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[Early diagnosis of small hepatocellular carcinoma by new contrast-enhanced ultrasound technique].

OBJECTIVE: To investigate the diagnostic value of contrast-enhanced ultrasound (CEUS) enhancement patterns of hepatocellular carcinoma (HCC) smaller than 2 cm. METHODS: From 392 cases with focal liver lesions undergoing CEUS with SonoVue and Contrast Tuned Imaging (CnTI) technique, thirty six cirrhotic patients with thirty eight HCCs with histopathological diagnosis were retrospectively analyzed in this study. Surgery or needle biopsy was performed within fifteen days after CEUS. Twenty two tumors were finally diagnosed as moderately differentiated HCC, twelve as well differentiated HCC, and four as clear cell carcinomas using Edmondson Grade system. The enhancement patterns of different histopathological HCCs were analyzed. RESULTS: Before CEUS, only 16 lesions (42.1%) were diagnosed as malignant or possibly malignant. After CEUS, 38 HCCs were enhanced in different degrees following contrast administration,of which, all the 22 moderately differentiated HCCs presented typical "fast-in and fast-out" pattern with fast enhancement in arterial phase and then a quick wash-out in the parenchymal phase. Nine (75.0%) of the 12 well differentiated HCCs presented "fast-in and slow-out" pattern. One of the 4 clear cell carcinomas presented "fast-in and fast-out", one presented "fast-in and slow-out" and the other 2 presented "slightly slow-in and slow-out" pattern. According to the enhancement patterns, 76.3% (29 tumors) were considered as malignant, while 18.4% (7 tumors) as possibly malignant, only 5.3% (2 tumors smaller than 1.5 cm) could not be diagnosed by CEUS. CONCLUSION: The identification of different enhancement patterns of various histological types of small HCCs could improve the diagnostic ability of ultrasound in cirrhotic liver. CEUS might be also helpful in detecting small HCCs. Thus CEUS could be a complementary method of enhanced CT and other imaging methods in early diagnosis of HCCs.

Adolescent↗

Value of subtraction images in the detection of hemorrhagic brain lesions on contrast-enhanced MR images.

Contrast-enhanced T1-weighted MR images are known to improve the detection and conspicuity of CNS lesions; however, lesion enhancement may be indistinguishable from contiguous hemorrhagic areas of intrinsically increased signal intensity. In a series of cranial MR studies in 22 pediatric oncologic patients with evidence of subacute hemorrhage, we found that subtracting the unenhanced from the enhanced T1-weighted images was essential in 14 of the cases to visualize and/or characterize the enhanced tissue in the presence of adjacent hemorrhage. In six patients, a nodular pattern of enhancement suggestive of tumor was detected and/or outlined only after subtraction. In eight patients, linear benign-appearing enhancing margins were seen clearly only after subtraction. In the remaining eight patients, the presence and shape of enhancement was verified with subtraction. Subtraction is a useful, simple, and rapid postprocessing procedure that does not increase scan time or require modification of standard pulse sequences. Subtraction separates the enhanced areas from adjacent bright hemorrhage.

Adolescent↗

The enhancer of the human transferrin gene is organized in two structural and functional domains.

We previously identified a 300-base pair long enhancer, located 3.6 kilobases upstream of the cap site of the human transferrin gene. A 5' deletion up to position 86 of the enhancer resulted in complete loss of the enhancer activity. Here we show by competition footprint analysis, gel retardation assays, and transient expression studies in hepatoma and HeLa cells that the enhancer is composed of two distinct structural and functional domains, A (nucleotides 1-86) and B (nucleotides 87-291). Each domain is a proto-enhancer of a different type. Domain A is a proto-enhancer that, when multimerized, is able by itself to stimulate transcription from the heterologous SV40 promoter, both in Hep3B and HeLa cells. It contains the octanucleotide TGTTTGCT sequence and is the binding site of two liver-specific nuclear factors and of a different HeLa nuclear factor. Domain B contains four binding sites interacting with several liver nuclear proteins. In order to bind, any of these proteins requires the presence of all the others. This domain is able to block the activity of a downstream negative element, but it has no enhancer activity by itself. In the presence of the transferrin promoter, full enhancer activity requires the association of the two domains A and B.

