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There is increasing evidence that reactive oxygen species play a pivotal role in the process of ageing. The skin, as the outermost barrier of the body, is exposed to various exogenous sources of oxidative stress, in particular UV-irradiation. These are believed to be responsible for the extrinsic type of skin ageing, termed photo-ageing. It therefore seems reasonable to try to increase levels of protective low molecular weight antioxidants through a diet rich in fruits and vegetables or by direct topical application. Indeed, various in vitro and animal studies have proved that low molecular weight antioxidants, especially vitamins C and E, ascorbate and tocopherol, as well as lipoic acid, exert protective effects against oxidative stress. However, controlled long-term studies on the efficacy of low molecular weight antioxidants in the prevention or treatment of skin ageing in humans are still lacking.

Background: Combination therapy using ursodeoxycholic acid plus chenodeoxycholic acid has been advocated for dissolution of cholesterol gallstones because the two bile acids have complementary effects on biliary lipid metabolism and cholesterol solubilization. Aim: To compare the clinical efficacy of combination therapy with ursodeoxycholic acid monotherapy. Patients and methods: A total of 154 symptomatic patients with radiolucent stones (≤ 15 mm) in functioning gallbladders were enrolled from six centres in England and Italy. They were randomized to either a combination of chenodeoxycholic acid plus ursodeoxycholic acid (5 mg.day/kg each) or to ursodeoxycholic acid alone (10 mg.day/kg). Dissolution was assessed by 6-monthly oral cholecystography and ultrasonography for up to 24 months. Results: Both regimens reduced the frequency of biliary pain and there was no significant difference between them in terms of side-effects or dropout rate. Complete gallstone dissolution on an intention-to-treat basis was similar at all time intervals. At 24 months this was 28% with ursodeoxycholic acid alone and 30% with combination therapy. The mean dissolution rates at 6 and 12 months were 47% and 59% with ursodeoxycholic acid, and 44% and 59% with combination therapy, respectively. Conclusion: There is no substantial difference in the efficacy of combined ursodeoxycholic acid and chenodeoxycholic acid and that of ursodeoxycholic acid alone in terms of gallstone dissolution rate, complete gallstone dissolution, or relief of biliary pain.

SUMMARY The autoimmune nature of primary biliary cirrhosis (PBC) is well established. We tested the hypothesis that fetal microchimerism indicated by the persistence of circulating fetal cells in women years after pregnancy might contribute to the aetiopathogenesis of PBC through a graft-versus-host-like response. We extracted DNA from the peripheral blood cells of 36 women carefully selected from 173 consecutive PBC patients, who were matched with 36 healthy women by age, age of last son, and number of children. Both patients and controls had to have male offspring, and no history of miscarriages or blood transfusions; they could not be twins. We tested all of the samples for the presence of two specific Y-chromosome sequences (SY154 and SRY) by amplifying DNA in a nested polymerase chain reaction. Y-chromosome-specific DNA was detected in the peripheral blood cell DNA of 13 (36%) of the 36 women with PBC and in 11 (31%) of the 36 healthy controls. The two groups of PBC patients with and without male DNA sequences were similar in terms of their clinical, biochemical, and serological features. Y-chromosome sequences were found in three of the four PBC women with associated systemic sclerosis. All of the 24 Y-positive samples contained SY154 sequences, but only three PBC patients and six controls showed the presence of both SY154 and SRY sequences. This discrepancy may suggest that not only fetal cells but also fragments of fetal DNA are present in maternal circulation. Overall, our data do not support the hypothesis that fetal microchimerism plays a significant role in the onset or progression of PBC.

Reaktive Sauerstoffspezies (ROS) werden physiologischerweise in Mitochondrien im Rahmen des aeroben Stoffwechsels bei der Reduktion von Sauerstoff zu Wasser gebildet. Im Ejakulat werden ROS vorwiegend aus Leukozyten freigesetzt, aber konnen auch von Spermatozoen selbst gebildet werden. ROS wirken dabei offenbar als physiologische Transmitter zur Vermittlung physiologischer Funktionen wie Kapazitation und Akrosomreaktion. Unphysiologisch hohe ROS-Mengen, wie beim „oxidativen burst“ von Leukozyten gebildet, konnen allerdings diese Funktionen ebenso beeintrachtigen. Weiterhin konnen hohe ROS-Konzentrationen die Lipide der Spermatozoen-Plasmamembranen oxidieren, die wegen der zahlreichen vielfach ungesattigten Fettsauren gegenuber einer Oxidation anfallig sind, und damit die Spermatozoenfunktionen schadigen. Eine erhohte ROS-Bildung im Ejakulat wurde mit einer erhohten DNA-Oxidation in den Spermatozoen, Motilitatsverlust, verminderter Fahigkeit zur Spermatozoen-Oozytenfusion und Infertilitat assoziiert. Die Hauptquelle der ROS im Ejakulat sind die Leukozyten. Als Schutz vor schadigenden Effekten dienen zahlreiche antioxidative Schutzsysteme, die sowohl in der Zelle als auch im Seminalplasma vorhanden sind. Ohne Seminalplasma sind Spermatozoen den durch Leukozyten gebildeten ROS deutlich ungeschutzter ausgesetzt. Diese Situation liegt bei Entzundungen von Hoden oder Nebenhoden sowie bei der Spermatozoenpraparation im Rahmen der assistierten Reproduktion vor. Zur Verhinderung von Schaden sollten daher schonende Verfahren gewahlt werden, bei denen die Leukozyten noch bei Anwesenheit des Seminalplasmas entfernt werden, wie Swim-up vom Samen, Gradienten-Zentrifugation oder Glaswollfiltration. Einzelne Studien sprechen fur gunstige Effekte einer systemischen antioxidativen Behandlung bei mannlicher Infertilitat. Ungeklart bleibt weiterhin die optimale Diagnostik zur Erkennung von Patienten mit einer durch ROS-bedingten Infertilitat sowie die optimale Behandlungsmoglichkeit bezuglich Praparat und Dosis.

