IP Library › Granted Patent US 12,332,255
Granted Patent B2
US 12,332,255 · App. 17/504,788 · Granted Jun 17, 2025

Methods for monitoring treatment of chronic liver disease

Inventors: Gregory Thomas Everson (Englewood, CO); Steve Mark Helmke (Denver, CO)
Assignee: The Regents of the University of Colorado
G01N33/92A61K31/575G16H50/20G16H50/30G01N2800/085
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Quick Facts
Patent No.
US 12,332,255
App. No.
17/504,788
Granted
Jun 17, 2025
Kind
B2
Abstract

A Disease Severity Index (DSI) is provided for assessment of chronic liver disease in a patient using non-invasive liver function test results. A DSI was derived from non-invasive liver function test results based on hepatic blood flow. The DSI is used in methods for prediction of clinical outcomes, prediction of response to antiviral treatment, and assessment of progression of chronic liver diseases. Non-invasive methods to diagnose three distinct categories of patients with Primary Sclerosing Cholangitis (PSC) are provided. The methods can be used to diagnose PSC patients as Slow Progressors, Moderate Progressors and Rapid Progressors.

Claims (67)

1. A method of predicting a response to a treatment of a chronic liver disease in a patient, comprising

determining a baseline Disease Severity Index (DSI) value in the patient having a chronic liver disease prior to the treatment; and

comparing the baseline DSI value in the patient to a DSI cutoff value, wherein a baseline DSI value in the patient below the DSI cutoff value indicates a higher predicted response rate to the treatment in the patient compared to a subject exhibiting a DSI value above the DSI cutoff value,

wherein the determining of the baseline DSI value in the patient comprises

performing a cholate liver function tests in the patient to obtain a portal hepatic filtration rate (portal HFR) value, a systemic hepatic filtration rate (systemic HFR) value, and optionally a cholate SHUNT test value; and

deriving the disease severity index (DSI) values from the cholate test results,

wherein the portal HFR test value in the patient is determined by a method comprising:

(a) receiving a plurality of blood or serum samples collected from the patient having a chronic liver disease, following oral administration of a dose of a first distinguishable cholate compound (dose oral ) to the patient, wherein the samples have been collected from the patient over intervals of from two to seven time points spanning a period of time of no more than about 90 minutes after administration;

(b) measuring concentration of the first distinguishable cholate compound in each sample comprising HPLC-MS;

(c) generating an individualized oral clearance curve from the concentration of the first distinguishable cholate in each sample comprising using a computer algorithm curve fitting to a model distinguishable cholate clearance curve;

(d) computing the area under the individualized oral clearance curve (AUC) (mg/mL/min) and dividing the dose (in mg) by AUC of the orally administered first distinguishable cholate compound to obtain the oral cholate compound clearance in the patient; and

(e) dividing the oral cholate compound clearance by the weight of the patient in kg to obtain the portal HFR value in the patient (mL/min/kg); or,

wherein the portal HFR value in the patient is estimated from a cholate STAT test value in the patient.

2. The method of claim 1 , wherein the cutoff value was derived from DSI values in one or more normal healthy subjects, group of known subjects, or within the patient over time.

3. The method of claim 2 , wherein the group of known subjects is selected from a group of healthy subjects, a group of subjects with chronic liver disease having a fibrosis stage;

portal hypertension; Childs-Turcotte-Pugh (CTP) score A; CTP score B; CTP score C; Model for End-stage Liver Disease (MELD) score; primary sclerosing cholangitis (PSC) not listed for transplant; PSC listed for liver transplant; PSC listed for liver transplant without varices; PSC listed for liver transplant with varices; ascites; stomal bleeding; splenomegaly; varices; variceal hemorrhage; hepatic encephalopathy; decompensation; or liver related death.

4. The method of claim 3 , wherein the fibrosis stage is determined by a method selected from the group consisting of liver biopsy; elastography; fibrosis serum biomarker; circulating proteins or fragments thereof; and nonalcoholic fatty liver disease (NAFLD) fibrosis score.

