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Biomedical subjects

A Faraone

Publications and source records attributed to A Faraone.

At least 19 recordsLinked to original sources

Experimental evidence of fragile-to-strong dynamic crossover in DNA hydration water.

We used high-resolution quasielastic neutron scattering spectroscopy to study the single-particle dynamics of water molecules on the surface of hydrated DNA samples. Both H(2)O and D(2)O hydrated samples were measured. The contribution of scattering from DNA is subtracted out by taking the difference of the signals between the two samples. The measurement was made at a series of temperatures from 270 down to 185 K. The relaxing-cage model was used to analyze the quasielastic spectra. This allowed us to extract a Q-independent average translational relaxation time of water molecules as a function of temperature. We observe clear evidence of a fragile-to-strong dynamic crossover (FSC) at T(L)=222+/-2 K by plotting log versus T. The coincidence of the dynamic transition temperature T(c) of DNA, signaling the onset of anharmonic molecular motion, and the FSC temperature T(L) of the hydration water suggests that the change of mobility of the hydration water molecules across T(L) drives the dynamic transition in DNA.

DNA↗

Observation of fragile-to-strong dynamic crossover in protein hydration water.

At low temperatures, proteins exist in a glassy state, a state that has no conformational flexibility and shows no biological functions. In a hydrated protein, at temperatures greater-- similar 220 K, this flexibility is restored, and the protein is able to sample more conformational substates, thus becoming biologically functional. This "dynamical" transition of protein is believed to be triggered by its strong coupling with the hydration water, which also shows a similar dynamic transition. Here we demonstrate experimentally that this sudden switch in dynamic behavior of the hydration water on lysozyme occurs precisely at 220 K and can be described as a fragile-to-strong dynamic crossover. At the fragile-to-strong dynamic crossover, the structure of hydration water makes a transition from predominantly high-density (more fluid state) to low-density (less fluid state) forms derived from the existence of the second critical point at an elevated pressure.

Animals↗

The fragile-to-strong dynamic crossover transition in confined water: nuclear magnetic resonance results.

By means of a nuclear magnetic resonance experiment, we give evidence of the existence of a fragile-to-strong dynamic crossover transition (FST) in confined water at a temperature T(L)=223+/-2 K. We have studied the dynamics of water contained in 1D cylindrical nanoporous matrices (MCM-41-S) in the temperature range 190-280 K, where experiments on bulk water were so far hampered by crystallization. The FST is clearly inferred from the T dependence of the inverse of the self-diffusion coefficient of water (1D) as a crossover point from a non-Arrhenius to an Arrhenius behavior. The combination of the measured self-diffusion coefficient D and the average translational relaxation time tau(T), as measured by neutron scattering, shows the predicted breakdown of Stokes-Einstein relation in deeply supercooled water.

Journal Article↗

Simulation of exposure and SAR estimation for adult and child heads exposed to radiofrequency energy from portable communication devices.

The level and distribution of radiofrequency energy absorbed in a child's head during the use of a mobile phone compared to those in an adult head has been a controversial issue in recent years. It has been suggested that existing methods that are used to determine specific absorption rate (SAR) and assess compliance with exposure standards using an adult head model may not adequately account for potentially higher levels of exposure in children due to their smaller head size. The present study incorporates FDTD computations of locally averaged SAR in two different anatomically correct adult and child head models using the IEEE standard (Std. C95.3-2002) SAR averaging algorithm. The child head models were obtained by linear scaling of the adult head model to replicate the conditions of previous studies reported in the literature and also by transforming the different adult head models based on data on the external shapes of children's heads. The tissue properties of the adult and corresponding child head models were kept the same. In addition, modeling and experimental measurements were made using three spheres filled with a tissue-equivalent mixture to approximate heads of increasing size. Results show that the peak local average SAR over 1 g and 10 g of tissue and the electromagnetic energy penetration depths are about the same in all of the head models under the same exposure conditions. When making interlaboratory comparisons, the model and the SAR averaging algorithm used must be standardized to minimize controversy.

Adult↗

Fragile-to-strong liquid transition in deeply supercooled confined water.

