Pulse transit time: a new approach to haemodynamic monitoring in obstetric spinal anaesthesia

Part of the Portfolio Thesis by Geoffrey H. Sharwood-Smith: The inferior vena caval compression theory of hypotension in obstetric spinal anaesthesia: studies in normal and preeclamptic pregnancy, a literature review and revision of fundamental concepts, available at http://hdl.handle.net/10023/1815Original abstract presented at the Obstetric Anaesthetisits' Association congress 2002, Nottingham, 9-10 May.Postprin

Pulse transit time confirms altered response to spinal anaesthesia in pregnancy induced hypertension

Poster presented at the International Society for the Study of Hypertension in Pregnancy (ISSHP)Congress, Toronto 2002.Part of the Portfolio Thesis by Geoffrey H. Sharwood-Smith: The inferior vena caval compression theory of hypotension in obstetric spinal anaesthesia: studies in normal and preeclamptic pregnancy, a literature review and revision of fundamental concepts, available at http://hdl.handle.net/10023/1815Postprin

Analysis of polyubiquitin conjugates reveals that the Rpn10 substrate receptor contributes to the turnover of multiple proteasome targets

The polyubiquitin receptor Rpn10 targets ubiquitylated Sic1 to the 26S proteasome for degradation. In contrast, turnover of at least one ubiquitin-proteasome system (UPS) substrate, CPY*, is impervious to deletion of RPN10. To distinguish whether RPN10 is involved in the turnover of only a small set of cell cycle regulators that includes Sic1 or plays a more general role in the UPS, we sought to develop a general method that would allow us to survey the spectrum of ubiquitylated proteins that selectively accumulate in rpn10 cells. Polyubiquitin conjugates from yeast cells that express hexahistidine-tagged ubiquitin (H6-ubiquitin) were first enriched on a polyubiquitin binding protein affinity resin. This material was then denatured and subjected to IMAC to retrieve H6-ubiquitin and proteins to which it may be covalently linked. Using this approach, we identified 127 proteins that are candidate substrates for the 26S proteasome. We then sequenced ubiquitin conjugates from cells lacking Rpn10 (rpn10) and identified 54 proteins that were uniquely recovered from rpn10 cells. These include two known targets of the UPS, the cell cycle regulator Sic1 and the transcriptional activator Gcn4. Our approach of comparing the ubiquitin conjugate proteome in wild-type and mutant cells has the resolving power to identify even an extremely inabundant transcriptional regulatory protein and should be generally applicable to mapping enzyme substrate networks in the UPS

Anisotropic focusing characteristics of micro-domain structures within crystalline Sr_0.61Ba_0.39Nb₂O₆ : the crystal ball

We report the anisotropic focusing characteristics of a spherically configured region of micro-domains that have been induced within a cubic shaped crystal of Ce:doped Sr0.61Ba0.39Nb2O6. The internal spherical structure focuses extraordinary polarised light, but not ordinary polarised. The spherical region, which is easily observed via scattering, is formed as the crystal cools down, after a repoling cycle through the Curie temperature, with an applied field. Analytic modelling of the thermal gradients that exist within the crystal during cooling reveals a small (< 1°) temperature difference between the central and outside regions. The similarity in shape between these temperature profiles and the observed scattering region suggests a possible mechanism for the growth of this spherical micro-domained structure

The steady-state repertoire of human SCF Ubiquitin ligase complexes does not require ongoing Nedd8 conjugation

