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Biomedical subjects

D Noonan

Publications and source records attributed to D Noonan.

At least 55 records · Page 3Linked to original sources

Campylobacter jejuni/Campylobacter coli-associated Guillain-Barré syndrome. Immunoblot confirmation of the serological response.

Immunoblot (Western blot) examination of the sera of 45 patients with the Guillain-Barré syndrome demonstrated that between three and five immunoreactive bands that were characteristic of a recent Campylobacter jejuni/Campylobacter coli infection were present in 22 of these patients. The immunoblots paralleled the serological response that is found in campylobacter enteritis and confirmed the specificity of our previous enzyme-linked immunosorbent assays (ELISAs), which indicates that antecedent C. jejuni/C. coli infection is common in patients with the Guillain-Barré syndrome. We postulate that, under certain circumstances, demyelination is initiated by the leakage of campylobacter-specific antibody across the blood-nerve barrier.

Campylobacter Infections↗

A library of trimethylguanosine-capped small RNAs in Physarum polycephalum.

We have constructed a cDNA library for the trimethylguanosine-capped small RNAs (sRNAs) in the acellular slime mold Physarum polycephalum. Capped sRNAs were purified from total cellular RNA of vegetative microplasmodia by preparative immunoprecipitation with anti-trimethylguanosine antibody. The purified RNA was analyzed by polyacrylamide gel electrophoresis. Approx. eleven different capped sRNAs were observed with a size range of 70-204 nucleotides (nt). Based on their approximate sizes, the presence of trimethylguanosine cap, and the presence of a lupus type-Sm antigen, molecules U1-U7 (excluding U3) were identified. Further confirmation of the identity of molecule U1a was established by Northern hybridization, U4a by colony hybridization, and U6 and U7a by direct chemical sequence analysis. Purified capped sRNAs were tailed with oligo(A), and inserted into oligo(dT)-tailed plasmid pCDV1. The cDNAs were used to transform Escherichia coli strain HB101. Approx. 1.9 X 10(5) ampicillin-resistant (ApR) transformants were obtained per microgram of tailed sRNA. Dot-blot hybridization, using Physarum RNA precipitated with anti-cap antibody as a probe, indicated that approx. 94% of the ApR colonies contained recombinant DNAs. The library was screened by colony hybridization using heterologous sRNA probes. Clones hybridizing with heterologous sRNAs U1, U2, U4 and U7 were each represented in the library in approximately the same frequency as their relative abundance in the Physarum sRNA population they were derived from. The insert of one Physarum U4 clone was sequenced and was found to have 57.1% homology with nt 1-91 of the published sequence for rat U4 RNA. A 12-nt 'functional' subdomain of the rat U4 molecule was 83.3% conserved in Physarum U4.

Autoantigens↗

Cytoplasmic processing events in the polyadenylate region of Physarum messenger RNA.

Cytoplasmic processing events in the poly(A) region of mRNA from Physarum polycephalum are reviewed. Two classes of poly-containing RNA [poly(A)+ RNA] exist in the cytoplasm. One contains very short poly(A) sequences, averaging about 15 adenylate residues, while the other contains relatively long poly(A) sequences, averaging about 60 residues. Molecules with short poly(A) sequences are found exclusively in the polysomes while those with long poly(A) sequences are restricted to the free cytoplasmic mRNP. Since proteins are associated with only the long poly(A) sequences the poly(A) . protein complex is also restricted to the free mRNP. The long poly(A) sequences are relatively short-lived. They are degraded by two distinct processes, a shortening process in which 15-20 residues are gradually removed and a turnover process in which long poly(A) tracts are rapidly converted to the short sequences. This process, along with the dissociation of the poly(A) . protein complex, occurs when poly(A)+ RNA molecules located in free mRNP are transferred to the polysomes. Poly(A) . protein complex dissociation appears to precede poly(A) turnover during translational selection. The significance of these processing events in relation to mRNA maturation is discussed.

Cell Nucleus↗

Cytoplasmic polyadenylate processing events accompany the transfer of mRNA from the free mRNP particles to the polysomes in Physarum.

The relationship between the mRNA in the polysomes and the free cytoplasmic messenger ribonucleoprotein of Physarum polycephalum was studied by microinjection techniques. Labeled free cytoplasmic ribonucleoprotein, prepared from donor plasmodia, was microinjected into unlabeled host plasmodia, and its fat was followed in the host ribonucleoprotein particles. Approximately one-half of the poly(A)-containing RNA [poly(A)+RNA] that originated from the microinjected particles was incorporated into the host polysomes by normal translational processes within 1 hr. Very short poly(A) sequences (approximately 15 nucleotide residues) were found in these poly(A)+RNA molecules. These short poly(A) sequences were sensitive to digestion with micrococcal nuclease, suggesting that they were not associated with protein. Because the poly(A)+RNA molecules of the microinjected free cytoplasmic mRNP had originally contained poly(A) sequences 50-65 nucleotides long and were associated with protein extensive poly(A) degradation and poly(A).protein complex dissociation must have occurred during their incorporation into the polysomes or during their translation. These results demonstrate a precursor-product relationship between free cytoplasmic mRNP and polysomal mRNA and suggest that the incorporation process in Physarum is accompanied by structural modifications in the poly(A) region of mRNA. They also imply that the polysome is a site for disruption of the poly(A).protein complex and poly(A) degradation.

