Biomedical subjects

B J Wagner

Publications and source records attributed to B J Wagner.

At least 19 recordsLinked to original sources

PubMed1999

Identification of a novel gene product preferentially expressed in rat lens epithelial cells.

PURPOSE: We searched for an mRNA that is differentially expressed in rat lens epithelial and fiber cells to use as a marker for differentiation. METHODS: An mRNA differential display method was used to identify genes with altered expression during differentiation of lens epithelial cells. RESULTS: One gene (partial sequence) was identified and confirmed to be expressed preferentially in undifferentiated rat lens epithelial cells. Among non-lens tissues, a trace amount was detected in cornea, but not in retina or eleven non-ocular tissues tested. This cloned partial gene sequence (322 base pairs) is at least 93% identical to overlapping rat and mouse EST sequences, and extends the sequence of both ESTs at the 5' end. CONCLUSIONS: A clone has been isolated that is preferentially expressed in lens epithelial cells. Expression of this gene was down-regulated when rat pup lens epithelial cell explants were induced to differentiate. The unusual pattern of expression suggests a novel function for this gene.

Animals↗

PubMed1999

Large degenerated adrenal adenomas: radiologic-pathologic correlation.

PURPOSE: To correlate the radiologic and pathologic findings and differential diagnosis of large, degenerated adrenal adenomas. MATERIALS AND METHODS: The authors reviewed the radiologic and pathologic characteristics of 30 large adenomas with cystic regions or areas of heterogeneity that were either intrinsic or demonstrated at contrast material-enhanced computed tomography (CT) or magnetic resonance (MR) imaging. Images of 24 adrenocortical carcinomas were also reviewed to determine whether differentiating characteristics existed. RESULTS: Most of the adrenocortical adenomas were in asymptomatic women. Ten adenomas contained calcification. Pathologic examination revealed good correlation between heterogeneity and liquefied regions. Histologic examination confirmed regions of adenomatous tissue with areas of hemorrhage, amorphous degenerated material, calcification, and fibrosis. Some tumors contained myelolipomatous foci. Although some clinical and imaging findings differed between the groups, no features could be found that enabled the radiologic differentiation of adenomas from carcinomas. CONCLUSION: A subgroup of adrenal adenomas are larger, more heterogeneous, and more frequently calcified than those with the usual imaging findings. Central necrosis, hemorrhage, or both are responsible for many of the imaging features. Differentiation of these lesions from other large adrenal masses, including adrenal carcinoma, cannot be made by means of imaging alone; resection is required for the definitive diagnosis.

Adrenal Cortex↗

PubMed1999

Aggressive angiomyxoma: findings on CT and MR imaging.

OBJECTIVE: Aggressive angiomyxoma is a benign tumor affecting the pelvis and perineum, predominantly in women. Because of its variable presentation as a soft mass in the vulva, perianal region, buttock, or pelvis, the tumor is often clinically misdiagnosed and initial surgery is usually unsuccessful in extirpating it. This study describes the imaging features of these tumors. CONCLUSION: Aggressive angiomyxomas display unusual growth patterns of translevator extension with growth around perineal structures. Both CT and MR imaging show the transdiaphragmatic extent of these tumors. High signal intensity on T2-weighted MR images may reflect the myxomatous stroma of these tumors.

Adult↗

PubMed1998

Structure and genomic organization of the human AUF1 gene: alternative pre-mRNA splicing generates four protein isoforms.

The steady-state levels of many mRNAs are determined in part by their turnover rates. Turnover rates, in turn, are usually controlled by proteins that bind cis-acting sequence elements in mRNAs. One class of cis-acting instability determinants is composed of A + U-rich elements present in the 3'-UTRs of many labile mRNAs. Many A + U-rich elements are bound by the AUF1 family of RNA-binding proteins, which may target these mRNAs for rapid decay. cDNA cloning and immunoblot analyses suggest that the AUF1 family consists of at least four isoforms. Previous genomic cloning combined with FISH and Southern analyses of a panel of monochromosomal mouse/human or hamster/human somatic cell hybrids localized two AUF1 loci to human 4q21.1-q21.2 and Xq12 (B. Wagner et al., 1996, Genomics 34: 219-222). In the present study AUF1 gene organization was examined. The results suggest that the four known AUF1 isoforms are generated by alternative pre-mRNA splicing of a transcript encoded by the chromosome 4 locus. Functionally, this creates isoforms with different RNA-binding affinities and specificities. Thus, alternative pre-mRNA splicing may serve to create functional versatility within the AUF1 family of proteins.

