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Biomedical subjects

S Roberts

Publications and source records attributed to S Roberts.

At least 163 records · Page 9Linked to original sources
PubMed1996

Stimulation of actin stress fibre formation mediated by activation of phospholipase D.

BACKGROUND: Agonist-stimulated phospholipase D (PLD) catalyzes the hydrolysis of phosphatidylcholine, generating the putative messenger phosphatidate (PA). Proposed functions for PA, and hence for PLD, include kinase activation, the regulation of small molecular weight GTP-binding proteins, actin polymerization and secretion. It has not been possible to define a physiological function for PLD activation as it is generally stimulated together with other signalling pathways, such as those involving phospholipases A2 and C, phosphatidylinositide (PI) 3-kinase and the p21(ras)/mitogen-activated protein (MAP) kinase cascade. RESULTS: We report that, in porcine aortic endothelial (PAE) cells, lysophosphatidic acid (LPA) stimulated PLD activity and rapidly generated PA in the absence of other phospholipase, PI 3-kinase or MAP kinase activities. PLD activation was controlled by a tyrosine kinase-regulated pathway. LPA also stimulated actin stress fibre formation, but was inhibited by butan-1-ol; the alcohol also reduced the accumulation of PA. The addition of PA to cells did not stimulate PLD activity, but did cause stress fibre formation in a manner that was insensitive to butan-1-ol. Stimulation of stress fibre formation by LPA and PA was sensitive to genistein, and was inhibited by micro-injection of the Rho-inhibiting C3 exotoxin into PAE cells. CONCLUSIONS: This study provides the first clear demonstration of a physiological role for PLD activity. In PAE cells, the stimulation of actin stress fibre formation was a consequence of PA generation and, therefore, PLD activation. The results suggest that PA generation is upstream of Rho activation, and imply a role for PLD in the regulation of Rho-mediated pathways.

Actins↗
PubMed1996

Transport properties of the human cartilage endplate in relation to its composition and calcification.

STUDY DESIGN: The transport properties of solutes of different sizes and conformations were studied in cartilage endplates. OBJECTIVES: The results were correlated with the composition of the cartilage matrix to determine if a relationship existed between this and the movement of molecules within it. SUMMARY OF BACKGROUND DATA: Solute transport through the hyaline cartilage endplate is important not only for the physiologic and metabolic processes of that tissue, but also for those of the adjacent intervertebral disc. Movement of solutes depends on solute size, shape or charges, and the composition of the matrix itself. Changes in composition of the cartilage endplate, such as those that occur in degeneration or scoliosis, may affect transport. METHODS: Partition and diffusion coefficients of solutes ranging in molecular weight from 115 to 70,000 d have been measured on cores of cartilage endplate. Transport properties were assessed in relation to core composition. RESULTS: The shape and size of the solutes were found to affect their transport through cartilage matrix, with larger molecules being more highly excluded and diffusing more slowly. Long-chain polymers were able to penetrate the matrix less readily than the more globular molecules. The more hydrated the matrix, the higher the degree of penetration and the more easily solutes could move, in contrast to the inverse relationship between the other components of the matrix and solute transport. With increased proteoglycan, collagen, or calcification in the tissue, there was greater restriction of solute movement. CONCLUSIONS: The proteoglycans normally found in the endplate regulate movement of solutes into and out of the disc. It has been shown previously that removal of proteoglycans from the endplate accelerates the loss of proteoglycans from the nucleus. Hence, a major function of the cartilage endplate may be to prevent fragments of osmotically active proteoglycans from leaving the disc.

Adolescent↗
PubMed1996

The affinity of allergen specific IgE and the competition between IgE and IgG for the allergen in Amb a V sensitive individuals.

We have calibrated a solid state RAST assay with affinity purified allergen-specific IgE. We then utilized the calibrated assay to measure the average affinity of individual IgE-containing sera in terms of the average association constant < K > for purified allergen Amb a V. The binding data yielded linear reciprocal plots indicating that the range of affinities of the responding clones was narrow. The range of the average association constant for the IgE-Amb a V complex was 0.9-26 x 10(10) M-1. The average affinity of the corresponding IgG response in the same individual, estimated by inhibition studies of IgE binding, was 10(7) M-1 in one case and lower than 10(6) M-1 in all the other cases.

Allergens↗
PubMed1996

A review of parametric modelling techniques for EEG analysis.

This review provides an introduction to the use of parametric modelling techniques for time series analysis, and in particular the application of autoregressive modelling to the analysis of physiological signals such as the human electroencephalogram. The concept of signal stationarity is considered and, in the light of this, both adaptive models, and non-adaptive models employing fixed or adaptive segmentation, are discussed. For non-adaptive autoregressive models, the Yule-Walker equations are derived and the popular Levinson-Durbin and Burg algorithms are introduced. The interpretation of an autoregressive model as a recursive digital filter and its use in spectral estimation are considered, and the important issues of model stability and model complexity are discussed.

