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At least 433 records · Page 24Linked to original sources

PubMed2004

Annexin A1 interaction with a zwitterionic phospholipid monolayer: a fluorescence microscopy study.

We present the results of a fluorescence microscopy study of the interaction of annexin A1 with dipalmitoylphosphatidylcholine (DPPC) monolayers as a function of the lipid monolayer phase and the pH of the aqueous subphase. We show that annexin A1-DPPC interaction depends strongly on the domain structure of the DPPC monolayer and only weakly on the subphase pH. Annexin A1 is found to be line active, with preferential adsorption at phase boundaries. Also, annexin A1 is found to form networks in the presence of a domain structure in the monolayer. Our results point toward an important contribution of the unique N-terminal domain to the organization of the protein at the interface.

1,2-Dipalmitoylphosphatidylcholine↗

PubMed2005

Quantum dot bioconjugates for imaging, labelling and sensing.

One of the fastest moving and most exciting interfaces of nanotechnology is the use of quantum dots (QDs) in biology. The unique optical properties of QDs make them appealing as in vivo and in vitro fluorophores in a variety of biological investigations, in which traditional fluorescent labels based on organic molecules fall short of providing long-term stability and simultaneous detection of multiple signals. The ability to make QDs water soluble and target them to specific biomolecules has led to promising applications in cellular labelling, deep-tissue imaging, assay labelling and as efficient fluorescence resonance energy transfer donors. Despite recent progress, much work still needs to be done to achieve reproducible and robust surface functionalization and develop flexible bioconjugation techniques. In this review, we look at current methods for preparing QD bioconjugates as well as presenting an overview of applications. The potential of QDs in biology has just begun to be realized and new avenues will arise as our ability to manipulate these materials improves.

Biosensing Techniques↗

PubMed2006

Crystal structure of the SENP1 mutant C603S-SUMO complex reveals the hydrolytic mechanism of SUMO-specific protease.

SUMO (small ubiquitin-related modifier)-specific proteases catalyse the maturation and de-conjugation processes of the sumoylation pathway and modulate various cellular responses including nuclear metabolism and cell cycle progression. The active-site cysteine residue is conserved among all known SUMO-specific proteases and is not substitutable by serine in the hydrolysis reactions demonstrated previously in yeast. We report here that the catalytic domain of human protease SENP1 (SUMO-specific protease 1) mutant SENP1C(C603S) carrying a mutation of cysteine to serine at the active site is inactive in maturation and de-conjugation reactions. To further understand the hydrolytic mechanism catalysed by SENP1, we have determined, at 2.8 A resolution (1 A = 0.1 nm), the X-ray structure of SENP1C(C603S)-SUMO-1 complex. A comparison of the structure of SENP2-SUMO-1 suggests strongly that SUMO-specific proteases require a self-conformational change prior to cleavage of peptide or isopeptide bond in the maturation and de-conjugation processes respectively. Moreover, analysis of the interface of SENP1 and SUMO-1 has led to the identification of four unique amino acids in SENP1 that facilitate the binding of SUMO-1. By means of an in vitro assay, we further demonstrate a novel function of SENP1 in hydrolysing the thioester linkage in E1-SUMO and E2-SUMO complexes. The results disclose a new mechanism of regulation of the sumoylation pathway by the SUMO-specific proteases.

Amino Acid Sequence↗

PubMed2003

The contribution of prothrombotic disorders to peri- and neonatal ischemic stroke.

Prothrombotic disorders are believed to be important contributors to the etiology of peri- and neonatal arterial ischemic stroke and sinovenous thrombosis, which may lead to life-long disability. This article reviews hematological issues unique to the perinatal period, including: the significance of the placenta as the interface between maternal and fetal circulations; normal changes in the coagulation system of mothers during and just after gestation; and the significance of prothrombotic disorders in the mother and/or fetus. Other possible maternal and neonatal contributors to peri- and neonatal stroke are discussed, including: infection, pre-eclampsia, diabetes, and drug use in the mother; and infection, dehydration, complex congenital heart disease, extracorporeal membrane oxygenation, and catheter placement in the neonate. Possible approaches to preventing and treating perinatal and neonatal stroke are presented.