To evaluate the tissue-specific distribution of lipophilic antioxidants including various vitamin E forms (tocotrienols and tocopherols) and oxidised and reduced coenzyme Q (ubiquinone and ubiquinol), a sensitive procedure was developed using gradient HPLC with both electrochemical- and UV-detection. A unique distribution of these antioxidants in hairless mouse tissues was found, suggesting that their distribution may be dependent upon selective mechanisms for maintaining antioxidant defences. Ubiquinol-9 was highest in kidney (81 ± 29 nmol/g) and in liver (42 ± 16 nmol/g), while the highest ubiquinone-9 concentrations were found in kidney (301 ± 123 nmol/g) and heart (244 ± 22 nmol/g). Liver contained nearly identical amounts of each ubiquinol-9 (41 ± 16 nmol/g) and ubiquinone-9 (46 ± 18 nmol/g). These mice were fed a commercial chow diet containing α-tocopherol (30 ± 6 mg/kg diet), γ-tocopherol (10 ± 1), a-tocotrienol (3.1 ± 0.7) and γ-tocotrienol (7.4 ± 1.7). Of the vitamin E forms, brain contained only α-tocopherol (5.4 ± 0.1 nmol/g; 99.8%) and no detectable tocotrienols. In other tissues, the α-tocopherol content was higher (20 nmol/g), while each of the other forms represented about 1 % of the total ( γ-tocopherol 0.2 to 0.4 nmol/g, a-tocotrienol 0.1, γ-tocotrienol 0.2). Remarkably, skin contained nearly 15% tocotrienols and 1% γ-tocopherol. The unique distribution of tocotrienols in skin suggested that they might have superior protection against environment stressors. Therefore, the penetration of topically applied vitamin E (tocotrienol enriched fraction of palm oil, TRF) and vitamin E homologue concentrations before and after exposure of skin to UV-light was assessed. 20 μL of 5% TRF in polyethylene glycol-400 (PEG) was applied to 2 skin sites and 20 μL PEG to 2 other sites. After 2 h, the skin was washed and half of the sites exposed to UV-irradiation using a solar simulator (2.8 mW/cm2 for 29 min). The vitamin E content of hairless mouse skin was: α-tocopherol 9.0 ± 1.0 nmol/g skin, γ-tocopherol 0.44 ± 0.03, a-tocotrienol 0.48 ± 0.07, γ-tocotrienol 0.92 ± 0.03. Topical TRF enriched skin vitamin E: α-tocopherol 201 ± 70 nmol/g skin, γ-tocopherol 37 ± 15, a-tocotrienol 53 ± 25, and γ-tocotrienol 50 ± 24. After UV-irradiation, concentrations of all vitamin E homologues from both treatment areas decreased significantly (p0.01), but the TRF-treated skin contained vitamin E at concentrations 7- to 30-fold higher than control values. These findings provide provocative clues on the uptake and regulation of tissue lipophilic antioxidants. The unique distribution of these antioxidant substances suggests their distribution may be dependent upon tissue-specific selective mechanisms.

Reports from North America and Northern Europe have suggested that antimitochondrial antibody (AMA) negative primary biliary cirrhosis (PBC) is a distinct chronic cholestatic liver disease with high prevalence of serum non-organ-specific autoantibodies other than AMA. To evaluate if such a peculiar serum immunoreactivity is associated with clinically relevant characteristics, we reviewed our experience with 297 Italian patients who have had a clinical and histological diagnosis of PBC and were regularly followed-up at our Center from June 1974 to June 1994. AMA-negative and AMA-positive patients were compared in terms of biochemical and clinical features, and clinical outcome of the disease. At presentation, 30 of 297 patients (10%) tested negative for AMA by indirect immunofluorescence. Six of them tested positive for antimitochondrial M2 antibodies (AMA-M2) by immunoblotting analysis, therefore, diagnosis of AMA-negative PBC was made in 24 patients (8%). At the initial visit, AMA-negative and AMA-positive patients were similar in terms of biochemical and clinical features. Antinuclear and anti-smooth-muscle antibodies (ANA and ASMA) were more frequently positive in the AMA-negative patients (71% vs. 31%, and 37% vs. 9%; both P = .0002). Incidence of complications of cirrhosis and development of liver failure resulting in death or referral for liver transplantation did not differ significantly between the two populations. In conclusion, data from this historical cohort study suggest that the distinct serological features of AMA-negative PBC are not associated with substantial differences in the clinical spectrum or course of the disease.

Anthralin is one of the most effective and safest therapeutic agents for the treatment of psoriasis, a skin disease characterized by epidermal hyperproliferation and hyperkeratosis. The drug induces and inflammatory response in the skin involving the expression of cytokine and cell adhesion molecule genes that is thought to be essential for its therapeutic efficacy. Reactive oxygen intermediates (ROIs) generated in vivo during the auto-oxidation of anthralin were discussed as mediators of the inflammatory response, but it is not yet understood how this is translated into novel inflammatory gene expression. In this study, we show that at little as 10 microM anthralin can activate a prototypic form of transcription factor NF-(kappa)B, a central transcriptional regulator of inflammatory and immune responses. Two different lines of evidence show that ROIs, in particular H2O2, are second messengers for the anthralin-induced NF-(kappa)B activation. Firstly, the activation could be inhibited by the structurally unrelated antioxidants N-acetyl-L-cysteine and pyrrolidinedithiocarbamate. Secondly, keratinocytes stably overexpressing catalase showed a significant reduction of NF-(kappa)B activation, while stable overexpression of Cu/Zn-superoxide dismutase augmented the anthralin effect. Our data suggest that ROI-induced NF-(kappa)B plays a role in the anti-psoriatic activity of the drug anthralin.

