R. Naccarato

Iron storage, lipid peroxidation and glutathione turnover in chronic anti-HCV positive hepatitis

BACKGROUND/AIMS Little is known about the pathogenesis of liver damage related to hepatitis C virus. The presence of steatosis or increased ferritin levels, and preliminary data on the relevance of iron as a prognostic factor prompted us to ascertain whether hepatitis C virus-related liver damage might be mediated by iron accumulation. METHODS We evaluated the degree of hepatic inflammation and steatosis, serum ferritin, transferrin saturation and iron levels, tissue iron concentrations and iron index, liver glutathione and malondialdehyde in 33 males and 20 females with chronic hepatitis C virus- or hepatitis B virus-related hepatitis (42 + 11). We also considered six patients with both alcohol abuse and hepatitis C virus, four males with chronic alcoholic liver disease and four males with genetic hemochromatosis, giving a total of 67. All diagnoses were histologically confirmed. Patients with cirrhosis were excluded. RESULTS Our data show that: 1. Steatosis is more frequent in hepatitis C virus and hepatitis C virus+alcohol abuse patients; 2. In males, serum ferritin and tissue iron are significantly higher in hepatitis C virus- than in hepatitis B virus-positive patients (p < 0.01 and 0.05); transferrin saturation is higher (p < 0.05) in hepatitis C virus-positive than in hepatitis B virus-positive patients only when males and females are considered together; 3. Serum ferritin and transferrin saturation only correlate with liver iron (r = 0.833 and r = 0.695, respectively, p = 0.00001); tissue iron is significantly higher in hepatitis C virus- than in hepatitis B virus-positive patients (p < 0.05); 4. In patients with chronic hepatitis, serum ferritin is a better marker of liver iron storage than transferrin saturation, both in males and in females; 5. Hepatitis C virus-positive patients have higher malondialdehyde levels and activation of turnover of glutathione, probably in response to free-radical-mediated liver damage. Females have lower liver iron levels but similar trends. CONCLUSIONS These findings suggest that hepatitis C virus-related liver damage is characterized by increased iron storage (possibly induced by the virus) which elicits a free-radical-mediated peroxidation, with consequent steatosis and activation of glutathione turnover.

Oxidative DNA damage accumulation in gastric carcinogenesis

Background—Gastric carcinogenesis is a multifactorial, multistep process, in which chronic inflammation plays a major role. Aims—In order to ascertain whether free radical mediated oxidative DNA damage is involved in such a process, concentrations of 8-hydroxydeoxyguanosine (8OHdG), a mutagenic/carcinogenic adduct, and thiobarbituric acid reactive substances (TBARS), as an indirect measure of free radical mediated damage, were determined in biopsy specimens from patients undergoing endoscopy. Patients—Eighty eight patients were divided into histological subgroups as follows: 27 with chronic non-atrophic gastritis, 41 with atrophic gastritis, six with gastric cancer, and 14 unaffected controls. Methods—Intestinal metaplasia,Helicobacter pylori infection, and disease activity were semiquantitatively scored. 8OHdG concentrations were assessed by HPLC with electrochemical detection, and TBARS concentrations were fluorimetrically assayed. Results—8OHdG concentrations (mean number of adducts/105 dG residues) were significantly higher in chronic atrophic gastritis (p=0.0009). Significantly higher concentrations were also detected in the presence of severe disease activity (p=0.02), intestinal metaplasia (p=0.035), and H pylori infection (p=0.001). TBARS concentrations were also higher in atrophic gastritis, though not significantly so. In a multiple logistic regression analysis, 8OHdG concentrations correlated best with the presence and severity of H pylori infection (r=0.53, p=0.002). Conclusions—Chronic gastritis is characterised by the accumulation of oxidative DNA damage with mutagenic and carcinogenic potential. H pylori infection is the major determinant for DNA adduct formation.

How should patients with hepatocellular carcinoma be staged? Validation of a new prognostic system

The life expectancy of a patient with hepatocellular carcinoma (HCC) in cirrhosis is hard to predict, making it difficult to decide whether a certain treatment is indicated and what to say to the patient regarding prognosis. A new score recently has been proposed, which includes the parameters involved in the Child‐Pugh stage, plus macroscopic tumor morphology, α‐fetoprotein levels, and the presence or absence of portal thrombosis. The score has been validated in internal control series, but its general applicability has yet to be confirmed. The authors compared the discriminatory ability of the Cancer of the Liver Italian Program (CLIP) score with those of the Okuda and TNM staging systems and the Child‐Pugh classification in a group of cirrhotic patients with HCC, diagnosed and followed up by their unit.

Oxidative DNA damage in circulating leukocytes occurs as an early event in chronic HCV infection.

