Search PubMed⌕ Search

Biomedical subjects

T R Brown

Publications and source records attributed to T R Brown.

At least 37 records · Page 2Linked to original sources
PubMed1998

Telomerase activity in normal adult Brown Norway rat seminal vesicle: regional distribution and age-dependent changes.

Telomerase activity is essential for protection of cells against the telomere erosion that occurs with each round of cell replication, and thus appears to play a role in the indefinite replication potential of some, but not all, eukaryotic cells. In this regard, some tissues contain stem cells that have a long proliferative life-span and are capable of regenerating or renewing the somatic epithelial cell population within the tissue. Because the adult seminal vesicle exhibits the ability to regenerate during androgen-replacement after castration, we hypothesized that a pool of cells with regenerating potential is present in the adult seminal vesicle, which expresses telomerase activity. In this study, we used a highly sensitive PCR-based telomerase assay [the telomeric repeat amplification protocol (TRAP) assay] to detect telomerase activity in rat seminal vesicle. Our results show that telomerase activity is, indeed, present in the normal adult rat seminal vesicle, but that, in the presence of seminal vesicle fluid, telomerase activity cannot be detected. In fact, seminal vesicle fluid was found to contain some factor(s) that is inhibitory for the TRAP assay. In addition, we found that telomerase activity in the seminal vesicle changes with age and is regionally distributed within the distal, intermediate, and proximal segments of the duct. These results suggest that as is the case for the rat prostate, a population of telomerase-positive cells is present within the adult rat seminal vesicle, and thereby, this organ retains throughout life the potential to regenerate in response to androgen replacement following castration-induced apoptotic cell death.

Age Factors↗
PubMed1998

Regional expression of transforming growth factor-alpha in rat ventral prostate during postnatal development, after androgen ablation, and after androgen replacement.

The prostate is a highly heterogeneous organ, composed of different types of epithelial and stromal cells organized regionally along the ductal network. Although androgen-stimulated growth and maintenance of the prostate gland primarily involve epithelial cells, it is unclear whether all epithelial cells are androgen dependent. Moreover, the actions of androgens may not be direct; a number of polypeptide growth factors, including transforming growth factor-alpha (TGFalpha), are postulated to mediate androgen action in the rat prostate. In this investigation, using an immunohistochemical technique, we examined the cellular and regional expression of TGFalpha in the rat ventral prostate during postnatal development to adulthood. TGFalpha-immunopositive cells were located throughout the ductal epithelium from postnatal days 5-20. By day 45 and thereafter, regional variation in TGFalpha expression became apparent; epithelial cells in the proximal segment exhibited intense staining, whereas those in the distal segment exhibited negligible staining. These observations were coincident with increased serum testosterone concentrations at puberty. To understand the role of androgen in the expression of TGFalpha in the epithelial cells of the distal and proximal segments of the adult rat ventral prostate, androgen was withdrawn by castration, and testosterone subsequently was administered. Androgen receptor protein expression decreased after castration and reappeared after androgen replacement in both the distal and proximal segments. TGFalpha staining was negligible in epithelial cells of the distal segment of intact adult rats, became prominent by 7 days after castration, but then diminished after the administration of testosterone. Western blot analyses revealed the presence of a specific 30-kDa immunoreactive form of TGFalpha in rat ventral prostate, and its quantity reflected the staining intensities observed in the immunohistochemical studies. These results suggest that TGFalpha expression is negatively regulated by androgen in epithelial cells of the distal segment. In contrast, staining for TGFalpha in epithelial cells of the proximal segment did not change with castration or testosterone administration, suggesting that TGFalpha is not regulated by androgen in this region of the ventral prostate. In summary, TGFalpha expression is differentially regulated among epithelial cells localized in two different regions of the ventral prostate. We hypothesize that TGFalpha may function as a survival factor for epithelial cells which, as a consequence of its expression, become androgen independent and thus escape apoptotic cell death after androgen ablation.

Aging↗
PubMed1998

Thyroid injury, autoantigen availability, and the initiation of autoimmune thyroiditis.