Liver disease is increasingly recognized as a major cause of morbidity in cystic fibrosis (CF). Preliminary data suggest that ursodeoxycholic acid (UDCA) may be beneficial for treatment of this manifestation. We performed a double-blind, multicenter trial in these patients to establish efficacy and safety of UDCA in terms of the improvement of clinical and nutritional indicators besides standard liver function tests. We also intended to establish whether taurine supplementation has a beneficial effect in patients receiving UDCA. From June to December 1990, we enrolled in 12 centers 55 CF patients with liver disease (39 male subjects; median age, 13.8 years). They were randomly assigned to receive for 1 year one of the following treatments: UDCA (15 mg/kg body weight daily) plus taurine (30 mg/kg body weight daily), UDCA plus placebo, placebo plus taurine, or double placebo. Clinical and laboratory evaluations were performed every 3 months. After 1 year, deterioration of overall clinical conditions, as indicated by the Shwachman-Kulczycki score (SKS), occurred in patients who received placebo but not in those who received UDCA (P = .025). Patients treated with UDCA also showed an improvement in gamma-glutamyl transpeptidase (GGT) (P = .004) and 5'-nucleotidase (P = .006) levels. Treatment with taurine was followed by a significant increase in serum prealbumin levels (P = .053), a trend toward a reduction in fat malabsorption, and no effect on the biochemical profile. No severe side effects occurred with any treatment. Thus, we concluded that UDCA administration improves clinical and biochemical parameters in CF patients with liver disease. Taurine supplementation may be indicated in patients with severe pancreatic insufficiency and poor nutritional status.

BACKGROUND Cure rate for subjects with refractory or relapsing metastatic neuroblastoma is

INTRODUCTION: This study aimed to characterize the in vitro effect of EV-077, acompound that antagonises the binding of prostanoids and isoprostanes to thethromboxane receptor (TP) and inhibits the thromboxane synthase (TS), on plateletaggregation of patients with type-2 diabetes and coronary artery disease (CAD) onchronic aspirin treatment. The effect of EV-077 on 8-iso-PGE(2)-mediated TPreceptor contraction of human arteries was also investigated.MATERIALS AND METHODS: Fifty-two type-2 diabetics with CAD on chronic aspirin(100mg) treatment were studied. Arachidonic acid-induced platelet aggregation wasmeasured by impedance aggregometry in platelet-rich plasma (PRP) and whole blood anticoagulated with hirudin, and by light transmission aggregometry incitrate-anticoagulated PRP following 10-min in vitro exposure to EV-077(100nmol/l) or control. The effect of EV-077 was measured on isometriccontraction of 24 human umbilical arteries induced by isoprostane 8-iso-PGE(2).RESULTS: Arachidonic acid (1mmol/l) induced substantial aggregation inhirudin-anticoagulated whole blood (63±4AU), which was significantly reduced byin vitro exposure to EV-077 (38±3AU, P

The administration of ursodeoxycholic acid, a hydrophilic bile acid not hepatotoxic to humans, has been suggested for treatment of primary biliary cirrhosis to improve cholestasis and reduce hepatocellular damage. Efficacy of treatment has been studied mainly in patients with asymptomatic or early-stage disease. In January 1988, to establish the efficacy and safety of ursodeoxycholic acid in a population with more severe disease, we started a multicenter, double-blind, placebo-controlled trial in patients with symptomatic disease, that is, with pruritus or serum bilirubin exceeding 2 mg/dl. Forty-four patients were assigned to ursodeoxycholic acid, 500 mg daily (corresponding to about 8.7 mg/kg body weight in these patients), and 44 to a placebo. As planned at the beginning of the study, a preliminary analysis was performed when all patients had been followed for at least 6 months (33 patients up to 12 months). Pruritus, self-evaluated by the patients, and cholestyramine consumption, as recorded in a diary, decreased significantly ( p p p p p p

To study the effects of different bile acids on biliary lipids in obese patients with radiolucent gallstones, 12 subjects were given chenodeoxycholic acid (CDCA) at a dose of 15 mg/kg/day, ursodeoxycholic acid (UDCA) at a dose of 15 mg/kg/day, and an equimolar combination of the two (7.5 + 7.5 mg/kg/day) in accordance with a double-blind crossover design. Mean molar percentage of cholesterol and cholesterol saturation index corrected for urso-rich bile (CSI) decreased significantly with all three treatments, but the combination was more effective in decreasing the CSI than either of the two bile acids given alone (p less than 0.05). Bile became desaturated in 10 of 12 patients receiving the combination, in 4 of 12 receiving CDCA, and 3 of 12 receiving UDCA alone. Combination treatment was well tolerated since mild diarrhea and slight increase in transaminases were observed only in a few patients. We conclude that the combined administration of CDCA and UDCA in equimolar doses is the treatment of choice for dissolution of gallstones in obese patients.