Confining water in lab synthesized nanoporous silica matrices MCM-41-S with pore diameters of 18 and 14 A, we have been able to study the molecular dynamics of water in deeply supercooled states, down to 200 K. Using quasielastic neutron scattering and analyzing the data with the relaxing cage model, we determined the temperature variation of the average translational relaxation time and its Q-dependence. We find a clear evidence of an abrupt change of the relaxation time behavior at T approximately equal to 225 K, which we interpreted as the predicted fragile-to-strong liquid-liquid transition.

Journal Article↗

Dynamics of supercooled water in mesoporous silica matrix MCM-48-S.

Using three different quasielastic neutron spectrometers with widely different resolutions, we have been able to study the microscopic translational and rotational dynamics of water, in a mesoporous silica matrix MCM-48-S, from T=300 K to 220 K, with a single consistent model. We formulated our fitting routine using the relaxing cage model. Thus, from the fit of the experimental data, we extracted the fraction of water bound to the surface of the pore, the characteristic relaxation times of the long-time translational and rotational decays, the stretch exponent describing the shape of the relaxation processes, and the power exponent determining the Q-dependence of the translational relaxation time. A tremendous slowing down of the rotational relaxation time, as compared to the translational one, has been observed.

Journal Article↗

Estimation of the SAR in the human head and body due to radiofrequency radiation exposure from handheld mobile phones with hands-free accessories.

It was reported by others that hands-free accessories increase the absorption of RF energy in a human head compared to a handset alone. The results of this study show that the opposite is observed when proper dosimetric methods are employed. It is pointed out that for correct estimation of the exposure level it is necessary to use appropriate physical and experimental models and measurement instrumentation, following internationally recommended standards. The human phantoms used for measurements involving the hands-free accessories should include the torso; i.e., measurements should not be performed on the head phantom alone. This has a significant impact on the results because the RF energy coupled into the leads of hands-free accessories is strongly attenuated by the body. Numerical simulations using the Finite-Difference Time-Domain (FDTD) method and experimental measurements with a miniature electric-field probe are in good agreement and show a decrease, not an increase, in RF energy exposure in the human head from hands-free accessories.

Cell Phone↗

Rotational dynamics of hydration water in dicalcium silicate by quasielastic neutron scattering.

Quasielastic neutron scattering (QENS) has been used to investigate the single-particle dynamics of interfacial water in dicalcium silicate (C2S)/water paste. Our previous neutron-scattering studies on interfacial water have focused attention on the translational dynamics of the center of mass of water molecules. In this paper, we have collected QENS data on a wider range of wave-vector transfer so that both translational and rotational motions of water molecules are detected. The data have been analyzed by models for translation and rotation we recently proposed for supercooled water. The evolution of the parameters describing the relaxational dynamics of water embedded in the C2S matrix is given at temperature T=303 K as a function of the curing time.

Journal Article↗

Observation of a re-entrant kinetic glass transition in a micellar system with temperature-dependent attractive interaction.

We detect in a tri-block co-polymer micellar system an ergodic-to-nonergodic-to-ergodic transition, as a function of temperature, in a range of concentrations, by photon correlation measurements. The shear viscosity is also shown to jump two order of magnitude at these transition temperatures. Surprisingly, the structure factor as measured by small angle neutron scattering shows a marked narrowing at the structural arrest state. Rationalization of these results with the existence of an attractive branch in the phase diagram of an attractive colloid system predicted by mode coupling theory is made.

Journal Article↗

Morphological and biochemical variations of Haemophilus influenzae type b induced by pH and temperature changes.

Haemophilus influenzae type b ATCC 10211 was cultured at different temperatures (25 degrees C-49 degrees C) and pH values (5.7-8.7) either in liquid or semisolid medium. Morphological variations of individual cells were noted by optical microscopy depending upon the conditions of growth. At higher temperatures filaments were produced whereby the length of individual cells increased compared to cultures grown at 37 degrees C. Filaments were also observed at lower pH values. Culture conditions also affected colonial morphology. At low pH values colonies had an enhanced lobulated contour and were more wrinkly and rougher than at higher pH. The changes in cellular and colonial morphology were correlated with distinct outer membrane protein profiles. The changes in temperature and pH did not affect identification of the microorganism by the API system.

