Serum and erythrocyte membrane fatty acid levels may be used as biomarkers to assess the severity of NAFLD

The aim of this work is to study the possibility of using blood serum (BS) fatty acids (FA) and erythrocytes (ER) as diagnostic markers of the severity of NAFLD. Materials and methods. We examined 52 patients with NAFLD (51.8 ± 3.9 years), confirmed by the NLFS index, and 20 apparently healthy men (49.2 ± 4.5 years). The degree of liver fibrosis was established by indirect elastometry (FibroScan® 502 Echosens, France). 27 patients had an initial degree of fibrosis (F0-1), 25 had severe fibrosis (F2-4). The study of the composition of fatty acids of Er and BS was carried out using a GC / MS system based on three Agilent 7000B quadrupoles (USA). Results. Significant differences in the levels of fatty acids in blood serum and erythrocyte membranes in patients with NAFLD were revealed, associated with the degree of fibrosis and necroinflammatory activity. To distinguish between mild and severe fibrosis in NAFLD, the levels of saturated fatty acids (myristic, pentadecane, margarine) and omega-3 PUFAs (eicosapentaenoic, docosapentaenoic, docosahexaenoic) were found to be significant (p = 0.002-0.0003). Saturated and monounsaturated FAs (palmitelaidic, palmitoleic, vaccenic) played a key role in differentiating the degree of necroinflammatory activity (minimal versus pronounced) (p = 0.03-0.005). The created diagnostic panels (FA of blood serum and erythrocyte membranes) made it possible to differentiate patients with NAFLD with varying degrees of fibrosis. Correlations of FA levels in erythrocyte membranes and blood serum with manifestations of metabolic syndrome, indicators of liver damage in patients with NAFLD were revealed. Conclusions. The established differences in fatty acid profiles of blood serum and erythrocyte membranes in patients with NAFLD, associated with the degree of fibrosis, necroinflammatory activity, manifestations of metabolic syndrome and indicators of liver damage, should be considered as promising biomarkers for assessing the severity of NAFLD.

The usefulness of noninvasive liver stiffness assessment using shear-wave elastography for predicting liver fibrosis in children

Background Pediatric patients with liver disease require noninvasive monitoring to evaluate the risk of fibrosis progression. This study aimed to identify the significant factors affecting liver stiffness values using two-dimensional shear-wave elastography (2D-SWE), and determine whether liver stiffness can predict the fibrosis stage of various childhood liver diseases. Methods This study included 30 children (22 boys and 8 girls; mean age, 5.1 ± 6.1 years; range, 7 days–17.9 years) who had undergone biochemical evaluation, 2D-SWE examination, histopathologic analysis of fibrosis grade (F0 to F3), assessment of necroinflammatory activity, and steatosis grading between August 2016 and March 2020. The liver stiffness from 2D-SWE was compared between fibrosis stages using Kruskal–Wallis analysis. Factors that significantly affected liver stiffness were evaluated using univariate and multivariate linear regression analyses. The diagnostic performance was determined from the area under the receiver operating curve (AUC) values of 2D-SWE liver stiffness. Results Liver stiffness at the F0-1, F2, and F3 stages were 7.9, 13.2, and 21.7 kPa, respectively (P < 0.001). Both fibrosis stage and necroinflammatory grade were significantly associated with liver stiffness (P < 0.001 and P = 0.021, respectively). However, in patients with alanine aminotransferase (ALT) levels below 200 IU/L, the only factor affecting liver stiffness was fibrosis stage (P = 0.030). The liver stiffness value could distinguish significant fibrosis (≥ F2) with an AUC of 0.950 (cutoff value, 11.3 kPa) and severe fibrosis (F3 stage) with an AUC of 0.924 (cutoff value, 18.1 kPa). The 2D-SWE values for differentiating significant fibrosis were 10.5 kPa (≥ F2) and 18.1 kPa (F3) in patients with ALT levels below 200 IU/L. Conclusion The liver stiffness values on 2D-SWE can be affected by both fibrosis and necroinflammatory grade and can provide excellent diagnostic performance in evaluating the fibrosis stage in various pediatric liver diseases. However, clinicians should be mindful of potential confounders, such as necroinflammatory activity or transaminase level, when performing 2D-SWE measurements for liver fibrosis staging.

Association of serum 25-hydroxyvitamin D levels with severe necroinflammatory activity and inflammatory cytokine production in type I autoimmune hepatitis

25-Hydroxyvitamin D [25(OH)D] has been reported to be associated with several chronic liver diseases. The relationship between 25(OH)D and autoimmune hepatitis (AIH) pathogenesis is incompletely understood. We investigated the association of serum total and free 25(OH)D levels with necroinflammatory activity and cytokine levels in 66 patients with AIH diagnosed in our hospital. The median age at AIH diagnosis was 57 years, and the male:female ratio was 7:59. The median serum total 25(OH)D level in therapy-naïve patients with AIH was 14.2 ng/mL (interquartile range [IQR], 11.4–17.9 ng/mL). Of the 66 patients with AIH, 36 had serum total 25(OH)D levels of < 15 ng/mL and were considered to have vitamin D deficiency, and 30 had serum total 25(OH)D levels of ≥ 15 ng/mL. Patients with acute-onset AIH had significantly lower serum total 25(OH)D levels than those with chronic-onset AIH. In particular, serum total 25(OH)D levels were significantly lower in patients with severe forms of AIH. Furthermore, the serum total 25(OH)D level was positively correlated with the serum albumin level and prothrombin time and negatively correlated with the serum total bilirubin level and necroinflammatory activity in AIH. Multivariate logistic regression analysis showed that the serum total 25(OH)D level was an independent factor for severe necroinflammatory activity. Interestingly, AIH patients with serum total 25(OH)D levels of < 15 ng/mL had higher levels of inflammatory cytokines such as interferon-γ and interleukin-33. Free 25(OH)D levels were correlated with total 25(OH)D levels, and the percentage of free 25(OH)D was significantly associated with necroinflammatory activity. In conclusion, 25(OH)D deficiency may play an important role in predicting AIH severity via inflammatory cytokine production.

