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Biomedical subjects

D Fisher

Publications and source records attributed to D Fisher.

At least 19 recordsLinked to original sources
PubMed1995

NH2-terminal truncation of skeletal muscle troponin T does not alter the Ca2+ sensitivity of thin filament assembly.

To investigate how Ca2+ binding to troponin C regulates muscle contraction, the Ca(2+)-sensitive properties of thin filament assembly were studied as the tropomyosin binding, NH2-terminal region of troponin T was progressively shortened. Troponin complexes were prepared that contained skeletal muscle troponin C, troponin I, and either intact troponin T (TnT) (residues 1-259) or fragment TnT-(70-259), TnT-(151-259), or TnT-(159-259). In the absence of Ca2+ their respective affinities for pyrene-labeled tropomyosin were 2.3 x 10(7) M-1, 1.2 x 10(7) M-1, 1.9 x 10(5) M-1, and 1.9 x 10(5) M-1. Ca2+ had only a small effect on these affinities: 1.1 x 10(7) M-1 for whole troponin, 2 x 10(5) M-1 for troponin-(151-259), and 2.8 x 10(5) M-1 for troponin-(159-259). Forms of troponin that bound weakly to tropomyosin in the absence of actin increased the actin affinity of tropomyosin only 2-3-fold, even in the absence of Ca2+; weak binding of troponin to tropomyosin correlated with weak effects on tropomyosin-actin binding. In contrast, whole troponin had an approximately 500-fold effect on tropomyosin binding to actin, regardless of whether Ca2+ was present. The small effect of Ca2+ on the energetics of thin filament assembly is not attributable to the amino-terminal region of troponin T. The results suggest that Ca2+ causes the interaction between actin and the globular region of troponin to switch between two energetically similar states.

Actin Cytoskeleton↗
PubMed1995

Oxalosis of bone: report of four cases and a new radiological staging.

Oxalosis of bone is caused by a combination of crystalline calcium oxalate deposition in bone and renal osteodystrophy in patients with chronic renal failure. The present paper presents the radiographic findings of oxalosis of bone in four patients who have been followed over a period of from 4 to 16 years. Four successive characteristic stages can be recognized in the development of the skeletal changes.

Adolescent↗
PubMed1995

Cyclins of the fission yeast Schizosaccharomyces pombe.

Five cyclin-like genes, cig1, cig2/cyc17, mcs2, puc1 and cdc13, have been discovered in S. pombe to date. It is not yet clear what their functions are or even whether they are all involved with control of the cell cycle. Conflicting data for cig1 and cig2/cyc17 have obscured analysis of their function and cig1 remains largely uncharacterized, although clues to the role of cig2/cyc17 have emerged. There is genetic data available for the more distant cyclin homologue mcs2, which has an essential although as yet unspecified role. Puc1 may be involved in regulation of exit from the cell cycle. The first cyclin to be discovered, and the best understood, is cdc13 which with cdc2 promotes mitosis. Studies of the roles of cdc2 and cdc13 in the overall ordering of the cell cycle suggest that cdc13 and probably other cyclins are key regulators, maintaining the order of S phase and mitosis during the cell cycle.

Cell Cycle↗
PubMed1995

Disseminated Penicillium marneffei: presenting illness of advanced HIV infection; a clinicopathological review, illustrated by a case report.

BACKGROUND: Until recently Penicillium marneffei rarely caused human disease. It is now a frequently encountered opportunistic mycosis in HIV positive residents of, and travellers to, south-east Asia. AIMS: To review aspects of clinical presentation, pathology, treatment, epidemiology and ecology of P. marneffei. To report a case of disseminated P. marneffei occurring in the Northern Territory which illustrates many typical clinical and pathological features of this infection. CASE PRESENTATION: A Burmese immigrant presented to Royal Darwin Hospital, Australia with a non-specific, subacute, febrile illness and a diffuse papular rash. The etiological agent was Penicillium marneffei, disseminated in association with advanced HIV infection. The typical travel history and umbilicated papular rash were recognized on admission. Fungal stains of skin biopsies and touch smears facilitated rapid diagnosis, and early antifungal therapy resulted in clinical cure. CONCLUSIONS: Early distinction of penicilliosis from other opportunistic mycoses, tuberculosis, Leishmaniasis, and molluscum contagiosum is critical for effective management. The characteristic histological and mycological properties of P. marneffei are easily recognizable if the diagnosis is considered. In view of geographic proximity, travel and immigration from endemic areas, Australia should expect further imported penicilliosis as illustrated by this case report.

