Transcriptomic and Proteomic Analysis of Arion vulgaris—Proteins for Probably Successful Survival Strategies?

The Spanish slug, Arion vulgaris, is considered one of the hundred most invasive species in Central Europe. The immense and very successful adaptation and spreading of A. vulgaris suggest that it developed highly effective mechanisms to deal with infections and natural predators. Current transcriptomic and proteomic studies on gastropods have been restricted mainly to marine and freshwater gastropods. No transcriptomic or proteomic study on A. vulgaris has been carried out so far, and in the current study, the first transcriptomic database from adult specimen of A. vulgaris is reported. To facilitate and enable proteomics in this non-model organism, a mRNA-derived protein database was constructed for protein identification. A gel-based proteomic approach was used to obtain the first generation of a comprehensive slug mantle proteome. A total of 2128 proteins were unambiguously identified; 48 proteins represent novel proteins with no significant homology in NCBI non-redundant database. Combined transcriptomic and proteomic analysis revealed an extensive repertoire of novel proteins with a role in innate immunity including many associated pattern recognition, effector proteins and cytokine-like proteins. The number and diversity in gene families encoding lectins point to a complex defense system, probably as a result of adaptation to a pathogen-rich environment. These results are providing a fundamental and important resource for subsequent studies on molluscs as well as for putative antimicrobial compounds for drug discovery and biomedical applications

“No one to consult! That is the hardest part” choice-making experiences for prenatal screening tests among Japanese women and their spouses in Austria - A qualitative interview study

Objective: Japan is the only country in the world that allows abortions due to economic reasons but illegalise them due to foetal anomaly. The objective of this study was to explore the choice-making experiences for prenatal screening among Japanese women and their spouses in Austria. Methods: We conducted a qualitative study using semi-structured face-to-face interviews with Japanese women and their spouses in Austria. Data were analysed using thematic analysis. Results: Twenty-five participants (14 women and 11 men) took part in the interviews. Four themes were identified: 1) Knowledge, information and memory; 2) Communication and interactions with health professionals; 3) Reasons for choice; and 4) Emotional support. Participants had limited knowledge and experienced directive counselling. Women expressed negative emotions in the choice-making processes, did not perceive husbands as a source of support and lacked a person to consult. Conclusion: There are common characteristics among East Asian population despite different context and differences found between our Japanese participants and women in other European countries. Practice implication: Proactive interventions aimed at increasing knowledge that help women to develop their preferences and reflect on their values could be further promoted among women of all socio-cultural backgrounds in Austria

Folding correction of ABC-transporter ABCB1 by pharmacological chaperones: a mechanistic concept

Point mutations of ATP‐binding cassette (ABC) proteins are a common cause of human diseases. Available crystal structures indicate a similarity in the architecture of several members of this protein family. Their molecular architecture makes these proteins vulnerable to mutation, when critical structural elements are affected. The latter preferentially involve the two transmembrane domain (TMD)/nucleotide‐binding domain (NBD) interfaces (transmission interfaces), formation of which requires engagement of coupling helices of intracellular loops with NBDs. Both, formation of the active sites and engagement of the coupling helices, are contingent on correct positioning of ICLs 2 and 4 and thus an important prerequisite for proper folding. Here, we show that active site compounds are capable of rescuing P‐glycoprotein (P‐gp) mutants ∆Y490 and ∆Y1133 in a concentration‐dependent manner. These trafficking deficient mutations are located at the transmission interface in pseudosymmetric position to each other. In addition, the ability of propafenone analogs to correct folding correlates with their ability to inhibit transport of model substrates. This finding indicates that folding correction and transport inhibition by propafenone analogs are brought about by binding to the active sites. Furthermore, this study demonstrates an asymmetry in folding correction with cis‐flupentixol, which reflects the asymmetric binding properties of this modulator to P‐gp. Our results suggest a mechanistic model for corrector action in a model ABC transporter based on insights into the molecular architecture of these transporters.© 2017 The Author

Synthesis of ortho-Functionalized 4-Aminomethylpyridazines as Substrate-Like Semicarbazide-Sensitive Amine Oxidase Inhibitors

