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PubMed2004

The unique regulation of brain cytochrome P450 2 (CYP2) family enzymes by drugs and genetics.

Cytochrome P450 (CYP) enzymes in the brain may have a role in the activation or inactivation of centrally acting drugs, in the metabolism of endogenous compounds, and in the generation of damaging toxic metabolites and/or oxygen stress. CYPs are distributed unevenly among brain regions, and are found in neurons, glial cells and at the blood-brain interface. They have been observed in mitochondrial membranes, in neuronal processes and in the plasma membrane, as well as in endoplastic reticulum. Brain CYPs are inducible by many common hepatic inducers, however many compounds affect liver and brain CYP expression differently, and some CYPs which are constitutively expressed in liver are inducible in brain. CYP induction is isozyme-, brain region-, cell type- and inducer-specific. While it is unlikely that brain CYPs contribute to overall clearance of xenobiotics, their punctate, region- and cell-specific expression suggests that CNS CYPs may create micro-environments in the brain with differing drug and metabolite levels (not detected or predicted by plasma drug monitoring). Coupled with the sensitivity of CNS CYPs to induction, this may in part account for inter-individual variation in response to centrally acting drugs and neurotoxins, and may have implications for individual variation in receptor adaptation and cross-tolerance to different drugs. In addition, genetic variation in brain CYPs, depending on the type of polymorphism (structural versus regulatory), will alter enzyme activity. These aspects of brain CYP expression regulation and genetic influences are illustrated in this review using mRNA, protein, and enzyme activity data for CYP2D1/6, CYP2E1 and CYP2B1/6 in rat and human brain. The role of CYP-mediated metabolism in the brain, a highly heterogeneous and complex organ, is a new and relatively unexplored field of scientific enquiry. It holds promise for furthering our undestanding of inter-individual variability in response to centrally acting drugs as well as risk for neurological diseases and pathogies.

Animals↗

PubMed1984

Perceived variability and symbol use: a common language-cognition interface in children and chimpanzees (Pan troglodytes).

Analysis of two chimpanzees' conversations with their teacher during a tool-use training task demonstrated that chimps use lexigrams, a humanly devised visual symbol system, selectively to encode perceived variability; that is, they generally used their symbols to differentiate alternative possibilities or to represent change or novelty in a situation. In contrast, they tended to leave unsaid what was unchanging, repetitive, or the unique possibility in a situation. Perceived variability influenced not only which symbols were selected but also utterance length: A single dimension of variability in a situation leads to single-lexigram utterances; multiple dimensions are associated with multi-lexigram utterances. This pattern of results indicates that the absence of formal grammatical structure in chimp language does not imply that utterances beyond one word in length are either rote strings or imitations. The chimps' tendency to mention the variable while leaving the constant or redundant unsaid is, moreover, strong support for the position that their use of a humanly devised symbol system is more than a series of conditioned responses.

Animals↗

PubMed2000

A brain-controlled switch for asynchronous control applications.

Asynchronous control applications are an important class of application that has not received much attention from the brain-computer interface (BCI) community. This work provides a design for an asynchronous BCI switch and performs the first extensive evaluation of an asynchronous device in attentive, spontaneous electroencephalographic (EEG). The switch design [named the low-frequency asynchronous switch design (LF-ASD)] is based on a new feature set related to imaginary movements in the 1-4 Hz frequency range. This new feature set was identified from a unique analysis of EEG using a bi-scale wavelet. Offline evaluations of a prototype switch demonstrated hit (true positive) rates in the range of 38%-81% with corresponding false positive rates in the range of 0.3%-11.6%. The performance of the LF-ASD was contrasted with two other ASDs: one based on mu-power features and another based on the outlier processing method (OPM) algorithm. The minimum mean error rates for the LF-ASD were shown to be significantly lower than either of these other two switch designs.

Adult↗

PubMed1991

Structural analysis of the peptidyl transferase region in ribosomal RNA of the eukaryote Xenopus laevis.

