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Biomedical subjects

M P Patel

Publications and source records attributed to M P Patel.

At least 19 recordsLinked to original sources

Dimensional changes of alginate dental impression materials.

The weight loss and corresponding dimensional changes of two dental alginate impression materials have been studied. The weight loss kinetics indicate this to be a diffusion controlled process, but with a boundary condition at the surface of the concentration decreasing exponentially with time. This is in marked contrast to most desorption processes, where the surface concentration becomes instantaneously zero. The appropriate theory has been developed for an exponential boundary condition, and its predictions compared with experimental data; the agreement was satisfactory. The diffusion coefficients for two thicknesses of the same material were not identical as predicted by theory; the possible reasons for this are discussed.

Alginates↗

Mycobacterium tuberculosis mycothione reductase: pH dependence of the kinetic parameters and kinetic isotope effects.

The recent identification of the enzyme in Mycobacterium tuberculosis that catalyzes the NADPH-dependent reduction of the unique low molecular weight disulfide mycothione, mycothione reductase, has led us to examine the mechanism of catalysis in greater detail. The pH dependence of the kinetic parameters V and V/K for NADPH, NADH, and an active analogue of mycothione disulfide, des-myo-inositol mycothione disulfide, has been determined. An analysis of the pH profiles has allowed the tentative assignment of catalytically significant residues crucial to the mechanism of disulfide reduction, namely, the His444-Glu449 ion pair and Cys39. Solvent kinetic isotope effects were observed on V and V/K(DIMSSM), yielding values of 1.7 +/- 0.2 and 1.4 +/- 0.2, respectively, but not on V/K(NADPH). Proton inventory studies (V versus mole fraction of D(2)O) were linear, indicative of a single proton transfer in a solvent isotopically sensitive step. Steady-state primary deuterium kinetic isotope effects on V have been determined using NADPH and NADH, yielding values of 1.27 +/- 0.03 and 1.66 +/- 0.14, respectively. The pre-steady-state primary deuterium kinetic isotope effect on enzyme reduction has values of 1.82 +/- 0.04 and 1.59 +/- 0.06 for NADPH and NADH, respectively. The steady-state primary deuterium kinetic isotope effect using NADH coincide with that obtained under single turnover conditions, suggesting the complete expression of the intrinsic primary kinetic isotope effect. Rapid reaction studies on the reductive half-reaction using NADPH and NADH yielded maximal rates of 129 +/- 2 and 20 +/- 1 s(-1), respectively, while similar studies of the oxidation of the two-electron reduced enzyme by mycothiol disulfide yielded a maximum rate of 190 +/- 10 s(-1). These data suggest a unique flavoprotein disulfide mechanism in which the rate of the oxidative half-reaction is slightly faster than the rate of the reductive half-reaction.

Deuterium↗

Nuclear magnetic imaging of an osmotic water uptake and delivery process.

The water absorption of NH4F-doped silicone cylinders was studied by spin-echo Fourier nuclear magnetic resonance imaging, in conjunction with gravimetric (mass uptake) and ionic concentration (release of NH4F) measurements. The addition of NH4F greatly increases the water uptake as the inclusions are surrounded by droplets of water when wetted by water diffusing in the polymer. These droplets then expand due to the osmotic pressure difference between the internal droplet and external solution; this expansion is opposed by an elastic restraining force. The rate of ingress into the polymer will depend on the relative chemical potential of water associated with the polymer and the droplet, as well as the diffusion coefficient. Later stages of the absorption process are dominated by the droplets. The dilution of the droplets (from water diffusing into the droplets) slowly reduces the chemical potential driving the process. Hence, the later stages occur without any notable concentration gradient but a slight chemical potential gradient. If the droplet expansion exceeds that which can be restrained by the material, failure around the droplet occurs resulting in the release of the salt and the solution within the droplet.

Ammonium Compounds↗

The effect of two hydrophilic monomers on the water uptake of a heterocyclic methacrylate system.

The room temperature polymerising system poly(ethyl methacrylate) (PEM)/tetrahydrofurfuryl methcrylate (THFM) has been modified by replacing some of the THFM by hydroxyethyl methacrylate (HEMA) and hydroxypropyl methacrylate (HPM), respectively. In both cases, the equilibrium uptake of the parent system is reduced substantially, in spite of the hydrophilic nature of these monomers. The effect is less with HPM. Corresponding to these decreases in uptake are substantial increases in the diffusion coefficients. This points to changes from a cluster-dominated process, to a more continuum-based process in the dual monomer systems. Addition of chicken serum albumin to these systems increases water uptake. At higher levels of HEMA addition, there is a substantial increase in polymerisation exotherm.

Absorption↗

A polymeric system for the intra-oral delivery of an anti-fungal agent.

