Identification of novel MyoD gene targets in proliferating myogenic stem cells.
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A major control point for skeletal myogenesis revolves around the muscle basic helix-loop-helix gene family that includes MyoD, Myf-5, myogenin, and MRF4. Myogenin and MRF4 are thought to be essential to terminal differentiation events, whereas MyoD and Myf-5 are critical to establishing the myogenic cell lineage and producing committed, undifferentiated myogenic stem cells (myoblasts). Although mouse genetic studies have revealed the importance of MyoD and Myf-5 for myoblast development, the genetic targets of MyoD and Myf-5 activity in undifferentiated myoblasts remain unknown. In this study, we investigated the function of MyoD as a transcriptional activator in undifferentiated myoblasts. By using conditional expression of MyoD, in conjunction with suppression subtractive hybridizations, we show that the Id3 and NP1 (neuronal pentraxin 1) genes become transcriptionally active following MyoD induction in undifferentiated myoblasts. Activation of Id3 and NP1 represents a stable, heritable event that does not rely on continued MyoD activity and is not subject to negative regulation by an activated H-Ras G12V protein. These results are the first to demonstrate that MyoD functions as a transcriptional activator in myogenic stem cells and that this key myogenic regulatory factor exhibits different gene target specificities, depending upon the cellular environment. (+info)
Limited sensitivity of parathyroid imaging with (99m)Tc-sestamibi/(123)I subtraction in an endemic goiter area.
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Double-phase single-tracer scintigraphy with (99m)Tc-sestamibi is now generally used for parathyroid imaging but, at least in endemic goiter areas, complementary thyroid scintigraphy is recommended. Although (123)I-sodium iodide is considered to be the optimal thyroid agent, it is hardly ever used because of high costs and logistic difficulties. Our study presents the results of using the (99m)Tc-sestamibi/(123)I subtraction technique in a region with a high goiter prevalence. Special attention was paid to the changes in sensitivity and specificity and their relationship to thyroid volume as well as to autonomous and nodular thyroid disease. METHODS: One hundred three scintigraphic parathyroid examinations on 96 patients were included in this study. Fifty-eight of all patients had concomitant morphologic or functional alterations of the thyroid. Initially, 10 MBq (123)I-sodium iodide were injected. Then, 150 MBq (99m)Tc-sestamibi were administered after 3-5 h, followed by planar scintigraphic imaging of the neck and upper chest region using a double-isotope technique. RESULTS: An area with increased tracer uptake on the subtraction image was found in 44 cases. Forty-three of them proved to be true-positive. No suspicious lesions were detected scintigraphically on the remaining 59 examinations. However, histologic examination revealed a parathyroid adenoma or hyperplasia in 11 of these cases. The mean parathyroid volume of these false-negative patients was 0.9 mL. Secondary hyperparathyroidism with multiple enlarged parathyroid glands was found in 4 of these cases. The sensitivity of the parathyroid scintigraphy was 80% (43/54) and the specificity was 98% (48/49). There was a distinct difference in the sensitivity between the subgroups with thyroid volumes of >15 mL and <15 mL (76% vs. 88%), although the resected parathyroid glands had a similar size in both subgroups. The specificity was 97% and 100%, respectively. No significant difference in the sensitivity and specificity was observed between the subgroups with and without morphologic or functional alterations of the thyroid (80% vs. 79% and 96% vs. 100%, respectively). CONCLUSION: The sensitivity of parathyroid imaging with (99m)Tc-sestamibi/ (123)I subtraction depends mainly on the thyroid and parathyroid volumes rather than on the presence of nodular or autonomous thyroid disease. (+info)
Computer-aided detection and diagnosis at the start of the third millennium.
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Computer-aided diagnosis has been under development for more than 3 decades. The rate of progress appears exponential, with either recent approval or pending approval for devices focusing on mammography, chest radiographs, and chest CT. Related technologies improve diagnosis for many other types of medical images including virtual colonography, vascular imaging, as well as automated quantitation of image-derived metrics. A variety of techniques are currently employed with success, likely reflecting the variety of imagery used, as well as the variety of tasks. Most areas of medical imaging have had efforts at computer assistance, and some have even received FDA approval and can be reimbursed. We anticipate that the rapid advance of these technologies will continue, and that application will broaden to cover much of medical imaging. Acceptance of, and integration of computer-aided diagnosis technology with the electronic radiology practice is a current challenge. These challenges will be overcome, and we expect that computer-aided diagnosis will be routinely applied to medical images. (+info)
Comparative study of MR sialography and digital subtraction sialography for benign salivary gland disorders.
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BACKGROUND AND PURPOSE: MR sialography has become an alternative imaging technique for ductal salivary gland diseases. We compared the diagnostic accuracies of MR sialography and digital subtraction sialography in patients with successful completion of both examinations and benign salivary gland disorders. METHODS: In a prospective study, we attempted to examine salivary glands in 80 patients with clinically suspected diagnoses of sialadenitis and/or sialolithiasis. Each patient underwent digital subtraction sialography and MR sialography. MR sialography was obtained with a T2-weighted single-shot turbo spin-echo sequence (TR/TE 2800/1100 msec, acquisition time 7 seconds), with use of a quadrature head coil. Final diagnoses were confirmed by clinical follow-up and results of biopsy (n = 9) or surgery (n = 19). RESULTS: Failure rate was 5% (four of 80) for MR sialography and 14% (11 of 80) for digital subtraction sialography. Eighty-one salivary glands (48 parotid, 33 submandibular) in 65 patients were successfully visualized with both modalities. MR sialography depicted the main ductal system and first- and second-order branches, whereas digital subtraction sialography was able to depict third-order branches. Sensitivity and specificity to diagnose chronic sialadenitis were 70% and 98% with MR and 96% and 100% with digital subtraction sialography. MR sialography enabled diagnosis of sialolithiasis with a sensitivity of 80% and a specificity of 98% versus 90% and 98% for each with digital subtraction sialography. CONCLUSION: MR sialography with a heavily T2-weighted sequence is highly successful in the noninvasive visualization of the ductal system of major salivary glands. It is useful for diagnosing sialolithiasis and sialadenitis. Digital subtraction sialography, an invasive technique, had a substantial procedural failure rate, particularly for the submandibular duct. However, because of its higher spatial resolution, successfully completed digital subtraction sialography achieved superior diagnostic information compared with that of MR sialography. (+info)
Subtractive hybridization reveals the expression of immunoglobulin-like transcript 7, Eph-B1, granzyme B, and 3 novel transcripts in human plasmacytoid dendritic cells.