AIDS-Related Opportunistic Infections↗
PubMed1995

The pharmacokinetics and neuromuscular effects of rocuronium bromide in patients with liver disease.

To determine the effect of liver disease on the pharmacokinetics of rocuronium, the authors administered 0.6 mg/kg (twice the ED95) to 10 patients with liver disease and compared these results to values in 10 healthy surgical patients. Anesthesia was induced with thiopental and maintained with isoflurane (0.9%-1.1% end-tidal concentration) and nitrous oxide (60%). Venous blood samples were obtained for 6 h after rocuronium injection and plasma concentrations were measured using gas chromatography. Pharmacokinetic differences between groups were determined using a population-based pharmacokinetic analysis (NONMEM). Hepatic impairment did not alter the plasma clearance of rocuronium (217 +/- 21.8 mL/min, mean +/- SE, for both groups), but did increase the volume of the central compartment (5.96 +/- 1.01 L for controls, 7.87 +/- 1.33 L for patients with liver disease) and volume of distribution at steady state (16.4 L for controls, 23.4 L for patients with liver disease). In turn, elimination half-life was longer in patients with liver disease (111 min) compared to controls (75.4 min). The authors conclude that liver disease alters the pharmacokinetics of rocuronium by increasing its volume of distribution. The longer elimination half-life might result in a longer duration of action of rocuronium in patients with liver disease, particularly after prolonged administration.

Adult↗
PubMed1994

Evaluation of an automated DNA profiling system employing multiplex amplification of four tetrameric STR loci.

We have examined the performance and reproducibility of an automated DNA profiling system which is based on the multiplex amplification of 4 tetrameric STR loci-HUMVWFA31/A. HUMTH01, HUMF13A1 and HUMFES/FPS. The system was able to type 100 pg of purified, undegraded, genomic DNA. At lower concentrations of DNA (below 100 pg), allelec drop-out occurred due to stochastic differences in allele copy number. Minor variation of individual PCR reagent concentrations or cycling temperatures did not result in a significant effect on the efficiency of amplification of any of the 4 loci in the quadruplex system. More substantial variation of reagent concentrations or cycling temperatures outside the optimum range of the system resulted in a reduction or complete loss of signal for one or more loci. This was also observed at high ionic strength or extreme pH. However, under all reagent concentrations and conditions studied, no artefact bands that could potentially result in the mistyping of a sample were apparent within the read region (130-240 bases) of the gel. Evaluation of both native and denaturing polyacrylamide gels revealed that, although native gels displayed faster run times, the sizing precision of such gels for certain STR loci was lower than that of denaturing gels. Also, artefact bands may be present within the read region of native gels. In conclusion the quadruplex amplification system described, coupled with automated fluorescence-based detection on denaturing polyacrylamide gels, appeared to be a robust and reliable system for individual identification.

Alleles↗
PubMed1994

Separation of alkaline phosphatase isoforms with and without intact glycan-phosphatidylinositol anchors in aqueous polymer phase systems.

Alkaline phosphatase (ALP) isoforms can be distinguished from each other by their partition characteristics in aqueous two-phase systems composed of water-soluble polymers, the phases of which are differentially sensitive to the presence of glycan-phosphatidylinositol anchors. Compared with detergent-based systems, the aqueous polymer systems have the advantage that micelle formation does not take place. Partition of anchor-intact and anchor-degraded molecules in the latter systems is further improved by attachment of a hydrophobic ligand to one of the phase forming polymers. In this way, anchor-intact molecules can be separated from molecules with degraded anchors in a single partition step. The method has been used to confirm that ALP in human serum is predominantly anchor degraded, whereas in bile it retains its anchor intact.

