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Biomedical subjects

E L Hogan

Publications and source records attributed to E L Hogan.

At least 109 records · Page 6Linked to original sources
PubMed1986

Sialylation of lacto-N-neohexaosylceramide by sialyltransferase from embryonic chicken muscle.

A sialyltransferase which catalyzes the in vitro biosynthesis of N-acetylneuraminosyllacto-N-neohexaosylceramide from lacto-N-neohexaosylceramide and CMP-NeuAc has been examined in embryonic chicken breast muscle. The maximum enzyme activity was observed in 11-12-day-old embryos. The enzyme has optimum activity at pH 6.8 in the presence of Triton CF-54 and Mg2+. The apparent Km values for lacto-N-neohexaosylceramide and CMP-NeuAc were 0.9 and 0.67 mM, respectively. The enzymic product was characterized by TLC, neuraminidase hydrolysis and permethylation analysis. The structure was identical to authentic N-acetylneuraminosyllacto-N-neohexaosylceramide from chicken muscle. In addition, a disialo derivative has been detected that constitutes 15% of the total radioactivity incorporated. The two sialic acids connected by sialosyl-sialosyl linkage were attached to the terminal galactose residue. To our knowledge, this is the first report of biosynthesis of this disialo compound.

Animals↗
PubMed1986

Phospholipid biosynthesis during normal and dystrophic avian muscle cell differentiation in culture.

The biosynthesis of phospholipids during differentiation of normal and dystrophic muscle cells was studied using [32Pi] added to primary culture media. Labeled phosphatides were separated by two-dimensional thin-layer chromatography, exposing the plates to HCl vapors between solvents to cleave the alk-1-enyl bonds. Phosphatidylethanolamine labeling did not vary during myotube formation. Although lower labeling of phosphatidylcholine was observed in dystrophic myoblasts at the single cell stage, no differences were observed between normal and dystrophic cultures after cell contact and myotube formation. A sharp decrease in phosphoinositide content was observed as myoblasts first achieved cell contact and continued to decline slowly as myotubes developed. During myogenesis, phosphatidylcholine content gradually increased, from 18.4% of the total labeled phospholipid in dividing myoblasts to 48.6% of the labeled phospholipids in fully differentiated myotubes.

Animals↗
PubMed1986

Proteolysis in quaking mouse brain and spinal cord.

Six proteolytic enzymes were assayed for activity in quaking CNS in examining the hypothesis that increased proteolytic activity contributes to the hypomyelination characteristics of this mutant. Cathepsin B-like enzyme, cathepsin D, neutral proteinase, calcium-activated neutral proteinase, prolyl endopeptidase, and diaminopeptidase II were assayed in whole homogenate of brain or spinal cord and each was found to have activity similar to that in normal mice. These results do not support a relationship between proteolysis and the genetic defect and suggest that other factors should be investigated to delineate the pathogenesis of this mutant.

Animals↗
PubMed1986

Synthesis and acylation of myelin proteolipid protein in quaking mouse brain.

The synthesis and acylation of proteolipid proteins where investigated in tissue slices prepared from 19-day-old quaking and normal littermate mouse brain. The mutant CNS had a normal rate of total protein synthesis but synthesis of the myelin-specific proteins, proteolipid protein (PLP) and intermediate protein (IP), was impaired to approximately 50% of control. The acylation of myelin proteins with labeled palmitate was reduced to only about 20% of control. The acylation of two nonmyelin proteolipid proteins was also significantly reduced. The incorporation of palmitate into phospholipids was identical in control and mutant. The results indicate impaired synthesis of PLP and IP and a general deficit in protein acylation in quaking brain.

Acylation↗
PubMed1986

Lipid content of swine influenza and other vaccines.

An analysis of the lipids in swine influenza vaccines was performed, comparing six different lots of swine influenza, other influenza and noninfluenza vaccines. Cholesterol content and phospholipid content varied greatly, but there were no major differences between the types of vaccines. Appreciable amounts of phosphatidylethanolamine were found in only one swine influenza vaccine. The major phospholipids of influenza vaccines were phosphatidylcholine, sphingomyelin and phosphatidic acid. A detectable amount of phosphatidylserine was not found in any swine influenza vaccine, but was present in two of three nonswine influenza vaccines. Only two of six swine influenza vaccines showed trace amounts (less than 0.5 microgram/ml) of ganglioside (GM3). However, larger quantities of galactocerebroside were found (2.24-6.43 micrograms/ml) in all influenza vaccines examined, including swine influenza vaccines.

Cholesterol↗
PubMed1986

Increased thromboxane level in experimental spinal cord injury.

An increased accumulation of tissue thromboxane A2 occurred shortly after spinal cord injury. Prostacyclin formation was not affected. The magnitude of the increase in thromboxane and the extent of post-traumatic vascular damage as determined by extravasation of 125I-labeled human serum albumin were both dependent on the degree of injury. These findings raise the possibility that activation of arachidonic acid metabolism with a preponderance in thromboxane formation may contribute to microvascular injury in experimental spinal cord contusion.

