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E L Hogan

Publications and source records attributed to E L Hogan.

At least 55 records · Page 3Linked to original sources
PubMed1994

Branched monosialo gangliosides of the lacto-series isolated from bovine erythrocytes: characterization of a novel ganglioside, NeuGc-isooctaosylceramide.

We report the isolation, purification, and structural characterization of three monosialo gangliosides (G-1, G-2, and G-3) from bovine erythrocytes. Each of the purified compounds migrates as a single band on thin-layer chromatography in three solvent systems. All three gangliosides contain ceramide (Cer) as the lipid protein, with d18:1 sphingosine as the predominant long-chain base and with C18:0, C18:1, C20:0, C20:1, and C22:1 fatty acids, as determined by gas chromatography. The structural characterization of the carbohydrate moieties of G-1, G-2, and G-3 involved glycosyl composition analysis, methylation studies, sequential exoglycosidase hydrolysis, and one-dimensional 1H NMR spectroscopy of the native gangliosides. Furthermore, the oligosaccharides were released from the sphingolipids by endoglycoceramidase and fully sequenced by one- and two-dimensional 1H NMR spectroscopy in conjunction with fast-atom-bombardment mass spectrometry and exoglycosidase treatment. The structures are as follows: [formula: see text] Gangliosides such as G-1, G-2, and G-3, with branched oligosaccharide chains comprising a number of N-acetyllactosamine (Gal-GlcNAc) moieties, are abundant in erythrocytes from various mammalian species. The simultaneous occurrence of sialic acid and alpha-galactose as terminal sugars in these gangliosides, however, is relatively rare. Specifically, G-1 represents a ganglioside with a novel structure.

Amidohydrolases↗
PubMed1994

Peptide bond specificity of calpain: proteolysis of human myelin basic protein.

In order to determine the peptide bond specificity of calpain, human myelin basic protein (HMBP) was treated with purified calpain of bovine brain. Upon incubation, HMBP component I (HMBP-I) was degraded into several peptides as demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Component I was more susceptible to degradation than components II and III. HMBP degradation products were separated by high performance liquid chromatography (HPLC) and the cleavage sites in HMBP molecules were determined by peptide sequence analysis and by N- and C-terminal analyses. The major cleavage site was found to be 94Val-95Thr with several minor cleavages at 49Arg-50Gly, 18Ala-19Ser, 23His-24Ala, 27Gly-28Phe, 59Asp-60Ser, 70Gly-71Ser, 97Arg-98Thr, 110Ser-111Leu, 145Asp-146Ala, and 156Leu-157Gly. These results indicate that calpain is involved in the limited proteolysis of human myelin basic protein and prolonged incubation causes further digestion of the large peptides.

Amino Acid Sequence↗
PubMed1994

A revised structure for the disialosyl globo-series gangliosides of human erythrocytes and chicken skeletal muscle.

Disialosyl globo-series gangliosides have previously been isolated from chicken skeletal muscle (E. L. Hogan, R. D. Happel, and J.-L. Chien (1982) Adv. Exp. Med. Biol. 152, 273-278; S. Dasgupta, J.-L. Chien, E. L. Hogan, and H. van Halbeek (1991) J. Lipid Res. 32, 499-506) and human erythrocytes (S. K. Kundu, B. E. Samuelsson, I. Pascher, and D. Marcus (1983) J. Biol. Chem. 258, 13857-13866). In both cases, the structure of this ganglioside was proposed to be NeuAc alpha 2-->3(NeuAc alpha 2-->6)Gal beta 1-->3GalNAc beta 1-->3Gal alpha 1-->Gal alpha 1-->4Gal beta 1-->1Cer (V3NeuAcV6NeuAcGb5Cer). We have reinvestigated the human erythrocyte antigen and now propose an alternative structure differing in the location of the NeuAc alpha 2-->6 residue: NeuAc alpha 2-->3Gal beta 1-->3 (NeuAc alpha 2-->6)GalNAc beta 1-->3Gal alpha 1-->4Gal beta 1-->4Glc beta 1-->1 Cer (V3NeuAcIV6NeuAcGb5Cer). This novel structure is supported by results of 1H-NMR spectroscopy, negative ion fast atom bombardment mass spectrometry, and methylation linkage analysis with capillary gas chromatography--mass spectrometry in both electron impact and chemical ionization modes. Furthermore, based on new results from negative ion fast atom bombardment mass spectrometry and linkage analysis, we propose that the chicken skeletal muscle antigen also has this revised structure, differing only in ceramide composition. The terminal tetrasaccharide of these gangliosides is identical to that of GD1 alpha, NeuAc alpha 2-->3Gal beta 1-->3(NeuAc alpha 2-->6)GalNAc beta 1-->4Gal beta 1-->4Glc beta 1-->1 Cer(IV3NeuAcIII6NeuAcGg4Cer), previously identified in a rat ascites hepatoma cell line (T. Taki, Y. Hirabayashi, H. Ishikawa, S. Ando, K. Kon, Y. Tanaka, and M. Matsumoto (1986) J. Biol. Chem. 261, 3075-3078) and a murine lymphoma cell line with low metastatic potential (K. Murayama, S. B. Levery, V. Schirrmacher, and S. Hakomori (1986) Cancer Res. 46, 1395-1402), although they appear to be immunologically distinct.

