Gene/Protein Disease Symptom Drug Enzyme Compound
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Poly(N-isopropylacrylamide) (PNIPAM) microgel particles dispersed in water have been used as a matrix for the polymerization of a hydrophobic monomer, N-methylpyrrole (MPy). The presence of poly(MPy) (PMPy) within the dried composite particles has been confirmed using electron paramagnetic resonance (EPR) measurements which show a characteristic free-radical signal at g=2.007. Electron microscopy data (TEM) show that the composite PNIPAM-xPMPy particles have a "raspberry-like" morphology. (The value for x represents the volume percent of MPy added during synthesis with respect to the total microgel volume.) PCS data indicate that PMPy occupies the majority of the collapsed composite particle volume. The maximum value of x consistent with colloid stability for PNIPAM-xPMPy dispersions is 4.5%. Higher values of x result in coagulation due to interparticle bridging by PMPy. Variable temperature PCS measurements of the PNIPAM-xPMPy dispersions have been used to study the thermally induced collapse of the composite particles. The extent of collapse becomes less with increasing values for x. The embedded PMPy particles restrict the extent of PNIPAM network contraction. The stability of the PNIPAM-4.5PMPy dispersions was investigated by means of turbidity measurements using aqueous 0.1 M NaCl solution. The upper critical flocculation temperatures (UCFT) for PNIPAM and PNIPAM-4.5PMPy dispersions were identical (38-39 degrees C). The flocculation observed was found to be fully reversible. The composite dispersion stability in the absence of salt was attributed to electrosteric stabilization afforded by the PNIPAM matrix. These results indicate that PNIPAM microgel particles may have application as a matrix for the polymerization of hydrophobic monomers in water. Copyright 2000 Academic Press.
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PMID:Microgel Particles as a Matrix for Polymerization: A Study of Poly(N-isopropylacrylamide)-Poly(N-methylpyrrole) Dispersions. 1065 27

A one-pot, soft-chemistry, surfactant-assisted co-assembly approach to prepare La(1-x)Sr(x)MnO(3) (LSM)/Y(2)O(3)-stabilized ZrO(2) (YSZ) nanocomposites for use as solid oxide fuel cell (SOFC) cathodes has been investigated. This material with sub-hundred nanometer grain sizes for each phase is the first such nanocomposite where aqueous-based precursors of each component are incorporated in a single synthetic step. This approach utilizes the co-assembly of an anionic yttrium/zirconium acetatoglycolate gel, cetyltrimethylammonium bromide as the cationic surfactant template, and inorganic La, Mn, and Sr salts under alkaline aqueous conditions. The resulting as-synthesized product is an amorphous mesostructured organic/inorganic composite, which is transformed to a mesoporous inorganic oxide with nanocrystalline YSZ walls upon calcination. Calcination to temperatures above 600 degrees C lead to collapse of the mesopores followed by further crystallization of the nanocrystalline YSZ phase and a final crystallization of the LSM perovskite phase above 1000 degrees C. Both the fully crystalline LSM/YSZ and the mesoporous intermediate phase have been investigated for phase homogeneity by TEM energy-dispersive X-ray spectroscopy (EDX) mapping and spot analysis which confirm the dispersion of LSM within a YSZ matrix at the nanometer scale. Impedance spectroscopy analysis of LSM/YSZ nanocomposite electrodes demonstrate a low polarization resistance of around 0.2 omega cm(2) with an activation energy (E(a)) as low as 1.42 eV. Cathodic polarization studies show stable current densities over a 40 h test demonstration.
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PMID:Lanthanum strontium manganite/yttria-stabilized zirconia nanocomposites derived from a surfactant assisted, co-assembled mesoporous phase. 1270 68

Different sizes of nanotubes of poly(2-methoxy-5-(n-hexadecyloxy)-p-phenylene vinylene)(MH-PPV) have been fabricated at the air/water interface by compressing a monolayer of MH-PPV beyond its collapse point, and their structural characteristics were studied by means of TEM, AFM, SAXRD, IRRAS.
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PMID:Nanotubes of poly(phenylene vinylene) derivative at the air/water interface. 1526 69

