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Biomedical subjects

P M Silverman

Publications and source records attributed to P M Silverman.

At least 19 recordsLinked to original sources

Interaction between the F plasmid TraA (F-pilin) and TraQ proteins.

Elaboration of conjugative (F) pili by F+ strains of Escherichia coli requires the activities of over a dozen F-encoded DNA transfer (Tra) proteins. The organization and functions of these proteins are largely unknown. Using the yeast two-hybrid assay, we have begun to analyse binary interactions among the Tra proteins required for F-pilus formation. We focus here on interactions involving F-pilin, the only known F-pilus subunit. Using a library of F tra DNA fragments that contained all the F genes required for F pilus formation in a yeast GAL4 activation domain vector (pACTII), we transformed yeast containing a plasmid (pAS1CYH2traA) encoding a GAL4 DNA-binding domain-F-pilin fusion. Doubly transformed cells were screened for GAL4-dependent gene expression. This screen repeatedly identified only a single Tra protein, TraQ, previously identified as a likely F-pilin chaperone. The F-pilin-TraQ interaction appeared to be specific, as no transcriptional activation was detected in yeast transformants containing pACTIItraQ plasmids and the Salmonella typhi pED208 traA gene cloned in pAS1CYH2. Two traQ segments isolated in the screen against F-pilin were tested for complementation of a traQ null allele in E. coli. One, lacking the first 11 (of 94) TraQ amino acids, restored DNA donor activity, donor-specific bacteriophage sensitivity and membrane F-pilin accumulation to wild-type levels. The second, lacking the first 21 amino acids, was much less effective in these assays. Both TraQ polypeptides accumulated in E. coli as transmembrane proteins. The longer, biologically active segment was fused to the GAL4 DNA-binding domain gene of pAS1CYH2 and used to screen the tra fragment library. The only positives from this screen identified traA segments. The fusion sites between the traA and GAL4 segments identified the hydrophobic, C-terminal domain IV of F-pilin as sufficient for the interaction. As TraQ is the only Tra protein required for the accumulation of inner membrane F-pilin, the interaction probably reflects a specific, chaperone-like function for TraQ in E. coli. Attempts to isolate an F-pilin-TraQ complex from E. coli were unsuccessful, suggesting that the interaction between the two is normally transient, as expected from previous studies of the kinetics of TraA membrane insertion and processing to F-pilin.

Amino Acid Sequence↗

Tuberculosis versus lymphomas in the abdominal lymph nodes: evaluation with contrast-enhanced CT.

OBJECTIVE: Tuberculosis in the abdominal lymph nodes may be difficult to distinguish from lymphomas. This study evaluated specific CT imaging criteria for differentiating these entities. MATERIALS AND METHODS: We retrospectively reviewed the anatomic distribution and CT enhancement patterns of disease in 69 patients, 26 (38%) with tuberculosis and 43 (62%) with untreated lymphomas involving abdominal lymph nodes. Of the patients with tuberculosis, five (19%) had disseminated disease and 21 (81%) had nondisseminated disease. Of the patients with lymphomas, 16 (37%) had Hodgkin's disease and 27 (63%) had non-Hodgkin's lymphoma. RESULTS: Disseminated and nondisseminated tuberculosis involved predominantly lesser omental, mesenteric, anterior pararenal, and upper paraaortic lymph nodes. Lower paraaortic lymph nodes were involved more often in Hodgkin's disease (15 patients [94%]), non-Hodgkin's lymphoma (24 patients [89%]), and disseminated tuberculosis (five patients [100%]) than in nondisseminated tuberculosis (one patient [5%]). Mesenteric lymph nodes were involved more often in disseminated tuberculosis (four patients [80%]) and nondisseminated tuberculosis (11 patients [52%]) than in Hodgkin's disease (one patient [6%]) (p < .01). Anatomic distribution was not different between disseminated tuberculosis and non-Hodgkin's lymphoma. Tuberculous lymphadenopathy commonly showed peripheral enhancement, frequently with a multilocular appearance, whereas lymphomatous adenopathy characteristically showed homogeneous attenuation (14 patients [87.5%] with Hodgkin's disease and 19 patients [70%] with non-Hodgkin's lymphoma [p < .01]). CONCLUSION: Our findings indicate that the anatomic distribution and specific enhancement patterns of lymphadenopathy seen on contrast-enhanced CT can be useful in differentiating between tuberculosis and untreated lymphomas of the abdominal lymph nodes.

