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Biomedical subjects

J E Reed

Publications and source records attributed to J E Reed.

At least 19 recordsLinked to original sources
PubMed2002

High-level expression of recombinant Fc chimeric proteins in suspension cultures of stably transfected J558L cells.

Recombinant Fc chimeric proteins are useful tools for studying protein function, including the analysis of molecular interactions by techniques such as expression cloning. Here we describe a method we have used to express the IgLON family proteins, CEPU1 and OBCAM, as recombinant Fc chimeric proteins in stably transfected mouse J558L myeloma cells. The use of this cell line provided the opportunity to maximize protein production, as it secretes antibodies in large quantities and can be grown to high density in small volumes of culture medium. Isolation of recombinant OBCAMFc from the adherent COS7 cell line suggested a minimum level of expression of 0.07 mg OBCAMFc/100 mL culture medium, while the J558L cell line expressed OBCAMFc at approximately 11.4 mg/100 mL culture medium. Purification of IgLON-Fc expressed by J558L cells was simpler than purification from COS7 cells because of the lower volume of culture medium generated. Furthermore, contamination of J558L expressed IgLONFc with bovine IgG from the culture medium was negligible. The method presented, which utilizes a commercially available small-scale bioreactor, provides the nonspecialist protein expression laboratory with the means to produce recombinant proteins quickly and easily in milligram quantities.

Animals↗
PubMed2001

A novel route to 2,3-pyrazol-1(5H)-ones via palladium-catalyzed carbonylation of 1,2-diaza-1,3-butadienes.

[reaction--see text] A novel Pd(0)-catalyzed carbonylation of both isolable 1,2-diaza-1,3-butadienes and those generated in situ by extrusion of SO(2) and CO(2) from heterocyclic precursors is described. The reaction proceeds at room temperature to 110 degrees C under 1-2 atm of CO to afford 2,3-pyrazol-1(5H)-ones in good to excellent yields. The effect of catalyst structure and stability on the carbonylation reaction is evaluated.

Alkenes↗
PubMed2001

Identification and characterization of CEPU-Se-A secreted isoform of the IgLON family protein, CEPU-1.

CEPU-1/Neurotrimin is a neuronal glycoprotein thought to play a role in axon guidance and cell-cell recognition. It is a member of the IgLON family, has three C2 domains, and is attached to the plasma membrane by a GPI-anchor. We report here the characterisation of an alternatively-spliced isoform of CEPU-1 that is secreted. This isoform, termed CEPU-Se, is coexpressed with CEPU-1 in retina, cerebellum, and DRG neurons. In the cerebellum CEPU-1/CEPU-Se is expressed predominantly on granule cells and in the molecular layer. Divalent but not monovalent CEPU-Se interacts with CEPU-1 and other IgLONs, suggesting that the ability of CEPU-Se to modify the activity of the IgLON family may require an additional cofactor. CEPU-Se does not support the outgrowth of DRG neurons or the extension of established growth cones; however, neurite outgrowth on laminin is unaffected by CEPU-Se. Our data suggest that CEPU-Se may act to modulate the ability of CEPU-1, LAMP, and OBCAM to influence neurite outgrowth.

Alternative Splicing↗
PubMed2001

Use of a new myocardial centroid for measurement of regional myocardial dysfunction by electron beam computed tomography: comparison with technetium-99m sestamibi infarct size quantification.