Animals↗
PubMed1993

Effect of U-50488h, a selective opioid kappa receptor agonist, on vascular injury after spinal cord trauma.

U-50488H, a selective opioid kappa receptor agonist has been shown to be a neuroprotective agent in animal models of spinal cord injury. The mechanism of action of U-50488H is not known. Methylprednisolone, the only neuroprotective drug proven in patients with acute spinal cord injury may prevent the secondary injury after an initial trauma. Secondary vascular injury develops after experimental spinal cord trauma. In this study we examined the effects of U-50488H on post-traumatic vascular injury based on the measurement of vascular permeability, edema and neutrophil infiltration in a rat spinal cord injury model. Vascular permeability was assessed by vascular extravasation of fluorescein isothiocyanate conjugated dextran (FITC-D), a macromolecular tracer. Tissue edema was determined by percentage water content and neutrophil infiltration by myeloperoxidase (MPO) activity, a marker enzyme for neutrophils. U-50488H at doses of 5, 10, 20 and 40 mg/kg i.p. administered twice (0.5 h before and 0.5 h after trauma) reduced vascular permeability in a dose-dependent manner. More frequent dosing (10 mg/kg, 0.5 h before and 0.5, 2, 8 and 22 h after injury) reduced vascular permeability 24 h after injury. U-50488H also reduced edema formation but did not affect neutrophil infiltration. Results from this study raise the possibility that the neuroprotective effect of U-50488H involves a secondary vascular event.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1993

Calcium-activated neutral proteinase (calpain) in rat brain during development: compartmentation and role in myelination.

The activity of both forms (microM and mM Ca(2+)-sensitive) of calcium-activated neutral proteinase (calpain) was determined in developing rat brain. Triton X-100 did not affect mcalpain activity at the earlier ages (1-5 days postpartum) whereas mcalpain activity significantly increased at 16 days and older. The mcalpain activity in brain was negligible at earlier ages (1-7 days) and the peak activity occurred between 16 and 30 days after birth. The peak activity of mcalpain in myelin was found between 16 and 30 days of age and myelin from rats older than 30 days contained 40-50% of the brain mcalpain activity. In contrast, 70-80% of the brain mcalpain activity was in cytosol at younger ages (1-10 days) and decreased to 30% with increasing age (90 days). On the other hand, mu calpain was found mainly (65-75%) associated with a membrane fraction (microsomes) before 10 days and the majority of the activity was found in cytosol (68%) between 16 and 30 days. Immunoblot studies revealed mcalpain in both myelin and cytosol from developing rat brain. These results indicate that mcalpain is present in myelin and suggest that it may be involved in the formation of myelin sheath.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗
PubMed1993

Effects of fish oil supplementation on acute ischemic brain injury in the rat.

We studied the effects of long-term fish oil (FO) dietary supplementation on brain edema, polymorphonuclear neutrophil (PMN) infiltration, and infarct size in a rat stroke model. Rats were given regular rat chow with or without FO supplement (20% of total calories) for 7 weeks. Body weight did not differ between the two groups. In the FO group, an increase in eicosapentaenoic acid and a decrease in arachidonic acid content in hepatic phospholipids were significant in the phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine but not in the phosphoinositol fraction. Platelet activity reflected by serum thromboxane B2 levels was reduced in the FO group. Postischemic brain edema and PMN infiltration were not different between the two groups. The infarct volume was significantly greater in the FO group (controls: 96 +/- 7 mm3, n = 49; FO group: 124 +/- 6 mm3, n = 53; p = 0.0036). The greater ischemic brain injury in the FO-supplemented animals is probably related to the intraischemic hyperglycemia, which was worse in the FO group than in the control group (controls: 265 +/- 19 mg/dl, n = 14; FO group: 340 +/- 18 mg/dl, n = 16; p = 0.0079).

