A new cirrhotic animal protocol combining carbon tetrachloride with methotrexate to address limitations of the currently used chemical-induced models.

Background: Chemically induced cirrhotic animal models are commonly used. However, they have limitations such as high mortalities and low yield of cirrhotic animals that limit their uses. Aims: To overcome limitations of the chemically induced cirrhotic animal model via combined administration of methotrexate (MTX) with CCl4 and decrease their commonly used doses depending on the proposed synergetic cirrhotic effect. Methods: Rats were divided into six groups: normal (4 weeks), normal (8 weeks), MTX, CCl4 (4 weeks), CCl4 (8 weeks), and MTX + CCl4 (4 weeks) groups. Animals' hepatic morphology and histopathological characterization were explored. Hepatic Bcl2 and NF-κB-p65 tissue contents were determined using the immunostaining technique, and hepatic tissue damage, oxidative status, and inflammatory status biochemical parameters were determined. Results: CCl4 + MTX combined administration produced prominent cirrhotic liver changes, further confirmed by a substantial increase in oxidative stress and inflammatory parameters, whereas mortalities were significantly lower than in other treated groups. Conclusion: The present study introduced a new model that can significantly improve the major limitations of chemically induced cirrhotic animal models with new pathological features that mimic human cirrhosis. Compared to other chemically induced methods, the present model can save time, cost, and animal suffering.

Analytical Purity Determinations of Universal Food-Spice Curcuma longa through a QbD Validated HPLC Approach with Critical Parametric Predictors and Operable-Design's Monte Carlo Simulations: Analysis of Extracts, Forced-Degradants, and Capsules and Tablets-Based Pharmaceutical Dosage Forms.

Applications of analytical quality by design (QbD) approach for developing HPLC (High Performance Liquid Chromatography) methods for food components assays, and separations of complex natural product mixtures, are still limited. The current study developed and validated, for the first time, a stability-indicating HPLC method for simultaneous determinations of curcuminoids in Curcuma longa extracts, tablets, capsules, and curcuminoids' forced degradants under different experimental conditions. Towards separation strategy, critical method parameters (CMPs) were defined as the mobile phase solvents' percent-ratio, the pH of the mobile phase, and the stationary-phase column temperature, while the peaks resolution, retention time, and the number of theoretical plates were recognized as the critical method attributes (CMAs). Factorial experimental designs were used for method development, validation, and robustness evaluation of the procedure. The Monte Carlo simulation evaluated the developing method's operability, and that ensured the concurrent detections of curcuminoids in natural extracts, commercial-grade pharmaceutical dosage-forms, and the forced degradants of the curcuminoids in a single mixture. The optimum separations were accomplished using the mobile phase, consisting of an acetonitrile-phosphate buffer (54:46 v/v, 0.1 mM) with 1.0 mL/min flow rate, 33 °C column temperature, and 385 nm wavelength for UV (Ultra Violet) spectral detections. The method is specific, linear (R2 ≥ 0.999), precise (% RSD < 1.67%), and accurate (% recovery 98.76-99.89%), with LOD (Limit of Detection) and LOQ (Limit of Quantitation) at 0.024 and 0.075 µg/mL for the curcumin, 0.0105 µg/mL and 0.319 µg/mL for demethoxycurcumin, and 0.335 µg/mL and 1.015 µg/mL for the bisdemethoxycurcumin, respectively. The method is compatible, robust, precise, reproducible, and accurately quantifies the composition of the analyte mixture. It exemplifies the use of the QbD approach in acquiring design details for developing an improved analytical detection and quantification method.

Determination of serum fibrosis index in patients with chronic hepatitis and its relationship to histological activity index.

OBJECTIVE: To study the reliability of serum hyaluronic acid (HA) as a non-invasive method for the diagnosis of liver fibrosis and its relationship to liver biopsy findings. METHODS: In a prospective controlled clinical trial, 48 patients with chronic liver disease were selected from Pediatric Departments, Al-Jedaany and Al-Hayat Hospitals, Jeddah, Kingdom of Saudi Arabia, from November 2005 to March 2008. Twenty-one with chronic hepatitis B infection, 17 with chronic hepatitis C infection, and 10 with autoimmune hepatitis in addition to 25 healthy controls. Serum HA and liver function tests were carried out for the studied cases. The value of HA was correlated with the histopathologic findings of liver biopsy in chronic hepatitis patients. RESULTS: Serum HA increased significantly in chronic hepatitis cases compared with control. The mean serum HA was 111.22 mg/L for patients with chronic hepatitis B, 113.05 mg/L for hepatitis C, 112.30 mg/L for autoimmune hepatitis, and 33.96 mg/L for control group. Serum HA significantly increased in chronic hepatitis patients with stage 2 (p=0.0029) and stage 3 (p=0.0013) fibrosis compared with stage 0 (p=0.0054) fibrosis. Serum HA positively correlated with the degree of liver fibrosis, it increased significantly with stage 3 fibrosis (157.96 mg/L) compared with stage 2 fibrosis (122.13 mg/L) (p=0.0013). CONCLUSION: Serum HA increased in chronic hepatitis, and its level correlates with the degree of fibrosis. Serum HA levels can be used for diagnosis and followed up of liver fibrosis in patients with chronic hepatitis.