A series of 4-aminomethylpyridazines and -pyridazin-3(2H)-ones (“diaza-benzylamines”), bearing alkylamino side chains in ortho position relative to the CH2NH2 unit, was synthesized by catalytic hydrogenation of the corresponding nitriles in strongly acidic medium. N-Benzyl protecting groups either at the pyridazinone ring nitrogen or at an exocyclic nitrogen were selectively removed hydrogenolytically or by treatment with a Lewis acid. The new compounds were tested in vitro for semicarbazide-sensitive amine oxidase (SSAO) inhibitory activity and 4-(aminomethyl)-N,N-diethylpyridazine-3,5-diamine (22) was found to be the most active representative

Plasticity of fibroblasts demonstrated by tissue-specific and function-related proteome profiling - Additional files

Additional files to: Slany, Astrid, Anastasia Meshcheryakova, Agnes Beer, Hendrik Ankersmit, Verena Paulitschke, and Christopher Gerner. 2014. "Plasticity of Fibroblasts Demonstrated by Tissue-Specific and Function-Related Proteome Profiling." Clinical Proteomics 11 (1): 41. doi:10.1186/1559-0275-11-41. Additional file 1: Figure S1: FACS analysis of primary lung fibroblasts obtained from non-cancerous and cancerous tissue areas. Cells were characterized by FACS analysis, which showed that cells were positive for fibroblast-specific markers CD90, but negative for leukocyte, endothelial cell and hematopoietic stem cell markers CD45, CD31 and CD34 respectively. All samples contained cells which were inflammatory activated, as demonstrated by positive CD54-staining. A certain amount of the cells showed also a positive staining for α-SMA, characterizing a myofibroblast phenotype of cancer-associated fibroblasts. Additional file 2: Table S1: Proteome profile of NHLF. Accession, Swiss-Prot accession numbers; name, protein names; peptides, the number of distinct peptides identified for each protein. Additional file 3: Table S2: Proteome profile of primary lung fibroblasts. Accession, Swiss-Prot accession numbers; name, protein names; peptides, the number of distinct peptides identified for each protein. Additional file 4: Table S3: Proteome profile of skin fibroblasts. Accession, Swiss-Prot accession numbers; name, protein names; peptides, the number of distinct peptides identified for each protein. Additional file 5: Table S4: Proteome profile of BM fibroblasts. Accession, Swiss-Prot accession numbers; name, protein names; peptides, the number of distinct peptides identified for each protein. Additional file 6: Table S5: Proteome profile of NHLF treated with IL-1β. Accession, Swiss-Prot accession numbers; name, protein names; peptides, the number of distinct peptides identified for each protein. Additional file 7: Table S6: Proteome profile of skin fibroblasts treated with IL-1β. Accession, Swiss-Prot accession numbers; name, protein names; peptides, the number of distinct peptides identified for each protein. Additional file 8: Table S7: Proteome profile of BM fibroblasts treated with IL-1β. Accession, Swiss-Prot accession numbers; name, protein names; peptides, the number of distinct peptides identified for each protein. Additional file 9: Table S9: Proteome profile of melanoma-associated fibroblasts. Accession, Swiss-Prot accession numbers; name, protein names; peptides, the number of distinct peptides identified for each protein. Additional file 10: Table S10: Proteome profile of multiple myeloma-associated fibroblasts. Accession, Swiss-Prot accession numbers; name, protein names; peptides, the number of distinct peptides identified for each protein. Additional file 11: Table S11: Proteome profile of HCC-associated fibroblasts. Accession, Swiss-Prot accession numbers; name, protein names; peptides, the number of distinct peptides identified for each protein. Additional file 12: Table S8: Proteome profile of lung carcinoma-associated fibroblasts. Accession, Swiss-Prot accession numbers; name, protein names; peptides, the number of distinct peptides identified for each protein. Additional file 13: Table S12: emPAI values for all proteins listed in Figure 2. For each protein the emPAI values determined by us in the different sub-cellular fractions (sn, cell supernatant; cyt, cytoplasmic fraction; nuc, nuclear fraction) of the respective cell type and cell state are indicated. AccNr, Swiss-Prot accession number. Additional file 14: Table S13: emPAI values for all proteins listed in Table 1. For each protein the emPAI values determined by us in the different sub-cellular fractions (sn, cell supernatant; cyt, cytoplasmic fraction; nuc, nuclear fraction) of the respective cell type and cell state are indicated. AccNr, Swiss-Prot accession number

