ENTPD5: identification of splicing variants and their impact on cancer survival.

NTPDase5 is a nucleotidase of the endoplasmic reticulum that plays an important role in proteostasis as a regulator of protein N-glycosylation. This enzyme was first identified in hamster as a proto-oncogene activated upon a single nucleotide deletion that causes a frameshift leading to a truncated protein. Truncated NTPDase5 proteins were detected in human samples, but an oncogene was never identified. Searching for transcript variants in the GenBank database and using TCGA data, we discovered that splice variants could originate truncated human NTPDase5 proteins. We identified three main splicing events in the ENTPD5 gene: alternative acceptors, exon skipping, and alternative terminators. The analysis of impact of splicing events in cancers showed that skipping of exon 11-the event that leads to truncated proteins similar in size to the hamster oncogene-does not affect the hazard ratio of most tumors and was, in fact, a protective factor in the only two cancer studies where it was significant. We also identified four main patterns of impact of ENTPD5 in cancer and a potential variant-specific regulation by miR-215. Our findings shed light on a two-decade uncertainty about the origin of truncated NTPDase5 and contribute to the characterization of its impacts in cancer.

Cervical cancer stem-like cells: systematic review and identification of reference genes for gene expression.

Cervical cancer is the fourth most common cancer affecting women worldwide. Among many factors, the presence of cancer stem cells, a subpopulation of cells inside the tumor, has been associated with a worse prognosis. Considering the importance of gene expression studies to understand the biology of cervical cancer stem cells (CCSC), this work identifies stable reference genes for cervical cancer cell lines SiHa, HeLa, and ME180 as well as their respective cancer stem-like cells. A literature review was performed to identify validated reference genes currently used to normalize RT-qPCR data in cervical cancer cell lines. Then, cell lines were cultured in regular monolayer or in a condition that favors tumor sphere formation. RT-qPCR was performed using five reference genes: ACTB, B2M, GAPDH, HPRT1, and TBP. Stability was assessed to validate the selected genes as suitable reference genes. The evaluation validated B2M, GAPDH, HPRT1, and TBP in these experimental conditions. Among them, GAPDH and TBP presented the lowest variability according to the analysis by Normfinder, Bestkeeper, and ΔCq methods, being therefore the most adequate genes to normalize the combination of all samples. These results suggest that B2M, GAPDH, HPRT1, and TBP are suitable reference genes to normalize RT-qPCR data of established cervical cancer cell lines SiHa, HeLa, and ME180 as well as their derived cancer stem-like cells. Indeed, GAPDH and TBP seem to be the most convenient choices for studying gene expression in these cells in monolayers or spheres.

Extracellular Nucleotide Hydrolysis in Dermal and Limbal Mesenchymal Stem Cells: A Source of Adenosine Production.

Human Limbal (L-MSCs) and Dermal Mesenchymal Stem Cell (D-MSCs) possess many properties that increase their therapeutic potential in ophthalmology and dermatology. It is known that purinergic signaling plays a role in many aspects of mesenchymal stem cells physiology. They release and respond to purinergic ligands, altering proliferation, migration, differentiation, and apoptosis. Therefore, more information on these processes would be crucial for establishing future clinical applications using their differentiation potential, but without undesirable side effects. This study evaluated and compared the expression of ecto-nucleotidases, the enzymatic activity of degradation of extracellular nucleotides and the metabolism of extracellular ATP in D-MSCs and L-MSCs, isolated from discard tissues of human skin and sclerocorneal rims. The D-MSCs and L-MSCs showed a differentiation potential into osteogenic, adipogenic, and chondrogenic lineages and the expression of markers CD105+ , CD44+ , CD14- , CD34- , CD45- , as expected. Both cells hydrolyzed low levels of extracellular ATP and high levels of AMP, leading to adenosine accumulation that can regulate inflammation and tissue repair. These cells expressed mRNA for ENTPD1, 2, 3, 5 and 6, and CD73 that corresponded to the observed enzymatic activities. Thus, considering the degradation of ATP and adenosine production, limbal MSCs are very similar to dermal MSCs, indicating that from the aspect of extracellular nucleotide metabolism L-MSCs are very similar to the characterized D-MSCs. J. Cell. Biochem. 118: 2430-2442, 2017. © 2017 Wiley Periodicals, Inc.

