Regulatory role of kinases and phosphatases on the internalisation of caveolae in HepG2 cells

The caveolar cycle is thought to be regulated by synchronised function of kinases and phosphatases. Using ocadaic acid--a serine/threonine protein phosphatase inhibitor--and an inhibitor of tyrosine phosphatase (sodium orthovanadate) we have followed the internalisation of caveolae. Since albumin binding to its receptor (gp60) can induce pinching off of caveolae from the plasma membrane, we also used this physiological ligand to induce the internalisation. Our confocal microscopic results show that both ocadaic acid and vanadate treatments have significantly decreased caveolin (caveolin-1 and -2) labelling on the cell surface, while the cytoplasmic labelling became much stronger. Quite often large, strongly labelled "granules" appear at the perinuclear region. Very strong caveolin labelling was detected along the actin-cytoskeleton suggesting that caveolae might move along these filaments. Our electron microscopic results also show an intensive caveolae pinching off from the plasma membrane. After ocadaic acid and vanadate treatments the number of surface connected vesicles (caveolae) decreases. At the same time, large multivesicular bodies (termed caveosomes) appear in the perinuclear area of the cytoplasm. By immunoprecipitation and Western blot analysis we detect an increased tyrosine phosphorylation of a approximately 29kDa protein in ocadaic acid and vanadate treated samples. This protein was identified as caveolin-2. No significant change in the tyrosine phosphorylation of caveolin-1 was found. From these data we can conclude that caveolae internalisation is regulated by phosphorylation of caveolin-2.     

Recent advances in liver sinusoidal endothelial ultrastructure and fine structure immunocytochemistry

Ultrastructure reports have described that liver sinusoidal endothelial cell (LSEC)s contain a cytoskeletal framework of filamentous actin. Small G protein has emerged as an important regulator of the actin cytoskeleton, and consequently, of cell morphology and motility. We investigated actin filaments in relation to SEF in LSECs using a heavy meromyosin-decorated reaction and thereby elucidated the roles of small G protein and actin cytoskeleton in the morphological and functional alterations of SEF. Caveolin-1 expression has also been found in fenestrations with many characteristics of liver sinusoidal endothelial cells. Currently, fenestral studies and human disease are revealing ways to increase the liver sieve's porosity, which is reduced through pathological mechanisms. Hepatic sinusoidal endothelial dysfunction, which is known to impair endothelium-dependent relaxation in the liver microcirculation, contributes to increased intrahepatic vascular resistance.     

Quantitative MRI of the liver: Evaluation of extracellular volume fraction and other quantitative parameters in comparison to MR elastography for the assessment of hepatopathy

BackgroundChronic liver diseases pose a major health problem worldwide, while common tests for diagnosis and monitoring of diffuse hepatopathy have considerable limitations. Preliminary data on the quantification of hepatic extracellular volume fraction (ECV) with magnetic resonance imaging (MRI) for non-invasive assessment of liver fibrosis are encouraging, with ECV having the potential to overcome several of these constraints.PurposeTo clinically evaluate ECV provided by quantitative MRI for assessing the severity of liver disease.Materials and methodsIn this prospective study, multiparametric liver MRI, including T1 mapping and magnetic resonance elastography (MRE), was performed in subjects with and without hepatopathy between November 2018 and October 2019. T1, T2, T2*, proton density fat fraction and stiffness were extracted from parametric maps by regions of interest and ECV was calculated from T1 relaxometries. Serum markers of liver disease were obtained by clinical database research. For correlation analysis, Spearman rank correlation was used. ROC analysis of serum markers and quantitative MRI data for discrimination of liver cirrhosis was performed with MRE as reference standard.Results109 participants were enrolled (50.7 ± 16.1 years, 61 men). ECV, T1 and MRE correlated significantly with almost all serum markers of liver disease, with ECV showing the strongest associations (up to r = 0.67 with MELD, p < 0.01). ECV and T1 correlated with MRE (0.75 and 0.73, p < 0.01 each). ECV (AUC 0.89, cutoff 32.2%, sensitivity 85%, specificity 87%) and T1 mapping (AUC 0.85, cutoff 592.5 ms, sensitivity 83%, specificity 75%) featured good performances in detection of liver cirrhosis with only ECV performing significantly superior to model of end stage liver disease (MELD), AST/ALT ratio and international normalized ratio (p < 0.01, respectively).ConclusionQuantification of hepatic extracellular volume fraction with MRI is suitable for estimating the severity of liver disease when using MRE as the standard of reference. It represents a promising tool for non-invasive assessment of liver fibrosis and cirrhosis.     

