Dual Knockdown of Musashi RNA-Binding Proteins MSI-1 and MSI-2 Attenuates Putative Cancer Stem Cell Characteristics and Therapy Resistance in Ovarian Cancer Cells

In ovarian cancer, therapy resistance mechanisms complicate cancer cell eradication. Targeting Musashi RNA-binding proteins (MSI) may increase therapeutic efficacy. Database analyses were performed to identify gene expression associations between MSI proteins and key therapy resistance and cancer stem cell (CSC) genes. Then, ovarian cancer cells were subjected to siRNA-based dual knockdown of MSI-1 and MSI-2. CSC and cell cycle gene expression was investigated using quantitative polymerase chain reaction (qPCR), western blots, and flow cytometry. Metabolic activity and chemoresistance were assessed by MTT assay. Clonogenic assays were used to quantify cell survival post-irradiation. Database analyses demonstrated positive associations between MSI proteins and putative CSC markers NOTCH, MYC, and ALDH4A1 and negative associations with NOTCH inhibitor NUMB. MSI-2 expression was negatively associated with the apoptosis regulator p21. MSI-1 and MSI-2 were positively correlated, informing subsequent dual knockdown experiments. After MSI silencing, CSC genes were downregulated, while cell cycle progression was reduced. Metabolic activity was decreased in some cancer cells. Both chemo- and radioresistance were reduced after dual knockdown, suggesting therapeutic potential. Dual knockdown of MSI proteins is a promising venue to impede tumor growth and sensitize ovarian cancer cells to irradiation and chemotherapy.     

Engineering model of a passive planar air breathing fuel cell cathode

The behavior of an air breathing fuel cell (ABFC) operated on dry-hydrogen in dead-ended mode is studied using theoretical analysis. A one-dimensional, non-isothermal, combined heat and mass transport model is developed that captures the coupling between water generation, oxygen consumption, self-heating and natural convection at the air breathing cathode. The model is validated against planar ABFC experimental measurements over a range of ambient temperatures. The model confirms the strong effect of self-heating on the water balance within passive ABFCs. Model analysis provides several conclusions: (1) thermal runaway caused by inadequate heat rejection predominantly limits ABFC performance. (2) The natural convection boundary layer represents a significant barrier to cathode mass and heat transfer. (3) Because the mass and heat transport numbers associated with natural convection are small, even slight forced convection dramatically affects cell behavior. (4) Performance optimization requires maximizing heat rejection while minimizing flooding. Decoupling the latter two phenomena is challenging due to the exponential relationship between water vapor saturation and temperature.     

Applications of one-cycle control to improve the interconnection of a solid oxide fuel cell and electric power system with a dynamic load

Adding distributed generation (DG) is a desirable strategy for providing highly efficient and environmentally benign services for electric power, heating, and cooling. The interface between a solid oxide fuel cell (SOFC), typical loads, and the electrical grid is simulated in Matlab/Simulink and analyzed to assess the interactions between DG and the electrical grid. A commercial building load profile is measured during both steady-state and transient conditions. The load data are combined with the following models that are designed to account for physical features: a One-Cycle Control grid-connected inverter, a One-Cycle Control active power filter, an SOFC, and capacitor storage. High penetration of DG without any power filter increases the percentage of undesirable harmonics provided by the grid, but combined use of an inverter and active power filter allows the DG system interconnection to improve the grid tie-line flow by lowering total harmonic distortion and increasing the power factor to unity.     

Reconstruction of graphs based on random walks

The analysis of complex networks is of major interest in various fields of science. In many applications we face the challenge that the exact topology of a network is unknown but we are instead given information about distances within this network. The theoretical approaches to this problem have so far been focusing on the reconstruction of graphs from shortest path distance matrices. Often, however, movements in networks do not follow shortest paths but occur in a random fashion. In these cases an appropriate distance measure can be defined as the mean length of a random walk between two nodes — a quantity known as the mean first hitting time.In this contribution we investigate whether a graph can be reconstructed from its mean first hitting time matrix and put forward an algorithm for solving this problem. A heuristic method to reduce the computational effort is described and analyzed. In the case of trees we can even give an algorithm for reconstructing graphs from incomplete random walk distance matrices.     