Background This study investigates the efficacy and the feasibility of a chemotherapy regimen with topotecan plus vincristine and doxorubicin (TVD) given on an individually tailored basis to patients with refractory/recurrent rhabdomyosarcoma (RMS). Patients and methods Nine patients received TVD therapy at relapse, and six were assessable for response. Results All the six patients experienced objective response after two cycles of chemotherapy: one minor response, four partial response, and one complete response. Conclusions The value of our study is severely limited by the small number of cases, the single-institutional setting and the individually tailored treatment, but we nonetheless confirmed the feasibility and tolerability of topotecan-based chemotherapy in RMS.

Summary Background Dermatofibrosarcoma protuberans (DFSP) is an uncommon tumor of the skin with high rates of local recurrence. It is debated whether Mohs micrographic surgery (MMS) involves lower recurrence rates than wide local excision (WLE). Recent preliminary reports indicate more consistently favorable cure rates with MMS. We report comparative observational data on 41 patients who underwent MMS and 38 who underwent WLE. Their data were then pooled with those available in the medical literature to obtain more precise estimates of recurrence rates with MMS and WLE. Methods The records of 79 patients with DFSP who underwent WLE (n = 38) or MMS (n = 41) in 1990–2005 were reviewed retrospectively. The primary endpoint was tumor recurrence rate. The PubMed database was searched for DFSP case series treated with WLE or MMS, and the recurrence proportions reported for the two separate procedures were pooled. Results Five of the 38 WLE patients (follow-up = 4.8 years) had recurrences (13.2%, 95% CI 4.4–28.1%) as opposed to none (95% CI 0–8.6%) of the 41 MMS patients (follow-up = 5.4 years). Pooling of these data with those from the literature yielded 6/463 recurrences for MMS (1.3%, 95% CI 0.5–2.8%) and 288/1394 recurrences for WLE (20.7%, 95% CI 18.6–22.9%). The relative risk of recurrence for WLE vs. MMS patients was 15.9 (95% CI 7.2–35.5). Conclusions Significantly lower recurrence rates were recorded in our patients subjected to MMS compared with those treated with WLE. The pooled data also indicated a clear advantage of MMS. There is inconclusive evidence for any advantage of MMS in non-primary cases, while MMS was most effective in treating head and neck tumors. These data may be useful to guide clinicians in the choice of the more appropriate surgical treatment for DFSP patients.

PUVA-bath therapy has developed into first line topical PUVA therapy in the treatment of psoriasis. Because of logistical and economic problems, bath PUVA may be difficult to administer. Recently, cream-PUVA therapy has been described as an alternative mode of topical therapy. We treated two patients with moderate plaque-type psoriasis with this new topical approach. 0,0006% 8-methoxypsoralen cream was applied for 1 hour, directly followed by increasing doses of UVA. The number of treatments needed for clearance were 34 and 40. The cumulative UVA dosages were 71.6 and 84 J/cm(2) respectively. Our data document that cream-PUVA therapy is an effective and safe variation of topical PUVA therapy, which may develop into first line photochemotherapy for patients with moderate plaque-type psoriasis.

Metabolic bone disease associated with primary biliary cirrhosis (PBC) is inadequately characterized. Renal tubular acidosis (RTA) may lead to bone loss through chronic mobilization of skeletal calcium salts to buffer increased acid load.To evaluate the prevalence of RTA in PBC and establish the relationships among bone mineral density (BMD), renal function and nutritional status.We enrolled 69 female patients with compensated PBC and 35 control patients with chronic hepatitis C. RTA was searched in all patients, and 24-h dietary recalls were collected at enrolment. BMD was measured by dual-energy X-ray absorptiometry at the femur neck, lumbar spine and radius ultradistalis sites.No patients received a diagnosis of RTA. BMD values (Z-scores) showed only little deviation from normal population with no difference between PBC and controls. Osteopoenic PBC patients (T-score1) showed significantly lower daily phosphorus intake [median: 672 (288-1374) vs. 921 (253-1923) mg/day; P = 0.037], with a trend towards lower caloric intake than their nonosteopoenic counterparts.Renal tubular acidosis is uncommon in compensated PBC. Cholestasis is not associated with an increased risk of bone demineralization. Inadequate dietary intake may be a preventable factor contributing to bone loss in PBC.

Serum antibodies to carbonic anhydrase (CA) II have been reported in patients with autoimmune pancreatitis (AIP) and Sjogren's syndrome (SjS). However, their significance in the pathogenesis of these diseases is controversial. The aim of this study was to identify serum antibodies to CA isozymes, which are expressed in ductal cells of the pancreas.Recombinant proteins of human CAs IV, IX, and XII were obtained using a bacterial expression system, and five CA IV peptides with theoretically high antigenicity were synthesised. Western blotting and enzyme linked immunosorbent assay (ELISA) were used to detect serum antibodies to the CA isozymes.The first screening analysis by western blot showed serum antibodies to CA IV among three CA isozymes in patients with idiopathic chronic pancreatitis, including AIP patients. Further analysis by ELISA showed a significantly increased prevalence of serum antibodies to the truncated CA IV protein and the CA IV synthetic peptide (LGS LTT PTC DEK VVW TVF REP I) in patients with definite AIP (4/15 and 6/20, respectively; p0.01), probable AIP (6/14 and 3/14; p0.02), and SjS (9/20 and 8/40; p0.001) compared with normal controls (0/26). There was no significant difference in the antibody prevalence rates between normal controls and patients with alcoholic chronic pancreatitis (2/15 in each) or pancreatic cancer (2/14 and 1/14, respectively). The presence of serum antibodies to the CA IV peptide showed significant correlations with serum gamma-globulin and IgG levels in AIP patients.These findings suggest that CA IV may be a target antigen that is commonly expressed in epithelial cells of specific tissues involved in AIP and its related diseases.

