Recovery of protein synthesis to assay DNA repair activity in transcribed genes in living cells and tissues

Transcription-coupled nucleotide excision repair (TC-NER) is an important DNA repair mechanism that protects against the negative effects of transcription-blocking DNA lesions. Hereditary TC-NER deficiencies cause pleiotropic and often severe neurodegenerative and progeroid symptoms. While multiple assays have been developed to determine TC-NER activity for clinical and research purposes, monitoring TC-NER is hampered by the low frequency of repair events occurring in transcribed DNA. ’Recovery of RNA Synthesis’ is widely used as indirect TC-NER assay based on the notion that lesion-blocked transcription only resumes after successful TC-NER. Here, we show that measuring novel synthesis of a protein after its compound-induced degradation prior to DNA damage induction is an equally effective but more versatile manner to indirectly monitor DNA repair activity in transcribed genes. This ‘Recovery of Protein Synthesis’ (RPS) assay can be adapted to various degradable proteins and readouts, including imaging and immunoblotting. Moreover, RPS allows real-time monitoring of TC-NER activity in various living cells types and even in differentiated tissues of living organisms. To illustrate its utility, we show that DNA repair in transcribed genes declines in aging muscle tissue of C. elegans. Therefore, the RPS assay constitutes an important novel clinical and research tool to investigate transcription-coupled DNA repair

Cross-National Logo Evaluation Analysis: An Individual Level Approach

The universality of design perception and response is tested using data collected from ten countries: Argentina, Australia, China, Germany, Great Britain, India, the Netherlands, Russia, Singapore, and the United States. A Bayesian, finite-mixture, structural-equation model is developed that identifies latent logo clusters while accounting for heterogeneity in evaluations. The concomitant variable approach allows cluster probabilities to be country specific. Rather than a priori defined clusters, our procedure provides a posteriori cross-national logo clusters based on consumer response similarity. To compare the a posteriori cross-national logo clusters, our approach is integrated with Steenkamp and Baumgartner’s (1998) measurement invariance methodology. Our model reduces the ten countries to three cross-national clusters that respond differently to logo design dimensions: the West, Asia, and Russia. The dimensions underlying design are found to be similar across countries, suggesting that elaborateness, naturalness, and harmony are universal design dimensions. Responses (affect, shared meaning, subjective familiarity, and true and false recognition) to logo design dimensions (elaborateness, naturalness, and harmony) and elements (repetition, proportion, and parallelism) are also relatively consistent, although we find minor differences across clusters. Our results suggest that managers can implement a global logo strategy, but they also can optimize logos for specific countries if desired.adaptation;standardization;Bayesian;international marketing;design;Gibbs sampling;concomitant variable;logos;mixture models;structural equation models

The Multidimensional Study of Viral Campaigns as Branching Processes

Viral campaigns on the Internet may follow variety of models, depending on the content, incentives, personal attitudes of sender and recipient to the content and other factors. Due to the fact that the knowledge of the campaign specifics is essential for the campaign managers, researchers are constantly evaluating models and real-world data. The goal of this article is to present the new knowledge obtained from studying two viral campaigns that took place in a virtual world which followed the branching process. The results show that it is possible to reduce the time needed to estimate the model parameters of the campaign and, moreover, some important aspects of time-generations relationship are presented.Comment: In proceedings of the 4th International Conference on Social Informatics, SocInfo 201

After Nerve Injury, Lineage Tracing Shows That Myelin and Remak Schwann Cells Elongate Extensively and Branch to Form Repair Schwann Cells, Which Shorten Radically on Remyelination

There is consensus that, distal to peripheral nerve injury, myelin and Remak cells reorganize to form cellular columns, Bungner's bands, which are indispensable for regeneration. However, knowledge of the structure of these regeneration tracks has not advanced for decades and the structure of the cells that form them, denervated or repair Schwann cells, remains obscure. Furthermore, the origin of these cells from myelin and Remak cells and their ability to give rise to myelin cells after regeneration has not been demonstrated directly, although these conversions are believed to be central to nerve repair. Using genetic lineage-tracing and scanning-block face electron microscopy, we show that injury of sciatic nerves from mice of either sex triggers extensive and unexpected Schwann cell elongation and branching to form long, parallel processes. Repair cells are 2- to 3-fold longer than myelin and Remak cells and 7- to 10-fold longer than immature Schwann cells. Remarkably, when repair cells transit back to myelinating cells, they shorten ∼7-fold to generate the typically short internodes of regenerated nerves. The present experiments define novel morphological transitions in injured nerves and show that repair Schwann cells have a cell-type-specific structure that differentiates them from other cells in the Schwann cell lineage. They also provide the first direct evidence using genetic lineage tracing for two basic assumptions in Schwann cell biology: that myelin and Remak cells generate the elongated cells that build Bungner bands in injured nerves and that such cells can transform to myelin cells after regeneration

Studying Paths of Participation in Viral Diffusion Process

Authors propose a conceptual model of participation in viral diffusion process composed of four stages: awareness, infection, engagement and action. To verify the model it has been applied and studied in the virtual social chat environment settings. The study investigates the behavioral paths of actions that reflect the stages of participation in the diffusion and presents shortcuts, that lead to the final action, i.e. the attendance in a virtual event. The results show that the participation in each stage of the process increases the probability of reaching the final action. Nevertheless, the majority of users involved in the virtual event did not go through each stage of the process but followed the shortcuts. That suggests that the viral diffusion process is not necessarily a linear sequence of human actions but rather a dynamic system.Comment: In proceedings of the 4th International Conference on Social Informatics, SocInfo 201