Diesel exhaust particles modulate the tight junction protein occludin in lung cells in vitro

Background  

Using an in vitro triple cell co-culture model consisting of human epithelial cells (16HBE14o-), monocyte-derived macrophages and dendritic cells, it was recently demonstrated that macrophages and dendritic cells create a transepithelial network between the epithelial cells to capture antigens without disrupting the epithelial tightness. The expression of the different tight junction proteins in macrophages and dendritic cells, and the formation of tight junction-like structures with epithelial cells has been demonstrated. Immunofluorescent methods combined with laser scanning microscopy and quantitative real-time polymerase chain reaction were used to investigate if exposure to diesel exhaust particles (DEP) (0.5, 5, 50, 125 μg/ml), for 24 h, can modulate the expression of the tight junction mRNA/protein of occludin, in all three cell types.     

Ex vivo and in vivo Evaluation of [18F]PR04.MZ in Rodents: A Selective Dopamine Transporter Imaging Agent

N‐4‐Fluorobut‐2‐yn‐1‐yl‐2β‐carbomethoxy‐3β‐phenyltropane (PR04.MZ) has been developed as dopamine transporter (DAT) ligand for molecular imaging. It contains a terminally fluorinated, conformationally constrained nitrogen substituent that is well suited for the introduction of fluorine‐18. The present report describes the pharmacological characterisation of [¹⁸F]PR04.MZ. The ligand shows an IC₅₀ value of 2 nM against human DAT, whereas the IC₅₀ value against human serotonin transporter and human noradrenalin transporter are lower (110 nM and 22 nM , respectively). Furthermore, its ex vivo organ distribution, its binding profile in the rat brain and reversibility of binding were examined. A μPET study illuminates a fast kinetic profile and specific binding to rat DAT.     

Making Epothilones Fluoresce: Design,Synthesis, and Biological Characterization of a Fluorescent N12‐Aza‐Epothilone (Azathilone)

A green fluorescent 12‐aza‐epothilone (azathilone) derivative has been prepared through the attachment of the 4‐nitro‐2,1,3‐benzoxadiazole (NBD) fluorophore to the 12‐nitrogen atom of the azamacrolide core structure. While less potent than natural epothilones or different N12‐acylated azathilone derivatives, NBD‐azathilone ( 3 ) promotes tubulin assembly, inhibits cancer cell proliferation in vitro and arrests the cell cycle at the G2/M transition. Most significantly, the binding of 3 to cellular microtubules (MTs) could be directly visualized by confocal fluorescence microscopy. Based on competition binding experiments with laulimalide‐stabilized MTs in vitro, the N12‐Boc substituted azathilone 1 , Epo A, and NBD‐azathilone ( 3 ) all interact with the same tubulin‐binding site. Computational studies provided a structural model of the complexes between β‐tubulin and 1 or 3 , respectively, in which the NBD moiety of 3 or the BOC moiety of 1 directly and specifically contribute to MT binding. Collectively, these data demonstrate that the cellular effects of 3 and, by inference, also of other azathilones are the result of their interactions with the cellular MT network.     

Transforming the cell surface through proteolysis

Protein shedding, or the proteolytic cleavage of a protein from the surface of a cell, is emerging as an important mechanism in the regulation of cellular activity but it is poorly understood. Growing evidence suggesting that protein shedding and protein function are closely linked may lead to new strategies for the treatment of a wide range of diseases.     

Biomechanical assessment of new hand-powered pruning shears

The aim of the study was to assess the biomechanical strains on the hand-wrist system during grapevine pruning with newly designed pruning shears. Surface electromyography of the finger flexor muscle and wrist postures were analyzed in four vineyard workers during actual work with the new and reference hand-powered pruning shears. Due to the large inter-individual differences in biomechanical data observed with both pruning shears, no systematic effect of the new pruning shears on "extreme" flexion/extension (F/E) and ulnar/radial (U/R) wrist deviations was demonstrated. However, the use of the new pruning shears was associated with a higher frequency of "neutral and moderate" U/R deviations (U/R < 20%(max)) and, to a lesser extent, of "neutral and moderate" F/E deviations (F/E < 20%(max)). Due to the small sample of workers, this result must be confirmed in a larger population of vineyard workers.