Oral candidal infections are often persistent and intractable and thus the aim of this study was to develop a polymeric sustained release device to improve the topical treatment of these infections. A self curing system based on poly(ethyl methacrylate) and tetrahydrofurfuryl methacrylate (PEM/THFM) was used with chlorhexidine diacetate (CX) added at levels between 0 and 12% w/w. Water uptake by the device was assessed gravimetrically and CX release measured by UV spectrometry. Anti candidal activity was established by culturing azole sensitive and resistant strains of Candida albicans in the presence of the polymeric delivery device with and without CX. Candidal growth was measured by turbidimetry or surviving colony-forming unit (CFU) formation. There was an initial high release of CX over 24 h followed by a slow diffusion up to 7 days. CX inhibited candidal growth and survival markedly in vitro, with the test samples showing less than 0.5 x 10(-7) CFU/ml compared to controls (3-4 x 10(-7) CFU/ml). These results indicate the potential of a chlorhexidine containing PEM/THFM polymeric system in the treatment of persistent candidal infections.

Administration, Oral↗

Release of recombinant human bone morphogenetic protein-2 from heterocyclic methacrylate polymer systems.

The release of recombinant human bone morphogenetic protein-2 (rhBMP-2) from three room temperature polymerising methacrylate systems has been studied. These all contained poly(ethyl methacrylate) powder, but the monomer liquids comprised, respectively, tetrahydrofurfuryl methacrylate (THFM), 90/10 THFM/hydroxyethyl methacrylate (HEMA), and 70/30 THFM/ HEMA. In all cases, rhBMP-2 was released, but the addition of 10% HEMA accelerated release (a nine-fold increase in diffusion coefficient); a further increase to 30% HEMA had no additional effect. For most of the release process, a diffusion process operated, although the early stages were not well defined. At the end of the 15 day period, the release, respectively, for the PEM/THFM, PEM:90/10 THFM/HEMA and PEM:70/30 THFM/HEMA systems was 596, 878 and 923 ng (i.e. up to 92% of the rhBMP-2 added).

Biocompatible Materials↗

Isoprene-styrene copolymer elastomer and tetrahydrofurfuryl methacrylate mixtures for soft prosthetic applications.

Novel elastomer/methacrylate systems have been developed for potential soft prosthetic applications. Mixtures of varying compositions of an isoprene-styrene copolymer elastomer and tetrahydrofurfuryl methacrylate (SIS/THFMA) formed one-gel systems and were heat cured with a peroxide initiator. The blends were characterised in terms of sorption in deionised water and simulated body fluids (SBF), tensile properties and viscoelastic parameters of storage modulus and tan delta, as well as glass transition temperatures using dynamic mechanical analysis (DMA). DMA data gave two distinct peaks in tan delta, a lower temperature transition due to the isoprene phase in SIS and one at high temperature thought to be a combination of THFMA and the styrene phase in SIS. The tensile data showed a clear phase inversion within the mid range compositions changing from plastic to elastomeric behaviour. The sorption studies in deionised water showed a two stage uptake with an initial Fickian region that was linear to t 1/2 followed by a droplet growth/clustering system. The slope of the linear region was dependent on the composition ratio. The extent of overall uptake was osmotically dependent as all materials equilibrated at a much lower uptake in SBF. The diffusion coefficients were found to be concentration dependent.

Absorption↗

Replantation of the penis: a patient report.

Replantation of the penis is an unusual case in this country and it is unlikely that most plastic and reconstructive surgeons or urologists will see one during their career. A successful repair of a self-inflicted amputation of the penis is presented. The unique anatomy of the penis pertinent to replantation is reviewed, and current concepts and recommendations in performing replantation of the penis are presented.

Adult↗

Expression, purification, and characterization of Mycobacterium tuberculosis mycothione reductase.

Mycothione reductase from the human pathogen Mycobacterium tuberculosis has been cloned, expressed in Mycobacterium smegmatis, and purified 145-fold to homogeneity in 43% yield. Amino acid sequence alignment of mycothione reductase with the functionally homologous glutathione and trypanothione reductase indicates conservation of the catalytically important redox-active disulfide, histidine-glutamate ion pair, and regions involved in binding both the FAD cofactor and the substrate NADPH. The homogeneous 50 kDa subunit enzyme exists as a homodimer and is NADPH-dependent and highly specific for the structurally unique low-molecular mass disulfide, mycothione, exhibiting Michaelis constants of 8 and 73 microM for NADPH and mycothione, respectively. HPLC analysis indicated the presence of 1 mol of bound FAD per monomer as the cofactor exhibiting an absorption spectrum with a lambda(max) at 462 nm with an extinction coefficient of 11 300 M(-)(1) cm(-)(1). The reductive titration of the enzyme with NADH indicates the presence of a charge-transfer complex of one of the presumptive catalytic thiolates and FAD absorbing at ca. 530 nm. Reaction with serially truncated mycothione and other disulfides and pyridine nucleotide analogues indicates a strict minimal disulfide substrate requirement for the glucosamine moiety of mycothione. The enzyme exhibits bi-bi ping-pong kinetics with both disulfide and quinone substrates. Transhydrogenase activity is observed using NADH and thio-NADP(+), confirming the kinetic mechanism. We suggest mycothione reductase as the newest member of the class I flavoprotein disulfide reductase family of oxidoreductases.