Caveolae and caveolin-1 in reptilian liver

Caveolae are plasma-membrane invaginations that, by interacting with membrane-associated molecules such as endothelial nitric oxide synthase and tyrosine kinases, precisely regulate cell-signalling pathways responsible for cell structure and cell function. Indeed, there is widespread evidence that caveolae associate, structurally and functionally, with proteins, lipids and solutes to facilitate transcellular transport of these macromolecules. Caveolin-1, one of the family of membrane proteins that form caveolae, is most prominently expressed in endothelial cells of the vascular bed. Therefore, we have applied advanced electron microscopy as well as molecular biology techniques to study the presence of caveolae and caveolin-1 in the liver sinusoidal endothelium of reptiles. Reptiles are known to store excess lipid in the liver as an energy source for hibernation, and so offer a useful animal model in which to assess the structural and functional implications these subcellular compartments might have on liver sinusoidal endothelial transport. This study demonstrates that caveolae are indeed conserved across vertebrate species, whether mammalian or reptilian. It also presents as first novel data on the presence of caveolin-1-associated, tubular structures located within the cytoplasm of the lizard liver sinusoidal endothelium.     

Ocadaic acid treatment causes tyrosine phosphorylation of caveolin-2 and induces internalization of caveolae in rat peritoneal macrophages

In this paper, we provide evidences that protein phosphatases could regulate the internalization cycle of caveolae in rat peritoneal cells. Ocadaic acid (OA)—a serine/threonine phosphatase inhibitor—was used in various concentrations (4 and 100 nM) to study the internalization of horseradish peroxidase (HRP) in resident and elicited macrophages. We have found that OA in both concentrations has significantly decreased HRP uptake in resident and elicited cells. The results of our morphometrical analysis showed that in OA-treated cells, the number of surface-connected caveolae has been dramatically decreased. Simultaneously large, endosome-like vacuoles containing small vesicles appeared in the cytoplasm. The membrane of these small vesicles was labeled with anti-caveolin-1 antibody. Immunoprecipitation and Western blot analysis revealed that in OA-treated cells a 29 kDa protein identified as caveolin-2 in macrophages was phosphorylated on tyrosine residues. These data support the idea that there is a close correlation between the phosphorylation of caveolin-2 and endocytosis of caveolae: the tyrosine phosphorylation of this 29 kDa protein can drive caveolae to pinch off from the plasma membrane and causes accumulation of caveolae in a multivesicular body-like cellular compartment, which was never found to contain lysosomal enzymes. As a result of OA treatment caveolin-2 remains phosphorylated and the phosphorylation of these protein might inhibit the recycling of caveolae.     

Superparamagnetic iron oxide (SPIO) enhancement in the cirrhotic liver: a comparison of two doses of ferumoxides in patients with advanced disease

The aim of this study was to establish whether enhancement of the liver by the MRI contrast agent ferumoxides could be effectively achieved at a reduced dose of 7.5 micromol/kg in patients with advanced liver cirrhosis. Forty-two liver transplant candidates with end-stage cirrhosis underwent SPIO-enhanced MRI at 1.5T, using either 15 micromol/kg or 7.5 micromol/kg ferumoxides. The lower dose of ferumoxides was also used in 21 non-cirrhotic patients with colorectal liver metastases who acted as a control group. The percentage signal intensity loss (PSIL) after SPIO was measured in all patients, and in those patients with tumors the post-SPIO contrast-to-noise ratio (CNR) was measured. The median PSIL after SPIO in the high dose cirrhotic (HDLC), low dose non-cirrhotic (LDNC) and low dose cirrhotic (LDLC) patients was 86.3%, 74.6%, and 64.2% respectively. These differences were significant using the Mann-Whitney U test. Tumors were found in 8 patients in the high dose cirrhotic group, 9 in the low dose cirrhotic group, and all 21 of the control group. No significant differences were found between the CNR values after SPIO in the 3 groups (median values HDLC 15.1, LDNC 23.7, LDLC 19.5). In patients with late-stage cirrhosis the PSIL after SPIO was significantly less at 7.5 micromol/kg than at 15 micromol/kg, but both doses produced a substantial loss of signal. Lesion to liver CNR was not adversely affected by using the lower dose, so when imaging at 1.5T the authors would recommend using 7.5 micromol/kg in patients with liver cirrhosis.     

