Investigation of the effects of extracellular osmotic pressure on morphology and mechanical 1 properties of individual chondrocyte

It has been demonstrated that most cells of the body respond to osmotic pressure in a systematic manner. The disruption of the collagen network in the early stages of osteoarthritis causes an increase in water content of cartilage which leads to a reduction of pericellular osmolality in chondrocytes distributed within the extracellular environment. It is therefore arguable that an insight into the mechanical properties of chondrocytes under varying osmotic pressure would provide a better understanding of chondrocyte mechanotransduction and potentially contribute to knowledge on cartilage degeneration. In this present study, the chondrocyte cells were exposed to solutions with different osmolality. Changes in their dimensions and mechanical properties were measured over time. Atomic Force Microscopy (AFM) was used to apply load at various strain-rates and the force-time curves were logged. The thin-layer elastic model was used to extract the elastic stiffness of chondrocytes at different strain-rates and at different solution osmolality. In addition, the porohyperelastic (PHE) model was used to investigate the strain-rate dependent responses under the loading and osmotic pressure conditions. The results revealed that the hypo-osmotic external environment increased chondrocyte dimensions and reduced Young’s modulus of the cells at all strain-rates tested. In contrast, the hyper-osmotic external environment reduced dimensions and increased Young’s modulus. Moreover, by using the PHE model coupled with inverse FEA simulation, we established that the hydraulic permeability of chondrocytes increased with decreasing extracellular osmolality which is consistent with previous work in the literature. This could be due to a higher intracellular fluid volume fraction with lower osmolality.

Studies on Plant Aspartate Transcarbamylase Purification and Properties of the Enzyme from Mung-Bean (Phaseolus aureus) Seedlings

Aspartate transcarbamylase is purified from mung bean seedlings by a series of steps involving manganous sulphate treatment, ammonium sulphate fractionation, DEAE-cellulose chromatography, followed by a second ammonium sulphate fractionation and finally gel filtration on Sephadex-G 100. The enzyme is homogeneous on ultracentrifugation and on polyacrylamide gel electrophoresis. It functions optimally at 55°C. It has two pH optima, one at 8.0 and the other at 10.2. The enzyme follows Michaelis-Menten kinetics with l-aspartate as the variable substrate. However, it exhibits sigmoid saturation curves at both the pH optima when the concentration of carbamyl phosphate is varied. The enzyme is allosterically inhibited by UMP at both the pH optima. Increasing phosphorylation of the uridine nucleotide decreases the inhibitory effect. The enzyme is desensitized to inhibition by UMP on treatment with p-hydroxymercuribenzoate, gel electrophoresis indicating that the enzyme is dissociated by this treatment; the dissociated enzyme can be reassociated by treatment with 2-mercaptoethanol. The properties of the mung bean enzyme are compared with the enzyme from other sources.

Efficient simulation of unsaturated flow in heterogeneous soils using an exponential integrator

- Provided a practical variable-stepsize implementation of the exponential Euler method (EEM). - Introduced a new second-order variant of the scheme that enables the local error to be estimated at the cost of a single additional function evaluation. - New EEM implementation outperformed sophisticated implementations of the backward differentiation formulae (BDF) of order 2 and was competitive with BDF of order 5 for moderate to high tolerances.

Oral medication delivery in impaired swallowing: Thickening liquid medications for safe swallowing alters dissolution characteristics

Acetaminophen (paracetamol) is available in a wide range of oral formulations designed to meet the needs of the population across the age-spectrum, but for people with impaired swallowing, i.e. dysphagia, both solid and liquid medications can be difficult to swallow without modification. The effect of a commercial polysaccharide thickener, designed to be added to fluids to promote safe swallowing by dysphagic patients, on rheology and acetaminophen dissolution was tested using crushed immediate-release tablets in water, effervescent tablets in water, elixir and suspension. The inclusion of the thickener, comprised of xanthan gum and maltodextrin, had a considerable impact on dissolution; acetaminophen release from modified medications reached 12-50% in 30 minutes, which did not reflect the pharmacopeia specification for immediate release preparations. Flow curves reflect the high zero-shear viscosity and the apparent yield stress of the thickened products. The weak gel nature, in combination with high G’ values compared to G” (viscoelasticity) and high apparent yield stress, impact drug release. The restriction on drug release from these formulations is not influenced by the theoretical state of the drug (dissolved or dispersed), and the approach typically used in clinical practice (mixing crushed tablets into pre-prepared thickened fluid) cannot be improved by altering the order of incorporation or mixing method.