Iodine depletion prevents disease and iodine repletion, which may cause thyroid cell injury by reactive oxygen intermediates, initiates disease in the Obese Strain chicken model of spontaneous autoimmune thyroiditis (AT). To examine the role of cell injury and autoantigen availability in AT induction we compared the immune responses that followed blunt trauma to the OS thyroid in the absence of iodine and the administration of normal dietary iodine in the absence of thyroid injury. Serum thyroglobulin concentrations were elevated following thyroid injury and the extensive thyroid infiltrates had high macrophage/CD4+, CD8+, B cell ratios consistent with an acute inflammatory response. The response was self limiting and undetectable in all animals 2 weeks later. Birds raised on a similar low iodine regimen were withdrawn from the regimen and given normal dietary iodine. Their thyroids showed no evidence of acute ultrastructural damage. The resulting early thyroid infiltrates had low macrophage/CD4+, CD8+, B cell ratios. Two weeks later these animals showed severe thyroid infiltration (43%) and after 4 weeks all animals had >90% infiltration. Thus, the presence or absence of thyroidal iodine, whether accompanied by injury or not, determined the nature and consequence of the immune response which argues against hypotheses that include obligatory injury at disease onset. Taken with previous work, this study suggests that iodination of an autoreactive thyroglobulin epitope is a requisite pathogenic action of iodine and supports the notion that in some organ-specific autoimmunity, a component of the dysregulation is associated with aberrant activity of the target tissue.

Age Factors↗
PubMed1997

Stable transfection of U937 cells with sense or antisense RXR-alpha cDNA suggests a role for RXR-alpha in the control of monoblastic differentiation induced by retinoic acid and vitamin D.

Although retinoic acid (RA) has been known for many years to be a modulating agent that plays a role in generating both granulocytes and monocytes, the molecular mechanism underlying this role has not been defined in the monoblast lineage. In particular, the part played by the retinoid X receptors (RXRs), which are members of the steroid/thyroid hormone nuclear receptor family, has not been explored. In this study, therefore, the human monoblastic leukemia cell line U937 has been used as a model system to investigate the role of one of the RXRs, RXR-alpha, in monoblast differentiation. RXR-alpha mRNA was present in untreated U937 cells, and levels increased after induction of differentiation with phorbol ester. The same was found for RXR-beta mRNA. Using plasmids containing sense or antisense RXR-alpha sequences under the control of an inducible promoter, we generated stably transfected cell lines which expressed either increased or decreased levels of RXR-alpha, respectively. The sense cell lines (U alpha S and its clonal derivative alpha G2S) showed increased sensitivity to RA, while the antisense cell lines (U alpha A and its clonal derivative alpha B5A) showed decreased sensitivity to RA, as demonstrated by growth inhibition and by regulation of an RA-responsive reporter gene. Both U alpha A and alpha B5A also failed to respond to another modulating agent, 1 alpha,25-dihydroxycholecalciferol (DHCC), but only U alpha S and not alpha G2S showed an enhanced response to DHCC. The combination of RA and DHCC together inhibited growth of both sense and antisense cell lines. In addition, alpha G2S exhibited increased expression of CD11b and CD54, while alpha B5A cells showed increased expression of CD102, suggesting that RXR-alpha has a role in regulating expression of cell adhesion molecules in U937 cells. These results demonstrate that RXR-alpha has a role in mediating growth inhibition and cell adhesion during myelomonocytic differentiation, and suggest that different species of heterodimers involving RXR-alpha may control the acquisition of different features of mature monocyte/macrophage function.

Antigens, CD↗
PubMed1997

Quantitation of 3-deoxyglucosone levels in human plasma.

