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G D Armstrong

Publications and source records attributed to G D Armstrong.

At least 55 records · Page 3Linked to original sources
PubMed1983

N-terminal amino acid sequencing of EDP208 conjugative pili.

EDP208 conjugative pili contain a single polypeptide subunit of 11,500 daltons with a blocked N-terminus. This N-terminal blocking moiety was identified as an N-acetyl group by 1H nuclear magnetic resonance analysis of an N-terminal tripeptide isolated from pronase digests of EDP208 pilin. Limited acid hydrolysis of the tripeptide allowed its sequence to be determined as acetyl-NH-Thr-Asp-Leu. Trypsin digestion of EDP208 pilin resulted in the quantitative release of a fragment containing 12 residues from the N-terminus of the protein. The sequence of this dodecapeptide was determined to be acetyl-NH-Thr-Asp-Leu-Leu-Ala-Gly-Gly-Lys-Asp-Val-Asp-Lys.

Amino Acid Sequence↗
PubMed1983

Receptors for insulin and basic somatomedin: immunological and affinity-chromatographic cross-reactivity.

Human placental receptors for insulin and for human basic somatomedin (BSM, analogous to human insulinlike growth factor I (IGF-I] were selectively cross-link labelled with 125I-labelled insulin and BSM using disuccinimidyl suberate under conditions whereby cross-linking of 125I-labelled insulin to the BSM receptor and of 125I-labelled BSM to the insulin receptor was avoided. We have found that the cross-link-labelled receptors for insulin and BSM, present in equivalent amounts in human placenta membrane preparations, both cross-react with antibodies directed against purified rat liver insulin receptor. Compared with the human insulin receptor, the BSM receptor cross-reacted with the antireceptor antibody to a level of roughly 9%, indicating a limited sequence homology between the insulin and BSM receptors. Since both receptors are present in comparable amounts in solubilized placenta membrane preparations, we sought methods for the selective purification of both receptors from such extracts, so as to provide a basis for further comparative structural studies of the two receptors. We have observed that both receptors were adsorbed by affinity columns of insulin-agarose, in a manner that did not yield insulin receptor entirely free from the BSM receptor. As an alternative for receptor purification, we have found that immunoaffinity columns using antiligand antibody should provide a means for the selective isolation of cross-link labelled receptor from tissues in which both are present in equivalent amounts.

Chromatography, Affinity↗
PubMed1982

Comparative studies on human placental insulin and basic somatomedin receptors.

The disuccinimidyl suberate, affinity-labeling procedure, and proteolytic mapping techniques have been employed to characterize further the human placental receptors for insulin and basic somatomedin. Electrophoretic analysis of the basic somatomedin receptor, selectively crosslinked to 125I basic somatomedin in the presence of excess native insulin revealed, under reducing conditions, major labeled constituents of 270-280 and 125-140 kd, substantiating our previous work employing a photoaffinity labeling reagent. Affinity labeling also demonstrated the presence of less intensely labeled components with apparent molecular weights of 40 and 45 kd but failed to reveal a distinct 90- to 100-kd species observed in parallel experiments with insulin. In the absence of beta-mercaptoethanol, all components specifically labeled with 125I basic somatomedin migrated in the 300- to 400-kd range. In comparison, selective affinity labeling of the insulin receptor in the presence of excess native basic somatomedin revealed components, upon electrophoresis under reducing conditions, with apparent molecular weights of 270-280, 125-140, 90-100, and 40 kd. The major insulin-labeled component (125-140 kd) comigrated with the major constituent (125-140 kd) selectively labeled with basic somatomedin. When digestion was performed prior to solubilization, chymotryptic and tryptic proteolysis of the membrane-localized selectively labeled insulin, and basic somatomedin receptors yielded quite similar gel electrophoretic maps. However, when digestion was done subsequent to solubilization, chymotryptic and tryptic proteolysis of selectively labeled insulin and basic somatomedin receptors solubilized in SDS yielded similar but not identical gel electrophoretic maps. We conclude that the receptors for basic somatomedin and insulin are highly homologous structures with respect to their disulfide crosslinked composition, and with respect to the size of the major components detected by selective affinity-labeling procedures. Nevertheless, the detection of electrophoretically distinct labeled receptor components upon analysis of specifically labeled intact or proteolytically digested receptors points to subtle differences between the polypeptide compositions of the two receptors.

