Supramolecular Assembly of Aminoethylene‐Lipopeptide PMO Conjugates into RNA Splice‐Switching Nanomicelles

Phosphorodiamidate morpholino oligomers (PMOs) are oligonucleotide analogs that can be used for therapeutic modulation of pre‐mRNA splicing. Similar to other classes of nucleic acid‐based therapeutics, PMOs require delivery systems for efficient transport to the intracellular target sites. Here, artificial peptides based on the oligo(ethylenamino) acid succinyl‐tetraethylenpentamine (Stp), hydrophobic modifications, and an azide group are presented, which are used for strain‐promoted azide‐alkyne cycloaddition conjugation with splice‐switching PMOs. By systematically varying the lead structure and formulation, it is determined that the type of contained fatty acid and supramolecular assembly have a critical impact on the delivery efficacy. A compound containing linolenic acid with three cis double bonds exhibits the highest splice‐switching activity and significantly increases functional protein expression in pLuc/705 reporter cells in vitro and after local administration in vivo. Structural and mechanistic studies reveal that the lipopeptide PMO conjugates form nanoparticles, which accelerate cellular uptake and that the content of unsaturated fatty acids enhances endosomal escape. In an in vitro Duchenne muscular dystrophy exon skipping model using H2K‐mdx52 dystrophic skeletal myotubes, the highly potent PMO conjugates mediate significant splice‐switching at very low nanomolar concentrations. The presented aminoethylene‐lipopeptides are thus a promising platform for the generation of PMO‐therapeutics with a favorable activity/toxicity profile.     

Antenna phase pattern measurements at millimeter wave frequencies using the differential phase method with only one power meter

The differential phase measurement method has been improved to need the use of only one power meter instead of three power meters. This enables accurate antenna phase pattern measurements with a simplified set-up, accompanied by the reduction in cost. All advantages of the differential phase measurement method are still also available, e.g. there is no need to phase lock oscillators or to use rotary joints. The measurement results for an antenna at 110 GHz are presented. A good agreement with earlier data was obtained.     

A signal-detection theory based perspective on design of warning

The effects of warnings are analyzed using a distributed signal-detection theory model. It is established that selectivity always increases effectiveness. The implications to optimal warning design for intermittent versus continuous hazards are discussed. The changes in the behavior of the 6 human subjects in response to changes in the warning levels are consistent with the predictions of the model.     

Expression and Roles of Individual HIF Prolyl 4-Hydroxylase Isoenzymes in the Regulation of the Hypoxia Response Pathway along the Murine Gastrointestinal Epithelium

The HIF prolyl 4-hydroxylases (HIF-P4H) control hypoxia-inducible factor (HIF), a powerful mechanism regulating cellular adaptation to decreased oxygenation. The gastrointestinal epithelium subsists in “physiological hypoxia” and should therefore have an especially well-designed control over this adaptation. Thus, we assessed the absolute mRNA expression levels of the HIF pathway components, Hif1a, HIF2a, Hif-p4h-1, 2 and 3 and factor inhibiting HIF (Fih1) in murine jejunum, caecum and colon epithelium using droplet digital PCR. We found a higher expression of all these genes towards the distal end of the gastrointestinal tract. We detected mRNA for Hif-p4h-1, 2 and 3 in all parts of the gastrointestinal tract. Hif-p4h-2 had significantly higher expression levels compared to Hif-p4h-1 and 3 in colon and caecum epithelium. To test the roles each HIF-P4H isoform plays in the gut epithelium, we measured the gene expression of classical HIF target genes in Hif-p4h-1^−/−, Hif-p4h-2 hypomorph and Hif-p4h-3^−/− mice. Only Hif-p4h-2 hypomorphism led to an upregulation of HIF target genes, confirming a predominant role of HIF-P4H-2. However, the abundance of Hif-p4h-1 and 3 expression in the gastrointestinal epithelium implies that these isoforms may have specific functions as well. Thus, the development of selective inhibitors might be useful for diverging therapeutic needs.     

Fault tolerant machine learning for nanoscale cognitive radio

We introduce a machine learning-based classifier that identifies free radio channels for cognitive radio. The architecture is designed for nanoscale implementation, under nanoscale implementation constraints; we do not describe all physical details but believe future physical implementation to be feasible. The system uses analog computation and consists of cyclostationary feature extraction and a radial basis function network for classification. We describe a model for nanoscale faults in the system, and simulate experimental performance and fault tolerance in recognizing WLAN signals, under different levels of noise and computational errors. The system performs well under expected non-ideal manufacturing and operating conditions.     

