Experimental methods for studying drug uptake in the head and brain

A number of techniques have been developed to study the disposition of drugs in the head and, in particular, the role of the blood-brain barrier (BBB) in drug uptake. The techniques can be divided into three groups: in-vitro, in-vivo and in-situ. The most suitable method depends on the purpose(s) and requirements of the particular study being conducted. In-vitro techniques involve the isolation of cerebral endothelial cells so that direct investigations of these cells can be carried out. The most recent preparations are able to maintain structural and functional characteristics of the BBB by simultaneously culturing endothelial cells with astrocytic cells,The main advantages of the in-vitro methods are the elimination of anaesthetics and surgery. In-vivo methods consist of a diverse range of techniques and include the traditional Brain Uptake Index and indicator diffusion methods, as well as microdialysis and positron emission tomography. In-vivo methods maintain the cells and vasculature of an organ in their normal physiological states and anatomical position within the animal. However, the shortcomings include renal acid hepatic elimination of solutes as well as the inability to control blood flow. In-situ techniques, including the perfused head, are more technically demanding. However, these models have the ability to vary the composition and flow rate of the artificial perfusate. This review is intended as a guide for selecting the most appropriate method for studying drug uptake in the brain.

Experimental tests of quantum nonlinear dynamics in atom optics

Cold atoms in optical potentials provide an ideal test bed to explore quantum nonlinear dynamics. Atoms are prepared in a magneto-optic trap or as a dilute Bose-Einstein condensate and subjected to a far detuned optical standing wave that is modulated. They exhibit a wide range of dynamics, some of which can be explained by classical theory while other aspects show the underlying quantum nature of the system. The atoms have a mixed phase space containing regions of regular motion which appear as distinct peaks in the atomic momentum distribution embedded in a sea of chaos. The action of the atoms is of the order of Planck's constant, making quantum effects significant. This tutorial presents a detailed description of experiments measuring the evolution of atoms in time-dependent optical potentials. Experimental methods are developed providing means for the observation and selective loading of regions of regular motion. The dependence of the atomic dynamics on the system parameters is explored and distinct changes in the atomic momentum distribution are observed which are explained by the applicable quantum and classical theory. The observation of a bifurcation sequence is reported and explained using classical perturbation theory. Experimental methods for the accurate control of the momentum of an ensemble of atoms are developed. They use phase space resonances and chaotic transients providing novel ensemble atomic beamsplitters. The divergence between quantum and classical nonlinear dynamics is manifest in the experimental observation of dynamical tunnelling. It involves no potential barrier. However a constant of motion other than energy still forbids classically this quantum allowed motion. Atoms coherently tunnel back and forth between their initial state of oscillatory motion and the state 180 out of phase with the initial state.

User dyads in software testing: bypassing the need for expert observers

This paper arises out of a research study into the online help facilities provided in popular software applications such as word processors. Its particular focus is on experimental methods of evaluating the effectiveness and usability of those facilities. Focus groups, questionnaires, and online surveys had already been used in other phases of the study, but it was judged that these approaches would be unsuitable for measuring effectiveness and usability because they are susceptible to respondents' subjectivity. Direct observation of people working on set word-processing tasks was ruled out initially because of a lack of trained observers; it would have taken too long for the investigator to observe a large enough sample by himself. Automatic recording of users' actions was also rejected, as it would have demanded equipment and/or software that was not available and seemed too expensive to acquire. The approach and techniques described here were an attempt to overcome these difficulties by using observers drawn from the same population of students that provided the test subjects; as a by-product, this may also have enhanced the acceptability (and hence possibly the validity) of the experiments by reducing the exam pressure perceived by participants.

Modelling cell lifespan and proliferation: is likelihood to die or to divide independent of age?

In cell lifespan studies the exponential nature of cell survival curves is often interpreted as showing the rate of death is independent of the age of the cells within the population. Here we present an alternative model where cells that die are replaced and the age and lifespan of the population pool is monitored until a, steady state is reached. In our model newly generated individual cells are given a determined lifespan drawn from a number of known distributions including the lognormal, which is frequently found in nature. For lognormal lifespans the analytic steady-state survival curve obtained can be well-fit by a single or double exponential, depending on the mean and standard deviation. Thus, experimental evidence for exponential lifespans of one and/or two populations cannot be taken as definitive evidence for time and age independence of cell survival. A related model for a dividing population in steady state is also developed. We propose that the common adoption of age-independent, constant rates of change in biological modelling may be responsible for significant errors, both of interpretation and of mathematical deduction. We suggest that additional mathematical and experimental methods must be used to resolve the relationship between time and behavioural changes by cells that are predominantly unsynchronized.