13C-aminopyrine demethylation is decreased in cirrhotic patients with normal biochemical markers

This study determined the rates of ¹³C-aminopyrine metabolism in patients with varying degrees of liver cirrhosis as defined by clinical scores. Twenty-five cirrhotic patients and 18 healthy subjects underwent a ¹³C-aminopyrine breath test. The cumulative per cent dose recovery (cPDR) of ¹³C on breath expressed as a percentage of the administered dose at 2 h was significantly lower in cirrhotic patients than in healthy subjects (median: 1.7% versus 9.0%; p<.0001). Significant inverse associations between cPDR at 2 h and the model for end-stage liver disease score, Child–Pugh score, international normalised ratio and bilirubin (all p<.05), but not alanine aminotransferase or alkaline phosphatase were observed in the cirrhotic patients. Taking each biochemical marker independently, cirrhotic patients with normal biochemistry had a significantly lower cPDR at 2 h than healthy subjects (all p<.05). Differences in ¹³C-aminopyrine metabolism were evident in cirrhotic patients with less severe disease and may mark hepatic dysfunction when conventional biochemical markers appear unchanged.     

Hepatic sarcoidosis: MR appearances in patients with chronic liver disease

Purpose

To describe the MR appearances of hepatic sarcoidosis in patients with chronic liver disease and correlate the results with clinical stage of disease as measured with the Mayo end-stage liver disease (MELD) score.

Materials and methods

Twenty patients with chronic liver disease and histopathological diagnosis of hepatic sarcoidosis who underwent MR imaging were included in this study. Two abdominal radiologists retrospectively reviewed all images for the presence of cirrhosis, imaging pattern of the liver, intrahepatic biliary dilatation, presence of areas of parenchymal atrophy, presence of splenic nodules and lymphadenopathy. Imaging findings were correlated with the MELD score.

Results

Of the patients, 14/20 had imaging findings of cirrhosis, 9/20 had a large macronodular pattern of liver cirrhosis and 5/20 had a diffuse pattern of liver cirrhosis. Peripheral wedge-shaped areas of parenchymal atrophy were observed in 10 patients. The combination of a central macronodular pattern and peripheral atrophy was observed in 9/20 patients. The pattern of cirrhosis had statistically significant correlation with the presence of wedge-shaped areas of parenchymal atrophy (p < 0.005). No statistically significant difference was revealed between the clinical score of patients who had imaging findings consistent with cirrhosis and those who did not.

Conclusion

MR imaging appearances of chronic sarcoid liver disease are diverse and do not appear to correlate with severity of clinical disease. Large central regenerative nodules and wedge-shaped areas of peripheral parenchymal atrophy are frequent findings and may help to suggest the diagnosis.     