Accessible single-strand bases in Xenopus laevis 28 S ribosomal RNA (rRNA) Domain V, the peptidyl transferase region, were determined by chemical modification with dimethylsulfate, 1-cyclohexyl-3-(2-morpholinoethyl-carbodiimide metho-p-toluene sulfonate and kethoxal, followed by primer extension. The relative accessibilities of three rRNA substrates were compared: deproteinized 28 S rRNA under non-denaturing conditions (free 28 S rRNA), 60 S subunits and 80 S ribosomes. Overall, our experimental results support the theoretical secondary structure model of Domain V derived by comparative sequence analysis and compensatory base-pair changes, and support some theoretical tertiary interactions previously suggested by covariation. The 60 S subunits and 80 S ribosomes generally show increasing resistance to chemical modification. Bases which are sensitive in free 28 S rRNA but protected in 60 S subunits may be sites for ribosomal protein binding or induced structural rearrangements. Another class of nucleotides is distinguished by its sensitivity in 60 S subunits but protection in 80 S ribosomes; these nucleotides may be involved in subunit-subunit interactions or located at the interface of the ribosome. We found a third class of bases, which is protected in free 28 S rRNA but sensitive in 60 S subunits and/or 80 S ribosomes, suggesting that structural changes occur in Domain V as a result of subunit assembly and ribosome formation. One such region is uniquely hypersensitive in eukaryotic ribosomes but is absent in Escherichia coli ribosomes. Sites that we determined to be accessible on empty 80 S ribosomes could serve as recognition sites for translation components.

Aldehydes↗

PubMed2002

Solution NMR structure of S100B bound to the high-affinity target peptide TRTK-12.

The solution NMR structure is reported for Ca(2+)-loaded S100B bound to a 12-residue peptide, TRTK-12, from the actin capping protein CapZ (alpha1 or alpha2 subunit, residues 265-276: TRTKIDWNKILS). This peptide was discovered by Dimlich and co-workers by screening a bacteriophage random peptide display library, and it matches exactly the consensus S100B binding sequence ((K/R)(L/I)XWXXIL). As with other S100B target proteins, a calcium-dependent conformational change in S100B is required for TRTK-12 binding. The TRTK-12 peptide is an amphipathic helix (residues W7 to S12) in the S100B-TRTK complex, and helix 4 of S100B is extended by three or four residues upon peptide binding. However, helical TRTK-12 in the S100B-peptide complex is uniquely oriented when compared to the three-dimensional structures of other S100-peptide complexes. The three-dimensional structure of the S100B-TRTK peptide complex illustrates that residues in the S100B binding consensus sequence (K4, I5, W7, I10, L11) are all involved in the S100B-peptide interface, which can explain its orientation in the S100B binding pocket and its relatively high binding affinity. A comparison of the S100B-TRTK peptide structure to the structures of apo- and Ca(2+)-bound S100B illustrates that the binding site of TRTK-12 is buried in apo-S100B, but is exposed in Ca(2+)-bound S100B as necessary to bind the TRTK-12 peptide.

Amino Acid Sequence↗

PubMed1998

Structure of the dimer initiation complex of HIV-1 genomic RNA.

Retroviral genomes must dimerize to be fully infectious. Dimerization is directed by a unique RNA hairpin structure with a palindrome in its loop: hairpins of two strands first associate transiently through their loops, and then refold to a more stable, linear duplex. The structure of the initial, kissing-loop dimer from HIV-1, solved using 2D NMR, is bent and metastable, its interface being formed not only by standard basepairing between palindromes, but also by a distinctive pattern of interstrand stacking among bases at the stem-loop junctions. This creates mechanical distortions that partially melt both stems, which may facilitate spontaneous refolding of this RNA complex into linear form.

Base Composition↗

PubMed1996

Domain swapping creates a third putative combining site in bovine odorant binding protein dimer.

In mammals, odorant binding proteins may play an important role in the transport of odors towards specific olfactory receptors on sensory neurones across the aqueous compartment of the nasal mucus. We have solved the X-ray structure of such a transport protein, bovine odorant binding protein (OBP) at 2.0 A resolution. The beta-barrel of OBP is similar to that of lipocalins, but OBP dimer association results from domain swapping, an observation unique among the lipocalins. The alpha-helix of each monomer stacks against the beta-barrel of the other monomer. Contrary to previous reports, each monomer has an internal buried cavity which could accommodate a naturally occurring molecule. Besides this cavity, an open cavity is located at the dimer interface. Data in solution suggest that this central cavity may be a binding site created by domain swapping.

Animals↗

PubMed2000

Crystal structure of alkaline phosphatase from human placenta at 1.8 A resolution. Implication for a substrate specificity.

