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Biomedical subjects

R A White

Publications and source records attributed to R A White.

At least 19 recordsLinked to original sources
PubMed1992

Changing patterns in cytoskeletal mRNA expression and protein synthesis during murine erythropoiesis in vivo.

The major cytoskeletal proteins alpha-spectrin, beta-spectrin, and ankyrin are synthesized and assembled into a supportive membrane skeleton during erythroid differentiation. Information on the temporal appearance of mRNA and protein species is essential for understanding both the cytoskeletal assembly process and the function of various isoforms. We have isolated highly enriched populations of fetal erythroid cells at various stages of maturation. mRNAs for erythroid ankyrin, alpha-spectrin, and beta-spectrin were expressed at all stages but there were differences in transcript types and levels. The ratio of 9-kilobase (kb) to 7.5-kb erythroid ankyrin transcripts decreased markedly during differentiation, but there was no change in the ratio of the 10.1-kb and 9.3-kb erythroid beta-spectrin transcripts. The relative amounts of ankyrin, alpha-spectrin, and beta-spectrin mRNA increased during yolk sac cell differentiation, whereas only alpha-spectrin mRNA increased during differentiation of the fetal liver cells. The amounts of beta-spectrin mRNA exceeded the amounts of alpha-spectrin mRNA in the early precursors from both yolk sac and fetal liver; protein synthetic levels showed the same pattern. The 16-day fetal peripheral reticulocytes, on the other hand, had the adult mRNA and protein synthetic ratios with alpha/beta greater than 1. The data indicate that at least two mechanisms exist to meet changing erythroid membrane cytoskeletal requirements during development in utero: (i) stage-specific processing of the mRNA for the major cytoskeletal linker protein ankyrin and (ii) developmentally regulated alpha/beta-spectrin protein synthetic rates.

Animals↗
PubMed1992

A quantitative method for evaluating bivariate flow cytometric data obtained using monoclonal antibodies to bromodeoxyuridine.

A method is presented for analyzing data from bivariate analysis of cell populations exposed to bromodeoxyuridine and subsequently examined both for the presence of BrdUrd and for the cellular DNA content. It is shown that certain features may be defined in the bivariate data which are constant independent both of cell type and, within limits, experimental variability. These landmark features include the ratio of red, DNA, fluorescence of G2 + M cells to G1 cells, the ratio of green fluorescence corresponding to the non-specific binding of unlabeled G2 + M cells to unlabeled G1 cells, and the distribution of green fluorescence in unlabeled cells. The landmarks make it possible to standardize rules for establishing the separation line between-labeled and unlabeled cells as required in these experiments to obtain estimates of cytokinetic parameters. Values obtained for the DNA synthesis time and the potential doubling time which result from different decision rules for distinguishing labeled from unlabeled are compared in two murine tumor lines. The potential doubling time, but not the DNA synthesis time is shown to depend sensitively on the separation line. Suggestions are presented for analyzing clinical data with this procedure.

Animals↗
PubMed1992

On the measurement of cytokinetics by continuous labeling with bromodeoxyuridine with applications to chick wing buds.

The cytokinetic properties, specifically the phase-transit times, TG1, TS, and TG2+M, of chick wing bud cells were estimated using data obtained from continuous labeling of stage 20 embryos with bromodeoxyuridine (BrdUrd). The presence of BrdUrd was detected with monoclonal antibodies, and the amount of DNA in the cells was determined with propidium iodide. The fraction of cells in each cell cycle phase, the fraction of labeled cells, and the relative movement, a measure of the mean DNA content, of all labeled cells were evaluated using bivariate flow cytometry at successive times following introduction of the label. Equations are presented to describe the fraction of unlabeled cells in G2 + M, which gives a direct estimate of TG2+M; the fraction of all labeled cells, which can then be used to estimate TG1; and, finally, the relative movement, which provides an estimate of TS. Thus, the data measured in these experiments together provide estimates of the progression through the cell cycle of limb mesoderm cells.

Animals↗
PubMed1992

Holmium:YAG laser ablation of human intervertebral disc: preliminary evaluation.