Bacterial Outer Membrane Proteins↗

Characterization of glutathione transferase from Xanthomonas campestris.

A single form of glutathione transferase (Xc-GST-4.5) having an isoelectric point at pH 4.5 was resolved from Xanthomonas campestris cytosol by affinity chromatography and isoelectric focusing. HPLC,N-terminal amino acid sequence, and SDS-PAGE analyses indicate that Xc-GST-4.5 is composed of two identical subunits, each with a molecular mass of 22 kDa. As indicated by its substrate specificity, immunological reactivity, and CD spectra, as well as by its N-terminal amino acid sequence, Xc-GST-4.5 appears to be distinct from the other bacterial glutathione transferases, Pm-GST-6.0 and Sm-GST-7.3, previously purified from the cytosolic fraction of Proteus mirabilis and Serratia marcescens. Xc-GST-4.5 also appears to be distinct from the GST so far purified from other sources.

Amino Acid Sequence↗

Purification of a GSH-affinity binding protein from Bacteroides fragilis devoid of glutathione transferase activity.

An affinity binding protein from the cytosolic fraction of Bacteroides fragilis was purified by using epoxy activated-Sepharose 6B resin immobilized with GSH or with hexyl-GSH. This protein showed a subunit molecular mass (22 kDa) similar to that of glutathione transferase purified from Proteus mirabilis (22.5 kDa). However, the affinity binding protein of Bacteroides fragilis, unlike the GSH-affinity binding protein of Proteus mirabilis, was devoid of the capacity to conjugate GSH to the most commonly used glutathione transferase substrates. The GSH-affinity binding protein of Bacteroides fragilis was also antigenically different from the GSH-affinity bound protein of Proteus mirabilis. It was concluded that the anaerobic microorganism is not able to express glutathione transferase even though it contains a GSH-affinity binding protein with a structural characteristic reminiscent of aerobic glutathione transferase.

Anaerobiosis↗

Purification and characterization of a novel glutathione transferase from Serratia marcescens.

Four forms of glutathione transferase were resolved from the cytosol of Serratia marcescens CIP 6755 by GSH-affinity chromatography followed by isoelectric focusing. The major isoenzyme, named Sm-GST-7.3, is composed of two subunits each with a molecular mass of 22 kDa and has an isoelectric point at pH 7.3. Sm-GST-7.3, appears to be distinct from Pm-GST-6.0, previously characterized from Proteus mirabilis AF 2924 as indicated by its substrate specificity, immunological reactivity, subunit molecular mass as well as by its N-terminal amino acid sequence. None of the antisera raised against a number of human, rat and mouse GSTs cross-reacted with Sm-GST-7.3 indicating major structural differences between them and bacterial GST. This is further supported by the fact that the N-terminal sequence of Sm-GST-7.3 also differs significantly from the known sequences of mammalian GSTs of alpha, mu and pi classes. In addition, comparison with the known N-terminal amino acid sequences of helminth, plant and insect GSTs demonstrate that the latter enzymes are distantly related (less than 25% identity) to the Sm-GST-7.3. Immunoblotting experiments performed with antisera raised against Sm-GST-7.3 indicate that a GST immunologically identical to Sm-GST-7.3 is present in a number of other bacterial strains. All together the results obtained suggest that Sm-GST-7.3 is distinct from any known GST, including microbial and mammalian GSTs.

Amino Acid Sequence↗

Glutathione transferase in bacteria: subunit composition and antigenic characterization.

