The efficacy of pantoprazole

magnesium was compared with

The efficacy of pantoprazole

magnesium was compared with that of esomeprazole over 4 and 8 weeks’ treatment in patients with erosive gastroesophageal reflux disease (GERD). Methods: In this multicentre (14 Brazilian sites), phase III, double-blind study, patients with erosive GERD (Los Angeles grades A–D) were randomised to pantoprazole magnesium (n = 290) or esomeprazole (n = 288), both administered at 40 mg once daily for 8 weeks. Severity of esophagitis (at endoscopy) and GERD-related symptoms (using ReQuest™-GI) were assessed at baseline and 4 and 8 weeks, and complete remission (a ReQuest™-GI score below 1.73 Wnt inhibitor plus endoscopically confirmed healing) was compared between treatments (significance p < 0.05). Results: Complete remission with pantoprazole magnesium was non-inferior to that with esomeprazole at 4 and 8 weeks (table). Mucosal healing was similar for the two treatments. However, symptom relief with pantoprazole magnesium was significantly greater at 8 weeks (p = 0.0370) than that with esomeprazole. Both PPIs had similarly low rates of adverse events. Conclusion: Pantoprazole magnesium 40 mg was as effective as esomeprazole 40 mg for complete remission and mucosal healing, but provided greater symptom relief at 8 weeks, suggesting an

extended period of treatment effect. Key Word(s): 1. GERD; 2. Symptom relief; 3. Pantoprazole; 4. Esomeprazole; Complete remission, endotcopically confirmed healing and symptom relief rates after 4 and 8 weeks (intent-to-trcat population) Assessment 4 weeks, n (%) 8 weeks, n(%) Pantoprazole magnesium Esomeprazole buy NVP-LDE225 Pantoprazole magnesium Sitaxentan Esomeprazole *P = 0.0370 versus esomeprazole. Presenting Author: RAPAT PITTAYANON Additional Authors: RATHA-KORN VILAICHONE, TANISA PATCHARATRAKUL, CHINNAVAT SUTTHIVANA, WANIT PIYANIRUN, MONTIRA MANEERATANAPORN, SOMCHAI LEELAKUSOLVONG,

UDOM KACHINTORN, SUTEP GONLACHANVIT, VAROCHA MAHACHAI Corresponding Author: RAPAT PITTAYANON Affiliations: Chulalongkorn University; Thammasart University; Bhumipol Adulyadej Hospital; Pramongkulklao Hospital; Siriraj Hospital, Mahidol University Objective: Introduction: The awareness of gastroesophageal reflux disease (GERD) has led to an increased prevalence of GERD across the Asian region. It has a significant impact on quality of life and health care expenditure. Proton pump inhibitor (PPI) is commonly used to relief the symptoms. The aim of this study was to understand the patient perception on GERD, its impact on quality of life and the pattern of PPI use in GERD patients in Thailand. Methods: Methods: The physician-diagnosed GERD patients recruited from hospitals throughout Thailand participated in the 20 minute face-to-face interviews after signing informed consent. Results: Results: A total of 400 patients from 39 hospitals were enrolled.

e , class III phosphatidylinositol-3-kinase) The process of auto

e., class III phosphatidylinositol-3-kinase). The process of autophagosome formation involves two major steps: nucleation and elongation of the isolation membrane. The Atg1/unc-51-like

kinase (ULK) complex, the autophagy-specific PI3-kinase complex, and PI3P-effectors and their related proteins are important for the nucleation p38 MAPK activity step, whereas the Atg12- and LC3/Atg8-conjugation systems are important for the elongation step. Studies have demonstrated that autophagy plays a wide variety of physiological and pathophysiological roles. Recent evidence has shown that autophagy is associated with cancer pathogenesis and that pharmacologic manipulation of autophagic pathways may represent a new therapeutic strategy for human cancers. However, to date the role of autophagy in cancer is not clearly defined. Although autophagy is a cancer cell survival mechanism against environmental and cellular stresses, it can be associated with cancer cell death under certain situations. Further,