Combined chemical shift imaging with early dynamic serial gadolinium-enhanced MRI in the characterization of adrenal lesions.

OBJECTIVE: The purpose of this study was to retrospectively evaluate early dynamic serial gadolinium-enhanced and chemical-shift imaging (CSI) MRI to distinguish benign from malignant adrenal tumors. MATERIALS AND METHODS: Between July 2007 and December 2011, 205 patients with 239 adrenal lesions (177 adenomas, 54 metastases, 5 pheochromocytomas, and 3 adrenal cortical carcinomas) underwent early dynamic serial gadolinium-enhanced MRI and CSI. CSI was assessed qualitatively and by calculating the adrenal index, and enhancement patterns were evaluated qualitatively and quantitatively. Statistical analyses were performed. RESULTS: Most adenomas exhibited either an arterial blush or homogeneous enhancement, whereas most metastases showed early peripheral or heterogeneous enhancement. Visualization of higher enhancement on arterial and venous phases enabled differentiation of adenomas from metastases in most cases. Moderate to high signal intensity drop on CSI was seen in 95.4% of adenomas and 14.8% of metastases. In lesions with this level of signal intensity drop, 87 of 88 lesions with a capillary blush were adenomas. Early dynamic serial imaging alone was a significant (p < 0.0001) indicator of nonadenoma (area under the curve [AUC], 0.912) with optimal sensitivity of 81% and specificity of 93% for differentiating adenomas from nonadenomas. Combined analysis (CSI and early dynamic serial imaging) was also significant (p < 0.0001 and p=0.0014, respectively) for diagnosing nonadenomas (AUC, 0.983) with optimal sensitivity of 94% and specificity of 98%. CONCLUSION: Early dynamic serial gadolinium-enhanced MRI aids in characterization of adrenal tumors, especially lesions that are categorized as indeterminate on the basis of CSI.

Post-contrast T1-weighted sequences in pediatric abdominal imaging: comparative analysis of three different sequences and imaging approach.

BACKGROUND: Post-contrast T1-weighted imaging is an essential component of a comprehensive pediatric abdominopelvic MR examination. However, consistent good image quality is challenging, as respiratory motion in sedated children can substantially degrade the image quality. OBJECTIVE: To compare the image quality of three different post-contrast T1-weighted imaging techniques-standard three-dimensional gradient-echo (3-D-GRE), magnetization-prepared gradient-recall echo (MP-GRE) and 3-D-GRE with radial data sampling (radial 3-D-GRE)-acquired in pediatric patients younger than 5 years of age. MATERIALS AND METHODS: Sixty consecutive exams performed in 51 patients (23 females, 28 males; mean age 2.5 ± 1.4 years) constituted the final study population. Thirty-nine scans were performed at 3 T and 21 scans were performed at 1.5 T. Two different reviewers independently and blindly qualitatively evaluated all sequences to determine image quality and extent of artifacts. RESULTS: MP-GRE and radial 3-D-GRE sequences had the least respiratory motion (P < 0.0001). Standard 3-D-GRE sequences displayed the lowest average score ratings in hepatic and pancreatic edge definition, hepatic vessel clarity and overall image quality. Radial 3-D-GRE sequences showed the highest scores ratings in overall image quality. CONCLUSIONS: Our preliminary results support the preference of fat-suppressed radial 3-D-GRE as the best post-contrast T1-weighted imaging approach for patients under the age of 5 years, when dynamic imaging is not essential.

Enhancement of the liver and pancreas in the hepatic arterial dominant phase: comparison of hepatocyte-specific MRI contrast agents, gadoxetic acid and gadobenate dimeglumine, on 3 and 1.5 Tesla MRI in the same patient.