RATIONALE AND OBJECTIVES: The study compared the performance of conventional endocardial and epicardial centroid algorithms with the new "myocardial" centroid algorithm in patients with anterior myocardial infarction. "Floating" endocardial or epicardial centroid algorithms, commonly used in tomographic imaging methods to assess regional motion, may misrepresent left ventricular regional myocardial function in the presence of markedly asymmetric left ventricular contraction. METHODS: A new centroid algorithm based on regional myocardial mass distribution was tested in 29 patients with a first anterior myocardial infarction and was compared with conventional centroid algorithms. Direct comparisons in 60 equal sectors at one midventricular level per patient were performed between electron beam computed tomography and technetium-99m sestamibi single-photon emission computed tomography. The thresholds of regional myocardial function used to define infarction were varied for regional ejection fraction from 20% to 40% and for regional wall thickening from 0 to 4 mm. Regression and Bland-Altman analysis were used to compare infarct size by regional myocardial function with infarct size by sestamibi single-photon emission computed tomography. RESULTS: The new myocardial centroid showed the least shift toward infarcted myocardium from diastole to systole and had the highest amplitudes of the measurement curves for regional ejection fraction and regional wall thickening. The optimal regional myocardial function thresholds for each centroid algorithm for regional ejection fraction were endocardial, 30% (R = 0.62; mean difference to sestamibi, -0.5% +/- 22.1% tomographic infarct size points); epicardial, 30% (R = 0.79; mean difference, 2.2% +/- 13.1% tomographic infarct size points); and new myocardial, 25% (R = 0.88; mean difference, -0.6% +/- 9.5% tomographic infarct size points). The optimal thresholds for regional wall thickening were endocardial, 1 mm (R = 0.70; mean difference, -2.2% +/- 14.3% tomographic infarct size points); epicardial, 1 mm (R = 0.78; mean difference, -4.6% +/- 12.7% tomographic infarct size points); and new myocardial, 2 mm (R = 0.71; mean difference, 2.1% +/- 14.1% tomographic infarct size points). The best agreement (R = 0.88) between electron beam computed tomography infarct size and sestamibi single-photon emission computed tomography infarct size was achieved with regional ejection fraction and the new myocardial centroid algorithm. CONCLUSIONS: In asymmetrically contracting left ventricles, the new myocardial centroid algorithm is superior to conventional methods for tomographic analysis of regional myocardial function.

Algorithms↗
PubMed2001

Automated polyp detection at CT colonography: feasibility assessment in a human population.

PURPOSE: To test the feasibility of and improve a computer algorithm to automatically detect colonic polyps in real human computed tomographic (CT) colonographic data sets. MATERIALS AND METHODS: Twenty patients with known polyps underwent CT colonography in the supine position. CT colonographic data were processed by using a shape-based algorithm that depicts masses that protrude into the lumen. We studied nine shape criteria and three isosurface threshold settings. Results were compared with those of conventional colonoscopy performed the same day. RESULTS: There were 50 polyps (28 were > or =10 mm in size; 12, 5-9 mm; 10, <5 mm). The sensitivity with optimal settings for detecting polyps 10 mm or greater was 64% (18 of 28). Sensitivity improved to 71% (10 of 14) for polyps 10 mm or greater in well-distended colonic segments. Performance decreased for polyps less than 10 mm, poorly distended colonic segments, and other shape algorithms. There was a mean of six false-positive lesion sites per colon. These sites were reduced 39% to 3.5 per colon by sampling CT attenuation at the lesion site and discarding sites having attenuation less than a threshold. CONCLUSION: Automated detection of colonic polyps, especially clinically important large polyps, is feasible. Colonic distention is an important determinant of sensitivity. Further increases in sensitivity may be achieved by adding prone CT colonography.

Adult↗
PubMed2001

CT colonography without cathartic preparation: feasibility study.

PURPOSE: To evaluate methods for contrast material labeling of stool in the unprepared colon for computed tomographic (CT) colonography and to determine their sensitivity for polyp detection. MATERIALS AND METHODS: Fifty-six patients with suspected or known polyps were assigned to five groups. Two to seven doses of 225 mL of dilute contrast material were orally administered during 24 or 48 hours. Transverse CT images were assessed for effectiveness of stool labeling. Colonoscopy was performed in all patients and was the standard. Two radiologists blinded to prior imaging and colonoscopic results assessed polyp detection. RESULTS: For each group, average stool labeling scores and ranges were as follows: 24 hour two dose, 16% and 8%-21%; 24 hour five dose, 53% and 27%-66%; 48 hour four dose, 38% and 22%-48%; 48 hour six dose, 68% and 54%-77%; and 48 hour seven dose, 88% and 75%-98%. Sensitivity for the two radiologists for the identification of patients with polyps 1 cm or larger for each group was as follows: 24 hour two dose, 50% and 67%; 24 hour five dose, 100% and 100%; 48 hour four dose, 58% and 75%; 48 hour six dose, 56% and 67%; and 48 hour seven dose, 100% and 80%. CONCLUSION: Ingestion of contrast material adequately labels stool for lesion identification; a 48-hour lead time and multiple doses of contrast material are required. Sensitivity for polyp detection in patients with adequate stool labeling approaches the sensitivity for polyp detection in prepared colons.