autophagy and apoptosis might be linked to each other and occur simultaneously or sequentially in a cell type-, death stimulus-, and context-dependent manner.[9] Although Hh signaling is known to inhibit cell apoptosis, it remains unknown whether Hh signaling is able to regulate autophagy. The current study describes a novel role of the Hh signaling pathway for regulation of autophagy in human HCC cells. We show that inhibition of the Hh pathway markedly enhanced autophagy http://www.selleckchem.com/products/CP-673451.html through up-regulation of Bnip3 (a member of BH3-only subset of the Bcl-2 family) that displaces Bcl-2 from its binding partner, Beclin-1, and that this process preludes apoptosis. Our findings suggest that the status of autophagy (autophagic flux) is a key factor that may influence cell response to Hh-targeted therapy. Human HCC cells (Huh7, Hep3B, and HepG2) were treated with the Hh signaling ligand, agonists, or inhibitors as indicated and the cells were analyzed for autophagy by immunoblotting

for microtubule-associated Selleckchem Rucaparib protein light chain 3 (LC3) and p62, GFP-LC3 puncta, monodansylcadaverine (MDC) staining, and transmission electron microscopy (TEM). Western blotting analysis was performed to determine the alteration of key signaling molecules including Shh, Patched, Smo, and Gli1, Bnip3, Bcl-2 family proteins, and MEK/ERK1/2. The interaction between Bcl-2 and Beclin-1 was analyzed by immunoprecipitation and immunoblotting assays. For quantitative reverse-transcription polymerase chain reaction (qRT-PCR), total RNA was extracted with the TRIzol plus RNA purification kit and reverse-transcribed to complementary DNA (cDNA) using Superscript II reverse transcriptase; the cDNA samples were used for real-time PCR analysis in triplicate using the QuantiFast SYBR Green PCR kit (Qiagen) on a Bio-Rad C1000 Thermal Cycler.


“This chapter contains sections titled: Introduction Princ


“This chapter contains sections titled: Introduction Principles of biologic standardization Standardization of factor VIII assays Standardization of factor IX assays Standardization of inhibitor assays Standardization of von Willebrand factor assays Standardization of bypassing agents

Standardization of assays of other coagulation factors Standardization of global assays References “
” Twenty years ago I conceived an idea for a journal about haemophilia. I approached Peter Saugman of Blackwell Publishing – a company that was particularly strong in Haematology: their first scientific publication, the British Journal of Haematology, was published in 1955. Fortunately, they were prepared to take the risk, and wanted the journal to be international and have a strong North American presence. Doreen Brettler, director of the New England Hemophilia Centre, agreed to become the first North American editor. We met to formulate our ideas and develop MLN0128 molecular weight an editorial board at the World AIDS Meeting in Berlin in June 1993. It was important from the outset to have the support of key haemophilia leaders – Shelby Dietrich, then the head of the World Federation of Hemophilia (WFH) publications committee, and Pier Mannucci provided helpful support and advice. Peter Jones, the director of the Newcastle Haemophilia Centre in the

UK, encouraged us to present the idea at a WFH meeting of the ‘Decade Plan’ held in Estoril, Portugal in October 1993. With BGB324 cost some trepidation, at the end of a long meeting,

I presented a mock-up of the first cover with the title Haemophilia – there was no discussion, even about the anglicized Greek spelling! The launch issue appeared in October 1994 with a publication date January 1995. Our mission statement that ‘Haemophilia is an international journal dedicated to the exchange of information regarding the comprehensive care of haemophilia’ Cyclic nucleotide phosphodiesterase continues today. The journal soon became the official journal of WFH and proudly published, for the first time, the abstracts of the WFH meeting held in Dublin 1996. More recently Haemophilia has become affiliated to the European Association for Haemophilia and Allied Disorders (EAHAD) and the Hemostasis and Thrombosis Research Society of North America (HTRS). We publish in translation a Japanese edition and a Chinese edition. Haemophilia is now published by Wiley-Blackwell and is one of their 1500 scientific publications. We continue to publish predominantly in print, but an increasing number of our readers prefer on-line; the authors are also fast moving in this direction as this issue of Haemophilia attests. Haemophilia remains grateful to all the Authors who have submitted, and continue to submit, their research, writing and thinking – together we have created a substantial record of haemophilia, and the many challenges concerning the management of this intriguing condition.