Alternative Splicing↗

PubMed1998

Gene expression of the proteasome in rat lens development.

To study the involvement of the proteasome in ocular lens cell proliferation and differentiation, a partial cDNA encoding rat S7, a subunit of the ATPase complex that regulates the 20S proteasome (multicatalytic proteinase complex), and RC3, a subunit of the 20S proteasome moiety, were cloned and used to compare relative levels of S7 and RC3 mRNAs. mRNA was measured, using a competitive RT-PCR assay, in isolated lens cells or explant cultures induced to differentiate or proliferate. During differentiation, S7 mRNA levels increased (1.7 fold) and RC3 mRNA levels remained the same compared to mRNA in quiescent cells. During proliferation, RC3 mRNA levels were elevated (2 fold) and S7 mRNA levels remained the same. This demonstrated that representative proteasome and ATPase complex mRNA levels are regulated differentially during differentiation and proliferation. The maintenance of proteasome subunit mRNA and increase in ATPase complex subunit mRNA observed in differentiating lens cells is in contrast to the patterns of expression that have been reported for other differentiating cells, which down-regulate the 20S and/or 26S proteasome. This suggests that the role of the proteasome in cell development is cell specific.

Acetylcysteine↗

PubMed1998

Myelolipoma: CT and pathologic features.

PURPOSE: To correlate clinical and pathologic patterns of myelolipoma with the computed tomographic (CT) appearance. MATERIALS AND METHODS: All cases of myelolipoma accessioned by the Armed Forces Institute of Pathology from 1981 through 1997 were reviewed. CT images were scored for the location and size of each myelolipoma and the presence of calcification, hemorrhage, fat, and pseudocapsule. Pathologic findings for the pattern of fat and bone marrow elements were correlated with CT findings. RESULTS: In 74 patients, 86 myelolipomas were found, of which 72 were in an adrenal gland (eight were bilateral), and 14 extra-adrenal masses were found in 10 patients. Four clinicopathologic patterns emerged: (a) isolated adrenal myelolipoma in 37 patients (fat evident at CT, no other disorders present); (b) myelolipoma with hemorrhage in nine patients (imaging features similar to those of isolated adrenal myelolipomas but larger [mean diameter, 14.2 vs 9.9 cm; P = .01]; (c) extra-adrenal myelolipoma in 10 patients (imaging findings similar to those of adrenal myelolipomas, found most often in the retroperitoneum); and (d) myelolipomatous foci within other adrenal pathologic conditions in 18 patients (smaller, lower fat content, more heavily calcified). CONCLUSION: Myelolipomas are adrenal or extra-adrenal masses, with hemorrhage more common in larger lesions (diameter, > 10 cm). The CT appearance of myelolipomatous foci, which can be found within other pathologic adrenal conditions, is different from that of isolated adrenal myelolipomas.

Adipose Tissue↗

PubMed1997

Structural determinants in AUF1 required for high affinity binding to A + U-rich elements.

AUF1 is an RNA-binding protein that contains two nonidentical RNA recognition motifs (RRMs). AUF1 binds to A + U-rich elements (AREs) with high affinity. The binding of AUF1 to AREs is believed to serve as a signal to an mRNA-processing pathway that degrades mRNAs encoding many cytokines, oncoproteins, and G protein-coupled receptors. Because the ARE binding activity of AUF1 appears central to the regulation of many important genes, we analyzed the domains of the protein that are important for this activity. Examination of the RNA binding affinity of various AUF1 mutants suggests that both RRMs may be required for binding to the human c-fos ARE. However, the two RRMs together are not sufficient. Highest affinity binding of AUF1 to an ARE requires an alanine-rich region of the N terminus and a short glutamine-rich region in the C terminus. In addition, the N terminus is required for dimerization of AUF1. However, AUF1 binds an ARE as a hexameric protein. Thus, protein-protein interactions are important for high affinity ARE binding activity of AUF1.

Adenine↗

PubMed1997

Radiologic assessment of renal masses: implications for patient care.

