Pancreatic islet cell stress induced by insulin-degrading enzyme deficiency promotes islet regeneration and protection from autoimmune diabetes.

Appropriate tuning of protein homeostasis through mobilization of the unfolded protein response (UPR) is key to the capacity of pancreatic beta cells to cope with highly variable demand for insulin synthesis. An efficient UPR ensures a sufficient beta cell mass and secretory output but can also affect beta cell resilience to autoimmune aggression. The factors regulating protein homeostasis in the face of metabolic and immune challenges are insufficiently understood. We examined beta cell adaptation to stress in mice deficient for insulin-degrading enzyme (IDE), a ubiquitous protease with high affinity for insulin and genetic association with type 2 diabetes. IDE deficiency induced a low-level UPR in both C57BL/6 and autoimmune non-obese diabetic (NOD) mice, associated with rapamycin-sensitive beta cell proliferation strongly enhanced by proteotoxic stress. Moreover, in NOD mice, IDE deficiency protected from spontaneous diabetes and triggered an additional independent pathway, conditional on the presence of islet inflammation but inhibited by proteotoxic stress, highlighted by strong upregulation of regenerating islet-derived protein 2, a protein attenuating autoimmune inflammation. Our findings establish a key role of IDE in islet cell protein homeostasis, identify a link between low-level UPR and proliferation, and reveal an UPR-independent anti-inflammatory islet cell response uncovered in the absence of IDE of potential interest in autoimmune diabetes.

The Digital Brain Tumour Atlas, an open histopathology resource.

Currently, approximately 150 different brain tumour types are defined by the WHO. Recent endeavours to exploit machine learning and deep learning methods for supporting more precise diagnostics based on the histological tumour appearance have been hampered by the relative paucity of accessible digital histopathological datasets. While freely available datasets are relatively common in many medical specialties such as radiology and genomic medicine, there is still an unmet need regarding histopathological data. Thus, we digitized a significant portion of a large dedicated brain tumour bank based at the Division of Neuropathology and Neurochemistry of the Medical University of Vienna, covering brain tumour cases from 1995-2019. A total of 3,115 slides of 126 brain tumour types (including 47 control tissue slides) have been scanned. Additionally, complementary clinical annotations have been collected for each case. In the present manuscript, we thoroughly discuss this unique dataset and make it publicly available for potential use cases in machine learning and digital image analysis, teaching and as a reference for external validation.

Isolation and Cultivation of Adipose-Derived Mesenchymal Stem Cells Originating from the Infrapatellar Fat Pad Differentiated with Blood Products: Method and Protocol.

Adipose-derived mesenchymal stem cells (ASCs) are a promising source for clinical application in regenerative orthopedics. ASCs derived from the infra-patellar fat pad (IFP)-a distinct adipose structure in the knee-show superior regenerative potential compared to subcutaneous-fat-derived cells. Furthermore, it has been shown that blood products enhance ASCs' viability. A major challenge for clinical translation of both ASCs and blood products is the low comparability of obtained data due to non-standardized harvesting, isolation and preparation methods. The aim of this method-paper is to provide reproducible protocols to help standardize basic research in the field to build a sound basis for clinical translation with an emphasize on practicability. The presented protocols include (i) ASC isolation from the IFP, (ii) blood product preparation and (iii) ASC incubation with blood products.

Counseling in Vape Shops: A Survey of Vape Shop Managers in Switzerland.

Vaporizers (e-cigarettes) are the most common smoking cessation aids in Switzerland, but we do not know what information customers receive in vape shops. We surveyed vape-shop managers to find out what recommendations they make to their customers. An interdisciplinary group developed the questionnaire. Respondents self-reported their smoking history, demographics, and the recommendations they thought they would give to hypothetical customers in clinical vignettes. We also queried if they collaborated with health care professionals. Of those contacted, 53.8% (70/130) of vape-shop managers responded, and 52.3% (68/130) were included in the final analysis. Managers were mostly male and ex-smokers who switched to vaporizers; 60.3% would encourage a hypothetical smoker with high nicotine dependence to start with the highest possible nicotine concentration when switching to vaporizers. For this smoker, 36.9% would recommend high (≥15 mg/mL), 32.3% medium (6-14 mg/mL), and 3.1% low (1-5 mg/mL) nicotine concentrations. The rest adapted their recommendations to fit the customer or device; 76.5% reported that physicians referred customers to them, and 78.8% would attend a course given by experts in the field of vaporizers and smoking cessation. Vape-shop managers varied widely in the recommendations they gave customers. Most reported ongoing collaboration with health care professionals and were interested in improving their counselling skills through training.

[Helpful Factors of Ambulant Art Therapy in the Group and Changes of Experiences in Psychosomatic Patients]. / Wirkfaktoren ambulanter Kunsttherapie in der Gruppe und Veränderungen im Erleben psychosomatischer Patienten.

