In silico and in vitro approaches to develop Dimethylarginine dimethylaminohydrolase-1 inhibitors

Introduction: Dimethylarginine dimethylaminohydrolases (DDAH) metabolise the endogenous nitric oxide synthase (NOS) inhibitors: asymmetric dimethylarginine (ADMA) and monomethylarginine1. In sepsis excessive nitric oxide partially contributes to acute circulatory failure, and pharmacological DDAH1 inhibition has been proposed in order to increase methylarginines and reduce NO levels 2. The SR257 arginine analogue, with NG-methoxyethyl substituent, inhibits DDAH1 with an IC50 22 µM without directly inhibiting NOSs1,3. Methods: Acyclic and cyclic NG,NG-disubstituted arginines were made as previously described4 using Katritzky’s synthesis preparing trisubstituted guanidines from di-(benzotriazol-1-yl)methanimine5. Molecular docking was employed to explore interactions of these NG,NG-disubstituted arginines with human DDAH1 (PDB 2JAJ) using Glide (Schroedinger6) and Autodock47. The published SR257 ligand was used to define the binding site with both software tools. Recombinant human DDAH1 activity was measured using colorometric citrulline assay8 containing ADMA (100 µM), sodium phosphate (10 mM pH7.4); with symmetric dimethylarginine (100 µM), not a substrate for DDAH1, as blank. Experiments were carried out in duplicate, and repeated on at least 3 separate occasions. Results: Recombinant DDAH1 activity was reduced to less than 25% of control (ADMA substrate, 100 µM) in the presence of 100 µM piperidinyl, methoxyethyl/methyl, N-methylpiperazinyl, with morpholinyl and pyrrolidinyl substituents reducing activity to less than 10% of control. The in silico Glide docking score and predicted Autodock4 binding energy for human DDAH1 (PDB, 2JAJ) for the known SR257 DDAH1 inhibitor and NG,NG-disubstituted arginines are shown in the table: Conclusion: Both Autodock4 and Glide docking predicted higher binding energies for morpholinyl, pyrrolidinyl and piperinyl than the known SR257 compound. In vitro assays confirmed these NG,NG-disubstituted arginines reduced DDAH1 activity. There was variation between Glide and Autodock4 in the docking predictions for methoxyethyl/methyl and N-methylpiperazinyl. In silico prediction of DDAH1-ligand interactions may assist in the future design and development of novel NG,NG-disubstituted arginines. References: 1 Leiper, J. et al. (2007) Nat Med. 13:198-203. 2 Wang, Z et al. (2014) Biochem J. 460:309 3 Rossiter, S. et al. (2005) J Med Chem. 48:4670-4678. 4 Morfill, C et al. (2012) http://www.pA2online.org/abstracts/Vol10Issue4abst197P.pdf 5 Katritzky, A et al. (2000) J. Org. Chem. 65: 8080-8082. 6 Friesner, RA et al. (2006) J Med Chem. 49:6177-6196. 7 Morris, GM et al. (2009) J. Comp. Chem. 16:2785-91. 8 Knipp, M & Vasak, M (2000) Anal Biochem 286:257-64

Fatigue detection using computer vision

Long duration driving is a significant cause of fatigue related accidents of cars, airplanes, trains and other means of transport. This paper presents a design of a detection system which can be used to detect fatigue in drivers. The system is based on computer vision with main focus on eye blink rate. We propose an algorithm for eye detection that is conducted through a process of extracting the face image from the video image followed by evaluating the eye region and then eventually detecting the iris of the eye using the binary image. The advantage of this system is that the algorithm works without any constraint of the background as the face is detected using a skin segmentation technique. The detection performance of this system was tested using video images which were recorded under laboratory conditions. The applicability of the system is discussed in light of fatigue detection for drivers

Hiding the squid:patterns in artificial cephalopod skin

Cephalopods employ their chromomorphic skins for rapid and versatile active camouflage and signalling effects. This is achieved using dense networks of pigmented, muscle-driven chromatophore cells which are neurally stimulated to actuate and affect local skin colouring. This allows cephalopods to adopt numerous dynamic and complex skin patterns, most commonly used to blend into the environment or to communicate with other animals. Our ultimate goal is to create an artificial skin that can mimic such pattern generation techniques, and that could produce a host of novel and compliant devices such as cloaking suits and dynamic illuminated clothing. This paper presents the design, mathematical modelling and analysis of a dynamic biomimetic pattern generation system using bioinspired artificial chromatophores. The artificial skin is made from electroactive dielectric elastomer: a soft, planar-actuating smart material that we show can be effective at mimicking the actuation of biological chromatophores. The proposed system achieves dynamic pattern generation by imposing simple local rules into the artificial chromatophore cells so that they can sense their surroundings in order to manipulate their actuation. By modelling sets of artificial chromatophores in linear arrays of cells, we explore the capability of the system to generate a variety of dynamic pattern types. We show that it is possible to mimic patterning seen in cephalopods, such as the passing cloud display, and other complex dynamic patterning