Percutaneous discetomy has become a viable alternative in the treatment of herniated intervertebral disc. This study determined the effectiveness of holmium: YAG laser for ablation of human disc tissue. Human cadaveric intervertebral disc was harvested and stored in cold saline-soaked gauze for evaluation within 24 hr of removal. Using a specially designed apparatus, a 600 microns diameter fiber was advanced perpendicular through the annulus fibrosis at a controlled force of 0.098 Newtons (10 g). Samples were lased in air (n = 17) and in room temperature saline (n = 32). The laser energy was delivered at 5 Hz, 250 microseconds pulsewidth, and from 50 mJ/mm2 to 1,100 mJ/mm2 fluence. Three to six holes were lased using identical parameters in each tissue specimen and were evaluated histologically and by morphometric analysis. The maximum zone of thermal necrosis and thermal denaturation occurred at 700-1,100 mJ/mm2; 140 microns and 590 microns in air and 80 microns and 730 microns in saline, respectively. At fluences between 200 and 700 mJ/mm2, the thermal necrosis ranged from 20 to 60 microns in air and from 10 to 50 microns in saline, the zone of denaturation also being less. The holes created with the 600 microns fiber were circular in shape, with a mean diameter of 500 microns (n = 3). The etch rates (penetration/pulse) appeared to increase with increasing fluences. In saline, the etch rate ranged from 7 to 53 microns/pulse (r = 0.57, P less than or equal to 0.10), and, in air, the values ranged from 7 to 65 microns/pulse (r = 0.79, P less than or equal to 0.03).(ABSTRACT TRUNCATED AT 250 WORDS)

Acoustics↗
PubMed1992

Intravascular ultrasound guided holmium:YAG laser recanalization of occluded arteries.

Current angioplasty devices are limited by arterial wall dissection and perforation, and by early recurrence from inadequate debulking of lesions. This study evaluated intravascular ultrasound (IVUS) as guidance for concentric laser recanalization of arterial occlusions. Twelve, 2-4-cm-long canine iliac artery occlusions were treated at 2 weeks (organizing thrombus) to 12 weeks (firm fibrous lesions) using a Holmium:YAG laser (2,100 nm wavelength) in free running mode, FRM, (250 musec pulse, 5 Hz), n = 9; and Q-switched mode, QSM (200 ns pulse, 6 Hz), n = 3. A 200 microns (n = 2) or 600 microns (n = 10) optic fiber was centered in the artery coaxial to a 5Fr rotating A scan IVUS probe. The fiber was positioned in the center of the artery distal to the lesion and slowly advanced through the obstruction. In 8 occlusions the same fiber was used as a guidewire for passage of either a 1.6-mm-(n = 2) and/or 3.0-mm (n = 6) diameter multifiber catheter (19 x 100 and 19 x 200 microns fibers, respectively) using FRM energy to further debulk the lesion. In all cases, IVUS guidance enabled concentric initial recanalization of occlusions, although 3 vessel perforations resulted from fiber deviation off the center of the lumen at a distance of 2 to 4 cm from the IVUS imaging element. Both QSM and FRM modes ablated tissue, with FRM modes producing more tissue fragmentation and thermal effect. IVUS images accurately diagnosed the location of lesions compared to angioscopic views and pathologic analysis of the specimens.(ABSTRACT TRUNCATED AT 250 WORDS)

Angioplasty, Laser↗
PubMed1992

Murine erythrocyte ankyrin cDNA: highly conserved regions of the regulatory domain.