Caveolae and caveolin isoforms in rat peritoneal macrophages

Caveolea are special (highly hydrophobic) plasma membrane invaginations with a diameter of 50-100 nm. Their characteristic features are the flask- or omega-shape and the lack of basket-like coat composed of clathrin. Caveolin-an integral membrane protein-is the principal component of caveolae membranes in vivo. Multiple forms of caveolin have been identified: caveolin-1alpha, caveolin-1beta, caveolin-2 and caveolin-3. They differ in their specific properties and tissue distribution. In this paper we summarize the morphological and biochemical data providing strong evidence about the existence and function of caveolae in rat peritoneal macrophages. When studied electron microscopically, the surface of both resident and elicited macrophages exhibited omega- or flask-shaped plasma membrane invaginations. There was a significant difference, however, in the number of these profiles: whereas in resident cells only a small amount of them was found on the cell surface, in elicited cells they were abundantly present on the plasma membrane. Using an antibody against the VIP21/caveolin-1 isoform we showed that these plasma membrane pits were indeed caveolae. The number and the appearance of caveolae were found to be in close correlation with the functional activity of these phagocytotic cells, indicating that the formation of caveolae is a highly regulated process.Using Western blot analysis two different proteins ( approximately 29 and approximately 20 kDa)-both labelled with anti-caveolin antibodies-were identified in resident and elicited macrophages that have been isolated from rat peritoneal cavity. The approximately 20 kDa protein was labelled specifically only by anti-VIP21/caveolin-1, while the approximately 29 kDa protein was labelled by both anti-VIP21/caveolin-1 and anti-caveolin-2 antibodies. The presence of the approximately 29 kDa protein was highly characteristic of resident cells, and only a small amount of approximately 20 kDa protein was detected in these cells. Elicitation has resulted in a significant increase in the amount of approximately 20 kDa protein labeled only with anit-VIP21/caveolin-1. Our morphological (confocal and electron microscopical) studies have shown that in resident cells caveolin was present in the cytoplasm, in smaller vesicles and multivesicular bodies around the Golgi area. Only a very small amount of caveolae was found on the cell surface of these cells. In elicited macrophages, caveolae (labelled with anti-VIP21/caveolin-1 antibody) appeared in large numbers on the cell surface, but caveolin detected by anti-caveolin-2 was also found in small vesicles and multivesicular bodies. These data support the idea that the expression of the approximately 29 kDa (caveolin-related) protein is insufficient for caveolae formation in resident cells, it can function as a modified, macrophage-specific caveolin-2 isoform.Our results strongly suggest that caveolin-1 plays a crucial role in the formation of caveolae: it is the amount of caveolin-1 that regulates the appearance of caveolae on the plasma membrane.Studying the endocytotic processes of resident and elicited macrophages we have found that elicited macrophages bound and internalized significantly larger amounts of fluid phase marker (HRP) and immune complex (peroxidase-antiperoxidase-PAP) than resident cells. Serial section analysis, double labelled immunocytochemistry, and filipin treatment were used to demonstrate that caveolae can pinch off from the plasma membrane and can take part in endocytotic processes as alternative carriers in elicited macrophages.     

Effect of hepatic steatosis on native T1 mapping of 3T magnetic resonance imaging in the assessment of T1 values for patients with non-alcoholic fatty liver disease

PurposeThis study investigated whether T1 values in native T1 mapping of 3T magnetic resonance imaging (MRI) of the liver were affected by the fatty component.MethodsThis prospective study involved 340 participants from a population-based cohort study between May 8, 2018 and August 8, 2019. Data obtained included: (1) hepatic stiffness according to magnetic resonance elastography (MRE); (2) T1 value according to T1 mapping; (3) fat fraction and iron concentration from multi-echo Dixon; and (4) clinical indices of hepatic steatosis including body mass index, waist circumference, history of diabetes, aspartate aminotransferase, alanine aminotransferase, gamma-glutamyl transpeptidase, and triglycerides. The correlations between T1 value and fat fraction, and between T1 value and liver stiffness were assessed using Pearson's correlation coefficient. The independent two-sample t-test was used to evaluate the differences in T1 values according to the presence or absence of hepatic steatosis, and the one-way analysis of variance was used to evaluate the difference in T1 value by grading of hepatic steatosis according to MRI-based proton density fat fraction (PDFF). In addition, univariate and multivariate linear regression analyses were performed to determine whether other variables influenced the T1 value.ResultsT1 value showed a positive correlation with the fat fraction obtained from PDFF (r = 0.615, P < 0.001) and with the liver stiffness obtained from MRE (r = 0.370, P < 0.001). Regardless of the evaluation method, the T1 value was significantly increased in subjects with hepatic steatosis (P < 0.001). When comparing hepatic steatosis grades based on MRI-PDFF, the mean T1 values were significantly different in all grades, and the T1 value tended to increase as the grade increased (P < 0.001, P for trend <0.001). On multiple linear regression analysis, the T1 value was influenced by MRI-PDFF, calculated liver iron concentration, liver stiffness, and serum aspartate aminotransferase level.ConclusionThe T1 value obtained by current T1 mapping of 3T MRI was affected by the liver fat component and several other factors such as liver stiffness, iron concentration, and inflammation.     

