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G J Anderson

Publications and source records attributed to G J Anderson.

At least 91 records · Page 5Linked to original sources

Docosahexaenoic acid is the preferred dietary n-3 fatty acid for the development of the brain and retina.

The metabolism of individual dietary n-3 fatty acids was studied in n-3 fatty acid-deficient newly hatched chicks. Laying hens were fed the n-6 fatty acid, ethyl linoleate, as the only source of polyunsaturated fat. Chicks were then fed the n-3-deficient hens' diet, or one of three other diets supplemented with the ethyl ester of 18:3 n-3, 20:5 n-3 [eicosapentaenoic acid (EPA)], or 22:6 n-3 [docosahexaenoic acid (DHA)] at 0.44% of calories. At the end of 0, 1, 2, and 3 wk, the fatty acid composition of the brain, retina, liver, and serum was determined. Dietary EPA and DHA were equally effective at raising levels of DHA in the brain and retina. Dietary 18:3 was relatively ineffective in restoring brain and retina DHA. In the n-3-deficient chicks fed EPA or DHA, levels of DHA recovered to control values in both the brain and retina by 3 wk. Very little EPA accumulated in the brain or retina of chicks fed EPA. Hepatic synthesis of DHA from EPA appeared low, suggesting that the brain and retina synthesized the DHA that accumulated rapidly in these tissues after the feeding of EPA. The delta-4-desaturase enzyme was apparently very active, then, in the brain and retina. Retroconversion of dietary 22:6 to 22:5 and 20:5 was evident in the serum, liver, and retina but not in the brain. Thus, it was possible to study the relative metabolism and especially the interconversion of n-3 fatty acids in a environment uncomplicated by existing stores of these essential fatty acids. This study would suggest that 18:3 as the sole source of n-3 fatty acids in the diets of animals, including the human infant, may not be adequate for the biochemical development of the brain and retina and that dietary DHA is the preferred fatty acid of the n-3 series.

Animals↗

Conservation between yeast and man of a protein associated with U5 small nuclear ribonucleoprotein.

The process of nuclear pre-messenger RNA splicing is similar in Saccharomyces cerevisiae and metazoan cells in that the two-step mechanism is identical and the reaction occurs in a large ribonucleoprotein complex, the spliceosome. Little is known, however, about the degree of conservation of splicing factors other than of the small nuclear RNAs (snRNAs). Yeast counterparts of the metazoan spliceosomal snRNAs (U1, U2, U4, U5 and U6) have been identified but, with the exception of U6, the yeast snRNAs are larger and sequence similarity is limited to short regions. By using antibodies against the yeast PRP8 protein, a pre-mRNA splicing factor of relative molecular mass 280,000 (Mr280K) stably associated with U5 small nuclear ribonucleoproteins (snRNPs), we have now identified an immunologically related protein in HeLa cell nuclear extracts. The HeLa cell protein has an Mr greater than 200K and is associated with purified 20S U5 snRNPs. This is the first report of phylogenetic conservation between yeast and man of a protein splicing factor.

Autoantibodies↗

Methylscopolamine and conditioned location avoidance.

On alternate days, rats were confined to one side of a shuttlebox following IP administration of saline and to the other following the peripherally-acting muscarinic antagonist, methylscopolamine (1.2 mg/kg). They later avoided the side associated with the drug effect. By duplicating an earlier finding with centrally- and peripherally-acting scopolamine, this result identified aversive peripheral actions of the two drugs as mainly responsible for the effects observed.

Animals↗

Identification of H- and L-ferritin subunit binding sites on human T and B lymphoid cells.

Monoclonal antibodies to the H and L subunits of ferritin were used in indirect immunofluorescent studies to investigate the subunit specificity of ferritin binding to transformed T and B lymphoid cell lines. Pre-incubation of cells with recombinant H ferritin followed by antibodies to the H subunit produced a labelling pattern in which all cells showed strong fluorescent stippling. Corresponding experiments with liver ferritin and an antibody to the L subunit revealed that most cells were lightly stippled, but 3-5% of the cells were intensely labelled. These two labelling patterns were distinct as assessed by cross-blocking experiments with recombinant H and liver ferritins. These results imply that different binding sites exist for each subunit.

Antibodies, Monoclonal↗

Rapid modulation of the n-3 docosahexaenoic acid levels in the brain and retina of the newly hatched chick.

The newly hatched chick obtains its fatty acids almost completely from the lipids of the egg yolk as these are transferred to the developing embryo during its 21-day period of incubation. Since the diet of the laying hen greatly influences the fatty acid composition of the egg lipids, and presumably also the fatty acid composition of the resulting chick, we tested how quickly and to what extent varying the amount of n-3 fatty acids in the diet of the hen would modulate the level of n-3 fatty acids in the brain and retina of the newly hatched chick. White Leghorn hens were fed commercial or semi-purified diets supplemented with 10% fish oil, linseed oil, soy oil, or safflower oil. Eggs, together with the brain, retina, and serum of newly hatched chicks, were then analyzed for fatty acid composition. The fatty acids of egg yolk responded quickly to the hen's diet with most of the change occurring by 4 weeks. There was a linear relationship between the linolenic acid content of the diets and levels of this fatty acid in egg yolk and chick serum. In chicks from hens fed the fish oil diet, the total n-3 fatty acids, including 22:6(n-3), were elevated twofold in the brain and retina and sevenfold in serum relative to commercial diet controls. The safflower oil diet led to a very low n-3 fatty acid content in egg yolks and only 25% of the control n-3 fatty acid content in the brain and retina of chicks.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Nuclear pre-mRNA splicing in Saccharomyces cerevisiae.