Alkylation↗

3He ion-beam analysis of water uptake and drug delivery.

3He ion-beam analysis utilises a micronuclear reaction analysis (of deuterium and carbon) and microparticle induced X-ray emission (in this case silicon and chlorine), to determine an elemental map of the surface of a sample. This study used D(2)O to model the behaviour of water in poly(tetrahydrofurfuryl methacrylate-polyethyl methacrylate) and chlorhexidine diacetate doped silicone elastomers. The poly(tetrahydrofurfuryl methacrylate-polyethyl methacrylate) systems demonstrated an initial Fickian absorption process (diffusion coefficient 1.1 x 10(-11) m(2)s(-1)) which indicated the 2-stage nature of the polymer's absorption kinetics. The doped silicone samples demonstrated an osmotic mechanism for the controlled release of drugs, with correlation between the D(2)O and the chlorhexidine diacetate inclusions increasing during the experiment. The technique proved valuable in the analysis of delivery polymers and will undoubtedly have further applications in the development of drug delivery systems.

Chlorhexidine↗

The water uptake of poly(tetrahydrofurfuryl methacrylate).

Poly(tetrahydrofurfuryl methacrylate) possesses some unique characteristics with respect to its biocompatibility and behaviour in water. The water uptake is high (>70%) and very slow (over 3 yr), but the material remains rigid throughout the process. The mechanism behind the uptake is in two stages; an initial Fickian stage, then as the matrix approaches saturation (about 3 wt%) a second clustering mechanism takes over. The rate of uptake of the second stage of the uptake is controlled by creep (or stress relaxation), and the chemical potential driving the uptake from clustering of the furfuryl rings of the polymer. If clustering or the creep is prevented (by appropriate co-polymerisation) the polymer behaves in an ideal, Fickian manner.

Absorption↗

The epidemiology, anatomy, physiology, and treatment of erectile dysfunction in chronic renal failure patients.

Erectile dysfunction (ED) is an associated morbidity for men with chronic renal failure. An understanding of the epidemiology, anatomy, physiology, and treatment options for ED can greatly improve the quality of life for men with chronic renal failure. There are psychological and physiological causes for erectile dysfunction. Once the key features leading to the patient's loss of potency have been identified, appropriate treatment plans can be instituted, often with successful outcomes. The purpose of this article is to assist the nephrology interdisciplinary team in the management of ED by reviewing possible causes, available studies, and treatment options for their patients.

Erectile Dysfunction↗

Water sorption behaviour of polymeric systems based on tetrahydrofurfuryl methacrylate.

Earlier research has described the water absorption behaviour, drug release and biological properties of a room temperature polymerizing system based on poly(ethyl methacrylate) (PEM) powder and tetrahydrofurfuryl methacrylate (THFM) monomer. This work has been extended, with respect to water sorption behaviour, by replacing the monomer to various extents with hydroxyethyl methacrylate (HEMA), and poly(ethyl methacrylate) by ethyl methacrylate (EM)-THFM copolymers. Replacing the THFM with HEMA, and gelling with PEM, increased the diffusion coefficient progressively. The replacement of PEM by EM-THFM copolymers, when gelled with THFM monomer, substantially reduced equilibrium water uptake, and increased diffusion coefficients. However, with HEMA monomer, equilibrium uptake was unaffected, but the diffusion coefficient decreased with increasing THFM content of the copolymer. This is due to a complex interaction of THFM cross-linking the copolymer, and the effect of EM on the water uptake. Heat polymerizing the PEM-THFM system reduced equilibrium uptake and the diffusion coefficient, compared with the room temperature polymerizing system; this could reflect molecular weight differences.

Journal Article↗

Enterococcus faecalis glutathione reductase: purification, characterization and expression under normal and hyperbaric O2 conditions.

