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D G Heath

Publications and source records attributed to D G Heath.

34 records · Page 2Linked to original sources
PubMed1995

Phase variation of Enterococcus faecalis pAD1 conjugation functions relates to changes in iteron sequence region.

pAD1 (60 kb) is a conjugative, hemolysin/bacteriocin plasmid in Enterococcus faecalis. It confers a mating response to the peptide sex pheromone cAD1 produced by recipient (plasmid-free) cells, leading to highly efficient plasmid transfer in broth matings. Control of the physiological response to cAD1 can been overridden by a reversible phase variation event at frequencies on the order of 10(-4) to 10(-3) per cell per generation (L. T. Pontius and D. B. Clewell, Plasmid 26:172-185, 1991). The variant forms are designated Dryc and Dry+, which reflects the colony morphologies of cells whose conjugation functions are switched on and off, respectively. Here we show that Dryc variants exhibit a structural change in a region between repA and repB that contains two clusters of 8-bp iterons. The change involved a 31- or 32-bp increase in size of this region. In three or four independent variants examined, one of the iteron clusters increased in size from 13 to 17 iterons. When iteron DNA was placed on a multicopy plasmid and introduced into a wild-type pAD1 derivative, the Dryc phenotype was generated. Since traA, a key negative regulator of conjugation, bears several centrally located iteron-like sequences with the same orientation, we speculate that the protein(s) that normally binds iterons (possibly RepA and/or RepB) blocks traA transcription in Dryc variants.

Bacterial Proteins↗
PubMed1995

Three-dimensional spiral CT during arterial portography: comparison of three rendering techniques.

The three most common techniques for three-dimensional reconstruction are surface rendering, maximum-intensity projection (MIP), and volume rendering. Surface-rendering algorithms model objects as collections of geometric primitives that are displayed with surface shading. The MIP algorithm renders an image by selecting the voxel with the maximum intensity signal along a line extended from the viewer's eye through the data volume. Volume-rendering algorithms sum the weighted contributions of all voxels along the line. Each technique has advantages and shortcomings that must be considered during selection of one for a specific clinical problem and during interpretation of the resulting images. With surface rendering, sharp-edged, clear three-dimensional reconstruction can be completed on modest computer systems; however, overlapping structures cannot be visualized and artifacts are a problem. MIP is computationally a fast technique, but it does not allow depiction of overlapping structures, and its images are three-dimensionally ambiguous unless depth cues are provided. Both surface rendering and MIP use less than 10% of the image data. In contrast, volume rendering uses nearly all of the data, allows demonstration of overlapping structures, and engenders few artifacts, but it requires substantially more computer power than the other techniques.

Algorithms↗
PubMed1992

Identification of two type IIa IgG-binding proteins expressed by a single group A streptococcus.

Functional heterogeneity associated with Ig-binding proteins expressed by group A streptococci is well documented. In this study we have demonstrated that treatment of group A streptococcal isolate 64/14 with CNBr resulted in the solubilization of two different sized proteins that displayed identical functional reactivity with human IgG1, IgG2, and IgG4 (characteristics of a type IIa binding protein). Monospecific polyclonal antibodies to each form of type IIa molecule were prepared and no antigenic cross-reactivity between the two m.w. forms of type IIa binding protein could be detected. The smaller m.w. protein was shown to be identical or closely related to the recombinant type IIa protein cloned from strain CS110. These studies provide further evidence for the heterogeneity of type II Ig-binding proteins expressed by pathogenic group A streptococci.

Bacterial Proteins↗
PubMed1992

Architecture of the vir regulons of group A streptococci parallels opacity factor phenotype and M protein class.

Group A streptococci have traditionally been categorized into two broad groups based on the presence or absence of serum opacity factor (OF). Recent studies show that these two groups vary in a number of properties in addition to the OF phenotype, including sequence variations in the constant region of the antiphagocytic M protein genes, the presence or absence of immunoglobulin G Fc receptor proteins, and the presence or absence of multiple M protein-like genes situated in a tandem array. The M protein genes (emm) in OF- streptococcal strains are known to be part of a regulon of virulence-related genes controlled by the trans-acting positive regulatory gene, virR, situated just upstream of emm. In OF+ strains, however, the region adjacent to virR is occupied by an M protein-related, type IIa immunoglobulin G Fc receptor gene (fcrA), and the relative position of emm has not been determined. To further define the vir regulon in OF+ streptococci, we used the polymerase chain reaction to show that fcrA49 is situated immediately upstream of emm49 in the OF+ type 49 strain CS101. This result shows for the first time the separate identity and genetic linkage of these two genes in the vir regulon of an OF+ group A streptococcal strain and confirms our previous hypothesis that emm49 exists as the central gene in a trio of emm-like genes. Additionally, using DNA hybridizations, we found considerable sequence divergence between OF- and OF+ group A streptococci in virR and in the noncoding sequences between virR and the emm or fcrA expression site. We found, however, a high degree of sequence conservation in this region within each of the two groups of strains.

