Encoding and transducing the synaptic or extrasynaptic origin of NMDA receptor signals to the nucleus.

The activation of N-methyl-D-aspartate-receptors (NMDARs) in synapses provides plasticity and cell survival signals, whereas NMDARs residing in the neuronal membrane outside synapses trigger neurodegeneration. At present, it is unclear how these opposing signals are transduced to and discriminated by the nucleus. In this study, we demonstrate that Jacob is a protein messenger that encodes the origin of synaptic versus extrasynaptic NMDAR signals and delivers them to the nucleus. Exclusively synaptic, but not extrasynaptic, NMDAR activation induces phosphorylation of Jacob at serine-180 by ERK1/2. Long-distance trafficking of Jacob from synaptic, but not extrasynaptic, sites depends on ERK activity, and association with fragments of the intermediate filament α-internexin hinders dephosphorylation of the Jacob/ERK complex during nuclear transit. In the nucleus, the phosphorylation state of Jacob determines whether it induces cell death or promotes cell survival and enhances synaptic plasticity.

The COP9 signalosome: A versatile regulatory hub of Cullin-RING ligases.

The COP9 signalosome (CSN) is a universal regulator of Cullin-RING ubiquitin ligases (CRLs) - a family of modular enzymes that control various cellular processes via timely degradation of key signaling proteins. The CSN, with its eight-subunit architecture, employs multisite binding of CRLs and inactivates CRLs by removing a small ubiquitin-like modifier named neural precursor cell-expressed, developmentally downregulated 8 (Nedd8). Besides the active site of the catalytic subunit CSN5, two allosteric sites are present in the CSN, one of which recognizes the substrate recognition module and the presence of CRL substrates, and the other of which can 'glue' the CSN-CRL complex by recruitment of inositol hexakisphosphate. In this review, we present recent findings on the versatile regulation of CSN-CRL complexes.

Fault roughness controls injection-induced seismicity.

Surface roughness ubiquitously prevails in natural faults across various length scales. Despite extensive studies highlighting the important role of fault geometry in the dynamics of tectonic earthquakes, whether and how fault roughness affects fluid-induced seismicity remains elusive. Here, we investigate the effects of fault geometry and stress heterogeneity on fluid-induced fault slip and associated seismicity characteristics using laboratory experiments and numerical modeling. We perform fluid injection experiments on quartz-rich sandstone samples containing either a smooth or a rough fault. We find that geometrical roughness slows down injection-induced fault slip and reduces macroscopic slip velocities and fault slip-weakening rates. Stress heterogeneity and roughness control hypocenter distribution, frequency-magnitude characteristics, and source mechanisms of injection-induced acoustic emissions (AEs) (analogous to natural seismicity). In contrast to smooth faults where injection-induced AEs are uniformly distributed, slip on rough faults produces spatially localized AEs with pronounced non-double-couple source mechanisms. We demonstrate that these clustered AEs occur around highly stressed asperities where induced local slip rates are higher, accompanied by lower Gutenberg-Richter b-values. Our findings suggest that real-time monitoring of induced microseismicity during fluid injection may allow identifying progressive localization of seismic activity and improve forecasting of runaway events.

Gastric Epithelial Barrier Disruption, Inflammation and Oncogenic Signal Transduction by Helicobacter pylori.

Helicobacter pylori exemplifies one of the most favourable bacterial pathogens worldwide. The bacterium colonizes the gastric mucosa in about half of the human population and constitutes a major risk factor for triggering gastric diseases such as stomach cancer. H. pylori infection represents a prime example of chronic inflammation and cancer-inducing bacterial pathogens. The microbe utilizes a remarkable set of virulence factors and strategies to control cellular checkpoints of inflammation and oncogenic signal transduction. This chapter emphasizes on the pathogenicity determinants of H. pylori such as the cytotoxin-associated genes pathogenicity island (cagPAI)-encoded type-IV secretion system (T4SS), effector protein CagA, lipopolysaccharide (LPS) metabolite ADP-glycero-ß-D-manno-heptose (ADP-heptose), cytotoxin VacA, serine protease HtrA, and urease, and how they manipulate various key host cell signaling networks in the gastric epithelium. In particular, we highlight the H. pylori-induced disruption of cell-to-cell junctions, pro-inflammatory activities, as well as proliferative, pro-apoptotic and anti-apoptotic responses. Here we review these hijacked signal transduction events and their impact on gastric disease development.