PURPOSE: To evaluate the relative enhancement of liver, pancreas, focal nodular hyperplasia (FNH), pancreas-to-liver index, and FNH-to-liver index in the hepatic arterial dominant phase (HADP) after injection of hepatocyte-specific MRI contrast agents, gadoxetic acid and gadobenate dimeglumine, on 3 and 1.5 Tesla (T) MRI in the same patient. MATERIALS AND METHODS: The MRI database was retrospectively searched to identify consecutive patients who underwent abdominal MRI at 3T and 1.5T systems, using both 0.025 mmol/kg gadoxetic acid-enhanced and 0.05 mmol/kg gadobenate dimeglumine-enhanced MRI at the same magnetic strength field system. 22 patients were identified, 10 were scanned at 3T system and 12 at 1.5T system. The enhancement of liver, pancreas, and FNH was evaluated quantitatively on MR images. RESULTS: The relative enhancement of liver in HADP in the gadobenate dimeglumine-enhanced group in all subjects was significantly higher than that in gadoxetic acid-enhanced group (P = 0.023). The gadobenate dimeglumine-enhanced group in HADP had better relative enhancement of pancreas and FNH, pancreas-to-liver index, and FNH-to-liver index than gadoxetic acid-enhanced group, but the difference was not statistically significant. CONCLUSION: The 0.05 mmol/kg gadobenate dimeglumine-enhanced abdominal MRI studies at 3T and 1.5T MR systems are superior in relative enhancement of the liver in HADP to 0.025 mmol/kg gadoxetic acid-enhanced MRI. This type of assessment may provide comparative effectiveness data.

Liver-vessel cancellation artifact on in-phase and out-of-phase MRI imaging: a sign of ultra-high liver fat content.

PURPOSE: To describe a new MRI sign, the liver-vessel cancellation artifact, on In-Phase and Out-of-Phase gradient-echo sequences related to ultra-high liver fat content (>90%) by qualitative histology. MATERIALS AND METHODS: Institutional review board approval was obtained for this retrospective HIPAA-compliant study with waived informed consent. Patients with liver steatosis were searched in MRI (n = 195) and pathology (n = 116) databases between January 1, 2008, and June 20, 2010. Two readers blindly reviewed all MR images for the presence of the liver-vessel cancellation sign. Cross-reference of patients with biopsy-proven steatosis and MRI within one month was performed (n = 54; 25 males, 29 females; mean age 41.0 ± 18.9), with a population of 6 patients with ultra-high liver fat content (1 male, 5 females; mean age 15.5 ± 11.2). Performance diagnostic tests, including sensitivity and specificity, were performed. RESULTS: Liver-vessel cancellation sign was present in all patients with ultra-high liver fat content but in none of the remaining patients. Calculated sensitivity and specificity for the detection of ultra-high liver fat content with this sign were 100% (95% confidence interval [CI]: 69.1-100%) and 100% (95% CI: 98.4-100%), respectively. CONCLUSION: The presence of liver-vessel cancellation artifact around intra-hepatic vessels is a feature of ultra-high liver fat content.

Comparison of T1-weighted in- and out-of-phase single shot magnetization-prepared gradient-recalled-echo with three-dimensional gradient-recalled-echo at 3.0 Tesla: preliminary observations in abdominal studies.

PURPOSE: To describe in-phase (IP)/out-of-phase (OP) imaging with single shot magnetization-prepared gradient-recalled-echo (MP-GRE) and to compare intra-individually IP/OP MP-GRE with IP/OP three-dimensional gradient-recalled-echo (3D-GRE) at 3.0 Tesla (T). MATERIALS AND METHODS: Thirty-six subjects (15 males, 21 females; mean age 46.97 ± 14.97) who had abdominal MRI examinations including precontrast T1-weighted IP/OP MP-GRE, IP/OP 3D-GRE were included in the study. Two radiologists independently evaluated the sequences qualitatively for extent of artifacts, lesion detectability, and conspicuity and subjective grading of liver steatosis. Quantitative evaluation was performed by one radiologist and included liver fat index, liver and spleen SNR, and liver-lesion and liver-spleen CNR. RESULTS: Respiratory ghosting was more pronounced on 3D-GRE (P < 0.0008). The degrees of parallel imaging residual artifacts, shading and blurring were significantly higher on the 3D-GRE sequences (P < 0.0008). Spatial misregistration and bounce point artifacts were only observed with MP-GRE images. Pixel graininess was more apparent on MP-GRE (P < 0.0008). Lesion detectability, confidence, and conspicuity were considerably higher on MP-GRE. Visual appreciation of steatosis was superior on 3D-GRE. Overall image quality was superior on MP-GRE (P < 0.0008). CONCLUSION: Higher image quality and improved lesion detectability were present with IP/OP MP-GRE technique. Inversion-recovery prepared techniques may represent an important evolution for precontrast T1-weighted image at 3.0T. The good image quality of MP-GRE sequences acquired in a free breathing manner should recommend its use in patients unable to suspend breathing.

