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Biomedical subjects

R D Powell

Publications and source records attributed to R D Powell.

At least 19 recordsLinked to original sources
PubMed2001

Orbitally induced oscillations in the East Antarctic ice sheet at the Oligocene/Miocene boundary.

Between 34 and 15 million years (Myr) ago, when planetary temperatures were 3-4 degrees C warmer than at present and atmospheric CO2 concentrations were twice as high as today, the Antarctic ice sheets may have been unstable. Oxygen isotope records from deep-sea sediment cores suggest that during this time fluctuations in global temperatures and high-latitude continental ice volumes were influenced by orbital cycles. But it has hitherto not been possible to calibrate the inferred changes in ice volume with direct evidence for oscillations of the Antarctic ice sheets. Here we present sediment data from shallow marine cores in the western Ross Sea that exhibit well dated cyclic variations, and which link the extent of the East Antarctic ice sheet directly to orbital cycles during the Oligocene/Miocene transition (24.1-23.7 Myr ago). Three rapidly deposited glacimarine sequences are constrained to a period of less than 450 kyr by our age model, suggesting that orbital influences at the frequencies of obliquity (40 kyr) and eccentricity (125 kyr) controlled the oscillations of the ice margin at that time. An erosional hiatus covering 250 kyr provides direct evidence for a major episode of global cooling and ice-sheet expansion about 23.7 Myr ago, which had previously been inferred from oxygen isotope data (Mi1 event).

Journal Article↗
PubMed2000

New frontiers in gold labeling.

Recent advances in gold technology have led to probes with improved properties and performance for cell biologists: higher labeling density, better sensitivity, and greater penetration into tissues. Gold clusters, such as the 1.4-nm Nanogold, are gold compounds that can be covalently linked to Fab' antibody fragments, making small and stable probes. Silver enhancement then makes these small gold particles easily visible by EM, LM, and directly by eye. Another advance is the combination of fluorescent and gold probes for correlative microscopy. Chemical crosslinking of gold particles to many biologically active molecules has made possible many novel probes, such as gold-lipids, gold-Ni-NTA, and gold-ATP.

Gold↗
PubMed1999

Tetrairidium, a four-atom cluster, is readily visible as a density label in three-dimensional cryo-EM maps of proteins at 10-25 A resolution.

Heavy metal clusters derivatized to bind to designated chemical groups on proteins have great potential as density labels for cryo-electron microscopy. Smaller clusters offer higher resolution and penetrate more easily into sterically restricted sites, but are more difficult to detect. In this context, we have explored the potential of tetrairidium (Ir(4)) as a density label by attaching it via maleimide linkage to the C-terminus of the hepatitis B virus (HBV) capsid protein. Although the clusters are not visible in unprocessed cryo-electron micrographs, they are distinctly visible in three-dimensional density maps calculated from them, even at only partial occupancy. The Ir(4) label was clearly visualized in our maps at 11-14 A resolution of both size variants of the HBV capsid, thus confirming our previous localization of this site with undecagold (Zlotnick, A., Cheng, N., Stahl, S. J., Conway, J. F., Steven, A. C., and Wingfield, P. T., Proc. Natl. Acad. Sci. USA 94, 9556-9561, 1997). Ir(4) penetrated to the interior of intact capsids to label this site on their inner surface, unlike undecagold for which labelling was achieved only with dissociated dimers that were then reassembled into capsids. The Ir(4) cluster remained visible as the resolution of the maps was lowered progressively to approximately 25 A.

Capsid↗
PubMed1999

Metallosomes.

Structures and ordered arrays containing organometallic particles have potential application in nanofabrication, smaller computer components, optical devices, sensors, and membrane probes and as detection agents. Here, we describe construction of gold clusters covalently attached to lipids and their use in forming typical lipid structures: micelles, liposomes ("metallosomes"), and sheets on an air-water interface. Two sizes of gold clusters were used, undecagold, with an 11-gold atom core 0.8 nm in diameter, and the larger Nanogold, with a 1.4-nm gold core. The morphology of the structures formed was determined by electron microscopy at a resolution at which single gold-lipid molecules were visualized. Further modification by additional catalytic metal deposition enhanced detectability. The approach is flexible and permits a wide variety of metal particle structures to be created using known lipid structures as templates. Additionally, these gold-lipids may serve as useful membrane labels.