CD248 induces a maladaptive unfolded protein response in diabetic kidney disease.

Dysfunction of mesangial cells plays a major role in the pathogenesis of diabetic kidney disease (DKD), the leading cause of kidney failure. However, the underlying molecular mechanisms are incompletely understood. By unbiased gene expression analysis of glucose-exposed mesangial cells, we identified the transmembrane receptor CD248 as the most upregulated gene, and the maladaptive unfolded protein response (UPR) as one of the most stimulated pathways. Upregulation of CD248 was further confirmed in glucose-stressed mesangial cells in vitro, in kidney glomeruli isolated from diabetic mice (streptozotocin; STZ and db/db models, representing type 1 and type 2 diabetes mellitus, respectively) in vivo, and in glomerular kidney sections from patients with DKD. Time course analysis revealed that glomerular CD248 induction precedes the onset of albuminuria, mesangial matrix expansion and maladaptive UPR activation (hallmarked by transcription factor C/EBP homologous protein (CHOP) induction) but is paralleled by loss of the adaptive UPR regulator spliced X box binding protein (XBP1). Mechanistically, CD248 promoted maladaptive UPR signaling via inhibition of the inositol requiring enzyme 1α (IRE1α)-mediated transcription factor XBP1 splicing in vivo and in vitro. CD248 induced a multiprotein complex comprising heat shock protein 90, BH3 interacting domain death agonist (BID) and IRE1α, in which BID impedes IRE1α-mediated XBP1 splicing and induced CHOP mediated maladaptive UPR signaling. While CD248 knockout ameliorated DKD-associated glomerular dysfunction and reverses maladaptive unfolded protein response signaling, concomitant XBP1 deficiency abolished the protective effect in diabetic CD248 knockout mice, supporting a functional interaction of CD248 and XBP1 in vivo. Hence, CD248 is a novel mesangial cell receptor inducing maladaptive UPR signaling in DKD.

Decreased eukaryotic initiation factors expression upon temozolomide treatment-potential novel implications for eIFs in glioma therapy.

PURPOSE: Since glioma therapy is currently still limited until today, new treatment options for this heterogeneous group of tumours are of great interest. Eukaryotic initiation factors (eIFs) are altered in various cancer entities, including gliomas. The purpose of our study was to evaluate the potential of eIFs as novel targets in glioma treatment. METHODS: We evaluated eIF protein expression and regulation in 22 glioblastoma patient-derived xenografts (GBM PDX) after treatment with established cytostatics and with regards to mutation profile analyses of GBM PDX. RESULTS: We observed decreased expression of several eIFs upon temozolomide (TMZ) treatment independent from the phosphatidylinositol 3-kinase (PI3K)/ AKT/ mammalian target of the rapamycin (mTOR) signalling pathway. These effects of TMZ treatment were not present in TMZ-resistant PDX. Combination therapy of regorafenib and TMZ re- established the eIF/AKT/mTOR axis. CONCLUSION: Our study provides novel insights into chemotherapeutic effects on eIF regulation in gliomas and suggests that eIFs are interesting candidates for future research to improve glioma therapy.

TIFA has dual functions in Helicobacter pylori-induced classical and alternative NF-κB pathways.