Base Sequence↗

Characterization of a human apolipoprotein E gene enhancer element and its associated protein factors.

An enhancer element in the 5' flanking region of the human apolipoprotein E gene, known as upstream regulatory element 1 (URE1), has previously been implicated in the expression of this gene. The URE1 element, which spans nucleotides -193 to -124 of the 5'-flanking region of the human apolipoprotein E gene, contains two sequences that bind to nuclear proteins, as determined by the DNase I footprinting assay. In the present study of URE1, we have characterized these sequences further. Deletion of one of the footprint sequences, at nucleotides -161 to -141, reduced URE1 enhancer activity substantially. A 30-base pair oligonucleotide that included this protein-binding sequence was able, by itself, to act as an enhancer. This sequence, termed the positive element for transcription (PET), was demonstrated by gel retention analysis to bind at least two protein factors, one of which is the transcription factor Sp1. Sp1 appeared to be the only protein required for the enhancer activity of PET to be manifested. In vitro transcription assays showed that the PET sequence was necessary for efficient transcription directed by the apoE promoter and that the PET sequence was the dominant regulatory element in the apoE promoter. Gel filtration chromatography and PET oligonucleotide-affinity chromatography were used to isolate a second PET-binding factor, a Mr = 55,000 protein, from HeLa cell nuclear extracts. It appeared to compete with Sp1 for a common binding site in the PET sequence, but it was not required for enhancer activity. The second footprint sequence in URE1, at nucleotides -184 to -173, also bound Sp1, but it was not required for enhancer activity. A third Sp1-binding region was located at a proximal GC box element (nucleotides -54 to -45). This region had no enhancer activity, but it was required for maximum transcriptional activity of the apoE promoter. Thus, the regulation of apoE gene expression is influenced by different protein-binding sequences, with transcription factor Sp1 playing major roles in both basal promoter activity and enhancer activity.

Animals↗

Cooperative function of two separate enhancers of the human epidermal growth factor receptor proto-oncogene.

Two separate enhancers were identified both upstream and downstream of the promoter of the human epidermal growth factor receptor gene. Transcriptional enhancer activity was found to be associated with a region 1172 and 852 base pairs upstream of the most upstream RNA start site, and within a 530-base pair fragment located in the first intron about 2000 base pairs downstream of the most upstream RNA start site. These two separate enhancers stimulated promoter activity cooperatively in HeLa cells synthesizing the epidermal growth factor receptor. The enhancer downstream of the promoter functioned only in the presence of the other upstream enhancer. Several areas of sequences homology with viral and cellular enhancers were noted in both the upstream and downstream enhancers. The region essential for the downstream enhancer activity has 10 nuclear factor-binding sites.

Base Sequence↗

[Prostate and prostatic carcinoma: comparison of gadolinium-enhanced MR images and histopathologic findings].

The purpose of this study was to determine the patterns of enhancement in normal prostate, and to compare them with the enhancement patterns and histopathologic findings in patients with prostatic carcinoma. Seventeen patients who had no urogenital disease and 27 who underwent radical prostatectomy or prostatic biopsy were included in this study. All patients were evaluated with T1-, T2-weighted and gadolinium-enhanced T1-weighted images. With gadolinium enhancement the inner gland showed more enhancement than the peripheral zone in the normal prostate. The carcinoma tissue was enhanced more than the peripheral zone in 13 patients, about the same as the peripheral zone in 11 patients, and less than the peripheral zone in three patients. The three patients whose cancer tissue demonstrated no enhancement had poorly differentiated adenocarcinoma of the solid type. In conclusion, gadolinium-enhanced T1-weighted imaging is not reliable for routine use in MR imaging of the prostate, but may be useful to evaluate histologic types in patients with prostatic carcinoma.