We investigate the conformation of long polyelectrolyte chains attached to colloidal latex particles by cryogenic transmission electron microscopy (cryo-TEM). The dense grafting of the polyelectrolyte chains ("polyelectrolyte brush") leads to a confinement of the counterions and a concomitantly high osmotic pressure within the polyelectrolyte layer attached to the core particles. Cryo-TEM has provided first model-independent direct proof for the strong stretching of the polyelectrolyte chains by direct visualization. If salt is added, cryo-TEM clearly shows how chains collapse because of the strong screening of the electrostatic interaction. Moreover, the analysis of interacting particles by cryo-TEM shows that the polyelectrolyte chains retract at close contact. Hence, we demonstrate how cryo-TEM can be used to analyze directly the spatial structure of polyelectrolyte brushes in situ.
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PMID:High elongation of polyelectrolyte chains in the osmotic limit of spherical polyelectrolyte brushes: a study by cryogenic transmission electron microscopy. 1599 64

Beta-lactamase acquisition is the most prevalent basis for Gram-negative bacteria resistance to the beta-lactam antibiotics. The mechanism used by the most common class A Gram-negative beta-lactamases is serine acylation followed by hydrolytic deacylation, destroying the beta-lactam. The ab initio quantum mechanical/molecular mechanical (QM/MM) calculations, augmented by extensive molecular dynamics simulations reported herein, describe the serine acylation mechanism for the class A TEM-1 beta-lactamase with penicillanic acid as substrate. Potential energy surfaces (based on approximately 350 MP2/6-31+G calculations) reveal the proton movements that govern Ser70 tetrahedral formation and then collapse to the acyl-enzyme. A remarkable duality of mechanism for tetrahedral formation is implicated. Following substrate binding, the pathway initiates by a low energy barrier (5 kcal mol(-1)) and an energetically favorable transfer of a proton from Lys73 to Glu166, through the catalytic water molecule and Ser70. This gives unprotonated Lys73 and protonated Glu166. Tetrahedral formation ensues in a concerted general base process, with Lys73 promoting Ser70 addition to the beta-lactam carbonyl. Moreover, the three-dimensional potential energy surface also shows that the previously proposed pathway, involving Glu166 as the general base promoting Ser70 through a conserved water molecule, exists in competition with the Lys73 process. The existence of two routes to the tetrahedral species is fully consistent with experimental data for mutant variants of the TEM beta-lactamase.
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PMID:Ab initio QM/MM study of class A beta-lactamase acylation: dual participation of Glu166 and Lys73 in a concerted base promotion of Ser70. 1626 3

Dynamic light scattering (DLS) measurement provides an effective way to investigate the formation of nanotube of gamma-cyclodextrin (gamma-CD) induced by N,N'-diphenylbenzidine (DPB) in water. With the combination of steady-state fluorescence and fluorescence anisotropy experiments, it was found that for alpha- and beta-CD, only 1:2 (guest:host) inclusion complexes were formed and for gamma-CD, cyclodextrin nanotube was formed involving 16 gamma-CD units at maximum. The pH effect studies with both DLS and fluorescence anisotropy measurements indicated that the hydrogen bonding between neighboring CDs was necessary to the formation of cyclodextrin nanotube. In the temperature experiment, we found that the nanotube of DPB-gamma-CD could exist stably at relatively high temperatures and the transition point for structural collapse was estimated to be around 54 degrees C. The aggregation states of both gamma-CD itself and DPB-gamma-CD nanotube were observed with TEM.
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PMID:Investigation on gamma-cyclodextrin nanotube induced by N,N'-diphenylbenzidine molecule. 1635 9

The serous (poison) cutaneous glands of the Pacific tree-frog Hyla regilla were induced to release their product by 10(-3)M nor-epinephrine stimulation. After discharge structural and ultrastructural features of the cutaneous glands involved in release were observed. Furthermore, the discharged product, consisting of discrete, secretory granules, was collected and processed for transmission electron microscope analysis. As indicated by patterns found in the myoepithelium encircling the syncytial secretory unit, gland discharge is caused by contraction of the peripheral myocytes. Muscle cell compression dramatically affects the syncytium and results in degenerative changes, including expulsion of the secretory unit nuclei. Therefore, the structural collapse in depleted glands has been ascribed to the mechanical activity performed by the myoepithelium during discharge, rather than cytoplasm involution described in conventional, holocrine glands. TEM investigation revealed that the secretory granules collected after discharge maintain their peculiar traits: they consist of recurrent patterns of thin subunits, acquired during serous maturation and provided with remarkable structural stability.
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PMID:Serous cutaneous glands of the Pacific tree-frog Hyla regilla (Anura, Hylidae): patterns of secretory release induced by nor-epinephrine. 1642 75