Barium Sulfate↗
PubMed2000

Optimization of CT colonography technique: prospective trial in 180 patients.

PURPOSE: To assess the added benefits of prone positioning in addition to supine positioning and oral iodinated contrast medium for help in the detection of colonic polyps at computed tomographic (CT) colonography. MATERIALS AND METHODS: CT colonography was performed in prone and supine positions in 180 patients with polyps or risk factors for colonic neoplasia. Patients were randomly assigned to receive a standard bowel preparation or a standard preparation plus oral iodinated contrast medium. One radiologist interpreted supine images alone, and another analyzed supine and prone images. All patients subsequently underwent colonoscopy. RESULTS: At colonoscopy, 121 large (> or =1-cm-diameter) polyps and 142 smaller (0.5-0.9-cm) polyps were identified. Prone positioning resulted in increased sensitivity for identification of patients with large (> or =1-cm) polyps (increase from 70% to 85%, P: =.004) and of patients with polyps 0.5 cm or larger (increase from 75% to 88%, P: <.005), with no change in specificity. Use of oral contrast medium did not significantly improve polyp detection even in the subset of patients in whom colonic fluid attenuation was markedly increased. CONCLUSION: Acquisition and review of supine and prone CT colonographic images significantly improves the ability to identify patients with polyps 0.5 cm in diameter or larger. Administration of oral iodinated contrast medium does not significantly improve polyp detection.

Colon↗
PubMed2000

Measuring leg muscle and fat mass in humans: comparison of CT and dual-energy X-ray absorptiometry.

Dual-energy X-ray absorptiometry (DEXA) is reported to be inferior to computed tomography (CT) to measure changes in appendicular soft tissue composition. We compared CT- and DEXA-measured thigh muscle and fat mass to evaluate the random and systematic discrepancies between these two methods. Thigh skeletal muscle area (single-slice CT) was suboptimally (r(2) = 0.74, P < 0.0001) related to DEXA-measured thigh fat-free mass (FFM). In contrast, thigh muscle and adipose tissue volumes (multislice CT) were highly related to DEXA-measured thigh FFM and fat (both r(2) = 0.96, P < 0.0001). DEXA-measured leg fat was significantly less than multislice-CT-measured leg adipose tissue volume, whereas multislice-CT-measured leg muscle mass was less (P < 0.0001) than DEXA-measured leg FFM. The systematic discrepancies between the two approaches were consistent with the 10-15% nonfat components of adipose tissue. In conclusion, CT and DEXA measures of appendicular soft tissue are highly related. Systematic differences between DEXA and CT likely relate to the underlying principles of the techniques.

Absorptiometry, Photon↗
PubMed1999

An algorithm for noninvasive identification of angiographic three-vessel and/or left main coronary artery disease in symptomatic patients on the basis of cardiac risk and electron-beam computed tomographic calcium scores.

OBJECTIVES: We sought to model an algorithm for noninvasive identification of angiographically obstructive three-vessel and/or left main disease based on conventional cardiac risk assessment and site and extent of coronary calcium determined by electron-beam computed tomography (EBCT). BACKGROUND: Such an algorithm would greatly facilitate clinical triage in symptomatic patients with no previous diagnosis of coronary artery disease (CAD). METHODS: We examined 291 patients with suspected, but not previously diagnosed, CAD who underwent coronary angiography for clinical indications. Cardiac risk factors were determined as defined by the National Cholesterol Education Program. An EBCT scan was performed in all patients, and a coronary calcium score (Agatston method) was computed. Total per-patient calcium scores and separate scores for the major coronary arteries were generated. These scores were also analyzed for localization of coronary calcium in the more distal versus proximal tomographic sections. These parameters and the risk factors were considered for the model described in the following section. RESULTS: Sixty-eight patients (23%) had angiographic three-vessel and/or left main CAD. Multiple logistic regression analysis determined male sex, presence of diabetes and left anterior descending (LAD) and circumflex (LCx) coronary calcium scores, independent from more distal calcium localization, as independent predictors for identification of three-vessel and/or left main CAD. Based on this four variable model, a simple noninvasive index (NI) was constructed as the following: loge(LAD score) + log(e)(LCx score) + 2[if diabetic] + 3[if male]. Receiver operating characteristic curve analysis for this NI yielded an area under the curve of 0.88+/-0.03 (p < 0.0001) for separating patients with, versus without, angiographic three-vessel and/or left main CAD. Various NI cutpoints demonstrated sensitivities from 87-97% and specificities from 46-74%. The NI values >14 increased the probability of angiographic three-vessel and/or left main CAD from 23% (pretest) to 65-100% (posttest), and NI values <10 increased the probability of no three-vessel and/or left main CAD from 77% (pretest) to 95-100% (posttest). CONCLUSIONS: On the basis of a simple algorithm ("noninvasive index"), EBCT calcium scanning in conjunction with risk factor analysis can rule in or rule out angiographically severe disease, i.e., three-vessel and/or left main CAD, in symptomatic patients.