Brain Ischemia↗

PubMed2006

Surface gap solitons.

We put forward the existence of surface gap solitons at the interface between uniform media and an optical lattice with defocusing nonlinearity. Such new type of solitons forms when the incident and reflected waves at the interface of the lattice experience Bragg scattering, and feature a combination of the unique properties of both surface waves and gap solitons. We discover that gap surface solitons exist only when the lattice depth exceeds a threshold value, that they can be made completely stable, and that they can form stable bound states.

Journal Article↗

PubMed2000

ICTS, an interventional cardiology training system.

In this article, we present an Interventional Cardiology Training System developed by the Medical Application Group at Mitsubishi Electric in collaboration with the Center for Innovative Minimally Invasive Therapy. The core of the ICTS is a computer simulation of interventional cardiology catheterization. This simulation integrates clinical expertise, research in learning, and technical innovations to create a realistic simulated environment. The goal of this training system is to augment the training of new cardiology fellows as well as to introduce cardiologists to new devices and procedures. To achieve this goal, both the technical components and the educational content of the ICTS bring new and unique features: a simulated fluoroscope, a physics model of a catheter, a haptic interface, a fluid flow simulation combined with a hemodynamic model and a learning system integrated in a user interface. The simulator is currently able to generate--in real-time--high quality x-ray images from a 3D anatomical model of the thorax, including a beating heart and animated lungs. The heart and lung motion is controlled by the hemodynamic model, which also computes blood pressure and EKG. The blood flow is then calculated according to the blood pressure and blood vessel characteristics. Any vascular tool, such as a catheter, guide wire or angioplasty balloon can be represented and accurately deformed by the flexible tool physics model. The haptics device controls the tool and provides appropriate feedback when contact with a vessel wall is detected. When the catheter is in place, a contrast agent can be injected into the coronary arteries; blood and contrast mixing is computed and a visual representation of the angiogram is displayed by the x-ray renderer. By bringing key advances in the area of medical simulation--with the real-time x-ray renderer for instance--and by integrating in a single system both high quality simulation and learning tools, the ICTS opens new perspectives for computer based training systems.

Angioplasty, Balloon, Coronary↗

PubMed1994

HLA-G polymorphisms in African Americans.

HLA-G is a class I MHC gene most notable for its restricted tissue distribution. The unique expression of this gene in extravillous cytotrophoblast at the maternal-fetal interface suggests that HLA-G plays a critical role in human pregnancy. Previous studies have reported that HLA-G has a limited repertoire of rare alleles, unlike the classical class I alleles, which are among the most polymorphic human genes. To determine the full extent of sequence variation in this gene, we examined the nucleotide variation in the first six exons of this gene in 45 healthy African American persons. By using sensitive techniques that detect > 95% of nucleotide substitutions, we detected two sequence variations in the alpha-1 domain (exon 2) and 24 sequence variations in the alpha-2 domain (exon 3) that result in amino acid substitutions. These data indicate that HLA-G is potentially capable of presenting a wide variety of peptides and of eliciting an allogeneic response, similar to the classical class I HLA genes.

Adult↗

PubMed2007

Neural engineering to produce in vitro nerve constructs and neurointerface.

OBJECTIVE: Recently, our laboratory recapitulated a natural form of axon growth that occurs between late embryogenesis and early adulthood. In this article, we describe how this novel neural engineering approach may be used to produce a nervous tissue interface to integrate disconnected motor and sensory functions for external control. METHODS: For nervous system repair, we recently developed a unique method to engineer nervous tissue constructs in vitro consisting of bundles of axons spanning two populations of neuronal somata. To integrate electronics and nervous tissue to transform electrophysiological signals into electronic signals, we have designed a nervous tissue interface. RESULTS: Our nervous tissue interface consists of stretch-grown nervous tissue with one end interfaced with a multiple electrode array, enabling us to detect and record real-time efferent signals conducted down the nerve and stimulate afferent sensory signaling. CONCLUSION: Our ultimate goal is to develop a neurally controlled prosthesis and a nervous system interface that could be linked to the patient's thoughts, providing two-way signaling for motor control and feedback from multiple external stimuli.