Ankyrin is an essential link between cytoskeletal proteins, such as spectrin, and membrane bound proteins, such as protein 3, the erythrocyte anion exchanger. Although the amino acid structure of human ankyrin is known, the functional regions have been only partially defined. Sequence comparisons between mouse and human ankyrin offer one mechanism of identifying highly conserved regions that probably have functional significance. We report the isolation and sequencing of a series of overlapping murine erythroid ankyrin (Ank-1) cDNAs from spleen and reticulocyte libraries (total span 6238 bp) and identify potentially important regions of murine-human reticulocyte ankyrin homology. Comparison of the predicted peptide sequences of mouse and human erythroid ankyrins shows that these ankyrins are highly conserved in both the N-terminal, protein 3 binding domain (96% amino acid identity) and in the central spectrin-binding domain (97% identity), but differ in the C-terminal regulatory domain (79% identity). However, the C-terminal regulatory domain contains two regions of peptide sequence that are perfectly conserved. We postulate these regions are important in the regulatory functions of this domain.

Amino Acid Sequence↗
PubMed1992

Sequential intraluminal ultrasound evaluation of balloon angioplasty of an iliac artery lesion.

This report describes intravascular ultrasound imaging of a localized 81% stenosis in the left common iliac artery of a 52-year-old woman. The lesion was dilated using an 8 mm balloon and was imaged pre- and post-dilation using arteriography and a 5F, 30 Mhz intravascular ultrasound catheter. The same site was imaged again intraoperatively at two months following the initial procedure using an 8F, 20 Mhz intravascular ultrasound catheter when the patient had a femoropopliteal bypass for continuing ischemia. Intravascular ultrasound imaging allowed accurate, sequential, on-line calculation of the cross-sectional area and volume of the lesion both, acutely and following healing of the site. The morphology of the fractured arterial plaque was clearly defined, demonstrating distribution of calcification, and intraluminal flaps not apparent on arteriography. The case demonstrates the unique potential of intravascular ultrasound in assessing the immediate effect of interventions and evaluating the long-term healing.

Angioplasty, Balloon↗
PubMed1992

Intravascular ultrasonography.

Intravascular ultrasonography is developing rapidly as a method for defining the transmural anatomy of vascular structures, with diagnostic and therapeutic applications. The ultrasound technology not only has unique diagnostic capabilities by defining the distribution and character of lesions, but also provides accurate control information regarding efficacy of angioplasty methods. An exciting recent development is the three-dimensional reconstruction of two-dimensional images which permits global examination of luminal and transmural vessel morphology. This technology may enable improved guidance of intraluminal devices to enhance lesion removal without damaging adjacent normal wall structure and appropriate device selection by differentiating specific plaque characteristics.

Animals↗
PubMed1992

The murine pallid mutation is a platelet storage pool disease associated with the protein 4.2 (pallidin) gene.

Pallid is one of 12 independent murine mutations with a prolonged bleeding time that are models for human platelet storage pool deficiencies in which several intracellular organelles are abnormal. We have mapped the murine gene for protein 4.2 (Epb4.2) to chromosome 2 where it co-localizes with pallid. Southern blot analyses suggest that pallid is a mutation in the Epb4.2 gene. Northern blot analyses demonstrate a smaller than normal Epb4.2 transcript in affected pallid tissues, such as kidney and skin. This is the first gene defect to be associated with a platelet storage pool deficiency, and may allow the identification of a novel structure or biological pathway that influences granulogenesis.

Animals↗
PubMed1992

Three-dimensional transrectal ultrasound: preliminary patient evaluation.

Transrectal ultrasound is uniquely useful in diagnosing and localizing the extent of lower colon and rectal disease. This paper reports the preliminary evaluation of three-dimensional intraluminal ultrasound imaging of normal and diseased rectal segments. The three-dimensional reconstructions were produced using a computerized PC based image analysis system which aligns ultrasound images to produce the three-dimensional images. A unique perspective for displaying both normal and pathologic anatomy is achieved using this new technology and the method has promising diagnostic and therapeutic potentials.

Crohn Disease↗
PubMed1992

Connectin, a target of homeotic gene control in Drosophila.