These results may have been, in part, reflective of the imbalance

These results may have been, in part, reflective of the imbalance in BMI between the groups in that study, as well as differences in environmental and dietary factors between the two study cohorts. In addition, the lack of classification of patients in each group based

on liver histology may have also affected the results of the study by Zhu et al.29 Lastly, differences in age may have also played a role, as discussed below. There are various theories to support an inverse correlation between Bacteroidetes and steatohepatitis. First, Bacteroidetes carry 45% of the lean metabolic potential in a study comparing the microbiome of lean and obese adults.21 A lower percentage of Bacteroidetes could have affected energy balance by facilitating metabolic dominance of other bacteria

that are more efficient in extracting energy from the diet. Jumpertz et al.11 showed that a 20% increase in fecal Bacteroidetes is associated with a 150 kcal decrease in energy harvest from the diet. A second theory is that an initial hit causes the cell death of Bacteroides leading to LPS release from their cell wall and subsequent endotoxemia.19 The latter leads to the development of NASH.19, 27 It is not clear what would cause the death of these microorganisms. Changes in diet could play a role, as shown by studies in obese subjects, whose baseline lower fecal Bacteroidetes increase when placed on a hypocaloric diet or after bariatric mafosfamide surgery.38, 39 There is literature supporting an increase in intestinal permeability of subjects with IR, such as in obesity40 and diabetes.41 Recently, Zhu et al.29 reported higher serum ethanol levels in children

with NASH, which was thought to be bacterially derived and, hence, potentially also contributing to increased intestinal permeability. In addition, there is scientific evidence linking Trichostatin A cost endotoxemia with states of glucose intolerance, such as NAFLD.41, 42, 43 Animals and humans exposed to low levels of endotoxin develop IR.19, 22 Exploratory analysis from our study also suggested a potential link between the intestinal microbiota and IR by showing a trend toward a negative association between Bacteroidetes and IR when controlling for BMI. This requires further studies. We did not find lower bifidobacteria counts or higher Firmicutes-to-Bacteroidetes ratio in NASH compared to SS and HC. This is in contrast to some of the previously published literature in the field of obesity9, 14, 44 and the recent study by Zhu et al. on children with NASH.29 The inability to show differences in these bacteria may have been due to the sample size; however, the size of our cohort was similar to that of other cross-sectional studies on IM in obesity9, 11, 12, 37 and NAFLD.29 Our results on Firmicutes-to-Bacteroidetes ratio are in line with other smaller projects, which also failed to replicate the findings of Ley et al.

86; 95% CI 076–098; P value for the trend = 0039) (Table 2) T

86; 95% CI 0.76–0.98; P value for the trend = 0.039) (Table 2). This

study clearly indicated that higher serum levels of direct bilirubin are significantly associated with a lower risk of developing NAFLD. Nevertheless, application of these results to the general population of either sex remains controversial because NASH was associated with a significantly decreased prevalence of unconjugated hyperbilirubinemia,[41] and this Korean study was limited to men. UA is the final oxidation product of purine catabolism, and hyperuricemia is considered a metabolic disease; many studies have also reported a relationship between hyperuricemia and NAFLD. Li et al. examined the relationship between ICG-001 mw UA levels and NAFLD in a cross-sectional study among 8925 company Wnt inhibitor employees (6008 men) and showed that hyperuricemia,