Amino Acid Sequence↗
PubMed1992

Degenerate oligonucleotide primers for enzymatic amplification of recA sequences from gram-positive bacteria and mycoplasmas.

RecA protein in gram-negative bacteria, especially in Escherichia coli, has been extensively studied, but little is known about this key enzyme in other procaryotes. Described here are degenerate oligonucleotide primers that have been used to amplify by the polymerase chain reaction (PCR) recA sequences from several gram-positive bacteria and mycoplasmas. The DNA sequences of recA PCR products from Streptococcus pyogenes, Streptococcus mutans, Enterococcus faecalis, and Mycoplasma pulmonis were determined and compared. These data indicate that the M. pulmonis recA gene has diverged significantly from recA genes of other eubacteria. It should be possible to use cloned recA PCR products to construct recA mutants, thereby providing the means of elucidating homologous genetic recombination and DNA repair activities in these organisms.

Amino Acid Sequence↗
PubMed1990

Isolated DNA repeat region from fcrA76, the Fc-binding protein gene from an M-type 76 strain of group A streptococci, encodes a protein with Fc-binding activity.

The DNA repeat region of fcrA76, the gene encoding a group A streptococcal Fc-binding protein, was subcloned in-frame into an Escherichia coli plasmid expression vector. The expressed protein product displayed the same Fc-binding properties as the full-length Fc-binding protein expressed from fcrA76. The affinity-purified, full-length Fc-binding protein was found to compete with staphylococcal protein A or streptococcal protein G for binding to beads coated with human IgG. These results are consistent with earlier studies suggesting that the binding sites on human IgG for protein A, protein G and the type II Fc-binding protein from group A streptococci are located at the interface of the CH2 and CH3 domains of the Fc region.

Amino Acid Sequence↗
PubMed1989

Fc-receptor and M-protein genes of group A streptococci are products of gene duplication.

The partial nucleotide sequence for an Fc-receptor gene from an M-type 76 group A streptococcus was determined. DNA sequence analysis revealed considerable sequence similarity between the Fc-receptor and M-protein genes in their proposed promoter regions, signal sequences, and 3' termini. Additional analysis indicated that the deduced Fc-receptor protein contains a proline-rich region and membrane anchor region highly similar to that of M protein. In view of these results, we postulated that Fc-receptor and M-protein genes of group A streptococci are the products of gene duplication from a common ancestral gene. It is proposed that DNA sequence similarity between these two genes may allow for extragenic homologous recombination as a means of generating antigenic diversity in these two surface proteins.

Amino Acid Sequence↗
PubMed1987

Cloning and expression of the gene for an immunoglobulin G Fc receptor protein from a group A streptococcus.

DNA containing the 5' end of the M-12 structural gene was used as a probe in colony hybridizations in an attempt to clone the M-76 gene from an M-type 76 group A streptococcal strain. A single positive colony was detected, and Southern hybridization analysis of plasmid DNA isolated from this colony indicated that the insert DNA had homology to the 5' end of the M-12 structural gene. Subclones were constructed to define the limits of the M-76 gene, and sonicates of these subclones were reacted with M-76-specific antiserum in immunodiffusion. A sonicate of one subclone, JM83(pDH56), reacted strongly with the M-76-specific antiserum but also reacted with preimmune rabbit serum. Protein expressed from this subclone bound immunoglobulin from horse and pig, as well as human myeloma immunoglobulin G (IgG) representing all four subclasses and purified human IgG Fc fragments. This indicated that JM83(pDH56) expressed a protein with characteristics previously attributed to the IgG Fc receptor protein from group A streptococci. Western blot analysis indicated that the cloned IgG Fc receptor protein had a molecular weight of approximately 29,000. Binding studies showed that the Fc receptor gene is expressed by the M-type 76 strain from which it was cloned and by an M- variant.

Cloning, Molecular↗
PubMedDate not supplied

Virtual angioscopy using spiral CT and real-time interactive volume-rendering techniques.

Our purpose was to describe a technique for visualizing the inner contours of the vasculature using contrast enhanced spiral CT and volume rendering techniques. Because the technique is similar to using a camera to look inside vessels, we call this technique "virtual angioscopy." Preliminary results suggest virtual angioscopy using volumetric 3D rendering techniques as a potentially useful technique for the noninvasive evaluation of vascular pathology.

Adult↗
PubMedDate not supplied

Dual-phase spiral CT angiography with volumetric 3D rendering for preoperative liver transplant evaluation: preliminary observations.