Can preoperative diffusion-weighted MRI predict postoperative hepatic insufficiency after curative resection of HBV-related hepatocellular carcinoma? A pilot study

Liver fibrosis determines the functional liver reserve. Several studies have reported that the apparent diffusion coefficient (ADC) values of diffusion-weighted magnetic resonance imaging (DW-MRI) can assess liver fibrosis. We investigated whether DW-MRI predicts postoperative hepatic insufficiency and liver fibrosis in patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). Twenty-six patients with HBV-related HCC who received preoperative DW-MRI on a 3-T MRI system were enrolled between July and December 2008. ADC values were measured twice by two observers. Three “b values” were used: 50, 400 and 800 s/mm². Postoperative hepatic insufficiency was defined as persistent hyperbilirubinemia (total bilirubin level >5 mg/dl for more than 5 days after surgery) or postoperative death without other causes. The mean age (21 men and 5 women) was 51.4 years. Three patients experienced postoperative hepatic insufficiency. liver stiffness measurement predicted postoperative hepatic insufficiency, advanced fibrosis (F3–4), and cirrhosis significantly [area under the receiving operator characteristic curve (AUROC)=0.942, 0.771 and 0.818, respectively, with P=.047, 0.048 and 0.006, respectively]; ADC values of DW-MRI, however, did not (AUROC=0.797, 0.648 and 0.491, respectively, with P=.100, 0.313 and 0.938, respectively). Reliability of ADC values between right and left hepatic lobes (ρ=0.868 and ρ=0.910 in the first and second measures of Observer A; ρ=0.865 and ρ=0.831 in the first and second measures of Observer B) was high and the intra- and interobserver reliability (ρ=0.958 in observer A and ρ=0.977 in observer B; ρ=0.929 in the first measure and ρ=0.978 in the second measure between the two observers) were high. All reliability was significant (P<.001). Our results suggest that DW-MRI on a 3-T MRI system is not suitable for predicting postoperative hepatic insufficiency, advanced liver fibrosis, and cirrhosis in patients with HBV-related HCC, despite significantly high reliability.     

Hepatic nodules in liver transplantation candidates: MR imaging and underlying hepatic disease

OBJECTIVE: To assess by MR imaging the frequency of hepatic nodules in patients waiting on the liver transplant list and to determine whether certain underlying hepatic diseases were more often associated with the development of such hepatic nodules. MATERIAL AND METHODS: We reviewed the MR and clinical records in all patients seen by the liver transplant service at our center since its inception in January 1998 until September 2002. A total of 371 patients (207 men and 164 women, age range 18-68 years, mean 45 years) were included in the study. The presence of hepatic nodules, size, number and underlying hepatic diseases were determined in all patients. Magnetic resonance imaging was performed on a 1.5-T MR imager using T1-weighted, T2-weighted and multi-phase gadolinium-enhanced sequences. Odds ratio (OR) and 95% confidence intervals (CIs) were computed to evaluate the association between the underlying hepatic disease and the development of hepatic nodule. RESULTS: Among 371 liver transplantation candidates, the most common underlying hepatic disease was hepatitis C virus (HCV) infection, either alone (n=93; 25%) or associated with other hepatic diseases (n=40; 10.8%). Of all patients, 33 (8.9%) had regenerative nodules (RNs), 40 (10.7%) dysplastic nodules (DNs) and 57 (15.3%) hepatocellular carcinomas (HCCs). Hepatocellular carcinoma was observed in 35.3% of patients with HCV infection and alcohol abuse combined, 24.5% with cryptogenic cirrhosis, 25% with hemochromatosis and 19% with alcohol abuse. Patients who had either DNs or HCC were 2.5 times more likely to have either alcohol abuse or HCV, alone or combined, as the substrate of their liver disease (OR 2.54, 95% CI 1.56-4.13). Our data suggest a supra-additive interaction between HCV infection and ethanol in their association with MR imaging detected lesions. CONCLUSION: Patients with cryptogenic cirrhosis, alcohol abuse, HCV infection (alone or combined) and hemochromatosis had the greatest likelihood of having HCC, with the combination of HCV infection and alcohol abuse having the highest of all.     