Adolescent↗

Effect of dose and field strength on enhancement with paramagnetic contrast media.

PURPOSE: To compare contrast enhancement per unit of dose of contrast medium in MR imaging at 0.5 and 1.5 T. METHODS: Contrast enhancement in images made at 0.5 and 1.5 T after 0.1 mmol/kg of gadopentetate dimeglumine and 0.3 mmol/kg of gadodiamide was measured and the degree of contrast enhancement in the cavernous sinus and pituitary gland compared. RESULTS: At both field strengths and both contrast medium doses, contrast enhancement was noted in the cavernous sinus, pituitary gland, infundibulum, maxillary sinus mucosa, falx cerebri, and choroid plexus on inspection of images. Enhancement was significantly and conspicuously less in the cavernous sinus and pituitary gland at 0.5 T (96% and 33%, respectively) than at 1.5 T (160% and 102%, respectively). No tissues were identified that enhanced only with the larger dose or higher field strength. CONCLUSION: In tissues that normally enhance after intravenous administration of gadolinium chelates, enhancement is greater at 1.5 T than at 0.5 T.

Brain Neoplasms↗

Magnetization transfer effects in MR-detected multiple sclerosis lesions: comparison with gadolinium-enhanced spin-echo images and nonenhanced T1-weighted images.

PURPOSE: To define the relationship between magnetization transfer and blood-brain-barrier breakdown in multiple sclerosis lesions using gadolinium enhancement as an index of the latter. METHODS: Two hundred twenty lesions (high-signal abnormalities on T2-weighted images) in 35 multiple sclerosis patients were studied with gadolinium-enhanced spin-echo imaging and magnetization transfer. Lesions were divided into groups having nodular or uniform enhancement, ring enhancement, or no enhancement after gadolinium administration. For 133 lesions, T1-weighted images without contrast enhancement were also analyzed. These lesions were categorized as isointense or hypointense based on their appearance on the unenhanced T1-weighted images. RESULTS: There was no difference between the magnetization transfer ratio (MTR) of lesions as a function of enhancement. MTR of hypointense lesions on unenhanced T1-weighted images was, however, lower than the MTR of isointense lesions. CONCLUSION: We speculate that diminished MTR may reflect diminished myelin content and that hypointensity on T1-weighted images corresponds to demyelination. Central regions of ring-enhancing lesions had a lower MTR than the periphery, suggesting that demyelination in multiple sclerosis lesions occurs centrifugally. In addition, the short-repetition-time pulse sequence seems useful in the evaluation of myelin loss in patients with multiple sclerosis.

Adolescent↗

Combinatorial structure of a body muscle-specific transcriptional enhancer in Caenorhabditis elegans.

We describe the dissection of a body muscle-specific enhancer sequence contained within the Caenorhabditis elegans myosin heavy chain gene unc-54. A 90-base pair segment that was sufficient for both enhancer function and tissue specificity was subjected to mutational analysis. Several separated sites within this region were required for activity; mutations in these sites led to dramatic decreases in enhancer activity, while substitutions in the intervening regions had minimal effects on activity. The individual sites appear to function as semi-independent and partially interchangeable enhancer subelements, as seen by our ability to create functional enhancers by constructing novel multimers and combinations. Four different enhancer subelements (designated O, I, II, and III) were identified in this way. Although partially interchangeable, some differences between these subelements were evident. In particular, concatamers of site III exhibited the highest levels of activity but had a broader tissue specificity than the intact enhancer, including both hypodermal and muscle tissue. The specificity of the intact enhancer thus reflects a combinatorial function of the specificities of the constituent subelements.

Animals↗

Organization of the transcription factor binding sites in the kappa Ig intron enhancer. Effects of position, orientation, and spacing.