The human genome encodes 69 different F-box proteins (FBPs), each of which can potentially assemble with Skp1-Cul1-RING to serve as the substrate specificity subunit of an SCF ubiquitin ligase complex. SCF activity is switched on by conjugation of the ubiquitin- like protein Nedd8 to Cul1. Cycles of Nedd8 conjugation and deconjugation acting in conjunction with the Cul1-sequestering factor Cand1 are thought to control dynamic cycles of SCF assembly and disassembly, which would enable a dynamic equilibrium between the Cul1- RING catalytic core of SCF and the cellular repertoire of FBPs. To test this hypothesis, we determined the cellular composition of SCF complexes and evaluated the impact of Nedd8 conjugation on this steady-state. At least 42 FBPs assembled with Cul1 in HEK 293 cells, and the levels of Cul1-bound FBPs varied by over two orders of magnitude. Unexpectedly, quantitative mass spectrometry revealed that blockade of Nedd8 conjugation led to a modest increase, rather than a decrease, in the overall level of most SCF complexes. We suggest that multiple mechanisms including FBP dissociation and turnover cooperate to maintain the cellular pool of SCF ubiquitin ligases

The effect of signal noise on the remote sensing of Foliar biochemical concentration

Spectral measurements made using an imaging spectrometer contain systematic and random noise, while the former can be corrected the latter remains a source of error in the remotely sensed signal. A number of investigators have tried to determine the signal-to-noise-ratio (SNR) of the instrument, or the resultant imagery. However, the level of noise at which spectra are too noisy to be useful is not usually determined. The first attempt was by Goetz and Calvin, who suggested that the depth of the absorption feature should be at least an order of magnitude greater than the noise and more recently Dekker suggested a SNR of around 600:1 was required in visible/near infrared wavelengths to measure a 1/gl change in chlorophyll a concentration water. The wide range of applications of imaging spectroscopy make it difficult to set SNR specifications as they are dependent on a number of factors, one of the most important being reflectance of a particular target. For example, the SNR of imagery for vegetated targets is relatively low simply because vegetation has a relatively low level of reflectance. The Airborne Visible/Infrared Imaging Spectrometer (AVIRIS) is being used to estimate the concentration of biochemicals within vegetation canopies. This paper reports a study undertaken to identify first, wavebands that were highly correlated with foliar biochemical concentration and second, to determine how sensitive these correlations were to sensor noise

O-linked glycosylation sites profiling in Mycobacterium tuberculosis culture filtrate proteins

Mycobacterium tuberculosis (Mtb) causes tuberculosis, one of the leading causes of fatal infectious diseases worldwide. Cell–cell recognition between the pathogen Mtb and its host is mediated in part by glycosylated proteins. So far, glycoproteins in Mtb are understudied and for only very few glycoproteins glycosylation sites have been described, e.g., alanine and proline rich secreted protein apa, superoxide dismutase SODC, lipoprotein lpqH and MPB83/MPT83. In this study, glycosylated proteins in Mtb culture filtrate were investigated using liquid chromatography–mass spectrometry approaches and bioinformatic analyses. To validate the presence of glycoproteins, several strategies were pursued including collision induced dissociation, high energy collision dissociation and electron transfer dissociation techniques, and bioinformatics analyses involving a neutral loss search for glycosylated moieties. After extensive data curation, we report glycosylation sites for thirteen Mtb glycoproteins using a combination of mass spectrometry techniques on a dataset collected from culture filtrate proteins. This is the first glycoproteomics study identifying glycosylation sites on mycobacterial culture filtrate proteins (CFP) on a global scale. Biological significance In this study, glycosylation sites in Mtb were characterized by collision-induced dissociation, electron-transfer dissociation and high energy collision dissociation techniques. The identification of glycosylation sites is important for our understanding of the physiology and pathophysiology of Mtb. Glycoproteins are often responsible for protein–protein interactions between host and pathogen and thus represent interesting targets for vaccine development. In addition, our strategy is not limited to Mtb, but could be extended to other organisms

Exploring the remote sensing of foliar biochemical concentrations with AVIRIS data

Airborne Visible/Infrared Imaging Spectrometer (AVIRIS) data shows promise for the estimation of foliar biochemical concentrations at the scale of the canopy. There are, however, several problems associated with the use of AVIRIS data in this way and these are detailed in recent Plant Biochemical Workshop Report. The research reported was concentrated upon three of these problems: field sampling of forest canopies, wet laboratory assay of foliar chemicals, and the visualization of AVIRIS data