Helicobacter pylori infection constitutes one of the major risk factors for the development of gastric diseases including gastric cancer. The activation of nuclear factor-kappa-light-chain-enhancer of activated B cells (NF-κB) via classical and alternative pathways is a hallmark of H. pylori infection leading to inflammation in gastric epithelial cells. Tumor necrosis factor receptor-associated factor (TRAF)-interacting protein with forkhead-associated domain (TIFA) was previously suggested to trigger classical NF-κB activation, but its role in alternative NF-κB activation remains unexplored. Here, we identify TRAF6 and TRAF2 as binding partners of TIFA, contributing to the formation of TIFAsomes upon H. pylori infection. Importantly, the TIFA/TRAF6 interaction enables binding of TGFß-activated kinase 1 (TAK1), leading to the activation of classical NF-κB signaling, while the TIFA/TRAF2 interaction causes the transient displacement of cellular inhibitor of apoptosis 1 (cIAP1) from TRAF2, and proteasomal degradation of cIAP1, to facilitate the activation of the alternative NF-κB pathway. Our findings therefore establish a dual function of TIFA in the activation of classical and alternative NF-κB signaling in H. pylori-infected gastric epithelial cells.

Structural Dynamics of Lys11-Selective Deubiquitinylase Cezanne-1 during the Catalytic Cycle.

Deubiquitinylating enzymes (DUBs) regulate the deubiquitinylation process of post-translationally modified proteins and thus control protein signaling in various cellular processes. The DUB Cezanne-1 catalyzes the cleavage of the iso-peptide bond of Lys11-linked polyubiquitin chains with high selectivity. Crystal structures of Cezanne-1 in different states provide important insight regarding the complex formation and global changes during the catalytic cycle but are lacking details of dynamics and control of activation. Activity-based probes are used to isolate intermediate states upon forming covalent bonds with the DUB active site. Those, however, may lead to structures that are non-native. Conformational changes of Cezanne-1, during its process of activation and proteolytic activity, are investigated using all-atom molecular dynamics (MD) simulations of the ubiquitin-free, diubiquitin-bound, and monoubiquitin-bound Cezanne-1 DUB for a total of ∼18 µs. Our results show that ubiquitin-free Cezanne-1 dynamically shuttles between catalytically competent and incompetent states which suggests that its activation is independent of substrate binding. The catalytically competent substrate-free Cezanne-1 promotes distal ubiquitin substrate access to the catalytic center. The subsequent binding of the proximal ubiquitin shifts the equilibrium toward the catalytically competent state of the dyad, thereby promoting proteolysis of the iso-peptide bond. After cleavage of the scissile bond, sequential dissociation of first the proximal ubiquitin induces the inactivation of Cezanne-1. The subsequent release of the distal ubiquitin fully reconstitutes the inactive substrate-free state of Cezanne-1. The process of activation and catalytic turnover of DUB Cezanne-1 is a multistage cycle with several critical dynamic transitions that cannot be characterized based on protein structures alone. Activity-based probes of cysteine proteases lead to non-native protein-protein contacts, which need to be resolved in order to be able to issue statements about physiological states and substrate binding.

Monolithic hybrid abutment crowns (screw-retained) versus monolithic hybrid abutments with adhesively cemented monolithic crowns.

OBJECTIVES: The objective of this study is to compare monolithic hybrid abutment crowns (screw-retained) versus monolithic hybrid abutments with adhesively cemented monolithic single-tooth crowns. MATERIALS AND METHODS: Twenty subjects in need of an implant-borne restoration were randomly assigned to receive either a cement-retained (CRR) or a screw-retained (SRR) implant-supported monolithic lithium disilicate (LS2 ) reconstruction. Each patient received a titanium implant with in internal conic connection. After osseointegration and second-stage surgery, healing abutments were placed for about 10 days. The type of restoration (CRR vs. SRR) was randomly assigned, and the restorations were manufactured of monolithic LS2 . Both types of restorations, CRR and SRR, were based on a titanium component (Ti-base) that was bonded to the abutment (CRR) or the crown (SRR). The follow-up period for all restoration was 36 months. Clinical outcome was evaluated according to Functional Implant Prosthetic Score (FIPS). Quality of live (OHIP) and patient's satisfaction were assessed using patient-reported outcome measures (PROMs). Primary endpoint was loss of restoration for any reason. Kaplan-Meier curves were constructed and log-rank testing was performed (p < .05). RESULTS: One restoration of group CRR failed after 6 months due to loss of adhesion between Ti-base and individual abutment. No further biological or technical failures occurred. Kaplan-Meier analysis showed no significant difference between both treatment options (p = .317). There was no statistically significant difference between both types of restoration, neither for FIPS, OHIP, treatment time nor patient satisfaction (p > .05). CONCLUSION: Monolithic hybrid abutment crowns (screw-retained) and monolithic hybrid abutment with adhesively cemented monolithic crowns using lithium disilicate showed no statistically significant difference for implant-based reconstructions in this pilot RCT setting.

