S100A6 Amyloid Fibril Formation Is Calcium-modulated and Enhances Superoxide Dismutase-1 (SOD1) Aggregation

S100A6 is a small EF-hand calcium- and zinc-binding protein involved in the regulation of cell proliferation and cytoskeletal dynamics. It is overexpressed in neurodegenerative disorders and a proposed marker for Amyotrophic Lateral Sclerosis (ALS). Following recent reports of amyloid formation by S100 proteins, we investigated the aggregation properties of S100A6. Computational analysis using aggregation predictors Waltz and Zyggregator revealed increased propensity within S100A6 helices HI and HIV. Subsequent analysis of Thioflavin-T binding kinetics under acidic conditions elicited a very fast process with no lag phase and extensive formation of aggregates and stacked fibrils as observed by electron microscopy. Ca2+ exerted an inhibitory effect on the aggregation kinetics, which could be reverted upon chelation. An FT-IR investigation of the early conformational changes occurring under these conditions showed that Ca2+ promotes anti-parallel β-sheet conformations that repress fibrillation. At pH 7, Ca2+ rendered the fibril formation kinetics slower: time-resolved imaging showed that fibril formation is highly suppressed, with aggregates forming instead. In the absence of metals an extensive network of fibrils is formed. S100A6 oligomers, but not fibrils, were found to be cytotoxic, decreasing cell viability by up to 40%. This effect was not observed when the aggregates were formed in the presence of Ca2+. Interestingly, native S1006 seeds SOD1 aggregation, shortening its nucleation process. This suggests a cross-talk between these two proteins involved in ALS. Overall, these results put forward novel roles for S100 proteins, whose metal-modulated aggregation propensity may be a key aspect in their physiology and function.

Specific label-free and real-time detection of oxidized low density lipoprotein (oxLDL) using an immunosensor with three monoclonal antibodies

Increased levels of plasma oxLDL, which is the oxidized fraction of Low Density Lipoprotein (LDL), are associated with atherosclerosis, an inflammatory disease, and the subsequent development of severe cardiovascular diseases that are today a major cause of death in modern countries. It is therefore important to find a reliable and fast assay to determine oxLDL in serum. A new immunosensor employing three monoclonal antibodies (mAbs) against oxLDL is proposed in this work as a quick and effective way to monitor oxLDL. The oxLDL was first employed to produce anti-oxLDL monoclonal antibodies by hybridoma cells that were previously obtained. The immunosensor was set-up by selfassembling cysteamine (Cyst) on a gold (Au) layer (4 mm diameter) of a disposable screen-printed electrode. Three mAbs were allowed to react with N-hydroxysuccinimide (NHS) and ethyl(dimethylaminopropyl)carbodiimide (EDAC), and subsequently incubated in the Au/Cys. Albumin from bovine serum (BSA) was immobilized further to ensure that other molecules apart from oxLDL could not bind to the electrode surface. All steps were followed by various characterization techniques such as electrochemical impedance spectroscopy (EIS) and square wave voltammetry (SWV). The analytical operation of the immunosensor was obtained by incubating the sensing layer of the device in oxLDL for 15 minutes, prior to EIS and SWV. This was done by using standard oxLDL solutions prepared in foetal calf serum, in order to simulate patient's plasma with circulating oxLDL. A sensitive response was observed from 0.5 to 18.0 mg mL 1 . The device was successfully applied to determine the oxLDL fraction in real serum, without prior dilution or necessary chemical treatment. The use of multiple monoclonal antibodies on a biosensing platform seemed to be a successful approach to produce a specific response towards a complex multi-analyte target, correlating well with the level of oxLDL within atherosclerosis disease, in a simple, fast and cheap way.

