Interferon-α2a increases serum concentration of hyaluronic acid and type III procollagen aminoterminal propeptide in patients with chronic hepatitis B virus infection

Interferon-alpha (IFN-) has become an important drug for the treatment of chronic viral liver diseases. However, the action of IFN- remains unclear. We investigated whether human recombinant IFN- modulates serum concentrations of hyaluronic acid (HA) and type III procollagen aminoterminal propeptide (P-III-NP) in 56 patients with chronic hepatitis-B under IFN- therapy. IFN- increased the HA serum level in 44 of 46 patients and, after cessation of treatment, HA serum levels returned to the pretherapy levels. The increase of HA serum level was higher in patients with active cirrhosis (aC) than in patients with chronic persistent hepatitis (CPH) and in patients with severe inflammation compared to those with moderate inflammation. Interestingly, HA serum concentration was unrelated to IFN dose and was of no predictive value for therapy response. In contrast, IFN- increased significantly the P-III-NP serum level in patients with aC only. During follow-up, P-III-NP serum level decreased late in responders in parallel to the decrease of serum level of liver enzymes, in non-responders it was without significant change. The first dose of IFN induced a significant increase in HA serum level in each of 10 patients but in none of four healthy volunteers. In contrast, P-III-NP serum concentrations were not influenced by the first IFN- dose. We conclude that: (1) immunstimulation with IFN- induces a rapid increase of HA serum level in patients with chronic hepatitis B but not in normal persons; (2) IFN- increases P-III-NP serum level only in patients with active liver cirrhosis; (3) measurement of HA and P-III-NP serum levels does not help predict response to IFN-, and (4) HA serum level may be used as a compliance indicator.     

Serum bile acids in cystic fibrosis patients – glycodeoxycholic acid as a potential marker of liver disease

BackgroundCystic fibrosis (CF) and CF-related liver disease can lead to disturbances in bile acid metabolism.AimThis study determined serum bile acid concentrations in CF to define their usefulness in liver disease assessment.MethodsPrimary, secondary and conjugated bile acid levels were measured in three CF groups (25 patients each) exhibiting: liver cirrhosis, other liver disease, no liver disease, and in 25 healthy subjects (HS).ResultsBile acid levels were higher in CF patients than in HS, except for glycodeoxycholic acid (GDCA). However, bile acid concentrations did not differ between patients with cirrhosis and other liver involvement. GDCA and deoxycholic acid (DCA) differentiated CF patients with non-cirrhotic liver disease from those without liver disease (GDCA-AUC: 0.924, 95%CI 0.822–1.000, p<0.001; DCA-AUC: 0.867, 95%CI: 0.731–1.000, p<0.001). Principal component analysis revealed that in CF liver disease was related to GDCA, GGTP activity, severe genotype and pancreatic insufficiency.ConclusionsA CF-specific bile acid profile was defined and shown to relate to liver disease. GDCA differentiates patients with non-cirrhotic liver involvement from those with no detectable liver disease. Hence, GDCA is a candidate for validation as a biomarker of non-cirrhotic progression of liver disease in CF.     

Dose–response relationship of serum uric acid levels with risk of stroke mortality

Introduction

Results from the recent meta-analysis suggested that higher serum uric acid (SUA) levels are positively associated with risk of stroke. However, the relationship of SUA levels with risk of stroke is still unclear.

Materials and methods

Data from prospective cohort studies on SUA levels and risk of stroke mortality was used. Dose–response relationship was assessed by restricted cubic spline model and multivariate random effect meta-regression.

Results

A non-linear relationship (P_{for non-linearity} = 0.004) of SUA levels with risk of stroke mortality was found for men, and the relative risk (RR) with 95% confidence interval (CI) of stroke mortality was 1.00 (0.99–1.01), 0.99 (0.94–1.04), 0.98 (0.91–1.06), 1.00 (0.90–1.12), 1.17 (1.09–1.24) and 1.52 (1.33–1.78) for 2, 3, 4, 5, 6 and 7 mg/dL of SUA levels, respectively. For women, the departure from linearity was not significant (P_{for non-linearity} = 0.67), and the RR (95 %CI) of stroke mortality was 1.02 (0.99–1.04), 1.10 (0.97–1.20), 1.15 (0.96–1.37), 1.25 (1.09–1.44), 1.39 (1.28–1.50) for 2, 3, 4, 5, 6 mg/dL of SUA levels, respectively.