3-Deoxyglucosone (3DG), a reactive dicarbonyl, is an important intermediate in the formation of advanced glycation end products (AGEs). The AGEs are particularly important in diabetes since they have been correlated with the development of diabetic complications. Consequently, measurements of 3DG are likely to provide valuable insights into the role of this metabolite in the etiology of diabetic complications. While several methods of 3DG quantitation in human plasma have been previously published, a significant discrepancy (over 30-fold) exists in the reported values. Knecht et al. (Arch. Biochem. Biophys. 294, 130-137, 1992) have reported the levels of plasma 3DG in normoglycemics to be 61 nM, using a GC/MS procedure. In contrast to this, Niwa et al. (Biochem. Biophys. Res. Commun. 196, 837-843, 1993) reported 3DG levels to be 1800 nM in normoglycemics, using a totally independent GC/MS method. To resolve this disagreement and fill the need for a robust assay for this dicarbonyl, suitable for absolute quantitation, a GC/MS procedure was devised for its measurement. Plasma samples were deproteinized either by ultrafiltration or by addition of ethanol as described by Niwa et al. (Biochem. Biophys. Res. Commun. 196, 837-843, 1993). 3DG in the ultrafiltrate or the supernatant was conjugated with 2,3-diamino-naphthalene to produce a stable adduct which was then converted to a silyl ether and analyzed by GC/MS. The analyte was monitored by selected ion monitoring at an m/z of 295 and 306 and quantitated using an internal standard of [U-13C]3DG. Using this approach, 3DG levels in plasma deproteinized by ultrafiltration were found to be significantly elevated from 58.5 +/- 14 (SD) nM in normoglycemics to 98.5 +/- 34 (SD) nM in type I diabetics. When deproteinization of the plasma was carried out using ethanol, the levels of 3DG from normoglycemic plasma were similar to those reported by Niwa et al. (1710 +/- 750 (SD) nM). These results suggest that 3DG levels measured by ultrafiltration may represent the free circulating 3DG and those obtained by ethanol extraction may represent aform of 3DG bound to a macromolecule (presumbaly protein).

2-Naphthylamine↗
PubMed1997

Identification and characterization of a novel androgen response element composed of a direct repeat.

Transcriptional regulation by the androgen receptor (AR) requires its binding to hormone response element nucleotide sequences in DNA. A consensus glucocorticoid response element (GRE) can mediate transactivation by AR and other members of the AR/glucocorticoid (GR)/progesterone (PR)/mineralocorticoid (MR) receptor subfamily. We identified putative androgen response element (ARE) sequences by binding of a human AR DNA-binding domain fusion protein to DNA in a random sequence selection assay. A 17-base pair consensus nucleotide sequence, termed IDR17, containing three potential GRE-like core binding sites organized as both inverted and direct repeats, was determined from a pool of degenerate oligonucleotides. IDR17 was active in mediating androgen-dependent induction of reporter gene expression in transient transfection assays. Dissection of the IDR17 sequence revealed an 11-base pair sequence (DR-1), consisting of two potential core binding sites oriented as an overlapping direct repeat, as the most potent ARE. DR-1 demonstrated a strong preference for AR binding and transactivation when compared with GR. To our knowledge, this is the first observation that a direct repeat of GRE-like core motifs functions as a preferred hormone response element within the AR/GR/PR/MR subfamily of nuclear receptors.

Base Sequence↗
PubMed1997

Simultaneous 3D NMR spectroscopy of proton-decoupled fluorine and phosphorus in human liver during 5-fluorouracil chemotherapy.

Simultaneous acquisition of 1H-decoupled 31P and 19F 3D CSI is demonstrated in the liver of a patient undergoing 5-fluorouracil chemotherapy. Both 31P and 19F shared the same voxel size (64 or 27 ml), bi-level 1H-decoupling and 0.35 s TR. The measurements were done in a 1.5 Tesla clinical imager with three radio-frequency (RF) channels and a triple-tuned surface-coil. The overall MRI and MRS examination time was under 90 min. Simultaneous acquisition of 31P and 19F permits localized study of the influence of hepatic metabolism on the uptake and catabolism of fluoropyrimidine drugs without extra measurement time or higher SAR.

Absorption↗
PubMed1997

In vivo phosphorus polarization transfer and decoupling from protons in three-dimensional localized nuclear magnetic resonance spectroscopy of human brain.