Axons↗

PubMed2003

Kinetic interaction analysis of human interleukin 5 receptor alpha mutants reveals a unique binding topology and charge distribution for cytokine recognition.

Human interleukin 5 receptor alpha (IL5Ralpha) comprises three fibronectin type III domains (D1, D2, and D3) in the extracellular region. Previous results have indicated that residues in the D1D2 domains are crucial for high affinity interaction with human interleukin 5 (IL5). Yet, it is the D2D3 domains that have sequence homology with the classic cytokine recognition motif that is generally assumed to be the minimum cytokine-recognizing unit. In the present study, we used kinetic interaction analysis of alanine-scanning mutational variants of IL5Ralpha to define the residues involved in IL5 recognition. Soluble forms of IL5Ralpha variants were expressed in S2 cells, selectively captured via their C-terminal V5 tag by anti-V5 tag antibody immobilized onto the sensor chip and examined for IL5 interaction by using a sandwich surface plasmon resonance biosensor method. Marked effects on the interaction kinetics were observed not only in D1 (Asp(55), Asp(56), and Glu(58)) and D2 (Lys(186) and Arg(188)) domains, but also in the D3 (Arg(297)) domain. Modeling of the tertiary structure of IL5Ralpha indicated that these binding residues fell into two clusters. The first cluster consists of D1 domain residues that form a negatively charged patch, whereas the second cluster consists of residues that form a positively charged patch at the interface of D2 and D3 domains. These results suggest that the IL5 x IL5Ralpha system adopts a unique binding topology, in which the cytokine is recognized by a D2D3 tandem domain combined with a D1 domain, to form an extended cytokine recognition interface.

Amino Acid Sequence↗

PubMed1996

Microbial lipases: production and applications.

Lipases occupy a prominent place among biocatalysts and have a wide spectrum of biotechnological applications. Lipases are unique as they hydrolyse fats into fatty acids and glycerol at the water-lipid interface and can reverse the reaction in non-aqueous media. The stability of these enzymes in organic solvents have pushed them into the frontier areas of organic synthesis leading to the designing of novel drugs, surfactants, bioactive compounds and oleochemicals. In addition, lipase-catalysed trans-esterification and inter-esterification reactions have been exploited in the fat industry. Looking into the wide scenario of lipase applications, commercialization of lipase production is a prime area of interest for microbiologists, process engineers and biochemists. Research carried out in this field has revealed that microbes, especially fungi and bacteria, are the tools of choice for commercial production. Recently, the structure determination of a few microbial lipases has widened our knowledge about the unique mechanism of catalysis of this enzyme.

Animals↗

PubMed2000

Cryptosporidium parvum induces host cell actin accumulation at the host-parasite interface.

Cryptosporidium parvum is an intracellular protozoan parasite that causes a severe diarrheal illness in humans and animals. Previous ultrastructural studies have shown that Cryptosporidium resides in a unique intracellular compartment in the apical region of the host cell. The mechanisms by which Cryptosporidium invades host intestinal epithelial cells and establishes this compartment are poorly understood. The parasite is separated from the host cell by a unique electron-dense structure of unknown composition. We have used indirect immunofluorescence microscopy and confocal laser scanning microscopy to characterize this structure. These studies indicate that host filamentous actin is assembled into a plaque-like structure at the host-parasite interface during parasite invasion and persists during parasite development. The actin-binding protein alpha-actinin is also present in this plaque early in parasite development but is lost as the parasite matures. Other actin-associated proteins, including vinculin, talin, and ezrin, are not present. We have found no evidence of tyrosine phosphorylation within this structure. Molecules known to link actin filaments to membrane were also examined, including alpha-catenin, beta-catenin, plakoglobin, and zyxin, but none was identified at the host-parasite junction. Thus, Cryptosporidium induces rearrangement of the host cell cytoskeleton and incorporates host cell actin and alpha-actinin into a host-parasite junctional complex.

