Clean Colon Software Program (CCSP), Proposal of a standardized Method to quantify Colon Cleansing During Colonoscopy: Preliminary Results

Background and study aims: Neoplastic lesions can be missed during colonoscopy, especially when cleansing is inadequate. Bowel preparation scales have significant limitations and no objective and standardized method currently exists to establish colon cleanliness during colonoscopy. The aims of our study are to create a software algorithm that is able to analyze bowel cleansing during colonoscopies and to compare it to a validate bowel preparation scale. Patients and methods: A software application (the Clean Colon Software Program, CCSP) was developed. Fifty colonoscopies were carried out and video-recorded. Each video was divided into 3 segments: cecum-hepatic flexure (1st Segment), hepatic flexure-descending colon (2nd Segment) and rectosigmoid segment (3rd Segment). Each segment was recorded twice, both before and after careful cleansing of the intestinal wall. A score from 0 (dirty) to 3 (clean) was then assigned by CCSP. All the videos were also viewed by four endoscopists and colon cleansing was established using the Boston Bowel Preparation Scale. Interclass correlation coefficient was then calculated between the endoscopists and the software. Results: The cleansing score of the prelavage colonoscopies was 1.56\ub10.52 and the postlavage one was 2,08\ub10,59 (P<0.001) showing an approximate 33.3% improvement in cleansing after lavage. Right colon segment prelavage (0.99\ub10.69) was dirtier than left colon segment prelavage (2.07\ub10.71). The overall interobserver agreement between the average cleansing score for the 4 endoscopists and the software pre-cleansing was 0.87 (95% CI, 0.84\u20130.90) and post-cleansing was 0.86 (95% CI, 0.83\u20130.89). Conclusions: The software is able to discriminate clean from non-clean colon tracts with high significance and is comparable to endoscopist evaluation

Predizione del sovraccarico marziale mediante una nuova tecnica di spettrometria di massa per il dosaggio dell'epcidina plasmatica

Background. Hepcidin (Hep) has emerged as the primary regulator of iron homeostasis. Previous studies on assessing urinary levels of Hep are of limited availability. We have developed a new method for quantifying Hep in plasma by SELDI-TOF mass spectrometry, using the 25-AA peptide as reference standard. Aims: 1) to assess the performance of this new method in different conditions of iron metabolism disorders; 2) to assess the diagnostic validity of non invasive serum markers in identifying iron overload. Methods: the following groups of subjects were enrolled into the study: 1. type I hemochromatosis (HE)(n=10), NAFLD (n=17), chronic hepatitis C (n=10), healthy controls - previously enrolled in a general population epidemiological study - with normal ultrasound, normal LFTs, alcohol assumption <20 g ethanol/day, and negative for C282 mutations (n=155). The following parameters were assayed in each case: plasma Hep, C282Y and H63D mutations of the HFE gene by (Taqman chemistry); serum iron, ferritin (SF), transferrin saturation (TfSat), transaminases, GGT, glucose, insulin, total cholesterol, HDL-cholesterol, LDL-cholesterol, triglycerides. Results: Plasma Hep levels were significantly higher in HCV+ (26.3 + 7.2 nmol/L) pts compared to controls (12.3+ 1.0)(one-way ANOVA: F=3.2, p<0.05), and were positively correlated with SF (r=0.451, p<0.001). H63D heterozygous subjects revealed a pattern of iron overload (significantly higher serum iron, SF, TfSat, and lower Hep/SF ratio) compared to H63D wild type subjects. By analysing data with the Biomarker Pattern 5.0.2. Software, in order to identify the most significant discriminant markers between HE and controls, we obtained a four-terminal node algorithm which included as main splitters Hep/SF ratio, glucose and iron. These variables allowed a correct diagnosis of HE with a 100% sensitivity, 98.6% specificity and AUROC=0.993. Conclusions: The new plasma Hep mass spectrometry method yields accurate measurements which reflect pathologic and genetic influences; simple non invasive markers (Hep/SF ratio, glucose and iron) can predict the presence of HE.Introduzione. L'Epcidina (Hep) si ritiene essere il principale regolatore dell'omeostasi del ferro. Precedenti studi sul dosaggio urinario dell'Hep sono limitati. Abbiamo sviluppato un nuovo metodo per quantificare Hep nel plasma attraverso la spettrometria di massa SELDI-TOF, utilizzando come standard di riferimento il peptide di 25-AA. Obiettivi. 1) valutare la performance di questa nuova metodica in diverse condizioni di alterazione del metabolismo del ferro; 2) valutare la validità diagnostica di una marker sierico non invasivo per identificare il sovraccarico di ferro. Metodi. I seguenti gruppi di soggetti sono stati arruolati nello studio: 1. Emocromatosi di tipo I (HE) (n=10), NAFLD (n=17), epatite C cronica (n=10), controlli sani - precedentemente reclutati in uno studio epidemiologico sulla popolazione generale - con ecografia epatica non patologica, funzionalità epatica normale, introito alcolico <20 g etanolo/die, e negativi per la mutazione C282 (n=155). I seguenti parametri sono stati saggiati in ciascun caso: Hep plasmatica, mutazioni C282Y e H63D del gene HFE attraverso sonde Taqman; ferro sierico, ferritina (SF), satuazione della transferrina (TfSat), transaminasi, GGT, glucosio, insulina, colesterolo totale, colesterolo HDL, colesterolo LDL, trigliceridi. Risultati. I livelli plasmatici di Hep sono risultati significativamente più elevate nei soggetti HCV+ (26.3 + 7.2 nmol/L) rispetto ai controlli sani (12.3+ 1.0) (one-way ANOVA: F=3.2, p<0.05), e sono risultati positivamente correlati con SF (r=0.451, p<0.001). I soggetti eterozigoti per H63D hanno dimostrato un pattern di sovraccarico di ferro (livelli significativamente più elevati di ferro sierico, SF, TfSat, e un più basso rapporto Hep/SF) rispetto ai soggetti H63D-wild type. Dall'analisi dei dati con il Software Biomarker Pattern 5.0.2., per identificare il marker discriminante più significativo tra HE e i controlli, abbiamo ottenuto un algoritmo a 4 nodi che include come splitter principali il rapporto Hep/SF, il glucosio ed il ferro. Queste variabili sono sufficienti per diagnosticare correttamente HE con il 100% sensibilità, il 98.6% di specificità e AUROC=0.993. Conclusioni. La nuova applicazione della spettrometria di massa per dosare i livelli di Hep plasmatici fornisce misure accurate che rilevano basi patologiche e genetiche; semplici marker non invasivi (Hep/SF, glucosio e ferro) possono predire la presenza di HE