Diffusion Tensor Imaging Quantifying the Severity of Chronic Hepatitis in Rats

Background: Diffusion tensor imaging (DTI) is mainly used for detecting white matter fiber in the brain. DTI was applied to assess fiber in liver disorders in previous studies. However, the data obtained have been insufficient in determining if DTI can be used to exactly stage chronic hepatitis. This study assessed the value of DTI for staging of liver fibrosis (F), necroinflammatory activity (A) and steatosis (S) with chronic hepatitis in rats. Methods: Seventy male Sprague-Dawley rats were divided into a control group(n=10) and an experimental group(n=60). The rat models of chronic hepatitis were established by abdominal subcutaneous injections of 40% CCl4. All of the rats underwent 3.0T MRI. Regions of interest (ROIs) were subjected to DTI to estimate the MR parameters (rADC value and FA value). Histopathology was used as the reference standard. Multiple linear regression was used to analyze the associations between the MR parameters and pathology. The differences in the MR parameters among the pathological stages were evaluated by MANOVA or ANOVA. The LSD test was used to test for differences between each pair of groups. ROC analysis was also performed. Results: The count of each pathology was as follows: F0(n=15), F1(n=11), F2(n=6), F3(n=9), F4(n=6); A0(n=8), A1(n=16), A2(n=16), A3(n=7); S0(n=10), S1(n=7), S2(n=3), S3(n=11), S4(n=16). The rADC value had a negative correlation with liver fibrosis (r=-0.392, P=0.008) and inflammation (r=-0.359, P=0.015). The FA value had a positive correlation with fibrosis (r=0.409, P=0.005). Significant differences were found in the FA values between F4 and F0~F3 (P=0.03), while no significant differences among F0~F3 were found (P> 0.05). The AUC of the FA value differentiating F4 from F0~F3 was 0.909 (p<0.001) with an 83.3% sensitivity and an 85.4% specificity when the FA value was at the cut-off of 588.089 (×10-6mm2/s).Conclusion: The FA value for DTI can distinguish early cirrhosis from normal, mild and moderate liver fibrosis, but the rADC value lacked the ability to differentiate among the fibrotic grades. Both the FA and rADC values were unable to discriminate the stages of necroinflammatory activity and steatosis.

SUN-506 Hyperthyroidism Induced Hepatic Apoptosis

Background: Graves’ disease is commonly associated with abnormal liver function tests, most frequently ALP, but the exact mechanism is not fully understood. In vitro and in vivo animal studies have shown elevated T3 activity can induce hepatocyte apoptosis via a mitochondrial-mediated pathway. This case demonstrates a patient with elevated aminotransferases and hepatic apoptosis most likely secondary to severe hyperthyroidism. Clinical Case: 50 year old female with a past medical history of migraines was seen by primary care for fatigue and 15 lb weight loss in one month. She was found to be hyperthyroid with TSH &lt; 0.1 uIU/L (n=0.34-5.6), free T4 3.48 ng/dL (n=0.58-1.64) and mildly elevated aminotransferases of AST 77 IU/L (n=15-41), ALT 144 IU/L (n=12-63), which increased within a week to 159 IU/L and 309 IU/L respectively. ALP and bilirubin were within normal range. She was started on methimazole 20 mg twice daily by her PCP. The patient developed vomiting and stopped taking methimazole after 3-4 days. Upon initial presentation to endocrine clinic, found to be clinically hyperthyroid and as LFTs were improved but still elevated, she was re-challenged with methimazole at a lower dose as well as started on a beta blocker and cholestyramine. TT3 checked was elevated at 2.10 ng/mL (n 0.87-1.78). Graves’ disease was confirmed with elevated TSI as well as RAI uptake and scan showing increased homogenous uptake. She had extensive workup for another etiology by hepatology including autoimmune, which were negative. Her fibrosis score was stage F1-F2 (n=F0) and necroinflammatory activity grade A3 indicating severe activity (n=grade A0). Core needle biopsy of the liver showed focal lytic necrosis/apoptosis and abundant pigment-laden Kupffer cells signifying recent hepatocellular injury. Her AST and ALT down trended and normalized with repeat fibrosis score of F1 and necroinflammatory activity grade A0. She eventually had definitive therapy with RAI treatment. Conclusion: In most cases of hyperthyroid induced liver dysfunction, liver histology showed fatty infiltration, cytoplasmic vacuolization, nuclear irregularity and hyperchromatism. This case, without any other known causes that could explain her hepatic injury, indicates the possible role of hyperthyroidism in hepatic apoptosis.