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Biomedical subjects

T Gill

Publications and source records attributed to T Gill.

At least 19 recordsLinked to original sources

Protamine-induced permeabilization of cell envelopes of gram-positive and gram-negative bacteria.

The inhibitory effect of the cationic peptide protamine on Listeria monocytogenes, Escherichia coli, and Shewanella putrefaciens has been studied in detail. The addition of protamine (10 to 1,000 micrograms/ml) resulted in inhibition of oxygen consumption after less than 1 min and loss of intracellular carboxyfluorescein and ATP after 2 to 5 min. Maximum antibacterial activity was reached at alkaline pH and in the absence of divalent cations. The efficient permeabilization of cell envelopes of both gram-positive and gram-negative bacteria suggests that protamine causes a general disruption of the cell envelope, leading to a rapid and nonspecific efflux of low- and high-molecular-weight compounds.

Adenosine Triphosphate

Functional characterization of the repeated UASINO element in the promoters of the INO1 and CHO2 genes of yeast.

In yeast, INO1 and CHO2 gene expression is subject to repression in response to inositol and choline supplementation. The response by both genes to inositol is controlled by a single set of regulatory factors and the highly conserved and repeated UASINO element (consensus: 5' CATGTGAAAT 3') that is found in multiple copies in both promoters. However, none of the native elements found in the INO1 and CHO2 promoters constitutes an exact match to the consensus element and the functionality of individual elements from these two promoters has not been tested. In this study, the function of individual putative UASINO elements from both promoters was tested by placing promoter fragments into a reporter construct which lacked a UAS element but contained the TATA element and start of transcription from the yeast CYC1 gene fused to the Escherichia coli lacZ gene. In addition, a set of oligonucleotides containing the consensus UASINO element with the first position systematically modified was also tested for UASINO function. These studies indicated that elements that contain a C or an A as the first base at the 5' end are functional to varying degrees. The majority of potential UASINO elements from the INO1 promoter were found to be inactive, whereas all of the elements from the CHO2 promoter tested were active. These results are discussed in light of the differential regulation of the two promoters.

Base Sequence

Changes in cell morphology of Listeria monocytogenes and Shewanella putrefaciens resulting from the action of protamine.

Protamine, which is an antibacterial basic peptide, was shown to alter the cell morphology of Listeria monocytogenes and Shewanella putrefaciens. Atomic force microscopy revealed that protamine smoothed the surface of cells, formed holes in the cell envelope, and caused fusion of S. putrefaciens cells. Immunoelectron microscopy of protamine-treated cells of both L. monocytogenes and S. putrefaciens showed great damage to the cell wall and condensation of the cytoplasm. Respiration of the cells was decreased due to treatment with sublethal concentrations of protamine, probably due to leakage or loss of cell envelope potential. It was concluded that protamine disrupted the outer surface structure and condensed the cytoplasm of sensitive cells and, in sublethal concentrations, altered membrane structures, thereby eliminating respiration.

Fluorescent Dyes

Antibacterial effect of protamine assayed by impedimetry.

Impedimetric measurements were used to assay the antibacterial effect of protamine. A good linear correlation between the impedance detection time and the initial cell counts was obtained (r = 0.99, n = 2). As basic peptides may cause clumping of cells, this correlation curve was used when estimating the cell number after protamine treatment, rather than colony counts. Protamine from salmon killed growing Gram-positive bacteria and significantly inhibited growth of Gram-negative bacteria in Tryptone Soy Broth (TSB) at 25 degrees C. In general Gram-positive bacteria were more sensitive to protamine than Gram-negative bacteria; the minimum inhibitory concentrations (MIC) determined for Gram-positive strains varied from 20 to 1000 micrograms ml-1 and for Gram-negative strains from 500 micrograms ml-1 to more than 4000 micrograms ml-1. The effect of protamine on non-growing Listeria monocytogenes Scott A suspended in buffer was not lethal as was the effect on growing cells; however, protamine (50-500 micrograms ml-1) killed the Gram-negative fish spoilage bacteria Shewanella putrefaciens when the live cells were suspended in buffer.

Animals

Revision total knee arthroplasty after failed unicompartmental knee arthroplasty or high tibial osteotomy.