as well as male gender, age, BMI, WC, GGT level, TG level, HDL-c level, low-density lipoprotein-cholesterol level, and fasting plasma glucose (FPG), was an independent risk factor for NAFLD (OR 1.291; 95% CI 1.067–1.567; P < 0.001) in multiple regression analysis (Table 1).[16] Sirota et al. conducted a cross-sectional analysis of 10 732 non-diabetic adults who participated in the National Health and Nutrition Examination Survey 1988–1994 in the United States.[17] They defined sex-specific UA quartiles (≤ 5.2, 5.3–6.0, 6.1–6.9, and > 6.9 mg/dL for men and ≤ 3.7, 3.8–4.5, 4.6–5.3, and > 5.3 mg/dL for women) this website and revealed that the OR for the highest quartile was 1.43 (95% CI 1.16–1.76, P < 0.001) compared

with the lowest quartile after adjusting for demographic data, hypertension, WC, TG level, HDL-c level, HOMA-IR, estimated glomerular filtration rate (eGFR), and AST level (Table 1). In addition, Hwang et al. studied 9019 Korean individuals who visited a health checkup center and had UA levels within the normal range. These patients were categorized into four groups according to UA quartiles for both sexes, and the relationship between the UA level and the presence of NAFLD was examined.[18] After adjusting for age, smoking status, regular exercise, BMI, BP, FPG, TC level, TG level, HDL-c level, AST level, ALT level, and GGT level, the adjusted ORs (95% CIs) for the presence of NAFLD in the subjects with the highest UA level was 1.46 (1.17–1.82) for men and 2.13 (1.42–3.18) for women as compared with the subjects with the lowest UA level (Table 1).

1D) These findings suggest

that HMGB1 may, in fact, be a

1D). These findings suggest

that HMGB1 may, in fact, be actively released into the circulation of patients with HCC. Hypoxia is a hallmark of solid tumors, including HCC.8 Accordingly, we found hypoxia-inducible factor-1 alpha levels to be substantially increased in liver homogenates of HCC specimens, compared to nontumor tissue (Supporting Fig. 1). Immunofluorescence Sirolimus ic50 showed that HMGB1 mainly localized in the nucleus of hepatocytes in the nontumorous liver. However, in addition to nuclear HMGB1, cytoplasmic HMGB1 was also present at high levels in HCC cells (Fig. 1E). To determine whether hypoxia plays a role in inducing the expression or translocation of HMGB1 in HCC cells, Hepa1-6 and Huh7 cells were cultured under normoxic (21% O2) or hypoxic (1% O2) conditions. Under normoxia in both cell lines, HMGB1 remained located predominantly in the nucleus. After exposure

to hypoxia, the expression of HMGB1 increased in the cytoplasm Bortezomib chemical structure (Fig. 1F). These findings indicate that hypoxia leads to the nuclear to cytoplasmic translocation of HMGB1 in HCC cells. To determine whether hypoxia induces the expression or translocation of HMGB1 in HCC, western blotting analysis was performed. The expression of HMGB1 in whole cell lysates was not significantly changed during hypoxia at either the protein or mRNA level (Fig. 2A; Supporting Fig. 2). In contrast, hypoxia led to a time-dependent increase of HMGB1 translocation to the cytoplasm and HMGB1 release into the media in both cell lines (Fig. 2B,C). In addition, cell viability was not substantially different, when

