Negligible depletion solid-phase microextraction with radiolabeled analytes to study free concentrations and protein binding: an example with [3H]estradiol

A new method is presented that enables sensitive measurement of free concentrations of radiolabeled ligands. Additionally, protein binding of radiochemicals in complex matrixes can be determined with this new technique that combines negligible depletion solid-phase microextraction (nd-SPME) with liquid scintillation counting (LSC) as detection. [3H]Estradiol was taken as an example compound. Possible matrix effects of protein on fiber uptake kinetics were studied. No matrix effect was found, either by fouling of the fiber, or by changed uptake kinetics. The validity of the method was shown in the determination of the affinity constant (Ka) of estradiol for human serum albumin (HSA). The Ka was estimated at 8.9 x 10(4) M(-1), which corresponds well with literature values. This study shows that nd-SPME is suitable to study the free concentration and protein binding of [3H]estradiol. The method described in this paper combines the advantages of nd-SPME with the advantages of radiolabeled analytes, creating a timesaving, simple, and sensitive analytical tool that will be particularly useful in complex matrixes containing many potential interferences for chromatographic methods.     

Dynamical diseases of brain systems: different routes to epileptic seizures

In this overview, we consider epilepsies as dynamical diseases of brain systems since they are manifestations of the property of neuronal networks to display multistable dynamics. To illustrate this concept we may assume that at least two states of the epileptic brain are possible: the interictal state characterized by a normal, apparently random, steady-state electroencephalography (EEG) ongoing activity, and the ictal state, that is characterized by paroxysmal occurrence of synchronous oscillations and is generally called, in neurology, a seizure. The transition between these two states can either occur: 1) as a continuous sequence of phases, like in some cases of mesial temporal lobe epilepsy (MTLE); or 2) as a sudden leap, like in most cases of absence seizures. In the mathematical terminology of nonlinear systems, we can say that in the first case the system's attractor gradually deforms from an interictal to an ictal attractor. The causes for such a deformation can be either endogenous or external. In this type of ictal transition, the seizure possibly may be anticipated in its early, preclinical phases. In the second case, where a sharp critical transition takes place, we can assume that the system has at least two simultaneous interictal and ictal attractors all the time. To which attractor the trajectories converge, depends on the initial conditions and the system's parameters. An essential question in this scenario is how the transition between the normal ongoing and the seizure activity takes place. Such a transition can occur either due to the influence of external or endogenous factors or due to a random perturbation and, thus, it will be unpredictable. These dynamical changes may not be detectable from the analysis of the ongoing EEG, but they may be observable only by measuring the system's response to externally administered stimuli. In the special cases of reflex epilepsy, the leap between the normal ongoing attractor and the ictal attractor is caused by a well-defined external perturbation. Examples from these different scenarios are presented and discussed.     

Ultra-Low-Power Interface Chip for Autonomous Capacitive Sensor Systems

Traditionally, most of the sensor interfaces must be tailored towards a specific application. This approach results in a high recurrent design cost and time to market. On the other hand, generic sensor interface design reduces the costs and offers a handy solution for multisensor applications. This paper presents a generic sensor interface chip (GSIC), which can read out a broad range of capacitive sensors. It contains capacitance-to-voltage converters, a switched-capacitor amplifier, an analog-to-digital converter, oscillators, clock generation circuits and a reference circuit. The system combines a very low-power design with a smart energy management, which adapts the current consumption according to the accuracy and speed requirements of the application. The GSIC is used in a pressure and an acceleration monitoring system. The pressure monitoring system achieves a current drain of 2.3 muA for a 10-Hz sample frequency and an 8-bit accuracy. In the acceleration monitoring system, we measured a current of 3.3 muA for a sample frequency of 10 Hz and an accuracy of 9 bits     

Method for short-term prediction of milk yield at the quarter level to improve udder health monitoring

Udder health problems are often associated with milk losses. These losses are different between quarters, as infected quarters are affected both by systemic and pathogen-specific local effects, whereas noninfected quarters are only subject to systemic effects. To gain insight in these losses and the milk yield dynamics during disease, it is essential to have a reliable reference for quarter-level milk yield in an unperturbed state, mimicking its potential yield. We developed a novel methodology to predict this quarter milk yield per milking session, using an historical data set of 504 lactations collected on a test farm by an automated milking system from DeLaval (Tumba, Sweden). Using a linear mixed model framework in which covariates associated with the linearized Wood model and the milking interval are included, we were able to describe quarter-level yield per milking session with a proportional error below 10%. Applying this model enables us to predict the milk yield of individual quarters 1 to 50 d ahead with a mean prediction error ranging between 8 and 20%, depending on the amount of historical data available to estimate the random effect covariates for the predicted lactation. The developed methodology was illustrated using 2 examples for which quarter-level milk losses are calculated during clinical mastitis. These showed that the quarter-level mixed model allows us to gain insight in quarter lactation dynamics and enables to calculate milk losses in different situations.     

