Reg-1α, a New Substrate of Calpain-2 Depending on Its Glycosylation Status

Reg-1α/lithostathine, a protein mainly associated with the digestive system, was previously shown to be overexpressed in the pre-clinical stages of Alzheimer’s disease. In vitro, the glycosylated protein was reported to form fibrils at physiological pH following the proteolytic action of trypsin. However, the nature of the protease able to act in the central nervous system is unknown. In the present study, we showed that Reg-1α can be cleaved in vitro by calpain-2, the calcium activated neutral protease, overexpressed in neurodegenerative diseases. Using chemical crosslinking experiments, we found that the two proteins can interact with each other. Identification of the cleavage site using mass spectrometry, between Gln⁴ and Thr⁵, was found in agreement with the in silico prediction of the calpain cleavage site, in a position different from the one reported for trypsin, i.e., Arg¹¹-Ile¹² peptide bond. We showed that the cleavage was impeded by the presence of the neighboring glycosylation of Thr⁵. Moreover, in vitro studies using electron microscopy showed that calpain-cleaved protein does not form fibrils as observed after trypsin cleavage. Collectively, our results show that calpain-2 cleaves Reg-1α in vitro, and that this action is not associated with fibril formation.     

Proteomic Biomarkers of the Apnea Hypopnea Index and Obstructive Sleep Apnea: Insights into the Pathophysiology of Presence,Severity, and Treatment Response

Obstructive sleep apnea (OSA), a disease associated with excessive sleepiness and increased cardiovascular risk, affects an estimated 1 billion people worldwide. The present study examined proteomic biomarkers indicative of presence, severity, and treatment response in OSA. Participants (n = 1391) of the Stanford Technology Analytics and Genomics in Sleep study had blood collected and completed an overnight polysomnography for scoring the apnea–hypopnea index (AHI). A highly multiplexed aptamer-based array (SomaScan) was used to quantify 5000 proteins in all plasma samples. Two separate intervention-based cohorts with sleep apnea (n = 41) provided samples pre- and post-continuous/positive airway pressure (CPAP/PAP). Multivariate analyses identified 84 proteins (47 positively, 37 negatively) associated with AHI after correction for multiple testing. Of the top 15 features from a machine learning classifier for AHI ≥ 15 vs. AHI < 15 (Area Under the Curve (AUC) = 0.74), 8 were significant markers of both AHI and OSA from multivariate analyses. Exploration of pre- and post-intervention analysis identified 5 of the 84 proteins to be significantly decreased following CPAP/PAP treatment, with pathways involving endothelial function, blood coagulation, and inflammatory response. The present study identified PAI-1, tPA, and sE-Selectin as key biomarkers and suggests that endothelial dysfunction and increased coagulopathy are important consequences of OSA, which may explain the association with cardiovascular disease and stroke.     

Tumor Infiltration with CD20+CD73+ B Cells Correlates with Better Outcome in Colorectal Cancer

Immunotherapy has become increasingly important in the treatment of colorectal cancer (CRC). Currently, CD73, also known as ecto-5′-nucleotidase (NT5E), has gained considerable interest as a potential therapeutic target. CD73 is one of the key enzymes catalyzing the conversion of extracellular ATP into adenosine, which in turn exerts potent immune suppressive effects. However, the role of CD73 expression on various cell types within the CRC tumor microenvironment remains unresolved. The expression of CD73 on various cell types has been described recently, but the role of CD73 on B-cells in CRC remains unclear. Therefore, we analyzed CD73 on B-cells, especially on tumor-infiltrating B-cells, in paired tumor and adjacent normal tissue samples from 62 eligible CRC patients. The highest expression of CD73 on tumor-infiltrating B-cells was identified on class-switched memory B-cells, followed by naive B-cells, whereas no CD73 expression was observed on plasmablasts. Clinicopathological correlation analysis revealed that higher CD73⁺ B-cells infiltration in the CRC tumors was associated with better overall survival. Moreover, metastasized patients showed a significantly decreased number of tumor-infiltrating CD73⁺ B-cells. Finally, neoadjuvant therapy correlated with reduced CD73⁺ B-cell numbers and CD73 expression on B-cells in the CRC tumors. As promising new immune therapies are being developed, the role of CD73⁺ B-cells and their subsets in the development of colorectal cancer should be further explored to find new therapeutic options.     