comparing HCC cells exposed to hypoxia and normoxia, as assessed in a crystal violet viability assay (Fig. 2D). These findings demonstrate that hypoxia leads to the translocation and release of HMGB1 in HCC cells. Caspases play an important role in programmed cell death and inflammation.16, 19 Though apoptosis inducers caspase-3 and -9 were activated in our hypoxic HCC cells (data not shown), we focused on caspase-1, which can induce inflammation and affect tumor progression. To characterize caspase-1 activation during hypoxia, Hepa1-6 and Huh7 cells were learn more cultured under hypoxic conditions for various times. The 45-kDa procaspase-1 was cleaved into the active 20-kDa caspase-1 during hypoxia and was increased in a time-dependent manner during hypoxia (Fig. 3A,B). Using a colorimetric assay to assess caspase-1 activity during hypoxia, we found that caspase-1 activity was also significantly increased in both Hepa1-6 and Huh7 cells subjected to hypoxia, compared to normoxic controls (Fig. 3C). To confirm the localization of cleaved caspase-1, we performed immunofluorescent staining for cleaved caspase-1. After hypoxia, cleaved caspase-1 significantly increased in the cytoplasm diffusely in both cell lines (Supporting Fig. 3). These findings demonstrate that hypoxia induces caspase-1 activation in HCC cells.

674, P = 0879) and age (F = 0902, P = 0445) distribution

674, P = 0.879) and age (F = 0.902, P = 0.445) distribution

were not statistically different.21 cases of EST large incision group, 2 cases of gallbladder developing (9.5%), no development in 19 cases (90.5%); 20 this website cases of EST medium and small incision group, 17 cases of gallbladder developing (85%), no development in 3 cases (15%); 20 cases of EPBD group, 16 cases of gallbladder developing (94.1%), no development in 1 case (5.9%); 20 cases of control group, 19 cases of gallbladder developing (95%), no development in 1 case (5%). There was significant difference between the large incision group and the others groups such as control group, small incision group, EPBD group (P < 0.001); but compared with the control group, both EST small incision and EPBD group had no the significant difference (P = 0.609 和 P = 0.598); there was no significant difference between EST small incision group and EPBD group (P = 1.000). Campared the gallbladder development time (GT) and emptying fraction GBEF (%) at the 30th minute among the medium and small incision EST group, EPBD group and control group.

The median of gallbladder development time GT (minimum and maximum) was 32 (30, 60) minutes in EST medium and small incision group. The median of gallbladder development time GT (minimum and maximum) was 32.5 (30, 50) minutes in EPBD group. The median of gallbladder development time GT (minimum and maximum) was 30 (30, 35) minutes in control group. The gallbladder development time and emptying fraction GBEF (%) was no significant difference among three Selleckchem Galunisertib groups. Conclusion: The research of this subject proves the integrity relationship between Oddi sphincter and gallbladder function from a new angle. It provides related theoretical foundation for the future clinical selection of ERCP operation and the prevention of long-term complications. The purpose is to protect the Oddi sphincter and the normal gallbladder function, reduce short and long-term complications while taking out the common bile duct calculi.

Key Word(s): 1. ERCP; 2. gall bladder; 3. EST; 4. EPBD; Presenting this website Author: DENNISNYUK FUNG LIM Additional Authors: ISHFAQ AHMAD Corresponding Author: DENNISNYUK FUNG LIM Affiliations: KETTERING GENERAL HOSPITAL NHS TRUST; ALEXANDRA HOSPITAL, WORCESTER ACUTE HOSPITAL NHS TRUST Objective: Endoscopic retrograde cholangio-pancreatography (ERCP) is a technically difficult procedure that is associated with a substantial risk of complications and may result in death. Awareness of these problems has led to recommendations designed to restrict the number of procedures by careful selection of patients and the use of alternative methods of investigation. However, there is evidence that patients continue to be subjected inappropriately to ERCP. A recent extensive audit by the British Society of Gastroenterology 3 indicated that approximately 48,000 ERCPs are performed each year in the United Kingdom.

0 ± 015; HFHFr, 16 ± 029; ATRA + HFHFr, 21 ± 017), albeit no

0 ± 0.15; HFHFr, 1.6 ± 0.29; ATRA + HFHFr, 2.1 ± 0.17), albeit not to a statistically significant degree.