While there are some differences in the nuclear pre-mRNA splicing machineries of Saccharomyces cerevisiae and higher eukaryotic cells, it is apparent that the fundamental mechanism of this reaction is highly conserved. S. cerevisiae is, therefore, an attractive organism for the study of splicing, since it is amenable to classical and molecular genetics as well as traditional biochemical methods. Here we present an outline of some of the advances which have resulted from this powerful combination of approaches.

Introns↗

Uptake of fatty acids by the developing rat brain.

Polyunsaturated fatty acids are avidly taken up by the developing rat brain. To explore the specificity of this process, [1-14C]labeled 16:0, 18:2n-6, 18:3n-3, and 22:6n-3 each were co-injected with [3H]18:1n-9 into the jugular vein of two-wk-old functionally hepatectomized and sham-operated control rats. The radioactivities present in the brain, liver and serum were assessed 30 min after injection. Uptake of labeled fatty acids into brain lipids steadily increased with increasing degree of unsaturation, with more than twice as much uptake of 22:6n-3 compared to 16:0. Phosphatidylcholine was the principal radioactive species in the brain except for animals injected with [1-14C]22:6n-3, in which more of the label was incorporated into phosphatidylethanolamine. Determination of water-soluble oxidation products in the brain and serum revealed that the greater uptake of the more unsatrated fatty acids did not result from differences in rates of degradation.

Animals↗

The anterior cricoid split: the Children's Hospital of Philadelphia experience.

The anterior cricoid split procedure (ACS) is an alternative to tracheotomy in selected infants with subglottic stenosis. From February 1983 to October 1987, 26 children underwent ACS at the Children's Hospital of Philadelphia. All children had subglottic stenosis. Six children had previously undergone tracheotomy. A successful outcome, allowing extubation or decannulation, correlated only with decreased pre-operative intubation time and was achieved in 73% of these patients. A failed ACS had little adverse affect on the subsequent management of these children.

Cricoid Cartilage↗

Identification of a yeast snRNP protein and detection of snRNP-snRNP interactions.

The RNA8 gene of Saccharomyces cerevisiae encodes an unusually large (260 kd) protein required for pre-mRNA splicing. Immunological procedures have been used to demonstrate that the RNA8 protein is in stable association with the small nuclear RNAs snR7L and snR7S, which are also known to be required for splicing and which are present in spliceosomal complexes. RNA8 is also involved in an ATP-dependent association with two other small nuclear RNAs, snR14 and snR6. It is proposed that this represents an ATP-dependent interaction between small nuclear ribonucleoprotein particles that precedes their entry into the spliceosome.

Adenosine Triphosphate↗

Transacylase activity of lactating bovine mammary fatty acid synthase.

An assay for the transacylation reaction catalyzed by fatty acid synthase was developed which does not require model substrates or labelled acyl-derivatives of CoA. It involves the transfer of the acyl group from unlabelled CoA to [3H]CoA. This assay shows the occurrence of transacylation at a relatively high rate with a variety of substrates that the enzyme is able to utilize. The activity is unaffected by dissociation of the enzyme or modification by iodoacetamide or 2-chloroacetyl-CoA.

Acetyltransferases↗

Desialylated transferrin as a serological marker of chronic excessive alcohol ingestion.

Partly desialylated transferrin was measured in the serum of subjects with chronic alcoholism, of patients with non-alcoholic-related steatohepatitis, diabetes, and other non-alcoholic liver diseases, and of healthy controls. In non-alcoholic patients and controls the maximum desialylated transferrin expressed in relation to total transferrin was 1.5%. This value was exceeded in 18 (90%) of the 20 alcoholics. By contrast, gamma-glutamyl transferase was within the reference range in 9 of the alcoholics.

Adult↗

Improved purification of the human placental transferrin receptor and a novel immunoradiometric assay for receptor protein.

A simplified method for the purification of human placental transferrin receptor is described. The procedure involves chromatography of a detergent extract of placenta on immobilized iron-loaded transferrin. Knowledge of the physiology of the interaction between transferrin and its receptor is applied to enable bound receptor to be eluted under mild conditions and essentially free of containing transferrin. Purified transferrin receptor and a monoclonal antibody to the receptor were used to develop a novel immunoradiometric assay for the receptor in which the monoclonal antibody is the radiolabelled species. A competition between two populations of receptor, one immobilized on a particulate support and the other in solution, provides the basis for the assay. Using this assay we have measured transferrin receptor levels in placental and hepatic tissue and in three cell lines during both the logarithmic and stationary phases of cell growth.