The kappa Ig intron enhancer is comprised of multiple sequence motifs known to bind trans-acting factors that activate gene expression. A species comparison reveals a high level of conservation of the organization of the transcription factor binding sites within the enhancer. The importance of the conserved organization of the kappa intron enhancer was examined by using topologic mutations that disrupt the position, orientation, and spacing of individual binding sites within the enhancer. The effects of these changes were monitored by their effects on reporter gene activity at two distinct stages of B cell development. Previously, mutational analysis indicated the kappa B and kappa E2 sequence motifs to be the most crucial sites for intron enhancer function. We have demonstrated that intron enhancer activity is dependent on the position of the kappa B and kappa E2 sequence motifs within the enhancer. Intron enhancer function is, however, independent of kappa B and kappa E2 binding site orientation and is flexible in spacing requirements among binding sites.

Animals↗

The murine Ig 3' alpha enhancer is a target site with repressor function for the B cell lineage-specific transcription factor BSAP (NF-HB, S alpha-BP).

Using electrophoretic mobility shift assays (EMSAs) we have identified several target sites for nuclear proteins in the murine heavy chain Ig 3' alpha enhancer. Two of these sites, denoted oligo-H and oligo-K, were shown by several criteria, including cell distribution and stimulation experiments, EMSA cross-competition studies, and proteolytic clipping bandshift assays, to bind to the same protein identical to the transcription factor B cell lineage-specific activator protein (BSAP) (NF-HB, S alpha-BP). To assess the possible functional role of these BSAP binding sites in the 3' alpha enhancer, we transiently transfected a construct containing a 314-bp 3' alpha enhancer fragment upstream of a luciferase reporter gene in MOPC-315 cells, a plasmacytoma line lacking BSAP. In these cells, co-transfection with a vector expressing recombinant BSAP led to significant reduction in the activity of the 3' alpha enhancer fragment. Conversely, in the mature B lymphoma cell line CH12.LX, a cell line that expresses BSAP and has a less active 3' alpha enhancer, selective BSAP down-regulation by an antisense phosphorothioate oligonucleotide was sufficient to considerably up-regulate 3' alpha enhancer activity, as were mutations of both binding sites that prevented binding of BSAP to the 3' alpha enhancer. Our findings thus suggest that the natural loss of BSAP expression in terminally differentiated plasma cells contributes to the activation of the murine Ig 3' alpha enhancer.

Animals↗

[Staging of rectal cancer: assessment with Gd-DTPA enhanced MR imaging].

This study was designed to assess the value of Gd-DTPA enhanced imaging in the staging of rectal carcinoma. Twenty-five patients were prospectively evaluated with MR imaging using a 1.5 Tesla unit. Based on the results of a barium study and/or digital examination, a balloon catheter was inserted to the level of the lesion before examination. Both T1- and T2-weighted images and Gd-DTPA enhanced images were obtained in all patients, and Gd-DTPA enhanced fat-saturation images were obtained in 11 patients. When both T1- and T2-weighted images were used, 24% of tumors were ill defined. However, when Gd-DTPA and Gd-DTPA enhanced fat-saturation imaging was added, all tumors were well defined and the quality of the images increased. There was no significant difference between images in the detection of muscular invasion. In the detection of perirectal fat invasion, however Gd-DTPA fat-saturation imaging showed increased sensitivity. Nevertheless, it also decreased specificity, so there was no significant difference between images. In the detection of metastatic lymph nodes, Gd-DTPA enhanced and Gd-DTPA enhanced fat-saturation imaging were not useful. Tumor detection using Gd-DTPA enhanced fat-saturation images was excellent. However, the accuracy of staging was not improved by supplemental Gd-DTPA enhancement.

Adult↗

[Late phase contrast enhanced-CT analysis of thrombosed type aortic dissection].