The homeotic genes of Drosophila encode transcription factors that specify morphological differences between segments. To identify the genes that they control, we developed a chromatin immunopurification approach designed to isolate in vivo binding sites for the products of the homeotic gene Ultrabithorax. Here, we report the analysis of one immunopurified binding site. This 110 bp fragment maps within a regulatory region of a gene under homeotic control, connectin. A 4 kb DNA fragment, including the immunopurified binding site, is sufficient to reproduce the appropriate homeotic control within a subset of the full tissue distribution of connectin. Analysis of the role of the 110 bp binding site indicates that it mediates transcriptional controls by Ultrabithorax and other homeotic genes. This is the first report of a functional in vivo binding site isolated using the chromatin immunopurification method. We also show that the protein product of the connectin gene is predicted to be a cell-surface molecule containing leucine-rich repeats. The protein, connectin, can mediate cell-cell adhesion thus suggesting a direct link between homeotic gene function and processes of cell-cell recognition.

Amino Acid Sequence↗
PubMed1992

The 412 retrotransposon and the development of gonadal mesoderm in Drosophila.

We have shown that the expression of the 412 retrotransposon provides a useful early marker for the development of the gonadal mesoderm in Drosophila embryos. 412 is initially expressed in a set of parasegmentally repeated stripes from parasegments (PS) 2-14 in the mesoderm at the extended germ band stage. During germ band retraction the bulk of 412 expression declines except in dorsolateral clusters of cells in PS10, 11 and 12, where high levels of 412 expression remain. These mesodermal cell clusters are associated with germ cells and subsequently they coalesce, rounding up to form the gonads. The gonadal mesoderm thus appears to originate specifically from three abdominal parasegments, PS10, 11 and 12. We show that the maintenance of high levels of 412 expression in gonadal mesoderm is not induced by contact with germ cells, but rather depends on genetic control by the homeotic genes abdominal-A and Abdominal-B.

Animals↗
PubMed1992

Targets of homeotic gene regulation in Drosophila.

We have used a chromatin immunopurification approach to identify target genes regulated by the homeotic gene Ultrabithorax. A monoclonal antibody against the Ultrabithorax gene product is used to immunopurify in vivo Ultrabithorax protein binding sites in embryonic chromatin. The procedure gives an enrichment of sequences with matches to a consensus homeodomain binding site. In one case we have shown that an immunopurified sequence lies within a 4 kb fragment that acts in vivo as a homeotic response element. We anticipate that this approach will enable us to identify further targets, allowing the analysis of their regulation and function. The chromatin immunopurification strategy may be of general application for the identification of direct in vivo targets of DNA-binding proteins.

Animals↗
PubMed1992

Three-dimensional intravascular ultrasound imaging of normal and diseased canine and human arteries.

This study reports three-dimensional reconstruction of two-dimensional intravascular ultrasound images obtained along 5 cm vessel segments. Each three-dimensional image was produced by computerized "stacking" of a set of consecutive two-dimensional images (mode 90 images per set; range 32 to 256). Three-dimensional images (n = 26) were obtained from 11 human normal and atherosclerotic arteries (three in vitro and eight in vivo) and five in vivo canine studies. In vivo human examinations included three iliac, one deep, and three superficial femoral arteries and one aortic dissection. Five in vivo canine vessels (three iliac stenoses and two aortic dissections) were imaged before and after intraluminal stent deployment. Images were displayed on a gray-scale monitor, allowing examination of vessel images as complete cylinders or longitudinal hemisections in any user-defined plane. This enabled global examination of vascular segments and identified luminal shape, wall thickness, and distribution and morphology of plaques. Reconstructions of aortic dissections illustrated the extent of the dissection and produced an anatomic picture of the false lumen. Three-dimensional imaging enhanced stent deployment by identifying the site for deployment, dimensions of the vessel lumen, and changes in morphology after stent insertion. There was good correlation in vessel dimensions measured by angiography, two-dimensional intravascular ultrasonography and longitudinal gray-scale reconstructions (r = 0.74 to 0.95; p = 0.34 to 0.001) but poor correlation with measurements from three-dimensional surface-rendered images (r = 0.13 to 0.48; p = 0.42 to 0.87). We conclude that three-dimensional intravascular ultrasound imaging is an innovative new method for identifying the type, extent, and spatial configuration of arterial disease, with promising diagnostic and therapeutic applications.

Aortic Dissection↗