Pegylated interferon alpha-2b plus ribavirin for naive patients with HCV-related cirrhosis

BACKGROUND: Data on the efficacy of antiviral therapy in patients with HCV-related compensated cirrhosis are generally drawn from analyzing subgroups in larger trials. AIMS: (1) To analyze the safety and efficacy of combination therapy in naive patients with HCV-related cirrhosis; (2) to evaluate the factors influencing the sustained virologic response (SVR) in cirrhotic patients by comparison with a group of noncirrhotic patients; (3) to analyze the outcome of cirrhotic patients either acquiring SVR and nonresponders to the antiviral therapy during the posttreatment follow-up. METHODS: We consecutively enrolled 365 patients with biopsy-proven HCV-related chronic hepatitis meeting the inclusion criteria for pegylated interferon a-2b plus Ribavirin: 87 patients had compensated liver cirrhosis and 278 had histologic stages between 1 and 4 according to Ishak's classification. RESULTS: The 2 groups were comparable for genotype, viral load, and alanine transferase at presentation. Cirrhotic patients were significantly older and had significantly higher body mass index, serum ferritin, and gamma-glutamyl transpeptidase. The rate of side effects was similar in the 2 groups, whereas the rate of SVR was significantly lower in cirrhotic (45.9%) than in noncirrhotic patients (65.8%). Logistic regression analysis showed that genotype 1 to 4 and high viral load were independent variables correlating with nonresponse in the sample as a whole. During follow-up, hepatocellular carcinoma developed in 5/38 (13.2%) cirrhotic patients not responding or relapsing after treatment. No cases of hepatocellular carcinoma were seen among cirrhotic or noncirrhotic patients with a SVR. CONCLUSIONS: Cirrhotic patients with compensated disease have a reasonably good chance of virologic response and should be offered treatment, carefully monitoring any side-effects

Intron 2 [IVS2, T-C +4] HFE gene mutation associated with S65C causes alternative RNA splicing and is responsible for iron overload

A patient with congenital liver fibrosis revealed a high transferrin saturation index and iron overload on liver biopsy. He did not carry the most frequent HFE mutations: C282Y or H63D. Heterozygosity was detected for S65C. Unknown HFE mutations were also sought using a combined denaturing high performance liquid chromatography (DHPLC)/direct sequence approach and another point mutation, a transition T-C (nt 4910), at the fourth base of the donor splice site of intron 2 [HFE, intervening sequence (IVS) 2, T-C +4] was found. Family screening revealed that a daughter carried both S65C and [IVS2, T-C +4]

Hepatobiliary phospholipid transporter ABCB4, MDR3 gene variants in a large cohort of Italian women whit intrahepatic cholestasis of pregnancy

