Dithiobissuccinimidyl propionate as an anchor for assembling peroxidases at electrodes surfaces and its application in a H2O2 biosensor

Exposure of gold surfaces to solutions of dithiobis N-succinimidyl propionate (DTSP) gives rise to the modification of the surface with N-succinimidyl-3-thiopropionate (NSTP) which can, in turn, react with amino groups allowing for the covalent immobilization of enzymes such as horseradish peroxidase (HRP). The coverage of NSTP has been estimated to be of the order of 1.3 x 10(-10) from the charge consumed during its reductive desorption. The binding reaction of HRP with NSTP modified gold surfaces has been studied with the quartz crystal microbalance, and the results suggest that the immobilization process involves two steps in which the first (faster) appears to correspond to the rapid incorporation of the enzyme whereas the second is likely due to the slow incorporation of additional enzyme and/or reorganization of the immobilized layer. Spectrophotometric and electrochemical assays indicate that the immobilized HRP retains its enzymatic activity after immobilization onto the DTSP modified gold surface. The amount of immobilized (and active) HRP was estimated from QCM and spectrophotometric measurements to be of the order of 1.5 x 10(-11) mol/cm2. A peroxide biosensor was developed making use of a gold surface modified with DTSP and HRP employing Os and Ru complexes of 1,10-phenanthroline 5,6-dione (phen-dione) of the type [M(phendione)x(L)3-x]+2 (where L = 1,10-phenanthroline or 2,2'-bipyridine, x = 1-3) as mediators with the quinone moieties being the active component. The efficiency of the mediators increased with increasing number of phendione ligands.     

Septo-optic dysplasia: report of 6 patients studied with MR and discussion on its pathogenesis

OBJECTIVE: Septo-optic dysplasia, which consists of the association of the hypoplasia of the optic nerves and the agenesis of the septum pellucidum, is frequently associated with deficiency of hypothalamic releasing factors. In Magnetic Resonance (MR) of these patients, anomalies in the form and size of the pituitary stalk, adenohypophysis and neurohypophysis are found. Some cases show schizencephaly and it has been proposed as an added component of the syndrome by some authors. This fact has been refuted by others. PATIENTS AND METHODS: We present the clinical and neuroanatomic revision of six children with septo-optic dysplasia studied by MR imaging over the last five years in our Department of Neuropediatrics. The aim was, that through the neuroembryological discussion of the morphopathological aspects of the patients, to determine the malformation and the time in which the injury, which was the underlying cause, occurred. RESULTS: From the six cases, in two only disruptive signs were evident with the optic nerves being affected asymmetrically, disruption of the corpus callosum, falx cerebri indemnity and effects in the cortex conformation. This was opposed to the dysgenic features in the other four cases which had no disruptive features. CONCLUSIONS: Our findings suggest that this entity could be the result of at least two different pathogenic processes, that is, a minor form of holoprosencephaly (dysgenesis) or a disruption which, therefore, occurs later in gestation.     

A novel cartilage protein (CILP) present in the mid-zone of human articular cartilage increases with age

A novel, somewhat basic noncollagenous protein was purified from guanidine hydrochloride extracts of human articular cartilage using cesium chloride density gradient centrifugation, followed by ion-exchange chromatography at pH 5, and gel filtration on two serially coupled columns of Superose 6 and Superdex 200. The protein of 91.5 kDa contains a single polypeptide chain substituted with N-linked oligosaccharides. It appeared unique to cartilage as studied by enzyme-linked immunosorbent assay and immunoblots of various tissue extracts. Its concentration in articular cartilages showed some variability with age being lower in young individuals. It represents a chondrocyte product, since it is synthesized by articular chondrocytes in explant cultures. Interestingly, the distribution of the protein in the articular cartilage provides important information on the nature of chondrocytes at different compartments in the tissue. Thus, chondrocytes in the middle/deeper layers of the tissue in particular, appeared to have produced the protein and deposited it in the interterritorial matrix. The protein was neither seen in the superficial nor in the deepest regions of the articular cartilage. Based on its immunolocalization we have named this protein CILP (cartilage intermediate layer protein).     

Cellular localization and changes in expression of prolactin receptor isoforms in sheep ovary throughout the estrous cycle

The actions of prolactin (PRL) on target cells depend on the type of prolactin receptor (PRLr) predominantly expressed, particularly whether the long PRLr isoform is expressed. The aims of this study were to determine the cellular localization and the changes in expression of long and short PRLr isoforms in sheep ovary throughout the estrous cycle. Long and short PRLrs were localized mostly in the same ovarian cells. Maximum signal intensity, particularly for long PRLrs, was found in stromal cells surrounding primordial and primary follicles, and, for both PRLrs, in granulosa cells of preantral follicles and in luteal cells. Moderate signal intensity for PRLrs was found in theca cells of preantral to ovulatory follicles, and in granulosa cells of antral follicles up to the gonadotropin-dependent stage. Decreasing immunoreactivity to PRLrs was found in granulosa cells of gonadotropin-dependent to ovulatory follicles. For long PRLrs in particular, no signal was found in mural granulosa cells of gonadotropin-dependent follicles; for both isoforms, no signal was found in most granulosa cells of ovulatory follicles. In primordial to gonadotropin-dependent follicles, cellular localization of PRLr was similar on days 0, 10 and 15 of the cycle. Oocytes consistently showed positive immunostaining for PRLrs. Comparative RT-PCR analysis of long and short PRLr expression showed that the short isoform is evenly expressed throughout the estrous cycle, whereas the expression of the long form increases at the time of estrus and decreases at mid-luteal phase and at the onset of the follicular phase. Expression of long PRLrs was greater than that of short PRLrs on day 0 of cycle; expression of both isoforms was similar on day 10 and on day 15, long PRLrs expression was lower than that of short PRLrs. Our results indicate that in sheep ovary, the maximum responsiveness to PRL might occur during the preovulatory phase of the estrous cycle.     