5. The method of claim 4 , wherein the liver biopsy determines Ishak fibrosis score (liver biopsy) of F2 (mild portal fibrosis), F3, F4 (moderate bridging fibrosis), F5 (nodular formation and incomplete cirrhosis), or F6 (cirrhosis).

6. The method of claim 3 , wherein the DSI cutoff value is selected from the group consisting of 14, 15, 18, 19, 20, 23, 25, 30, 35, 36, and 40.

7. The method of claim 1 , wherein the chronic liver disease is selected from the group consisting of chronic hepatitis C (CHC), chronic hepatitis B, alcoholic liver disease, Alcoholic SteatoHepatitis (ASH), Non-Alcoholic Fatty Liver Disease (NAFLD), steatosis, Non-Alcoholic SteatoHepatitis (NASH), autoimmune liver disease, cryptogenic cirrhosis, hemochromatosis, Wilson's disease, alpha-1-antitrypsin deficiency, liver cancer, liver failure, cirrhosis, primary sclerosing cholangitis (PSC), and other cholestatic liver diseases.

8. The method of claim 7 , wherein the chronic liver disease is selected from the group consisting of CHC, PSC, and NASH.

9. The method of claim 1 , wherein the treatment is selected from the group consisting of antiviral treatment, antifibrotic treatment, antibiotics, immunosuppressive treatments, anti-cancer treatments, ursodeoxycholic acid, insulin sensitizing agents, interventional treatment, liver transplant, lifestyle changes, dietary restrictions, low glycemic index diet, antioxidants, vitamin supplements, transjugular intrahepatic portosystemic shunt (TIPS), catheter-directed thrombolysis, balloon dilation and stent placement, balloon-dilation and drainage, weight loss, exercise, and avoidance of alcohol.

10. The method of claim 1 , wherein the portal HFR value, systemic HFR value, and/or SHUNT value are obtained on the same test day.

11. The method of claim 1 , wherein the systemic HFR value in the patient is determined by a method comprising

(a) receiving a plurality of blood or serum samples collected from a patient having or at risk of a chronic liver disease, following intravenous administration of a dose of a second distinguishable cholate compound (dose iv ) to the patient, wherein the samples have been collected from the patient over intervals of from two to seven time points spanning a period of time of no more than about 90 minutes after administration;

(b) measuring concentration of the second distinguishable cholate compound in each sample comprising HPLC-MS;

(c) generating an individualized intravenous clearance curve from the concentration of the second distinguishable cholate compound in each sample comprising using a computer algorithm curve fitting to a model second distinguishable cholate clearance curve;

(d) computing the area under the individualized intravenous clearance curve (AUC) (mg/mL/min) and dividing the dose (in mg) by AUC of the intravenously administered second distinguishable cholate compound to obtain the intravenous cholate compound clearance in the patient; and

(e) dividing the intravenous cholate clearance by the weight of the patient in kg to obtain the systemic HFR value in the patient (mL/min/kg).

12. The method of claim 1 , wherein the deriving of the baseline DSI value comprises

employing a disease severity index equation (DSI equation) to obtain a DSI value or mathematically transformed DSI value in the patient, wherein the DSI equation comprises one or more portal HFR or mathematically transformed portal HFR terms, one or more systemic HFR or mathematically derived systemic HFR terms, and a constant to obtain the baseline DSI value.

13. The method of claim 12 , wherein the disease severity index (DSI) equation comprises

DSI=A (SHUNT)+ B (log e portal HFR )+ C (log e systemic HFR )+ D

wherein

SHUNT is SHUNT test value in the patient (%);

portal HFR is portal hepatic flow rate (HFR) test value in the patient as mL/min/kg, wherein kg is body weight of the patient;

systemic HFR is systemic HFR value in the patient as mL/min/kg, wherein kg is body weight of the patient;

A is a SHUNT coefficient;

B is a Portal HFR coefficient;

C is a Systemic HFR coefficient; and

D is the constant.

