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BACKGROUND: Innovative approaches to improve diagnostic yield of endoscopic ultrasound-guided tissue acquisition (EUS-TA) have focused on needle design with development of fine-needle biopsy (FNB) needles with microcore-acquisition technology. Recently, a 20-gauge (20G) antegrade-cutting-side-bevelled biopsy needle (ProCore®) was developed for EUS-TA, but data about its diagnostic performance and histological capability are scant. OBJECTIVES: We assessed the diagnostic performance and histologic retrieval rate of a new 20G antegrade-cutting-side-bevelled biopsy needle compared with a 22G reverse-side-bevelled needle for EUS sampling of solid pancreatic lesions. PATIENTS AND METHODS: A retrospective analysis of 238 consecutively collected patients who underwent EUS-TA using a 20G or a 22G ProCore® needle, without rapid on-site evaluation (ROSE), was conducted at two centres. Sensitivity, specificity, positive predictive value and negative predictive value were calculated. Histologic tissue retrieval was evaluated applying a scoring system for each case. RESULTS: Sensitivity and specificity were estimated as 98.4–100% in the 20G-, and 94.9–100% in the 22G-needle groups, respectively (p > 0.99). The 20G procured more histologic-grade tissues (92.6% vs 49.5%, p

Pediatric malaria, even in countries considered as free as Italy, is an important problem of public health because children have a high variability of the clinical picture. The objective of this brief note is to determine the incidence of pediatric malaria in the Piedmont Region during the period 1989-2015.All cases of pediatric malaria notified were considered thanks to the regional information flow over the period 1989-2015. Cases of congenital malaria, unconfirmed malaria cases, and aged 14 and older were excluded of the study.In Piedmont in the period 1989-2015, pediatric malaria accounts for 8.8% of the total (172/1946 cases). 74% of patients are of foreign nationality, to which must be added the 14% represented by those born in Italy from foreign parents, while it is 100% the fraction of patients who have made a trip to the abroad. The notification of cases is greater in the autumn months. Only 7.6% of the sample carried out a complete chemoprophylaxis. In 79% of cases, the primary care physician advised chemoprophylaxis on trips to endemic areas.At present, lacking an effective vaccine, the prevention and implementation of standard precautions such as chemoprophylaxis, represent the safest strategy to put into practice to eradicate the disease especially for the groups at greater risk as visiting friends and relatives.

Quantitative bone marrow (BM) MR sequences, as DWI and CSI, were used to evaluate BM water–fat composition. The aim of the study was to assess the potential usefulness of fat fraction (FF) and ADC, calculated by CSI or DWI, in diagnosing and classifying myeloma (MM) patients according to their different BM infiltration patterns. The study group included 43 MM patients (19F; 24M; mean age 64 years), 15 asymptomatic, 15 symptomatic with diffuse BM infiltration and 13 symptomatic with focal lesions (FLs). The control group was made up of 15 healthy subjects (7F; 8M; mean age 64 years). MRI examinations consisted of sagittal T1w TSE on the spinal column, axial DWI (b 50–400–800 mm2/s) and coronal T2 Dixon, on the whole body. Mean ADC and FF were calculated placing 1 ROI on 6 vertebras and 2 ROIs on either the pelvis or FL. ANOVA with Bonferroni’s correction showed a significant difference in ADC values among the different groups of MM patients (P

BACKGROUND Small cell lung cancer (SCLC) is a highly aggressive neoplasm that accounts for approximately 10% to 15% of lung cancers. In most cases, the diagnosis relies on cytology and needs to be confirmed by immunohistochemistry. Although several genetic and molecular abnormalities have been recorded, molecular markers able to predict the prognosis are still lacking. MicroRNA (miRNA) signatures have been recently proposed as useful biomarkers in lung cancer because of their high stability during standard sample processing. METHODS Cytological samples for 50 patients with SCLC were collected from primary tumors (n = 25) and metastases (n = 25) by means of fine-needle aspiration (FNA) or bronchial washing (BW); they were fixed in ethanol (FNA) or Duboscq-Brazil fluid (BW). The 3-miRNA panel expression (miR-192, miR-200c, and miR-205) was quantified with a TaqMan polymerase chain reaction miRNA assay and was compared with overall survival (OS) and clinicopathological data. RESULTS All samples had sufficient RNA for the miRNA expression analysis to be performed, regardless of the sample source or the fixative medium. Patients with a low expression level of the 3-miRNA panel were associated with better OS in univariate (P = .032) and multivariate analyses (P = .022). Moreover, in the group of patients older than the mean age of our cohort (65.8 years), a significant OS advantage (P = .013) was seen for patients with a low expression level of the 3-miRNA panel. CONCLUSIONS A specific 3-miRNA signature is potentially useful for predicting survival for patients with SCLC, and it may be feasible with cytological samples taken during standard diagnostic procedures. Cancer Cytopathol 2016. © 2016 American Cancer Society.