Evaluation of parameters effective in FRP shear strengthening of RC beams using FE method

Shear strengthening is required when an RC beam is found deficient in shear, or when its shear capacity falls below its flexural capacity after flexural strengthening. A recent technique for the shear strengthening of RC beams is to provide additional FRP web reinforcement, commonly in the form of bonded external FRP strips/sheets. Over the last few years, several experimental studies have been conducted on this new strengthening technique, which has established its effectiveness. While experimental methods of investigation are extremely useful in obtaining information about the composite behaviour of FRP and reinforced concrete, the use of numerical models such as the one presented in this paper helps in developing a good understanding of the behaviour at lower costs. In the study presented in this paper, ANSYS finite element program is used to examine the response of beams strengthened in shear by FRPs. The FE model is calibrated against test results performed at the University of Kentucky. Once validated, the model is used to examine the influence of fibre orientation, compressive strength of concrete, area of tensile and compressive reinforcements, and amount and distance between stirrups on the strength and ductility of FRP strengthened beam.

Experimental study of the quantum driven pendulum and its classical analog in atom optics

We present experimental results for the dynamics of cold atoms in a far detuned amplitude-modulated optical standing wave. Phase-space resonances constitute distinct peaks in the atomic momentum distribution containing up to 65% of all atoms resulting from a mixed quantum chaotic phase space. We characterize the atomic behavior in classical and quantum regimes and we present the applicable quantum and classical theory, which we have developed and refined. We show experimental proof that the size and the position of the resonances in phase space can be controlled by varying several parameters, such as the modulation frequency, the scaled well depth, the modulation amplitude, and the scaled Planck’s constant of the system. We have found a surprising stability against amplitude noise. We present methods to accurately control the momentum of an ensemble of atoms using these phase-space resonances which could be used for efficient phase-space state preparation.

Diseño hidráulico de vertedores escalonados con pendientes moderadas: Metodología basada en un estudio experimental

Durante las últimas tres décadas el interés y diversidad en el uso de canales escalonados han aumentado debido al desarrollo de nuevas técnicas y materiales que permiten su construcción de manera rápida y económica (Concreto compactado con rodillo CCR, Gaviones, etc.). Actualmente, los canales escalonados se usan como vertedores y/o canales para peces en presas y diques, como disipadores de energía en canales y ríos, o como aireadores en plantas de tratamiento y torrentes contaminados. Diversos investigadores han estudiado el flujo en vertedores escalonados, enfocándose en estructuras de gran pendiente (  45o) por lo que a la fecha, el comportamiento del flujo sobre vertedores con pendientes moderadas (  15 a 30o) no ha sido totalmente comprendido. El presente artículo comprende un estudio experimental de las propiedades físicas del flujo aire-agua sobre canales escalonados con pendientes moderadas, típicas en presas de materiales sueltos. Un extenso rango de gastos en condiciones de flujo rasante se investigó en dos modelos experimentales a gran escala (Le = 3 a 6): Un canal con pendiente 3.5H:1V (  16o) y dos alturas de escalón distintas (h = 0.1 y 0.05 m) y un canal con pendiente 2.5H:1V (  22o) y una altura de escalón de h = 0.1 m. Los resultados incluyen un análisis detallado de las propiedades del flujo en vertedores escalonados con pendientes moderadas y un nuevo criterio de diseño hidráulico, el cual está basado en los resultados experimentales obtenidos. English abstract: Stepped chutes have been used as hydraulic structures since antiquity, they can be found acting as spillways and fish ladders in dams and weirs, as energy dissipators in artificial channels, gutters and rivers, and as aeration enhancers in water treatment plants and polluted streams. In recent years, new construction techniques and materials (Roller Compacted Concrete RCC, rip-rap gabions, etc.) together with the development of the abovementioned new applications have allowed cheaper construction methods, increasing the interest in stepped chute design. During the last three decades, research in stepped spillways has been very active. However, studies prior to 1993 neglected the effect of free-surface aeration. A number of studies have focused since on steep stepped chutes (  45o) but the hydraulic performance of moderate-slope stepped channels is not yet totally understood. This study details an experimental investigation of physical air-water flow properties down moderate slope stepped spillways conducted in two laboratory models: the first model was a 3.15 m long stepped chute with a 15.9o slope comprising two interchangeable step heights (h = 0.1 m and h = 0.05 m); the second model was a 3.3 m long, stepped channel with a 21.8o slope (h = 0.1 m). A broad range of discharges within transition and skimming flow regimes was investigated. Measurements were conducted using a double tip conductivity probe. The study provides new, original insights into air-water stepped chute flows not foreseen in prior studies and presents a new design criterion for chutes with moderate slopes based on the experimental results.