Combined hepatocellular carcinoma-cholangiocarcinoma: report of MR appearance in eleven patients.

PURPOSE: To describe the magnetic resonance imaging (MRI) appearance of hepatic combined hepatocellular-cholangiocarcinoma (cHCC-CC) in 11 patients. MATERIALS AND METHODS: The database of the Department of Pathology was cross-referenced with the MRI database to identify patients with confirmed cHCC-CC who had undergone MRI. Eleven consecutive patients were included (seven female, mean age 57.6 years). All patients were Caucasian. Five of 11 had a clinical history of chronic liver disease. Two radiologists retrospectively analyzed the images in consensus. RESULTS: cHCC-CC was a single mass in 10/11 patients. Margins were well-defined in 6/11. All tumors showed heterogeneous hyperintensity on T2. On postcontrast imaging, 6/11 showed early ring-enhancement (four noncirrhotic), with progressive enhancement in central portions, and 5/11 patients showed a diffuse heterogeneous early enhancement. Three of these five were cirrhotic and displayed partial washout with portions of contrast retention on later phases. Other findings included: late capsule enhancement (two patients), biliary dilatation (one), venous invasion (three), lymphadenopathy (six), and findings of cirrhosis (five). CONCLUSION: cHCC-CC presents most commonly as a single mass. Five of 11 had MR findings of cirrhosis. Clues to the diagnosis were moderately high signal on T2, portions of tumor that show progressive enhancement/contrast retention, and frequent lack of capsule. Early enhancement patterns included early ring-enhancement and diffuse heterogeneous enhancement.

MR findings of lithium-related kidney disease: preliminary observations in four patients.

PURPOSE: To describe MR features of the kidney in patients on chronic lithium therapy and to correlate findings with the level of renal impairment. METHOD: In this retrospective HIPPA compliant study, a search was performed in our institutional clinical and radiological computerized database for subjects with lithium-related kidney disease between August 1, 2009 and May 30, 2010. Four patients (2 male and 2 females, mean age 64.75 ± 3.5) with a total of eight kidneys fulfilled the search criteria. T2-weighted images were used to evaluate the presence, size, and distribution of renal cysts. T1-weighted images were used to evaluate kidney length, parenchymal thickness, and cortico-medullary differentiation (CMD). RESULTS: All kidneys displayed multiple, very abundant, small size (1-2 mm) cysts. The distribution of the cysts was symmetric in renal cortex and medulla and the number of cysts was similar in both kidneys. Sparse, asymmetrical parenchymal renal cysts >3 mm in diameter were also observed. The size and parenchyma thickness of both kidneys was considered normal in all patients. The CMD differentiation was preserved only in patients with normal laboratory kidney findings (n = 2), but was lost in patients with chronic renal failure. CONCLUSIONS: Multi-microcystic kidney disease secondary to long-term lithium therapy can be detected with MR imaging regardless of known renal impairment. Preservation of renal CMD was observed in both patients with normal kidney function. The results of our preliminary study suggest the possible role of MR imaging for the screening of early manifestations of nephropathy in patients undergoing chronic lithium therapy.

In-phase and out-of-phase gradient-echo imaging in abdominal studies: intra-individual comparison of three different techniques.

BACKGROUND: T1-weighted gradient-echo in-phase and out-of-phase imaging is an essential component of comprehensive abdominal MR exams. It is useful for the study of fat-containing lesions and to identify various disease states related to the presence of fat in the liver. PURPOSE: To compare three T1-weighted in-phase and out-of-phase (IP/OP) gradient-echo imaging sequences in an intra-individual fashion, and to determine whether advantages exist for each of these sequences for various patient types. MATERIAL AND METHODS: One hundred and eighteen consecutive subjects (74 men, 44 women; mean age 53.9 ± 13.8 years) who had MRI examinations containing all three different IP/OP sequences (two-dimensional spoiled gradient-echo [2D-GRE], three-dimensional gradient-echo [3D-GRE], and magnetization-prepared gradient-recall echo [MP-GRE]) were included. Two different reviewers independently and blindly qualitatively evaluated IP/OP sequences to determine image quality, extent of artifacts, lesion detectability and conspicuity, and subjective grading of liver steatosis for the various sequences. Quantitative analysis was also performed. Qualitative and quantitative data were subjected to statistical analysis. RESULTS: Respiratory ghosting, parallel imaging, and truncation artifacts as well as shading and blurring were more pronounced with 3D-GRE IP/OP imaging. Overall image quality was higher with 2D-GRE (P < 0.05). Detectability of low-fluid content lesions was lower with IP/OP MP-GRE sequences. MP-GRE sequences had the lowest SNRs (P < 0.001). Liver-to-spleen and liver-to-lesion CNRs were significantly lower with 3D-GRE and MP-GR, respectively (P < 0.001). Fat liver indexes showed strongly positive correlation between all sequences. CONCLUSION: Currently, 2D-GRE remains the best approach for clinical IP/OP imaging. The good image quality of MP-GRE sequences acquired in a free-breathing manner should recommend its use in patients unable to suspend breathing.