The presence of glutathione transferase (GST; EC 2.5.1.18) in Escherichia coli ATCC 25922, E. coli ATCC 25422, Proteus vulgaris ATCC 8427, Pseudomonas aeruginosa ATCC 27853, Klebsiella oxytoca CIP 666, K. oxytoca AF 101, Enterobacter cloacae CIP 6085, Serratia marcescens CIP 6755, and Proteus mirabilis AF 2924 was investigated. Using 1-chloro-2,4-dinitrobenzene as substrate, GST activity was found in the glutathione-(GSH-)affinity-purified fraction of all strains tested. SDS-PAGE analysis of GSH-affinity-purified enzyme indicated that the GSTs of all these bacteria are dimers of two identical subunits of Mr about 22,500. Rabbit antiserum directed against the major isoenzyme present in Proteus mirabilis AF 2924, Pm-GST-6.0, was used to investigate the antigenic properties of bacterial GSTs. Western blot analysis indicated that a GST antigenically identical to Pm-GST-6.0 is present in Enterobacter cloacae CIP 6085, Escherichia coli ATCC 25422 and Proteus vulgaris ATCC 8427, but absent in Escherichia coli ATCC 25922, Klebsiella oxytoca CIP 666, K. oxytoca AF 101 and Serratia marcescens CIP 6755. The presence of Pm-GST-6.0, but not mammalian GST, increased the MIC values of amikacin, ampicillin, cefotaxime, cephalothin and nalidixic acid for E. coli ATCC 25922. It is suggested that bacterial GST may represent a defense against the effects of antibiotics.

Anti-Bacterial Agents↗

Purification and characterization of three forms of glutathione transferase from Proteus mirabilis.

Three forms of glutathione transferase (GST) with pI values of 6.0, 6.4 and 7.3 were isolated from Proteus mirabilis AF 2924 by glutathione-affinity chromatography followed by isoelectric focusing, and their structural, kinetic and immunological properties were investigated. Upon SDS/polyacrylamide-slab-gel electrophoresis, all forms proved to be composed of two subunits of identical (22,500) Mr. GST-6.0 and GST-6.4 together account for about 95% of the total activity, whereas GST-7.3 is present only in trace amounts. Extensive similarities have been found between GST-6.0 and GST-6.4. These include subunit molecular mass, amino acid composition, substrate specificities and immunological characteristics. GST-7.3 also cross-reacted (non-identity) with antisera raised against bacterial GST-6.0. None of the antisera raised against a number of human, rat and mouse GSTs cross-reacted with the bacterial enzymes, indicating major structural differences between them and the mammalian GSTs. This conclusion is further supported by c.d. spectra.

Amino Acids↗

Preliminary studies on the effect of glutathione S-transferase from Providencia stuartii on the antimicrobial activity of different antibiotics.

Glutathione S-transferase has been isolated and purified from a Providencia stuartii CH 114 strain. The effect of the enzyme on the antimicrobial activity of amikacin, cefotaxime, cephalexin, ampicillin, nalidixic acid, and ofloxacin was tested. The efficiency of all antibiotics tested, except nalidixic acid and ofloxacin, is relevantly decreased by the presence of glutathione S-transferase in the medium culture, as proved by the increased value of MIC. The effect of glutathione S-transferase on the drugs is not significantly affected by the addition of exogenous glutathione. The possible mechanism of action of the enzyme is also discussed.

Anti-Bacterial Agents↗

beta 1-selective adrenoceptor antagonists. 1. Synthesis and beta-adrenergic blocking activity of a series of binary (aryloxy)propanolamines.

A series of binary (aryloxy)propanolamines has been prepared and examined in vitro and in vivo for beta-adrenoreceptor blocking activity. These symmetrical compounds consist of two (S)-(phenyloxy)propanolamine pharmacophores coupled through alkylenedioxy or poly(oxyethylenedioxy) linking units of varying lengths. Examples of such binary compounds linked through the 2,2', 3,3', and 4,4' positions in the aromatic rings of the pharmacophores have been prepared. In vitro and in vivo test data indicate that the 2,2' compounds tend to be selective beta 2-adrenergic blocking agents, the 4,4' binaries tend to be selective beta 1-blocking agents, and those compounds with 3,3' linkages exhibit intermediate selectivities. One of the 4,4'-linked binary compounds, 4s, exhibited potent, cardioselective beta-blockade in vivo, which was of short duration and was accompanied by a prolonged tachycardia.

Adrenergic beta-Antagonists↗