Disulfides↗
PubMed1982

Multidimensional assessment of psychological problems in children with cancer.

The Personality Inventory for Children (PIC) was administered to 88 children 3 to 16 years of age who had cancer. A variety of dimensions of psychological adjustment of these children, who were compared with a sample of nonsick children, was assessed. The PIC indicated that children with cancer exhibited cognitive development-related problems and internalized forms of psychopathology more often than their age- and sex-matched cohorts. In the cancer sample, no differences in problems as a function of diagnoses were observed when controlling for age of the child. No differences related to length of time since diagnosis were observed when controlling for age of the child. About 26% of the children with cancer exhibited PIC profiles indicating problems of such magnitude that professional intervention would be appropriate.

Adaptation, Psychological↗
PubMed1982

Characterization of the human placental receptor for basic somatomedin.

We have determined optimal conditions for the solubilization of the basic somatomedin (SM) receptor from human placental membranes and for the measurement of the binding of basic SM to the solubilized receptor. Further, we have developed conditions under which the basic SM receptor, in the presence of equivalent amounts of insulin receptor, can be selectively and specifically affinity-labeled with 125I-labeled basic SM, using the cross-linking reagent disuccinimidyl suberate (DSS). Our results with these developed methods indicate that the properties of the soluble basic SM receptor (pH optimum for ligand binding, pH 7 to 9; adsorption to lectin-agarose derivatives; sedimentation coefficient in detergent-sucrose solutions, 11S) closely parallel data previously reported for the insulin receptor. Based on the sedimentation coefficient and the previously estimated Stokes radius of the soluble receptor (7.2 nm), a molecular weight of 402 000 can be calculated for the detergent-receptor complex. Electrophoretic analysis of the basic SM receptor, selectively cross-linked to 125I-labeled basic SM with DSS in the presence of excess unlabeled insulin revealed, under reducing conditions, a major labeled constituent of 140 kdaltons, substantiating our previous work employing a photoaffinity labeling reagent. DSS cross-linking also demonstrated the presence of less intensely labeled components with apparent molecular weights of 54 000, 43 000 and 35 000 but failed to reveal a distinct 90- to 100-kdalton species visualized in parallel experiments with insulin. The 53-kdalton species was not detected in similar experiments with insulin. A specifically labeled basic SM receptor component of 300 kdaltons was also observed under reducing conditions; in the absence of beta-mercaptoethanol, all labeled components migrated in the 300-kdalton range. In comparison, selective DSS labeling of the insulin receptor in the presence of excess basic SM revealed components which, upon electrophoresis under reducing conditions, exhibited apparent molecular weights of 300 000, 140 000, 90 000--100 000, 43 000 and 35 000. The major insulin-labeled component (140 000) comigrated with the major constituent (140 000) selectively labeled with basic SM. Chymotryptic digestion of the receptors selectively DSS labeled with either 125I-labeled insulin or 125I-labeled basic SM yielded quite similar, but distinctive, gel electrophoretic maps. We conclude that the receptors for basic SM and insulin are highly homologous structures, particularly with respect to their glycoprotein nature, their hydrodynamic properties, their disulphide cross-linked composition, and with respect to the size of the major constituent detected by selective affinity labeling. Nonetheless, the detection of electrophoretically distinct labeled receptor substituents upon analysis of specifically labeled material, both before and after chymotryptic cleavage, points to subtle differences between the polypeptide compositions of the two receptors.

Female↗
PubMed1981

Nature of the carbohydrate and phosphate associated with ColB2 and EDP208 pilin.