Morphological evaluation of bioartificial hydrogels as potential tissue engineering scaffolds

Poly(vinyl alcohol) hydrogels prepared by freeze-thawing procedure represent synthetic systems widely investigated as non-biodegradable scaffolds for tissue regeneration. In order to improve the biocompatibility properties of pure poly(vinyl alcohol) (PVA) hydrogels, blends of PVA with different biological macromolecules, such hyaluronic acid, dextran, and gelatin were prepared and used to produce bioartificial hydrogels. The porosity characteristics of these hydrogels were investigated by scanning electron microscopy and mercury intrusion porosimetry. The morphology of bioartificial hydrogels was evaluated and compared with that of pure PVA hydrogels. In particular the effect exerted by each biological component on pore size and distribution was investigated. The obtained results indicate that when a natural macromolecule is added to PVA the internal structure of the material changes. A small amount of biopolymer induces the structural elements of PVA matrix to take on a well evident lamellar appearance and an apparent preferential orientation. Comparing the results of SEM and mercury intrusion porosimetry it was concluded that hydrogels containing 20% of biological component have the most regular structure and at the same time the lowest total porosity. On the contrary samples with the highest content of natural polymer (40%) show the less regular structure and the highest total porosity.     

The raison d'être of constitutively active protein kinases: the lesson of CK2

Protein kinases generally are tightly controlled signaling molecules that are switched on only in response to specific stimuli. Exceptionally few protein kinases are constitutively active, the most striking example being provided by CK2 (formerly "casein kinase 2"). Owing to unique structural features, the catalytic activity of CK2 is constantly on, although its targeting can be deeply influenced by the association of its two catalytic subunits (alpha and/or alpha') with a dimeric non catalytic beta subunit. Constitutive activity of CK2 reflects its extraordinary pleiotropy documented by its growing list of >300 protein substrates and is consistent with emerging evidence that CK2 plays an essential role in the cell by counteracting premature and/or unscheduled apoptosis, thus ensuring cell survival under stress conditions.     

An Integrated CMOS Front-End for Optical Absolute Rotary Encoders

This paper presents a front-end circuit for optical rotary encoders. The light pulses modulated by the encoder disc are transduced into current signals, which are pre-processed and converted into digital waveforms related to the disc angular position information. The proposed front-end circuit is compensated against temperature drifts. Digitally programmable calibration is provided to account for spreads in impinging light pulse power. Measurement results on integrated prototypes are shown, demonstrating correct operation of the front-end with an optical input power from 0.5 W to 3 W up to a signal frequency of 500 kHz in a temperature range from 0 °C to 80 °C.     

Optimisation of the throttling system in a CO2 refrigerating machine

This paper is an answer to the need of finding the optimal solution for the throttling system in refrigerating machines using CO₂ as working fluid; such a solution must combine reliability, low installation cost and high energy efficiency. To this purpose, different expansion systems are compared by means of a simulation programme, including a new one, operating with a differential valve, a liquid receiver and a thermostatic valve. The typical compression refrigerating cycle performed by CO₂ involves transcritical operations and therefore the upper pressure needs to be adjusted to the optimal value, that, unlike the traditional cycle, is not determined by heat transfer. The innovative system here proposed shows an intrinsic self-adjusting capability that leads to COP values quite close to the maximum ones when a fixed suitable value of the differential pressure is chosen, even if the temperature of the secondary fluid varies to a large extent.     

Squaring down LTI systems: A geometric approach

In this paper, the problem of reducing a given LTI system into a left or right invertible one is addressed and solved with the standard tools of the geometric control theory. First, it will be shown how an LTI system can be turned into a left invertible system, thus preserving key system properties like stabilizability, phase minimality, right invertibility, relative degree and infinite zero structure. Moreover, the additional invariant zeros introduced in the left invertible system thus obtained can be arbitrarily assigned in the complex plane. By duality, the scheme of a right inverter will be derived straightforwardly. Moreover, the squaring down problem will be addressed. In fact, when the left and right reduction procedures are applied together, a system with an unequal number of inputs and outputs is turned into a square and invertible system. Furthermore, as an example it will be shown how these techniques may be employed to weaken the standard assumption of left invertibility of the plant in many optimization problems.     

Simulation of the St. Francis Dam-Break Flood

A two-dimensional (2D) simulation of flooding from the 1928 failure of St. Francis Dam in southern California is presented. The simulation algorithm solves shallow-water equations using a robust unstructured grid Godunov-type scheme designed for wetting and drying and achieves good results. Flood extent and flood travel time are predicted within 4 and 10% of observations, respectively. Representation of terrain by the mesh is identified as the dominant factor affecting accuracy, and an iterative process of mesh refinement and convergence checks is implemented to minimize errors. The most accurate predictions are achieved with a uniformly distributed Manning n = 0.02. A 50% increase in n increases travel time errors to 25% but has little effect on flood extent predictions. This highlights the challenge of a priori travel time prediction but robustness in flood extent prediction when topography is well resolved. Predictions show a combination of subcritical and supercritical flow regimes. The leading edge of the flood was supercritical in San Francisquito Canyon, but due to channel tortuosity, the wetting front reflected off canyon walls causing a transition to subcritical flow, considerably larger depths, and a standing wave in one particular reach that accounts for a 30% fluctuation in discharge. Elsewhere, oblique shocks locally increased flood depths. The 2D dam-break model is validated by its stability and accuracy, conservation properties, ability to calibrate with a physically realistic and simple resistance parametrization, and modest computational cost. Further, this study highlights the importance of a dynamic momentum balance for dam-break flood simulation.