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1.
Anaesth Intensive Care ; 41(2): 184-201, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23530785

ABSTRACT

Interventional neuroradiology is a rapidly expanding field, and the complexity and duration of these procedures makes anaesthetic support essential to their success. Such has been the development in this area, that the American Heart Association has published a scientific statement on the indications for these procedures. A detailed understanding of patient pathology, the technical aspects of the interventions and their associated risks, and the remote location in which they are performed are important for providing expert anaesthetic care. The aim of this article is to provide a description and contemporary analysis of the common interventional neuroradiology procedures relevant to the anaesthetist. This article will cover the management of intracranial aneurysms, cerebral vasospasm following intracranial haemorrhage, intracranial and spinal arteriovenous malformations, idiopathic intracranial hypertension, carotid artery stenting, intra-arterial thrombolysis for stroke and endovascular treatment of intracranial atherosclerosis. Protection from ionising radiation and acute kidney injury are also discussed.


Subject(s)
Anesthesia/methods , Radiography, Interventional/methods , Embolization, Therapeutic , Humans , Intracranial Aneurysm/therapy , Intracranial Arteriovenous Malformations , Monitoring, Physiologic , Pseudotumor Cerebri/therapy , Stents , Stroke/therapy , Vasospasm, Intracranial/therapy
2.
J Immunol Methods ; 315(1-2): 19-26, 2006 Aug 31.
Article in English | MEDLINE | ID: mdl-16893550

ABSTRACT

Early stage rheumatoid arthritis (RA) is often difficult to diagnose because initial symptoms are non-specific. To aid diagnosis, suitable serological diagnostic markers are sought. Elevated levels of soluble MHC class II (soluble human leukocyte antigen; sHLA-DR) in human serum have been associated with rheumatoid and 'rheumatoid-like' autoimmune diseases. As a result, sHLA-DR has been suggested as a specific marker of RA. However, reported levels of sHLA-DR in sera of healthy donors vary significantly and the mechanism of release of HLA-DR into serum is poorly understood. Investigations into the diagnostic potential of this molecule necessitate the development of a sensitive and specific sHLA-DR assay. We have investigated multiple ELISA setups to develop such an assay and false positive signal has been carefully removed using a combination of isotype-matched controls and immuno-depletion. sHLA-DR levels in sera of RA patients were not significantly different from those in healthy donors which suggests sHLA-DR has limited utility in the diagnosis of RA. In RA patients, we detected high levels of sHLA-DR in aspirated synovial fluid (SF), but this did not correlate with sHLA-DR levels in serum.


Subject(s)
Arthritis, Rheumatoid/diagnosis , Enzyme-Linked Immunosorbent Assay/methods , HLA-DR Antigens/analysis , Synovial Fluid/metabolism , Arthritis, Rheumatoid/blood , Arthritis, Rheumatoid/immunology , Biomarkers/analysis , Case-Control Studies , Cell Line, Tumor , Female , HLA-DR Antigens/blood , Humans , Male , Middle Aged , Sensitivity and Specificity , Solubility , Synovial Fluid/immunology
3.
Stem Cells Dev ; 15(2): 221-31, 2006 Apr.
Article in English | MEDLINE | ID: mdl-16646668

ABSTRACT

Following the differentiation of cultured stem cells is often reliant on the expression of genes and proteins that provide information on the developmental status of the cell or culture system. There are few molecules, however, that show definitive expression exclusively in a specific cell type. Moreover, the reliance on a small number of molecules that are not entirely accurate biomarkers of particular tissues can lead to misinterpretation in the characterization of the direction of cell differentiation. Here we describe the use of technology that examines the mass spectrum of proteins expressed in cultured cells as a means to identify the developmental status of stem cells and their derivatives in vitro. This approach is rapid and reproducible and it examines the expression of several different biomarkers simultaneously, providing a profile of protein expression that more accurately corresponds to a particular type of cell differentiation.


Subject(s)
Cell Differentiation , Pluripotent Stem Cells/chemistry , Proteome/analysis , Proteomics/methods , Acetamides/pharmacology , Antigens, Surface/analysis , Antigens, Tumor-Associated, Carbohydrate , Biomarkers/analysis , Carcinoma, Embryonal/metabolism , Carcinoma, Embryonal/pathology , Embryonal Carcinoma Stem Cells , Flow Cytometry , Gangliosides/analysis , Glycosphingolipids/analysis , Humans , Keratins/analysis , Neoplastic Stem Cells , Neurons/chemistry , Neurons/pathology , Peptides/analysis , Pluripotent Stem Cells/drug effects , Pluripotent Stem Cells/pathology , Proteoglycans/analysis , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Stage-Specific Embryonic Antigens , Tretinoin/pharmacology , Tubulin/analysis
4.
J Biochem Biophys Methods ; 62(3): 231-40, 2005 Mar 31.
Article in English | MEDLINE | ID: mdl-15733583

ABSTRACT

Understanding neural differentiation and the development of complex neurite networks in three-dimensional matrices is critical for neural tissue engineering in vitro. In this study we describe for the first time the growth of human stem cell-derived neurons on solid polystyrene matrices coated with bioactive molecules. Highly porous foams were prepared from poly(styrene/divinylbenzene) using a high internal phase emulsion (HIPE) as a template to create the porous structure. The resulting polyHIPE matrices were readily coated with aqueous-based solutions including poly-d-lysine and laminin. Human neurons adhered well to poly-d-lysine coated surfaces and extended neural processes, however, neurite outgrowth was particularly enhanced when polymers also received a coating of laminin. These data clearly demonstrate the potential use of solid polystyrene scaffolds to create three-dimensional environments for cell growth and differentiation. We propose that these robust and stable matrices can be conveniently and routinely used in the tissue culture laboratory to study the behaviour of cells grown in three-dimensions.