Refocused insensitive nucleus enhancement by polarization transfer (RINEPT) from protons (1H) to a J-coupled phosphorus (31P) has been incorporated into three-dimensional (3D) chemical-shift-imaging (CSI) sequence on a clinical imager. The technique is demonstrated on a phantom and in in vivo human brain. The polarization-transfer efficiency (approximately 1.2) is lower than the theoretical maximum of gamma1H/gamma31P approximately 2.4 resulting from 1H-1H homonuclear J couplings of similar magnitude competing with the 1H --> 31P transfer. Nevertheless, compared with direct 31P Ernst-angle excitation, signal gains of up to x1.8 were obtained mainly as a result of T1 differences between 31P and the 1H. Spectral interpretation is simplified by editing out all non-proton-coupled 31P signals. The duration, approximately 50 min, and power deposition, approximately 1 W x kg(-1), make the application suitable for human studies.

Brain↗
PubMed1997

Proton-decoupled 19F spectroscopy of 5-FU catabolites in human liver.

An RF network and a dual-tuned surface coil are described for obtaining proton-decoupled, NOE enhanced 19F spectra from a whole body clinical imager operating at 1.5 Tesia. The network removes 19F frequency noise from the decoupler transmitter, and prevents preamplifier saturation from high-level decoupling signals. Proton decoupling of 19F spectra was optimized using a sample of urine containing 5-fluorouracil (5-FU) and its catabolite fluoro-beta-alanine (FBAL). Proton-decoupled 19F spectroscopy in vivo is demonstrated by obtaining both nonlocalized spectra and spectra localized with three-dimensional chemical shift imaging from the liver of patients undergoing 5-FU chemotherapy.

Fluorouracil↗
PubMed1997

Study of the metabolism of choline and phosphatidylcholine in tumors in vivo using phosphonium-choline.

The results of an initial study on the feasibility of using the phosphonium analog of choline to follow the metabolism of phosphatidylcholine in tumors in vivo using 31P NMR are reported. C3H/He mice bearing a mammary carcinoma tumor on the foot pad were fed a choline-free diet supplemented with the phosphonium analog of choline. Metabolites of this compound, including the phosphonium analogs of phosphatidylcholine, phosphocholine, glycerophosphocholine, and betaine were observed noninvasively in vivo in tumors by 31P NMR after 2-3 weeks of feeding. Clearance of these phosphonium-labeled metabolites from tumors was measured after a change to a choline-containing diet. Significant decreases were seen in the levels of the analogs of betaine (P < 0.003) and phosphatidylcholine (P < 0.004) by Day 4. A significant increase in the level of authentic phosphocholine (P < 0.003) occurred over the same time period.

Adenocarcinoma↗
PubMed1997

Fructose-3-phosphate production and polyol pathway metabolism in diabetic rat hearts.

Previous studies have suggested that polyol-pathway and nonenzymatic glycation may be involved in the development of cardiac myopathy, a well-known manifestation of diabetes. Although the exact etiology of this complication is not fully understood, it is likely to be multifactorial. In this study, we investigated the metabolic consequences of diabetes and the effect of aldose reductase inhibitor (ARI) treatment on cardiac tissues of Sprague-Dawley rats. Perchloric acid (PCA) extracts of hearts from the animals were examined using 31P-nuclear magnetic resonance (NMR), gas chromatography/mass spectrometry (GC/MS), and high-performance liquid chromatography (HPLC). In 31P-NMR spectra of diabetic animals, a peak resonating at the chemical shift of 5.8 ppm with a coupling constant of 10 Hz was identified as fructose-3-phosphate (F3P). Undetectable in controls (< approximately 20 nmol/g), this metabolite was present at a concentration of 81.3 +/- 16.3 nmol/g wet weight (n = 4) in diabetic rat hearts. GC/MS analysis of these extracts from diabetics also identified a decomposition product of F3P, 3-deoxyglucosone (3DG), at a concentration of 9.4 +/- 3.5 nmol/g (n = 3), compared with 0.98 +/- 0.43 nmol/g (n = 3) in controls. No evidence was found for the expected detoxification products of 3-DG, 3-deoxyfructose and 2-keto 3-deoxygluconate. Concomitant with the elevation of F3P and 3DG, fructose and sorbitol levels were also elevated in diabetic animals. Surprisingly, ARI treatment was found to have no effect on the levels of these metabolites. These data suggest that either the heart may be unique in its production of fructose or it may not readily transport the ARI sorbinil. Production of the potent glycating agents F3P and 3DG in diabetics suggests that these compounds may be contributing factors in the glycation of cardiac proteins in the diabetic rat heart.