Animals↗
PubMed1993

Molecular characterization of gangliotetraosylceramide (GA1) in normal human brain and its developmental change.

Several neutral glycosphingolipids have recently been purified from normal human brain to the criterion of migration as homogeneous bands in two different solvent systems. One of these has been permethylated and analyzed by gas chromatography. Stepwise specific exoglycosidase hydrolysis confirms its structure as GAl or Gal beta 1-->3GalNAc beta 1-->4Gal beta 1-->4Glc beta 1-->1Cer. The neutral glycosphingolipids of rat CNS have been examined in order to investigate changes in GA1 during critical ages of brain development. Employing the highly sensitive techniques of digoxigenin (DIG) immunostaining and Fluorescence Assisted Carbohydrate Electrophoresis (FACE), we have identified several previously uncharacterized long chain neutral glycosphingolipids in brain and myelin. A major band with Rf close to that of nLcOse5Cer purified from bovine erythrocytes has been identified as GA1 by immuno-TLC using mono-specific polyclonal anti-GA1 antisera. It appears in 5 day-postpartum (P5) developing brain, increases until 21 days (P21) and subsequently declines. This phasic change along with changes in other nonsialylated glycosphingolipids of the developing brain strongly suggests that GA1 and other neutral glycosphingolipids may play a mediator role(s) in brain development and/or myelinogenesis.

Adult↗
PubMed1992

Ganglio-N-tetraosylceramide (GA1) of bovine and human brain. Molecular characterization and presence in myelin.

During our studies of bovine brain neutral glycosphingolipids (Ngsl's), we have purified a compound that co-migrates on thin-layer chromatogram with standard GA1 (purified by acid hydrolysis of GM1) and close to penta- (nLc5Cer) glycosylceramide from bovien erythrocytes. The structure of the purified Ngsl from brain has been established by permethylation and by stepwise exoglycosidase hydrolysis. 600 MHz 1H NMR spectroscopy of the oligosaccharide obtained from the Ngsl after endoglycoceramidase hydrolysis confirms the structure as ganglio-N-tetraosylceramide (GgOse4Cer or GA1) as Gal beta 1----3GalNAc beta 1----4Gal beta 1----4Glc beta 1----1Cer. We have identified GA1 in bovine, rat and human brain and myelin by TLC-immunostaining with monospecific anti-GA1 antiserum.

Animals↗
PubMed1992

Calcium-activated neutral proteinase (calpain) activity in C6 cell line: compartmentation of mu and m calpain.

Calcium-activated neutral proteinase (calpain) activity was determined, including in cytosol and membrane fractions, in rat glioma C6 cell line. The mu and m forms of calpain were separated by DEAE and phenylsepharose column chromatography and with removal of the endogenous inhibitor calpastatin. C6 cells contained more mcalpain than the mu isoform. More than 70% of mcalpain activity was membrane-associated and 20% was cytosolic. Isolated plasma membrane also contained 69% of the mcalpain activity. In contrast, approximately 80% of mucalpain activity was cytosolic and 16% was membranous. Half-maximal activity for mu and mcalpain was obtained at 1 microM and 0.2 mM CaCl2, respectively. Trypsin dissociation of cells reduced activity. Triton X-100 stimulated mcalpain activity of the whole homogenate and the membrane pellet but not of the cytosol. Activity of the myelin marker enzyme adenosine 2'3'-cyclic nucleotide 3'-phosphohydrolase (CNPase), was also found in C6 cells. The identification of calpain and CNPase in C6 cells is in keeping with an interpretation that C6 differentiation resembles, at least in part, that of the myelin-forming oligodendroglial cells.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗
PubMed1992

Susceptibility of myelin proteins to a neutral endoproteinase: the degradation of myelin basic protein (MBP) and P2 protein by purified bovine brain multicatalytic proteinase complex (MPC).