14. The method of claim 13 , wherein the SHUNT coefficient A is a number from 0 to positive 25; the Portal HFR coefficient B is a number from 0 to negative 25; and the Systemic HFR coefficient C is a number from 0 to negative 25, and wherein at least one of A, B, and C is not 0.

15. The method of claim 13 , wherein the constant D is a positive number between 5 and 125.

16. The method of claim 13 , wherein the SHUNT test value in the patient is determined by a method comprising

(a) receiving a plurality of blood or serum samples collected at two to seven time points over no more than about 90 min from the patient having a chronic liver disease, following oral administration of a dose of a first distinguishable cholate compound (dose oral ) to the patient and simultaneous intravenous co-administration of a dose of a second distinguishable cholate compound (dose iv ) to the patient, wherein the samples have been collected over intervals spanning a period of time after administration;

(b) quantifying the concentration of the first and the second distinguishable cholate compounds in each sample comprising HPLC-MS;

(c) generating an individualized oral clearance curve from the concentration of the first distinguishable cholate compound in each sample comprising using a computer algorithm curve fitting to a model oral distinguishable cholate clearance curve and computing the area under the individualized oral clearance curve (AUCoral);

(d) generating an individualized intravenous clearance curve from the concentration of the second distinguishable cholate compound in each sample by use of a computer algorithm curve fitting to a model intravenous second distinguishable cholate clearance curve and computing the area under the individualized intravenous clearance curve (AUCiv); and

(e) calculating the shunt value in the patient using the formula:

AUC oral /AUC iv ×Dose iv /Dose oral ×100%.

17. The method of claim 16 , wherein the first distinguishable cholate compound is a first stable isotope labeled cholate and the second distinguishable cholate compound is a second stable isotope labeled cholate.

18. The method of claim 17 , wherein the first and second stable isotope labeled cholates are selected from 2,2,4,4-d4 cholate and 24- 13 C-cholate.

19. The method of claim 16 , wherein the samples have been collected from the patient at time points comprising about 0, about 5, about 20, about 45, about 60, and about 90 minutes after the administration.

20. The method of claim 1 , further comprising performing an additional test or procedure selected from the group consisting of liver ultrasound monitoring; elastography; liver biopsy; histological assessment; biomarker or biomarker panel; metabolic test; Model for End Stage Liver Disease (MELD) score; and Childs-Turcotte-Pugh (CTP) score.

21. The method of claim 20 , wherein the biomarker or biomarker panel is a clinical biochemistry laboratory assay selected from the group consisting of serum albumin; alanine transaminase; aspartate transaminase; alkaline phosphatase; total bilirubin; direct bilirubin; creatinine; gamma glutamyl transpeptidase; complete blood count (CBC); platelet count; hematocrit; 5′ Nucleotidase; and prothrombin time-international normalized ratio (PT-INR).

22. The method of claim 20 , wherein the biomarker or biomarker panel is a fibrosis test selected from the group consisting of fibrosis serum biomarker; circulating proteins or fragments thereof; Ishak fibrosis score; fibrotest biochemical markers score; and nonalcoholic fatty liver disease (NAFLD) fibrosis score.

23. The method of claim 20 , wherein the biomarker or biomarker panel comprises a pattern of pro-inflammatory cytokines.

24. The method of claim 23 , wherein the pro-inflammatory cytokines are selected from the group consisting of IFN-γ, TNF-α, GM-CSF, IL-1β, IL-6, and IL-8.

25. The method of claim 1 , wherein the STAT test value in the patient is obtained by a method comprising

(a) receiving a single blood or serum sample collected from the patient, following oral administration of a dose of a distinguishable cholate compound (dose oral ), wherein the sample has been collected from the patient at a specific time point within 20-180 minutes after administration; and

(b) measuring concentration of the distinguishable cholate compound (uM) in the sample.