Lysosomal enzyme trafficking factor LYSET enables nutritional usage of extracellular proteins.

Mammalian cells can generate amino acids through macropinocytosis and lysosomal breakdown of extracellular proteins, which is exploited by cancer cells to grow in nutrient-poor tumors. Through genetic screens in defined nutrient conditions, we characterized LYSET, a transmembrane protein (TMEM251) selectively required when cells consume extracellular proteins. LYSET was found to associate in the Golgi with GlcNAc-1-phosphotransferase, which targets catabolic enzymes to lysosomes through mannose-6-phosphate modification. Without LYSET, GlcNAc-1-phosphotransferase was unstable because of a hydrophilic transmembrane domain. Consequently, LYSET-deficient cells were depleted of lysosomal enzymes and impaired in turnover of macropinocytic and autophagic cargoes. Thus, LYSET represents a core component of the lysosomal enzyme trafficking pathway, underlies the pathomechanism for hereditary lysosomal storage disorders, and may represent a target to suppress metabolic adaptations in cancer.

Hepatic tumor response evaluation by MRI.

Noninvasive evaluation of hepatic tumor response is necessary to improve the survival rate and quality of life of cancer patients. Among radiologic imaging modalities, MRI plays a significant role in the management of patients with hepatic tumor and is crucial for diagnosis, treatment planning and assessment of response or recurrence, because of its high contrast resolution, lack of ionizing radiation and the possibility of performing functional imaging sequences. This review provides an overview of the MRI findings after various treatments in patients with primary and secondary focal liver malignancies. The imaging methods described focus on the recent trends of using MRI techniques as biomarkers for disease. We also describe the appearance of successful and incomplete response for the various forms of treatment, including transcatheter arterial chemoembolization, ablative therapy, systemic chemotherapy and radiation therapy. Dynamic contrast-enhanced MRI is regarded as an established noninvasive method and potential biomarker for tumor detection, as well as for the characterization of the tumor response. Diffusion-weighted MRI, perfusion-weighted MRI and MRS are also promising functional biomarkers to help select patients for various therapies and to assess the response to treatments. However, further validation and standardization should be performed before their widespread use as imaging biomarkers.

Comparison of a single shot T1-weighted in- and out-of-phase magnetization prepared gradient recalled echo with a standard two-dimensional gradient recalled echo: preliminary findings.

PURPOSE: To compare in-phase (IP)/out-of-phase (OP) single shot magnetization-prepared gradient-recalled-echo (MP-GRE) with a standard two-dimensional gradient-recalled-echo (2D-GRE), and to compare image quality of MP-GRE in cooperative and noncooperative subjects. MATERIALS AND METHODS: Ninety-six consecutive subjects (52 males, 44 females; mean age, 53.2 ± 16.7 years), both cooperative (n = 73) and noncooperative (n = 23) subjects who had MRI examinations including precontrast T1-weighted IP/OP MP-GRE with or without IP/OP 2D-GRE were included in the study. The sequences were independently qualitatively evaluated by two radiologists. Quantitative analysis of liver fat index, signal-to-noise ratio (SNR) and liver-lesion contrast-to-noise ratio (CNR) was also performed. Data were subjected to statistical analysis. RESULTS: The visual detection of the presence or absence of liver steatosis showed no differences between 2D-GRE and MP-GRE imaging (k = 1). Minor differences were observed on image quality between MP-GRE and 2D-GRE in cooperative subjects, and between MP-GRE sequences performed in cooperative and noncooperative subjects. Liver fat index results were strongly positively correlated (r = .98; 95% confidence interval [CI] 0.97 to 0.98; P < .0001). Intercept (.14; 95% CI .13 to .15; P < .0001) and slope (.83; 95% CI .79 to .86; P < .0001) were statistically significant. CONCLUSION: IP/OP MP-GRE and 2D-GRE comparably demonstrate the presence or absence of hepatic steatosis. Image quality of MP-GRE was also comparable to 2D-GRE, and was not substantially adversely affected if subjects were unable to cooperate with breathholding instructions.