Conclusions

Different dose–response relationships of SUA levels with risk of stroke mortality might exist for men and for women. Dose–response relationship of SUA levels with risk of stroke incidence needs to be explored.     

Can the MEGX test and serum bile acids improve the prognostic ability of Child-Pugh's score in liver cirrhosis?

BACKGROUND: Liver transplantation is nowadays the therapeutic option for end-stage liver disease. Correct disease staging is the main step towards improving the timing of listing for liver transplantation so as to avoid premature or late entry. The need for correct prognostic evaluation is due to the limited number of donors and to the increasing number of patients awaiting transplantation. Our aim was to verify whether Child-Pugh's score might be improved by adding the monoethylglycinexylidide (MEGX) formation test and/or serum bile acid determination. METHODS: We evaluated 182 cirrhotic patients (44 Child-Pugh class A, 97 class B, and 41 class C) of mixed aetiology referring to a tertiary care centre for functional staging of liver disease. These patients were prospectively followed-up for 12-72 months. During this period, 45 patients died, 46 received a transplant, and 91 survived without transplantation. The end-point of analysis was either survival or liver disease-related death at the 6th, 12th, 18th and 24th months of follow-up. The 46 transplanted patients were excluded from the study upon transplantation. RESULTS: In our study, a cut-off for Child-Pugh's score < 8 confirmed its usefulness, especially in short-term prognostic prediction, while mid- and long-term prediction improved by almost 10% by using the combination of a Child- Pugh's score > 8 and an MEGX value < 15 mg/l. Cox's multi-variate regression analysis indicated that MEGX values either with Child-Pugh's score or with prothrombin activity and ascites were independent prognostic variables. CONCLUSIONS: Besides confirming that Child-Pugh's score as the basis of prognostic evaluation of cirrhotic patients, these results suggest that the MEGX test might be a complement to the original score when a patient is being evaluated for a liver transplantation programme.     

Effects of bile salts on plasma concentration of β-endorphin-like immunoreactivity in men

The effects of bile salts on the release of -endorphin-like immunoreactivity ( -END-LI) were investigated in men using a specific radioimmunoassay developed by the authors. Plasma -END-LI was determined after extraction by the acid-acetone method (recovery: 73±5%). Oral administration of 400 mg of sodium taurocholate caused a rise in plasma -END-LI from 9.9±0.5 pmol/liter to 21.3±1.2 pmol/liter after 30 min and 18.1 ±0.5 pmol/liter after 60 min, with return to the initial value after 90 min. Oral administration of chenodeoxycholic acid (CDCA) also increased plasma -END-LI from a basal level of 8.4±0.7 pmol/liter to 18.7±0.8 pmol/liter after 30 min. Oral administration of ursodeoxycholic acid (UDCA) increased plasma -END-LI from 7.3±0.3 pmol/liter to 30.6±0.2 pmol/liter after 30 min. In gel chromatography, the -END-LI released after UDCA administration separated into two components, which eluted in the same positions as human -lipotropin and human -endorphin, respectively. These results suggested that bile salts may participate the release of -END-LI.     

Overexpression of cholesterol 7α-hydroxylase promotes hepatic bile acid synthesis and secretion and maintains cholesterol homeostasis

We reported previously that mice overexpressing cytochrome P450 7a1 (Cyp7a1; Cyp7a1-tg mice) are protected against high fat diet-induced hypercholesterolemia, obesity, and insulin resistance. Here, we investigated the underlying mechanism of bile acid signaling in maintaining cholesterol homeostasis in Cyp7a1-tg mice. Cyp7a1-tg mice had two-fold higher Cyp7a1 activity and bile acid pool than did wild-type mice. Gallbladder bile acid composition changed from predominantly cholic acid (57%) in wild-type to chenodeoxycholic acid (54%) in Cyp7a1-tg mice. Cyp7a1-tg mice had higher biliary and fecal cholesterol and bile acid secretion rates than did wild-type mice. Surprisingly, hepatic de novo cholesterol synthesis was markedly induced in Cyp7a1-tg mice but intestine fractional cholesterol absorption in Cyp7a1-tg mice remained the same as wild-type mice despite the presence of increased intestine bile acids. Interestingly, hepatic but not intestinal expression of several cholesterol (adenosine triphosphate-binding cassette G5/G8 [ABCG5/G8], scavenger receptor class B, member 1) and bile acid (ABCB11) transporters were significantly induced in Cyp7a1-tg mice. Treatment of mouse or human hepatocytes with a farnesoid X receptor (FXR) agonist GW4064 or bile acids induced hepatic Abcg5/g8 expression. A functional FXR binding site was identified in the Abcg5 gene promoter. Study of tissue-specific Fxr knockout mice demonstrated that loss of the Fxr gene in the liver attenuated bile acid induction of hepatic Abcg5/g8 and gallbladder cholesterol content, suggesting a role of FXR in the regulation of cholesterol transport. CONCLUSION: This study revealed a new mechanism by which increased Cyp7a1 activity expands the hydrophobic bile acid pool, stimulating hepatic cholesterol synthesis and biliary cholesterol secretion without increasing intestinal cholesterol absorption. This study demonstrated that Cyp7a1 plays a critical role in maintaining cholesterol homeostasis and underscores the importance of bile acid signaling in regulating overall cholesterol homeostasis.     