The formation of a complex between an anionic spherical polyelectrolyte brush (SPB) and the cationic surfactant cetyltrimethylammonium bromide (CTAB) is investigated. The SPB consists of long chains of the strong polyelectrolyte poly(styrene sulfonate) (PSS), which are bound chemically to a solid poly(styrene) core of 56 nm in radius. The SPB are dispersed in water, and the ionic strength is adjusted by addition of NaBr. The resulting complexes are investigated in dilute solution by dynamic light scattering, by electrophoretic light scattering, and by cryogenic transmission electron microscopy (cryo-TEM). The formation of the complex between the SPB and the surfactant can be monitored by a strong shrinking of the surface layer when adding CTAB to dilute suspensions (0.01 wt %) and by a decrease of the effective charge of the complexes. Complex formation starts at CTAB concentrations lower than the critical micelle concentration of this surfactant. If the ratio r of the charges on the SPB to the charge of the added surfactant is exceeding unity, the particles start to flocculate. Cryo-TEM images of the complexes at r = 0.6 measured in salt-free solution show that the surface layer composed of the PSS chains and the adsorbed CTAB molecules is partially collapsed: A part of the chains form a dense surface layer while another part of the chains or aggregates thereof are still sticking out. This can be deduced from the cryo-TEM micrographs as well as from the hydrodynamic radius, which is still of appreciable magnitude. The 1:1 complex (r = 1.0) exhibits a fully collapsed layer formed by the PSS chains and CTAB. If the complex is formed in the presence of 0.05 M NaBr, r = 0.6 leads to globular structures directly attached to the surface of the core particles. All structures seen in the cryo-TEM images can be explained by a collapse transition of the surface layer brought about by the hydrophobic attraction between the polyelectrolyte chains that became partially hydrophobic through adsorption of CTAB.
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PMID:Binding of oppositely charged surfactants to spherical polyelectrolyte brushes: a study by cryogenic transmission electron microscopy. 1731 35

The conformational stability and kinetics of refolding and unfolding of the W290F mutant of TEM-1 beta-lactamase have been determined as a function of guanidinium chloride concentration. The activity and spectroscopic properties of the mutant enzyme did not differ significantly from those of the wild type, indicating that the mutation has only a very limited effect on the structure of the protein. The stability of the folded protein is reduced, however, by 5-10 kJ mol-1 relative to that of the molten globule intermediate (H), but the values of the folding rate constants are unchanged, suggesting that Trp-290 becomes organized in its nativelike environment only after the rate-limiting step; i.e., the C-terminal region of the enzyme folds very late. In contrast to the significant increase in fluorescence intensity seen in the dead time (3-4 ms) of refolding of the wild-type protein, no corresponding burst phase was observed with the mutant enzyme, enabling the burst phase to be attributed specifically to the C-terminal Trp-290. This residue is suggested to be buried in a nonpolar environment from which it has to escape during subsequent folding steps. With both proteins, fast early collapse leads to a folding intermediate in which the C-terminal region of the polypeptide chain is trapped in a non-native structure, consistent with a nonhierarchical folding process.
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PMID:TEM-1 beta-lactamase folds in a nonhierarchical manner with transient non-native interactions involving the C-terminal region. 1817 Oct 85

Vesicles composed of sodium oleate (NaO) and monoolein (MO) are adequate candidates for drug nanoencapsulation and controlled release due to their stability and perceived biocompatibility. The object of the present study is to design hydrogels based on those anionic vesicles and polymers of both non-associative and associative type. The selected macromolecules were k-carrageenan (KC), carboxymethyl cellulose (CMC) and hydrophobically modified carboxymethyl cellulose (HMCMC). While the polymer-vesicle association was probed by rheology, the influence of the polymer on the vesicle stability was monitored by cryo-TEM and calorimetric measurements. The effects of the polymer on the rheological properties of surfactant aggregate solutions clearly depend on the polymer type: the storage moduli of the polymer-vesicle mixtures, compared to the vesicles alone, increases around 2 orders of magnitude if the polymer is non-associative and 4 orders of magnitude if the macromolecule is of associative type. As the vesicles are added, the non-associative polymer networks tend to be disrupted, while the networks formed by associative polymer get more robust. These observations can be explained by fundamental changes in electrostatic/hydrophobic interactions: vesicles entrapped in KC networks convert the polysaccharide in a highly charged entity and favor high electrostatic repulsions between the chains; this encourages network collapse. The opposite picture is experienced in HMCMC systems, i.e., such network is stabilized by the presence of vesicles. This is ascribed to the enhanced hydrophobic association, compensating the electrostatic repulsions between vesicles and polymer chains.
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PMID:Gelation of charged bio-nanocompartments induced by associative and non-associative polysaccharides. 1862 14


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