Antibodies, Monoclonal↗

Electrocardiographic diagnosis of posterior myocardial infarction revisited: a new approach using a multivariate discriminant analysis and thallium-201 myocardial scintigraphy.

This study examined the feasibility of using a multivariate discriminant analysis to design a useful electrocardiographic (ECG) model to diagnose posterior myocardial infarction (MI). Thallium-20) scintigraphy was used as a reference standard to identify posterior scar (fixed perfusion defects). The model was derived from 111 patients of whom 37 had fixed posterior defects and 74 had normal images, and its validity was subsequently tested in a separate group of 180 patients. In the initial group of patients, the fixed perfusion defects involved the posterior left ventricular wall alone in 15 patients, and the posterior and inferior walls in 22 patients. Stepwise multivariate discriminant analysis of 26 ECG variables produced a model of two variables (Q-wave duration in a VF and T-wave amplitude in V1) which provided a sensitivity of 78%, a specificity of 89%, and a predictive accuracy of 86% for the diagnosis of posterior MI. This model, when tested in the second group of 180 patients, yielded an overall prediction accuracy of 82% (sensitivity 65%, specificity 85%). Thus, the combination of Q-wave in a VF and upright T wave in V1 is the best ECG predictor of posterior MI. These two variables reflect the frequent association of posterior MI with inferior MI, and the reciprocal repolarization changes in the right precordial leads.

Adult↗

Simple radioimmunoassay for transferrin using insolubilized antitransferrin antibodies: its application to cultured cells.

A radioimmunoassay (RIA) for transferrin is described. The assay uses antitransferrin antibodies covalently coupled to the particulate support Matrex Pel 102, and is simple, sensitive, reproducible, and rapid. Transferrin measurement with this assay is independent of the degree of iron saturation of the protein. The RIA was applied to the measurement of transferrin concentrations in a variety of cultured human cells. Each of 11 cell lines studied contained endogenous transferrin, but the greatest concentration was found in Chang liver cells. Red blood cells were used as a negative control.

Antibodies↗

Fatty acid chain elongation by microsomal enzymes from the bovine meibomian gland.

The composition of meibomian gland lipids suggested that fatty acid chain elongation might play a major role in the synthesis of such lipids. A fatty acid synthase preparation from the bovine meibomian gland catalyzed the formation of C16 acid and the enzyme was immunologically quite similar to that in the mammary gland. The microsomal fraction from the gland, on the other hand, catalyzed elongation of endogenous fatty acids in the presence of ATP and Mg2+ and of exogenous C18-CoA using malonyl-CoA and NADPH as the preferred reductant. The elongated products, ranging up to C28 in chain length, were found mainly as CoA esters and products derived from them. With C18-CoA as the exogenous primer, the elongation rate was linear with incubation time up to 20 min but the rate changed in a sigmoidal manner with increasing protein concentration. The elongation rate was maximal at a pH around 7.0. Typical Michaelis-Menten-type substrate saturation patterns were observed with both malonyl-CoA and NADPH. From linear double-reciprocal plots, the Km values for the two substrates were calculated to be 52 and 11 microM, respectively, with a V of about 340 pmol min-1 mg protein-1 with respect to malonyl-CoA. Exogenous CoA esters of C16 to C22 fatty acids were elongated to give products up to C28 without exhibiting any preference for the primer. The present elongation system could account for the formation of most of the very long chains found in meibomian lipids.

Adenosine Triphosphate↗

Kinetic analysis of monocarboxylate uptake into perfused rat hearts.

To test a prediction that trans-sarcolemmal lactate movement is carrier mediated, 14C-labelled lactate or pyruvate and 3H-mannitol were transiently infused (2 min) into aerobically perfused rat hearts that had been depleted of intracellular monocarboxylates by a 30 min pre-perfusion in the absence of substrates. Uptake was calculated from the difference between the level of 14C-substrate predicted from the extracellular distribution of 3H-mannitol and the actual level (corrected for 14CO2 contamination) measured in successive 20 s samples of effluent perfusate. Computer optimization analysis of the initial influx revealed that monocarboxylate transport is not simply a question of diffusion. On the contrary, saturation and cross-inhibition (Ki, lactate 0.14 +/- 0.04 mM; Ki, pyruvate 2.2 +/- 0.4 mM; Ki, acetoacetate 6.2 +/- 0.9 mM; and Ki, 3-hydroxybutyrate 20 +/- 6.0 mM) suggest that trans-sarcolemmal monocarboxylate movements are reversibly mediated by a high activity (Vmax 34 +/- 7 mumol/min/g wet wt), low affinity lactate (Km 6 +/- 2 mM) permease. Further, studies into the effects of pH indicate that transport requires prior protonation of the carrier (pKa 7.1) and that lactate movements are driven by the displacement of the trans-sarcolemmal lactate and proton gradient from the Donnan equilibrium.

Animals↗