These results suggest that ATRA also normalizes insulin sensitivity in the diet-induced NAFLD mice, possibly by reversing leptin resistance. We examined retinoid signaling in the livers of NAFLD mice, in which ligand-dependent RAR-mediated transcriptional regulation plays a central role. Although hepatic RARα expression was not changed, expression of the RARα target gene Rarb was significantly down-regulated in HFHFr mice, whereas ATRA reversed this effect (Supporting Fig. 6A-C). This result was consistent with our previous observation that hepatic retinoid signaling is impaired in NAFLD patients22 Pritelivir cost and

prompted us to perform transcriptome analysis using complementary DNA microarrays. By identifying genes with elevated expression in the ATRA + HFHFr group compared with the HFHFr group, four independent probes corresponding to Lepr were ranked as the highest 10 (Table 1). This finding was consistent with the leptin-dependent effect of ATRA on insulin sensitivity. qPCR confirmed significant up-regulation of LEPRa as well as IGF binding protein 2 (IGFBP2), which is expressed in the liver in response to systemic leptin administration29 (Supporting Fig. 6D,E). This finding suggests that the leptin-signaling pathway was activated in the livers of mice fed the ATRA + HFHFr RG7204 mouse diet. We next examined the expression of leptin-signaling proteins. Consistent with

the qPCR data, Western blotting revealed that the expression of the short LEPR isoform was also significantly higher in the ATRA + HFHFr mice compared with that in the HFHFr selleck inhibitor group (Fig. 4E, Supporting Fig. 5C). Note that LEPRb protein expression was not detected in liver samples. Although the total JAK2 level was lower in HFHFr and ATRA + HFHFr group mice than in controls, its phosphorylation was significantly elevated in the latter group (Fig. 4E, Supporting Fig. 5D,E). Total and phosphorylated STAT3 expression were significantly up-regulated by ATRA, whereas suppressor of cytokine signaling 3 (SOCS3) was not (Fig. 4E, Supporting Fig. 5F-H). STAT3 activation in hepatocytes by ATRA was also demonstrated by immunohistochemistry, showing intense nuclear staining of STAT3 in hepatocytes throughout the liver lobule in the ATRA + HFHFr group, while STAT3 was distributed diffusely in the cytoplasm and nucleus of pericentral hepatocytes in the control and HFHFr groups (Supporting Fig. 7). No changes were observed in the levels of other leptin signaling molecules, adenosine monophosphate-activated protein kinase α, or extracellular signal-regulated kinases 1/29, 30 (data not shown).

3%) had NAFLD, 13 (224%) had chronic cryptogenic liver disease,

3%) had NAFLD, 13 (22.4%) had chronic cryptogenic liver disease, 5 (8.6%) had AIH, 5 had nodular regenerative hyperplasia (NRH) of the liver,

4 (6.8%) had CDG, 2 (3.44%) had congenital hepatic fibrosis, and 2 (3.44%) had Klippel-Trénaunay-Weber syndrome with hepatic vascular malformations. Furthermore, celiac disease, chronic hepatitis C, Alagille syndrome, and sclerosing cholangitis were each present in a single case. The remaining six patients were recruited after familial screening and did not carry any mutation according to the molecular analysis of ATP7B. Roscovitine mouse Liver function tests and other routine laboratory data were obtained with standard methods. The ceruloplasmin concentration in serum was measured by radial immunodiffusion (NOR-Partigen Coeruloplasmin, Behring, Marburg, Germany; normal range = 20-60 mg/dL).12 Urine samples (basal urinary learn more copper and urinary copper after PCT) were collected in an acid-washed, plastic, metal-free container. PCT urinary copper was evaluated after patients ingested 500 mg of D-penicillamine at time zero and again at

12 hours while 24-hour urinary copper collection progressed.13 Copper levels in urine were determined by flame atomic absorption spectrophotometry as previously described.14 Liver biopsy was performed by the Menghini technique with a disposable biopsy set (Hepafix, Braun, Melsungen, Germany). Copper levels in dried liver tissue were determined by flame atomic absorption spectroscopy according to Kingston and Jassie15 (normal selleckchem range = 6-50 μg/g of dry weight). All slides were examined by the same pathologist, and lesions were evaluated according to the recommendations of Batts and Ludwig.16 For the molecular analysis of the ATP7B gene, DNA extraction and polymerase chain reaction were carried out with the standard methods by Dr.