PURPOSE: The goal of our study was to determine whether dual-phase spiral CT angiography with 3D volume rendering could be used for preoperative evaluation and patient selection for orthotopic liver transplantation candidates. METHOD: Fifty consecutive potential candidates for liver transplantation were evaluated with dual-phase spiral CT with 3D volume rendering. Intravenous contrast medium was administered as bolus peripheral injection at 3 ml/s. The protocol consisted of a contrast-enhanced dual-phase spiral CT (arterial phase acquisition at 30 s after initiation of contrast medium injection followed by portal venous phase beginning at 60 s) with scan parameters of 0.75 s gantry rotation speed, 3 mm collimation, 5 to 6 mm/s table speed, and reconstruction at 1 mm intervals for arterial-phase images and 3 mm collimation for portal venous-phase studies (Siemens Plus 4 scanner; Siemens Medical Systems, Iselin, NJ, U.S.A.). All scan information was sent to a free-standing workstation (Silicon Graphics Onyx or Infinite Reality, Mountain View, CA, U.S.A.) for interactive real-time 3D volume rendering using a customized version of the Volren volume renderer (Silicon Graphics; Advanced Imaging Laboratory, Johns Hopkins Medical Institutions, Baltimore, MD, U.S.A.). The arterial phase was used to create vascular maps of the celiac axis including the origin(s) of the hepatic artery and origin of the superior mesenteric artery. The portal phase was used to define portal venous patency as well as the hepatic venous anatomy. All images were analyzed for vascular patency, shunting, or collateralization as well as the status of the underlying liver (i.e., liver size, cirrhosis, tumor, etc.). RESULTS: All 50 studies were successfully completed without complication. The 3D CT angiograms defined key arterial and venous structures including origin(s) of the hepatic artery, portal vein and/or superior mesenteric vein thrombosis, cavernous transformation of the portal vein, and/or other collateral vasculature. Ten patients (20%) demonstrated anomalous anatomy at the origin(s) of the hepatic artery. Portal vein thrombosis with cavernous transformation of the portal vein was shown in six patients, and there were three cases of partial venous thrombosis. Underlying liver tumors as well as parenchymal liver disease were well defined. Hepatic masses were found in five patients. Masses were pathologically proven as hepatocellular carcinoma (n = 1), giant cavernous hemangioma (n = 1), hepatic adenoma (n = 1), and focal nodular hyperplasia (n = 2). CONCLUSION: Preliminary results suggest that dual-phase spiral CT with CT angiography can provide a comprehensive preoperative liver transplant evaluation, supplying the necessary information for patient selection and surgical planning. As a single, minimally invasive examination, this should significantly impact patient care by minimizing procedures and avoiding potential complications.

Adolescent↗
PubMedDate not supplied

Interactive three-dimensional volume rendering of spiral CT data: current applications in the thorax.

Spiral computed tomographic (CT) data sets coupled with a real-time volume-rendering technique allow creation of accurate three-dimensional (3D) images that can be used for a wide range of clinical applications. The image contrast of and relative pixel attenuations in the final image can be interactively modified by the user in real time by manipulating trapezoidal transfer functions. Although 3D images are not required for diagnosis, they aid both radiologists and referring clinicians by demonstrating anatomic relationships and the extent of disease, particularly for vessels oriented in the z axis. Three-dimensional imaging of the vasculature and airway structures has many advantages, including the potential to obviate invasive procedures such as angiography and bronchoscopy. Clinical applications of volume rendering of spiral CT data include cardiovascular imaging (aorta, pulmonary vasculature, and venous abnormalities), staging of thoracic neoplasms (mediastinal and pulmonary masses), tracheobronchial imaging, and imaging of chest wall disease.

Adult↗
PubMedDate not supplied

Three-dimensional volume rendering of spiral CT data: theory and method.

Three-dimensional (3D) medical images of computed tomographic (CT) data sets can be generated with a variety of computer algorithms. The three most commonly used techniques are shaded surface display, maximum intensity projection, and, more recently, 3D volume rendering. Implementation of 3D volume rendering involves volume data management, which relates to operations including acquisition, resampling, and editing of the data set; rendering parameters including window width and level, opacity, brightness, and percentage classification; and image display, which comprises techniques such as "fly-through" and "fly-around," multiple-view display, obscured structure and shading depth cues, and kinetic and stereo depth cues. An understanding of both the theory and method of 3D volume rendering is essential for accurate evaluation of the resulting images. Three-dimensional volume rendering is useful in a wide variety of applications but is just now being incorporated into commercially available software packages for medical imaging. Although further research is needed to determine the efficacy of 3D volume rendering in clinical applications, with wider availability and improved cost-to-performance ratios in computing, 3D volume rendering is likely to enjoy widespread acceptance in the medical community.

Algorithms↗