Helicobacter pylori-induced reactive oxygen species direct turnover of CSN-associated STAMBPL1 and augment apoptotic cell death.

Deubiquitinylases (DUBs) are central regulators of the ubiquitin system involved in protein regulation and cell signalling and are important for a variety of physiological processes. Most DUBs are cysteine proteases, and few other proteases are metalloproteases of the JAB1/MPN +/MOV34 protease family (JAMM). STAM-binding protein like 1 (STAMBPL1), a member of the JAMM family, cleaves ubiquitin bonds and has a function in regulating cell survival, Tax-mediated nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation and epithelial-mesenchymal transition. However, the molecular mechanism by which STAMBPL1 influences cell survival is not well defined, especially with regard to its deubiquitinylation function. Here, we show that reactive oxygen species (ROS) induced by chemotherapeutic agents or the human microbial pathogen Helicobacter pylori can induce cullin 1-RING ubiquitin ligase (CRL1) and 26S proteasome-dependent degradation STAMBPL1. Interestingly, STAMBPL1 has a direct interaction with the constitutive photomorphogenic 9 (COP9 or CSN) signalosome subunits CSN5 and CSN6. The interaction with the CSN is required for the stabilisation and function of the STAMBPL1 protein. In addition, STAMBPL1 deubiquitinylates the anti-apoptotic protein Survivin and thus ameliorates cell survival. In summary, our data reveal a previously unknown mechanism by which the deubiquitinylase STAMBPL1 and the E3 ligase CRL1 balance the level of Survivin degradation and thereby determine apoptotic cell death. In response to genotoxic stress, the degradation of STAMBPL1 augments apoptotic cell death. This new mechanism may be useful to develop therapeutic strategies targeting STAMBPL1 in tumours that have high STAMBPL1 and Survivin protein levels.

USP48 and A20 synergistically promote cell survival in Helicobacter pylori infection.

The human pathogen Helicobacter pylori represents a risk factor for the development of gastric diseases including cancer. The H. pylori-induced transcription factor nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) is involved in the pro-inflammatory response and cell survival in the gastric mucosa, and represents a trailblazer of gastric pathophysiology. Termination of nuclear NF-κB heterodimer RelA/p50 activity is regulated by the ubiquitin-RING-ligase complex elongin-cullin-suppressor of cytokine signalling 1 (ECSSOCS1), which leads to K48-ubiquitinylation and degradation of RelA. We found that deubiquitinylase (DUB) ubiquitin specific protease 48 (USP48), which interacts with the COP9 signalosome (CSN) subunit CSN1, stabilises RelA by deubiquitinylation and thereby promotes the transcriptional activity of RelA to prolong de novo synthesis of DUB A20 in H. pylori infection. An important role of A20 is the suppression of caspase-8 activity and apoptotic cell death. USP48 thus enhances the activity of A20 to reduce apoptotic cell death in cells infected with H. pylori. Our results, therefore, define a synergistic mechanism by which USP48 and A20 regulate RelA and apoptotic cell death in H. pylori infection.

Synthesis and structure-activity relationships of USP48 deubiquitinylase inhibitors.

Ubiquitin-specific proteases represent a family of enzymes that catalyze the cleavage of ubiquitin from specific substrate proteins to regulate their activity. USP48 is a rarely studied USP, which has recently been linked to inflammatory signaling via regulation of the transcription factor nuclear factor kappa B. Nonetheless, a crystal structure of USP48 has not yet been resolved and potent inhibitors are not known. We screened a set of 14 commercially available USP inhibitors for their activity against USP48 and identified the USP2 inhibitor "ML364" as a candidate for further optimization. Using a ligand-based approach, we derived and synthesized a series of ML364 analogs. The IC50 concentrations of the new compounds to inhibit USP48 were determined in a deubiquitinylase activity assay by measuring the fluorescence intensity using tetra-ubiquitin rhodamine110 as substrate. A compound containing a carboxylic acid functionalization (17e) inhibited USP48 activity toward tetra-ubiquitin rhodamine110 with an IC50 of 12.6 µM. Further structure-based refinements are required to improve the inhibition activity and specificity.