Seven patients with thrombosed type aortic dissection who underwent both early- and late-phase contrast enhanced (CE)-CT scans were analyzed. The image acquisition of early-phase CE-CT began 30 seconds after the intravenous administration of contrast material at an injection rate of 1.5 ml per second. Late-phase CE-CT began 6 minutes after contrast material injection. The thrombosed false lumens were not enhanced on early-phase CE-CTs in any of the cases. In five of seven cases, on the other hand, false lumens of the descending aorta were enhanced on late-phase images. Late enhancement in the false lumen was roughly divided into two patterns; (1) a crescentic enhancement of the sub-adventitial region (3 cases), and (2) a vague enhancement around the ulcerlike projection (2 cases). The mechanism of late enhancement in the false lumen was not fully elucidated. Although the clinical significance of late enhancement has not yet been established, it will be useful to demonstrate an unstable status of the thrombosed false lumens before organization.

Aged↗

Regulation of 3' IgH enhancers by a common set of factors, including kappa B-binding proteins.

Targeted disruption of the p50 subunit of NF-kappa B resulted in isotype class switch defects resembling those observed in mice in which the downstream IgH enhancer 3'alpha E(hs1,2) was deleted. We postulated that kappa B binding proteins may regulate class switching by interacting with 3'alpha E(hs1,2) or with other IgH 3' enhancers with which 3'alpha E(hs1,2) synergizes. kappa B binding sites were identified in 3'alpha E(hs1,2) and 3' alpha-hs4, the distal 3' IgH enhancer. A kappa B binding site within 3'alpha E(hs1,2) contributes to at least half the activity of the enhancer in plasma cells, while the same kappa B binding site participates in the complex repression of the enhancer in B cells. In the case of 3'alpha-hs4, a kappa B binding complex activates the enhancer in pre-B, B cells and plasma cells. Additional binding sites within 3'alpha-hs4 for factors known to regulate 3'alpha E(hs1,2), including Oct-1 and BSAP, were identified, and their contribution to 3'alpha-hs4 regulation during B cell development was assessed. Oct-1 positively regulates the enhancer in pre-B and B cells, while BSAP is a repressor in pre-B cells and an activator at the B cell stage. These studies identify kappa B binding proteins as key modulators of 3'alpha E(hs1,2) and 3'alpha-hs4, and suggest coregulation of the two enhancers by a common set of factors.

Animals↗

Structural and functional conservation of the Drosophila doublesex splicing enhancer repeat elements.

We have compared the RNA sequences and secondary structures of the Drosophila melanogaster and Drosophila virilis doublesex (dsx) splicing enhancers. The sequences of the two splicing enhancers are highly divergent except for the presence of nearly identical 13-nt repeat elements (six in D. melanogaster and four in D. virilis) and a stretch of nucleotides at the 5' and 3' ends of the enhancers. In vitro RNA structure probing of the two enhancers revealed that the 13-nt repeats are predominantly single-stranded. Thus, both the primary sequences and single-stranded nature of the repeats are conserved between the two species. The significance of the primary sequence conservation was demonstrated by showing that the two enhancers are functionally interchangeable in Tra-/Tra2-dependent in vitro splicing. In addition, inhibition of splicing enhancer activity by antisense oligonucleotides complementary to the repeats demonstrated the importance of the conserved single-stranded structure of the repeats. In vitro binding studies revealed that Tra2 interacts with each of the D. melanogaster repeat elements, except for repeat 2, with affinities that are indistinguishable, whereas Tra binds nonspecifically to the enhancer. Taken together, these observations indicate that the organization of sequences within the dsx splicing enhancers of D. melanogaster and D. virilis results in a structure in which each of the repeat elements is single-stranded and therefore accessible for specific recognition by the RNA-binding domain of Tra2.

Alternative Splicing↗

Bent DNA in the human adenovirus type 2 E1A enhancer is an architectural element for transcription stimulation.