The aim of the present study was to evaluate the effectiveness of two consecutive nonrandomised treatment programs applied between 1989 and 1999 at the Istituto Nazionale Tumori of Milan in an unselected cohort of 59 children over the age of one with stage 4 neuroblastoma. Both treatment programs consisted of two phases, the induction of the remission phase and the consolidation phase. The induction of the remission phase consisted of intensive chemotherapy, and remained the same throughout the study period. The consolidation phase consisted of sequential hemi-body irradiation (HBI) (10 Gy per session, 6 weeks apart) in the first period (1988-June 1994) and sequential high-dose cyclophosphamide, etoposide, mitoxantrone+L-PAM and autologous haemopoietic stem cell transplantation in the second (July 1994-1999). Intention-to-treat analysis revealed a significantly better outcome for patients treated with the second program, the 5-year event-free survival probability being 0.12 for program 1 and 0.31 for program 2 (P=0.03). This finding led us to conclude that sequential HBI is useless as consolidation treatment. The high-dose chemotherapy adopted in the second program enabled a proportion of patients to obtain long-term survival but, since the clinical results remain unsatisfactory, new treatment strategies are warranted.

The skin is directly and frequently exposed to a pro-oxidative environment, including ozone and UV-radiation. While ozone in the stratosphere protects against mutagenic UVC-radiation, it is also a major air pollutant in urban areas. With its strong oxidizing potential, ozone is perhaps one of the most reactive chemicals the skin ever encounters. Although a large body of evidence exists for ozone- induced oxidative stress in the respiratory tract, the current knowledge on its in vivo effect on cutaneous tissues is based on studies of the last 10 years. Acute ozone exposure damages the stratum corneum, depletes skin vitamin C and E and induces lipid and protein oxidation in upper epidermal layers. Secondary products penetrate into deeper skin layers and are capable of activating signal transduction pathways and inducing cell damage. It has been shown in a murine model, that environmentally relevant ozone concentrations can induce a stress response in the skin.

Lymphocyte migration is an essential requirement for efficient surveillance of tissues for infectious pathogens and for recruitment of effector cells at sites of injury or infection. Although leukocyte recirculation functions in immune surveillance, excessive trafficking to extravascular locations can lead to serious tissue injury and destruction [1, 2, 3]. A role of inappropriate T-cell skin homing, via induction of T-cell cutaneous lymphocyte-associated antigen (CLA),has recently been suggested in the initiation and perpetuation of the inflammatory process of skin diseases [4, 5, 6]. CLA is a ligand for E-selectin expressed on human lymphocytes which can be induced by bacterial superantigens such as TSST-i [6, 7, 8]. It is characterized by its reactivity with the antibody HECA-452 and expressed on the maj ority of T-cells in skin. Given the very initial role of carbohydrate-selectin interactions in the emigration process and the potential of excessive leukocyte extravasation for tissue injury, inhibition of carbohydrate-selectin interaction seems to be a promising tool for therapeutic intervention in inflammatory processes [2,9].

Laparoscopy with laparoscopic ultrasound (L-LUS) has proved to be superior to conventional CT imaging in the staging of hepatocellular carcinoma (HCC). The aim of our prospective study was to evaluate the efficacy of L-LUS as compared with currently available imaging techniques (spiral CT or Lipiodol CT) in patients with HCC and liver cirrhosis. From January 1998 to May 2000, 70 consecutive patients (50 men and 20 women; mean age 67 +/- 7 years) were enrolled. Liver cirrhosis was related to chronic hepatitis C virus infection in 55, hepatitis B virus infection in seven, and alcohol abuse in eight patients. Preoperative diagnostic workup included the following: 70 ultrasound examinations of the liver, 23 CT scans after Lipiodol arteriography, and 53 spiral CT scans. A single lesion was found in 39 patients, two lesions in 20 patients, and three lesions in 11 patients. L-LUS was performed in all patients under general anesthesia using a two- to three-trocar technique. The examination was completed in 68 patients (97%); in two cases extensive adhesions prevented the L-LUS examination. L-LUS yielded additional information in 39 patients (57%). New histologically proved HCC lesions were detected in 14 patients (in the same liver segment in 4 cases and in different liver segments in 10 cases), and an adrenal metastasis was seen in one patient. In 23 patients, benign nodules were identified as regenerative macronodules, low-grade dysplastic nodules, or small hemangiomas. In 10 patients, correct localization of the primary lesion was detected by L-LUS in comparison with the preoperative liver location. In our experience, L-LUS is a safe and reliable procedure. It provides superior information (intraoperative histologic confirmation) for the diagnosis and pretreatment staging of HCC in patients with cirrhosis when compared with current radiologic imaging techniques.

Few studies have assessed the significance of prognostic factors in cirrhotic patients undergoing resection for hepatocellular carcinoma.Overall survival and disease-free survival were evaluated in 53 cirrhotic patients undergoing hepatic resection for supervening hepatocellular carcinoma. The value of the UICC TNM classification, and the Izumi modified staging system on prognosis were analyzed.In multivariate analysis lack of micro/macrovascular invasion were predictive for long-term outcome. The difference between stages 1 and 2 or stage 3 and 4A using the UICC TNM classification was not significant with respect to survival or disease-free survival. UICC TNM classification was modified as follows; stage 1, solitary tumor without vascular invasion; stage 2, solitary or multiple tumor(s) involving adjacent vessel branch; stage 3, tumor(s) involving major vessel branch or with regional lymph node metastases; stage 4, tumor(s) with distant metastases. TNM (modified in accordance with Izumi) showed a significant difference between each stage with respect to survival and disease-free survival.A uniform tumor classification of hepatocellular carcinoma is advocated. Our results show that the UICC TNM classification for hepatocellular carcinoma is inadequate and may even on occasion lead to unnecessary resection. The modified staging system of Izumi is superior in determining outcome for cirrhotic patients with supervening hepatocellular carcinoma undergoing resection.

