Subregions of the ventral striatum show preferential coding of reward magnitude and probability

As shown in non-human primate and human fMRI studies the probability and magnitude of anticipated rewards modulate activity in the mesolimbic dopaminergic system. Importantly, non-human primate data have revealed that single dopaminergic neurons code for both probability and magnitude of expected reward, suggesting an identical system. Using a guessing task that allowed the independent assessment of the factors probability and magnitude we were able to assess the impact of reward probability and magnitude in ventral striatal subregions in a large sample (n=98). We observed more anterior and lateral peak activation foci in the ventral striatum for reward probability and a more posterior and medial activation peak for reward magnitude, suggesting a functional segregation at the mesoscopic level. Importantly, this functional bias observed for the group average was also tested in each individual subject, allowing for proper random effects inference for the spatial dissociation. Taken together, our data point toward a functional dissociation of neuronal assemblies suggesting that certain populations of neurons are more sensitive to expected reward probability and other populations are more sensitive to reward magnitude.     

In vivo selection and chemoprotection after drug resistance gene therapy in a nonmyeloablative allogeneic transplantation setting in dogs

We have previously demonstrated successful in vivo selection, chemoprotection, and modulation of donor chimerism in dogs that received myeloablative allogeneic stem cell transplantation with cells expressing the P140K mutant of the DNA repair enzyme methylguanine methyltransferase (MGMTP140K). Here, we wished to investigate whether in vivo selection, chemoprotection, and modulation of donor chimerism could also be achieved after nonmyeloablative transplantation, which could allow for less toxic transplantation regimens for patients with malignant and genetic diseases. Three dogs received a nonmyeloablative conditioning regimen and infusion of allogeneic stem cells transduced with MGMTP140K. All three dogs had stable gene marking and donor chimerism before receiving a course of O(6) -benzylguanine (O(6) BG)/1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) between days 210 and 589 after transplantation. One to four doses led to a marked increase in gene marking in all dogs. Furthermore, the transduced cells conferred chemoprotection and prevented severe neutropenia. Our results suggest that drug resistance gene therapy is feasible and safe in the nonmyeloablative transplantation setting.     

Magnetic resonance imaging of carotid atherosclerosis: plaque analysis

OBJECTIVES: The Computer-Aided System for CArdiovascular Disease Evaluation (CASCADE) has been developed for streamlined, automated analysis of carotid artery magnetic resonance imaging to measure atherosclerotic plaque burden and composition in vivo. The purpose of this investigation was to assess the performance of CASCADE compared with manual outlining. METHODS: Magnetic resonance images were obtained from 26 subjects with 16% to 79% carotid artery stenosis by duplex ultrasound who were imaged twice in a 2-week period with a multiple-slice, multiple-contrast magnetic resonance imaging protocol as part of the Outcome of Rosuvastatin treatment on carotid artery atheroma: a magnetic resonance Imaging ObservatioN trial. Manual outlining was used to identify the boundaries of the lumen, wall, necrotic core (NC), and calcifications. After 6 months, the analysis was repeated using CASCADE. For each data set, the contours were used to compute the maximal normalized wall index (NWI; wall area divided by total vessel area), maximal wall thickness (WT), and the average NC and calcified (CA) areas per slice. Agreement between manual and automated reviews and the scan-scan measurement reproducibilities were evaluated. RESULTS: Pearson correlation between manual and automated analyses was 0.94 for maximal NWI, 0.86 for maximal WT, 0.84 for NC, and 0.96 for CA. Intraclass correlation coefficients for manual and automated analyses were 0.90 and 0.97 for maximal NWI, 0.89 and 0.95 for maximal WT, 0.95 and 0.87 for NC, and 0.96 and 0.94 for CA, respectively. CONCLUSIONS: Automated analysis tools are capable of providing accurate and reproducible measurements of carotid atherosclerotic burden and composition when compared with manually outlined results.     

Serotoninergic receptors on human airway epithelial cells

There is accumulating evidence that points to a role of serotonin (5-hydroxytryptamine [5-HT]) in the pathophysiology of asthma. Therefore, we analyzed the expression of serotoninergic receptors (5-HTR), its linkage to intracellular calcium homeostasis, and its influence on the production and secretion of IL-6, prostaglandin E(2), the CCL-Chemokine CCL5/Rantes, and the CXC-chemokines CXCL8/IL-8, CXCL9/MIG, CXCL10/IP-10, and CXCL11/I-TAC in primary alveolar epithelial cells type II and the human lung cell lines A549 and BEAS-2B. Employing a PCR approach we were able to demonstrate mRNA expression of several 5-HTR, such as the heptahelical receptors 5-HTR1A, 5-HTR1B, 5-HTR1E, 5-HTR1F, 5-HTR2A, 5-HTR4, 5-HTR6, and 5-HTR7, as well as the ligand-gated ion channel 5-HTR3 in alveolar epithelial cells type II (AEC-II), A549, and BEAS-2B cells. To verify functional expression of 5-HTR subtypes, Ca(2+)-transients were analyzed. This enabled us to show that 5-HT induced an increase in intracellular calcium. Further experiments with isotype-selective receptor agonists allowed us to demonstrate that 5-HT induced calcium transients via activation of 5-HTR1, 5-HTR2, and 5-HTR3 in A549 and BEAS-2B cells. Moreover, we revealed that stimulation of 5-HTR1 and 5-HTR2 induced Ca(2+) mobilization from intracellular stores, whereas activation of 5-HTR3 induced Ca(2+) influx from the extracellular space. Functional studies indicated that activation of 5-HTR1B, 5-HTR1E/F, 5-HTR2, 5-HTR3, 5-HTR4, and 5-HTR7 regulated the release of the cytokine IL-6 and the CXC-chemokine CXCL8/IL-8. Our study shows that 5-HT stimulates different signaling pathways and regulates cytokine release in airway epithelial cells. In summary, our data implicate a pathophysiologic role of 5-HT in the asthmatic inflammatory responses in human airway epithelial cells.     