Cytoplasm↗

Stored messenger ribonucleoprotein particles in differentiated sclerotia of Physarum polycephalum.

Starvation induces vegetative microplasmodia of Physarum polycephalum to differentiate into translationally-dormant sclerotia. The existence and the biochemical nature of stored mRNA in sclerotia is examined in this report. The sclerotia contain about 50% of the poly (A)-containing RNA [poly(A)+RNA] complement of microplasmodia as determined by [3H]-poly(U) hybridization. The sclerotial poly(A)+RNA sequences are associated with proteins in a ribonucleoprotein complex [poly(A)+mRNP] which sediments more slowly than the polysomes. Sclerotial poly(A)+RNP sediments more rapidly than poly(A)+RNP derived from the polysomes of microplasmodia despite the occurrence of poly(A)+RNA molecules of a similar size in both particles suggesting the existence of differences in protein composition. Isolation of poly(A)+RNP by oligo (dT)-cellulose chromatography and the analysis of its associated proteins by polyacrylamide gel electrophoresis show that sclerotial poly(A)+RNP contains at least 14 major polypeptides, 11 of which are different in electrophoretic mobility from the polypeptides found in polysomal poly(A)+RNP. Three of the sclerotial poly(A)+RNP polypeptides are associated with the poly(A) sequence (18, 46, and 52 x 10(3) mol. wt. components), while the remaining eight are presumably bound to non-poly(A) portions of the poly(A)+RNA. Although distinct from polysomal poly(A)+RNP, the sclerotial poly(A)+RNP is similar in sedimentation behavior and protein composition (with two exceptions) to the microplasmodial free cytoplasmic poly(A)+RNP. The results suggest that dormant sclerotia store mRNA sequences in association with a distinct set of proteins and that these proteins are similar to those associated with the free cytoplasmic poly(A)+RNP of vegetative plasmodia.

Base Sequence↗

The poly(adenylic acid)-protein complex is restricted to the nonpolysomal messenger ribonucleoprotein of Physarum polycephalum.

The distribution of poly(adenylic acid) [poly(A)]-protein complexes in the polysomal and nonpolysomal messenger ribonucleoprotein (mRNP) fractions of Physarum polycephalum was examined in the present study. Poly-(A)-containing components released from the nonpolysomal mRNP by ribonuclease (RNase) digestion were quantitatively adsorbed to nitrocellulose filters at low ionic strength, were highly resistant to micrococcal nuclease under conditions in which free poly(A) was completely degraded, and sedimented as a 10-15S particle which was disrupted by sodium dodecyl sulfate and protease treatment. These are characteristics of the poly(A)-protein complex. In contrast,poly(A)-containing molecules released from the polysomes by RNase were refractive to nitrocellulose, were completely sensitive to micrococcal nuclease, and sedimented at 2-4 S, identical with the sedimentation exhibited by protein-free poly(A). Examination of the poly(A) sequences present in polysomal and nonpolysomal mRNP by polyacylamide gel electrophoresis showed that the former contained only very short sequences, averaging approximately 15 nucleotides, while the latter exhibited only much longer segments, averaging approximately 65 nucleotides. It is concluded that poly(A)-protein complexes are restricted to the nonpolysomal mRNP of Physarum and that the limiting factor in complex formation may be the length of the available poly(A) binding site.

Molecular Weight↗

A comparison of methods of blood withdrawal and sample preparation for potassium measurements.

The effect on measured potassium values of a number of factors involved in sample acquisition and preparation were investigated. Measured potassium values in either plasma or serum were not influenced by the site of sample withdrawal, the presence of a tourniquet, or the time elapsed between blood sampling and analysis. However, increasing heparin concentrations and/or volumes decreased potassium values. The highest values were obtained in samples obtained in a syringe that had been washed with minimum volumes of heparin. It is concluded that this common clinical practice is based on a sound scientific rationale.

Arteries↗

KSHV sequences in biopsies and cultured spindle cells of epidemic, iatrogenic and Mediterranean forms of Kaposi's sarcoma.