Actins↗

PubMed2003

A review of drainage and spontaneous rupture in free standing thin films with tangentially immobile interfaces.

A review of spontaneous rupture in thin films with tangentially immobile interfaces is presented that emphasizes the theoretical developments of film drainage and corrugation growth through the linearization of lubrication theory in a cylindrical geometry. Spontaneous rupture occurs when corrugations from adjacent interfaces become unstable and grow to a critical thickness. A corrugated interface is composed of a number of waveforms and each waveform becomes unstable at a unique transition thickness. The onset of instability occurs at the maximum transition thickness, and it is shown that only upper and lower bounds of this thickness can be predicted from linear stability analysis. The upper bound is equivalent to the Frenkel criterion and is obtained from the zeroth order approximation of the H3 term in the evolution equation. This criterion is determined solely by the film radius, interfacial tension and Hamaker constant. The lower bound is obtained from the first order approximation of the H3 term in the evolution equation and is dependent on the film thinning velocity. A semi-empirical equation, referred to as the MTR equation, is obtained by combining the drainage theory of Manev et al. [J. Dispersion Sci. Technol., 18 (1997) 769] and the experimental measurements of Radoev et al. [J. Colloid Interface Sci. 95 (1983) 254] and is shown to provide accurate predictions of film thinning velocity near the critical thickness of rupture. The MTR equation permits the prediction of the lower bound of the maximum transition thickness based entirely on film radius, Plateau border radius, interfacial tension, temperature and Hamaker constant. The MTR equation extrapolates to Reynolds equation under conditions when the Plateau border pressure is small, which provides a lower bound for the maximum transition thickness that is equivalent to the criterion of Gumerman and Homsy [Chem. Eng. Commun. 2 (1975) 27]. The relative accuracy of either bound is thought to be dependent on the amplitude of the hydrodynamic corrugations, and a semi-empirical correlation is also obtained that permits the amplitude to be calculated as a function of the upper and lower bound of the maximum transition thickness. The relationship between the evolving theoretical developments is demonstrated by three film thickness master curves, which reduce to simple analytical expressions under limiting conditions when the drainage pressure drop is controlled by either the Plateau border capillary pressure or the van der Waals disjoining pressure. The master curves simplify solution of the various theoretical predictions enormously over the entire range of the linear approximation. Finally, it is shown that when the Frenkel criterion is used to assess film stability, recent studies reach conclusions that are contrary to the relevance of spontaneous rupture as a cell-opening mechanism in foams.

Journal Article↗

PubMed1990

Magnetic resonance imaging simulator: a teaching tool for radiology.

The increasing use of magnetic resonance imaging (MRI) as a clinical modality has put an enormous burden on medical institutions to cost effectively teach MRI scanning techniques to technologists and physicians. Since MRI scanner time is a scarce resource, it would be ideal if the teaching could be effectively performed off-line. In order to meet this goal, the radiology Department at the University of Pennsylvania has designed and developed a Magnetic Resonance Imaging Simulator. The simulator in its current implementation mimics the General Electric Signa (General Electric Magnetic Resonance Imaging System, Milwaukee, WI) scanner's user interface for image acquisition. The design is general enough to be applied to other MRI scanners. One unique feature of the simulator is its incorporation of an image-synthesis module that permits the user to derive images for any arbitrary combination of pulsing parameters for spin-echo, gradient-echo, and inversion recovery pulse sequences. These images are computed in 5 seconds. The development platform chosen is a standard Apple Macintosh II (Apple Computer, Inc, Cupertino, CA) computer with no specialized hardware peripherals. The user interface is implemented in HyperCard (Apple Computer Inc, Cupertino, CA). All other software development including synthesis and display functions are implemented under the Macintosh Programmer's Workshop 'C' environment. The scan parameters, demographics, and images are tracked using an Oracle (Oracle Corp, Redwood Shores, CA) data base. Images are currently stored on magnetic disk but could be stored on optical media with minimal effort.