An initial assessment of policies for saving a rare Australian glider: Experimental results, economics and ecology

Reviews the ecological status of the mahogany glider and describes its distribution, habitat and abundance, life history and threats to it. Three serial surveys of Brisbane residents provide data on the knowledge of respondents about the mahogany glider. The results provide information about the attitudes of respondents to the mahogany glider, to its conservation and relevant public policies and about variations in these factors as the knowledge of participants of the mahogany glider alters. Similarly data is provided and analysed about the willingness to pay of respondents to conserve the mahogany glider. Population viability analysis is applied to estimate the required habitat area for a minimum viable population of the mahogany glider to ensure at least a 95% probability of its survival for 100 years. Places are identified in Queensland where the requisite minimum area of critical habitat can be conserved. Using the survey results as a basis, the likely willingness of groups of Australians to pay for the conservation of the mahogany glider is estimated and consequently their willingness to pay for the minimum required area of its habitat. Methods for estimating the cost of protecting this habitat are outlined. Australia-wide benefits seem to exceed the costs. Establishing a national park containing the minimum viable population of the mahogany glider is an appealing management option. This would also be beneficial in conserving other endangered wildlife species. Therefore, additional economic benefits to those estimated on account of the mahogany glider itself can be obtained.

Improved methods for predicting individual leaf area and leaf senescence in maize (Zea mays)

The ability to predict leaf area and leaf area index is crucial in crop simulation models that predict crop growth and yield. Previous studies have shown existing methods of predicting leaf area to be inadequate when applied to a broad range of cultivars with different numbers of leaves. The objectives of the study were to (i) develop generalised methods of modelling individual and total plant leaf area, and leaf senescence, that do not require constants that are specific to environments and/or genotypes, (ii) re-examine the base, optimum, and maximum temperatures for calculation of thermal time for leaf senescence, and (iii) assess the method of calculation of individual leaf area from leaf length and leaf width in experimental work. Five cultivars of maize differing widely in maturity and adaptation were planted in October 1994 in south-eastern Queensland, and grown under non-limiting conditions of water and plant nutrient supplies. Additional data for maize plants with low total leaf number (12-17) grown at Katumani Research Centre, Kenya, were included to extend the range in the total leaf number per plant. The equation for the modified (slightly skewed) bell curve could be generalised for modelling individual leaf area, as all coefficients in it were related to total leaf number. Use of coefficients for individual genotypes can be avoided, and individual and total plant leaf area can be calculated from total leaf number. A single, logistic equation, relying on maximum plant leaf area and thermal time from emergence, was developed to predict leaf senescence. The base, optimum, and maximum temperatures for calculation of thermal time for leaf senescence were 8, 34, and 40 degrees C, and apply for the whole crop-cycle when used in modelling of leaf senescence. Thus, the modelling of leaf production and senescence is simplified, improved, and generalised. Consequently, the modelling of leaf area index (LAI) and variables that rely on LAI will be improved. For experimental purposes, we found that the calculation of leaf area from leaf length and leaf width remains appropriate, though the relationship differed slightly from previously published equations.

Methods for prediction of peptide binding to MHC molecules: A comparative study

Background: A variety of methods for prediction of peptide binding to major histocompatibility complex (MHC) have been proposed. These methods are based on binding motifs, binding matrices, hidden Markov models (HMM), or artificial neural networks (ANN). There has been little prior work on the comparative analysis of these methods. Materials and Methods: We performed a comparison of the performance of six methods applied to the prediction of two human MHC class I molecules, including binding matrices and motifs, ANNs, and HMMs. Results: The selection of the optimal prediction method depends on the amount of available data (the number of peptides of known binding affinity to the MHC molecule of interest), the biases in the data set and the intended purpose of the prediction (screening of a single protein versus mass screening). When little or no peptide data are available, binding motifs are the most useful alternative to random guessing or use of a complete overlapping set of peptides for selection of candidate binders. As the number of known peptide binders increases, binding matrices and HMM become more useful predictors. ANN and HMM are the predictive methods of choice for MHC alleles with more than 100 known binding peptides. Conclusion: The ability of bioinformatic methods to reliably predict MHC binding peptides, and thereby potential T-cell epitopes, has major implications for clinical immunology, particularly in the area of vaccine design.