Measurement of hepatic functional mass by means of ^13C-methacetin and 13C-phenylalanine breath tests in chronic liver disease： Comparison with Child-Pugh score and serum bile acid levels

AIM: To evaluate and compare the clinical usefulness of ~(13)C-phenylalanine and ~(13)C-methacetin breath tests in quantitating functional hepatic mass in patients with chronic liver disease and to further compare these results with those of conventional tests, Child-Pugh score and serum bile acid levels. METHODS: One hundred and forty patients (50 HCV-related chronic hepatitis, 90 liver cirrhosis patients) and 40 matched healthy controls were studied. Both breath test and routine liver test, serum levels of cholic and chenodeoxycholic acid conjugates were evaluated. RESULTS: Methacetin breath test, expressed as 60 min cumulative percent of oxidation, discriminated the hepatic functional capacity not only between controls and liver disease patients, but also between different categories of chronic liver disease patients. Methacetin breath test was correlated with liver function tests and serum bile acids. Furthermore, methacetin breath test, as well as serum bile acids, were highly predictive of Child-Pugh scores. The diagnostic power of phenylalanine breath test was always less than that of methacetin breath test. CONCLUSION: Methacetin breath test represents a safe and accurate diagnostic tool in the evaluation of hepatic functional mass in chronic liver disease patients.     

Clinical analysis of patients with iatrogenic bile duct injury

BACKGROUND: The main cause of iatrogenic bile duct injury is misidentification of the common bile duct as the cystic duct. In this article, we summarize the experience in the treatment of 112 patients with iatrogenic bile duct injury. METHOD: Clinical data of these patients treated at 10 hospitals of Songhua river area, Heilongjiang province, China from January 1978 to January 2005 were analyzed retrospectively. RESULTS: In 55.4% patients (62/112), iatrogenic bile duct injury was due to misidentification of the anatomy of Calot's triangle before cholecystectomy. Their diagnosis was based on clinical features, celiac puncture and imaging examination in which ultrasonography was most sensitive, giving a diagnostic rate of 97. 5%. Six types of injury were identified according to their locations, and type Ⅲ damage was commonly seen ( 92/112 ). The curative rate in this group was 95.5% (107/112). Eighty-seven patients (77.7%) underwent Roux-en-Y choledochojejunostomy with a cure rate of 94.3% (82/87). CONCLUSION: The prevention of iatrogenic bile duct injury lies in identifying the topography of extrahepatic bile ducts. Roux-en-Y choledochojejunostomy is usually the treatment of choice.     

Ventricular K currents are reduced in mice with elevated levels of serum TNFα

In the present study mice were treated with tumor necrosis factor alpha (TNFα) for 6 weeks to determine if chronic TNFα treatment could produce serum levels of TNFα similar to what has been observed in disease states (heart failure, HIV) and to determine if these levels of TNFα alter ventricular K⁺ currents. Mice chronically treated with TNFα and sham treated mice were utilized for experiments. Serum levels were measured with a Searchlight® protein array. Patch-clamp techniques, real-time PCR and Western blot analysis were used to study K⁺ current densities and K⁺ channel expression. Results showed that serum concentrations of TNFα were significantly higher in TNFα treated mice compared to controls (control: 9.5 ± 1.5 pg/ml, TNFα: 27.4 ± 5.0 pg/ml; p < 0.05) and comparable to serum TNFα levels observed in heart failure and HIV models. In ventricular myocytes from TNFα treated mice the outward K⁺ currents I_to and I_Kur were significantly reduced (at + 30 mV: I_to: control: 45.0 ± 2.9 pA/pF, TNFα: 34.5 ± 2.9 pA/pF; p < 0.05; I_Kur: control 34.1 ± 2.7 pA/pF, TNFα: 25.0 ± 2.2 pA/pF; p < 0.05). Expression studies revealed that ventricular mRNA and protein expression for the channels underlying I_to and I_Kur did not differ between the two groups. However, the recovery from inactivation for I_Kur was significantly longer in TNFα treated mice. Overall, this study shows that pathologically relevant levels of serum TNFα modulate K⁺ currents in mouse ventricle. These findings could help to explain the role of TNFα in the pathogenesis of cardiac arrhythmia.     