Computer Simulation↗

PubMed1989

Ordered disruption of subunit interfaces during the stepwise reversible dissociation of Escherichia coli phosphofructokinase with KSCN.

The reversible inactivation and dissociation of the allosteric phosphofructokinase from Escherichia coli has been studied in relatively mild conditions, i.e., in the presence of the chaotropic agent KSCN. At moderate KSCN concentration, the loss of enzymatic activity involves two separated phases: first, a rapid dissociation of part of the tetramer into dimers, second, a slower displacement of the dimer-tetramer equilibrium upon further dissociation of the dimer into monomers. These two reactions can no longer be distinguished above 0.3 M KSCN since complete inactivation occurs in a single reaction. Different changes are observed for the fluorescence and the activity of the enzyme in KSCN: the fluorescence is not affected by the dissociation into dimers which is responsible for inactivation. The decrease in fluorescence reflects the change in environment of the unique tryptophan residue, Trp 311, during the dimer to monomer dissociation. This residue belongs to the interface containing the regulatory site, and its native fluorescence indicates that this interface is still present in the dimer. The substrate fructose 6-phosphate protects phosphofructokinase from inactivation by binding to the tetramer and prevents its dissociation into dimers. The presence of phosphoenolpyruvate prevents the slow dissociation of the dimer into monomers, which shows the ability of the dimer to bind the inhibitor. Two successive processes can be observed during reassociation of the protein upon KSCN dilution. First, a fast reaction (k1 = 2 x 10(5) M-1.s-1) is accompanied by a fluorescence increase and results in the formation of the dimeric species.(ABSTRACT TRUNCATED AT 250 WORDS)

Allosteric Regulation↗

PubMed2006

The crystal structure of human IRE1 luminal domain reveals a conserved dimerization interface required for activation of the unfolded protein response.

The unfolded protein response (UPR) is an evolutionarily conserved mechanism by which all eukaryotic cells adapt to the accumulation of unfolded proteins in the endoplasmic reticulum (ER). Inositol-requiring kinase 1 (IRE1) and PKR-related ER kinase (PERK) are two type I transmembrane ER-localized protein kinase receptors that signal the UPR through a process that involves homodimerization and autophosphorylation. To elucidate the molecular basis of the ER transmembrane signaling event, we determined the x-ray crystal structure of the luminal domain of human IRE1alpha. The monomer of the luminal domain comprises a unique fold of a triangular assembly of beta-sheet clusters. Structural analysis identified an extensive dimerization interface stabilized by hydrogen bonds and hydrophobic interactions. Dimerization creates an MHC-like groove at the interface. However, because this groove is too narrow for peptide binding and the purified luminal domain forms high-affinity dimers in vitro, peptide binding to this groove is not required for dimerization. Consistent with our structural observations, mutations that disrupt the dimerization interface produced IRE1alpha molecules that failed to either dimerize or activate the UPR upon ER stress. In addition, mutations in a structurally homologous region within PERK also prevented dimerization. Our structural, biochemical, and functional studies in vivo altogether demonstrate that IRE1 and PERK have conserved a common molecular interface necessary and sufficient for dimerization and UPR signaling.

Amino Acid Sequence↗

PubMed2002

Atmospheric pressure laser desorption/ionization on porous silicon.