Virtual power players dealing with excessive wind power situations using real time pricing

The increasing importance given by environmental policies to the dissemination and use of wind power has led to its fast and large integration in power systems. In most cases, this integration has been done in an intensive way, causing several impacts and challenges in current and future power systems operation and planning. One of these challenges is dealing with the system conditions in which the available wind power is higher than the system demand. This is one of the possible applications of demand response, which is a very promising resource in the context of competitive environments that integrates even more amounts of distributed energy resources, as well as new players. The methodology proposed aims the maximization of the social welfare in a smart grid operated by a virtual power player that manages the available energy resources. When facing excessive wind power generation availability, real time pricing is applied in order to induce the increase of consumption so that wind curtailment is minimized. The proposed method is especially useful when actual and day-ahead wind forecast differ significantly. The proposed method has been computationally implemented in GAMS optimization tool and its application is illustrated in this paper using a real 937-bus distribution network with 20310 consumers and 548 distributed generators, some of them with must take contracts.

Ethernet goes real-time: a survey on research and technological developments

Ethernet is the most popular LAN technology. Its low price and robustness, resulting from its wide acceptance and deployment, has created an eagerness to expand its responsibilities to the factory-floor, where real-time requirements are to be fulfilled. However, it is difficult to build a real-time control network using Ethernet, because its MAC protocol, the 1-persistent CSMA/CD protocol with the BEB collision resolution algorithm, has unpredictable delay characteristics. Many anticipate that the recent technological advances in Ethernet such as the emerging Fast/Gigabit Ethernet, micro-segmentation and full-duplex operation using switches will also enable it to support time-critical applications. This technical report provides a comprehensive look at the unpredictability inherent to Ethernet and at recent technological advances towards real-time operation.

Assigning real-time tasks on heterogeneous multiprocessors with two unrelated types of processors

A preliminary version of this paper appeared in Proceedings of the 31st IEEE Real-Time Systems Symposium, 2010, pp. 239–248.

Real-time scheduling with resource sharing on uniform multiprocessors

Consider the problem of scheduling a set of implicit-deadline sporadic tasks to meet all deadlines on a uniform multiprocessor platform where each task may access at most one of |R| shared resources and at most once by each job of that task. The resources have to be accessed in a mutually exclusive manner. We propose an algorithm, GIS-vpr, which offers the guarantee that if a task set is schedulable to meet deadlines by an optimal task assignment scheme that allows a task to migrate only when it accesses or releases a resource, then our algorithm also meets the deadlines with the same restriction on the task migration, if given processors 4 + 6|R| times as fast. The proposed algorithm, by design, limits the number of migrations per job to at most two. To the best of our knowledge, this is the first result for resource sharing on uniform multiprocessors with proven performance guarantee.

Assigning real-time tasks on heterogeneous multiprocessors with two types of processors

Consider the problem of scheduling a set of implicitdeadline sporadic tasks on a heterogeneous multiprocessor so as to meet all deadlines. Tasks cannot migrate and the platform is restricted in that each processor is either of type-1 or type-2 (with each task characterized by a different speed of execution upon each type of processor). We present an algorithm for this problem with a timecomplexity of O(n·m), where n is the number of tasks and m is the number of processors. It offers the guarantee that if a task set can be scheduled by any non-migrative algorithm to meet deadlines then our algorithm meets deadlines as well if given processors twice as fast. Although this result is proven for only a restricted heterogeneous multiprocessor, we consider it significant for being the first realtime scheduling algorithm to use a low-complexity binpacking approach to schedule tasks on a heterogeneous multiprocessor with provably good performance.

Flexible and dynamic replication control for interdependent distributed real-time embedded systems

Replication is a proven concept for increasing the availability of distributed systems. However, actively replicating every software component in distributed embedded systems may not be a feasible approach. Not only the available resources are often limited, but also the imposed overhead could significantly degrade the system’s performance. This paper proposes heuristics to dynamically determine which components to replicate based on their significance to the system as a whole, its consequent number of passive replicas, and where to place those replicas in the network. The activation of passive replicas is coordinated through a fast convergence protocol that reduces the complexity of the needed interactions among nodes until a new collective global service solution is determined.