Algorithms↗
PubMed1999

Comparison of two different software systems for electron-beam CT-derived quantification of coronary calcification.

OBJECTIVE: The growing interest in coronary calcium quantification by electron-beam CT (EBCT) has led to the development of various software systems for the analysis of EBCT raw data, but it is unknown whether these software systems yield comparable results. METHODS: Two sets of EBCT scans were obtained in 73 asymptomatic patients less than 15 minutes apart. Both scans of each patient were analyzed using two different software systems, the Mayo Clinic software and the AccuImage Scoring System. The authors compared the calcium quantities yielded by the two different software systems, analyzed the interscan variability, and calculated the interobserver variability. Finally, they investigated the influence of the CT density factor inherent in the widely used Agatston score for the quantification of coronary calcium on reproducibility. RESULTS: The mean score determined by the Mayo Clinic software was 14% greater than that determined by the AccuImage system. The mean difference between the two systems was 14% +/- 25%, and the median difference was 3%. The relative mean and the median difference between the two scans of one patient were 15.3% and 6% determined by the AccuImage system and 17% and 6.5% determined by the Mayo Clinic software. The interobserver reliability calculated by the Mayo Clinic software was better than that of the AccuImage system. There was a trend for better reproducibility using calcium area rather than the Agatson score. CONCLUSIONS: Two different scoring systems do not necessarily yield the same result. Calcium quantities were systematically determined to be greater by one system than the other, and there were significant differences with regard to interobserver reliability. Hence, software should be tested with regard to reproducibility data, and the interpretation of calcium quantities should acknowledge which type of software was used.

Adult↗
PubMed1998

Pituitary desensitization to gonadotropin-releasing hormone increases abdominal adiposity in hyperandrogenic anovulatory women.

OBJECTIVE: To determine whether hyperandrogenism in anovulatory women affects body fat distribution. DESIGN: Prospective nonrandomized study. SETTING: An academic research environment. PATIENT(S): Ten hyperandrogenic anovulatory patients and 10 healthy women matched by body mass index. INTERVENTION(S): Regional body fat analysis was performed before and after 3 months of GnRH analogue (GnRH-a) therapy. MAIN OUTCOME MEASURE(S): Body fat distribution was measured by waist-to-hip circumference ratio, single-slice computed tomography imaging (L2-3 interspace), and total body dual-energy x-ray absorptiometry. RESULT(S): Weight, body mass index, waist-to-hip circumference ratio, total body and leg fat mass, and subcutaneous adipose area were unaffected by the presence of hyperandrogenism or the use of GnRH-a therapy. Basal abdominal fat mass, abdomen-to-leg fat mass ratio, visceral adipose area, and total visceral adipose volume were comparable in both study groups. The abdominal fat mass increased in both groups during GnRH-a therapy, whereas the abdomen-to-leg fat mass ratio rose significantly only in the hyperandrogenic patients. During GnRH-a therapy, the hyperandrogenic patients demonstrated a significant increase in visceral adipose area compared with the healthy women so that total visceral adipose volume increased significantly in the former but not the latter. CONCLUSION(S): Three months of GnRH-a administration preferentially increased abdominal fat, as measured by single-slice computed tomography imaging and total body dual-energy x-ray absorptiometry, in hyperandrogenic anovulatory women.

Abdomen↗
PubMed1998

Measurement of myocardial infarct size by electron beam computed tomography: a comparison with 99mTc sestamibi.