Glutathione reductase is found ubiquitously in eukaryotes and Gram-negative bacteria, and plays a significant role in bacterial defense against oxidative stress. Glutathione reductase from the Gram-positive bacterium Enterococcus faecalis was purified to homogeneity using anion exchange, hydrophobic interaction, and affinity chromatography. The homogeneous 49-kDa enzyme contained 1 mol bound FAD per subunit. The determined N-terminal amino acid sequence of the E. faecalis enzyme displays significant identity with glutathione reductases from other Gram-negative and Gram-positive bacteria, as well as yeast and human erythrocyte reductases. The kinetic mechanism is ping-pong, and the determined kinetic parameters exhibited by the E. faecalis glutathione reductase are similar to those found for glutathione reductases from yeast, Escherichia coli, and human erythrocyte. A two-fold increased expression of glutathione reductase activity and a three-fold induction of glutathione peroxidase activity were observed under hyperbaric O2 growth conditions without a corresponding change in the total glutathione and soluble thiol content. The difference in the expression of the enzyme, and its cognate substrate's intracellular concentration, under these conditions suggest that the gene encoding glutathione reductase is responsive to oxygen concentration, but that the genes encoding the glutathione synthesizing enzymes are not linked to an oxygen-sensitive promoter.

Amino Acid Sequence↗

Fluoride ion release from two methacrylate polymer systems.

The release of fluoride ions from two room-temperature polymerising systems containing sodium and potassium fluoride, respectively, has been studied. The polymer systems comprised poly(ethyl methacrylate) powder (PEM), with tetrahydrofurfuryl methacrylate (THFM), and n-butyl methacrylate (nBM), respectively. The water uptake of these systems was drastically increased by the presence of fluorides, the increase being much higher with the PEM/THFM system. In both cases, uptake was a monotonic function of the molarity of the fluoride added. The uptake process was in general non-Fickian. However, for all systems, the fluoride release process was Fickian, and diffusion coefficients could be calculated. The amount of fluoride released, and the diffusion coefficients obtained, appeared unrelated to the extent of water uptake of the parent polymers.

Absorption↗

Heterocyclic methacrylates for clinical applications-further studies of water sorption.

The room temperature polymerizing system comprising poly(ethyl methacrylate)-tetra hydrofurfuryl methacrylate (PEM/THFMA) has potential in orthopaedic and dental applications, and earlier work has shown it to have unusual water absorption characteristics. This aspect has been studied in further detail, by studying the water absorption behaviour from some biological solutions, and the effect of the addition of an antibiotic (gentamicin). For comparison purposes, a parallel system whereby tetrahydrofuryl methacrylate was replaced by hydroxyethyl methacrylate (PEM/HEMA), was studied. In the case of PEM/THFMA, water uptake was substantially reduced when absorption was carried out from solutions (from about 30% in water to about 1.5% in solutions of higher concentrations), and the corresponding diffusion coefficient increased (by a factor of several hundred). The addition of gentamicin increased uptake, but the extent of increase also decreased in solutions. It was concluded that uptake was related to the osmolarity of the external solution, and also on the presence of osmotic sites within the polymer; hence the uptake process appears to be governed by chemical potential considerations. At the higher uptakes, there was evidence of water clusters. In marked contrast, the uptake by the PEM/HEMA system was independent of the osmolarity of the external solutions, presumably due to the hydrophilic nature of HEMA.

Journal Article↗

Evaluation of accuracy and reproducibility of E test for susceptibility testing of Streptococcus pneumoniae to penicillin, cefotaxime, and ceftriaxone.

We evaluated the reproducibility with which technologists perform and interpret the E test (AB Biodisk, North America, Inc., Piscataway, N.J.) for determining the susceptibility of Streptococcus pneumoniae to penicillin, cefotaxime, and ceftriaxone. Four technologists prepared E test assays to test 124 isolates of S. pneumoniae. Each technologist then interpreted the results of the E test blinded to the interpretation of the other technologists. In addition, E test results were compared with the reference method of broth microdilution. Intraobserver and interobserver agreement were assessed by use of the kappa statistic. Interpretation of the E test and broth microdilution results showed substantial to excellent agreement, with kappa values ranging from 0.878 to 0.987. Compared with broth microdilution, no very major errors and only four major errors were made with the E test. Most minor errors with penicillin and ceftriaxone occurred for isolates with intermediate or high-level resistance, whereas for cefotaxime the minor errors were more evenly distributed between susceptible and intermediate resistance and between intermediate and high-level resistance. These results indicate that there is good agreement between technologists for the interpretation of the E test when testing the susceptibility of S. pneumoniae to penicillin, cefotaxime, and ceftriaxone and that the results of the E test agree with those of broth microdilution.

Cefotaxime↗

Extrarenal Wilms' tumour--a case report and review of literature.

Extra renal Wilm's tumour (EWT) is an exceedingly rare entity and needs to fulfill certain criteria. Of the 48 cases reported, only about 36 have been found to be convincingly proved. We present one more well proven case of EWT and have extensively reviewed the literature. The diagnostic, staging and management guidelines are discussed. Amongst the reported cases, the behaviour of the tumour appears to parallel that of similarly staged "intra renal" Wilm's tumour. Hence the staging and the management protocols of "intra renal" Wilm's tumour can well be applied to tumour of extra renal location albeit with some modifications.

Abdominal Neoplasms↗