Health economic evaluation of forced orthodontic extrusion of extensively damaged teeth: up to 6-year results from a clinical study.

OBJECTIVES: Clinical data on retaining extensively damaged teeth using forced orthodontic extrusion followed by restorative rehabilitation are scarce, and economic evaluations are basically absent. A health economic evaluation of this method was performed based on a clinical study. MATERIALS AND METHODS: In a convenience sample of individuals recruited from routine care, extensively damaged teeth were orthodontically extruded prior to restoration. Patients were followed up for up to 6 years. The health outcome was tooth retention time. Direct medical, non-medical, and indirect initial and follow-up costs were estimated using the private payer's perspective in German healthcare. Association of initial direct medical treatment costs and cofounding variables was analyzed using generalized linear models. Success and survival were secondary outcomes. RESULTS: A total of 35 teeth in 30 patients were followed over a mean ± SD of 49 ± 19 months. Five patients (14%) dropped out during that period. Median initial costs were 1941€ (range: 1284-4392€), median costs for follow-up appointments were 215€ (range: 0-5812€), and median total costs were 2284€ (range: 1453 to 7109€). Endodontic re-treatment and placement of a post had a significant impact on total costs. Three teeth had to be extracted and in three patients orthodontic relapse was observed. The survival and success rates were 91% and 83%, respectively. CONCLUSIONS: Within the limitations of this clinical study, total treatment costs for orthodontic extrusion and subsequent restoration of extensively damaged teeth were considerable. Costs were by large generated initially; endodontic and post-endodontic therapies were main drivers. Costs for retreatments due to complications were limited, as only few complications arose. CLINICAL RELEVANCE: The restoration of extensively damaged teeth after forced orthodontic extrusion comes with considerable initial treatment costs, but low follow-up costs. Overall and over the observational period and within German healthcare, costs are below those for tooth replacement using implant-supported crowns. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: DRK S00026697).

Root-analogue implants compared to forced orthodontic extrusion: a retrospective analysis of clinical, radiological and esthetic outcomes after restoration.

OBJECTIVES: To assess clinical, radiological and esthetic outcomes of restorations supported by root-analogue implants (RAIs) or roots of severely damaged teeth after forced orthodontic extrusion (FOE). MATERIALS AND METHODS: Clinical data regarding milled one-piece (titanium/zirconia roots and zirconia abutments) RAIs (REPLICATE™ System) and FOE were recorded and retrospectively evaluated for 40 patients by two investigators. Strict inclusion and exclusion criteria were applied. Functional and esthetic outcomes were assessed for n = 20 pre-molars and n = 20 anterior teeth via comparison of radiographic and digital images applying the novel Functional Implant Prosthodontic Score (FIPS). Krippendorff's alpha coefficient was calculated to assess inter-rater reliability. Mann-Whitney-U-Test was used to compare the assessed parameters. Level of significance was set to p < 0.05. RESULTS: After a mean observation period of 18.4 ± 5.7 months for restorations supported by RAIs and 43.9 ± 16.4 months for restorations after FOE, mean FIPS scores were 9.2/8.8 ± 1.1/1.2 (RAIs) and 7.4/7.7 ± 1.3/1.5 (FOE), respectively. Krippendorff's alpha coefficients did not reveal unacceptable inter-rater reliabilities regarding the investigators and applicability of FIPS. Significant differences were documented when comparing restorations after FOE or supported by RAIs regarding bone loss (p < 0.01), presence of papillae (p < 0.05) and quality and quantity of mucosa (p < 0.02) in favor of FOE. CONCLUSIONS: Within the main limitations of sample size and the retrospective study design, both concepts seem to provide clinically acceptable results. CLINICAL RELEVANCE: Bone- and tissue-preserving characteristics regarding the concept of FOE are promising. It could be applicable for socket preservation and subsequent conventional implant placements in an adapted workflow.