Multicatalytic proteinase complex (MPC) was isolated from bovine brain and the susceptibility of myelin basic protein (MBP) and P2 protein of bovine central and peripheral nervous system was examined. SDS-polyacrylamide electrophoretic analysis of purified MPC revealed protein bands of molecular weight ranging from 22-35 kDa. The enzyme is activated by SDS at a concentration less than 0.01%. Upon incubation with MPC, purified MBP and P2 proteins were degraded into smaller fragments. There was a 57% and 100% loss of MBP at 2 and 6 hours of incubation. The P2 protein which is not susceptible to any endogenous non-lysosomal enzyme thus far studied was digested into small peptide fragments only in the presence of SDS (0.01%) and not in its absence. These results indicate that MPC which is active at physiological conditions may have a role in the turnover of myelin proteins and in demyelinating diseases.

Amino Acid Sequence↗
PubMed1992

Effects of detergents on Ca(2+)-activated neural proteinase activity (calpain) in neural and non-neural tissue: a comparative study.

Calcium activated neutral proteinase (mcalpain) activity was determined in brain and other tissue of rat. More than 60% of the brain mcalpain activity was present in the particulate fraction while only 30% was in cytosol. In contrast, particulate fractions of liver, kidney, muscle, and heart contained about 8-12% of tissue mcalpain activity while 88% was present in cytosol. Removal of the endogenous inhibitor calpastatin increased the tissue mcalpain activity severalfold. Triton X-100 and deoxycholate (DOC) stimulated the neural calpain activity by ten-fold while activity in non-neural tissue was unaffected. Incubation with other detergents, e.g. Triton N-57 and thioglucopyranoside, stimulated brain calpain activity five-fold while Brij-35 did not have any effect. Sodiumdodecylsulphate (SDS), on the other hand, inhibited the enzyme activity. Brain contained the lowest calpain activity compared to non-neural tissue. The calpain activity in muscle, kidney and heart was three-fold greater than liver. Immunoblot identification of the enzyme revealed that calpain was predominantly in the particulate fraction and less in cytosol of brain while it was present mainly in cytosol and less in the pellet fractions of non-neural tissue.

Animals↗
PubMed1992

Protective effect of methylprednisolone on vascular injury in rat spinal cord injury.

High-dose methylprednisolone (MP) given to patients within 8 h of traumatic spinal cord improved neural function at 6 and 12 months, suggesting a probable secondary injury process that may be amenable to therapeutic intervention. Vascular injury plays an important role in the secondary injury process of CNS trauma. We have examined the effect of MP on vascular changes, including tissue edema, vascular permeability, and polymorphonuclear (PMN) cell infiltration in a rat model of spinal cord impact injury. MP significantly reduced extravasation of fluorescein isothiocyanate dextran (FITC-D), a macromolecular tracer, by 64.3% and 50.7% with trauma forces of 20 and 40 g-cm, respectively, when MP was administered IV immediately after trauma at a bolus of 165 mg/kg, with a subsequent continuous MP infusion at 31.5 mg/kg/h for 23 h. MP reduced the water content in the 40 g-cm traumatic cord lesion to 73.0% compared to the traumatic control (74.3%, p < 0.001) at the same schedule of large dose 24-h infusion. The same doses of MP showed a trend to decrease the extent of neutrophil infiltration as determined by myeloperoxidase (MPO) activity, but the change was not significant. MP had little effect in decreasing FITC-D extravasation and cord edema when given at a lower dose (bolus of 30 mg/kg with continued infusion of 1.3 mg/kg/h for 23 h). MP did not reduce extravasation of FITC-D and edema when administered IV as one bolus injection at high (165 mg/kg) or low (30 mg/kg) doses.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Receptor-linked hydrolysis of phosphoinositides and production of prostacyclin in cerebral endothelial cells.

The receptor agonist-mediated hydrolysis of phosphoinositides and production of prostacyclin were studied in murine cerebral endothelial cells (MCEC). Of 11 neurotransmitters and neuromodulators examined, carbachol, noradrenaline (NE), bradykinin, and thrombin significantly increased 3H-inositol phosphate accumulation in the presence of LiCl (20 mM). The maximal stimulation of [3H]inositol monophosphate ([3H]IP1) reached approximately 11, 11, seven, and four times the basal levels for carbachol, NE, bradykinin, and thrombin, respectively. The EC50 values of IP1 accumulation for carbachol and NE were 34 and 0.16 microM, respectively. The muscarinic antagonists, atropine and pirenzepine, blocked the carbachol-induced IP1 accumulation with Ki values of 0.3 and 30 nM, respectively. The adrenergic antagonist, prazosin, blocked NE-induced IP1 accumulation with a Ki of 0.1 nM. The calcium ionophore A23187, histamine, glutamate, vasopressin, serotonin, platelet activating factor, and substance P did not stimulate IP1 accumulation. A23187, bradykinin, and thrombin stimulated prostacyclin release to approximately four, four, and two times the basal levels, respectively, whereas carbachol and NE had little effect upon prostacyclin release. These results suggest that the activation of phospholipase C and of phospholipase A2 in MCEC are regulated separately.