In this report, the feasibility and efficacy of laparoscopic radiofrequency interstitial thermal ablation of hepatocellular carcinoma has been evaluated in 20 patients.The laparoscopic approach with the use of intraoperative ultrasonography allowed us to obtain additional information regarding liver nodules and a complete treatment of the liver lesions.The complication rate was low and there was no operative mortality. A complete necrosis has been obtained in 90% of the cases at 1 month dynamic computed tomography following the treatment.Laparoscopic radiofrequency thermal ablation of hepatocellular carcinoma proved to be a safe and effective technique; its use may be proposed in selected patients. Larger series are needed to accurately assess its role among the other ablative therapies of hepatocellular carcinoma.

Many studies have demonstrated beneficial health effects of topical antioxidant application; however, the underlying mechanisms are not well understood. To better understand the protective mechanism of oxogenous anti-oxidants, it is important to clarify the physiological distribution, activity and regulation of antioxidants. Also, the generation of ROS by the resident and transient microbial flora and their interaction with cutaneous antioxidants appears to be of relevance for the redox properties of skin. Our studies have demonstrated that alpha-tocopherol is, relative to the respective levels in the epidermis, the major antioxidant in the human SC, that alpha-tocopherol depletion is a very early and sensitive biomarker of environmentally induced oxidation and that a physiological mechanism exists to transport alpha-tocopherol to the skin surface via sebaceous gland secretion. Furthermore, there is conclusive evidence that the introduction of carbonyl groups into human SC keratins is inducible by oxidants and that the levels of protein oxidation increase towards outer SC layers. The demonstration of specific redox gradients within the human SC may contribute to a better understanding of the complex biochemical processes of keratinization and desquamation. Taken together, the presented data suggest that, under conditions of environmentally challenged skin or during prooxidative dermatological treatment, topical and/or systemic application of antioxidants could support physiological mechanisms to maintain or restore a healthy skin barrier. Growing experimental evidence should lead to the development of more powerful pharmaceutical and cosmetic strategies involving antioxidant formulations to prevent UV-induced carcinogenesis and photoaging as well as to modulate desquamatory skin disorders.

Combined medical treatment may provide further benefit to primary biliary cirrhosis (PBC) patients administered ursodeoxycholic acid (UDCA).To evaluate the long-term effects of colchicine and UDCA in symptomatic PBC patients.We extended up to 10 years the double-blind treatment of 44 symptomatic PBC patients originally included in a 3-year multicentre study comparing UDCA and colchicine (U + C) versus UDCA and placebo (U + P). Outcome measures were death or liver transplantation; incidence of clinically relevant events; clinical and quantitative variables retaining prognostic information.Mean follow-up was 7 +/- 3 years. One patient was lost, three withdrew because of jaundice (U + P); two patients stopped colchicine but remained on UDCA. Eleven patients (two for liver-unrelated reasons, U + P) and six patients (U + C) died, three and two patients, respectively, were transplanted (incidence rate difference, five cases per 100 patient-years; 95% CI, -1 to 11). Hepatocellular carcinoma developed in one (U + P) and four (U + C) patients (difference, -2; CI, -5 to 1), portal hypertension complications in nine patients from each group (difference, 1; CI, -5 to 6). Trends of serum bilirubin, Mayo score, antipyrine clearance were similar among treatment groups.In cirrhotic PBC patients, colchicine does not offer additional benefits to UDCA. In this population, UDCA does not obviate disease progression.

To establish the efficacy of combination therapy with ursodeoxycholic acid (UDCA) and colchicine in patients with symptomatic primary biliary cirrhosis (PBC), defined by the presence of liver cirrhosis, pruritus or bilirubin exceeding 2 mg/mL.A total of 90 patients were randomly assigned to ursodeoxycholic acid 500 mg/daily plus placebo (UDCA group, n=44), or ursodeoxycholic acid at the same dosage plus colchicine, 1 mg/daily (UDCA/C group, n=46). The two groups were comparable for age, sex, stage of disease, severity of pruritus, bilirubin, and Mayo score. All patients underwent clinical, ultrasonographic, and biochemical examinations at entry and then every 6 months up to 3 years of follow-up. Patients with cirrhosis underwent endoscopy every 12 months. In a sub-group of patients without cirrhosis, who consented, liver biopsy was repeated at the end of the study.The number of treatment failures (i.e. dead, orthotopic liver transplantation (OLT), complications of cirrhosis, doubling of bilirubin, untreatable pruritus) was 11 (25%) in the UDCA group and four (9%) in the UDCA/C group (P0.05). No significant differences were observed in terms of improvement of liver enzymes related to cholestasis and cytolysis and of amelioration of pruritus. The Mayo score values increased less above the baseline values at 24 and 36 month-intervals in the UDCA/C group than in the UDCA group. Histological evaluation at baseline and at the end of the study was available for 15 patients with pre-cirrhotic stage. A significant reduction in histological grading score was observed in patients from the UDCA/C group, whereas no changes in these histological scores were observed in the UDCA group.The addition of colchicine to ursodeoxycholic acid in patients with symptomatic primary biliary cirrhosis results in a small but significant reduction of disease progress.

