NMNAT1 Is a Survival Factor in Actinomycin D-Induced Osteosarcoma Cell Death

Osteosarcoma is a frequent and extremely aggressive type of pediatric cancer. New therapeutic approaches are needed to improve the overall survival of osteosarcoma patients. Our previous results suggest that NMNAT1, a key enzyme in nuclear NAD⁺ synthesis, facilitates the survival of cisplatin-treated osteosarcoma cells. A high-throughput cytotoxicity screening was performed to identify novel pathways or compounds linked to the cancer-promoting role of NMNAT1. Nine compounds caused higher toxicity in the NMNAT1 KO U2OS cells compared to their wild type counterparts, and actinomycin D (ActD) was the most potent. ActD-treatment of NMNAT1 KO cells increased caspase activity and secondary necrosis. The reduced NAD⁺ content in NMNAT1 KO cells was further decreased by ActD, which partially inhibited NAD⁺-dependent enzymes, including the DNA nick sensor enzyme PARP1 and the NAD⁺-dependent deacetylase SIRT1. Impaired PARP1 activity increased DNA damage in ActD-treated NMNAT1 knockout cells, while SIRT1 impairment increased acetylation of the p53 protein, causing the upregulation of pro-apoptotic proteins (NOXA, BAX). Proliferation was decreased through both PARP- and SIRT-dependent pathways. On the one hand, PARP inhibitors sensitized wild type but not NMNAT1 KO cells to ActD-induced anti-clonogenic effects; on the other hand, over-acetylated p53 induced the expression of the anti-proliferative p21 protein leading to cell cycle arrest. Based on our results, NMNAT1 acts as a survival factor in ActD-treated osteosarcoma cells. By inhibiting both PARP1- and SIRT1-dependent cellular pathways, NMNAT1 inhibition can be a promising new tool in osteosarcoma chemotherapy.     

Effect of Inducible BMP-7 Expression on the Osteogenic Differentiation of Human Dental Pulp Stem Cells

BMP-7 has shown inductive potential for in vitro osteogenic differentiation of mesenchymal stem cells, which are an ideal resource for regenerative medicine. Externally applied, recombinant BMP-7 was able to induce the osteogenic differentiation of DPSCs but based on our previous results with BMP-2, we aimed to study the effect of the tetracyclin-inducible BMP-7 expression on these cells. DPSC, mock, and DPSC-BMP-7 cell lines were cultured in the presence or absence of doxycycline, then alkaline phosphatase (ALP) activity, mineralization, and mRNA levels of different osteogenic marker genes were measured. In the DPSC-BMP-7 cell line, the level of BMP-7 mRNA significantly increased in the media supplemented with doxycycline, however, the expression of Runx2 and noggin genes was upregulated only after 21 days of incubation in the osteogenic medium with doxycycline. Moreover, while the examination of ALP activity showed reduced activity in the control medium containing doxycycline, the accumulation of minerals remained unchanged in the cultures. We have found that the induced BMP-7 expression failed to induce osteogenic differentiation of DPSCs. We propose three different mechanisms that may worth investigating for the engineering of expression systems that can be used for the induction of differentiation of mesenchymal stem cells.     

Application of probabilistic and fuzzy cognitive approaches in semantic web framework for medical decision support

This study aimed to focus on medical knowledge representation and reasoning using the probabilistic and fuzzy influence processes, implemented in the semantic web, for decision support tasks. Bayesian belief networks (BBNs) and fuzzy cognitive maps (FCMs), as dynamic influence graphs, were applied to handle the task of medical knowledge formalization for decision support. In order to perform reasoning on these knowledge models, a general purpose reasoning engine, EYE, with the necessary plug-ins was developed in the semantic web. The two formal approaches constitute the proposed decision support system (DSS) aiming to recognize the appropriate guidelines of a medical problem, and to propose easily understandable course of actions to guide the practitioners. The urinary tract infection (UTI) problem was selected as the proof-of-concept example to examine the proposed formalization techniques implemented in the semantic web. The medical guidelines for UTI treatment were formalized into BBN and FCM knowledge models. To assess the formal models’ performance, 55 patient cases were extracted from a database and analyzed. The results showed that the suggested approaches formalized medical knowledge efficiently in the semantic web, and gave a front-end decision on antibiotics’ suggestion for UTI.     

Initial column profile with composition-dependent relative volatility

When solving the large scale, highly nonlinear, equation systems of steady state multistage distillation processes modelled with theoretical plates, convergence depends on the initial values. The conventionally applied linear initial profiles frequently give rise to divergence when azeotrope is present. The usual practice in such cases is initializing with engineering insight, i.e. anticipating the results. Composition-dependent relative volatility model is developed in the present work, and applied successfully to initialize the composition profiles of distillation columns. The model describes the implicit temperature dependence through composition dependence. Equlibrium plots computed with the new model well approximate the measured data of strongly non-ideal, and even azeotropic, mixtures. The initial column profiles computed with the new model are rather similar to the final solution, and thus accelerate the computation. Processes can be modelled with the new initial profile when the solution algorithms do not converge with the conventional initialization.     

Electrosynthesis and comparative studies on carboxyl-functionalized polythiophene derivatives

Electrochemical synthesis of a novel carboxylic acid functionalized polythiophene – poly(3-thiophene-butyric-acid), PTBA – has been realized. Its morphology, electrochemical, spectral and conducting properties have been compared to those of poly(3-thiophene-acetic-acid), PTAA, which is widely used to immobilise both bioactive molecules and inorganic nanoparticles. According to scanning electron microscopic (SEM) images, the difference in the real and geometric surface area of the modified electrodes is much more expressed in the case of PTBA. Both the symmetry of the cyclic voltammograms and the concurrent, sustained optical changes proved that this polymer possesses an improved and more stable redox activity. According to simultaneously performed in situ ac. impedance and UV–Vis measurements, both films could be uniformly transformed between the insulating and conducting forms, but PTAA exhibited some degradation. The development of the conducting state during the redox switching of both thiophene derivative polymers proved to be primarily connected to the formation of di-cationic species. The electrochemical quartz crystal microbalance (EQCM) results evidenced also differences between the two polymers, which difference can be interpreted by assuming the more expressed effect of the deprotonation-connected (self-) doping process in PTAA. The results confirm that the new conducting polymer, PTBA is much more convenient for being considered as the polymer matrix of practically applicable composites.     

Micromagnetic simulation of current-driven domain wall propagation

We develop a method to model the interaction of currents and domain walls in a standard micromagnetic simulator. The interaction of spin-polarized currents and localized magnetic moments can be described by a current-induced effective field, which superposes to other components of the effective magnetic field. Using our method, we examine domain wall propagation in ideal, rough-surfaced and notched domain walls.     

2D operators on topographic and non‐topographic architectures—implementation,efficiency analysis,and architecture selection methodology

Topographic and non‐topographic image processing architectures and chips, developed within the CNN community recently, are analyzed and compared. It is achieved on a way that the 2D operators are collected to classes according to their implementation methods on the different architectures, and the main implementation parameters of the different operator classes are compared. Based on the results, an efficient architecture selection methodology is formalized. Copyright © 2010 John Wiley & Sons, Ltd.     

Field-coupled computing in magnetic multilayers

This paper presents a computational study of ion-beam patterned cobalt-platinum multilayers, which could be used for field-coupled computing. We use micromagnetic simulations to reproduce measured hysteresis curves. This parameterized micromagnetic simulator then will be used for simulating interacting magnetic dots. We demonstrate how logic gates can be built from such coupled dots. We also show how electrical wires—placed beneath or above the magnetic dots—can provide a magnetic field, which propagates the magnetic signals.