Animals↗
PubMed1991

Calcium-activated neutral proteinase (CANP; calpain) activity in Schwann cells: immunofluorescence localization and compartmentation of mu- and mCANP.

Calcium-activated neutral proteinase (CANP) activity was determined in cytosolic and membranous subcellular fractions of transformed Schwann cells (tSc). The muM and mM Ca(2+)-sensitive (mu- and mCANP) forms of CANP were separated by DEAE and phenyl Sepharose column chromatography, the latter step enabling removal of the endogenous inhibitor calpastatin. The tSc contained more muCANP than the mM isoform. More than 75% of mCANP activity was membrane-associated and 20% was cytosolic. In contrast, approximately 80% of muCANP was cytosolic and 15% was membranous. Triton X-100 stimulated activity of the whole homogenate and of the membrane pellet but did not stimulate CANP activity in the cytosolic fraction. Immunohistochemical distribution of mM enzyme was studied in both fixed and permeabilized tSc with cytosolic (anti-cyt-mCANP) and myelin (anti-my-mCANP) antibodies. Live cells (non-permeabilized) stained with anti-my-mCANP had a single filamentous ring circumscribing individual cells. Permeabilized cells treated with anti-my-mCANP had immunoreactive deposits throughout the intracellular space but sparing the perinuclear region. No immunohistochemical staining was detected when live cells were exposed to anti-cyt-mCANP whereas permeabilized cells had extensive intracellular staining with the most intense immunoreactivity in the perinuclear region. Our results indicate that both forms of CANP are present in tSc and that the activity of most of the muCANP is cytosolic while mCANP is particulate.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗
PubMed1991

Differential regulation of myelin gene expression in SV40 T antigen-transfected rat glioma C6 cells.

Rat glioma C6 cells were stably transfected with a pSV3-neo plasmid containing SV40 T antigen gene, and geniticin-resistant transfectants (designated C6T cells) were cloned. The C6T cells grew as well-defined foci of cells showing squamous or irregular morphology. The doubling time for transfected cells was reduced by approximately 40% as compared to control C6 cells. The transfection with T-antigen also affected the expression of genes coding for structural myelin proteins and for myelin-associated enzymes. The steady-state level of proteolipid protein (PLP)-specific mRNA in C6T cells was 44% lower than in parental C6 cells. On the other hand, the transfection upregulated the expression of myelin-associated glycoprotein (MAG) by 153%. The activity of 2':3' cyclic AMP phosphodiesterase (CNP) was increased by approximately 80% in the C6T cells as compared to untransfected, control cells. The activity of calcium-activated neutral proteinase (CANP) was also significantly elevated in the transfectants by approximately 50% and 220% for millimolar and micromolar form respectively. The results indicate that T antigen affects the expression of myelin genes, although, individual genes appear to be differently regulated implying the existence of several independent regulatory mechanisms.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1991

Xanthine oxidase in experimental spinal cord injury.

The excessive generation of free radicals is thought to be one of the major mechanisms leading to tissue injury in various pathological conditions, including ischemia, inflammation, and trauma. Conversion of xanthine dehydrogenase (XDH) to xanthine oxidase (XO) contributes to the formation of superoxide, an oxygen radical. We measured XDH and XO activity using a newly developed fluorometric assay in an experimental spinal cord injury model in rats. XO activity increased by more than 100% 4 h after spinal cord trauma. Total (XDH + XO) activity also increased by 96% during the same period. Allopurinol, an inhibitor of XO (100 mg/kg/day x 2 days, i.p.), completely inhibited plasma and spinal cord XO activity but did not affect posttraumatic edema determined by water content or polymorphonuclear (PMN) cell infiltration reflected by myeloperoxidase (MPO) activity in traumatized spinal cord. These results indicate that XDH conversion to XO may not be the major mechanism of oxygen radical formation in the pathogenesis of vasogenic edema or inflammatory response in this experimental spinal cord injury model in rats.

Allopurinol↗