Small hyperintense hepatic lesions on T1-weighted images in patients with cirrhosis: evaluation with serial MRI and imaging features for clinical benignity

PURPOSE: The aim of this study was to evaluate the frequency and magnetic resonance imaging (MRI) features of clinically benign, small (<2 cm) hyperintense hepatic lesions in the cirrhotic liver on T1-weighted MR images seen at serial MRI. MATERIALS AND METHODS: This study included 189 patients with cirrhosis, who underwent hepatic MRI more than twice with an interval of at least 12 months. The initial MR images were reviewed for the presence of small hyperintense lesions on T1-weighted images. The size, location and signal intensity on T2-weighted images as well as enhancement patterns of the corresponding lesions were recorded. RESULTS: On the initial T1-weighted MR images, 43 small hyperintense hepatic lesions were detected in 23 (12%) of 189 patients. Twelve (28%) of 43 lesions showed early enhancement and were pathologically diagnosed as hepatocellular carcinoma (HCC) during the follow-up period. Thirty-one (72%) of 43 lesions showed no early enhancement with various signal intensity on T2-weighted images (hyperintensity=4, isointensity=20, hypointensity=7). Among these 31 lesions, 12 showed no interval change, while 11 disappeared (n=10) or decreased in size (n=1). In the remaining eight lesions, seven were diagnosed as HCC on the basis of pathologic confirmation or the interval growth. CONCLUSION: Small hyperintense hepatic lesions on T1-weighted magnetic resonance (MR) images without early enhancement on the arterial-phase contrast-enhanced dynamic studies in patients with cirrhosis usually showed no interval growth or disappeared during the serial MRI. These lesions with additional findings of iso- or hypointensity on the T2-weighted MR images without "washout effect" on the contrast-enhanced equilibrium-phase images may more frequently be clinically benign or hyperplastic nodules than HCCs.     

Impact of diffusion-weighted MR imaging on the characterization of small hepatocellular carcinoma in the cirrhotic liver

Purpose

The purpose of this study was to determine whether or not adding diffusion-weighted magnetic resonance imaging (DWI) to conventional magnetic resonance (MR) imaging sequences improves the characterization of small hepatocellular carcinoma (HCC) (≤2 cm) in the setting of cirrhotic liver compared to conventional sequences alone.

Materials and Methods

A total of 62 cirrhotic liver patients with 82 nodules smaller than 2 cm in diameter were enrolled, and all lesions were pathologically confirmed. For the first reading session, which included precontrast T1- and T2-weighted images and T1 dynamic contrast-enhanced images, preindicated lesions by a study coordinator were characterized by two radiologists. They determined the confidence levels in consensus for the presence of small HCC into four grades. In another session, respiratory-triggered diffusion-weighted MR images (b factor=50, 400 and 800 s/mm²) were added to the previously reviewed images, and the same two radiologists again determined the confidence levels. The diagnostic performance of the combined DWI–conventional sequences set and the conventional sequences alone set was evaluated using receiver operating characteristic curves. Sensitivity and specificity values for characterizing small HCCs were also calculated.

Results

The area under the receiver operating characteristic curve for the second interpretation session (0.86) was significantly higher (P=.038) than that of the first session (0.76). The sensitivity was significantly increased from 75.7% to 87.8% by adding DWI to the conventional sequences (P=.015). No significant differences were observed for specificity values.

Conclusion

Adding DWI to conventional imaging modalities improves the diagnosis of small HCCs in the cirrhotic liver in terms of diagnostic performance and sensitivity by increasing reader confidence.     

基于新型支持向量机的影像组学在肝脏结节分类中的应用

肝癌是最常见的恶性肿瘤之一,亚洲地区最为常见的肝癌演变过程为肝炎-肝硬化结节-异型增生结节-肝细胞性肝癌．判断肝脏结节在演变过程所处分期,并采取干预措施,对降低肝癌的发生率非常关键．本文针对影像组学提出了更精确的支持向量机（SVM）分类算法——LFOA-F-SVM,用于对120名患者的腹部动态增强磁共振图像的肝脏结节进行四分类．该算法利用了考虑半径与几何间距的F-SVM,并结合莱维飞行策略（LF）的果蝇优化算法（FOA）寻求超参．为了验证方法的有效性,本文另外添加了5个UCI分类数据集（心脏、帕金森疾病、虹膜、葡萄酒和动物园）,并与SVM、PSO-SVM、FOA-SVM、F-SVM进行比较．结果表明,在6个分类数据集（包括肝脏结节数据集和5个UCI分类数据集）中,相对于其他分类算法,LFOA-F-SVM的分类准确率最高,在肝脏结节数据集中的四分类精确率和查全率也较高．     