Incidence and Functional Outcomes of Scapholunate Diastases Associated Distal Radius Fractures: A 2-year Follow-Up Scapholunate Dissociation

Background: The Scapholunate Interosseous Ligament (SLIL) is the first intrinsic carpal ligament to be injured in wrist trauma, present in up to 64% of the distal radius fractures. However, it remains unclear what patients develop symptoms, making primary treatment of these injuries accompanying distal radius fractures remains questionable. Objective: The aim of this study was to evaluate the functional outcomes of patients with scapholunate diastasis associated with distal radius fractures. Methods: We evaluated 391 patients with a distal radius fracture. Using Computer Tomography (CT) scans the scapholunate interval was measured. We identified 14 patients with an SLD (>3mm) of the injured wrist, which underwent a CT-scan of the contralateral wrist. To evaluate the functional outcomes at a mean follow up of 136±90 weeks, we used the Quick Disabilities of the Arm, Shoulder and Hand (qDASH) Score. Results: There were 8 patients with bilateral SLD and 6 patients with unilateral SLD. Five patients had a qDASH score of 0 and one patient showed a qDASH score of 18.2. The patient with a poor score had bilateral preexisting osteoarthritis of the wrist. No patient had additional surgery of the SLIL. Conclusion: In patients with distal radius fractures, more than half of the 14 patients with an SL gap on CT had widening on the contralateral side. It is therefore worthwhile to image the contralateral wrist before diagnosing a SLD. The patients with unilateral SLD should not be surgically treated at initial presentation because they may have good functional outcomes after a follow up of 2 years

Groene diensten in nationale landschappen : potenties bij een veranderende landbouw

De landbouw is voor een belangrijk deel drager van het landschap in Nationale Landschappen. Dit rapport verkent de betekenis van groene diensten door de landbouw, tegen de achtergrond van verschillende ontwikkelingsrichtingen in die landbouw (schaalvergroting - verbreding). Twee kwalitatieve visies worden ontwikkeld aan de hand van al bestaande scenariostudies (de zogenaamde CPB-scenario’s), een analyse van instrumenten van groene diensten in nationaal en internationaal verband en een discussiemiddag. Beide visies laten ontwikkelingsrichtingen van Nationale landschappen zien en werken de rol van groene diensten door de landbouw verder uit. Deze rol wordt uitgewerkt aan de hand van een raamwerk over de organisatie van groene diensten en een analyse van de te realiseren kernkwaliteiten in Nationale Landschappen

G Protein-Coupled Receptor Kinase Function Is Essential for Chemosensation in C. elegans

AbstractG protein-coupled receptors (GPCRs) mediate diverse signaling processes, including olfaction. G protein-coupled receptor kinases (GRKs) are important regulators of G protein signal transduction that specifically phosphorylate activated GPCRs to terminate signaling. Despite previously described roles for GRKs in GPCR signal downregulation, animals lacking C. elegans G protein-coupled receptor kinase-2 (Ce-grk-2) function are not hypersensitive to odorants. Instead, decreased Ce-grk-2 function in adult sensory neurons profoundly disrupts chemosensation, based on both behavioral analysis and Ca2+ imaging. Although mammalian arrestin proteins cooperate with GRKs in receptor desensitization, loss of C. elegans arrestin-1 (arr-1) does not disrupt chemosensation. Either overexpression of the C. elegans Gα subunit odr-3 or loss of eat-16, which encodes a regulator of G protein signaling (RGS) protein, restores chemosensation in Ce-grk-2 mutants. These results demonstrate that loss of GRK function can lead to reduced GPCR signal transduction and suggest an important role for RGS proteins in the regulation of chemosensation

Soft-tissue coverage for wound complications following total elbow arthroplasty

Background In patients with total elbow arthroplasty (TEA), the soft-tissue around the elbow can be vulnerable to soft-tissue complications. This study aims to assess the outcomes after soft-tissue reconstruction following TEA. Methods We retrospectively included nine adult patients who underwent soft-tissue reconstruction following TEA. Demographic data and disease characteristics were collected through medical chart reviews. Additionally, we contacted all four patients that were alive at the time of the study by phone to assess any current elbow complications. Local tissue rearrangement was used for soft-tissue reconstruction in six patients, and a pedicle flap was used in three patients. The median follow-up period was 1.3 years (range, 6 months–14.7 years). Results Seven patients (78%) underwent reoperation. Four patients (44%) had a reoperation for soft-tissue complications, including dehiscence or nonhealing of infected wounds. Five patients (56%) had a reoperation for implant-related complications, including three infections and two peri-prosthetic fractures. At the final follow-ups, six patients (67%) achieved successful wound healing and two patients had continued wound healing issues, while two patients had an antibiotic spacer in situ and one patient underwent an above-the-elbow amputation. Conclusions This study reports a complication rate of 78% for soft-tissue reconstructions after TEA. Successful soft-tissue healing was achieved in 67% of patients, but at the cost of multiple surgeries. Early definitive soft-tissue reconstruction could prove to be preferable to minor interventions such as irrigation, debridement, and local tissue advancement, or smaller soft-tissue reconstructions using local tissue rearrangement or a pedicled flap at a later stage