RATIONALE AND OBJECTIVES: The authors sought to determine, using a variety of regional left ventricular ejection fraction (EF) and wall thickening (WTh) criteria, the applicability to measure left ventricular (LV) infarct size using electron-beam CT (EBCT) in patients as compared with technetium 99m (99mTc) sestamibi scanning as reference standard. METHODS: Twelve patients (age 57 +/- 11 years) underwent 99mTc sestamibi scanning and EBCT at hospital discharge after an acute index anterior myocardial infarction. Left ventricular infarct size was defined using standard 99mTc sestamibi scanning. Regional EF and WTh were analyzed on each EBCT scan with use of a floating epicardial centroid method. In five contiguous LV tomograms, the amount of infarcted myocardium was estimated using the following EF and WTh criteria: EF < or = 35%, 30%, 25%, 20%, and WTh < or = 2 mm, 1 mm, and 0 mm. RESULTS: Infarct size measured with 99mTc sestamibi was 33.3% (+/- 18.3%) (mean +/- SD, range 6%-54%) of the LV. Using an EF < or = 35% or absolute WTh < or = 2 mm as criteria for infarcted myocardium, EBCT yielded 28% (+/- 17%) and 27% (+/- 16%), respectively (P = NS, paired Student's t test, versus 99mTc sestamibi). Although, with use of the other criteria, EBCT tended to underestimate infarct size compared with 99mTc sestamibi, a close correlation across the entire range of infarct size determinations (range, 0.72-0.82) regardless of the underlying criteria suggested an internal consistency of the data. CONCLUSIONS: Quantitative analysis of regional myocardial function by EBCT allows an estimate of anterior infarct size when compared with 99mTc sestamibi. This suggests that in addition to previously established applications after acute myocardial infarction such as examination of cardiac volumes and mass, EBCT also may provide for infarct size determination.

Adult↗
PubMed1997

Automatic segmentation, tissue characterization, and rapid diagnosis enhancements to the computed tomographic colonography analysis workstation.

An image processing system developed to support examination of computed tomographic colonoscopy (CTC) was developed in 1995. The clinical viability of CTC is enhanced by the solution of several technical problems. These problems include the limited detectability of sessile polyps and difficulties in discrimination between polypoid masses and retained stool. CTC is also made more feasible by simplifying the required colon preparation and reducing the time required to analyze scan results. Each of these challenges have been addressed by enhancements to the CTC analysis workstation software. Endoluminal volume rendering has been enhanced by the addition of automatic segmentation to facilitate analysis of colon segments, which contain tagged liquid stool. By automating this function, the system is able to process scans that are acquired following a wide variety of colon preparation protocols. Similar approaches have been used to identify retained stool. Automatic tissue characterization has also been incorporated into the volume rendering routines to help identify and diagnose polypoid masses. These enhancements have improved the quality of CTC interpretation, while reducing the time required to perform the analysis. This time reduction was necessary to reduce the cost of CTC enough to make it viable for asymptotic population screening. To date, over 150 patient examinations have been performed using this new technique. A recent blinded, prospective study reporting the results from two independent observers has been presented. The technique is feasible, reliable, and has been implemented clinically with results reported within 1 hour of the examination.

Colon↗
PubMed1997

Computed tomographic colonography (Virtual colonoscopy): a new method for detecting colorectal neoplasms.

Computed tomographic (CT) colonography is an exciting new technique that uses volumetric CT data combined with advanced imaging software to create two-dimensional and three-dimensional images of the colon. The technique uses both three-dimensional images that simulate the endoluminal perspective of the colonoscope, as well as axial and reformatted two-dimensional images. The two-dimensional and three-dimensional images are complementary, and in combination offer the most robust performance for the detection of colorectal polyps. Currently, CT colonographic examinations are performed in the fully cleansed and air-inflated colon using a slice thickness of 5 mm, a reconstruction interval of 3 mm, a pitch of 1.3, and 70 mA. In a blinded, prospective study of 70 patients (half with a known lesion, and half from a surveillance population with a low disease prevalence) the sensitivity for the detection of polyps of 1 cm or more is 75%, and the specificity is 90%. The most commonly encountered problems include retained colonic fluid and stool, suboptimally distended colonic segments, and long interpretation times. Many of these problems can be solved using both supine and prone imaging. It is expected that the performance of this examination will improve, and that a new era of colorectal screening will begin.

Colonoscopy↗