The pathogenesis of Kaposi's sarcoma (KS) is still unclear, and several factors appear to be involved in the onset of the Kaposi's lesion. Epidemiological studies suggest that a common infective agent may contribute to KS. Sequences which appear to represent a new gammaherpesvirus, currently termed KSHV/HHV8, have recently been identified in KS. To further examine the relationship between this virus and KS, we obtained biopsy samples of KS lesions; these samples, the spindle cells cultured from these lesions and the PBMC of the same patients were tested for the presence of KSHV sequences by PCR. In addition, we tested several "late passage" KS spindle cell lines as well as control samples. The biopsy samples were from lesions of the following forms of KS: one sporadic KS, two epidemic KS and three iatrogenic KS, one of which was in the process of regressing after reduction of immunosuppressive therapy, and two that were at different stages (patch and nodular) from a single patient. The sporadic KS specimen was positive, as were the PBMCs from this patient, and cells grown from this biopsy appeared to contain KSHV viral sequences up to the fifth passage. Both epidemic KS biopsies were positive, but in these cases KSHV sequences were not detected in the cultured cells. The biopsy from the regressing iatrogenic KS lesion was negative, as were the cells cultured from this lesion. However, the PBMCs of this patient were weakly positive for KSHV at the time of biopsy, and PBMCs collected from this patient one month later were completely negative. The samples of both the patch and the nodular KS lesions obtained from another immunosuppressed patient showed amplifiable sequences of KSHV, but both the PBMCs of this patient and primary KS cell cultures from these biopsies were negative. Of the late-passage KS lines tested, only one, IST AIDS KS 12, was positive for KSHV. This line is derived from an early angiomatous-macula lesion. Taken together, these data suggest that an active KSHV infection is associated with KS and that elimination of KSHV from the lesion precedes regression of the lesion, strongly correlating KSHV with KS. In addition, early KS lesions may have a higher KSHV burden, or contain cells more susceptible to KSHV infection, further linking KSHV to KS.

Aged↗

Shaping healthcare: developing a program evaluation questionnaire.

Healthcare delivery today emphasizes quality improvement in program management. The focus on consumer needs and satisfaction challenges us to develop effective tools to evaluate our services from the user's perspective. This article delineates the development, implementation, analysis, and outcomes of a program evaluation questionnaire that was designed to evaluate program staff and treatment services from the perspectives of both patients and families. The potential this type of consumer opinion gauge and tool has to shape healthcare delivery is outlined within the context of a rehabilitation program.

Aged↗

Angiogenesis at the interface between basic and clinical research.

The field of antiangiogenesis has shown a remarkably rapid evolution from the discovery at the bench to translation into the clinic. Currently a wide variety of compounds are in clinical trial as inhibitors of angiogenesis, and new compounds are being frequently added. The target cell of most angiogenesis inhibitors is the endothelial cell, with inhibitors that selectively affect a number of endothelial cell functions acquired during angiogenesis, including activation, proliferation, migration, invasion and survival. The endothelial cell may also be targeted by chemotherapeutic agents currently in use. The high doses and intermittent treatment schedules used to fight resistant tumor cells may be altered towards lower doses and chronic administration to obtain selective inhibition of angiogenic factor-stimulated endothelial cells as adjuvant therapy. Finally, gene therapy is a promising route for the delivery of novel protein inhibitors of angiogenesis, and is actively being investigated.

Angiogenesis Inducing Agents↗

Generation of expression plasmids for angiostatin, endostatin and TIMP-2 for cancer gene therapy.

Antiangiogenic therapy may represent a promising approach to cancer treatment. Indeed, the efficacy of endogenous angiogenesis inhibitors, including angiostatin, endostatin and TIMPs, has been demonstrated in many types of solid tumors in animal models. In view of the possible problems associated with long-term administration of inhibitors as recombinant proteins, we propose their delivery as nucleic acids through a gene therapy approach. To this end, eukaryotic expression constructs for murine angiostatin and endostatin as well as human TIMP-2 were generated, and characterized in vitro. All constructs carry the relevant cDNAs under the control of the strong HCMV promoter/enhancer, and cleavable leader signals to allow protein secretion. Expression of the angiogenesis inhibitors was detected by in vitro transcription/translation experiments as well as transfection of 293T cells, followed by Western blotting (WB) or radioimmunoprecipitation analysis of both cell lysates and supernatants (SNs). These constructs might be used for in vivo intramuscular delivery of plasmid DNA and as a set of reagents for the development of retroviral as well as adeno-associated viral (AAV) vectors expressing angiogenesis inhibitors.

Angiogenesis Inhibitors↗

Immunoblot confirmation of immune response to Campylobacter pyloridis in patients with duodenal ulcers.

The specificity of serum immune responses to Campylobacter pyloridis that were found in the sera of patients with duodenal ulcers has been confirmed by means of the Western blot method. The presence of the same immune response to the organism in apparently healthy individuals, especially older people, suggests a repeated exposure throughout life to this organism, the source of which remains unknown.

Adult↗

Rethinking social work service delivery.

Providence Centre is a 577-bed, multilevel, geriatric health facility located in Scarborough, Ontario. This article outlines the transition from a traditional model of social work service delivery to a specialized case referral system. It describes the review process, creation of the alternative service model, implementation strategies, evaluation measures and outcomes of the undertaking. Through these efforts, the centre succeeded in creatively redefining and restructuring its social work service delivery and resource utilization in today's tough economic climate.

Aged↗

Participative management and shared leadership: implementing a model.

The author identifies the development, implementation and outcomes of a task subgroup model of management that provides a mechanism for shared leadership, planning, decision making, implementation and evaluation by staff, patients and families on a program level. The conceptual model and its operationalization are outlined within the context of the rehabilitation program at the Providence Centre in Scarborough, Ontario.

Decision Making, Organizational↗