Forced orthodontic extrusion to restore extensively damaged anterior and premolar teeth as abutments for single-crown restorations: Up to 5-year results from a pilot clinical study.

STATEMENT OF PROBLEM: Clinical data on orthodontic extrusion to restore teeth deemed unrestorable because of their defect size are scarce. It remains unclear for which defects forced orthodontic extrusion and tooth retention is preferred to extraction. PURPOSE: The purpose of this pilot clinical study was to investigate the survival, frequency, and type of complications of extensively damaged teeth requiring single-crown restorations after forced orthodontic extrusion. MATERIAL AND METHODS: Participants were recruited from consecutive patients in need of restorative treatment of extensively damaged teeth at a university clinic. The teeth were orthodontically extruded to reestablish the biologic width and to ensure a 2-mm ferrule preparation before restoration. The primary endpoint was restoration success and survival. At recall, survival was defined as the tooth being in situ and success as a symptom-free tooth with an intact, caries-free restoration and with physiological pocket probing depths, no signs of intrusion, ankylosis, root resorption, or periapical radiolucency. Recalls were performed every 6 months; the outcome was assessed by radiographic and clinical evaluation after up to 5 years of clinical service. Quantitative parameters were described with mean values and standard deviations. RESULTS: Thirty-four participants were assessed for eligibility and enrolled (mean ±standard deviation age: 53.4 ±18.9 years). Four participants were premature dropouts. Data were analyzed for 35 teeth in 30 participants. The amount of extrusion varied between 2 and 6 mm (mean ±standard deviation 3.4 ±0.9 mm). The mean duration of extrusion was 18.9 ±12.6 days and the mean duration of retention was 126.94 ±88.1 days. The mean ±standard deviation crown-to-root ratio after treatment was 0.8 ±0.1 (range: 0.5 to 1.0). Three participants exhibited orthodontic relapse before restoration. Teeth were successfully restored after repeated extrusion. After a mean observation period of 3.3 years (range: 1 to 5.2 years), 29 of 31 teeth were still in situ. Two teeth were fractured, and 4 participants were not available for recall. Thus, the survival rate was 94%. No resorption or periapical translucencies were observed radiographically. Clinical examinations revealed physiological probing depths and absence of ankyloses. One tooth presented with marginal bone loss. The most frequent type of complication was orthodontic relapse at recall (n=3). A total of 84% of teeth were considered a success. CONCLUSIONS: Forced orthodontic extrusion allowed for the restoration of anterior and premolar teeth deemed as nonrestorable because of their defect size. Tooth retention of extensively damaged teeth and their use as abutments for single-crown restorations can be recommended.

Cullin 3-Based Ubiquitin Ligases as Master Regulators of Mammalian Cell Differentiation.

Specificity of the ubiquitin proteasome system is controlled by ubiquitin E3 ligases, including their major representatives, the multisubunit cullin-RING ubiquitin (Ub) ligases (CRLs). More than 200 different CRLs are divided into seven families according to their cullin scaffolding proteins (CUL1-7) around which they are assembled. Research over two decades has revealed that different CRL families are specialized to fulfill specific cellular functions. Whereas many CUL1-based CRLs (CRL1s) ubiquitylate cell cycle regulators, CRL4 complexes often associate with chromatin to control DNA metabolism. Based on studies about differentiation programs of mesenchymal stem cells (MSCs), including myogenesis, neurogenesis, chondrogenesis, osteogenesis and adipogenesis, we propose here that CRL3 complexes evolved to fulfill a pivotal role in mammalian cell differentiation.

Substrate-assisted activation and selectivity of the bacterial RavD effector deubiquitinylase.