Special issue on methods and learning in functional MRI: Introductory remarks

This special issue represents a further exploration of some issues raised at a symposium entitled “Functional magnetic resonance imaging: From methods to madness” presented during the 15th annual Theoretical and Experimental Neuropsychology (TENNET XV) meeting in Montreal, Canada in June, 2004. The special issue’s theme is methods and learning in functional magnetic resonance imaging (fMRI), and it comprises 6 articles (3 reviews and 3 empirical studies). The first (Amaro and Barker) provides a beginners guide to fMRI and the BOLD effect (perhaps an alternative title might have been “fMRI for dummies”). While fMRI is now commonplace, there are still researchers who have yet to employ it as an experimental method and need some basic questions answered before they venture into new territory. This article should serve them well. A key issue of interest at the symposium was how fMRI could be used to elucidate cerebral mechanisms responsible for new learning. The next 4 articles address this directly, with the first (Little and Thulborn) an overview of data from fMRI studies of category-learning, and the second from the same laboratory (Little, Shin, Siscol, and Thulborn) an empirical investigation of changes in brain activity occurring across different stages of learning. While a role for medial temporal lobe (MTL) structures in episodic memory encoding has been acknowledged for some time, the different experimental tasks and stimuli employed across neuroimaging studies have not surprisingly produced conflicting data in terms of the precise subregion(s) involved. The next paper (Parsons, Haut, Lemieux, Moran, and Leach) addresses this by examining effects of stimulus modality during verbal memory encoding. Typically, BOLD fMRI studies of learning are conducted over short time scales, however, the fourth paper in this series (Olson, Rao, Moore, Wang, Detre, and Aguirre) describes an empirical investigation of learning occurring over a longer than usual period, achieving this by employing a relatively novel technique called perfusion fMRI. This technique shows considerable promise for future studies. The final article in this special issue (de Zubicaray) represents a departure from the more familiar cognitive neuroscience applications of fMRI, instead describing how neuroimaging studies might be conducted to both inform and constrain information processing models of cognition.

Selective down-regulation of the astrocyte glutamate transporters GLT-1 and GLAST within the medial thalamus in experimental wernicke's encephalopathy

Although earlier studies on thiamine deficiency have reported increases in extracellular glutamate concentration in the thalamus, a vulnerable region of the brain in this disorder, the mechanism by which this occurs has remained unresolved. Treatment with pyrithiamine, a central thiamine antagonist, resulted in a 71 and 55% decrease in protein levels of the astrocyte glutamate transporters GLT-1 and GLAST, respectively, by immunoblotting in the medial thalamus of day 14 symptomatic rats at loss of righting reflexes. These changes occurred prior to the onset of convulsions and pannecrosis. Loss of both GLT-1 and GLAST transporter sites was also confirmed in this region of the thalamus at the symptomatic stage using immunohistochemical methods. In contrast, no change in either transporter protein was detected in the non-vulnerable frontal parietal cortex. These effects are selective; protein levels of the astrocyte GABA transporter GAT-3 were unaffected in the medial thalamus. In addition, astrocyte-specific glial fibrillary acidic protein (GFAP) content was unchanged in this brain region, suggesting that astrocytes are spared in this disorder. Loss of GLT-1 or GLAST protein was not observed on day 12 of treatment, indicating that down-regulation of these transporters occurs within 48 h prior to loss of righting reflexes. Finally, GLT-1 content was positively correlated with levels of the neurofilament protein alpha -internexin, suggesting that early neuronal drop-out may contribute to the down-regulation of this glutamate transporter and subsequent pannecrosis. A selective, focal loss of GLT-1 and GLAST transporter proteins provides a rational explanation for the increase in interstitial glutamate levels, and may play a major role in the selective vulnerability of thalamic structures to thiamine deficiency-induced cell death.

A potential role for the analytical ultracentrifuge in the experimental measurement of protein valence

A method is described whereby sedimentation velocity is combined with equilibrium dialysis to determine the net charge (valence) of a protein by using chromate as an indicator ion for assessing the extent of the Donnan redistribution of small ions. The procedure has been used in experiments on bovine serum albumin under slightly alkaline conditions (pH 8.0, I 0.05) to illustrate its application to a system in which the indicator ion and protein both bear net negative charge and on lysozyme under slightly acidic conditions (pH 5.0, I 0.10) to illustrate the situation where chromate is a counterion. (C) 2001 Elsevier Science.

Vibration instability in rolling mills: Modeling and experimental results

Models for the occurrence of the vibrational instability during rolling known as third octave chatter are presented and discussed. An analysis of rolling mill chatter was performed for the purpose of identifying characteristics of the vibrations and to determine any dependency on the rolling schedule. In particular, a stability criterion for the critical rolling speed is used to predict the maximum rolling speed without chatter instability on schedules from a 5 stand tandem mill rolling thin steel product. The results correlate well with measurements of critical speed occurring on the mill using a vibration monitor: This research provides significant insights into the chatter phenomena and has been used to investigate control methods for suppression of the instability.