Administration, Oral↗
PubMed1997

Androgen insensitivity with mental retardation: a contiguous gene syndrome?

We present data to suggest the existence of a mental retardation (MR) locus at Xq11.2-q12 between DXS1 and DXS905, identified in two subjects with complete androgen insensitivity syndrome (CAIS) and MR. Androgen insensitivity syndrome is a disorder of male sexual differentiation caused by a defect in the androgen receptor (AR) gene (Xq11-q12). Two subjects with CAIS resulting from a complete deletion of the AR gene have previously been reported, one of whom also has MR. We have identified another mentally retarded person with a complete deletion of the AR gene. The deletion in the two patients with CAIS and MR extends past the AR gene and includes several marker loci both proximal and distal to the AR gene, the limits of the deletions being DXS1 and DXS905. The deletions in the CAIS patients who do not have MR do not include any of the markers outside the AR gene itself. These data suggest that located close to the AR gene is a gene which is implicated in non-specific MR.

Androgens↗
PubMed1997

MR imaging of the rotator cuff tendon: interobserver agreement and analysis of interpretive errors.

PURPOSE: To evaluate accuracy in interpretation of shoulder magnetic resonance (MR) images and interobserver agreement and to characterize the types of errors. MATERIALS AND METHODS: Five radiologists with varying experience independently and retrospectively twice interpreted the MR images of 222 symptomatic patients who underwent both MR imaging and shoulder arthroscopy. The first interpretation was a blind review; the second was with knowledge of the arthroscopic findings. RESULTS: For full-thickness tears, the sensitivity, specificity, and accuracy were 84%-96%, 94%-98%, and 92%-97%, respectively, and for partial tears, 35%-44%, 85%-97%, and 77%-87%, respectively. There was no statistically significant difference between readers in the detection of partial or full-thickness tears. There was a statistically significant difference between readers in the no-tear category. Cohen kappa values generally indicated improved interobserver agreement proportional to the readers' experience (full-thickness tears, 0.731-0.881; partial tears, 0.168-0.443). CONCLUSION: Diagnosis of a full-thickness tear can be learned to a high degree of accuracy. Despite the radiologist's level of experience and knowledge of the arthroscopic findings, the sensitivity for diagnosis of partial tears is poor.

Adolescent↗
PubMed1996

Metabolic characterization of human soft tissue sarcomas in vivo and in vitro using proton-decoupled phosphorus magnetic resonance spectroscopy.

We applied 1H-decoupling and nuclear Overhauser enhancement to obtain well-resolved 31P magnetic resonance spectra accurately localized to 20 soft tissue sarcomas in vivo, using three-dimensional chemical shift imaging. Fifteen spectra had large phosphomonoester signals (21% of total phosphorus) that contained high amounts of phosphoethanolamine (compared to those of phosphocholine) but no signals from glycerophosphoethanolamine, and glycerophosphocholine was detected in only four cases. Prominent nucleoside triphosphates (52% of phosphorus) and low inorganic phosphate (10% of phosphorus) indicated that a large fraction of these 15 sarcomas contained viable cells, and this impression was confirmed histologically in 13 of the sarcomas. High-resolution in vitro 31P spectra of extracts of surgical specimens of four of the sarcomas studied in vivo and six additional sarcomas confirmed the in vivo assignments of metabolites and revealed considerable inter- and intratumoral variations of metabolite concentrations associated with histological variations in the relative amounts of cells and of matrix materials or spontaneous necrosis. Seven sarcomas, all high grade with pleomorphic or round cells rather than spindle cells, contained an unidentified phosphodiester signal in vivo; its absence in the extract spectra indicates that it may be from an abnormally mobile membrane component. We have documented a means to obtain new information about in vivo metabolism in human sarcomas and to provide a basis on which to examine the uses of 31P magnetic resonance spectroscopy in the clinical management of sarcomas.