Calcium↗
PubMed1999

Ni-NTA-gold clusters target His-tagged proteins.

Addition of six histidines to recombinant proteins has proved useful in their purification by nickel-affinity columns. This technology was adapted by synthesizing the chelator for nickel (nitrilotriacetic acid, NTA) onto the surface of gold clusters. These Ni-NTA-gold clusters were shown to specifically target the 6His region of tagged proteins. Results were verified by column chromatography, dot and overlay blots, UV-Vis spectroscopy, and scanning transmission electron microscopy. A 6His-tagged adenovirus "knob" protein was also shown to maintain receptor binding activity after gold labeling. Two types of gold clusters were used: 1.4-nm Nanogold and a new 1.8-nm "PeptideGold" coated with an NTA-dipeptide-thiol. These novel labels should be useful in site-specific high-resolution EM labeling, as well as in metallographic development, detection in the light microscope, or direct visualization.

Adenoviridae↗
PubMed1999

Giant platinum clusters: 2-nm covalent metal cluster labels.

A new class of covalently linkable platinum cluster reagents with core diameters close to 2 nm has been prepared. The new label offers the performance advantages and versatility of the 1.4-nm Nanogold in a larger label which is more clearly visualized against electron-dense regions of a specimen. Large platinum clusters were prepared by the reduction of platinum(II) acetate in the presence of 1,10-phenanthroline ligands which had been synthetically modified to include solubilizing and reactive cross-linkable functional groups. These were then conjugated site-specifically to antibody IgG molecules and to Fab' fragments and visualized by scanning transmission electron microscopy. The resulting conjugates may be autometallographically enhanced and show sensitivity similar to that of Nanogold conjugates in immunoblotting experiments. In preliminary experiments, they have also exhibited labeling of tissue antigens and penetrate to access nuclear targets.

Affinity Labels↗
PubMed1998

Combined fluorescent and gold immunoprobes: reagents and methods for correlative light and electron microscopy.

Immunoprobes which incorporate both a fluorescent label and a 1.4 nm gold cluster compound were prepared by covalent conjugation to Fab' antibody fragments of the Nanogold cluster label followed by a fluorescent moiety. These new immunoconjugates allow the collection of two complementary sets of data, from fluorescence and electron microscopy, from a single labeling experiment. By using Fab' fragments, the entire probe is smaller than a whole IgG molecule. A simple fluorescence assay was used to investigate the fluorescence properties of the new probes. They were used to localize the pre-mRNA splicing factor SC35 in the HeLa cell nucleus by both fluorescence and electron microscopy, and also for labeling leukocyte microtubules; labeling was imaged using fluorescence microscopy and, after silver enhancement, by a variety of optical methods and by electron microscopy. Combined Nanogold and Texas Red, Cy3, Lissamine Rhodamine B, and AMCA probes were also prepared, and in preliminary experiments show similar properties to the combined fluorescein and gold cluster probes. The fluorescent and gold cluster probes enable a new degree of correlation between fluorescence and electron microscopy, and may also be used to check labeling of specimens before processing for electron microscopy.

Cell Nucleus↗
PubMed1997

A covalent fluorescent-gold immunoprobe: simultaneous detection of a pre-mRNA splicing factor by light and electron microscopy.

Immunoprobes that combine a fluorescent label with a 1.4-nm gold cluster compound have been prepared by covalent conjugation with polyclonal antibody Fab' fragments. These new immunoconjugates allow the collection of two complementary sets of data, from fluorescence and electron microscopy, from a single labeling experiment. We find that incorporation of one or more fluorescein moieties into the coordinated ligands of the 1.4-nm Nanogold gold cluster label yields a stable, dual-function immunolabel in which fluorescence quenching is negligible. In a second synthetic strategy, Nanogold and fluorescein were separately covalently conjugated to Fab' fragments to yield a probe with very similar properties. With the use of Fab' fragments, the entire probe is smaller than a whole IgG molecule, and it exhibited excellent penetration properties. It was used to localize the pre-mRNA splicing factor SC35 in the HeLa cell nucleus by both fluorescence and electron microscopy.

Fluorescein↗
PubMed1993

Orgasm consistency training in the treatment of women reporting hypoactive sexual desire: an outcome comparison of women-only groups and couples-only groups.