To assess risk factors for gallstone recurrence following non-surgical treatment.A prospective follow-up of a multicentre cohort of post-dissolution gallstone patients.Six gastroenterology units in the UK and Italy.One hundred and sixty-three patients with confirmed gallstone dissolution following non-surgical therapy (bile acids or lithotripsy plus bile acids), followed up by ultrasound scan and clinical assessment at 6-monthly intervals for up to 6 years (median, 25 months; range, 6-70 months).Subject-related variables (sex, age, height, weight, body mass index), gallstone-related variables (number, diameter, presence of symptoms, months to complete stone clearance), treatment modalities (bile acid therapy, extracorporeal shock wave lithotripsy) and follow-up related variables (weight change, use of non-steroidal anti-inflammatory agents, statins, pregnancies and/or use of oestrogens) were assessed by univariate and multivariate analysis as putative risk factors for gallstone recurrence.Forty-five gallstone recurrences were observed during the follow-up period. Multiple primary gallstones and length of time to achieve gallstone dissolution were the only variables associated with a significant increase in the recurrence rate. Appearance of biliary sludge during follow-up was also significantly related to development of gallstone recurrence. Use of statins or non-steroidal anti-inflammatory agents did not confer protection against recurrence.Patients with primary single stones are the best candidates for non-surgical treatment of gallstones, because of a low risk of gallstone recurrence. The positive association of recurrence with biliary sludge formation and time to dissolution of primary stones may provide indirect confirmation for the role of impaired gallbladder motility in the pathogenesis of this condition.

Ultraviolet (UV-) radiation therapy as a mono- or combination therapy (UV-A, UV-A1, UV-B, SUP, UV-B311) or as photochemotherapy with photosensitization (systemic PUVA-, bath PUVA-, topical PUVA-therapy) are successfully used for the treatment of several dermatological disorders. Long-term side effects of natural UV (sun light) include photoaging and induction of skin tumors. At present, the relevance of in-vitro findings of potential tumor induction in animals through therapeutic levels of UV radiation is a matter of debate. To assess these risks, information on treated location, kind of UV radiation and cumulative UV doses are required. Practically this information is barely accessible. This makes decisions on possible therapies difficult. To solve this problem we propose to use an "UV pass". At the end of each UV radiation cycle, the body location, the type of radiation and the cumulative dose are documented and this pass is given to the patient. This will improve the information transfer if the doctor is changed, as well as facilitating decisions about certain therapies and calculation of long-term risks of UV radiation. Finally it will improve the quality of UV photo- and photochemotherapy.

Acute and severe impairment of liver function with jaundice and ascites occurred in two out of seven patients with chronic hepatitis D during interferon alpha administration (10 MU three times a week). Both of them were young women with histological diagnoses of moderate to severe chronic hepatitis and cirrhosis with no signs of portal hypertension. Only a slow and partial recovery was observed after interferon withdrawal. Autoantibodies against basal cell layer tested positive in these two patients. In the remaining five patients with hepatitis D who did not experience liver impairment during interferon administration, basal cell layer antibodies were found only in one case. We conclude that severe decompensation of liver cirrhosis related to hepatitis D may occur during interferon administration. Positivity of basal cell layer antibodies may be associated with the risk of developing such an adverse event but our data are not sufficient to prove this association.

SUMMARYAntibody to carbonic anhydrase II, an enzyme abundantly present in biliary epithelium, has been proposed as a diagnostic marker for antimitochondrial antibody-negative PBC. In this study we determine its prevalence and clinical significance in a large series of patients with antimitochondrial antibody-positive and -negative PBC. Reactivity to carbonic anhydrase II was sought by Western immunoblotting in sera from 215 consecutive patients with PBC (26 antimitochondrial antibody-negative), 13 with autoimmune hepatitis, 25 with primary Sjögren's syndrome (pSS), 12 with systemic sclerosis, 19 with systemic lupus erythematosus and 73 healthy subjects. The prevalence of antibody to carbonic anhydrase II (titre 1:100) in PBC was 8%. No specific reactivity to carbonic anhydrase II was found in antimitochondrial antibody-negative PBC (7% versus 8% in antimitochondrial antibody-positive PBC). Ascites (P = 0.006) and Sjögren's syndrome (SS) (P = 0.022) in PBC were significantly associated with presence of the antibody. In patients with SS associated with PBC, the prevalence (19%) was similar to that observed in pSS (16%). At a serum dilution of 1:40, the prevalence of positive sera in PBC rose to 27% but disease specificity was reduced. Our findings in a large population of PBC patients rule out a relation between presence of antibody to carbonic anhydrase II and lack of antimitochondrial antibody. The higher prevalence of ascites found in positive patients warrants further evaluation.

To evaluate skin penetration of various vitamin E homologs, a 5% solution of either alpha-tocopherol, alpha-tocotrienol, or gamma-tocotrienol in polyethylene glycol was topically applied to SKH-1 hairless mice. After 0.5, 1, 2, or 4 h (n = four per time point and four per vitamin E homolog), the skin was washed, the animals killed, the skin rapidly removed, frozen on dry ice, and a biopsy taken and sectioned: stratum corneum (two uppermost, 5-micron sections--SC1 and SC2), epidermis (next two 10-micron sections--E1 and E2), papillary dermis (next 100 microns, PD), dermis (next 400 microns, D), and subcutaneous fat (next 100 microns, SF). SC1 contained the highest vitamin E concentrations per mu thickness. To compare the distribution of the various vitamin E forms into the skin layers, the percentage of each form was expressed per its respective total. Most surprising was that the largest fraction of skin vitamin E following topical application was found in the deeper subcutaneous layers--the lowest layers, PD (40 +/- 15%) and D (36 +/- 15%), contained the major portion of the applied vitamin E forms. Although PD only represents about 16% of the total skin thickness, it contains sebaceous glands--lipid secretory organs, and, thus, may account for the vitamin E affinity for this layer. Hence, applied vitamin E penetrates rapidly through the skin, but the highest concentrations are found in the uppermost 5 microns.