R1ρ at high spin-lock frequency could be a complementary imaging biomarker for liver iron overload quantification

PurposeTo compare the correlations among the R1ρ, R2, and R2* relaxation rates with liver iron concentration (LIC) in the assessment of rat liver iron content and explore the application potential of R1ρ in assessing liver iron content.MethodsIron dextran (dosage of 0, 25, 50, 100, and 200 mg/kg body weight) was injected into 35 male rats to increase the amount of iron storage in the liver. After one week, all rats were euthanized with isoflurane. A portion of the largest hepatic lobe was extracted to quantify the LIC by inductively coupled plasma, and the remaining liver tissue was stored in 4% buffered paraformaldehyde for 24 h before MRI. Spin-lock preparation with a RARE (rapid acquisition with relaxation enhancement) readout (9 different spin-lock times and 7 different spin-lock frequencies (FSLs)) and multi-echo UTE (ultrashort TE) pulses were developed to quantify R1ρ and R2 * on a Bruker 11.7 T MR system. For comparisons with R1ρ and R2*, R2 was acquired using the CPMG sequence.ResultsMean R1ρ values displayed dispersion, with decrease in R1ρ at higher FSLs. Spearman's correlation analysis (two-tailed) indicated that the R1ρ values were significantly associated with LIC at FSL = 2000, 2500, and 3000 Hz (r = 0.365 and P = 0.031, r = 0.608 and P < 0.001, and r = 0.764 and P < 0.001, respectively), and were not significantly associated with LIC at FSL = 500, 1000, 1250, and 1500 Hz (all P > 0.05). R2 and R2* showed significant linear correlations with LIC (r = 0.787 and P < 0.001, and r = 0.859 and P < 0.001, respectively). Correlation analysis across R1ρ, R2, and R* also suggested that the correlation strength between R1ρ and R2 and between R1ρ and R* showed an increasing trend with increase in FSL.ConclusionIn this study, a strong association was observed between R1ρ and LIC at high FSLs further confirming previous findings. The results demonstrated that R1ρ at high FSL might serve as a complementary imaging biomarker for liver iron overload quantification.     

Detection of colorectal metastases in patients being treated with chemotherapy utilising SPIO-MRI: a radiological–pathological study

Objective

Chemotherapy commonly causes liver injury through sinusoidal obstructive syndrome and steatosis. Chemotherapy-induced liver injury may make it more difficult to detect metastases secondary to reduced contrast between the injured liver and metastases. The aim of this study was to determine the sensitivity of superparamagnetic iron oxide (SPIO) contrast-enhanced imaging in patients who have undergone chemotherapy prior to liver surgery.

Methods

Local ethics committee approval was obtained. Thirty-one patients with hepatic metastases completing preoperative chemotherapy were prospectively recruited. Images were reviewed independently by two blinded observers who identified and localized lesions with a four-point confidence scale. The alternative free-response receiver operator characteristic method was used to analyze the results.

Results

The sensitivity in detecting colorectal metastases following chemotherapy was 78% and 76%, respectively, for observers 1 and 2 (95% confidence interval: 71%–85% and 68%–82%). The areas under the alternative free-response receiver operator curves were 0.73 and 0.80 for observers 1 and 2, respectively.

Conclusion

Compared to previously published work on chemotherapy-naïve patients, it is clear that the sensitivity of SPIO-enhanced magnetic resonance imaging (MRI) in detecting colorectal metastases following chemotherapy is reduced. It is therefore critical that all imaging — pre-, during and postchemotherapy — is reviewed when reporting liver MRI prior to surgery.     