6-Ketoprostaglandin F1 alpha↗
PubMed1986

Proteolytic enzymes in experimental spinal cord injury.

Experimental spinal cord injury was produced in rats by dropping a 10 g weight from 30 cm upon dura-invested exposed spinal cord. Proteolytic activities at neutral (pH 7.6) and acid (pH 5.5 and 3.6) pH were determined in whole homogenate and the cytosolic fraction of the lesion (lumbar) and cervical control segments. The enzyme activity was monitored by SDS-PAGE analysis of the extent of substrate myelin basic protein (MBP) degradation. Activities (neutral and cathepsin B-like) in the sham-operated spinal cord were lower than those of cervical autologous control at 24 h after injury. The increase in neutral proteinase activity was progressive and greater in the lesion than the autologous control. A 61.5% +/- 3.5 loss of MBP was observed at 2 h following injury and increased at 24 h (78.2% +/- 3.4). The loss of MBP coincided with the appearance of several low molecular weight peptides. The cathepsin B-like and cathepsin D activities were also increased in the lesion but to a lesser extent than the neutral proteinase. The neutral proteinase and cathepsin B-like activity were inhibited by leupeptin and not by pepstatin while the converse obtained for cathepsin D activity. The release of neutral proteolytic activity which is nonlysosomal in origin suggests a novel hypothesis for the mechanism of traumatic axon-myelin injury.

Animals↗
PubMed1986

Thromboxane, prostacyclin, and leukotrienes in cerebral ischemia.

Our understanding of the biochemistry and biologic actions of AA metabolites has been greatly expanded in recent years. The discoveries of TXA2, PGI2, and LTs have fostered new concepts of the pathophysiology of cerebral ischemia. New approaches to treatment of ischemia include seeking an optimal dose of aspirin, developing drugs that selectively inhibit or antagonize TXA2 or LTs, and administering PGI2 or its analogues. Altering the dietary content of essential fatty acids for prophylaxis is also being studied. Though the results of this thrust are still preliminary, the exploration of these therapeutic strategies in cerebrovascular disorders based on further understanding of the pathophysiologic roles of TXA2, PGI2, LTs and probably other AA metabolites is anticipated with some optimism.

Blood Platelets↗
PubMed1985

Quantitation of the in vitro neuroblastoma response to exogenous, purified gangliosides.

Individual ganglioside species (possessing the gangliotetrose oligosaccharide) were purified from bovine brain gray matter and applied in varying concentrations to the culture medium of mouse neuroblastoma cells (N2A) in vitro. After 48 hr of incubation, the cells were stained, and the neuritogenic response quantitated with a video analysis system, employing a program to measure three parameters of neuroblastoma differentiation: neurites per cell (sprouting), neurite length (extension), and degree of neurite branching (arborization). All the individual gangliosides tested promoted neurite extension in a dose-dependent fashion. Asialogangliosides ("neutral" glycosphingolipids) were without effect, which suggests that sialic acid (N-acetylneuraminic acid) is necessary to elicit this cellular response. With increasing concentrations of GM1 (5 to 500 micrograms/ml), the average cellular neurite length increased significantly, whereas the number of neurites per cell decreased. With the trisialoganglioside GT1b, neurite length did not increase to the extent seen with GM1, but an increase in the number of neurites per cell (sprouting) and branch points per neurite (arborization) was observed. These results suggest that the in vitro neuronal response to exogenous gangliosides may combine specific responses to individual species making up the total.

Animals↗
PubMed1985

Vascular permeability in experimental spinal cord injury.

Following spinal cord injury in rats there was a time-dependent change of vascular permeability as reflected by extravasation of 125I-labelled serum albumin. The change of vascular permeability correlated with tissue calcium and water accumulation suggesting that cord exposure to plasma calcium as a consequence of vascular injury may contribute to the progressive post-traumatic cord necrosis.

Animals↗
PubMed1985

Calcium-stimulated proteolysis in myelin: evidence for a Ca2+-activated neutral proteinase associated with purified myelin of rat CNS.