The aim of this study was to analyze the experiences of patients suffering from mostly chronic psychosomatic disorders in an ambulant art therapy in the group. Especially, the focus was on the experienced changes, helpful factors and specifics of the therapy as well as on the experienced benefit. For this, 30 patients were interviewed in a semi-standardized way. Additionally, the symptom-based strain was psychometrically recorded in a part of the patients (21) at the beginning of the therapy and after at least 6 months of participation. The evaluation of those interviews with the qualitative analysis of the therapy subjects surrendered an improvement of the health state in most of the participants. Especially group factors, art as a mean of communication, becoming aware of feelings but also diversion and fun were proved to be beneficial. The art therapy also serves for structuring the week as well as a contact point and a resource in the interpersonal communication of everyday life. Nearly all of the patients referred to some important turning point pictures. Mostly, the benefit was valued as being high. But, in contrast, the psychometric measure did not show any significant change. The results emphasize the stabilizing function of art therapy in the examined patients, whereat the classification of the psychometric result is complicated by the absence of a control group.

Endoplasmic reticulum targeting alters regulation of expression and antigen presentation of proinsulin.

Peptide ligands presented by MHC class I (MHC-I) molecules are produced by degradation of cytosolic and nuclear, but also endoplasmic reticulum (ER)-resident, proteins by the proteasome. However, Ag processing of ER proteins remains little characterized. Studying processing and presentation of proinsulin, which plays a pivotal role in autoimmune diabetes, we found that targeting to the ER has profound effects not only on how proinsulin is degraded, but also on regulation of its cellular levels. While proteasome inhibition inhibited degradation and presentation of cytosolic proinsulin, as expected, it reduced the abundance of ER-targeted proinsulin. This targeting and protein modifications modifying protein half-life also had profound effects on MHC-I presentation and proteolytic processing of proinsulin. Thus, presentation of stable luminal forms was inefficient but enhanced by proteasome inhibition, whereas that of unstable luminal forms and of a cytosolic form were more efficient and compromised by proteasome inhibitors. Distinct stability of peptide MHC complexes produced from cytosolic and luminal proinsulin suggests that different proteolytic activities process the two Ag forms. Thus, both structural features and subcellular targeting of Ags can have strong effects on the processing pathways engaged by MHC-I-restricted Ags, and on the efficiency and regulation of their presentation.

No major role for insulin-degrading enzyme in antigen presentation by MHC molecules.

Antigen presentation by MHC class I molecules requires degradation of epitope source proteins in the cytosol. Although the preeminent role of the proteasome is clearly established, evidence suggesting a significant role for proteasome-independent generation of class I ligands has been reported repeatedly. However, an enzyme responsible for such a role has not been identified. Recently insulin-degrading enzyme (IDE) was shown to produce an antigenic peptide derived from the tumor antigen MAGE-A3 in an entirely proteasome-independent manner, raising the question of the global impact of IDE in MHC class I antigen processing. Here we report that IDE knockdown in human cell lines, or knockout in two different mouse strains, has no effect on cell surface expression of various MHC class I molecules, including allomorphs such as HLA-A3 and HLA-B27 suggested to be loaded in an at least a partly proteasome-independent manner. Moreover, reduced or absent IDE expression does not affect presentation of five epitopes including epitopes derived from beta amyloid and proinsulin, two preferred IDE substrates. Thus, IDE does not play a major role in MHC class I antigen processing, confirming the dominant and almost exclusive role of the proteasome in cytosolic production of MHC class I ligands.

Social information and personal interests modulate neural activity during economic decision-making.

In the present study we employed electrophysiological recordings to investigate the levels of processing at which positive and negative descriptions of other people bias social decision-making in a game in which participants accepted or rejected economic offers. Besides social information, we manipulated the fairness of the assets distribution, whether offers were advantageous or not for the participant and the uncertainty of the game context. Results show that a negative description of the interaction partner enhanced the medial frontal negativity (MFN) in an additive manner with fairness evaluations. The description of the partner interacted with personal benefit considerations, showing that this positive or negative information only biased the evaluation of offers when they did not favor the participant. P300 amplitudes were enhanced by advantageous offers, suggesting their heightened motivational significance at later stages of processing. Throughout all stages, neural activity was enhanced with certainty about the personal assignments of the split. These results provide new evidence on the importance of interpersonal information and considerations of self-interests relative to others in decision-making situations.

Social information and economic decision-making in the ultimatum game.

The present study tested how social information about the proposer biases responders' choices of accepting or rejecting real monetary offers in a classic ultimatum game (UG) and whether this impact is heightened by the uncertainty of the context. Participants in our study conducted a one-shot UG in which their responses had direct consequences on how much money they earned. We used trait-valenced words to provide information about the proposers' personal characteristics. The results show higher acceptance rates for offers preceded by positive words than for those preceded by negative words. In addition, the impact of this information was higher in the uncertain than in the certain context. This suggests that when deciding whether or not to take money from someone, people take into account what they know about the person they are interacting with. Such non-rational bias is stronger in an uncertain context.