A Framework for Detecting Glaucomatous Progression in the Optic Nerve Head of an Eye Using Proper Orthogonal Decomposition

Glaucoma is the second leading cause of blindness worldwide. Often, the optic nerve head (ONH) glaucomatous damage and ONH changes occur prior to visual field loss and are observable in vivo. Thus, digital image analysis is a promising choice for detecting the onset and/or progression of glaucoma. In this paper, we present a new framework for detecting glaucomatous changes in the ONH of an eye using the method of proper orthogonal decomposition (POD). A baseline topograph subspace was constructed for each eye to describe the structure of the ONH of the eye at a reference/baseline condition using POD. Any glaucomatous changes in the ONH of the eye present during a follow-up exam were estimated by comparing the follow-up ONH topography with its baseline topograph subspace representation. Image correspondence measures of L-1-norm and L-2-norm, correlation, and image Euclidean distance (IMED) were used to quantify the ONH changes. An ONH topographic library built from the Louisiana State University Experimental Glaucoma study was used to evaluate the performance of the proposed method. The area under the receiver operating characteristic curves (AUCs) was used to compare the diagnostic performance of the POD-induced parameters with the parameters of the topographic change analysis (TCA) method. The IMED and L-2-norm parameters in the POD framework provided the highest AUC of 0.94 at 10 degrees. field of imaging and 0.91 at 15 degrees. field of imaging compared to the TCA parameters with an AUC of 0.86 and 0.88, respectively. The proposed POD framework captures the instrument measurement variability and inherent structure variability and shows promise for improving our ability to detect glaucomatous change over time in glaucoma management.

Drying characteristics and quality of grape under physical pretreatment

Grape drying is a slow and energy intensive process because the waxy peel has low permeability to moisture. Therefore, peel chemical and physical pretreatments are considered before drying in order to facilitate water diffusion. However, they cause heterogeneity in the waxes removal and problems during shelf-life. In this paper an alternative abrasive pretreatment of grape peel, for enhancing the drying rate and preserving the samples, was applied to Red Globe grapes. Convective drying experiments were carried out at 40-70 Centigrade and at 2.3 m/s air velocity. The effect of wax abrasive pretreatment on the drying kinetics and quality parameters of raisins was investigated. The results were compared with those of samples pretreated by dipping in alkaline ethyl oleate solution and untreated grapes. All the dried samples are darker than fresh one and shrunked. The samples pretreated by peel abrasion and dried at 50 centigrade showed the lowest color changes, less shrinkage and the best rehydration capacity. The drying kinetics and shrinkage curves were also analyzed using some commonly available empirical models.

Conformal Field Theory Methods for Variants of Schramm-Loewner Evolutions

This thesis consists of an introduction, four research articles and an appendix. The thesis studies relations between two different approaches to continuum limit of models of two dimensional statistical mechanics at criticality. The approach of conformal field theory (CFT) could be thought of as the algebraic classification of some basic objects in these models. It has been succesfully used by physicists since 1980's. The other approach, Schramm-Loewner evolutions (SLEs), is a recently introduced set of mathematical methods to study random curves or interfaces occurring in the continuum limit of the models. The first and second included articles argue on basis of statistical mechanics what would be a plausible relation between SLEs and conformal field theory. The first article studies multiple SLEs, several random curves simultaneously in a domain. The proposed definition is compatible with a natural commutation requirement suggested by Dubédat. The curves of multiple SLE may form different topological configurations, ``pure geometries''. We conjecture a relation between the topological configurations and CFT concepts of conformal blocks and operator product expansions. Example applications of multiple SLEs include crossing probabilities for percolation and Ising model. The second article studies SLE variants that represent models with boundary conditions implemented by primary fields. The most well known of these, SLE(kappa, rho), is shown to be simple in terms of the Coulomb gas formalism of CFT. In the third article the space of local martingales for variants of SLE is shown to carry a representation of Virasoro algebra. Finding this structure is guided by the relation of SLEs and CFTs in general, but the result is established in a straightforward fashion. This article, too, emphasizes multiple SLEs and proposes a possible way of treating pure geometries in terms of Coulomb gas. The fourth article states results of applications of the Virasoro structure to the open questions of SLE reversibility and duality. Proofs of the stated results are provided in the appendix. The objective is an indirect computation of certain polynomial expected values. Provided that these expected values exist, in generic cases they are shown to possess the desired properties, thus giving support for both reversibility and duality.