Abdomen↗

Imaging of the liver with helical CT: a survey of scanning techniques.

OBJECTIVE: We conducted a survey of the members of the Society of Computed Body Tomography/Magnetic Resonance to assess current techniques in liver imaging using helical CT. MATERIALS AND METHODS: The survey, which was designed to update earlier surveys from 1987 and 1993, included a questionnaire distributed to 77 members of the Society of Computed Body Tomography/Magnetic Resonance. RESULTS: Forty-nine members responded, representing 28 institutions. In 1993, 19% (5/26) of institutions used helical scanners, compared with 82% (23/28) in 1996. The group of institutions with helical CT served as the focus of this survey. In 1993, 58% of institutions used 1-cm collimation: in 1996, 78% (18/23) used thinner, 7- to 8-mm collimation. In 1987, 41% used power injectors compared with 85% in 1993 and 100% in 1996. In 1996, monophasic injections were used by 96% (22/23) of institutions. In 1993, most institutions used a contrast material injection rate of 1.5-2.0 ml/sec; in 1996, most used a 2.5-3.0 ml/sec injection rate. In 1993, 96% of institutions used 125-150 ml of contrast material; in 1996, 57% (13/23) of institutions used 125-150 ml and 30% of institutions used less than 125 ml of contrast material. A delay time of 21-45 see was used by 83% of institutions in 1993, whereas in 1996, 83% (19/23) of institutions used a longer delay time of 50-80 sec. In 1996, 13% of institutions used an individual scan delay technology and all institutions performed multiphasic scanning of hypervascular lesions. CONCLUSION: The availability of helical CT has changed radiologists' approach to liver imaging. The greatest effects of which are a more widespread use of power injectors, longer delay times, thinner collimation, increased contrast material injection rates, decreased contrast material volumes, and multiphasic scanning.

Contrast Media↗

Peak enhancement of the liver in children using power injection and helical CT.

OBJECTIVE: Our objective was to determine the level and timing of peak hepatic enhancement in children using power injection of contrast media, helical CT, and computer-automated scan technology. SUBJECTS AND METHODS: Forty-nine abdominal CT studies were performed using computer-automated scan technology. Patients were divided into four groups on the basis of body weight and contrast dose (group 1A, < or = 20 kg and 2 ml/kg; group 1B, < or = 20 kg and 3 ml/kg; group 2, 21-40 kg and 2 ml/kg; group 3, > 40 kg and < or = 2 ml/kg). Contrast injection rates were based on body weight (groups 1A and 1B, 1 ml/sec; group 2, 1.5 ml/sec; and group 3, 2 ml/sec). The peak hepatic enhancement level in Hounsfield units and the time to reach peak enhancement were determined for each patient. RESULTS: The mean peak hepatic enhancement and time to peak enhancement after completion of contrast injection were group 1A, 45 H and 11 sec; group 1B, 62 H and 3 sec; group 2, 52 H and 12 sec; and group 3, 45 H and 10 sec. CONCLUSION: The level and timing of peak hepatic enhancement in pediatric patients can be obtained using computer-automated scan technology. These data may then be used to optimize hepatic enhancement when obtaining helical abdominal CT scans of pediatric patients.

Adolescent↗

Evaluation of an approach to choosing a left double-lumen tube size based on chest computed tomographic scan measurement of left mainstem bronchial diameter.

OBJECTIVE: Evaluation of an approach to choosing left double-lumen tube size based on chest computed tomographic (CT) scan measurement of left bronchial diameter. DESIGN: Prospective. SETTING: The operating rooms of a university hospital. PARTICIPANTS: Patients scheduled for elective thoracic surgery. INTERVENTIONS: Patients had their left bronchial diameter measured on the preoperative chest CT scan. Left double-lumen tube size for the individual patient was chosen from a protocol based on left bronchial diameter. MEASUREMENTS AND MAIN RESULTS: The double-lumen tube size was considered appropriate for the patient if some air leak was detected when the bronchial cuff was deflated and if airtight seal of the left bronchus was obtained with a bronchial cuff volume of 2 mL or less. In 17 of 20 patients, the double-lumen tube size fulfilled both criteria. In 3 women with left bronchi measuring 10 mm or less, the bronchus was sealed without any air in the bronchial cuff of size 35 Fr left double-lumen tubes. In 1 patient, who was excluded from the study, the double-lumen tube size was chosen based on measurement of the left bronchial diameter on chest radiograph because of motion artifact on the chest CT scan. CONCLUSIONS: Chest CT scan measurement of left bronchial diameter can successfully guide the choice of left double-lumen tube size for an individual patient. In individuals with a small left bronchus measuring less than 10.0 mm in diameter, currently available adult double-lumen tube sizes will tightly wedge in their bronchus.