12noIntrahepatic cholestasis of pregnancy is a multifactorial disorder of pregnancy associated with a genetic background.To evaluate the genetic contribution of ABCB4, MDR3 gene in the development of intrahepatic cholestasis of pregnancy in a large cohort of Italian subjects.This study represents an extension of a previous multicentre-prospective study including three Italian referral centres. In all, we enrolled 96 women at the 3rd trimester of pregnancy. Genomic DNA was extracted from peripheral venous blood leucocytes by standard procedures. Polymerase chain reaction was used to amplify exon 14, 15 and 16 of MDR3 gene.We found 3 non-synonymous heterozygous mutations in exon 14 (E528D, R549H, G536A), 1 in exon 15 (R590Q) and 2 in exon 16 (R652G, T6671). MDR3 gene variants in exons 14, 15 and 16 occurred in 7/96 of pregnant mothers with intrahepatic cholestasis of pregnancy (7.2%), and in none of 96 pregnant controls matched for age and parity. All seven patients had normal gamma-glutamyl transpeptidase, normal bilirubin, but high levels of both alanine transferase and serum bile acids. One had cholesterol biliary lithiasis. The outcome of pregnancy was normal in four cases (with vaginal delivery), while there was one fetal distress.MDR3 mutations are responsible for the development of intrahepatic cholestasis of pregnancy in only a small percentage of Italian women. Further genetic studies are warranted, however, to clarify the role of different mutations in intrahepatic cholestasis of pregnancy.reservedmixedAZZAROLI F.; BRAGHIN C.; CARDERI L.; ESPOSITO W.; FLOREANI A.; MARCHESONI Diego; MAZZELLA G.; MONTAGNANI M.; PATERNOSTER D.; ROSA RIZZOTTO E.; SOARDO GIORGIO; VARIOLA A.Azzaroli, F.; Braghin, C.; Carderi, L.; Esposito, W.; Floreani, A.; Marchesoni, Diego; Mazzella, G.; Montagnani, M.; Paternoster, D.; ROSA RIZZOTTO, E.; Soardo, Giorgio; Variola, A

Plasma Adiponectin Levels in Primary Biliary Cirrhosis: A Novel Perspective for Link Between Hypercholesterolemia and Protection Against Atherosclerosis

INTRODUCTION: Hypercholesterolemia is a common finding in primary biliary cirrhosis (PBC), but the risk of cardiovascular events in PBC patients is not increased in respect to the general population. High serum adiponectin levels appear to play a protective role in the development of either metabolic syndrome or cardiovascular disease. AIM: To investigate factors potentially preventing atherosclerosis in PBC patients. METHODS: Circulating levels of adiponectin, resistin, leptin, and tumor necrosis factor-alpha (TNF-alpha) were measured in 137 consecutive PBC patients (125 women, 12 men; mean age 61.6 +/- 12.3 yr), 137 sex- and age-matched healthy controls, and 30 female patients with nonalcoholic steatohepatitis (NASH) and associated metabolic syndrome. RESULTS: The body mass index (BMI) was comparable in the three groups, whereas total cholesterol was significantly higher in both PBC and NASH cases than in controls (221.6 +/- 50.5 mg/dL in PBC vs 221.7 +/- 39.7 mg/dL in NASH vs 209.8 +/- 39.2 mg/dL in controls, P < 0.05). Serum concentrations of adiponectin, resistin, and leptin were significantly higher in PBC patients than in either NASH cases or controls (P < 0.05). Among the PBC patients, only adiponectin correlated positively with histological progression of the disease (P= 0.001) and negatively with BMI (P= 0.01). Logistic regression analysis revealed that adiponectin correlated independently with age, BMI, Mayo score, and gamma-glutamyl transpeptidase. CONCLUSIONS: The high adiponectin concentrations observed in PBC patients should be regarded as a possible protective factor against atherogenesis. The search for further protective factors should be encouraged

Tumor Budding as a Risk Factor for Nodal Metastasis in Pt1 Colorectal Cancers: A Meta-Analysis

Worldwide, colorectal cancer (CRC) screening programs have significantly increased the detection of sub-mucosal (pT1) adenocarcinoma. Completion surgery may be indicated after endoscopic excision of these potentially metastasizing early cancers. However, the post-surgical prevalence of nodal implants does not exceed 15%, leading to questions concerning the clinical appropriateness of any post-endoscopy surgery. Eastern scientific societies (Japanese Society for Cancer of the Colon-Rectum, in particular) include tumor budding (TB), defined as the presence of isolated single cancer cells or clusters of fewer than five cancer cells at the tumor invasive front, among the variables that must be included in histological reports. In Western countries, however, no authoritative endorsements recommend the inclusion of TB in the histology report due to the heterogeneity of definitions and measurement methods as well as its apparent poor reproducibility. To assess the prognostic value of TB in pT1-CRCs, this meta-analysis evaluated 41 studies involving a total of 10,137 patients. We observed a strong association between the presence of TB and risk of nodal metastasis in pT1-CRC. In comparing TB-positive (684/2,401; 28.5%) versus TB-negative (557/7,736; 7.2%) patients, the prevalence of nodal disease resulted in an OR value of 6.44 (95%CI: 5.26-7.87; p<0.0001; I2=30%). This increased risk of regional nodal metastasis was further confirmed after accounting for potential confounders. These results support the priority of histologically reporting TB in any endoscopically removed pT1-CRC to direct more appropriate patient management