A recently developed commercial atmospheric pressure matrix-assisted laser desorption/ionization (AP-MALDI) source (MassTech, Inc.) was modified to adopt commercially available DIOS plates (Mass Consortium Corp.) for the studies of laser desorption from the surface of porous silicon under atmospheric pressure conditions. The feasibility of atmospheric pressure laser desorption/ionization from the surface of porous silicon (AP-DIOS) was demonstrated. The advantages of this new AP-DIOS technique include reasonably good sensitivity (subpicomole range for standard peptide mixtures), simplicity of sample preparation, uniformity of target spots and the absence of matrix peaks in the spectra. The AP-DIOS source was interfaced with a commercial ion trap (LCQ Classic, Thermo Finnigan) which additionally provides a unique MS(n) capability. The AP-DIOS spectrum of 250 fmol of unseparated tryptic digest of bovine serum albumin (BSA) was compared with that of AP-MALDI for the same compound. AP-DIOS offers significantly better coverage for the digest components in the mass range 200-1000 Da. The combined data of both techniques enabled us to nearly double the number of matched peaks in BSA digest analysis compared with AP-DIOS or AP-MALDI analysis separately.

Angiotensins↗

PubMed1983

Submersion culture of the intact fetal lung.

Intact whole lungs from 14 d rat fetuses were explanted to organ culture and maintained fully submerged in medium or positioned at the gas-fluid interface. Striking differences were observed in the morphological development of submersion cultured and interface cultured explants after 7 d in vitro. Submersion cultured lung developed as an expanded hollow organ, whereas the tissue of interface cultures was glandular in appearance and attenuated against the culture substrate. This difference in the development of the cultured lung may be attributed to the influence of surface tension forces at the gas-fluid interface versus the fluid immersed environment. In submersion culture the fetal lung followed a unique pattern of morphogenetic development. Under these culture conditions the lung expanded in three dimensions and retained its discrete lobar structure. The primitive bronchial epithelial tree of the fetal lung at explantation exhibited substantial histoarchitectural changes in vitro. Progressive branching of the epithelium into the surrounding lung mesenchyme gave rise to a pulmonary parenchyma bearing characteristics of the presumptive alveolar region of the late gestation fetal lung. The epithelium consisted predominantly of large, glycogen rich cuboidal cells interspersed with numerous structurally differentiated type II cells, containing lamellar bodies. The pulmonary parenchyma of the submersion cultured explant occupied a narrow margin of the lung wall, surrounding a large central lumen. This unique form of the cultured lung made it possible to examine the pulmonary epithelium by scanning electron microscopy. Data from the present study suggest that submersion culture of the intact fetal lung may provide a useful model system for study of the maturation of epithelial cell surface features during pulmonary development.

Animals↗

PubMed2005

Influence of process parameters on the size distribution of PLA microcapsules prepared by combining membrane emulsification technique and double emulsion-solvent evaporation method.

Relatively uniform-sized biodegradable poly(lactide) (PLA) microcapsules with various sizes were successfully prepared by combining a glass membrane emulsification technique and water-in-oil-in-water (w1/o/w2) double emulsion-solvent evaporation method. A water phase was used as the internal water phase, a mixture solvent of dichloromethane (DCM) and toluene dissolving PLA and Arlacel 83 was used as the oil phase (o). These two solutions were emulsified by a homogenizer to form a w1/o primary emulsion. The primary emulsion was permeated through the uniform pores of a glass membrane into the external water phase by the pressure of nitrogen gas to form the uniform w1/o/w2 double emulsion droplets. Then, the solid polymer microcapsules were obtained by simply evaporating solvent. The influence of process parameters on the size distribution of PLA microcapsules was investigated, with an emphasis on the effect of oil-soluble emulsifier. A unique phenomenon was found that a large part of emulsifier could adsorb on the interface of internal water phase and oil phase, which suppressed its adsorption on the surface of glass membrane, and led to the successful preparation of uniform-sized double emulsion. Finally, by optimizing the process parameters, PLA microcapsules with various sizes having coefficient of variation (CV) value under 14.0% were obtained. Recombinant human insulin (rhI), as a model protein, was encapsulated into the microcapsules with difference sizes, and its encapsulation efficiency and cumulative release were investigated. The result suggested that the release behavior could be simply adjusted just by changing precisely the diameters of microcapsule, benefited from the membrane emulsification technique.

Capsules↗