Robotic manipulators with vibrations: short time fourier transform of fractional spectra

This paper analyzes the signals captured during impacts and vibrations of a mechanical manipulator. In order to acquire and study the signals an experimental setup is implemented. The signals are treated through signal processing tools such as the fast Fourier transform and the short time Fourier transform. The results show that the Fourier spectrum of several signals presents a non integer behavior. The experimental study provides valuable results that can assist in the design of a control system to deal with the unwanted effects of vibrations.

Run-time defragmentation for dynamically reconfigurable hardware

Reconfigurable computing experienced a considerable expansion in the last few years, due in part to the fast run-time partial reconfiguration features offered by recent SRAM-based Field Programmable Gate Arrays (FPGAs), which allowed the implementation in real-time of dynamic resource allocation strategies, with multiple independent functions from different applications sharing the same logic resources in the space and temporal domains. However, when the sequence of reconfigurations to be performed is not predictable, the efficient management of the logic space available becomes the greatest challenge posed to these systems. Resource allocation decisions have to be made concurrently with system operation, taking into account function priorities and optimizing the space currently available. As a consequence of the unpredictability of this allocation procedure, the logic space becomes fragmented, with many small areas of free resources failing to satisfy most requests and so remaining unused. A rearrangement of the currently running functions is therefore necessary, so as to obtain enough contiguous space to implement incoming functions, avoiding the spreading of their components and the resulting degradation of system performance. A novel active relocation procedure for Configurable Logic Blocks (CLBs) is herein presented, able to carry out online rearrangements, defragmenting the available FPGA resources without disturbing functions currently running.

Fast mobility support in low-power wireless networks: smart-HOP over RPL/6LoWPAN

With the emergence of low-power wireless hardware new ways of communication were needed. In order to standardize the communication between these low powered devices the Internet Engineering Task Force (IETF) released the 6LoWPAN stand- ard that acts as an additional layer for making the IPv6 link layer suitable for the lower-power and lossy networks. In the same way, IPv6 Routing Protocol for Low- Power and Lossy Networks (RPL) has been proposed by the IETF Routing Over Low power and Lossy networks (ROLL) Working Group as a standard routing protocol for IPv6 routing in low-power wireless sensor networks. The research performed in this thesis uses these technologies to implement a mobility process. Mobility management is a fundamental yet challenging area in low-power wireless networks. There are applications that require mobile nodes to exchange data with a xed infrastructure with quality-of-service guarantees. A prime example of these applications is the monitoring of patients in real-time. In these scenarios, broadcast- ing data to all access points (APs) within range may not be a valid option due to the energy consumption, data storage and complexity requirements. An alternative and e cient option is to allow mobile nodes to perform hand-o s. Hand-o mechanisms have been well studied in cellular and ad-hoc networks. However, low-power wireless networks pose a new set of challenges. On one hand, simpler radios and constrained resources ask for simpler hand-o schemes. On the other hand, the shorter coverage and higher variability of low-power links require a careful tuning of the hand-o parameters. In this work, we tackle the problem of integrating smart-HOP within a standard protocol, speci cally RPL. The simulation results in Cooja indicate that the pro- posed scheme minimizes the hand-o delay and the total network overhead. The standard RPL protocol is simply unable to provide a reliable mobility support sim- ilar to other COTS technologies. Instead, they support joining and leaving of nodes, with very low responsiveness in the existence of physical mobility.