Efficacy of ursodeoxycholic acid therapy in chronic viral hepatitis C with high serum γ-glutamyltranspeptidase levels

We administered ursodeoxycholic acid (UDCA) orally, at a daily dose of 600 mg, for 4 months to 36 patients with chronic viral hepatitis C. Another 36 patients with chronic viral hepatitis C, treated with placebo for 4 months, served as controls. None of the patients were alcoholics and none suffering from auto-immune hepatitis. Of the 36 patients in the UDCA-treated group, 13 had high levels of serum -glutamyl-transpeptidase (GGT), i.e., exceeding 150U/l (normal <50U/l). Histological examination of liver biopsy specimens obtained from 10 patients in this group before treatment suggested that damage of the interlobular bile ducts was prominent in patients with higher levels of serum GGT. After 1 month of UDCA treatment, significant decreases in the levels of serum GGT, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were observed (P<0.05 for GGT and AST), and the decreases continued for the 4-month treatment period. The reduction of GGT levels was the most prominent change in the liver function indices; the percent change in the GGT level was –25.2±4.4 (mean percent change ± SE) at 1 month and –38.0±5.0 at 4 months. A significant correlation was observed between the serum AGGT level (GGT value before treatment minus value after 3 months of treatment) and the total score for morphological injury of the bile ducts (P<0.05). These results suggested that UDCA has the potential to reverse hepatocellular damage in patients with chronic viral hepatitis C, in whom high GGT levels may be due, in part, to a damaged interlobular bile duct. UDCA may be useful for the treatment of chronic viral hepatitis C, especially in patients exhibiting a high level of GGT.     

Analysis of organic acids after incubation with (16-2H3)palmitic acid in fibroblasts from patients with mitochondrial β-oxidation defects

The analysis of acylcarnitines as products of incubation of intact fibroblasts with isotope-labelled precursors, usually (16-(2)H(3))hexadecanoic acid, is an advanced in vitro method for the study of mitochondrial beta-oxidation defects. We propose a technique for the measurement of the organic acid intermediates after hydrolysis of the acylcarnitines using electron-impact gas chromatography-mass spectrometry. For some mitochondrial beta-oxidation deficiencies, the characteristic profile enables us to approach the diagnosis with clear differentiation.     

Experimental study on the relationship of bile acids and myocardium damage in obstructive jaundice

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Bile acid-CoA ligase deficiency—a new inborn error of bile acid metabolism

Born at 27 weeks gestation, a child of consanguineous parents of Pakistani origin required prolonged parenteral nutrition. She developed jaundice, with extensive fibrosis and architectural distortion at liver biopsy; jaundice resolved with supportive care. Serum γ-glutamyl transpeptidase values were within normal ranges. The bile acids in her plasma and urine were >85% unconjugated (non-amidated). Two genes encoding bile-acid amidation enzymes were sequenced. No mutations were found in BAAT, encoding bile acid-CoA : aminoacid N-acyl transferase. The patient was homozygous for the missense mutation c.1012C > T in SLC27A5, predicted to alter a highly conserved amino-acid residue (p.H338Y) in bile acid-CoA ligase (BACL). She also was homozygous for the missense mutation c.1772A > G in ABCB11, predicted to alter a highly conserved amino-acid residue (p.N591S) in bile salt export pump (BSEP). BACL is essential for reconjugation of bile acids deconjugated by gut bacteria, and BSEP is essential for hepatocyte-canaliculus export of conjugated bile acids. A female sibling born at term had the same bile-acid phenotype and SLC27A5 genotype, without clinical liver disease. She was heterozygous for the c.1772A > G ABCB11 mutation. This is the first report of a mutation in SLC27A5. The amidation defect may have contributed to cholestatic liver disease in the setting of prematurity, parenteral nutrition, and homozygosity for an ABCB11 mutation.