Semiautomated Identification and Classification of Customer Complaints

This paper examines the feasibility of extracting useful information from customer comments using a Naïve Bayes classifier. This was done for a database, obtained from a large Korean mobile telephone service provider, of 533 customer calls to call centers in 2009. After eliminating calls not containing customer complaints or comments, the remaining 383 comments were classified by an expert panel into four domains and 27 complaint categories. The four domains were Transaction‐related (189 comments, 49%), Product‐related (120 comments, 31%), Customer Service or Support‐related (38 comments, 10%) and Customer Outreach and Marketing‐related (36 comments, 9%). The comments were then randomly assigned to either a training set (257 cases, 67%) or test set (126 cases, 33%). The training set was used to develop a Naïve Bayes classifier that correctly predicted the domain 75% of the time and the specific subcategory 51% of the time for the test set. Prediction accuracy was strongly related to prediction strength for both sets of predictions, suggesting that simple filtering strategies where difficult to understand comments are flagged for expert review and easy comments are automatically classified are both technically feasible and likely to be practically valuable. Several strong predictors were also identified that corresponded to categories more detailed than those originally assigned. © 2012 Wiley Periodicals, Inc.     

Effect of Bacterial Infection on Ghrelin Receptor Regulation in Periodontal Cells and Tissues

The effect of bacterial infection on the expression of growth hormone secretagogue receptor (GHS-R) was investigated in periodontal cells and tissues, and the actions of ghrelin were evaluated. GHS-R was assessed in periodontal tissues of rats with and without periodontitis. Human gingival fibroblasts (HGFs) were exposed to Fusobacterium nucleatum in the presence and absence of ghrelin. GHS-R expression was determined by real-time PCR and immunocytochemistry. Furthermore, wound healing, cell viability, proliferation, and migration were evaluated. GHS-R expression was significantly higher at periodontitis sites as compared to healthy sites in rat tissues. F. nucleatum significantly increased the GHS-R expression and protein level in HGFs. Moreover, ghrelin significantly abrogated the stimulatory effects of F. nucleatum on CCL2 and IL-6 expressions in HGFs and did not affect cell viability and proliferation significantly. Ghrelin stimulated while F. nucleatum decreased wound closure, probably due to reduced cell migration. Our results show original evidence that bacterial infection upregulates GHS-R in rat periodontal tissues and HGFs. Moreover, our study shows that ghrelin inhibited the proinflammatory actions of F. nucleatum on HGFs without interfering with cell viability and proliferation, suggesting that ghrelin and its receptor may act as a protective molecule during bacterial infection on periodontal cells.     

Enzymatic membrane reactors: the determining factors in two separate phase operations

A two separate phase‐enzymatic membrane reactor is an attractive process since it has a large interfacial area and exchange surfaces, simultaneous reaction and separation and other benefits. Many factors influence its successful operation, and these include characteristics of the enzyme, membrane, circulating fluids and reactor operations. Although the operating conditions are the main factor, other factors must be considered before, during or after its application. At the initial stage of reactor development, the solubility of substrates and products, type of operation, membrane material and size, enzyme preparation and loading procedure, and cleanliness of the recirculated fluids should be specified. The immobilization site, reactor arrangement, dissolved or no‐solvent operation, classic or emulsion operation and immobilized or suspended enzyme(s) are determined later. Some factors still need further studies. Utilization of the technology is described for use from multigram‐ to plant‐scale capacity to process racemic and achiral compounds. The racemates were resolved primarily by kinetic resolution, but dynamic kinetic resolution has been exploited. The technology focused on hydrolytic reactions, but esterification processes were also exploited. Copyright © 2011 Society of Chemical Industry     

Thermal wave imaging through radio frequency induction heating

Radio frequency (RF) induction heating is used in order to generate thermal waves in conducting materials. This technique is applied to measure the thickness of tungsten-carbide plasma coating on stainless-steel plates. The letter briefly discusses major advantages and disadvantages of RF induction heating.     

Understanding Our Students: A Longitudinal‐Study of Success and Failure in Engineering With Implications for Increased Retention

In spite of considerable research about the poor retention rate of undergraduate engineering students, we still have an inadequate understanding of the factors that affect students' decisions to remain in engineering programs and their ability to perform well enough to be retained. Although continued study is needed of external factors such as curricular requirements, admissions criteria, and test scores, we also need to know much more about the relationships between curricular experiences and students' learning styles, habits, and attitudes. The work presented in this paper was designed to enhance educators' understanding of the factors that underlie the concern about student retention in engineering. By observing 1,000 engineering students during their first three years in college, the research team generated a large database on the students' academic and non‐academic characteristics as well as their successes and failures. The traits discovered not only support many findings from previous studies but also reveal some new relationships that could prove essential to designing an educational environment that will prepare engineers for success in the future.