Biomedical Engineering↗
PubMed1996

Morphological, molecular, and functional heterogeneity of cholangiocytes from normal rat liver.

BACKGROUND & AIMS: While the lobular heterogeneity of hepatocytes is established, limited data suggest that bile duct epithelial cells, of cholangiocytes, are heterogeneous along the normal intrahepatic biliary tree. Thus, we tested the hypothesis that cholangiocytes are structurally and functionally heterogeneous in the biliary tract of normal rats. METHODS: A series of in situ and in vitro experiments was performed in normal rats using complementary morphometric, molecular, and functional approaches. RESULTS: In situ morphometry showed that (1) intrahepatic bile ducts are heterogeneous in external diameter (5-200 mum); (2) individual cholangiocytes lining bile ducts are heterogeneous in area (3-80 mum2); and (3) a significant relationship exists between bile duct diameter and cholangiocyte area. Using a novel approach developed by us, we isolated three subpopulations of small, medium, and large cholangiocytes and compared them at the molecular and functional level. The expression of two cholangiocyte-specific genes (gamma-glutamyl transpeptidase and cytokeratin 19) was similar among the three subpopulations. In contrast, secretion receptor, Cl-/HCO3- exchanger, and cystic fibrosis transmembrane conductance regulator messenger RNAs were differentially expressed, being present on medium and large but not small cholangiocytes. At the functional level, adenosine 3', 5'-cyclic monophosphate and intracellular pH responses and Cl-/HCO3- exchanger activity was enhanced by secretion in medium and large but not small cholangiocytes. CONCLUSIONS: These data indicate that cholangiocytes are morphologically and functionally heterogeneous along the normal intrahepatic biliary tree and suggest that secretion-regulated transport of water and electrolytes occurs principally in medium and large ducts.

Animals↗
PubMed1996

A new approach to the analysis of the human sleep/wakefulness continuum.

The conventional approach to the analysis of human sleep uses a set of pre-defined rules to allocate each 20 or 30-s epoch to one of six main sleep stages. The application of these rules is performed either manually, by visual inspection of the electroencephalogram and related signals, or, more recently, by a software implementation of these rules on a computer. This article evaluates the limitations of rule-based sleep staging and then presents a new method of sleep analysis that makes no such use of pre-defined rules and stages, tracking instead the dynamic development of sleep on a continuous scale. The extraction of meaningful features from the electroencephalogram is first considered, and for this purpose a technique called autoregressive modelling was preferred to the more commonly-used methods of band-pass filtering or the fast Fourier transform. This is followed by a qualitative investigation into the dynamics of the electroencephalogram during sleep using a technique for data visualization known as a self-organizing feature map. The insights gained using this map led to the subsequent development of a new, quantitative method of sleep analysis that utilizes the pattern recognition capabilities of an artificial neural network. The outputs from this network provide a second-by-second quantification of the sleep/wakefulness continuum with a resolution that far exceeds that of rule-based sleep staging. This is demonstrated by the neural network's ability to pinpoint micro-arousals and highlight periods of severely disturbed sleep caused by certain sleep disorders. Both these phenomena are of considerable clinical value, but neither are scored satisfactorily using rule-based sleep staging.

Adult↗
PubMed1996

Problem of diagnosis in postmortem brain studies of schizophrenia.

OBJECTIVE: The purpose of this study was to determine 1) the reliability of diagnoses of schizophrenia at coronal autopsy and 2) the degree to which the use of different diagnostic instruments for schizophrenia would affect postmortem brain research. METHOD: Eighty-three subjects, recorded at coronal autopsy as having had schizophrenia, were referred for neurochemistry studies. The diagnoses reported to the state coroner's office were reevaluated by a review of psychiatric case histories by clinicians using semistructured assessment and diagnostic criteria. RESULTS: The application of DSM-III-R, Research Diagnostic Criteria (RDC), ICD-10, Schneiderian, and Feighner criteria to the diagnosis of the 83 subjects revealed that 63.9%, 48.2%, 48,2%, 43.4%, and 42.2%, respectively, met the criteria for schizophrenia. Highest concordance was between the RDC and ICD-10 systems, while lowest concordance was between the RDC and Schneider systems. CONCLUSIONS: These data suggest that unless carefully reviewed, diagnosis may be a major confounding factor in postmortem studies of brain tissue from subjects with schizophrenia.

Adult↗
PubMed1995

Mechanoreceptors in intervertebral discs. Morphology, distribution, and neuropeptides.