Human placental alkaline phosphatase (PLAP) is one of three tissue-specific human APs extensively studied because of its ectopic expression in tumors. The crystal structure, determined at 1.8-A resolution, reveals that during evolution, only the overall features of the enzyme have been conserved with respect to Escherichia coli. The surface is deeply mutated with 8% residues in common, and in the active site, only residues strictly necessary to perform the catalysis have been preserved. Additional structural elements aid an understanding of the allosteric property that is specific for the mammalian enzyme (Hoylaerts, M. F., Manes, T., and Millán, J. L. (1997) J. Biol. Chem. 272, 22781-22787). Allostery is probably favored by the quality of the dimer interface, by a long N-terminal alpha-helix from one monomer that embraces the other one, and similarly by the exchange of a residue from one monomer in the active site of the other. In the neighborhood of the catalytic serine, the orientation of Glu-429, a residue unique to PLAP, and the presence of a hydrophobic pocket close to the phosphate product, account for the specific uncompetitive inhibition of PLAP by l-amino acids, consistent with the acquisition of substrate specificity. The location of the active site at the bottom of a large valley flanked by an interfacial crown-shaped domain and a domain containing an extra metal ion on the other side suggest that the substrate of PLAP could be a specific phosphorylated protein.

Alkaline Phosphatase↗

PubMed2002

Conspicuous veils formed by vibrioid bacteria on sulfidic marine sediment.

We describe the morphology and behavior of a hitherto unknown bacterial species that forms conspicuous veils (typical dimensions, 30 by 30 mm) on sulfidic marine sediment. The new bacteria were enriched on complex sulfidic medium within a benthic gradient chamber in oxygen-sulfide countergradients, but the bacteria have so far not been isolated in pure culture, and a detailed characterization of their metabolism is still lacking. The bacteria are colorless, gram-negative, and vibrioid-shaped (1.3- to 2.5- by 4- to 10- micro m) cells that multiply by binary division and contain several spherical inclusions of poly-beta-hydroxybutyric acid. The cells have bipolar polytrichous flagella and exhibit a unique swimming pattern, rotating and translating along their short axis. Free-swimming cells showed aerotaxis and aggregated at ca. 2 micro M oxygen within opposing oxygen-sulfide gradients, where they were able to attach via a mucous stalk, forming a cohesive whitish veil at the oxic-anoxic interface. Bacteria attached to the veil kept rotating and adapted their stalk lengths dynamically to changing oxygen concentrations. The joint action of rotating bacteria on the veil induced a homogeneous water flow from the oxic water region toward the veil, whereby the oxygen uptake rate could be enhanced up to six times, as shown by model calculations. The veils showed a pronounced succession pattern. New veils were generated de novo within 24 h and had a homogeneous whitish translucent appearance. Bacterial competitors or eukaryotic predators were apparently kept away by the low oxygen concentration prevailing at the veil surface. Frequently, within 2 days the veil developed a honeycomb pattern of regularly spaced holes. After 4 days, most veils were colonized by grazing ciliates, leading to the fast disappearance of the new bacteria. Several-week-old veils finally developed into microbial mats consisting of green, purple, and colorless sulfur bacteria.

Bacteria↗

PubMed2001

Manipulation of volumetric patient data in a distributed virtual reality environment.

Due to increases in network speed and bandwidth, distributed exploration of medical data in immersive Virtual Reality (VR) environments is becoming increasingly feasible. The volumetric display of radiological data in such environments presents a unique set of challenges. The shear size and complexity of the datasets involved not only make them difficult to transmit to remote sites, but these datasets also require extensive user interaction in order to make them understandable to the investigator and manageable to the rendering hardware. A sophisticated VR user interface is required in order for the clinician to focus on the aspects of the data that will provide educational and/or diagnostic insight. We will describe a software system of data acquisition, data display, Tele-Immersion, and data manipulation that supports interactive, collaborative investigation of large radiological datasets. The hardware required in this strategy is still at the high-end of the graphics workstation market. Future software ports to Linux and NT, along with the rapid development of PC graphics cards, open the possibility for later work with Linux or NT PCs and PC clusters.

Computer Communication Networks↗

PubMed1998

Backside nonconformity and locking restraints affect liner/shell load transfer mechanisms and relative motion in modular acetabular components for total hip replacement.