Alkaline Phosphatase↗
PubMed1994

Quantitative analysis of polyethylene glycol (PEG) in PEG-modified proteins/cytokines by aqueous two-phase systems.

Covalent attachment of poly(ethylene glycol) (PEG) to proteins produces conjugates with altered/improved physicochemical and biological properties which depend upon the number of PEG chains linked. Quantification of the attached PEG is however not a trivial issue. The partition coefficient, K, of the PEG-protein conjugate in PEG/dextran two-phase systems provides a quantitative measure for the degree of modification. A linear relationship between log K and the number of PEG chains was observed in fractionated PEG-modified-granulocyte-macrophage colony stimulating factor conjugates having 1 to 3 substitutions. Furthermore, in mixtures of PEG-bovine-serum-albumin conjugates with increasing degrees of modification, a linear relationship was found between log K and n, the average substitution. The increment in log K per PEG chain added is protein specific and this suggests that the interactions between the PEG-protein conjugate and the polymers in the phase system are more complex than just a simple affinity of the PEG for the PEG-rich top phase. Increasing the polymer concentration in the phase system produces larger increments in log K per PEG molecule attached and the proportionality between log K and number of PEG molecules is only compromised for conjugates with high degree of substitution when partitioned in biphasic systems of high concentration of polymers.

Chemical Phenomena↗
PubMed1993

Radiolabeled-antibody therapy of B-cell lymphoma with autologous bone marrow support.

BACKGROUND: Radiolabeled monoclonal antibodies recognizing B-lymphocyte surface antigens represent a potentially effective new therapy for lymphomas. We assessed the biodistribution, toxicity, and efficacy of anti-CD20 (B1 and 1F5) and anti-CD37 (MB-1) antibodies labeled with iodine-131 in 43 patients with B-cell lymphoma in relapse. METHODS: Sequential biodistribution studies were performed with escalating doses of antibody (0.5, 2.5, and 10 mg per kilogram of body weight) trace-labeled with 5 to 10 mCi of 131I. The doses of radiation absorbed by tumors and normal organs were estimated by serial gamma-camera imaging and tumor biopsies. Patients whose tumors were estimated to receive greater doses of radiation than the liver, lungs, or kidneys (i.e., patients with a favorable biodistribution) were eligible for therapeutic infusion of 131I-labeled antibodies according to a phase 1 dose-escalation protocol. RESULTS: Twenty-four patients had a favorable biodistribution, and 19 received therapeutic infusions of 234 to 777 mCi of 131I-labeled antibodies (58 to 1168 mg) followed by autologous marrow reinfusion, resulting in complete remission in 16, a partial response in 2, and a minor response (25 to 50 percent regression of tumor) in 1. Nine patients have remained in continuous complete remission for 3 to 53 months. Toxic effects included myelosuppression, nausea, infections, and two episodes of cardiopulmonary toxicity, and were moderate in patients treated with doses of 131I-labeled antibodies that delivered less than 27.25 Gy to normal organs. CONCLUSIONS: High-dose radioimmunotherapy with 131I-labeled antibodies is associated with a high response rate in patients with B-cell lymphoma in whom antibody biodistribution is favorable.

Antibodies, Monoclonal↗
PubMed1993

Phase partitioning detects differences between phospholipase-released forms of alkaline phosphatase--a GPI-linked protein.

A number of enzymes are known to release alkaline phosphatase and other glycan phosphatidylinositol-anchored proteins from membrane surfaces. We describe a novel approach to detect and measure these activities by partitioning in aqueous phase systems. The procedures avoid the complications of micelle-formation involving hydrophobic molecules that may arise with detergent-based partition systems and can clearly distinguish between inositol-specific phospholipase C and D activities.

Alkaline Phosphatase↗
PubMed1992

Nuclear magnetic relaxation dispersion and 31P-NMR studies of the effect of covalent modification of membrane surfaces with poly(ethylene glycol).