26. The method of claim 25 , further comprising

(c) transforming the distinguishable cholate compound concentration (uM) into an estimated portal flow (mL/min/kg) comprising

y= 0.9702 x+ 0.0206, wherein

x is the LOG Portal HFR (FLOW);

y is LOG STAT; and

kg is body weight of patient.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 19, 2021
From: EVERSON, GREGORY THOMAS; HELMKE, STEVE MARK
To: THE REGENTS OF THE UNIVERSITY OF COLORADO, A BODY CORPORATE
Reel/Frame 057833/0692 →
Continuity (6)
Division 16460767 · Jul 2, 2019
Continuation 15673191 · Aug 9, 2017
Continuation 14729987 · Jun 3, 2015
Continuation 14078058 · Nov 12, 2013
Provisional Application 61725292 · Nov 12, 2012
Related Publication 20220034915A1 · Feb 3, 2022
References Cited (97)
US 4207308A · Spenney · 1980 [cited by applicant]
US 6778269B2 · Fink et al. · 2004 [cited by applicant]
US 7060250B2 · McMurry et al. · 2006 [cited by applicant]
US 8613904B2 · Everson et al. · 2013 [cited by applicant]
US 8778299B2 · Everson · 2014 [cited by applicant]
US 8961925B2 · Everson et al. · 2015 [cited by applicant]
US 9091701B2 · Everson et al. · 2015 [cited by applicant]
US 9417230B2 · Everson · 2016 [cited by applicant]
US 9759731B2 · Everson et al. · 2017 [cited by applicant]
US 10520517B2 · Everson et al. · 2019 [cited by applicant]
US 11181536B2 · Everson et al. · 2021 [cited by applicant]
US 20060067881A1 · Groman et al. · 2006 [cited by applicant]
US 20060251576A1 · Hellerstein · 2006 [cited by applicant]
US 20080279766A1 · Everson et al. · 2008 [cited by applicant]
US 20100055734A1 · Everson · 2010 [cited by applicant]
US 20120329161A1 · Everson et al. · 2012 [cited by applicant]
US 20140067276A1 · Everson et al. · 2014 [cited by applicant]
US 20140147875A1 · Everson et al. · 2014 [cited by applicant]
US 20140326926A1 · Everson et al. · 2014 [cited by applicant]
US 20150204842A1 · Everson et al. · 2015 [cited by applicant]
US 20150268255A1 · Everson et al. · 2015 [cited by applicant]
US 20160305930A1 · Everson et al. · 2016 [cited by applicant]
AU 2012262329 · 2014 [cited by applicant]
EP 1150123 · 2001 [cited by applicant]
JP 2003532899 · 2003 [cited by applicant]
JP 2004507740 · 2004 [cited by applicant]
JP 2010502979 · 2010 [cited by applicant]
WO 216949 · 2002 [cited by applicant]
WO 2008029145 · 2008 [cited by applicant]
WO 2012166802 · 2012 [cited by applicant]
WO 2014075082 · 2014 [cited by applicant]
Zimmermann et al., PLoS One, 2011, 6(6) e21381, pp. 1-10. [cited by examiner]
I.R. Mackay. Chronic Hepatitis: Effect of Prolonged Suppressive Treatment and Comparison of Azathioprine With Prednisolone. Q J Med, 1968, 37(147):379-392. (Year: 1968). [cited by examiner]
Wallack et al. Non-Invasive Measurement of the Portal Circulation Using Cholates Quantifies Disease Severity in Primary Sclerosing Cholangitis Gas. 2012, 142: S911. (Year: 2012). [cited by examiner]
Zimmermann et al. Interleukin-8 Is Activated in Patients with Chronic Liver Diseases and Associated with Hepatic Macrophage Accumulation in Human Liver Fibrosis. PLoS One, 2011, 6(6) e21381, pp. 1-10. (Year: 2011). [cited by examiner]
Afdhal et al., “Advances in Hepatology—Fibroscan (Transient Elastography) for the Measurement of Liver Fibrosis”; Sec. Ed. Schiff, Gastroenterology & Hepatology; 8(9):605-607 (Sep. 2012). [cited by applicant]
Bio Predictive, “Technical Recommendations for FibroTest and FibroMax assays”; A Guide for biologists and laboratories, V. 1.17, 28 pgs (Oct. 20, 2014). [cited by applicant]