CrimeSPOT: A language and runtime for developing active wireless sensor network applications

Advances in wireless sensing and actuation technology allow embedding significant amounts of application logic inside wireless sensor networks. Such active WSN applications are more autonomous, but are significantly more complex to implement. Event-based middleware lends itself to implementing these applications. It offers developers fine-grained control over how an individual node interacts with the other nodes of the network. However, this control comes at the cost of event handlers which lack composability and violate software engineering principles such as separation of concerns. In this paper, we present CrimeSPOT as a domain-specific language for programming WSN applications on top of event-driven middleware. Its node-centric features enable programming a node’s interactions through declarative rules rather than event handlers. Its network-centric features support reusing code within and among WSN applications. Unique to CrimeSPOT is its support for associating application-specific semantics with events that carry sensor readings. These preclude transposing existing approaches that address the shortcomings of event-based middleware to the domain of wireless sensor networks. We provide a comprehensive overview of the language and the implementation of its accompanying runtime. The latter comprises several extensions to the Rete forward chaining algorithm. We evaluate the expressiveness of the language and the overhead of its runtime using small, but representative active WSN applications.     

Microblog language identification: overcoming the limitations of short, unedited and idiomatic text

Multilingual posts can potentially affect the outcomes of content analysis on microblog platforms. To this end, language identification can provide a monolingual set of content for analysis. We find the unedited and idiomatic language of microblogs to be challenging for state-of-the-art language identification methods. To account for this, we identify five microblog characteristics that can help in language identification: the language profile of the blogger (blogger), the content of an attached hyperlink (link), the language profile of other users mentioned (mention) in the post, the language profile of a tag (tag), and the language of the original post (conversation), if the post we examine is a reply. Further, we present methods that combine these priors in a post-dependent and post-independent way. We present test results on 1,000 posts from five languages (Dutch, English, French, German, and Spanish), which show that our priors improve accuracy by 5 % over a domain specific baseline, and show that post-dependent combination of the priors achieves the best performance. When suitable training data does not exist, our methods still outperform a domain unspecific baseline. We conclude with an examination of the language distribution of a million tweets, along with temporal analysis, the usage of twitter features across languages, and a correlation study between classifications made and geo-location and language metadata fields.     

Understanding TES microcalorimeter noise and energy resolution

We have performed numerical calculations of the noise in voltage-biased superconducting transition edge-based X-ray microcalorimeters, using a finite-element model. Details of the model are discussed, as well as results for different absorber geometries. The results are in agreement with an analytical model and show that the amount of internal thermal fluctuation noise can be reduced by using a segmented absorber. The simulation also agrees well with experimental data, which, for our detectors, contain no major unidentified noise sources. Furthermore, the discrepancy between the (small-signal) theoretical and the measured energy resolutions for 5.9 keV X-rays, for our sensors typically a factor of 2, is explained by a more accurate modelling of the sensor responsivity, taking into account the large signal behaviour.     

Secreted production of a custom-designed, highly hydrophilic gelatin in Pichia pastoris

A custom-designed, highly hydrophilic gelatin was produced inPichia pastoris. Secreted production levels in single-copytransformants were in the range 3–6 g/l of clarified brothand purification to near homogeneity could be accomplished bydifferential ammonium sulfate precipitation. Despite the factthat gelatins are highly susceptible to proteolysis becauseof their unfolded structure, the recombinant protein was shownto be fully intact by SDS–PAGE, N-terminal sequencing,gel filtration chromatography and mass spectrometry. Owing toits highly hydrophilic nature, the migration of the syntheticgelatin in SDS–PAGE was severely delayed. Esterificationof the carboxylic amino acid side chains resulted in normalmigration. The high polarity of the synthetic gelatin also accountsfor its negligible surface activity in water at concentrationsup to 5% (w/v), as determined by tensiometry. Circular dichroismspectrometry showed that the non-hydroxylated gelatin did notform triple helices at 4°C. The spectrum was even more representativeof the random coil conformation than the spectrum of naturalnon-hydroxylated gelatins.