The Human Factor: Behavioral and Neural Correlates of Humanized Perception in Moral Decision Making

The extent to which people regard others as full-blown individuals with mental states (“humanization”) seems crucial for their prosocial motivation towards them. Previous research has shown that decisions about moral dilemmas in which one person can be sacrificed to save multiple others do not consistently follow utilitarian principles. We hypothesized that this behavior can be explained by the potential victim’s perceived humanness and an ensuing increase in vicarious emotions and emotional conflict during decision making. Using fMRI, we assessed neural activity underlying moral decisions that affected fictitious persons that had or had not been experimentally humanized. In implicit priming trials, participants either engaged in mentalizing about these persons (Humanized condition) or not (Neutral condition). In subsequent moral dilemmas, participants had to decide about sacrificing these persons’ lives in order to save the lives of numerous others. Humanized persons were sacrificed less often, and the activation pattern during decisions about them indicated increased negative affect, emotional conflict, vicarious emotions, and behavioral control (pgACC/mOFC, anterior insula/IFG, aMCC and precuneus/PCC). Besides, we found enhanced effective connectivity between aMCC and anterior insula, which suggests increased emotion regulation during decisions affecting humanized victims. These findings highlight the importance of others’ perceived humanness for prosocial behavior - with aversive affect and other-related concern when imagining harming more “human-like” persons acting against purely utilitarian decisions.© Majdandžić et a

Combined Metabolomic Analysis of Plasma and Urine Reveals AHBA, Tryptophan and Serotonin Metabolism as Potential Risk Factors in Gestational Diabetes Mellitus (GDM)

Gestational diabetes mellitus during pregnancy has severe implications for the health of the mother and the fetus. Therefore, early prediction and an understanding of the physiology are an important part of prenatal care. Metabolite profiling is a long established method for the analysis and prediction of metabolic diseases. Here, we applied untargeted and targeted metabolomic protocols to analyze plasma and urine samples of pregnant women with and without GDM. Univariate and multivariate statistical analyses of metabolomic profiles revealed markers such as 2-hydroxybutanoic acid (AHBA), 3-hydroxybutanoic acid (BHBA), amino acids valine and alanine, the glucose-alanine-cycle, but also plant-derived compounds like sitosterin as different between control and GDM patients. PLS-DA and VIP analysis revealed tryptophan as a strong variable separating control and GDM. As tryptophan is biotransformed to serotonin we hypothesized whether serotonin metabolism might also be altered in GDM. To test this hypothesis we applied a method for the analysis of serotonin, metabolic intermediates and dopamine in urine by stable isotope dilution direct infusion electrospray ionization mass spectrometry (SID-MS). Indeed, serotonin and related metabolites differ significantly between control and GDM patients confirming the involvement of serotonin metabolism in GDM. Clustered correlation coefficient visualization of metabolite correlation networks revealed the different metabolic signatures between control and GDM patients. Eventually, the combination of selected blood plasma and urine sample metabolites improved the AUC prediction accuracy to 0.99. The detected GDM candidate biomarkers and the related systemic metabolic signatures are discussed in their pathophysiological context. Further studies with larger cohorts are necessary to underpin these observations.© 2017 Leitner, Fragner, Danner, Holeschofsky, Leitner, Tischler, Doerfler, Bachmann, Sun, Jaeger, Kautzky-Willer and Weckwert

Plasticity of fibroblasts demonstrated by tissue-specific and function-related proteome profiling