The upstream half of the human adenovirus type 2 enhancer adopts a curved DNA structure. Most of the enhancer elements are within the curvature, suggesting that this unusual structure is linked to enhancer function. To verify this experimentally, I constructed in vitro transcription assay systems which could distinguish any effects generated by conformational changes in a DNA template. The curved DNA conformation in the enhancer clearly affected the extent of the stimulation of the E1A gene transcription: assays using the wild-type DNA template showed that the moderately curved enhancer was superior to the highly curved enhancer in transcriptional stimulation. In additional experiments, the enhancer region was substituted with a curved DNA derived from the bacteriophage lambda origin of replication. Assays using this mutant revealed that this curved segment could also act as an enhancer when it had the proper conformation. Consequently, DNA conformation may play a general role in transcriptional stimulation.

Adenovirus E1A Proteins↗

TATA box and Sp1 sites mediate the activation of c-myc promoter P1 by immunoglobulin kappa enhancers.

In Burkitt's lymphoma (BL) cells the proto-oncogene c-myc is transcriptionally activated by chromosomal translocation to the immunoglobulin (Ig) gene loci. This activation is characterized by preferential transcription from the c-myc promoter P1 and accomplished by juxtaposed Ig enhancer elements. To identify promoter elements required for enhancer-activated P1 transcription, we studied the activation of c-myc reporter gene constructs by the Ig kappa intron and 3' enhancers. Deletion analysis defined the core promoter with a TATA box and two adjacent GC/GT boxes upstream sufficient for basal and enhancer-activated transcription. Gel retardation assays revealed Sp1's binding affinity to the GC/GT box proximal to the TATA box to be higher than to the distal one. This difference correlated well with the resulting levels of transcription mediated by Sp1 in contransfection experiments in BL and Sp1-deficient SL2 cells. Sp3 also bound to the core promoter in vitro, but failed to transactivate in vivo. Mutation of the distal Sp1 site moderately affected basal transcription concomitant with a modest decrease in enhancer stimulation. Mutation of the proximal Sp1 site almost entirely abolished basal as well as enhanced transcription. A considerable level of basal transcription was maintained upon mutation of the TATA box, whereas enhancer-activated transcription largely was abolished. Stable transfection of the BL cell line Raji with constructs containing core promoter mutations confirmed that the proximal Sp1 site and the TATA box are essential for the activation of promoter P1 by the Ig kappa enhancers.

Burkitt Lymphoma↗

Three-dimensional FISP imaging in the evaluation of carotid cavernous fistula: comparison with contrast-enhanced CT and spin-echo MR.

PURPOSE: The purpose of this study was to assess the value of three-dimensional fast imaging with steady-state precession (FISP) MR sequences relative to contrast-enhanced CT and spin-echo MR imaging in the diagnosis of carotid cavernous fistula (CCF). METHODS: Seventeen patients with 19 angiographically proved CCFs had contrast-enhanced CT, spin-echo MR imaging, and 3-D FISP imaging. Three observers assessed these imaging studies as well as those of 43 control sides in a blinded manner for the presence or absence of CCF. Receiver operating characteristic analysis was used to assess the diagnostic utility of each imaging technique. In a nonblinded study, contrast-enhanced 3-D FISP images were also evaluated. RESULTS: Higher diagnostic accuracy was obtained with 3-D FISP sequences, as the shunt flow within the cavernous sinus was well seen. Sensitivity of 3-D FISP images was 83% and specificity was 100% in the blinded study. In the receiver operating characteristic analysis, the diagnostic performance of observers was found to be better with the 3-D FISP images than with the spin-echo MR images. Although there were no significant difference between 3-D FISP and contrast-enhanced CT, higher diagnostic performance was obtained with 3-D FISP images. In three CCFs without anterior drainage, a diagnosis was made only from the 3-D FISP images. The contrast-enhanced 3-D FISP images were not helpful, since the cavernous sinuses enhanced. CONCLUSION: Three-dimensional FISP imaging is superior to spin-echo MR imaging and contrast-enhanced CT in the diagnosis of CCF. Contrast-enhanced 3-D FISP images are not helpful for the evaluation of CCF.

Adolescent↗