A retrospective matched-pair comparative analysis was done between 30 total knee arthroplasties following failed high tibial osteotomies and 30 total knee arthroplasties following failed unicompartmental knee arthroplasties. The groups were matched according to age, gender, type of prosthesis, primary disease, and length of followup. A minimum followup of 2 years was required for inclusion in the study, and the average followup was 3.8 years (range, 2-9 years). The Knee Society Knee Score for the high tibial osteotomy group was significantly higher than that for the unicompartmental arthroplasty group. More osseous reconstructions were required in the unicompartmental revisions. Difficulty with exposure was not significantly greater in the osteotomy group. Rates of component loosening were not significantly different between the groups. A failed unicompartmental knee arthroplasty and a failed high tibial osteotomy can be revised successfully to a total knee arthroplasty. The results confirm that revisions after unicondylar arthroplasty and high tibial osteotomy are technically demanding. In this series, the results of total knee arthroplasty following unicompartmental knee arthroplasty approached but did not equal those obtained after high tibial osteotomy.

Aged

Measurement of the severity of amphetamine dependence.

A Severity of Amphetamine Dependence Questionnaire (SAmDQ) was administered to 101 subjects attending an Australian drug dependency treatment centre. The SAmDQ was adapted from the Severity of Opiate and Alcohol Dependence Questionnaires (SODQ & SADQ). The structural characteristics of the SAmDQ were examined and compared with previous findings reported on samples of opiate addicts with the SODQ. A high degree of consistency was found between the results obtained with the SAmDQ and previous findings with the SODQ. The relationship between the SAmDQ and the Severity of Dependence Scale (SDS) was also examined. The findings suggest that further development of amphetamine dependence measurement is required.

Adolescent

Regulation of phosphatidylethanolamine methyltransferase and phospholipid methyltransferase by phospholipid precursors in Saccharomyces cerevisiae.

Phosphatidylethanolamine methyltransferase (PEMT) and phospholipid methyltransferase (PLMT), which are encoded by the CHO2 and OPI3 genes, respectively, catalyze the three-step methylation of phosphatidylethanolamine to phosphatidylcholine in Saccharomyces cerevisiae. Regulation of PEMT and PLMT as well as CHO2 mRNA and OPI3 mRNA abundance was examined in S. cerevisiae cells supplemented with phospholipid precursors. The addition of choline to inositol-containing growth medium repressed the levels of CHO2 mRNA and OPI3 mRNA abundance in wild-type cells. The major effect on the levels of the CHO2 mRNA and OPI3 mRNA occurred in response to inositol. Regulation was also examined in cho2 and opi3 mutants, which are defective in PEMT and PLMT activities, respectively. These mutants can synthesize phosphatidylcholine when they are supplemented with choline by the CDP-choline-based pathway but they are not auxotrophic for choline. CHO2 mRNA and OPI3 mRNA were regulated by inositol plus choline in opi3 and cho2 mutants, respectively. However, there was no regulation in response to inositol when the mutants were not supplemented with choline. This analysis showed that the regulation of CHO2 mRNA and OPI3 mRNA abundance by inositol required phosphatidylcholine synthesis by the CDP-choline-based pathway. The regulation of CHO2 mRNA and OPI3 mRNA abundance generally correlated with the activities of PEMT and PLMT, respectively. CDP-diacylglycerol synthase and phosphatidylserine synthase, which are regulated by inositol in wild-type cells, were examined in the cho2 and opi3 mutants. Phosphatidylcholine synthesis was not required for the regulation of CDP-diacylglycerol synthase and phosphatidylserine synthase by inositol.

Cloning, Molecular

Assessment of right and left ventricular function in terms of force development with gestational age in the normal human fetus.

OBJECTIVE: To assess right and left ventricular systolic function in normal human fetuses by cross sectional Doppler echocardiography to calculate the force developed by myocardial shortening. DESIGN: Cross sectional echocardiographic images of the aorta and pulmonary arteries were obtained prospectively in order to measure great vessel diameters and calculate their cross sectional areas. Doppler velocity signals were recorded from the proximal aorta and the proximal pulmonary artery and digitised to obtain peak velocity, acceleration time, flow velocity time integral during acceleration, and the flow velocity time integral for the whole of ejection. Right and left ventricular force development was estimated by Newton's equation in which force is defined as the product of mass and acceleration. PATIENTS: 58 normal human fetuses at a gestational age of from 20 to 42 weeks. RESULTS: The cross sectional area of the pulmonary artery was 20% greater than that of the aorta. Aortic acceleration time was longer than that in the pulmonary artery, and peak blood flow velocity in the aorta was consistently greater than that in the pulmonary artery. Right ventricular stroke volume was significantly greater than left ventricular stroke volume. The force developed by the right and left ventricles was, however, similar throughout the gestational period studied, increasing tenfold from 20 weeks' gestation to term (r = 0.74, p less than 0.0001; r = 0.75, p less than 0.0001) respectively. CONCLUSION: The development of right and left ventricular force in the human fetus is similar in spite of the greater volume handled by the right ventricle. This index of ventricular performance does not require calculation of ventricular volume and because it varies independently of ventricular geometry and heart rate it should prove useful in assessing cardiac function in the normal human fetus and in fetuses with ventricular dysfunction.