As the outermost barrier of the body, the skin is directly and frequently exposed to a prooxidative environment, including solar UVA and UVB radiation, and air pollution. Ozone, a major air pollutant, possesses a strong oxidizing potential and therefore is possibly one of the most reactive chemicals the skin ever meets. Although a large body of evidence exists for ozone induced oxidative stress in the respiratory tract, the current knowledge of its in vivo effect on cutaneous tissues is very limited. Acute ozone exposure depletes skin vitamins C and E and induces lipid peroxidation in upper epidermal layers. The stratum corneum, as the penetration barrier of the body, appears to be particularly susceptible to ozone induced oxidative stress. Such processes at superficial skin layers lead to barrier perturbations and could trigger inflammatory responses in adjacent skin layers. Furthermore, there is some in vitro evidence for an involvement of ozone in cutaneous inflammation and carcinogenesis. Taken together, these findings may be particularly relevant for photochemical smog: concomitant exposure to UV-irradiation and ozone could reveal synergistic oxidative stress effects in skin. The mechanisms of ozone induced effects in cutaneous tissues are still poorly understood. Further studies in this young field of research are needed to evaluate its relevance for skin disorders in humans.

An association between celiac disease and primary biliary cirrhosis has been reported in a few cases, mainly as individual case reports.To screen adult patients with celiac disease for primary biliary cirrhosis and patients with primary biliary cirrhosis for intestinal celiac involvement.The celiac group consisted of 336 adults (218 women and 118 men; mean age, 36 yr; range 18-74 yr) with celiac disease diagnosed by serological and histological tests, 38 with newly diagnosed celiac disease and 298 with previously diagnosed celiac disease who were consuming a gluten-free diet. The mean follow-up period was 6 yr (range, 1-16 yr). Liver function parameters and autoantibody levels were determined, and, when indicated, histological tests were performed. The biliary cirrhosis group consisted of 65 subjects (58 women and seven men) (mean age, 59 yr; range, 35-67 yr) with primary biliary cirrhosis diagnosed 1-17 years previously (mean, 7 yr) on the basis of the usual biochemical, serological, and histological criteria. Antigliadin and antiendomysium antibody levels were determined, and two biopsy specimens from the distal duodenum obtained during endoscopy were evaluated.In patients with celiac disease, impairment of liver function was frequently found at diagnosis (16 of 38, or 44%), but primary biliary cirrhosis was diagnosed in only one case. In patients with primary biliary cirrhosis, no cases of celiac disease, as currently defined, were found.Our findings indicate that celiac disease and primary biliary cirrhosis are rarely associated and support the hypothesis that the intestinal lesions per se are not responsible for the liver disease.

To assess the relationship between changes in liver histology and virological parameters of HCV infection in patients with a sustained biochemical response to alpha-interferon treatment for chronic hepatitis, with the aim of identifying the most reliable indicator of treatment efficacy.Quantitative serial testing of HCV-RNA in plasma samples collected on a monthly basis over the last period of post-treatment follow-up were tested in sixteen subjects withor = 12 months aminotransferase normalisation following discontinuation of interferon therapy. The quantitative HCV-RNA determination in plasma was performed by a competitive reverse-transcription polymerase-chain reaction. Pretreatment and 12-month post-treatment liver histologies were blindly evaluated using a semi-quantitative scoring system. At these times a qualitative HCV-RNA analysis was also carried out.The post-treatment histological grading score was significantly reduced. Nevertheless, all patients except one tested positive for HCV-RNA in plasma (mean 1.7 x 10(6) molecules/ml): nine were positive in all the serial determinations, while the remaining 6 were intermittently positive. No relationship between genotype, viral load, pattern of viremia (intermittent or continuous) and change in histological score was found.Changes in liver histology are the most reliable indicator of the efficacy of interferon treatment in hepatitis C related liver disease. HCV-RNA clearance based on serial sampling does not appear to be a reliable indicator, even in the presence of a sustained biochemical response and histological improvement.

Background/Aims: Following non-surgical treatment, cholesterol gallstones recur in a high proportion of patients, and recurrence cannot be predicted nor effectively prevented. Our aim was to test prospectively the viability and the efficacy of repeated bile acid therapy, in which recurrent stones are diagnosed at an early stage by regular ultrasound monitoring and promptly retreated, as a strategy for the management of these patients in clinical practice. Methods: One hundred and seventy-two consecutive patients were recruited upon achieving complete gallstone dissolution using non-surgical therapy (bile acids or lithotripsy plus bile acids), and followed up at 6-monthly intervals by ultrasound scan. Gallstone recurrence was promptly treated by a combination of ursodeoxycholic acid plus chenodeoxycholic acid (5 mg/kg per day each) for a period of 2 years, or less if complete redissolution was achieved. Median follow-up period was 34 months (range 6–70). Results: Forty-five patients had gallstone recurrence; of these, 39 underwent one or more repeated courses of bile acid therapy (follow-up data available in 27). Gallstone recurrence rate was 15% at 1 year and 47% at 5 years. Average annual redissolution rate of recurrent gallstones (intention to treat) was 41%. The proportion of gallstone-free patients in the whole population was 88%, 84%, 77%, 78%, 75% at 1–5 years, respectively, and rose to > 90% at 3 years onwards in patients with single primary stones. Conclusions: We conclude that repeated bile acid therapy maintains the majority of patients gallstone free, and is there fore an effective long-term management strategy, especially in patients with primary single gallstones.