Incubation of myelin purified from rat spinal cord with CaCl2 (1-5 mM) in 10-50 mM Tris-HCl buffer at pH 7.6 containing 2 mM dithiothreitol resulted in the loss of both the large and small myelin basic proteins (MBPs), whereas incubation of myelin with Triton X-100 (0.25-0.5%) and 5 mM EGTA in the absence of calcium produced preferential extensive loss of proteolipid protein (PLP) relative to MBP. Inclusion of CaCl2 but not EGTA in the medium containing Triton X-100 enhanced degradation of both PLP and MBPs. The Ca2+-activated neutral proteinase (CANP) activity is inhibited by EGTA (5 mM) and partially inhibited by leupeptin and/or E-64c. CANP is active at pH 5.5-9.0, with the optimum at 7-8. The threshold of Ca2+ activation is approximately 100 microM. The 150K neurofilament protein (NFP) was progressively degraded when incubated with purified myelin in the presence of Ca2+. These results indicate that purified myelin is associated with and/or contains a CANP whose substrates include MBP, PLP, and 150K NFP. The degradation of PLP (trypsin-resistant) in the presence of detergent suggests either release of enzyme from membrane and/or structural alteration in the protein molecule rendering it accessible to proteolysis. The myelin-associated CANP may be important not only in the turnover of myelin proteins but also in myelin breakdown in brain diseases.

Animals↗
PubMed1985

Alteration of thromboxane and prostacyclin levels in experimental spinal cord injury.

We measured levels of thromboxane B2 and 6-keto-PGF1 alpha in rabbit spinal cord and cat CSF after impact injury to spinal cord. Rabbit tissue thromboxane B2 levels increased more than 6-keto-PGF1 alpha. In cat, CSF thromboxane B2 was higher the first hour postinjury; CSF 6-keto-PGF1 alpha also increased, but less so. These results imply activation of arachidonic acid metabolism. The relatively greater increase of thromboxane B2 suggests that thromboxane-prostacyclin imbalance may contribute to post-traumatic ischemia.

6-Ketoprostaglandin F1 alpha↗
PubMed1984

Tissue calcium levels in CaCl2-induced myelopathy.

CaCl2-induced myelopathy was produced in rats by the application of a solution of CaCl2 to exposed leptomeninges of lumbosacral spinal cord. Equal lengths of remote cervical and lumbar cord were removed at intervals following Ca2+ application. The total Ca2+ in tissue from lumbar cord was significantly elevated over autologous cervical cord and homologous lumber controls after after 2 h. The maximum Ca2+ increase in lumbar cord was 3.9-fold that of homogolous control and was reached by 8 h post Ca2+ application. The time course for the elevation of Ca2+ as measured by atomic absorption spectrophotometry, resembles that found in spinal cord following direct physical trauma. These findings as well as similar changes in morphology and neural proteins from previous studies suggest that Ca2+ is involved in and may potentiate the degeneration of axons and myelin via Ca2+-activated neutral proteinases.

Animals↗
PubMed1984

Glycosphingolipids of chicken skeletal muscle in early development and genetic dystrophy.

The acidic and neutral GSL of chicken pectoral muscle and the activities of relevant sialyltransferase and glycosidases have been examined during embryonic and early post-hatching development. At this stage of myogenesis, a prominent shift to the neutral GSL of longer oligosaccharide length involving Forssman glycolipid most prominently and also globoside and GbOse3Cer occurred but the distribution of muscle-type gangliosides was not obviously affected. The glycosidase and sialyltransferase activities decreased dramatically just prior to or at hatching. The fusion-linked change in GSL suggests a role for terminal galactosamine and/or galactose residues in myoblast aggregation. A parallel developmental study of genetic muscular dystrophy revealed similar GSL levels and enzyme activities. A larger proportion of lactosylceramide in dystrophic muscle throughout development suggests a developmental lag in the mutant.

Aging↗
PubMed1984

Glycosphingolipids of an alveolar rhabdomyosarcoma and normal human muscle. A case study.

The distribution of neutral, or asialosyl-, glycosphingolipids and gangliosides in a rhabdomyosarcoma of alveolar type have been studied. Histologically, this muscle tumor is composed primarily of two cell types: one with oval or round hyperchromatic nuclei and very little cytoplasm, and one a giant cell, with multiple, peripherally placed nuclei and weakly staining eosinophillic cytoplasm. In comparing glycolipids of the rhabdomyosarcoma with normal muscle from the same leg, the striking alteration in the tumor was a virtual disappearance of ganglioside GM2. There was also a slight increase in GM3 and a decrease in GD1a. The asialosyl derivative of GM2 (GalNac-Gal-Glc-Cer) was markedly increased in the tumor. A loss of glucosylceramide was also observed. The results are discussed in terms of glycolipid metabolic changes in muscle oncogenesis and their implications.

Child, Preschool↗
PubMed1984

A simplified procedure for the preparation of tritiated GM1 ganglioside and other glycosphingolipids.

The ganglioside II3NeuAc-GgOse4Cer and other glycosphingolipids can be radiolabeled to high specific activity by the galactose oxidase-NaB3H4 procedure, by purifying the oxidized compounds prior to reductive labeling. The oxidized products are separated from nonoxidized compounds and detergents (Triton X-100 and sodium taurocholate) present during the enzymatic oxidation. Since the oxidized derivatives are separated, the final specific activity depends solely upon the specific activity of the NaB3H4 and the reduction conditions.

Animals↗