Subject(s)
Cell Culture Techniques/instrumentation , Cell Culture Techniques/methods , Cell Differentiation , Neurons/cytology , Polymers/chemistry , Polymers/pharmacology , Stem Cells/cytology , Cell Line , Cell Proliferation/drug effects , Emulsions , Humans , Laminin/pharmacology , Microscopy, Electron, Scanning , Neurons/drug effects
5.
Stem Cells Dev ; 13(4): 400-8, 2004 Aug.
Article in English | MEDLINE | ID: mdl-15345134

ABSTRACT

There are few reliable experimental systems available to study the molecular mechanisms that govern human embryonic development. Embryonal carcinoma (EC) cells are pluripotent stem cells derived from teratocarcinomas and are considered the malignant counterparts of human embryonic stem (ES) cells. Several of the existing human EC stem cell lines provide robust and simple culture systems to study certain aspects of cellular differentiation in a manner pertinent to human embryogenesis. Here we review the strategies used to derive and characterize the established and recognized human EC stem cell line TERA2.cl.SP12. Furthermore, we demonstrate the value of human EC stem cells as a model of early development and focus on cell fate determination in the embryonic ectoderm.


Subject(s)
Carcinoma, Embryonal/pathology , Embryonic Development/physiology , Stem Cells/pathology , Cell Culture Techniques/methods , Cell Differentiation/physiology , Cell Line , Humans , Teratoma/pathology
6.
Biochem Biophys Res Commun ; 316(3): 918-23, 2004 Apr 09.
Article in English | MEDLINE | ID: mdl-15033489

ABSTRACT

The ability to effectively monitor the behaviour of pluripotent stem cells and their differentiation is key to their use in basic and clinical research. Molecules expressed in particular cell types can be used to report the status of cell differentiation and is a recognised means of assessing the behaviour of cell cultures. There are currently few useful markers of stem cells and there is no rapid way to accurately determine their level of expression. In this study, we describe for the first time the potential of surface enhanced laser desorption/ionisation time-of-flight mass spectrometry (SELDI-TOF-MS) to identify novel biomarkers of human pluripotent embryonal carcinoma stem cells and their differentiated derivatives. This approach allows the rapid and sensitive screening of cell samples without the need to purify the specimen prior to analysis. The identification of proteins expressed in specific cell populations will provide valuable tools for monitoring cellular development.


Subject(s)
Biomarkers , Proteomics/methods , Stem Cells/metabolism , Carcinoma, Embryonal/metabolism , Cell Differentiation , Cell Membrane/metabolism , Chromatography, Ion Exchange , Flow Cytometry , Fluorescent Antibody Technique, Indirect , Humans , Hydrogen-Ion Concentration , Mass Spectrometry , Microscopy, Phase-Contrast , Protein Array Analysis , Proteome
7.
Biochem Biophys Res Commun ; 314(2): 483-8, 2004 Feb 06.
Article in English | MEDLINE | ID: mdl-14733931

ABSTRACT

Growing and differentiating human stem cells in vitro can provide access to study the molecular mechanisms that control cellular development in a manner pertinent to human embryogenesis. To fully understand such processes, however, it is important to recreate culture conditions that most closely relate to those in living tissues. As step in this direction, we have developed a robust three-dimensional cell culture system using inert highly porous solid matrices manufactured from polystyrene that can be routinely used to study the differentiation of human pluripotent stem cell-derived neurons in vitro. Neurite outgrowth was significantly enhanced when neurons were grown in a three-dimensional environment compared to traditional flat surfaces and resulted in the formation of extensive neural networks. These data suggest that the topography within the culture environment can significantly alter cell development and will therefore be an important feature when investigating the potential of human stem cells.


Subject(s)
Cell Culture Techniques/instrumentation , Cell Culture Techniques/methods , Neurites/physiology , Neurons/cytology , Blotting, Western , Cell Differentiation , Cell Division , Cell Line, Tumor , Cells, Cultured , Humans , Microscopy , Microscopy, Electron, Scanning , Microscopy, Phase-Contrast , Neurons/metabolism , Polymers/chemistry , Polystyrenes/chemistry , Stem Cells/cytology , Time Factors
8.
Biochem Soc Trans ; 28(6): 680-1, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11171168

ABSTRACT

In an attempt to rationalize the relationship between structure and substrate selectivity of glycerol-3-phosphate acyltransferase (GPAT, 1AT, EC 2.3.1.15) we have cloned a number of cDNAs into the pET overexpression system using a PCR-based approach. Following assay of the recombinant enzyme we noted that the substrate selectivity of the squash (Cucurbita moschata) enzyme had altered dramatically. This form of GPAT has now been crystallized and its full three-dimensional structure elucidated. Since we now have two forms of the enzyme that display different substrate selectivities this should provide a powerful tool to determine the basis of the selectivity changes. Kinetic and structural analyses are currently being performed to rationalize the changes which have taken place.


Subject(s)
Cucurbitaceae/enzymology , Glycerol-3-Phosphate O-Acyltransferase/metabolism , Arabidopsis/enzymology , Cloning, Molecular , Crystallography, X-Ray , Escherichia coli , Glycerol-3-Phosphate O-Acyltransferase/chemistry , Glycerol-3-Phosphate O-Acyltransferase/genetics , Kinetics , Mutagenesis, Site-Directed , Polymerase Chain Reaction , Recombinant Proteins/chemistry , Recombinant Proteins/metabolism , Substrate Specificity
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