Deubiquitinylases (DUBs) catalyze the peptide bond cleavage of specific ubiquitin linkages at distinct protein substrates. Pathogens from viruses and bacteria independently developed effector proteins with DUB activity to mimic host DUB functions and circumvent immune responses. The effector protein RavD from Legionella pneumophila cleaves linear ubiquitin chains with an exclusive methionine-1 selectivity. It thus performs as a functional analogue of the human DUB OTULIN, which achieves its selectivity only via a specialized proximal ubiquitin S1' binding site as well as a substrate-assisted activation of the catalytic triad. An analysis of the crystal structures of bacterial RavD in its free and di-ubiquitin-bound forms, in order to rationalize the structural basis for its selectivity and activation mechanism, is not fully conclusive. As these ambiguities might arise from the introduced double mutation of the di-ubiquitin substrate in the RavD-di-ubiquitin complex crystal structure, biomolecular modeling, and molecular dynamics sampling (1-2 µs for each system of RavD and OTULIN) were employed to reconstitute the physiological RavD-di-ubiquitin complex. The simulations show that the distal S1 ubiquitin binding sites of RavD and OTULIN are similar in terms of interface area, composition, and ubiquitin binding affinity. The proximal S1' site of RavD, in contrast, is significantly smaller and ubiquitin binding is weaker and more flexible than in OTULIN. Upon substrate access, the residues of the catalytic triad of RavD show a reduction of flexibility and a conformational transition toward a catalytically active state. Thus, the enzymatic activation of RavD is presumably also substrate-assisted and a clear rationale for the common M1-substrate selectivity.

Peptide microarray-based analysis of antibody responses to SARS-CoV-2 identifies unique epitopes with potential for diagnostic test development.

Humoral immunity to the Severe Adult Respiratory Syndrome (SARS) Coronavirus (CoV)-2 is not fully understood yet but is a crucial factor of immune protection. The possibility of antibody cross-reactivity between SARS-CoV-2 and other human coronaviruses (HCoVs) would have important implications for immune protection but also for the development of specific diagnostic ELISA tests. Using peptide microarrays, n = 24 patient samples and n = 12 control samples were screened for antibodies against the entire SARS-CoV-2 proteome as well as the Spike (S), Nucleocapsid (N), VME1 (V), R1ab, and Protein 3a (AP3A) of the HCoV strains SARS, MERS, OC43, and 229E. While widespread cross-reactivity was revealed across several immunodominant regions of S and N, IgG binding to several SARS-CoV-2-derived peptides provided statistically significant discrimination between COVID-19 patients and controls. Selected target peptides may serve as capture antigens for future, highly COVID-19-specific diagnostic antibody tests.

Manifold role of ubiquitin in Helicobacter pylori infection and gastric cancer.

Infection with H. pylori induces a strong host cellular response represented by induction of a set of molecular signaling pathways, expression of proinflammatory cytokines and changes in proliferation. Chronic infection and inflammation accompanied by secretory dysfunction can result in the development of gastric metaplasia and gastric cancer. Currently, it has been determined that the regulation of many cellular processes involves ubiquitinylation of molecular effectors. The binding of ubiquitin allows the substrate to undergo a change in function, to interact within multimolecular signaling complexes and/or to be degraded. Dysregulation of the ubiquitinylation machinery contributes to several pathologies, including cancer. It is not understood in detail how H. pylori impacts the ubiquitinylation of host substrate proteins. The aim of this review is to summarize the existing literature in this field, with an emphasis on the role of E3 ubiquitin ligases in host cell homeodynamics, gastric pathophysiology and gastric cancer.

Matrix Metalloproteinases in Helicobacter pylori-Associated Gastritis and Gastric Cancer.

Gastric cancer is one of the leading causes of the cancer-related mortality worldwide. The etiology of this disease is complex and involves genetic predisposition and environmental factors, including Helicobacter pylori. Infection of the stomach with H. pylori leads to gastritis and gastric atrophy, which can progress stepwise to gastric cancer. Matrix metalloproteinases (MMPs) actively participate in the pathology development. The further progression of gastric cancer seems to be less dependent on bacteria but of intra-tumor cell dynamics. Bioinformatics data confirmed an important role of the extracellular matrix constituents and specific MMPs in stomach carcinoma invasion and metastasis, and revised their potential as predictors of the disease outcome. In this review, we describe, in detail, the impact of MMPs in H. pylori-associated gastritis and gastric cancer.