Chronic Hepatitis Data Sheet, Merck, Sharp & Dohme, Corp., 1 page (2010-2011). [cited by applicant]
Dax et al., “HPLC-Continuous-Flow Fast Atom Bombardment Mass Spectrometry (HPLC-CFFAB)—a Convenient Method for the Analysis of Bile Acids in Bile and Serum”; Chromatographia, 40(11/12):674-679 (Jun. 1995). [cited by applicant]
Decompensated Cirrhosis Data Sheet, U.S. Department of Veterans Affairs, 1 page (2011). [cited by applicant]
DeMark, “A method for the accurate measurement of isotope ratios of chenodeoxycholic and cholic acids in serum”; J Lipid Res, 23:204-210 (1982). [cited by applicant]
Denaro et al., “The effect of liver disease on urine caffeine metabolite ratios”; Clinical Pharmacology and Therapeutics, 59:624-635 (1996). [cited by applicant]
Di Bisceglie, “Prolonged therapy of advanced chronic hepatitis C with low-dose peginterferon”; N Engl J Med, 359 (23):2429-2441 (Dec. 4, 2008). [cited by applicant]
Eichelbaum, “Simultaneous Determination of the intravenous and Oral Pharmacokinetic Parameters of D,L-Verapamil Using Stable Isotope-Labelled Verapamil”; Eur J Clin Pharmacol, 19:133-137 (1981). [cited by applicant]
European Search Report for Application No. 10815965.8 mailed Apr. 17, 2013. [cited by applicant]
Everson et al., “Quantitative Tests (QLFTS) Detect Impaired Hepatic Function in a High Proportion of Chronic Hepatitis C Patients With Fibrosis or Compensated Cirrhosis and may Predict Risk of Cirrhosis, Splenomegaly, a… [cited by applicant]
Everson et al., “Qualitative Tests (QLFTS) Detect Impaired Hepatic Function in a High Proportion of Patients with Chronic HCV and Firbrosis or Cirrhosis and May Predict Risk of Cirrhosis, Splenomegaly and Varices”, pres… [cited by applicant]
Everson et al., “Portal-systemic shunting in patients with fibrosis or cirrhosis due to chronic hepatitis C: the minimal model for measuring cholate clearances and shunt”, Alimentary Pharmacology & Therapeutics, 26:401-… [cited by applicant]
Everson et al., “The spectrum of hepatic functional impairment in compensated chronic hepatitis C: results from the Hepatitis C Anti-viral Long-term Treatment against Cirrhosis Trial”; Alimentary Pharmacology & Therapeu… [cited by applicant]
Everson et al., “Hepatic Impairment Measured by Quantitative Tests of Liver Function (QLFTs) Predicts Clinical Outcome in Patients with Advanced Fibrosis: Results from the Hepatitis C Anti-viral Long-term Treatment agai… [cited by applicant]
Everson et al. “Quantitative tests of liver function measure hepatic improvement after sustained virological response: results from the HALT-C trial”; Alimentary Pharmacology & Therapeutics, 29:589-601(2009). [cited by applicant]
Everson et al., “Quantitative Liver Function Tests Improve the Prediction of Clinical Outcomes in Chronic Hepatitis C: Results from the Hepatitis C Antiviral Long-term Treatment Against Cirrhosis Trial”; Hepatology, 55(… [cited by applicant]
Everson et al., “Functional Elements Associated with Hepatic Regeneration in Living Donors After Right Hepatic Lobectomy,” Liver Transplantation, vol. 19, No. pp. 292-304 (2013). [cited by applicant]
Exalenz Breathid® breath test device for the diagnosis of liver disease; Health Policy Advisory Committee on Technology—Technology Brief (Aug. 2012). [cited by applicant]
Examination Report for Australian Application No. 2013341378, mailed Jan. 17, 2017, 3 pages total. [cited by applicant]