Nonconformity between the polyethylene liner and the metal shell may exist in modular acetabular components by design, due to manufacturing tolerances, or from locking mechanisms that attach the polyethylene liner to the metal shell. Relative motion at the liner/shell interface has been associated with backside wear, which may contribute to osteolysis which has been clinically observed near screw holes. The purpose of this study was to investigate the effect of nonconformity and locking restraints on the liner/shell relative motion and load transfer mechanisms in a commercially available, metal-backed acetabular component with a polar fenestration. The finite element method was used to explore the hypothesis that backside nonconformity and locking restraints play important roles in long-term surface damage mechanisms that are unique to modular components, such as backside wear and liner extrusion through screw holes. The three-body quasi-static contact problem was solved using a commercially available explicit finite element code, which modeled contact between the femoral head, polyethylene liner, and the metal shell. Four sets of liner boundary conditions were investigated: no restraints, rim restraints, equatorial restraints, and both rim and equatorial restraints. The finite element model with a conforming shell predicted between 8.5 and 12.8 microm of incremental extrusion of the polyethylene through the polar fenestration, consistent with in vitro experiments of the same design under identical loading conditions. Furthermore, idealized rim and/or equatorial liner restraints were found to share up to 71% of the load across the liner/shell interface. Consequently, the results of this study demonstrate that backside nonconformity and locking restraints substantially influence backside relative motion as well as load transfer at the liner/shell interface.

Acetabulum↗

PubMed1997

Glial-glial and glial-neuronal interfaces in radiation-induced, glia-depleted spinal cord.

This review summarises some of the major findings derived from studies using the model of a glia-depleted environment developed and characterised in this laboratory. Glial depletion is achieved by exposure of the immature rodent spinal cord to x-radiation which markedly reduces both astrocyte and oligodendrocyte populations and severely impairs myelination. This glia-depleted, hypomyelinated state presents a unique opportunity to examine aspects of spinal cord maturation in the absence of a normal glial population. An associated sequela within 2-3 wk following irradiation is the appearance of Schwann cells in the dorsal portion of the spinal cord. Characteristics of these intraspinal Schwann cells, their patterns of myelination or ensheathment, and their interrelations with the few remaining central glia have been examined. A later sequela is the development of Schwann cells in the ventral aspect of the spinal cord where they occur predominantly in the grey matter. Characteristics of these ventrally situated intraspinal Schwann cells are compared with those of Schwann cells located dorsally. Recently, injury responses have been defined in the glia-depleted spinal cord subsequent to the lesioning of dorsal spinal nerve roots. In otherwise normal animals, dorsal nerve root injury induces an astrocytic reaction within the spinal segments with which the root(s) is/are associated. Lesioning of the 4th lumbar dorsal root on the right side in irradiated or nonirradiated animals results in markedly different glial responses with little astrocytic scarring in the irradiated animals. Tracing studies reveal that these lesioned dorsal root axons regrow rather robustly into the spinal cord in irradiated but not in nonirradiated animals. To examine role(s) of glial cells in preventing this axonal regrowth, glial cells are now being added back to this glia-depleted environment through transplantation of cultured glia into the irradiated area. Transplanted astrocytes establish barrier-like arrangements within the irradiated cords and prevent axonal regrowth into the cord. Studies using other types of glial cultures (oligodendrocyte or mixed) are ongoing.

Animals↗

PubMed1989

Standardized acquisition of bedside data: the IEEE P1073 medical information bus.

The absence of standards for medical device communications has stymied the acceptance and success of automated clinical data management systems. Even devices with simple RS-232 data output ports require special interfacing hardware and software. Due to the number and variety of medical devices available, each with their own peculiar data output configuration, it has been impractical to interface with most of them. Limited by manual data entry, most computerized patient data management systems have failed to deliver the productivity gains their users expected. The forthcoming IEEE P1073 Medical Information Bus (MIB) Standard promises to correct this situation with a single powerful bedside device interface method. The MIB will provide specifications for all hardware and software necessary for medical data communications. The MIB handles the need for automatic recognition of new devices placed at a bedside, automatic reconfiguration of the network, binding of a device to a particular patient's bedside and many other issues unique to the medical data communications environment. The MIB is expected to undergo formal IEEE balloting in 1990 and promises to open a new era in data management for clinical patient care.

Computer Systems↗

PubMed2004

Differential down-modulation of HLA-G and HLA-A2 or -A3 cell surface expression following human cytomegalovirus infection.