STUDY DESIGN: The present study investigated the occurrence and morphology of mechanoreceptors in human and bovine intervertebral discs and longitudinal ligaments. OBJECTIVE: To determine the type and frequency of mechanoreceptors present in intervertebral discs and anterior longitudinal ligaments in two patient groups, those with low back pain and those with scoliosis. Bovine coccygeal discs were examined. SUMMARY OF BACKGROUND DATA: Nerves have been described in intervertebral tissues, but there is little information on the endings of these nerves and their receptors, stimulation of which can cause a nerve impulse. METHODS: The presence of mechanoreceptors were investigated by immunolocalization of nerves and neuropeptides. By examining sequential sections, the frequency of receptors was assessed. RESULTS: Immunoreactivity to neural antigens showed mechanoreceptors in the anulus fibrosus and longitudinal ligaments of bovine and human specimens. Their morphology resembled Pacinian corpuscles, Ruffini endings, and, most frequently, Golgi tendon organs. They were found in 50% of discs investigated from patients with low back pain and in 15% of those with scoliosis. CONCLUSIONS: Mechanoreceptors were found in the outer 2-3 lamellae of the human intervertebral disc and anterior longitudinal ligament. Physiologic studies in other tissues indicate that these provide the individual with sensation of posture and movement, and in the case of Golgi tendon organs, of nociception. In addition to providing proprioception, mechanoreceptors are thought to have roles in maintaining muscle tone and reflexes. Their presence in the intervertebral disc and longitudinal ligament can have physiologic and clinical implications.

Adolescent↗
PubMed1995

Rapid turnover and impaired cell-surface expression of the human folate receptor in mouse L(tk-) fibroblasts, a cell line defective in glycosylphosphatidylinositol tail synthesis.

The human folate receptor (hFR) is a plasma membrane protein that is anchored to the membrane via a glycosylphosphatidylinositol (GPI) tail in some cell types. The KB hFR cDNA sequence predicts a hydrophobic, alpha-helical 31-residue carboxyl terminus that is thought to be the signal for cleavage and attachment of the GPI tail. Alternatively, this region may serve as a transmembrane domain if GPI attachment is not efficient. In this study, we investigated the latter possibility by expressing the hFR in L(tk-) cells, cells that are unable to synthesize GPI tails for attachment to membrane proteins. We also transfected the same hFR cDNA into Chinese hamster ovary (CHO) cells, cells that can anchor proteins by either a GPI tail or a transmembrane domain. Neither parental cell line expresses detectable levels of folate receptor as determined by folic acid binding assays, Western analysis, or Northern analysis. In L(tk-) cells, we found that the recombinant hFR is not expressed on the cell surface, but is rapidly degraded (t1/2 < or = 4 h). Most (> 95%) of the recombinant hFR remains Endo H sensitive, suggesting retention in the endoplasmic reticulum. In contrast, transfected CHO cells express functional hFR protein at the cell surface, the half-life of the hFR is long (t1/2 > or = 24 h), and the Endo H glycosylation pattern of the recombinant hFR is consistent with normal processing through the Golgi apparatus. Therefore, in the absence of a GPI tail, the hFR is not sorted to the cell surface and the incompletely processed hFR protein is unstable.

Animals↗
PubMed1995

The high levels of p53 present in adenovirus early region 1-transformed human cells do not cause up-regulation of MDM2 expression.

The cellular protein MDM2 can bind to the tumor suppressor gene product p53 and abrogate its transcriptional activity. In addition, p53 can regulate expression of the mdm2 gene. We and others have previously shown that p53 is present at high levels in adenovirus-transformed cells which express the larger E1B protein. In view of these observations the expression of MDM2 in a panel of adenovirus transformed human cell lines has been examined. Two major species (98K and 80K) were detected, together with a number of minor species of higher and lower molecular weight. While there was little variation in levels of 98K protein between cell lines, appreciable differences in the expression of the 80K component were apparent. There was no correlation between MDM2 and p53 expression in any of the adenovirus transformants, nor with the viral proteins expressed. The pattern and level of MDM2 detected was similar to that seen in human tumor cell lines and in human fetal tissue. Northern blot analysis suggested that MDM2 expression was regulated at the transcriptional level. Stable interactions were observed between p53 and MDM2 in the adenovirus-transformed cell lines and in Ad5 E1 HEK 293 cells a ternary complex of p53, MDM2, and the Ad5 E1B 58K protein was demonstrated. In view of the lack of correlation between the level of p53 and MDM2 in adenovirus E1-transformed cells, the capacity of p53 to cause transcriptional activation was assessed using transfected CAT constructs linked to p53 responsive elements. p53 transcriptional activity was similar in all of the cell lines examined and did not correlate with protein expression. It is concluded, on the basis of all of these data, that the high concentrations of p53 found in adenovirus transformants are not transcriptionally active and have no influence on MDM2 expression. However, when expression of p53 was increased following infection with mutant adenoviruses, which do not express the larger E1B proteins, there was an appreciable increase in p53 transcriptional activity and in the levels of all of the MDM2 components.

Adenovirus E1B Proteins↗