Examination Report No. 1 from related Australian patent application 2012262329, mailed Jun. 23, 2014. [cited by applicant]
Extended European Search Report for EP 06734026.5, mailed Mar. 31, 2011. [cited by applicant]
Gilmore et al., “Plasma clearance of oral and intravenous cholic acid in subjects with and without chronic liver disease”; Gut, 21:123-127 (1980). [cited by applicant]
Golden et al., “Application of an enzyme-multiplied immunoassay technique for determination of caffeine elimination kinetics as a test of liver function in clinically normal dogs”; American Journal of Veterinary Researc… [cited by applicant]
Guidance for Industry, Bioanalytical Method Validation, (May 2001). [cited by applicant]
Haque et al., “Hepatitis C antiviral long-term treatment against cirrhosis (HALT-C) trial”; Annals of Hepatology, 8 (1):78-79 (Jan.-Mar. 2009). [cited by applicant]
HCV Fibrosure™, Informational Sheet; Laboratory Corporation of America (2004). [cited by applicant]
Hechey et al., “Syntheses with stable isotopes: synthesis of deuterium and 13C labeled bile acids,” Journal of Labelled Compounds, IX(4):703-719 (Oct.-Dec. 1973). [cited by applicant]
Helmke et al., “Slow, Moderate, and Rapid Progressors: Three Distinct Categories of Patients with Primary Sclerosing Cholangitis Detected by Functional Assessment using Cholate Testing,” Hepatology, 56(4)(Suppl):1133A, … [cited by applicant]
Helmke et al., “Noninvasive assessment of liver function”; Liver—Current Opinion in Gastroenterology 31(3):1-10 (Feb. 31, 2015). [cited by applicant]
Herold et al., “Quantitative testing of liver function in patients with cirrhosis due to chronic hepatitis C to assess disease severity”; Liver, 21:26-30 (2001). [cited by applicant]
Hoofnagle, “Course and outcome of hepatitis C”; Hepatology, 36(5):S21 (Nov. 2002). [cited by applicant]
Hydzik et al., “Usefulness of 13C-methacetin breath test in liver function testing in Amanita phalloides poisoning: breast feeding woman case”; Clinical Toxicology, 46:1077-1082 (2008). [cited by applicant]
International Search Report for PCT/US06/03132, mailed Jul. 11, 2007. [cited by applicant]
International Search Report and Written Opinion for PCT/US10/47976, mailed Feb. 2, 2011. [cited by applicant]
International Search Report and Written Opinion cited in PCT/US2012/040008 mailed Sep. 6, 2012. [cited by applicant]
International Search Report and Written Opinion for PCT/US13/69708 mailed Feb. 10, 2014. [cited by applicant]
Invitation to Pay Additional Fees for PCT/US10/47676, dated Nov. 17, 2010. [cited by applicant]
Japanese Office Action mailed Aug. 28, 2017 in Japanese patent application No. 2015-542017 and English language translation, 8 pages total. [cited by applicant]
Kamath et al., “A Model to Predict Survival in Patients with End-Stage Liver Disease”; Hepatology, 33(2):464-470 (Feb. 2001). [cited by applicant]
Kern, “Normal Plasma Cholesterol in an 88-year-old Man Who Eats 25 Eggs a Day”; New England J Medicine, 324:896-899 (1991). [cited by applicant]
Koster et al., “Recent Developments in On-line SPE for HPLC and LC-MS in Bioanalysis”; Guide to LC-MS, 3 pp. (Dec. 2001). [cited by applicant]
Krumbiegel et al., “[15N]methacetin urine test: a method to study the development of hepatic detoxification capacity”; Eur J Pediatr., 149(6):393-395 (Mar. 1990). [cited by applicant]