Adult↗

Towards a structural biology of bacterial conjugation.

Horizontal DNA transfer among bacteria (conjugation) requires the formation of secure intercellular contacts before DNA transfer can occur. The formation of such contacts among Gram-negative bacteria is mediated by conjugative pili. Like other pili, conjugative pili are filaments extending from the surface of donor cells. They are composed, in so far as is known, entirely of conjugative pilin subunits. Here, we review the structure of F-pilin, the subunit of which F-pili are composed. We emphasize recent studies suggesting a specific domain organization for F-pilin and present a model of how these domains might be arranged in filament subunits.

Bacterial Outer Membrane Proteins↗

The greater omentum.

Understanding the embryologic development, anatomic relationships, and pathologic processes of the greater omentum is critical to its complete and proper evaluation. The broad spectrum of imaging findings presented in this essay may allow readers to appreciate features that aid accurate diagnosis of omental disease.

Adult↗

TNM staging of pancreatic carcinoma using helical CT.

OBJECTIVE: The purpose of this study was to determine the accuracy of helical CT scanning in predicting the stage of carcinoma of the exocrine pancreas using TNM staging guidelines and in predicting resectability of carcinoma of the exocrine pancreas. MATERIALS AND METHODS: Twenty-six patients with proven adenocarcinoma of the pancreas underwent uniphasic or biphasic helical CT scanning. Two observers unaware of the patient's surgical stage evaluated the CT examinations using the TNM system (with specific assessment and description of disease sites). In addition, the two observers rated confidence of nonresectability using a 5-point scale (ranging from 1, definitely resectable, to 5, definitely not resectable). Observer results and preoperative interpretations were compared with surgical findings. RESULTS: Nineteen of 26 patients had nonresectable disease. The combined observer scores showed correct determination of T stage in 77% of patients, of N stage in 58%, and of M stage in 79%. The overall accuracy in determining lack of resectability was 96% and 84% for the two observers. All errors in determining resectable versus nonresectable disease occurred when the observer was not maximally confident of his or her diagnosis. CONCLUSION: Helical CT is an effective screening technique for assessing T and M stages of pancreatic carcinoma. However, helical CT is poor at detecting regional lymph node involvement. In patients with equivocal T-stage findings (such as questionable venous involvement), other studies such as endoscopic sonography may be of value.

Adenocarcinoma↗

Effects of F-encoded components and F-pilin domains on the synthesis and membrane insertion of TraA'-'PhoA fusion proteins.

F-pilin, the 70-amino-acid F-pilus subunit, accumulates in the cell envelope of F+ strains in a process that requires interactions between its precursor (the traA gene product) and other host and F-encoded proteins. Here, we have used a set of phi(traA-phoA) genes to explore the effects of different TraA domains on the synthesis and membrane insertion of TraA-PhoA fusion proteins, particularly in relation to other F-encoded gene products. The 51-amino-acid TraA leader peptide fused directly to alkaline phosphatase was synthesized at comparable rates and incorporated rapidly and efficiently into the inner membrane in F' and F- cells. A second fusion gene encoded the TraA leader peptide and the first 51 amino acids of F-pilin itself fused to PhoA (TraA'-'PhoA-102 polypeptide). Alkaline phosphatase activities and patterns of pulse-labelled polypeptides indicated that TraA'-'PhoA-102 was synthesized at comparable rates in F' and F- cells, but in neither was the TraA'-'PhoA-102 polypeptide efficiently processed as a membrane protein. A third gene encoded the entire 121-amino-acid TraA polypeptide fused to PhoA (TraA-'PhoA-121 polypeptide). About 70% of the pulse-labelled TraA-'PhoA-121 polypeptide was rapidly processed in F' cells, where it accumulated in the cell envelope as active alkaline phosphatase, whereas in F- cells, < 5% of the pulse-labelled polypeptide was processed. Additionally, the apparent rate of TraA-'PhoA-121 polypeptide synthesis was threefold higher in F' cells. The traQ gene alone could not substitute for F in restoring TraA-'PhoA-121 (or wild-type F-pilin) accumulation.

Bacterial Outer Membrane Proteins↗

Effect of traY amber mutations on F-plasmid traY promoter activity in vivo.