Identification of the sex pheromone of the tree infesting Cossid Moth Coryphodema tristis (Lepidoptera: Cossidae)

The cossid moth (Coryphodema tristis) has a broad range of native tree hosts in South Africa. The moth recently moved into non-native Eucalyptus plantations in South Africa, on which it now causes significant damage. Here we investigate the chemicals involved in pheromone communication between the sexes of this moth in order to better understand its ecology, and with a view to potentially develop management tools for it. In particular, we characterize female gland extracts and headspace samples through coupled gas chromatography electro-antennographic detection (GC-EAD) and two dimensional gas chromatography mass spectrometry (GCxGC-MS). Tentative identities of the potential pheromone compounds were confirmed by comparing both retention time and mass spectra with authentic standards. Two electrophysiologically active pheromone compounds, tetradecyl acetate (14:OAc) and Z9-tetradecenyl acetate (Z9-14:OAc) were identified from pheromone gland extracts, and an additional compound (Z9-14:OH) from headspace samples. We further determined dose response curves for the identified compounds and six other structurally similar compounds that are common to the order Cossidae. Male antennae showed superior sensitivity toward Z9-14:OAc, Z7-tetradecenyl acetate (Z7-14:OAc), E9-tetradecenyl acetate (E9-14:OAc), Z9-tetradecenol (Z9-14:OH) and Z9-tetradecenal (Z9-14:Ald) when compared to female antennae. While we could show electrophysiological responses to single pheromone compounds, behavioral attraction of males was dependent on the synergistic effect of at least two of these compounds. Signal specificity is shown to be gained through pheromone blends. A field trial showed that a significant number of males were caught only in traps baited with a combination of Z9-14:OAc (circa 95 of the ratio) and Z9-14:OH. Addition of 14:OAc to this mixture also improved the number of males caught, although not significantly. This study represents a major step towards developing a useful attractant to be used in management tools for C. tristis and contributes to the understanding of chemical communication and biology of this group of insects.

Estimating daily methane production in individual cattle with irregular feed intake patterns from short-term methane emission measurements

Spot measurements of methane emission rate (n = 18 700) by 24 Angus steers fed mixed rations from GrowSafe feeders were made over 3- to 6-min periods by a GreenFeed emission monitoring (GEM) unit. The data were analysed to estimate daily methane production (DMP; g/day) and derived methane yield (MY; g/kg dry matter intake (DMI)). A one-compartment dose model of spot emission rate v. time since the preceding meal was compared with the models of Wood (1967) and Dijkstra et al. (1997) and the average of spot measures. Fitted values for DMP were calculated from the area under the curves. Two methods of relating methane and feed intakes were then studied: the classical calculation of MY as DMP/DMI (kg/day); and a novel method of estimating DMP from time and size of preceding meals using either the data for only the two meals preceding a spot measurement, or all meals for 3 days prior. Two approaches were also used to estimate DMP from spot measurements: fitting of splines on a 'per-animal per-day' basis and an alternate approach of modelling DMP after each feed event by least squares (using Solver), summing (for each animal) the contributions from each feed event by best-fitting a one-compartment model. Time since the preceding meal was of limited value in estimating DMP. Even when the meal sizes and time intervals between a spot measurement and all feeding events in the previous 72 h were assessed, only 16.9% of the variance in spot emission rate measured by GEM was explained by this feeding information. While using the preceding meal alone gave a biased (underestimate) of DMP, allowing for a longer feed history removed this bias. A power analysis taking into account the sources of variation in DMP indicated that to obtain an estimate of DMP with a 95% confidence interval within 5% of the observed 64 days mean of spot measures would require 40 animals measured over 45 days (two spot measurements per day) or 30 animals measured over 55 days. These numbers suggest that spot measurements could be made in association with feed efficiency tests made over 70 days. Spot measurements of enteric emissions can be used to define DMP but the number of animals and samples are larger than are needed when day-long measures are made.

Open-circuit potential-time transient of the magnesium anode

On interrupting polarisation, the magnesium anode exhibits a negative overshoot in potential followed by a slow recovery to a steady state value. A model has been proposed to explain the opencircuit potential-time transient in terms of a spontaneous passivation of the metal and the consequent changes in the corrosion potential. Theoretical expressions have been derived for the timedependence of the open-circuit electrode potential. Calculated, potential-time curves thus obtained are in qualitative agreement with experimental data. A possible application of this phenomenon to develop non-destructive quality control tests of Mg, Li and Al-based dry cells has been pointed out.