UNLABELLED Chronic hepatitis C virus (HCV) infection is associated with an increased production of reactive oxygen species within the liver that are responsible for the oxidation of intracellular macromolecules. To ascertain whether the increased risk of hepatocellular carcinoma in individuals with chronic HCV infection is related to an accumulation of oxidative DNA damage, the 8-hydroxydeoxyguanosine (8-OHdG) content in the DNA of liver tissue and leukocytes of 87 individuals with HCV- or HBV-related liver disease and of 10 healthy controls was measured. Serum levels of thiobarbituric acid reactive substances (TBARS) were also assessed as an index of lipid peroxidation. RESULTS The 8-OHdG content in the circulating leukocytes correlated with that of liver tissue (r = 0.618, p < .0004). HCV patients had the highest median 8-OHdG levels (p < .0004). 8-OHdG leukocyte levels in HCV patients were higher than in HBV patients (p < .04) and they significantly correlated with the clinical diagnosis (p < .025), the serum ferritin levels (p < .05), and the amount of liver steatosis (p < .001). No correlation was found with age, gender, history of drinking or smoking, ALT or GGT levels, ESR, alpha-1, or gamma-globulin level and Ishak score. TBARS levels were significantly higher in cirrhotics than in noncirrhotics (p < .01). CONCLUSIONS The 8-OHdG level in circulating leukocytes is a reliable marker of oxidative stress occurring in the liver of individuals with chronic HCV infection. DNA oxidative damage appears to be an early and unique event in the natural history of HCV-related hepatitis. This injury increases the risk of genomic damage and may be one of the important factors involved in the carcinogenic process in cases of HCV-related chronic liver disease.

Risk factors in community-acquired chronic hepatitis C virus infection: a case-control study in Italy.

AIMS/METHODS A case-control study was carried out in Italy to assess the risk factors associated with chronic hepatitis C virus infection. Five hundred consecutive chronic anti-hepatitis C virus positive cases and 500 sex and exactly age-matched anti-hepatitis C virus negative/HBsAg negative controls entered the study. Information was collected through an interviewer-administered questionnaire. The adjusted Odds Ratios linking hepatitis C virus infection and risk factors were estimated by conditional multiple logistic regression. Demographic and socio-economic characteristics were similar in cases and controls. Seventy-five percent of patients were aged over 40: males were prominent in the group < or = 40, while the number of females increased with age. RESULTS As expected, drug addiction and blood transfusion emerged as independent risk factors: blood transfusion in all ages and in both sexes, drug addiction only in subjects under 41 years and mostly in males. Other risk factors independently associated with hepatitis C virus infection were: previous use of non-disposable needles, previous tuberculosis, and prolonged hospitalization before 1970. A history of sexually-transmitted diseases was not associated. CONCLUSIONS This study shows that the great spread of hepatitis C virus in Italy may have occurred several years ago through parenteral routes which are not now operating. Modern hygienic and sanitation measures have significantly controlled exposure to the infection, which in the younger generations is confined to high-risk groups such as drug addicts.

Discrepancies Between Reported Food Intolerance and Sensitization Test Findings in Irritable Bowel Syndrome Patients

OBJECTIVES:Irritable bowel syndrome (IBS) is a common gastrointestinal disorder with clinical signs typical of “intestinal” food allergies or intolerance. The aim of this study was to characterize the clinical features of IBS patients suspected of suffering from adverse reactions to food.METHODS:The study involved 128 consecutive IBS patients divided into four groups according to their main symptom on presentation at our outpatient clinic. A detailed medical history was recorded, paying particular attention to any allergies and reported intolerance to food. Each patient was screened for allergies; intestinal permeability tests was performed in randomly selected patients from different groups. Findings were analyzed using the χ2 test.RESULTS:Adverse reactions to one or more foods were reported by 80 patients (62.5%); skin prick tests (SPT) were positive in 67 patients (52.3%) with no significant differences between patients complaining of different symptoms. Patients who reported a food intolerance had more positive SPTs than those who did not (47 of 80 [58.7%] vs 20 of 48 [41.7%]); this difference was not statistically significant, although it suggests a trend (p < 0.0610). There was little consistency between the specific foods reported to cause intolerance and those resulting from the tests (11 of 80 patients, 13.7%). The intestinal permeability test was normal in 29 of 33 patients (87.9%).CONCLUSIONS:More than 50% of IBS patients were found sensitized to some food or inhalant without any typical clinical signs. Patients were unable to identify potentially offending foods. The lack of a correlation between SPT results and reported food allergies needs further investigation to clarify the pathophysiology and improve the diagnosis of intestinal food allergies.