Studies were carried out to elucidate the nature of phosphate and sugar linkages to F-like conjugative pili encoded by the ColB2Fdr (compatibility group FII) and EDP208 (compatibility group FV) plasmids. Both types of pili, when in the intact undissociated state, were found to contain approximately 3 mol of phosphate and 3 mol of sugar per mol of pilin. However, further purification of the two types of pilin by gel filtration chromatography in the presence of sodium dodecyl sulfate (SDS) removed all of the carbohydrate from EDP208 pilin and approximately 65% of the carbohydrate from ColB2 pilin. Approximately 0.8 to 1.0 mol of glucose per mol of protein remained associated with ColB2 pilin after SDS gel filtration chromatography, but it was not possible to determine whether this was covalently linked to the pilin, or tightly associated in an SDS-resistant manner. SDS-gel chromatography did not remove phosphate from either ColB2 or EDP208 pilins. (31)P nuclear magnetic resonance studies indicated that the pilin-associated phosphate is involved in a phosphodiester linkage. Acetone precipitation or chloroform-methanol extraction of the purified pilin material reduced the phosphate associated with EDP208 pilin to less than 0.04 molecule per pilin monomer. ColB2 pilin, under the same conditions, retained approximately 0.5 phosphate per pilin monomer. The extracted phosphate-containing moieties were identified as phosphatidylglycerol and phosphatidylethanolamine by thin-layer chromatography. Since the (31)P nuclear magnetic resonance spectra for both ColB2 and EDP208 were identical and no signal other than that of a phosphodiester was detected in the ColB2 spectrum, the phosphate remaining with the ColB2 pilin after chloroform-methanol extraction is most likely due to a tightly bound noncovalent residue.

Bacterial Proteins↗
PubMed1980

Comparative biochemical studies on F and EDP208 conjugative pili.

EDP208 pili are encoded by a derepressed derivative of a naturally occurring lac plasmid, F(0)lac (incompatibility group FV), originally isolated from Salmonella typhi. EDP208 pili are serologically unrelated to F pili and do not promote infection by F-specific ribonucleic acid bacteriophages. However, they do confer sensitivity to the F-specific filamentous deoxyribonucleic acid phages. EDP208-containing cells are multi-piliated and contain approximately 20 pili per cell. These pili contain a single polypeptide subunit of 11,500 daltons. EDP208-specific RNA phages were readily isolated from local sewage. These phages were somewhat smaller in diameter than the F-specific ribonucleic acid phages and absorbed relatively weakly to EDP208 pili. Comparing EDP208 pilin to F, it was found that both contain the equivalent of two to three hexose units per subunit as well as blocked N-termini. EDP208 pilin contains one covalently linked phosphate residue per subunit, whereas the F pilin subunit contains two such residues. Although notable differences were found in the case of three or four amino acids, the overall amino acid compositions of F and EDP208 were very similar. Moreover, the tryptic peptide maps of the two proteins contained seven peptides with similar mobilities, suggesting considerable homology in their amino acid sequences. Substantial similarities were also noted in the secondary structures of F and EDP208 pilin on the basis of circular dichroism studies. The alpha-helix content of both proteins was calculated to be 65 to 70%. X-ray fiber diffraction studies have indicated that the arrangements of subunits in F and EDP208 pili are also similar. It was concluded that F and EDP208 pili are closely related structures.

Bacterial Proteins↗
PubMed1979

Biochemical studies on pili isolated from Pseudomonas aeruginosa strain PAO.

Pseudomonas aeruginosa strains PAO and PAK bear polar pili which are flexible filaments having a diameter of 6 nm and an average length of 2500 nm. Both types of pili are retractile and promote infection by a number of bacteriophages. The present communication describes the partial biochemical characterization of PAO pili isolated from a multipiliated nonretractile mutant of PAO. The observed properties are compared to those of PAK pili which were characterized previously. PAO pili were found to contain a single polypeptide subunit of 18 700 daltons. This is similar to PAK pili which contain a single polypeptide of 18 100 daltons. The amino acid composition of PAO pilin was also similar to that of PAK pilin. Neither protein contained phosphate or carbohydrate residues and both were found to contain N-methylphenylalanine at the amino terminus. Sequencing of 20 amino acid residues at the amino terminal end of PAO pilin revealed the sequence to be identical with that of PAK pilin, while tryptic peptide analyses of PAO and PAK pilin indicated that the two proteins probably contain a number of homologous regions within the polypeptide. It was concluded that PAO and PAK pili were closely related structures.