Covalent attachment of methoxypoly(ethylene glycol) (MPEG) 5000 to the surface of unilamellar liposomes composed of egg phosphatidylcholine and dioleoylphosphatidylethanolamine (DOPE) (8:2) containing paramagnetic chelates, either entrapped within the interior volume of the liposomes, or associated with the membrane surface, had no effect upon the measured spin-lattice relaxation rates (1/T1) for water in these systems. 31P-NMR studies indicate no destabilization of dioleoylphosphatidylcholine (DOPC)/(DOPE) (1:1) vesicles following attachment of MPEG. However, in DOPC/DOPE (1:3) mixtures, covalent modification with MPEG results in a destabilization of multilamellar vesicles into smaller vesicular structures. These results indicate that covalent attachment of poly(ethylene glycol) to liposomal magnetic resonance agents may prove a useful method for increasing their utility as vascular MR agents by extending their lifetime in the circulation, without decreasing the relaxivity of paramagnetic species associated with the liposome, but that the presence of PEG covalently attached to the membrane surface may modify the polymorphic phase behavior of the lipid system to which it is covalently linked.

Lipid Bilayers↗
PubMed1992

Erythrocyte partitioning in dextran-poly(ethylene glycol) aqueous phase systems. Events in phase and cell separation.

Early events in the partitioning process which involve characteristic kinetics of cell- and phase-specific interactions and phase separation have been described previously. This paper reports on red cell-phase droplet interactions pertaining at the time of usual phase sampling (i.e., the time at which a clear bulk interface is first apparent) and beyond in cell partitioning and countercurrent distribution experiments. In non-charge-sensitive phase systems close to the critical point, cells can be free or attached to phase droplets. Cells that are free are virtually completely in the top phase, whereas different cell populations that show essentially complete binding to droplets can nevertheless have different partition ratios and be separated, thus reflecting the effects of the difference in the cells' avidity for the phase droplets during the early, elapsed events in partitioning. At higher polymer concentrations (i.e., higher interfacial tensions), the cell populations, completely bound to phase droplets, partition completely to the interface, and consequently cannot be separated. When such systems are made charge-sensitive by the generation of a Donnan potential between the phases or made into affinity systems by the incorporation of PEG ligands (e.g., PEG-palmitate), there is a decrease in the avidity of the cells for phase droplets. The resulting increase in the ratio of free to droplet-bound red cells in the top phase at the time of sampling correlates with an increase in the partition ratio, P, observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Heterogeneity in the circulating neutrophil pool: studies on subpopulations separated by continuous flow electrophoresis.

Isolated blood neutrophils from healthy individuals have been separated by continuous flow electrophoresis (CFE) as a Gaussian-shaped profile extending over 12-15 fractions, on the basis of differences in cell surface electrical charge. The fractions were pooled into three or four subpopulations; the mean electrophoretic mobilities of the least and most electronegative cells were 0.96 and 1.22 microns/sec/V/cm, respectively. Each pool of neutrophils was analyzed for functional and biochemical differences. Expression of respiratory burst in terms of the rate of superoxide production by the least and most electronegative cells to fixed concentrations of N-formyl-methionyl-leucyl-phenylalanine (fMLP, 10(-7) M) or phorbol myristate acetate (PMA, 1 microgram/ml) revealed that the least electronegative cells generated superoxide anion (O2-) at approximately twice the rate of the most electronegative cells. However, when lower concentrations of fMLP were used (1-5 x 10(-9) M), the most electronegative cells were most active. The least electronegative cells were also the most active in terms of phagocytosis and chemotaxis. In accordance with these differences in motile function, the basal F-actin content of the least electronegative cell pool was greater than the F-actin levels found in the most electronegative cells and remained so upon stimulation with fMLP. Such neutrophil heterogeneity as detected by CFE may be important in selective margination and recruitment of cells to inflammatory foci and sites of infection and may in part represent subsets of cells within the circulation that are primed in vivo to respond to inflammatory stimuli.

Actins↗