Evolution of osteopenia in inflammatory bowel disease

Objective:Our aim was the assessment of frequency and evolution of osteopenia in patients with inflammatory bowel disease and identification of related factors.Methods:Bone mineral density (BMD) of the lumbar spine was measured in 54 patients with Crohns disease (CD) and in 49 patients with ulcerative colitis (UC) and was repeated after a mean observation period of 21 (range, 8–50) months in 30 CD and 14 UC patients. Eighteen age-matched healthy subjects served as controls. Serum biochemistry (parathyroid hormone, osteocalcin, alkaline phosphatase, insulin-like growth factor 1, minerals, and markers of inflammation) was assessed at the time of the second BMD measurement.Results:Reduced BMD values were found in 48% of CD, and in 38% of UC patients. Compared with control subjects, the mean BMD was significantly lower in CD p < 0.003 and UC p < 0.0001 patients. BMD was positively correlated with the body mass index p < 0.05 and inversely correlated with the lifetime steroid dose p < 0.03. After 21 months the BMD of CD patients was virtually unchanged, with an annual variation (%ΔBMD/yr) of −0.31 ± 0.49, whether treated with steroids or not, whereas in UC patients the BMD decreased significantly p < 0.02 with a %ΔBMD/yr of −2.47 ± 0.82 (p < 0.02vs CD). This decrease can be attributed to steroid treatment. No biochemical alterations were detected in patients with rapid bone loss, compared with those with stable BMD.Conclusions:Low bone density is frequent in both CD and UC, but apparently stable in CD. The evolution of BMD suggests that low bone density is associated with the pathogenesis of CD, whereas in UC it seems to be correlated with the side effects of corticosteroid treatment.

The impact of liver disease and medical complications on quality of life and psychological distress before and after liver transplantation.

BACKGROUND/AIM The impact of liver disease and medical complications on quality of life (QOL) and psychological distress before and after orthotopic liver transplantation (OLT) is a matter of growing interest. METHODS Perceived QOL (LEIPAD Quality of Life test) and psychological distress (Brief Symptom Inventory, BSI) were assessed in 40 cirrhotic patients listed for OLT (Group A) and in 101 liver transplant recipients (Groups B to G=0-6, 7-12, 13-24, 25-36, 37-48, 49-60 months post-OLT). Patients were also evaluated for medical complications, blood levels of immunosuppressive agents and recurrence of liver disease. RESULTS QOL and psychological distress were significantly better in most of the post-OLT groups than in cirrhotic patients. Among post-OLT patients, a significantly worse QOL was perceived at 13-24 months (Life Satisfaction: Group D vs G, p=0.024; Cognitive Functioning: Group D vs F, p=0.024), while significantly greater psychological distress was detected at 7-12 months (Anxiety and Interpersonal Sensitivity: Group C vs Group B, p=0.032 and p=0.023, respectively). Medical complications and immunosuppressive therapy did not influence QOL or psychological distress after OLT. Within 6 months after OLT, patients with HCV recurrence showed significantly greater Depression (p=0.023), Anxiety (p=0.038), Phobic Anxiety (p=0.001), and Paranoid Ideation (p=0.033) than anti-HCV negative patients. CONCLUSIONS Liver transplantation improves psychological distress and most, but not all, QOL domains. Recurrent HCV infection is associated with greater psychological distress.

The role of cysteine and serine proteases in colorectal carcinoma

Cathepsin B (CATB) and cathepsin L (CATL), which are cysteine proteases, urokinase‐(UPA) and tissue‐type plasminogen activator (TPA), both serine proteases, and their inhibitor type‐1 (PAI‐1) are believed to play an important role in colorectal carcinoma (CRC) invasion and metastasis. The objective of this study was to measure CATB, CATL, UPA, TPA, and PAI‐1 in the same cancerous tissue (CANCER) and in tissues obtained from a tumor free area (NORMAL) to compare their respective prognostic roles in patients with CRC.

Altered esophageal pain threshold in irritable bowel syndrome

Gut motility disorders and altered pain perception were reported in patients with irritable bowel syndrome (IBS). To verify foregut involvement in IBS, we studied 30 patients using esophageal manometry and 24-hr pH monitoring of the distal esophagus. Two subgroups of patients underwent esophageal provocative tests (bethanechol 50 μg/kg subcutaneously and esophageal balloon distension test). Twelve healthy volunteers formed a control group. A pain threshold on esophageal distension significantly lower than in healthy subjects (11.5±1 ml vs 22.2±1.7 ml,P<0.01) was found in IBS patients. On the other hand, no differences between patients and controls were detected in lower esophageal sphincter pressure and length, esophageal body motility, or GER pattern; furthermore, bethanechol stimulation elicited similar esophageal body motility changes. Our study could confirm no detectable basal or bethanechol-induced esophageal motility disorders in IBS patients, nor enhanced GER. Esophageal involvement in IBS consists of a lower pain threshold on esophageal distension, possibly reflecting an altered visceral receptor sensitivity or modulation throughout the gut.