The relationships between the gross pathologic features of neoplastic and nonneoplastic renal masses and their radiologic analogues, described above, establish specific guidelines for the influence of radiologic studies on clinical management. A tumor that contains fat, as determined with CT or MR imaging, can be confidently diagnosed as an angiomyolipoma without further diagnostic intervention. The size of the lesion should be used to influence clinical decisions related to the fact that angiomyolipomas larger than 4 cm in diameter are more apt to hemorrhage than those smaller than that size. High confidence can also be assigned to those renal masses that exhibit the radiologic analogues for hemangioma with use of imaging modalities that document their vascular nature. These findings should be sufficient for therapeutic decisions directed toward embolization or surgical excision when clinically warranted. If a mass can be characterized as a simple cyst by satisfying all of the required CT or sonographic criteria, no further diagnostic interventions are required. This includes the radiologic findings of thin rim of peripheral calcification and thin septa with or without calcification. An equally high level of confidence is associated with the broad range of CT, sonographic, or MR imaging findings that indicate malignant tumor. These militate for radical surgery. However, the same findings are also encountered in hemorrhagic and infected renal cyst, abscess, benign neoplasms, and inflammatory mass. Therefore, surgical excision, the nature of which will vary according to individual circumstances, is usually required to establish these diagnoses. Exceptions to the need for a tissue diagnosis might be considered in the patient in whom a renal mass is detected in the clinical setting of infection and in the patient with either a small asymptomatic renal mass or a small hyperattenuating mass that meets the other criteria of a simple cyst. Here again, individual circumstances may lead to such alternatives as aspiration of the mass for culture, interval follow-up to seek evidence of growth, or dismissal.

Abscess↗

PubMed1997

Rhabdoid tumor of the kidney in children: a comparative study of 21 cases.

OBJECTIVE: Rhabdoid tumor of the kidney (RTK) is unique among childhood renal neoplasms in its frequent association with primary or metastatic CNS lesions. Previous reports suggest that RTK has characteristic CT features. It has been proposed that if CT could accurately predict the correct diagnosis of RTK preoperatively, then imaging protocols might be modified during the examination to include imaging of the brain. We wished to determine if RTK could be reliably distinguished from other renal neoplasms of early childhood. MATERIALS AND METHODS: We retrospectively reviewed clinical, radiologic, and pathologic records of 21 patients with RTK. The study group included 13 males and eight females who were newborn to 36 months old (mean, 11 months). The study group was compared with 153 patients who were 3 years old or younger and who had solid renal masses. From the comparison group a subset of 54 patients 1 year old and younger was also selected for comparison with 13 (62%) of the 21 patients in the study group who were 1 year old or younger. Both comparison groups consisted of patients whose case material was consecutively added to the radiologic pathology archives at our institution. Diagnoses of the group of 153 patients were Wilms' tumor (n = 93), mesoblastic nephroma (n = 44), clear cell sarcoma of the kidney (n = 12), renal cell carcinoma (n = 3), and undifferentiated sarcoma (n = 1). RESULTS: A prominent eccentric crescent with the attenuation of fluid, representing sub-capsular renal hemorrhage or peripheral tumor necrosis adjacent to tumor lobules, was revealed on CT scans in 15 (71%) of the 21 patients with RTK and in seven (54%) of the 13 patients with RTK who were 1 year old or younger. Nineteen (12%) of 153 patients in the larger comparison group, representing patients with all tumor types, had CT features identical to those of the RTK group, including six (4%) patients with pathologic confirmation of sub-capsular renal hematomas. Six (11%) of the 54 comparison patients 1 year old and younger had CT features identical to those of the RTK group, and all proved to have mesoblastic nephroma. Associated CNS lesions were seen on CT or MR imaging in 11 (52%) of the 21 patients with RTK but none was seen in the comparison group of patients. CONCLUSION: On CT, a peripheral crescent with the attenuation of fluid is characteristic of RTK. However, 12% of renal neoplasms that occur more commonly than RTK in children had CT findings indistinguishable from those of RTK. Because the prevalence of RTK is relatively low, and because the CT findings are not pathognomonic, a renal mass seen on CT in a child is unlikely to represent RTK regardless of its CT features. We therefore conclude that the routine addition of CT of the brain for pediatric patients with renal masses that show a peripheral crescent of fluid attenuation is not justified. Supplemental imaging of the brain should be based on clinical findings or tissue diagnosis.

Brain Neoplasms↗

PubMed1997

The kidney: radiologic-pathologic correlation.

Selected concepts of MR-pathologic correlation of renal masses are presented in this article by way of illustrative examples, with an emphasis on renal adenocarcinoma. Where applicable, practical uses of these concepts are suggested, particularly in those cases in which the differential diagnosis might be narrowed, such as in the evaluation of angiomyolipoma. Also included is a discussion of management decisions that may be affected by the information provided by MR imaging.