Real-time scheduling with resource sharing on heterogeneous multiprocessors

Consider the problem of scheduling a task set τ of implicit-deadline sporadic tasks to meet all deadlines on a t-type heterogeneous multiprocessor platform where tasks may access multiple shared resources. The multiprocessor platform has m k processors of type-k, where k∈{1,2,…,t}. The execution time of a task depends on the type of processor on which it executes. The set of shared resources is denoted by R. For each task τ i , there is a resource set R i ⊆R such that for each job of τ i , during one phase of its execution, the job requests to hold the resource set R i exclusively with the interpretation that (i) the job makes a single request to hold all the resources in the resource set R i and (ii) at all times, when a job of τ i holds R i , no other job holds any resource in R i . Each job of task τ i may request the resource set R i at most once during its execution. A job is allowed to migrate when it requests a resource set and when it releases the resource set but a job is not allowed to migrate at other times. Our goal is to design a scheduling algorithm for this problem and prove its performance. We propose an algorithm, LP-EE-vpr, which offers the guarantee that if an implicit-deadline sporadic task set is schedulable on a t-type heterogeneous multiprocessor platform by an optimal scheduling algorithm that allows a job to migrate only when it requests or releases a resource set, then our algorithm also meets the deadlines with the same restriction on job migration, if given processors 4×(1+MAXP×⌈|P|×MAXPmin{m1,m2,…,mt}⌉) times as fast. (Here MAXP and |P| are computed based on the resource sets that tasks request.) For the special case that each task requests at most one resource, the bound of LP-EE-vpr collapses to 4×(1+⌈|R|min{m1,m2,…,mt}⌉). To the best of our knowledge, LP-EE-vpr is the first algorithm with proven performance guarantee for real-time scheduling of sporadic tasks with resource sharing on t-type heterogeneous multiprocessors.

Disposable immunosensor using a simple method for oriented antibody immobilization for label-free real-time detection of an oxidative stress biomarker implicated in cancer diseases

This work proposes a novel approach for a suitable orientation of antibodies (Ab) on an immunosensing platform, applied here to the determination of 8-hydroxy-2′-deoxyguanosine (8OHdG), a biomarker of oxidative stress that has been associated to chronic diseases, such as cancer. The anti-8OHdG was bound to an amine modified gold support through its Fc region after activation of its carboxylic functions. Non-oriented approaches of Ab binding to the platform were tested in parallel, in order to show that the presented methodology favored Ab/Ag affinity and immunodetection of the antigen. The immunosensor design was evaluated by quartz-crystal microbalance with dissipation, atomic force microscopy, electrochemical impedance spectroscopy (EIS) and square-wave voltammetry. EIS was also a suitable technique to follow the analytical behavior of the device against 8OHdG. The affinity binding between 8OHdG and the antibody immobilized in the gold modified platform increased the charge transfer resistance across the electrochemical set-up. The observed behavior was linear from 0.02 to 7.0 ng/mL of 8OHdG concentrations. The interference from glucose, urea and creatinine was found negligible. An attempt of application to synthetic samples was also successfully conducted. Overall, the presented approach enabled the production of suitably oriented Abs over a gold platform by means of a much simpler process than other oriented-Ab binding approaches described in the literature, as far as we know, and was successful in terms of analytical features and sample application.

Disposable immunosensor with simple antibody orientation for label-free real-time detection of a cancer biomarker

This work proposes a novel approach for a suitable orientation of antibodies (Ab) on an immunosensing platform, applied here to the determination of 8-hydroxy-2’-deoxyguanosine (8OHdG), a biomarker of oxidative stress that has been associated to chronic diseases, such as cancer. The Anti-8OHdG was bound to an amine modified gold support through its Fc region after activation of its carboxylic functions. Non-oriented approaches of Ab binding to the platform were tested in parallel, in order to show that the presented proposal favored Ab/Ag affinity. The immunosensor design was evaluated by Quartz-Crystal microbalance with Dissipation, Atomic Force Microscopy, Electrochemical Impedance Spectroscopy (EIS) and Square-Wave Voltammetry. EIS was also a suitable technique to follow the analytical behavior of the device against 8OHdG. The affinity binding between 8OHdG and the antibody immobilized in the gold modified platform increased the charged transfer resistance across the electrochemical sep-up. The observed behavior was linear from 0.02 to 7.0 ng/mL of 8OHdG concentrations. The interference from Glucose, Urea and Creatinine was found negligible. An attempt of application to synthetic samples was also successfully conducted. Overall, the presented approach enabled the production of suitably oriented Abs over a gold platform by means of a much simpler process than other oriented-Ab binding approaches described in the literature, as far as we know, and was successful in terms of analytical features and sample application.