Georgios Loudianos (Ospedale Regionale per le Microcitemie, Cagliari, Italy). With single-strand conformational polymorphism and sequencing methods, patients were analyzed for the 12 exons (5, 6, 8, 10, and 12-19) on which most mutations reside according to previous studies of the Italian continental population. DNA samples not completely characterized by the first step of analysis or those found to have a new missense mutation were further analyzed for the remaining exons of the ATP7B gene by single-strand conformational polymorphism and sequencing analysis.17 Continuous variables (ceruloplasmin, urinary copper, and liver copper) were presented as numbers of patients, means, medians, and standard deviations, whereas discrete variables (clinical manifestations at presentation and the presence or absence of KF rings) were presented as percentages. Normally distributed continuous variables were presented as means and standard deviations and were compared between groups by analysis of variance with post hoc testing (Scheffe’s test).

3%) had NAFLD, 13 (224%) had chronic cryptogenic liver disease,

3%) had NAFLD, 13 (22.4%) had chronic cryptogenic liver disease, 5 (8.6%) had AIH, 5 had nodular regenerative hyperplasia (NRH) of the liver,

4 (6.8%) had CDG, 2 (3.44%) had congenital hepatic fibrosis, and 2 (3.44%) had Klippel-Trénaunay-Weber syndrome with hepatic vascular malformations. Furthermore, celiac disease, chronic hepatitis C, Alagille syndrome, and sclerosing cholangitis were each present in a single case. The remaining six patients were recruited after familial screening and did not carry any mutation according to the molecular analysis of ATP7B. FK506 supplier Liver function tests and other routine laboratory data were obtained with standard methods. The ceruloplasmin concentration in serum was measured by radial immunodiffusion (NOR-Partigen Coeruloplasmin, Behring, Marburg, Germany; normal range = 20-60 mg/dL).12 Urine samples (basal urinary selleck copper and urinary copper after PCT) were collected in an acid-washed, plastic, metal-free container. PCT urinary copper was evaluated after patients ingested 500 mg of D-penicillamine at time zero and again at

12 hours while 24-hour urinary copper collection progressed.13 Copper levels in urine were determined by flame atomic absorption spectrophotometry as previously described.14 Liver biopsy was performed by the Menghini technique with a disposable biopsy set (Hepafix, Braun, Melsungen, Germany). Copper levels in dried liver tissue were determined by flame atomic absorption spectroscopy according to Kingston and Jassie15 (normal check details range = 6-50 μg/g of dry weight). All slides were examined by the same pathologist, and lesions were evaluated according to the recommendations of Batts and Ludwig.16 For the molecular analysis of the ATP7B gene, DNA extraction and polymerase chain reaction were carried out with the standard methods by Dr.

Georgios Loudianos (Ospedale Regionale per le Microcitemie, Cagliari, Italy). With single-strand conformational polymorphism and sequencing methods, patients were analyzed for the 12 exons (5, 6, 8, 10, and 12-19) on which most mutations reside according to previous studies of the Italian continental population. DNA samples not completely characterized by the first step of analysis or those found to have a new missense mutation were further analyzed for the remaining exons of the ATP7B gene by single-strand conformational polymorphism and sequencing analysis.17 Continuous variables (ceruloplasmin, urinary copper, and liver copper) were presented as numbers of patients, means, medians, and standard deviations, whereas discrete variables (clinical manifestations at presentation and the presence or absence of KF rings) were presented as percentages. Normally distributed continuous variables were presented as means and standard deviations and were compared between groups by analysis of variance with post hoc testing (Scheffe’s test).