Blood Flow Velocity

A vision for occupational therapy: tradition meets the future.

Advancing technology and world events will have an impact on the practice of occupational therapy. Computers, communication technology, and environmental concerns are all examples of issues that must be considered as major determinants in future practice decisions related to all groups. Incorporating these areas into the traditions of occupational therapy will result in a vital profession that cannot be replaced. However, these advances and events must not sway occupational therapists from their area of expertise. Technology will never be mastered if the profession continues to stray into areas of practice that do not form the core of occupational therapy.

Canada

Purification and characterization of purine nucleoside phosphorylase from Proteus vulgaris.

Purine nucleoside phosphorylase was isolated and purified from cell extracts of Proteus vulgaris recovered from spoiling cod fish (Gadus morhua). The molecular weight and isoelectric point of the enzyme were 120,000 +/- 2,000 and pH 6.8. The Michaelis constant for inosine as substrate was 3.9 x 10(-5). Guanosine also served as a substrate (Km = 2.9 x 10(-5). However, the enzyme was incapable of phosphorylizing adenosine. Adenosine proved to be useful as a competitive inhibitor and was used as a ligand for affinity chromatography of purine nucleoside phosphorylase following initial purification steps of gel filtration and ion-exchange chromatography.

Inosine

Muriel Driver Lecture 1986. You can't take it with you!

This paper addresses the issue of commitment, both to oneself through continuing education and to one's profession through involvement in provincial and national professional organizations. The results of a survey sent to 287 therapists are discussed in relation to a discussion of relevant literature. Results indicate that the respondents are quite involved in both areas with those between the ages of 26 and 40 being most involved in the professional associations. Personal satisfaction was the main reason for this involvement and time was the biggest deterrent to involvement.

Age Factors

A myelin basic protein-specific T lymphocyte line that mediates experimental autoimmune encephalomyelitis.

A T lymphocyte line, BP-1, expressing the T helper phenotype was selected from Lewis rats immunized with guinea pig myelin basic protein (GP-BP) in complete Freund's adjuvant (CFA). The BP-1 line responded specifically to GP-BP but not to PPD after the first round of selection, and responded to rat but not human or bovine BP. When injected i.p. into histocompatible Lewis or F1 (Lewis X P2) recipients, the BP-1 line induced both clinical signs of experimental autoimmune encephalomyelitis (EAE) and delayed type hypersensitivity (DTH) reactions in ears challenged intradermally with GP-BP but not PPD. The severity of clinical signs and the degree of ear swelling were dependent on the dose of BP-1 cells injected. Both activities were detectable with as few as 0.1 X 10(6) BP-1 line cells and required prior activation of the line cells with GP-BP presented by accessory cells. Lewis rats that had recovered from EAE induced by injection of GP-BP in CFA were more susceptible than naive rats to BP-1 line-mediated disease, requiring as few as 0.03 X 10(6) line cells. Clinical EAE and DTH could be serially transferred into F1 (Lewis X P2) recipients with BP-1 cells and back to nonirradiated Lewis parents with activated splenocytes, suggesting that BP-1 cells persist in recipient rats. These results demonstrate the potent biologic activities of an autoreactive BP-specific T lymphocyte line. This line possesses properties similar to BP lines described previously as well as to culture-conditioned splenic T effector cells; thus, the data presented here bridge the gap between these two approaches for studying T effector lymphocyte functions.

Animals

Cloning of eukaryotic genes in single-strand phage vectors: the human interferon genes.

Using oligonucleotide probes with defined sequences, we have selected clones from a human lymphocyte cDNA library which represent human leukocyte (HuIFN-alpha) and fibroblast (HuIFN-beta) interferon gene sequences. Double-stranded f1 phage DNA was used as the vector for initial cloning of cDNA. Clones carrying interferon gene sequences were identified by hybridization with the oligonucleotide probes. The same oligonucleotide probes were used as primers for dideoxy chain termination sequencing of the clones. One HuIFN-alpha clone, 201, has a nucleotide sequence different from published HuIFN-alpha sequences. Under control of the lacUV5 promoter, the 201 gene has been used to express biologically active HuIFN-alpha in Escherichia coli.

Base Sequence

Professional certification procedures: a developmental forecast for the year 2000.

This paper presents the technique of future forecasting through the development of a scenario based on the process of certification. The paper focuses on the demands of consumer groups. influence of world events, actions taken by Government, and developments within medicine, nursing and occupational therapy through the presentation of a scenario which spans the years 1983-2000. No claim is made that the contents of the scenario will become reality. Rather, the technique is presented as another tool that could be used by the profession in the development of future plans and policies.

Canada