In a one year period we studied 98 liver patients consecutively admitted to the III Department of Internal Medicine, Federico II University, Naples, Italy. Among them we selected 25 patients affected by biopsy-proven chronic hepatitis or liver cirrhosis, virus-related, according to the following inclusion criteria: male sex, age < 60 years, body mass index (BMI) from 20 to 30 kg/m2, normal physical activity, biopsy-proven liver cirrhosis only in Child’s A class, virus-related.
All patients lived in the same geographical area from Southern Italy. Their diet was a tipical mixed, mediterranean diet, containing milk and cheese products, without alcohol intake; only five patients consumed less than 20 g of alcohol three or four times a week. Information about dietary habits and alcohol intake were carefully obtained from a food frequency questionnaire.
Exclusion criteria were: female sex, age > 60 years, alcohol abuse, drug addiction, obesity, previous bone fractures or history of recent immobilization, autoimmune, endocrinological or rheumatological diseases, treatment with corticosteroids, diuretics, drugs affecting bone metabolism or interferon for more than three months over the past three years. Advanced liver cirrhosis (B or C Child’s class), hepatocellular carcinoma, primary biliary cirrhosis, cholestatic liver diseases, genetic he- mochromatosis were also considered exclusion criteria. The selected patients had positive serological markers for viral hepatitis (2 for hepatitis B virus and 23 for hepatitis C virus) and biopsy-proven diagnosis of chronic liver disease: 12 patients were affected by chronic hepatitis and 13 patients by liver cirrhosis, A Child’s class. Liver function was well preserved and renal function was normal in all patients. None exhibited indices of cholestasis nor experienced previous decompensation of cirrhosis.
Thirthy healthy male volunteers, living in the same geographical area, age, and BMI-matched, were also examined as control group.
All patients and controls received information about the study and gave their informed consent to participate. Bone mineral density (BMD) was measured at lumbar spine (L1-L4) and at proximal femur of non dominant leg by Dual-Energy X-ray Absorptiometry (DEXA) using a Hologic QDR 1000 densitometer.
Osteoporosis was defined in analogy with the World Health Organization criteria for women. Individual BMD values are expressed as gr/cm2 and T- and Z-score. Quality control was maintained by daily scanning of an anthropomorphic spine phantom. The coefficient of variation for the DEXA technique was less than 1% for the lumbar spine (LS) and 1.5% for the femoral neck (FN). The reference population adopted in this study was the international pooled sample provided by the manifacturer; their data, however, did not differ significantly for those obtained on a local sample in a study performed when the device was set up.
Biochemical tests of liver function and bone metabolism in patients and controls were carried out after an overnight fast. Biochemical liver function tests and serum calcium, inorganic phosphate, and magnesium were measured with an automatic technique. Serum was assayed using commercially available kits for intact Parathyroid hormone (PTH), (IRMA, Incstar Corporation), 25-hydroxyvitamin D (25-OHD), (RIA Incstar Corporation), IGF-1, (RIA, Nichols Institute, San Juan Capistrano, CA, USA), bone isoenzyme alkaline phosphatase (b-Ap) as a marker of bone formation (Tandem-T, Ostase, IRMA, Hybritech Europe, Liege, Belgium), Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH), (RIA, Biodata, Serono Diagnostics, Italy), Prolactin (HPRL), Oestrogen, Progesterone, 17-OH- Progesterone, Delta-4-Androstenedione, Testosterone (RIA, Diagnostics Products Corporation, Los Angeles, CA, USA), Adrenocorticotropic hormone (ACTH), Cortisol, Aldosterone, complete Thyroid Hormones and antibodies profile (FT3, FT4, T3, T4, TSH, Tg, TgAb, TPOAb), (RIA, Sorin Biomedica, Saluggia, Italy). You can shop for the medicine you need for treating the condition mentioned above right here at the Canadian Health&Care Mall. This pharmacy offers a large selection of medications available over the internet at amazingly low prices.
Urine was analyzed for calcium (automatic absorption spec- trophotometry) and creatinine (standard automated techniques). Urinary hydroxyproline, as a marker of bone resorption, was measured with a commercial Kit (OHP Biolab, Italy) and urinary excretion was expressed in terms of urinary creati- nine as the ratio of hydroxyproline to creatinine. Statistical analysis: results were expressed as means ± SD. Student’s unpaired t-test was used to compare means between the groups. The non parametric Mann-Whitney U test was used when Wik-Shapiro test was not consistent with the Gaussian distribution of the data. Linear multiple regression analysis (using as covariates age and BMI) was used to evaluate the determinants of bone mass. Significance was retained for p < 0.05.