Ultrastructural localization of 5-methylcytosine on DNA and RNA

DNA methylation is the major epigenetic modification and it is involved in the negative regulation of gene expression. Its alteration can lead to neoplastic transformation. Several biomolecular approaches are nowadays used to study this modification on DNA, but also on RNA molecules, which are known to play a role in different biological processes. RNA methylation is one of the most common RNA modifications and 5-methylcytosine presence has recently been suggested in mRNA. However, an analysis of nucleic acid methylation at electron microscope is still lacking. Therefore, we visualized DNA methylation status and RNA methylation sites in the interphase nucleus of HeLa cells and rat hepatocytes by ultrastructural immunocytochemistry and cytochemical staining. This approach represents an efficient alternative to study nucleic acid methylation. In particular, this ultrastructural method makes the visualization of this epigenetic modification on a single RNA molecule possible, thus overcoming the technical limitations for a (pre-)mRNA methylation analysis.     

Deciphering the BAR code of membrane modulators

The BAR domain is the eponymous domain of the “BAR-domain protein superfamily”, a large and diverse set of mostly multi-domain proteins that play eminent roles at the membrane cytoskeleton interface. BAR domain homodimers are the functional units that peripherally associate with lipid membranes and are involved in membrane sculpting activities. Differences in their intrinsic curvatures and lipid-binding properties account for a large variety in membrane modulating properties. Membrane activities of BAR domains are further modified and regulated by intramolecular or inter-subunit domains, by intermolecular protein interactions, and by posttranslational modifications. Rather than providing detailed cell biological information on single members of this superfamily, this review focuses on biochemical, biophysical, and structural aspects and on recent findings that paradigmatically promote our understanding of processes driven and modulated by BAR domains.     

Development of liquid-nitrogen-cooling friction stir spot welding for AZ31 magnesium alloy joints

A liquid-nitrogen-cooling friction stir spot welding (C-FSSW) technology was developed for welding AZ31 magnesium alloy sheets. The liquid-nitrogen cooling degraded the deformability of the welded materials such that the width of interfacial cracks increased with increasing cooling time. The grain size of the stirred zone (SZ) and the heat-affected zone (HAZ) of the C-FSSW-welded joints decreased, whereas that of the thermomechanically affected zone (TMAZ) increased with increasing cooling time. The maximum tensile shear load of the C-FSSW-welded joints welded with a cooling time of 5 or 7 s was larger than that of the friction stir spot welding (FSSW)-welded joint, and the tensile shear load decreased with increasing cooling time. The microhardness of the C-FSSW-welded joints was greater than that of the FSSW-welded joint. Moreover, the microhardness of the SZ and the HAZ of the C-FSSW-welded joints increased, whereas that of the TMAZ decreased, with increasing cooling time.     

Studies on induction hardening of powder-metallurgy-processed Fe-Cr/Mo alloys

Induction hardening of dense Fe-Cr/Mo alloys processed via the powder-metallurgy route was studied. The Fe-3Cr-0.5Mo, Fe-1.5Cr-0.2Mo, and Fe-0.85Mo pre-alloyed powders were mixed with 0.4wt%, 0.6wt%, and 0.8wt% C and compacted at 500, 600, and 700 MPa, respectively. The compacts were sintered at 1473 K for 1 h and then cooled at 6 K/min. Ferrite with pearlite was mostly observed in the sintered alloys with 0.4wt% C, whereas a carbide network was also present in the alloys with 0.8wt% C. Graphite at prior particle boundaries led to deterioration of the mechanical properties of alloys with 0.8wt% C, whereas no significant induction hardening was achieved in alloys with 0.4wt% C. Among the investigated samples, alloys with 0.6wt% C exhibited the highest strength and ductility and were found to be suitable for induction hardening. The hardening was carried out at a frequency of 2.0 kHz for 2-3 s. A case depth of 2.5 mm was achieved while maintaining the bulk (interior) hardness of approximately HV 230. A martensitic structure was observed on the outer periphery of the samples. The hardness varied from HV 600 to HV 375 from the sample surface to the interior of the case hardened region. The best combination of properties and hardening depth was achieved in case of the Fe-1.5Cr-0.2Mo alloy with 0.6wt% C.     

Using the ‘Engagement’ Model of Problem Solving to Assist Students in Capstone Learning

The engagement model for problem solving was developed as a response to soft systems methodology being difficult to implement in certain circumstances. In the years since the original paper was published, the model has developed into an iterative problem solving approach for the assessment, creation and implementation of new ideas in difficult problem situations. This case study paper exemplifies the kind of work undertaken through the use of the model and provides a practical scenario for how one might apply it. The case study takes place in a higher education environment and relates to the development of a capstone course for management majors. The paper concludes with suggestions for future research.     