Characterization of a Cutibacterium acnes Camp Factor 1-Related Peptide as a New TLR-2 Modulator in In Vitro and Ex Vivo Models of Inflammation

Cutibacterium acnes (C. acnes) has been implicated in inflammatory acne where highly mutated Christie–Atkins–Munch–Petersen factor (CAMP)1 displays strong toll like receptor (TLR)-2 binding activity. Using specific antibodies, we showed that CAMP1 production was independent of C. acnes phylotype and involved in the induction of inflammation. We confirmed that TLR-2 bound both mutated and non-mutated recombinant CAMP1, and peptide array analysis showed that seven peptides (A14, A15, B1, B2, B3, C1 and C3) were involved in TLR-2 binding, located on the same side of the three-dimensional structure of CAMP1. Both mutated and non-mutated recombinant CAMP1 proteins induced the production of C-X-C motif chemokine ligand interleukin (CXCL)8/(IL)-8 in vitro in keratinocytes and that of granulocyte macrophage-colony stimulating factor (GM-CSF), tumor necrosis factor (TNF)-α, IL-1β and IL-10 in ex vivo human skin explants. Only A14, B1 and B2 inhibited the production of CXCL8/IL-8 by keratinocytes and that of (GM-CSF), TNF-α, IL-1β and IL-10 in human skin explants stimulated with rCAMP1 and C. acnes. Following pretreatment with B2, RNA sequencing on skin explants identified the 10 genes displaying the strongest differential expression as IL6, TNF, CXCL1, CXCL2, CXCL3, CXCL8, IL-1β, chemokine ligand (CCL)2, CCL4 and colony stimulating factor (CSF)2. We, thus, identified a new CAMP1-derived peptide as a TLR-2 modulator likely to be a good candidate for clinical evaluation.     

家具与衣服

Couch Sleeper、Home Traveller、Lingerie Locker是Anne Lorenz在Karlsruhe艺术设计学院时毕业作品的一部分。可能因为女性天生对衣服就有很高的敏锐度,Anne Lorenz一直着迷于研究家具与衣物之间的关系,并不断探讨它们之间的形式与功能。     

Effects of exercising before versus after eating on dieting and exercise evaluations: A preliminary investigation.

Psychological processes may play a role in the evaluation of the effectiveness of exercise and subsequent food intake. In order to further investigate this phenomenon, the effects of the timing of exercise relative to an eating opportunity were evaluated. Female undergraduate participants who were of average weight and did not exercise regularly were randomly assigned to one of three conditions (exercise before eating [n = 10], exercise after eating [n = 11], or no exercise [n = 12]). Expectations of the effectiveness of the exercise, value of dieting, and intake were assessed. Participants who exercised after eating had higher expectations of the effectiveness of the exercise than those who exercised before eating, while those who exercised before eating reported valuing dieting more than controls. No effects on intake emerged; therefore, there appears to be a disconnect between dieting appraisals and actual eating behaviour. The results are discussed in relation to theories on conflicting goals. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Cell design in bacteria as a convex optimization problem

In this paper, we investigate the prediction of the cell composition of bacteria with respect to their medium. By modeling the bacterium as an interconnection of subsystems, the problem is written as a non-smooth convex optimization problem equivalent to a Linear Programming feasibility problem. We then obtain a new method, called Resource Balance Analysis (RBA), predicting the distribution of the available resources in the medium among the various cellular subsystems. Beyond its predictive capability, the proposed approach grasps some fundamental aspects of the bacterium physiology by including a refined model. This method reveals the existence of an intrinsic bottleneck in the system resource distribution of the bacterium, leading to the existence of a structural limitation of its growth rate which can be predicted. RBA is also able to predict the configuration of the metabolic network for a given medium at steady-state regimen which nicely fits the available experimental results for the gram-positive model bacterium Bacillus subtilis.     

Finite element model updating and structural damage identification using OMAX data

The main limitations in the finite element (FE) model updating technique lie in the ability of the FE model to represent the true behavior of the structure (modelling problem), and in the ability to identify enough modal parameters with sufficient accuracy, especially for large structures that are tested in operational conditions (identification problem). In this paper, the identification problem is solved with an OMAX approach, where an artificial force is used in operational conditions and a structural model is identified that takes both the forced and the ambient excitation into account. From an extensive case study on a real three-span bridge, it is observed that, while updating the FE model using the experimental output-only data yields a good fit, discrepancies show up when the more extensive set of OMAX data is used for validation, or even for updating. It can be concluded that an OMAX approach not only increases the well-posedness of the updating problem, it also allows to detect potential inaccuracies in the FE model.