Studies On Hydrogen Bonds: Part IV. Proposed Working Criteria for Assessing Qualitative Strength of Hydrogen Bonds

The data obtained in the earlier parts of this series for the donor and acceptor end parameters of N-H. O and O-H. O hydrogen bonds have been utilised to obtain a qualitative working criterion to classify the hydrogen bonds into three categories: "very good" (VG), "moderately good" (MG) and weak (W). The general distribution curves for all the four parameters are found to be nearly of the Gaussian type. Assuming that the VG hydrogen bonds lie between 0 and ± la, MG hydrogen bonds between ± 1 and ± 2, W hydrogen bonds beyond ± 2 (where is the standard deviation), suitable cut-off limits for classifying the hydrogen bonds in the three categories have been derived. These limits are used to get VG and MG ranges for the four parameters 1 and θ (at the donor end) and ± and ± (at the acceptor end). The qualitative strength of a hydrogen bond is decided by the cumulative application of the criteria to all the four parameters. The criterion has been further applied to some practical examples in conformational studies such as α-helix and can be used for obtaining suitable location of hydrogen atoms to form good hydrogen bonds. An empirical approach to the energy of hydrogen bonds in the three categories has also been presented.

Turbulence measurements in an equilibrium axisymmetric wall jet

This paper reports measurements of turbulent quantities in an axisymmetric wall jet subjected to an adverse pressure gradient in a conical diffuser, in such a way that a suitably defined pressure-gradient parameter is everywhere small. Self-similarity is observed in the mean velocity profile, as well as the profiles of many turbulent quantities at sufficiently large distances from the injection slot. Autocorrelation measurements indicate that, in the region of turbulent production, the time scale of ν fluctuations is very much smaller than the time scale of u fluctuations. Based on the data on these time scales, a possible model is proposed for the Reynolds stress. One-dimensional energy spectra are obtained for the u, v and w components at several points in the wall jet. It is found that self-similarity is exhibited by the one-dimensional wavenumber spectrum of $\overline{q^2}(=\overline{u^2}+\overline{v^2}+\overline{w^2})$, if the half-width of the wall jet and the local mean velocity are used for forming the non-dimensional wavenumber. Both the autocorrelation curves and the spectra indicate the existence of periodicity in the flow. The rate of dissipation of turbulent energy is estimated from the $\overline{q^2}$ spectra, using a slightly modified version of a previously suggested method.

Studies on Rat Ovarian Receptors for Lutropin (Luteinizing Hormone): Applicability of radioimmunoassay to measure lutropin bound to receptors.

Measurement of receptor-bound unlabelled physiologically active lutropin (luteinizing hormone, LH) was possible by a modified radioimmunoassay. The conventional radioimmunoassayconducted at 4°C was inadequate, whereas the modified assay performed at 37'C could measure receptor-bound lutropin. The radioimmunoassay at 37'C takes only 36h for completion compared with 5-7 days at 4°C. The sensitivity and range of dose-response curves are, however, unaltered. The validity of the technique was established by a number of criteria.

Vibration of generally orthotropic skew plates

Vibration problem of generally orthotropic plates with particular attention to plates of skew geometry is studied. The formulation is based on orthotropic plate theory with arbitrary orientation of the principal axes of orthotropy. The boundary conditions considered are combinations of simply supported, clamped, and free-edge conditions. Approximate solution for frequencies and modes is obtained by the Ritz method using products of appropriate beam characteristic functions as admissible functions. The variation of frequencies and modes with orientation of the axes of orthotropy is examined for different skew angles and boundary conditions. Features such as "crossings" and "quasi-degeneracies" of the frequency curves are found to occur with variation of the orientation of the axes of orthotropy for a given geometry of the skew plate. It is also found that for each combination of skew angle and side ratio, a particular orientation of the axes gives the highest value for the fundamental frequency of the plate.

Vibration of skew plates

The vibration problems of skew plates with different edge conditions involving simple support and clamping have been considered by using the variational method of Ritz, a double series of beam characteristic functions being employed appropriate to the combination of the edge conditions. Natural frequencies and modes of vibration have been obtained for different combinations of side ratio and skew angle. These detailed studies reveal several interesting features concerning the frequency curves and nodal patterns. The results presented should, in addition, be of considerable value and practical significance in design applications.