Maturation of synaptic contacts in differentiating neural stem cells

NSCs are found in the developing brain, as well as in the adult brain. They are self-renewing cells that maintain the capacity to differentiate into all major brain-specific cell types, such as glial cells and neurons. However, it is still unclear whether these cells are capable of gaining full functionality, which is one of the major prerequisites for NSC-based cell replacement strategies of neurological diseases. The ability to establish and maintain polarized excitatory synaptic contacts would be one of the basic requirements for intercellular communication and functional integration into existing neuronal networks. In primary cultures of hippocampal neurons, it has already been shown that synaptogenesis is characterized by a well-ordered, time-dependent targeting and recruitment of pre- and postsynaptic proteins. In this study, we investigated the expression and localization of important pre- and postsynaptic proteins, including Bassoon and synaptophysin, as well as proteins of the ProSAP/Shank family, in differentiating rat fetal mesencephalic NSCs. Moreover, we analyzed the ultrastructural features of neuronal cell-cell contacts during synaptogenesis. We show that NSCs express and localize cytoskeletal and scaffolding molecules of the pre- and postsynaptic specializations in a well-defined temporal order, leading to mature synaptic contacts after 14 days of differentiation. The temporal and spatial pattern of synaptic maturation is comparable to synaptogenesis of hippocampal neurons grown in primary culture. Therefore, with respect to the general ability to create mature synaptic contacts, NSCs seem to be well equipped to potentially compensate for lost or injured brain tissue. Disclosure of potential conflicts of interest is found at the end of this article.     

Regulation of the sigmaE stress response by DegS: how the PDZ domain keeps the protease inactive in the resting state and allows integration of different OMP-derived stress signals upon folding stress

The unfolded protein response of Escherichia coli is triggered by the accumulation of unassembled outer membrane proteins (OMPs) in the cellular envelope. The PDZ-protease DegS recognizes these mislocalized OMPs and initiates a proteolytic cascade that ultimately leads to the sigmaE-driven expression of a variety of factors dealing with folding stress in the periplasm and OMP assembly. The general features of how OMPs activate the protease function of DegS have not yet been systematically addressed. Furthermore, it is unknown how the PDZ domain keeps the protease inactive in the resting state, which is of crucial importance for the functioning of the entire sigmaE stress response. Here we show in atomic detail how DegS is able to integrate the information of distinct stress signals that originate from different OMPs containing a -x-Phe C-terminal motif. A dedicated loop of the protease domain, loop L3, serves as a versatile sensor for allosteric ligands. L3 is capable of interacting differently with ligands but reorients in a conserved manner to activate DegS. Our data also indicate that the PDZ domain directly inhibits protease function in the absence of stress signals by wedging loop L3 in a conformation that ultimately disrupts the proteolytic site. Thus, the PDZ domain and loop L3 of DegS define a novel molecular switch allowing strict regulation of the sigmaE stress response system.     

Wound contamination in cardiac surgery. A systematic quantitative and qualitative study of the bacterial growth in sternal wounds in cardiac surgery patients

OBJECTIVES: To investigate the degree of bacterial contamination in the sternal wound during cardiac surgery and the sternal skin flora after operation in order to increase our understanding of the pathogenesis of sternal wound infections. DESIGN: Prospective study where cultures were taken peri- and postoperatively from sternal wounds and skin. SETTING: University Hospital. PATIENTS: 201 cardiac surgery patients. RESULTS: 89% of the patients grew bacteria from the subcutaneous sternal tissue. 98% of the patients showed bacterial growth on the surrounding skin at the end of the operation. We found both commensal and nosocomial bacteria in the sternal wound. These bacteria had different temporal distribution patterns. Coagulase-negative staphylococci (CoNS) and Propionibacterium acnes (PA) were by far the most prevalent bacteria during and after the operation. Furthermore, 41% of patients had more than 10,000 CFU/pad CoNS on the skin. There was no correlation between length of operation and number of bacteria. Men displayed higher bacterial counts than women on the skin. CONCLUSION: Skin preparation with ethanol/chlorhexidine is unable to suppress the physiological skin flora for the duration of a heart operation. A decrease of CoNS and PA postoperatively can be caused by competitive recolonisation of commensal and nosocomial bacteria.