Control of DNA integrity in skeletal muscle under physiological and pathological conditions

Skeletal muscle is a highly oxygen-consuming tissue that ensures body support and movement, as well as nutrient and temperature regulation. DNA damage induced by reactive oxygen species is present in muscles and tends to accumulate with age. Here, we present a summary of data obtained on DNA damage and its implication in muscle homeostasis, myogenic differentiation and neuromuscular disorders. Controlled and transient DNA damage appears to be essential for muscular homeostasis and differentiation while uncontrolled and chronic DNA damage negatively affects muscle health.     

Opposing effects on the cell cycle of T lymphocytes by Fbxo7 via Cdk6 and p27

G₁ phase cell cycle proteins, such as cyclin-dependent kinase 6 (Cdk6) and its activating partners, the D-type cyclins, are important regulators of T-cell development and function. An F-box protein, called F-box only protein 7 (Fbxo7), acts as a cell cycle regulator by enhancing cyclin D-Cdk6 complex formation and stabilising levels of p27, a cyclin-dependent kinase inhibitor. We generated a murine model of reduced Fbxo7 expression to test its physiological role in multiple tissues and found that these mice displayed a pronounced thymic hypoplasia. Further analysis revealed that Fbxo7 differentially affected proliferation and apoptosis of thymocytes at various stages of differentiation in the thymus and also mature T-cell function and proliferation in the periphery. Paradoxically, Fbxo7-deficient immature thymocytes failed to undergo expansion in the thymus due to a lack of Cdk6 activity, while mature T cells showed enhanced proliferative capacity upon T-cell receptor engagement due to reduced p27 levels. Our studies reveal differential cell cycle regulation by Fbxo7 at different stages in T-cell development.     

Melatonin transport into mitochondria

Melatonin is a well-known, nighttime-produced indole found in bacteria, eukaryotic unicellulars, animals or vascular plants. In vertebrates, melatonin is the major product of the pineal gland, which accounts for its increase in serum during the dark phase, but it is also produced by many other organs and cell types. Such a wide distribution is consistent with its multiple and well-described functions which include from the circadian regulation and adaptation to seasonal variations to immunomodulatory and oncostatic actions in different types of tumors. The discovery of its antioxidant properties in the early 1990s opened a new field of potential protective functions in multiple tissues. A special mention should be made regarding the nervous system, where the indole is considered a major neuroprotector. Furthermore, mitochondria appear as one of the most important targets for the indole’s protective actions. Melatonin’s mechanisms of action vary from the direct molecular interaction with free radicals (free radical scavenger) to the binding to membrane (MLT1A and MLT1B) or nuclear receptors (RZR/RORα). Receptor binding has been associated with some, but not all of the indole functions reported to date. Recently, two new mechanisms of cellular uptake involving the facilitative glucose transporters GLUT/SLC2A and the proton-driven oligopeptide transporter PEPT1/2 have been reported. Here we discuss the potential importance that these newly discovered transport systems could have in determining the actions of melatonin, particularly in the mitochondria. We also argue the relative importance of passive diffusion vs active transport in different parts of the cell.     

System optimization model of adoption of a new infrastructure with multi-resource and multi-demand sites

This study develops a conceptual system optimization model of adoption of a new infrastructure technology with multiple resource sites and multiple demand sites. With the model, this paper analyzes how the distance, spillover effect, demand, initial investment cost, and learning rate influence the adoption of the new infrastructure technology and presents optimization results of the model in different scenarios. The main findings of the study are: from the perspective of system optimization, (1) different distances among different resource-demand pairs will result in different adoption time of a new infrastructure; (2) technological spillover among different resource-demand pairs will accelerate the adoption of a new infrastructure; (3) it is hard to say that higher demand will pull faster adoption of a new infrastructure, and the optimal time of adopting of a new infrastructure is very sensitive to its technological learning rate.     

Social Media as a Mean for Improved Technical Communication

Technical communication (TC) is an important activity in order to provide the users of technical artifacts with necessary information concerning the operation and maintenance of the products they are using. In the current working procedures of TC, however, the users become too passive. They are just receivers of information. Further, there is no effective feedback from the users to the producers of the information. In order to overcome those problems we here propose an application of social media (SM) in future TC. By using SM as a complementary channel for TC it becomes possible for technology firms to boost their competitive advantage and to improve the quality and completeness of their TC. This technique, however, is still more of a potential and less of a reality. Several problems, mostly concerning security and confidentiality remain to be solved. The empirical base for this work comes from TIC, an EU-funded development project involving several Swedish technology companies. Published research results are somewhat scattered but several sources coherently indicate both the need and potential for SM as a vehicle in TC. By scanning the net it is further possible to identify a number of seemingly successful applications of SM in such applications.