15Nmethacetin test to control liver function of infants with formerly very low birth weight

Abstract

Hepatic detoxification capacities of three groups of infants aged about two were estimated using the [¹⁵N]methacetin elimination test, as well as standard serum parameters:

1. Formerly hypotrophically born infants still too small for their age (n = 23)

2. Patients suffering from severe liver diseases (n = 15)

3. Patients without liver diseases (n = 14).

17 of the infants of group 1 showed ¹⁵N elimination rates as low as the rates of the liver-diseased patients of group 2. Compared to the infants of group 3, who had normal values, the findings reflect diminished hepatic monooxygenases activities in groups 1 and 2. On the other hand, the serum parameters of the infants of group 1 did not deviate from normal values estimated in group 3. Here only group 2 showed pathological values. Consequently, the [¹⁵N]methacetin test seems to be more sensitive in controlling hepatic parameters of growth retardations than the usual serum parameters used here. Further investigations have to answer the question to which proportion intrauterine malnutrition and postnatal effects, such as environmental living conditions, contribute to the retardation of liver function development in combination with growth retardation.     

The natural history of streptozotocin-stimulated non-alcoholic steatohepatitis mice followed by Gd-EOB-DTPA-enhanced MRI: Comparison with simple steatosis mice

PurposeTo clarify the development of HCC, temporal change of steatosis and Gd-EOB-DTPA enhancement of non-alcoholic steatohepatitis (NASH) model mice by magnetic resonance imaging (MRI).Materials and methodsAll animal experiments were approved by the institution's Animal Research Committee. MRI was performed on six NASH and six simple steatosis (SS) model mice every 2 weeks from the ages of 8 weeks to 16 weeks. The sequential changes in the number and size of the focal liver lesions detected on Gd-EOB-DTPA-enhanced MRI were evaluated. Additionally, the hepatic fat fraction (HFF), contrast-to-noise ratio (CNR) and relative enhancement (RE) were calculated at each time point. The temporal changes and correlations in these parameters were evaluated.ResultsAll alive NASH model mice demonstrated focal liver lesions from week 10, at the latest. Number of the lesions increased with time, and all the lesion enlarged with time. All the lesions larger than 1 mm were confirmed as hepatocellular carcinoma (HCC) pathologically. While the HFF remained constant in NASH model mice, HFF in SS model mice dramatically increased with time. CNR of the NASH model mice remained constant through the study period, while CNR in SS model mice decreased with time. Although no correlation was seen in NASH model mice, the HFF showed a negative correlation against CNR and RE in SS model mice.ConclusionDevelopment of HCC was observed using Gd-EOB-DTPA-enhanced MRI only in NASH model mice. Degree of steatosis and hepatic enhancement by Gd-EOB-DTPA was both constant in NASH model mice, while steatosis increased and hepatic enhancement decreased with time in SS model mice.     

MRI of hepatic lymphoma

Thirteen patients with biopsy proven hepatic lymphoma (2 Hodgkin, 11 Non-Hodgkin) and a control group of 15 patients with hepatic metastases were analyzed quantitatively and qualitatively by MRI. Focal hepatic lymphoma was most reliably detected (eight of eight patients) and appeared hypointense relative to liver on T₁ weighted (CNR − 7.4 ± 2.3) and hyperintense on T₂ weighted (CNR + 8.4 ± 2.9) images. The mean T₁ and T₂ relaxation times of focal hepatic lymphoma (T₁ = 832 ± 234 msec, T₂ = 84 ± 16 ms) differed significantly from adjacent non-tumorous liver (T₁ = 420 ± 121 ms, T₂ = 51 ± 9 ms; p < 0.05), however CNR values and relaxation times were similar to those of hepatic metastases. Diffuse hepatic lymphoma (microscopic periportal infiltration) was undetectable by MRI in three patients by either morphologic features or quantitative criteria. A mixed pattern of hepatic lymphoma (focal lesions and diffuse infiltration) showed focal areas of slightly decreased signal intensity on T₁ weighted images (CNR = −1.7 ± 0.4) while T₂ weighted images revealed multiple regions of focal hyperintensity (CNR = +13.3 ± 8.4) superimposed on a diffusely hyperintense liver. Our experience demonstrates that either T₁ or T₂ weighted techniques are useful in detecting focal and that T₂ weighted techniques are useful in detecting mixed hepatic lymphoma. Conventional image derived relaxation time measurements and quantitative parameters were of no additional diagnostic value.