Carcinoma, Renal Cell↗

PubMed1997

The adrenal gland: radiologic-pathologic correlation.

The adrenal glands are affected by complex physiologic and neoplastic processes. Recently described computed tomography densitometry and chemical shift MR imaging techniques are useful to distinguish between benign and malignant masses. Knowledge of the basic gross pathologic and histologic correlates of adrenal disease helps in applying these new imaging methods and in understanding established cross-sectional studies.

Adrenal Gland Diseases↗

PubMed1997

Structural determination in AUF1 required for high affinity binding to A + U-rich elements.

AUF1 is a RNA-binding protein that contains two non-identical RNA recognition motifs (RRMs). AUF1 binds to A + U-rich elements (AREs) with high affinity. The binding of AUF1 to AREs is believed to serve as a signal to a mRNA processing pathway which degrades mRNAs encoding many cytokines, oncoproteins and G protein-coupled receptors. Because the ARE-binding activity of AUF1 appears central to the regulation of many important genes, we analyzed the domains of the protein that are important for this activity. Examination of the binding affinity of various mutants indicates that both RRMs may be required for binding. However, they are not sufficient. Highest affinity binding requires an alanine/glycine-rich region of the N-terminus and a short glutamine-rich region in the C-terminus. The N-terminus is required for dimerization of AUF1. However, AUF1 binds an ARE as a hexameric protein. Thus protein-protein interactions are important for high affinity ARE-binding activity of AUF1.

Binding Sites↗

PubMed1996

cDNA cloning of a chick homologue of human ATPase complex subunit 4, quantitative tissue distribution and tertiary structure comparison of the ATPase domain to RecA.

The ocular lens consists of a single layer of epithelial cells on its anterior surface and underlying fiber cells, which are derived from the epithelial cells by differentiation and make up the bulk of the lens. Because lens cells are segregated by age and stage of differentiation, we are using this tissue to study the role of the proteasome in differentiation. The purpose of this study is to corroborate the ATPase function of chick subunit 4 (cS4) and assess the levels of the mRNA in the differentiating lens relative to other tissues. We have generated a computer model of the tertiary structure of the ATPase domain of the cS4 of the ATPase complex that regulates the 20S proteasome. The predicted polypeptide from the cloned cDNA of cS4 (440 residues) had a calculated molecular mass of 49,182 and is 98 and 73% identical to human and yeast S4 protein sequences, respectively. A computer search for comparison with known proteins in GenBank showed that the cS4 protein sequence has a conserved region of about 200 amino acid residues including an ATP/GTP binding site and a mitochondrial energy transfer proteins signature sequence. Based on secondary structure, the computer-generated model of the ATPase domain is comparable to that of RecA, with a root mean square deviation of 0.851 from the RecA triad. mRNA in the 14-day-old chick embryo lens is derived primarily (90%) from differentiating cells. The level of cS4 mRNA determined by quantitative RT/PCR in this differentiating tissue was comparable to the cS4 mRNA levels in chick liver, heart, and brain.

Adenosine Triphosphatases↗

PubMed1996

Localization and physical mapping of genes encoding the A+U-rich element RNA-binding protein AUF1 to human chromosomes 4 and X.

Messenger RNAs encoding many oncoproteins and cytokines are relatively unstable. Their instability, which ensures appropriate levels and timing of expression, is controlled in part by proteins that bind to A+U-rich instability elements (AREs) present in the 3'-untranslated regions of the mRNAs. cDNAs encoding the AUF1 family of ARE-binding proteins were cloned from human and murine cDNA libraries. In the present study monochromosomal somatic cell hybrids were used to localize two AUF1 loci to human chromosomes 4 and X. In situ hybridization analyses using P1 clones as probes identified the 4q21.1-q21.2 and Xq12 regions as the locations of the AUF1 genes.

Animals↗

PubMed1996

Ultrasound versus radiography in the detection of soft-tissue foreign bodies.