Adult↗
PubMed1996

Molecular modeling and in vitro investigations of the human androgen receptor DNA-binding domain: application for the study of two mutations.

In two families with complete androgen insensitivity, we have identified naturally occurring point mutations in the human androgen receptor gene that encode amino acid substitutions within the DNA-binding domain. The two amino acid substitutions, a valine to phenylalanine and a leucine to proline, occur at positions 581 and 616, respectively, of the androgen receptor. The mutations were recreated by site-directed mutagenesis. Expression of the mutants androgen receptors in COS 7 and CV 1 cells revealed a normal size 110-kDa receptor protein on Western blots, normal androgen binding capacities and affinities, but absence of binding to target DNA on electrophoretic mobility shift assays. In cotransfection assays, the mutant ARs failed to activate transcription of an androgen-responsive reporter gene. To study the possible structural impact of these mutations, three-dimensional models of the normal androgen receptor and of each mutant were built by homology with the glucocorticoid receptor. Analysis of the models predicts that mutation Val581Phe would affect interaction between the protein and DNA, whereas the Leu616Pro mutation would more likely be involved in destabilizing the protein structure or protein dimerization. Taken together, the experimental investigations and the molecular modeling studies of the mutant androgen receptors observed in families with complete androgen insensitivity syndrome, highlight the role of Val-581 and Leu-616 in receptor structure and function.

Base Sequence↗
PubMed1996

Heteronuclear multivoxel spectroscopy of in vivo human brain: two-dimensional proton interleaved with three-dimensional 1H-decoupled phosphorus chemical shift imaging.

Multivoxel, heteronuclear interleaved two-dimensional proton and three-dimensional 1H-decoupled 31P CSI of human brain is demonstrated. This method offers efficient use of limited examination time as patient preparation, coil tuning, shimming and imaging are done only once and the CSI data sets from both nuclei are obtained concurrently. Effective interleaving of 31P and 1H is possible due to the shorter T1s of proton brain metabolites, allowing a 1H acquisition cycle to be inserted into each 31P TR. This way, the entire MRS time is available to both nuclei, increasing their SNR per-unit-time by approximately 12% for 31P and approximately 80% for 1H, compared with sequential detection of equal (45-50 min) length. The spectral resolution and SNR of 31P are further increased through bi-level 1H-decoupling and NOE.

Brain↗
PubMed1996

Quantitation of resonances in biological 31P NMR spectra via principal component analysis: potential and limitations.

This paper examines the potential and limitations of peak area quantitation of biological NMR spectra using principal component analysis (PCA), including its requirement for prior knowledge. The principles of the method are presented without in-depth mathematical treatment. PCA is illustrated for simulated data, 31P NMR spectra obtained consecutively over 1-2.5 days from perfused Rat-2 cells metabolizing the choline analogue phosphoniumcholine (Chop) and in vivo proton-decoupled, NOE-enhanced, three-dimensional CSI localized 31P NMR spectra of the liver of healthy volunteers. The results show that PCA can be used to quantitate strongly overlapping peaks without prior knowledge of the peak shapes or positions and to reconstruct spectra with significantly reduced noise variance. Two major limitations of PCA are presented: (1) PCA cannot separate peaks whose intensities are well correlated; (2) PCA is sensitive to differences in chemical shift and line-width of peaks between spectra. The discussion focuses on what knowledge of the biological and spectroscopic features of the samples and the principles of PCA is necessary for peak area quantitation via PCA.

Humans↗