During pregnancy, the non-classical major histocompatibility complex (MHC) class I HLA-G molecule is specifically expressed in trophoblast cells at the materno-fetal interface and may exert a local control of the immune response against viral infections. Human cytomegalovirus (HCMV) infection, which is the major cause of congenital defects, encodes multiple glycoproteins (US2, US3, US6, US10 and US11) that interrupt the MHC class I pathway of antigen presentation. The effect of some of these unique short (US) proteins on HLA-G expression has been previously studied, but little is known about the modulation of HLA-G cell surface expression during the course of HCMV infection which ensures expression of all of these US proteins. Using flow cytometry analysis, HLA-G cell surface expression was evaluated in HCMV-infected U373-HLA-G transfectant cells and compared with the modulation of the endogenous classical HLA-A2 molecules. The results indicated that HCMV infection down-modulated HLA-G cell surface expression, but later after infection and to a lesser extent than HLA-A2. Using various HLA-G/HLA-A2 chimeras, we showed that the unique structure of HLA-G cytoplasmic tail was partly involved in the resistance of HLA-G to viral down-modulation. Such limited down-modulation of HLA-G may have functional consequences in term of innate immunity against congenital HCMV infection.

Amino Acid Sequence↗

PubMed1998

The contribution of lipids and lipid metabolism to cellular functions of the Golgi complex.

The history of the Golgi complex now reaches its 100 year anniversary. Over the past several decades, tremendous effort has gone into cataloguing Golgi resident proteins, measuring the lipid compositions of Golgi membranes, and in elucidating the pathways by which proteins and lipids traffic through this unique organelle. Only in the past 8 years or so has experimental scrutiny extended to the investigation of how lipids and proteins cooperate to endow the Golgi with its various capabilities regarding protein/lipid transport and sorting. In this chapter we review some of the most recent advances in deciphering the functional interfaces between lipids and proteins of the Golgi complex.

Animals↗

PubMed1999

External skeletal fixation. Linear fixators.

Linear external skeletal fixators offer some unique advantages over other fixation systems; they are biomechanically versatile, minimize surgical trauma to the fracture environment, and allow for simple staged disassembly to help promote bone healing. Historically, the use of linear external skeletal fixators has been reported to result in numerous complications, primarily related to the pin-bone interface. External skeletal fixators have experienced a resurgence in the past two decades. Increased knowledge of proper pin insertion techniques and advancements in pin design have greatly enhanced the longevity of the pin-bone interface, resulting in fewer complications. This article reviews some of the potential advantages of linear external skeletal fixators and principles of application to help strengthen the pin-bone interface and minimize complications.

Animals↗

PubMed1997

Use of a polybrene capillary coating in capillary electrophoresis for rapid analysis of hemoglobin variants with on-line detection via an ion trap storage/reflectron time-of-flight mass spectrometer.

A polybrene capillary coating in capillary electrophoresis (CE) has been used for rapid analysis of hemoglobin variant digests. The use of the polybrene capillary coating has allowed sufficient separation to resolve the large number of digest products formed upon tryptic digestion of the whole protein, so that prior separation of the hemoglobin alpha and beta chains is not required. The resolution of the digest peaks obtained by CE is sufficient so that even single amino acid substitutions can easily be detected using UV absorption detection. The digest is further analyzed by capillary electrophoresis separation with on-line detection using electrospray ionization interfaced to the ion trap storage/reflectron time of flight device (CE/ESI-IT/reTOF), where a comparison of the total ion electropherograms and mass spectra of the mutant and normal hemoglobins can detect the presence of a mutation site. The CE separation and mass analysis can be accomplished in typically 10-15 min. The unique capability of the CE/ESI-IT/reTOF system for detection of fast separations with narrow peaks that may be under 1 s fwhm is demonstrated. The speed of this system is essential for resolution of the large number of peaks that are separated in a short time duration using CE separations.

Adult↗

PubMed2005

Coupling cell proliferation and development in plants.

Plant genome projects have revealed that both the cell-cycle components and the overall cell-cycle architecture are highly evolutionarily conserved. In addition to the temporal and spatial regulation of cell-cycle progression in individual cells, multicellularity has imposed extra layers of complexity that impinge on the balance of cell proliferation and growth, differentiation and organogenesis. In contrast to animals, organogenesis in plants is a postembryonic and continuous process. Differentiated plant cells can revert to a pluripotent state, proliferate and transdifferentiate. This unique potential is strikingly illustrated by the ability of certain cells to produce a mass of undifferentiated cells or a fully totipotent embryo, which can regenerate mature plants. Conversely, plant cells are highly resistant to oncogenic transformation. This review discusses the role that cell-cycle regulators may have at the interface between cell division and differentiation, and in the context of the high plasticity of plant cells.

Cell Cycle Proteins↗