STUDY OBJECTIVE: To determine the usefulness of ultrasound and radiography in detecting foreign bodies in soft-tissue models closely duplicating puncture-wound trauma and hand anatomy. METHODS: In this randomized, blinded descriptive study, two radiologists independently evaluated 120 chicken thighs for foreign bodies with the use of standard two-view radiography and 7.5-MHz transducer ultrasonography. All chicken thighs were manipulated with hemostats to ensure uniform tissue damage. In 60 thighs, one foreign body had been inserted (10 each: gravel, metal, glass, cactus spine, wood, and plastic). RESULTS: The sensitivity of ultrasound in detecting gravel was 40%, that for metal was 45%, that for glass was 50%, that for cactus spine was 30%, that for wood was 50%, and that for plastic was 40%. The overall sensitivity, specificity, and false-negative and false-positive rates for ultrasound were 43%, 70%, 50%, and 30%, respectively. No individual foreign body had an ultrasound detection rate of 50%. Radiography detected foreign bodies generally considered radiopaque (gravel, glass, metal) 98% of the time, but it never detected bodies considered radiolucent (wood, plastic, cactus spine). The false-negative and false-positive rates for radiography were 50% and 1.6%, respectively. CONCLUSION: Ultrasound detection of foreign bodies by skilled operators in this animal model revealed poor sensitivity and specificity. Radiographic detection was highly sensitive for foreign bodies considered radiopaque. Our data suggest that ultrasound should not be relied on to rule out the possibility of a retained foreign body in the distal extremities.

Animals↗

PubMed1995

Age-dependent association of isolated bovine lens multicatalytic proteinase complex (proteasome) with heat-shock protein 90, an endogenous inhibitor.

The multicatalytic proteinase complex (MPC) (proteasome) is a high-molecular-weight proteolytic enzyme found in eukaryotic cells and archaebacteria. Regulatory proteins that inhibit or activate the MPC have been described. Association with an ATPase complex alters the specificity of the multicatalytic proteinase complex to permit cleavage of ubiquitinylated proteins. Unidentified proteins have been observed in highly purified preparations of the multicatalytic proteinase complex. Based on immunoreactivity and N-terminal sequencing, we have identified heat-shock protein 90 as a major component of the multicatalytic proteinase complex prepared from 1-month, but not 2-year bovine lenses. alpha-Crystallin, a lens structural protein with chaperone activity, is also found in multicatalytic proteinase complex preparations. Both heat-shock protein 90 and alpha-crystallin inhibit hydrolysis of Cbz-Leu-Leu-Leu-MCA by the multicatalytic proteinase complex at a stoichiometry of 1 mol heat-shock protein per mole of MPC. Heat-shock proteins may interact with denatured proteins and facilitate their degradation. These studies give evidence for the involvement of heat-shock proteins in proteolysis by direct interaction with the multicatalytic proteinase complex.

Age Factors↗

PubMed1995

D20S16 is a complex interspersed repeated sequence: genetic and physical analysis of the locus.

The genomic structure of the D20S16 locus has been evaluated using genetic and physical methods. D20S16, originally detected with the probe CRI-L1214, is a highly informative, complex restriction fragment length polymorphism consisting of two separate allelic systems. The allelic systems have the characteristics of conventional VNTR polymorphisms and are separated by recombination (theta = 0.02, Zmax = 74.82), as demonstrated in family studies. Most of these recombination events are meiotic crossovers and are maternal in origin, but two, including deletion of the locus in a cell line from a CEPH family member, occur without evidence for exchange of flanking markers. DNA sequence analysis suggests that the basis of the polymorphism is variable numbers of a 98-bp sequence tandemly repeated with 87 to 90% sequence similarity between repeats. The 98-bp repeat is a dimer of 49 bp sequence with 45 to 98% identity between the elements. In addition, nonpolymorphic genomic sequences adjacent to the polymorphic 98-bp repeat tracts are also repeated but are not polymorphic, i.e., show no individual to individual variation. Restriction enzyme mapping of cosmids containing the CRI-L1214 sequence suggests that there are multiple interspersed repeats of the CRI-L1214 sequence on chromosome 20. The results of dual-color fluorescence in situ hybridization experiments with interphase nuclei are also consistent with multiple repeats of an interspersed sequence on chromosome 20.

Alleles↗

PubMed1995

Intracranial vascular malformations.

Intracranial vascular malformations are nonneoplastic developmental anomalies that present a variety of clinical patterns, ranging from asymptomatic to fatal intracranial hemorrhage. A practicing radiologist can expect to encounter these abnormalities, which include capillary telangiectasias, venous angiomas (also called developmental venous anomalies), cavernous angiomas, and arteriovenous malformations. The imaging findings that characterize these lesions are reviewed in this article, along with their pathological and clinical features.

Brain↗