LA-ICPMS) of human blood serum samples from bipolar disorder (BD) patients compared to controls were performed. The serum samples from three different groups: control (n = 25), BD patients treated with Li (n = 15), and BD patients not treated with Li (n = 10), were pooled according to their groups and separated by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). Then, in order to determine the metals bound to the protein spots and search for differences among the studied groups, the 2-D gels were analyzed by LA-ICPMS in three distinct modes: bioimaging of metals in gel sections, line scan through the protein spots, and microlocal analysis of selected protein spots. MALDI-TOF MS/MS characterized 32 serum proteins, and they were associated with the metals previously detected. When comparing control and treated BD patient groups, a differentiation in terms of metals bound to proteins was possible to observe. The main metals bound to proteins found in all groups were Na, Mg, Zn, Ca, and Fe. Mn was only detected in the control group; Co was only observed in the control and BD patients treated with Li group. K and Ti were only found in the BD patient groups, and P was only observed in control and BD patients not treated with Li drugs. This exploratory work shows that the association of LA-ICPMS with MALDI-TOF MS/MS is a powerful strategy in metallomics studies applied to determine differences in metal-containing proteins, being able to play an important role on the discovery of potential markers for BD and its treatment with Li in serum samples. © 2010 American Chemical Society.

Wessels J.T.,University of Gottingen | Yamauchi K.,Anticancer, Inc. | Yamauchi K.,University of California at San Diego | Hoffman R.M.,Anticancer, Inc. | And 3 more authors.
Cytometry Part A | Year: 2010

This review focuses on technical advances in fluorescence microscopy techniques including laser scanning techniques, fluorescence-resonance energy transfer (FRET) microscopy, fluorescence lifetime imaging (FLIM), stimulated emission depletion (STED)-based super-resolution microscopy, scanning confocal endomicroscopes, thinsheet laser imaging microscopy (TSLIM), and tomographic techniques such as early photon tomography (EPT) as well as on clinical laser-based endoscopic and microscopic techniques. We will also discuss the new developments in the field of fluorescent dyes and fluorescent genetic reporters that enable new possibilities in high-resolution and molecular imaging both in in vitro and in vivo. Small animal and tissue imaging benefit from the development of new fluorescent proteins, dyes, and sensing constructs that operate in the far red and near-infrared spectrum. Copyright © 2010 International Society for Advancement of Cytometry.

Pajonk F.-G.,Saarland University | Pajonk F.-G.,Center for Psychiatric and Psychotherapeutic Care and Rehabilitation | Wobrock T.,University of Gottingen | Gruber O.,University of Gottingen | And 13 more authors.
Archives of General Psychiatry | Year: 2010

Context: Hippocampal volume is lower than expected in patients with schizophrenia; however, whether this represents a fixed deficit is uncertain. Exercise is a stimulus to hippocampal plasticity. Objective: To determine whether hippocampal volume would increase with exercise in humans and whether this effect would be related to improved aerobic fitness. Design: Randomized controlled study. Setting: Patients attending a day hospital program or an outpatient clinic. Patients or Other Participants: Male patients with chronic schizophrenia and matched healthy subjects. Interventions: Aerobic exercise training (cycling) and playing table football (control group) for a period of 3 months. Main Outcome Measures: Magnetic resonance imaging of the hippocampus. Secondary outcome measures were magnetic resonance spectroscopy, neuropsychological (Rey Auditory Verbal Learning Test, Corsi blocktapping test), and clinical (Positive and Negative Syndrome Scale) features. Results: Following exercise training, relative hippocampal volume increased significantly in patients (12%) and healthy subjects (16%), with no change in the nonexercise group of patients (-1%). Changes in hippocampal volume in the exercise group were correlated with improvements in aerobic fitness measured by change in maximum oxygen consumption (r=0.71; P=.003). In the schizophrenia exercise group (but not the controls), change in hippocampal volume was associated with a 35% increase in the N-acetylaspartate to creatine ratio in the hippocampus. Finally, improvement in test scores for short-term memory in the combined exercise and nonexercise schizophrenia group was correlated with change in hippocampal volume (r=0.51; P<.05). Conclusion: These results indicate that in both healthy subjects and patients with schizophrenia hippocampal volume is plastic in response to aerobic exercise. ©2010 American Medical Association. All rights reserved.

Bonn F.,University of Cologne | Pantakani K.,University of Gottingen | Shoukier M.,University of Gottingen | Langer T.,University of Cologne | And 3 more authors.
Human Mutation | Year: 2010

An autosomal recessive form of hereditary spastic paraplegia (AR-HSP) is primarily caused by mutations in the SPG7 gene, which codes for paraplegin, a subunit of the hetero-oligomeric m-AAA protease in mitochondria. In the current study, sequencing of the SPG7 gene in the genomic DNA of 25 unrelated HSP individuals/families led to the identification of two HSP patients with compound heterozygous mutations (p.G349S/p.W583C and p.A510V/p.N739KfsX741) in the coding sequence of the SPG7 gene. We used a yeast complementation assay to evaluate the functional consequence of novel SPG7 sequence variants detected in the HSP patients. We assessed the proteolytic activity of hetero-oligomeric m-AAA proteases composed of paraplegin variant(s) and proteolytically inactive forms of AFG3L2 (AFG3L2E575Q or AFG3L2K354A) upon expression in m-AAA protease-deficient yeast cells. We demonstrate that the newly identified paraplegin variants perturb the proteolytic function of hetero-oligomeric m-AAA protease. Moreover, commonly occurring silent polymorphisms such as p.T503A and p.R688Q could be distinguished from mutations (p.G349S, p.W583C, p.A510V, and p.N739KfsX741) in our HSP cohort. The yeast complementation assay thus can serve as a reliable system to distinguish a pathogenic mutation from a silent polymorphism for any novel SPG7 sequence variant, which will facilitate the interpretation of genetic data for SPG7. © 2010 Wiley-Liss, Inc.

Ducker E.B.,Max Planck Institute for Biophysical Chemistry | Kuhn L.T.,EXC 171 | Kuhn L.T.,European Neuroscience Institute Gottingen ENI G | Munnemann K.,Max Planck Institute for Polymer Research | And 2 more authors.
Journal of Magnetic Resonance | Year: 2012

The Non-Hydrogenative Parahydrogen-Induced Polarization (NH-PHIP) technique, which is referred to as Signal Amplification by Reversible Exchange (SABRE), has been reported to be applicable to various substrates and catalysts. For more detailed studies, pyridine was mainly examined in the past. Here, we examined several pyrazole derivatives towards their amenability to this method using Crabtree's Catalyst, which is the polarization transfer catalyst that is best documented. Additionally, the dependence of the signal enhancement on the field strength, at which the polarization step takes place, was examined for pyridine and four different pyrazoles. To achieve this, the polarization step was performed at numerous previously determined magnetic fields in the stray field of the NMR spectrometer. The substrate dependence of the field dependence proved to be relatively small for the different pyrazoles and a strong correlation to the field dependence for pyridine was observed. Reducing the number of spins in the catalyst by deuteration leads to increased enhancement. This indicates that more work has to be invested in order to be able to reproduce the SABRE field dependence by simulations. © 2011 Elsevier Inc. All rights reserved.

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