Bone metastases: evaluation of acuity of lesions using dynamic gadolinium-chelate enhancement, preliminary results.

PURPOSE: To evaluate whether enhancement on serial dynamic gadolinium-enhanced abdominal-pelvic MR imaging (DCE-MRI) can determine the acuity of bone metastases. MATERIALS AND METHODS: Twenty consecutive patients who underwent abdominal-pelvic DCE-MRI for evaluation/staging of a proven cancer and had bone metastases were included. Two radiologists analyzed in consensus 59 DCE-MRIs of these patients. Region of interest measurements were performed in up to three lesions on noncontrast T1-weighted, serial hepatic arterial dominant phase (HADP), early hepatic venous phase (EHVF), and interstitial phase (IP) postgadolinium images, and the percentage enhancement of 134 lesions was calculated. The coordinator separately and retrospectively sorted the lesions into three groups based on the imaging and clinical information: acute/active, subacute, and chronic metastases. RESULTS: The mean percentage enhancement of the bone metastases classified as acute/active, subacute, and chronic in the HADP, EHVP and IP were respectively (%): 134, 107, 99; 87, 86, 87; and 39, 65, 73. In the HADP, acute/active lesions enhanced significantly more than both subacute (1.53-fold) and chronic (3.4-fold) lesions (P < 0.01). Time intensity curves were significantly different between these three entities as well. CONCLUSION: The enhancement on arterial phase images and the time-intensity curves were different for acute/active, subacute, and chronic bone metastases.

Quarter-dose (0.025 mmol/kg) gadobenate dimeglumine for abdominal MRI in patients at risk for nephrogenic systemic fibrosis: preliminary observations.

OBJECTIVE: The purpose of this study is to evaluate the feasibility of 0.025 mmol/kg gadobenate dimeglumine, which is one quarter of the standard dose, for abdominal 3-T MRI studies in patients considered to be at risk for nephrogenic systemic fibrosis, using qualitative and quantitative measures and comparison with higher doses. MATERIALS AND METHODS: The MRI database was retrospectively searched to select consecutive patients who underwent quarter-dose gadobenate dimeglumine-enhanced abdominal MRI at 3 T, between January 1, 2009, and January 15, 2010, and who underwent half-dose (0.05 mmol/kg) gadobenate dimeglumine-enhanced abdominal MRI at 3 T during one randomly chosen month. There were 25 patients in the final quarter-dose group (16 men and nine women; mean age, 57 years) and 44 patients in the half-dose group (21 men and 23 women; mean age, 58 years). The enhancement of abdominal organs and aorta was evaluated qualitatively and quantitatively on contrast-enhanced images. The overall quality of abdominal enhancement was also evaluated. RESULTS: Reviewers rated the diagnostic enhancement of the evaluated organs in all phases of enhancement for both studied doses, but the half dose had significantly higher ratings than did the quarter dose in all comparisons (p, 0.034 to < 0.0001), except in the pancreas in the early hepatic venous phase (p = 0.095 for reviewer 1; p = 0.0611 for reviewer 2). The overall enhancement quality of the quarter dose was rated as good in all phases of enhancement, although it was significantly lower than that for the half dose (p ≤ 0.0001). The liver, pancreas, renal cortex, and aorta had 1.52-1.93-fold, 1.53-1.90-fold, 1.46-1.77-fold, and 1.58-1.84-fold, respectively, higher percentages of enhancement with the half dose than with the quarter dose (p, 0.0049 to < 0.0001). CONCLUSION: A one-quarter dose of gadobenate dimeglumine at 3 T is a feasible alternative for abdominal MRI in patients at risk for nephrogenic systemic fibrosis. Our results might have important clinical implications, because greater safety may be conferred on patients with poor renal function with this low dose of contrast agent.