Polo-like kinase 1 is a potential therapeutic target in human melanoma.

Exploration of the human melanoma cell-cycle pathway can lead to identification of new therapeutic targets. By gene set enrichment analysis, we identified the cell-cycle pathway and its member polo-like kinase 1 (Plk-1) to be significantly overexpressed in primary melanomas and in melanoma metastases. In vitro expression of Plk-1 was peaked at the G2/M phase of the cell cycle. Plk-1 knockdown/inhibition led to induction of apoptosis, which was caspase-3/8-dependent and p53-independent, and involved BID and Bcl-2 proteins. Comparative genomic hybridization/single-nucleotide polymorphism arrays showed no genetic alteration in the Plk-1 locus. Previous suggestions and significant enrichment of the mitogen-activated protein kinase (MAPK) signaling pathway pointed to potential regulation of Plk-1 by MAPK signaling. Inhibition of this pathway resulted in decreased Plk-1 expression as a consequence of G1 cell-cycle arrest rather than direct regulation of Plk-1. Inhibition of MAPK and Plk-1 had an additive effect on reduced cell viability. This study shows that in human melanoma, Plk-1 expression is dynamically regulated during the cell cycle, knockdown of Plk-1 leads to apoptotic cell death, and that a combination of Plk-1 and MAPK inhibition has an additive effect on melanoma cell viability. We conclude that combined inhibition of Plk-1 and MAPK could be a potentially attractive strategy in melanoma therapy.

Social expectations bias decision-making in uncertain inter-personal situations.

Understanding the role that social cues have on interpersonal choice, and their susceptibility to contextual effects, is of core importance to models of social decision-making. Language, on the other hand, is one of the main means of communication during social interactions in our culture. The present experiments tested whether positive and negative linguistic descriptions of alleged partners in a modified Ultimatum Game biased decisions made to the same set of offers, and whether the contextual uncertainty of the game modulated this biasing effect. The results showed that in an uncertain context, the same offers were accepted with higher probability when they were preceded by positive rather than by negative valenced trait-words. Participants also accepted fair offers with higher probability than unfair offers, but this effect did not interact with the valence of the social descriptive words. In addition, the speed of the decision was affected by valence: acceptance choices were faster when they followed a positive adjective, whereas rejection responses were faster after a negative-valenced word. However, these effects were highly reduced when the uncertainty was eliminated from the game. This suggests that positive and negative relevant social information can bias decisions made to the same pieces of evidence during interpersonal interactions, but that this mainly takes place when the uncertainty associated with the choices is high.

Beta cell antigens in type 1 diabetes: triggers in pathogenesis and therapeutic targets.

Recognition of pancreatic beta cell antigens by autoreactive T lymphocytes plays a central role in the pathogenesis of insulin-dependent type 1 diabetes. Recent results suggest that non-conventional antigenic epitope processing and presentation may contribute to triggering and maintaining autoreactive responses. Moreover, promising results raise hope that autoantigens may become safe and specific therapeutics for type 1 diabetes in the future.

Phase II trial of a toll-like receptor 9-activating oligonucleotide in patients with metastatic melanoma.

PURPOSE: The recent identification of toll-like receptors (TLRs) and respective ligands allows the evaluation of novel dendritic cell (DC) -activating strategies. Stimulation of TLR9 directly activates human plasmacytoid DCs (PDCs) and indirectly induces potent innate immune responses in preclinical tumor models. We performed an open-label, multicenter, single-arm, phase II pilot trial with a TLR9-stimulating oligodeoxynucleotide in melanoma patients. PATIENTS AND METHODS: Patients with unresectable stage IIIb/c or stage IV melanoma received 6 mg PF-3512676 weekly by subcutaneous injection for 24 weeks or until disease progression to evaluate safety as well as clinical and immunologic activity. Clinical and laboratory safety assessments were performed weekly; blood samples for immunological measurements were taken every 8 weeks. Tumor measurements were performed according to Response Evaluation Criteria in Solid Tumors. RESULTS: Twenty patients received PF-3512676 for a mean of 10.9 weeks with a mean of 10.7 injections. Laboratory and nonlaboratory adverse events were limited, transient, and did not result in any withdrawals. Two patients experienced a confirmed partial response; one response is ongoing for 140+ weeks. Three patients experienced stable disease. Immunologic measurements revealed induction of an activated phenotype of PDC, elevation of serum levels of 2',5'-oligoadenylate, a surrogate marker of type I interferon production, and significant stimulation of natural killer cell cytotoxicity (the latter was associated with clinical benefit). CONCLUSION: These results indicate that TLR9-targeted therapy can stimulate innate immune responses in cancer patients, identify biomarkers that may be associated with TLR9-induced tumor regression, and encourage the design of follow-up studies to evaluate the ability of this therapeutic approach to target human cancer.