Temperature dependence of the resistance of metallic nanowires (diameter ≥\geq 15 nm): Applicability of Bloch-Gr\"{u}neisen theorem

We have measured the resistances (and resistivities) of Ag and Cu nanowires of diameters ranging from 15nm to 200nm in the temperature range 4.2K-300K with the specific aim to assess the applicability of the Bloch-Gr\"{u}neisen formula for electron phonon resistivity in these nanowires. The wires were grown within polymeric templates by electrodeposition. We find that in all the samples the resistance reaches a residual value at T=4.2K and the temperature dependence of resistance can be fitted to the Bloch-Gr\"{u}neisen formula in the entire temperature range with a well defined transport Debye temperature ($\Theta_{R}$). The value of Debye temperature obtained from the fits lie within 8% of the bulk value for Ag wires of diameter 15nm while for Cu nanowires of the same diameter the Debye temperature is significantly lesser than the bulk value. The electron-phonon coupling constants (measured by $\alpha_{el-ph}$ or $\alpha_{R}$) in the nanowires were found to have the same value as that of the bulk. The resistivities of the wires were seen to increase as the wire diameter was decreased. This increase in the resistivity of the wires may be attributed to surface scattering of conduction electrons. The specularity p was estimated to be about 0.5. The observed results allow us to obtain the resistivities exactly from the resistance and gives us a method of obtaining the exact numbers of wires within the measured array (grown within the template).Comment: 9 pages, 10 figure

Modelling and analysis of pH responsive hydrogels for the development of biomimetic photo-actuating structures

ABSTRACTPhoto-actuating structures inspired by the chemical sensing and signal transmission observed in sun-tracking leaves have recently been proposed by Dicker et al. The proposed light tracking structures are complex, multicomponent material systems, principally composed of a reversible photoacid or base, combined with a pH responsive hydrogel actuator. New modelling and characterization approaches for pH responsive hydrogels are presented in order to facilitate the development of the proposed structures. The model employs Donnan equilibrium for the prediction of hydrogel swelling in systems where the pH change is a variable resulting from the equilibrium interaction of all free and fixed (hydrogel) species. The model allows for the fast analysis of a variety of combinations of material parameters, allowing for the design space for the proposed photo-actuating structures to be quickly established. In addition, experimental examination of the swelling of a polyether-based polyurethane and poly(acrylic acid) interpenetrating network hydrogel is presented. The experiment involves simultaneously performing a titration of the hydrogel, and undertaking digital image correlation (DIC) to determine the hydrogel’s state of swelling. DIC allows for the recording of the hydrogel’s state of swelling with previously unattained levels of resolution. Experimental results provide both model material properties, and a means for model validation.</jats:p

Developing adaptive capability: the case of Nottingham’s nascent biotechnology sector

A region’s adaptive capability describes its ability to respond positively to changes and shocks that affect the regional economy over time and take advantage of new and emerging market opportunities from wherever they arise. Central to this idea is the manner in which historic economic strengths or capabilities can be turned to new purposes. This paper provides a case study that uses the concept of adaptive capability as a lens through which to explore the emergence of a notable bioscience based industrial cluster in a city region of the UK and the part that it played in helping to restructure the economic base of the city. The local economy of Nottingham faced major structural changes in the late 1990s. These changes saw not only the demise of what had at one time been a key industrial sector, but also the departure of two of the city’s three principal employers, and the closure of a major research facility owned by a third. The case explores the manner in which capabilities linked to this constellation of firms were redeployed in order to take advantage of new market opportunities. The case places particular emphasis on the contribution of firms, the restructuring of industry sectors and institutional changes that occurred at this time, to the city’s adaptive capability. In the process the case study reveals a notable example of adaptation as the local economy moved away from a previous path of regional and sectoral development, towards a new and yet related trajectory. Central to this process was the purposive re-tasking of physical assets and the mobilization of knowledge assets that were the legacy of one of the city’s historic industrial strengths. The paper concludes with a brief consideration of the wider relevance and applicability of this model of bioscience based regional development