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Recent studies in humans have highlighted the importance of a distinct cellular entity, the plasmacytoid dendritic cell (PDC). To identify genes for which expression is restricted to human PDCs, a cDNA subtraction technique was applied using cDNA from activated monocyte-derived DCs (MDDCs) as competitor. In the 650 sequences analyzed, 25% were for B-cell transcripts. We also found lymphoid-related genes, immunoglobulinlike transcript 7 (ILT7), granzyme B (GrB), Spi-B, and the receptor tyrosine kinase Eph-B1. Granzyme B was up-regulated on activation, and protein was detected only in PDCs. Eph-B1 protein was expressed in the cytoplasm and the nuclei of PDCs and MDDCs, respectively. Interestingly, several novel molecules have been identified that were predicted to encode for a type 2 transmembrane protein (BRI(3)), a putative cytokine (C-15, a cysteine-rich-secreted protein), and a type 1 leucine-rich repeat protein (MAPA). The identification of genes expressed in PDCs provides new insights into their function and origin. (+info)
Automated detection of local normalization areas for ictal-interictal subtraction brain SPECT.
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Whole-brain activity is often chosen to quantitatively normalize peri-ictal and interictal SPECT scans before their subtraction. This use is not justified, because significant and extended modification of the cerebral blood flow can occur during a seizure. We validated and compared 2 automatic methods able to determine the optimal reference region, using simulation and clinical data. METHODS: In the first method, the selected reference region is the intersection of peri-ictal-interictal areas with no significantly different z values. The other method relies on a 3-dimensional iterative voxel aggregation. The increase of the selected volume is stopped by using 2 different variance tests (Levene and SE). These algorithms were tested on 39 epileptic patients and were validated using 1 interictal and 10 peri-ictal scans simulated from the mean image of 22 healthy subjects. RESULTS: In the patient studies, the mean relative activity of the selected regions, compared with whole-brain activity (classic normalization), was 122.6%. Their average relative size (compared with the size of the whole brain) was 33.2% for the z map method, 22.8% for the SE test, and 11.8% for the Levene test. After application of our automatic processes, subtraction of the simulated images revealed a recovery of abnormal regions up to 45% larger than the region obtained with classic normalization. CONCLUSION: These results illustrate the role of normalization on the subtracted peri-ictal and interictal images. Our methods are automatic and objective and give good results on various simulated images. The z map construction is worth considering because it is simple, selects large parts of the brain, and requires little computation time. (+info)
The emergence of protocadherin-PC expression during the acquisition of apoptosis-resistance by prostate cancer cells.
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In order to identify gene products associated with the development of acquired therapeutic resistance by prostate cancer cells, we created two novel apoptosis-resistant prostate cancer cell lines, LNCaP-TR (phorbol-ester [TPA]-Resistant) and LNCaP-SSR (Serum Starvation-Resistant) by repeated transient exposure of cultured human LNCaP cells to apoptotic stimuli followed by expansion of surviving cell populations. These cell lines were found to be cross-resistant to the alternative selective agent and also hormone-resistant when xenografted into castrated male immunodeficient mice. RNA from the LNCaP-TR line was comparatively screened using a subtractive hybridization-PCR procedure. This allowed us to identify a 249 bp cDNA fragment that hybridized to a 4.8 kb mRNA preferentially expressed by the apoptosis-resistant cells. Using RACE procedures, we cloned and sequenced the complete 4.8 kb cDNA. It is an unusual member of the protocadherin gene family containing two large overlapping open reading frames encoding homologous polypeptides, one having a signal sequence and the other lacking a signal sequence and we refer to it as protocadherin-PC. LNCaP cells directly transformed with protocadherin-PC cDNA were comparatively resistant to phorbol-ester induced apoptosis. Antibody recognition studies demonstrating the cytoplasmic nature of the protcadherin-PC translation product and its propensity to bind beta-catenin suggest that it might influence the apoptotic sensitivity of prostate cancer cells through a unique mechanism. (+info)
A novel nuclear zinc finger protein EZI enhances nuclear retention and transactivation of STAT3.
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A novel cDNA EZI isolated as an oncostatin M- inducible gene encoded a protein containing 12 C2H2-type zinc fingers. EZI was found to transactivate the promoters that are also responsive to STAT3 and activated the acute phase response element (APRE) synergistically with STAT3. Co-immunoprecipitation demonstrated the association of EZI with STAT3, which was mediated by the N-terminal region (1-183) of EZI. The EZI mutant lacking this region showed reduced transcriptional activity, indicating that EZI and STAT3 function cooperatively through physical interaction. While EZI predominantly localized in the nucleus and enhanced the nuclear localization of STAT3, the EZI mutant lacking 11 zinc finger motifs failed to translocate into the nucleus and also inhibited nuclear localization of STAT3 as well as STAT3-mediated transactivation. These results indicate that EZI is a novel nuclear zinc finger protein that augments STAT3 activity by keeping it in the nucleus. (+info)