Amino Acid Sequence↗
PubMed1977

Isolation and characterization of catabolite repression-resistant mutants of Escherichia coli.

A method has been developed for the isolation of Escherichia coli mutants which are resistant to catabolic repression. The method is based on the fact that a mixture of glucose and gluconate inhibits the development of chemotactic motility in the wild type, but not in the mutants. A motile E. coli strain was mutagenized and grown in glucose and gluconate. Mutants which were able to swim into a tube containing a chemotactic attractant (aspartic acid) were isolated. Most of these mutants were able to produce beta-galactosidase in the presence of glucose and gluconate and were normal in their ability to degrade adenosine 3',5-cyclic monophosphate. Some of these mutants were defective in the glucose phosphotransferase system.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMedDate not supplied

Immobilization of reducing sugars as toxin binding agents.

A simple and economical procedure for the attachment of reducing sugars to aminated solid supports has been developed. Reaction of the amino groups on the solid support with p-nitrophenyl chloroformate, followed by 1,6-hexanediamine, yields a chain-extended amine to which reducing sugars can be attached while remaining accessible to macromolecules. Immobilization of the reducing sugars involves a simple incubation followed by trapping of the resulting glycosylamine with acetic anhydride and recovery of the unreacted sugar by filtration. This technique was used to immobilize lactose and sialyllactose onto silylaminated Chromosorb P, producing solid supports that effectively neutralized the activity of cholera toxin from Vibrio cholerae and heat-labile enterotoxin of enterotoxigenic Escherichia coli. The general applicability of such solid supports for toxin neutralization was further demonstrated by immobilization of the enzymatically synthesized alpha Gal(1-3) beta Gal(1-4)Glc trisaccharide, which produced a support that efficiently neutralized toxin A of Clostridium difficile. The results from this study suggest that these solid supports have the potential to serve as inexpensive therapeutics for bacterial toxin-mediated diarrheal diseases.

Animals↗
PubMedDate not supplied

Vitamin ingestions.

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Adolescent↗
PubMedDate not supplied

Head nurse leadership style with staff nurse burnout and job satisfaction in neonatal intensive care units.

Leadership style has been defined as a two-factor construct composed of "consideration" and "initiating structure." Research has suggested that these factors affect the behavior and attitude of subordinates. This study's purpose was to quantify the relationships of head nurse leadership style with self-reported staff nurse burnout and job satisfaction in Neonatal Intensive Care Units (NICUs). Three instruments--the Minnesota Satisfaction Questionnaire, the Tedium Scale, and the Leadership Opinion Questionnaire--were voluntarily completed by 283 registered nurses employed by 14 level-III NICUs in the United States. The leadership dimensions of consideration and structure were distinct (r = -.10). Staff nurse satisfaction and burnout were related (r = -.41). Head nurse consideration was clearly related to staff nurse satisfaction (r = -.55) and to a lesser extent to burnout (r = -.29). Initiating structure alone was not related to satisfaction or burnout. Aggregate perceptions of head nurse leadership were ranked across NICUs in order to classify the head nurses on consideration and structure. The 14 head nurses were separated into four groups: high consideration-high structure, high consideration-low structure, low consideration-high structure, and low consideration-low structure. Satisfaction and burnout of staff nurses in each of the leadership-style groups were then compared. Analysis of variance for satisfaction (F(3,279) = 3.10, p = .03) and burnout (F(3,279) = 3.90, p = .01) were both significant. For both satisfaction and burnout, the head nurse leadership classification of low consideration-high structure was most deviant.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