Background: Fibroblasts are mesenchymal stromal cells which occur in all tissue types. While their main function is related to ECM production and physical support, they are also important players in wound healing, and have further been recognized to be able to modulate inflammatory processes and support tumor growth. Fibroblasts can display distinct phenotypes, depending on their tissue origin, as well as on their functional state. Results: In order to contribute to the proteomic characterization of fibroblasts, we have isolated primary human fibroblasts from human skin, lung and bone marrow and generated proteome profiles of these cells by LC-MS/MS. Comparative proteome profiling revealed characteristic differences therein, which seemed to be related to the cell’s tissue origin. Furthermore, the cells were treated in vitro with the pro-inflammatory cytokine IL-1beta. While all fibroblasts induced the secretion of Interleukins IL-6 and IL-8 and the chemokine GRO-alpha, other inflammation-related proteins were up-regulated in an apparently tissue-dependent manner. Investigating fibroblasts from tumorous tissues of skin, lung and bone marrow with respect to such inflammation-related proteins revealed hardly any conformity but rather individual and tumor type-related variations. However, apparent up-regulation of IGF-II, PAI-1 and PLOD2 was observed in melanoma-, lung adenocarcinoma- and multiple myeloma-associated fibroblasts, as well as in hepatocellular carcinoma-associated fibroblasts. Conclusions: Inflammation-related proteome alterations of primary human fibroblasts were determined by the analysis of IL-1beta treated cells. Tumor-associated fibroblasts from different tissue types hardly showed signs of acute inflammation but displayed characteristic functional aberrations potentially related to chronic inflammation. The present data suggest that the state of the tumor microenvironment is relevant for tumor progression and targeted treatment of tumor-associated fibroblasts may support anti-cancer strategies

On Orthogonal Projections for Dimension Reduction and Applications in Augmented Target Loss Functions for Learning Problems

The use of orthogonal projections on high-dimensional input and target data in learning frameworks is studied. First, we investigate the relations between two standard objectives in dimension reduction, preservation of variance and of pairwise relative distances. Investigations of their asymptotic correlation as well as numerical experiments show that a projection does usually not satisfy both objectives at once. In a standard classification problem, we determine projections on the input data that balance the objectives and compare subsequent results. Next, we extend our application of orthogonal projections to deep learning tasks and introduce a general framework of augmented target loss functions. These loss functions integrate additional information via transformations and projections of the target data. In two supervised learning problems, clinical image segmentation and music information classification, the application of our proposed augmented target loss functions increases the accuracy.© The Author(s) 201

R-Modafinil exerts weak effects on spatial memory acquisition and dentate gyrus synaptic plasticity

Modafinil is a wake promoting drug approved for clinical use and also has cognitive enhancing properties. Its enantiomer R-Modafinil (R-MO) is not well studied in regard to cognitive enhancing properties. Hence we studied its effect in a spatial memory paradigm and its possible effects on dentate gyrus long-term potentiation (DG-LTP). Clinically relevant doses of R-MO, vehicle dimethyl sulfoxide (DMSO) or saline were administered for three days during the hole-board test and in in vivo DG-LTP. Synaptic levels of dopamine receptors D1R, D2R, dopamine transporter (DAT), and its phosphorylated form (ph-DAT) in DG tissue 4 h after LTP induction were quantified by western blot analysis. Monoamine reuptake and release assays were performed by using transfected HEK-293 cells. Possible neurotoxic side effects on general behaviour were also studied. R-MO at both doses significantly enhanced spatial reference memory during the last training session and during memory retrieval compared to DMSO vehicle but not when compared to saline treated rats. Similarly, R-MO rescues DG-LTP from impairing effects of DMSO. DMSO reduced memory performance and LTP magnitude when compared to saline treated groups. The synaptic DR1 levels in R-MO groups were significantly decreased compared to DMSO group but were comparable with saline treated animals. We found no effect of R-MO in neurotoxicity tests. Thus, our results support the notion that LTP-like synaptic plasticity processes could be one of the factors contributing to the cognitive enhancing effects of spatial memory traces. D1R may play an important regulatory role in these processes.© 2017 Shanmugasundaram et a