Free-breathing 3D T1-weighted gradient-echo sequence with radial data sampling in abdominal MRI: preliminary observations.

OBJECTIVE: The purposes of this study were to evaluate the feasibility of a free-breathing 3D gradient-recalled echo sequence with radial data sampling (radial 3D GRE) in abdominal MRI compared with a standard 3D GRE volumetric interpolated breath-hold examination (VIBE) sequence for imaging of cooperative patients and to perform a preliminary assessment in imaging of noncooperative patients. MATERIALS AND METHODS: Fifty-five consecutively registered patients who underwent unenhanced and contrast-enhanced abdominal MRI with the free-breathing radial 3D GRE technique constituted the study population. Two readers independently and blindly evaluated the images. RESULTS: Overall image quality with the contrast-enhanced radial 3D GRE sequence was lower than but rated at least nearly as good as that with the 3D GRE VIBE sequence (p < 0.0001). Higher scores were recorded for 3D GRE VIBE images with respect to pixel graininess, streaking artifact, and sharpness (p = 0.0009 to p < 0.0001). Except for sharpness of vessels on unenhanced images, results for the radial 3D GRE sequence did not differ significantly in the comparison of cooperative and noncooperative patients (p = 0.004). For imaging of noncooperative patients, radial 3D GRE images of children had higher ratings for shading (unenhanced, p = 0.0004; contrast-enhanced, p < 0.0001) and streaking artifacts on contrast-enhanced images (p = 0.0017) than did those of adults. Overall image quality was higher for pediatric patients. In lesion analysis, use of the 3D GRE VIBE sequence was associated with significantly greater detectability, confidence, and conspicuity than was use of the radial 3D GRE sequence (p = 0.00026-0.011). CONCLUSION: A free-breathing radial 3D GRE sequence is feasible for abdominal MRI and may find application in imaging of patients who are unable to suspend respiration, especially children.

Characterization of adrenal lesions with 1.5-T MRI: preliminary observations on comparison of three in-phase and out-of-phase gradient-echo techniques.

OBJECTIVE: The purpose of this study was to use previously described quantitative evaluation methods to compare the performance of 3D gradient-recalled echo (GRE) and magnetization-prepared (MP) GRE in-phase and out-of phase sequences with standard 2D GRE technique in the characterization of adrenal lesions. MATERIALS AND METHODS: The study sample consisted of 44 consecutively registered patients (22 men, 22 women; mean age, 59.1 ± 11.6 years) with 50 adrenal lesions who underwent standard abdominal MRI that included in-phase and out-of-phase 2D GRE (n = 41), 3D GRE (n = 35), MP GRE (n = 36), or a combination of these techniques. The adrenal signal intensity (SI) index and adrenal-to-spleen, adrenal-to-liver, and adrenal-to-muscle SI ratios of each lesion were calculated and compared for the three techniques by independent samples Student t test. The area under the receiver operating characteristic (ROC) curve (AUC) for each evaluation method was determined, and comparisons of independent ROC curves were performed for all sequences. RESULTS: For all sequences, the mean adrenal SI index and SI ratios of adenomas and nonadenomas differed significantly (p < 0.001). For the 3D GRE and MP GRE techniques, adrenal SI index and modified adrenal-to-spleen ratio, respectively, had the larger AUCs, but the difference was not statistically significant. Different thresholds for the three techniques were recommended for discriminating adenoma from nonadenoma. CONCLUSION: The results of characterization of adrenal lesions with MP GRE and 3D GRE in-phase and out-of-phase MRI techniques are comparable to those obtained with the reference standard 2D GRE technique. Different thresholds should be selected according to the in-phase and out-of-phase techniques used and for the various evaluation methods.

Normalization in Human Glioma Tissue.

For tissues obtained from glioma samples with/without nonneoplastic brain there is no consensus for universal reference gene but there are some potential genes that might have good stability, under certain conditions. Considering all points described in this work, the care with tissue collection, until gene amplification, directly impacts on the reliable characterization of its mRNA levels. Moreover, it is clear the importance of selecting the most appropriate reference genes for each experimental situation, to allow the accurate normalization of target genes, especially for genes that are subtly regulated.