To evaluate and compare the effectiveness and maintenance of two group interventions using orgasm consistency training in the treatment of female hypoactive sexual desire, 57 women were randomly assigned to a women-only group, a couples-only group, or a waiting list control group. Controlling for social desirability, subjects were assessed on six variables: sexual compatibility, sexual esteem, sexual desire, sexual fantasy, sexual assertiveness, and sexual satisfaction. Independent assessments were made on these variables before treatment, after treatment, and at 6 months follow-up. Although the treatment was found to be generally effective in women reporting hypoactive sexual desire, a consistent pattern of change favoring the couples-only group was evident on all measures. Possible explanations for the superiority of couples-only interventions are explored in the discussion.

Adult↗
PubMed1991

Standard-setting plans for the NBME comprehensive Part I and Part II examinations.

The National Board of Medical Examiners (NBME) has reviewed its procedure for setting pass-fail standards in conjunction with the introduction of its comprehensive Part I and Part II examinations in 1991. This report gives background information on the procedures used for the past decade to set pass-fail standards for the Part I and Part II examinations, an overview of the NBME's research on standard setting, under way since 1987, and a statement of its plans for determining pass-fail standards for these examinations. In 1981 the NBME changed from the norm-referenced standard, used since the 1950s, to a criterion-group approach to setting pass-fail standards. Although the criterion-group system resulted in more stable standards, it still meant that the standard moved whenever the performance of the reference group changed. After conducting research, surveying constituencies, and examining alternatives, the NBME has adopted a new standard-setting plan that has the following components: a content-based standard-setting procedure; determination of standards by an appropriate group; use of a fixed standard; and periodic review of standards and standard-setting procedures. This new process will produce three types of improvements: it will incorporate deliberations informed by a wide range of information, including content review; annual review of examinees' performances and pass-fail results and triennial restudy of the process will add further quality control; and a fixed standard will mean that comparable performances will be required across administrations in order to pass.

Education, Medical, Undergraduate↗
PubMed1985

Scintigraphic findings in primary amyloidosis of the heart and stomach.

The effectiveness of scintigraphic studies in the study of patients with primary systemic amyloidosis has not been reported previously. This case report describes a patient with the scintigraphic findings of thickened myocardium and delayed gastric emptying, which correlated well with the presence of amyloid deposits in these organs at autopsy. Additionally, the liver and spleen had normal scintigraphic appearances and were free of amyloid at autopsy. Radionuclide studies appear useful in assessing the anatomic and physiologic abnormalities assessed by amyloid deposition.

Aged↗
PubMed1984

Retention of intact cells on bronchoscopy brushes after cleaning.

Reusable nylon bristle bronchoscopy brushes were evaluated to assess whether cytologically recognizable cells could be retained on the bristles after conventional cleaning techniques. Twenty brushes were used during bronchoscopy; 5 brushes were inserted directly into freshly resected neoplasms. The brushes were then scrubbed and rinsed in chlorhexidine gluconate, iodine, and alcohol, and air dried. Several days later fresh cytologic specimens were prepared from the apparently clean brushes. Eighty percent of the cytologic specimens exhibited strands of mucus, most containing well-preserved alveolar macrophages, polymorphonuclear leukocytes, and ciliated epithelial cells. Although no malignant cells were retained, it is clear that conventional cleaning techniques are inadequate for the removal of cells and acellular debris. Consideration should be given to immersing brushes in acetylcysteine or employing disposable brushes.

Bronchoscopy↗
PubMed1981

High-temperature Disruption of Guard Cells of Vicia faba: EFFECT ON STOMATAL APERTURE.

Increased variability in stomatal aperture at high temperatures can be attributed, in part, to the differential sensitivity of guard cells to thermal damage. Individual stomata become increasingly open at higher temperatures until guard cells are lethally damaged; at that temperature, apertures decrease. The extent of irreversible damage causing closure was estimated by K(+) uptake, neutral red accumulation, and visual scoring of chloroplasts.This study found that visual scoring of chloroplast disruption provided the best estimate of guard cell viability at high temperature. Removal of the damaged guard cells from the population sample resulted in a constant coefficient of variability for apertures over the temperature range 25 to 50 C.

Journal Article↗