We have examined the effect of the F plasmid TraY protein on tra gene expression in vivo. Expression was assayed as alkaline phosphatase activity in cells containing a traY phi(traA'-'phoA)hyb operon under traY promoter control. Amber mutations in traY significantly reduced alkaline phosphatase activity. Since nonsense polarity effects were minimal, if they occurred at all, these data provide the first direct evidence that TraY regulates tra gene expression.

Alkaline Phosphatase↗

Helical (spiral) CT of the upper airway with three-dimensional imaging: technique and clinical assessment.

OBJECTIVE: The purpose of this study was to evaluate the application of helical CT-generated three-dimensional images of the upper airway. MATERIALS AND METHODS: Thirty patients, 10 healthy and 20 with upper-airway disease, were studied with helical CT (5-mm collimation). Overlapping images at 2-mm intervals were retrospectively generated. In the group of healthy patients, two radiologists in independently compared overlapping with nonoverlapping images, ranked confidence in identifying small airway structures on a scale of 1-5, and tabulated the number of images demonstrating these structures. In the 20 patients with disease, three-dimensional (3D) surface models were rendered on an independent workstation and were reviewed by two radiologists and one otolaryngologist for image quality, appreciation of lesion morphology, and ability to judge lesion extent, using a similar scale. A phantom was used to optimize parameters for the 3D reconstructions. RESULTS: Viewing of the retrospectively generated overlapping images increased by 122% the number of images in which laryngeal and hypopharyngeal structures could be identified (p < .01). Image confidence scores for the radiologists averaged 3.3 for nonoverlapping and 4.0 for overlapping (p < .05). Radiologists and otolaryngologist rated the quality of the 3D images equally. The otolaryngologist's assessment of the value of the models for understanding the lesion morphology was 3.5 compared with the radiologists assessment of 2.5; and for judging the lesion extent, the otolaryngologist's assessment was 3.8 compared with 2.7 for the radiologist, a statistical significance of p < .01. CONCLUSION: Helical CT with the application of overlapping images and 3D reconstructions significantly assists the understanding of upper-airway disease.

Case-Control Studies↗

Low attenuation of acute traumatic hemoperitoneum on CT scans.

OBJECTIVE: This study was undertaken to determine the incidence of low attenuation values in intraperitoneal hemorrhage, which could be confused with ascites. MATERIALS AND METHODS: We retrospectively analyzed the CT scans of 42 consecutive patients with hepatic and splenic lacerations and intraperitoneal fluid after blunt abdominal trauma. Patients were excluded if they had prior peritoneal lavage, bladder or bowel injury, or low hematocrit values. Intraperitoneal fluid was categorized by the site of accumulation (perihepatic, perisplenic, Morison's pouch, paracolic gutters, or pelvis). The amount of fluid in each intraperitoneal location was categorized as small, moderate, or large. Attenuation values were obtained from each intraperitoneal site, and overall mean attenuation values were determined for each patient. We correlated the size of each fluid collection with the attenuation value. We also compared attenuation values at locations adjacent to the site of each injury with those at other intraperitoneal sites. We then evaluated technical factors that could have lowered attenuation values, including CT miscalibration, volume averaging, and beam-hardening artifacts. RESULTS: For the 42 patients, we measured 131 separate attenuation values. Attenuation values ranged from 0 to 80 H, with attenuation of 24% of sites (32/131) measuring less than 20 H. Only 16% of sites (21/131) had attenuation values greater than 45 H. Attenuation at the remaining 78 sites (60%) measured from 20 to 45 H. All intraperitoneal locations except the pelvis had mean attenuation values significantly lower then 40 H. Mean attenuation values (determined by averaging measurements from different intraperitoneal sites) were also calculated for each patient. Only 6 (14%) of 42 patients had mean attenuation values greater than 40 H, whereas 4 (10%) of 42 patients had mean attenuation values less than 20 H. The remaining 32 patients (76%) had mean attenuation values between 21 and 40 H. Patients with hepatic lacerations showed no significant difference (p = .3509) in attenuation between perihepatic fluid and the remainder of intraperitoneal fluid. However, in patients with splenic lacerations, perisplenic fluid had a significantly higher (p = .0013) attenuation value (43 H) than did fluid at other intraperitoneal locations. CONCLUSION: Low attenuation measurements for acute hemoperitoneum represented a common finding that was not attributable to technical factors or underlying anemia. Fluid with attenuation values less than 20 H in acute trauma should not be dismissed as ascitic fluid.

Abdominal Injuries↗