Lalazar et al., “A continuous 13C methacetin breath test for noninvasive assessment of intrahepatic inflammation and fibrosis in patients with chronic HCV infection and normal ALT”; Journal of Viral Hepatitis 15(10):716… [cited by applicant]
Mackay, Q J Med., 1968, 37(147):379-392. [cited by applicant]
Martucci, “Deconvolutional Analysis on Clearance Curves of Simultaneously Administered Oral and Intravenous Doses of 2,2,4,4-2H Cholate and 24-13C Cholate: Minimal Model to Determine First-Pass Hepatic Extraction of Cho… [cited by applicant]
Medrzejewski et al., “Plasma Clearance of Cholic Acid in Patients With Chronic Diseases of the Liver”; Polski Tygodnik Lekarski, 45(16-18):335-337, Abstract Only, 1 page. (Apr. 16-30, 1990). [cited by applicant]
Miescher et al., “Portal-systemic spill-over of bile acids: a study of mechanisms using ursodeoxycholic acid”; European J of Clinical Investigation; 13:439-445 (1983). [cited by applicant]
Nguyen et al., “Diagnostic and Therapeutic Advances in Hepatology-Noninvasive Assessment of Liver Fibrosis”; Hepatology; 2107-2110 (Jun. 2011). [cited by applicant]
Office Action for EP 12731209.8 dated Sep. 7, 2016, 4 pages total. [cited by applicant]
Queiroz et al., “Practical Tips on Preparing Plasma Samples for Drug Analysis Using SPME,” LCGC North America, vol. 22, No. 10, 6 pages (Oct. 2004). [cited by applicant]
Ratziu et al., “Diagnostic value of biochemical markers (Fibro Test-FibroSURE) for the prediction of liver fibrosis in patients with non-alcoholic fatty liver disease”; BMC Gastroenterology; 6:6 (Feb. 14, 2006). [cited by applicant]
Rector et al., Renal sodium retention complicating alcoholic liver disease: Relation to portosystemic shunting and liver function Hepatology 12(3):455-459 (1990). [cited by applicant]
Renner et al., “Caffeine: A Model Compound for Measuring Liver Function”, Hepatology 4(1):38-46 (1984). [cited by applicant]
Shah et al., “Comparison of Noninvasive Markers of Fibrosis in Patients With Nonalcoholic Fatty Liver Disease”; Clinical Gastroenterology and Hepatology; 7(10):1104-1112 (2009). [cited by applicant]
Shrestha et al., “Quantitative liver function tests define the functional severity of liver disease in early-state cirrhosis”; Liver Transplantation and Surgery 3(2):166-167, 172 (Mar. 1997). [cited by applicant]
Stellaard, et al., “Measurement of Bile Acid Kinetics in Human Serum Using Stable Isotope Labeled Chenodeoxycholic Acid and Capillary Gas Chromatograpy Electron Impact Mass Spectrometry,” Biomedical Mass Spectrometry, 1… [cited by applicant]
Stellaard, et al., “Simultaneous determination of cholic acid and chenodeoxycholic acid pool size and fractional turnover rates in human serum using 13C-labeled bile acids”; Journal of Lipid Research, 25:1313-1319 (1984… [cited by applicant]
Stellaard, “Simultaneous determination of pool sizes and fractional turnover rates, of deoxycholic acid, cholic acid and chenodeoxycholic acid in man by isotope dilution with 2H and 13C labels and serum sampling,” Biome… [cited by applicant]
Supplementary European Search Report for Application No. 13853943.2 mailed May 23, 2016, 8 pages total. [cited by applicant]
Toyoda et al., Clin Gas and Hepatology, 2006, 4:1528-1536. [cited by applicant]
Wallack et al., “Non-invasive measurement of the portal circulation using cholates quantifies disease severity in primary sclerosing cholangitis”, Gastroenterology, vol. 142, No. 5, suppl. 1, p. S911 (May 1, 2012). [cited by applicant]