AimsFluorescence in situ hybridisation (FISH) increases the sensitivity for detecting pancreatobiliary tract cancer over routine cytology. In this study, diagnostic accuracy and costs of cytology and FISH in detecting cancer in patients with jaundice with biliary strictures were assessed.MethodsBrushing specimens from 109 patients with jaundice were obtained during endoscopic retrograde cholangiopancreatography and examined by cytology and FISH. The specimens were considered FISH-positive for malignancy if at least five polysomic cells or 10 cells with homozygous or heterozygous 9p21/p16 deletion were detected. Definitive diagnosis of the stricture as benign or malignant relied on surgical pathology (45 cases) or clinical-radiological follow-up >18 months (64 cases). We calculated costs of cytology and FISH based on the reimbursement from the Piedmont region, Italy (respectively, €33 and €750).ResultsNinety of 109 patients had evidence of malignancy (44 pancreatic carcinomas, 36 cholangiocarcinomas, 5 gallbladder carcinomas, 5 other cancers), while 19 had benign strictures. Routine cytology showed 42% sensitivity, but 100% specificity for the diagnosis of malignancy, while FISH-polysomy showed 70% sensitivity with 100% specificity and FISH-polysomy plus homozygous or heterozygous 9p21/p16 deletion showed 76% sensitivity with 100% specificity. The cost per additional correct diagnosis of cancer obtained by FISH, in comparison with cytology, was €1775 using a sequential cytological approach (ie, performing FISH only in patients with negative or indeterminate cytology).ConclusionsFISH should be recommended as the second step in detecting cancer in patients with jaundice with pancreatobiliary tract strictures and cytology negative or indeterminate for malignancy.

AIMS High mobility group box 1 (HMGB1) is a chromatin structural protein, expressed ubiquitously in the nuclei of mammalian cells. When transported extracellularly, it acts as a tumour suppressor and oncogenic protein. In malignant pleural mesothelioma (MPM), high serum levels of HMGB1 have been related to a poor prognosis. Conversely, the significance of HMGB1 expression in MPM tissues is still unclear. METHODS AND RESULTS Biopsy samples from 170 patients with MPM were assessed by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) to evaluate HMGB1 protein and gene expression. The expression level of HMGB1 protein was scored using a semiquantitative system that sums the intensity (0-3) and the percentage (from 0 to 4) of positively stained cells in nuclei, cytoplasm and in both. The final score was considered as high (>3) or low (

Background Endoscopic ultrasonography is accurate, safe, and cost-effective in diagnosing common bile duct stones, thus suggesting the possibility to avoid invasive endoscopic retrograde cholangiopancreatography. Aim To prospectively evaluate the diagnostic and therapeutic performance of early endoscopic ultrasonography in suspected choledocholithiasis. Patients and methods All consecutive patients presenting to the Emergency Department with suspicion of choledocholithiasis between January 2010 and January 2012 were evaluated and categorized as low, moderate, or high probability of choledocholithiasis, according to accepted criteria. Endoscopic endosonography was carried out within 48 h from the admission and endoscopic retrograde cholangiopancreatography was performed soon in case of confirmed choledocholithiasis. Results Overall 179 patients were included: 48 (26.8%) were classified as low, 65 (36.3%) as moderate, and 66 (36.9%) as high probability of choledocholithiasis. Of the 86 patients with common bile duct stones at endoscopic endosonography, endoscopic retrograde cholangiopancreatography confirmed the finding in 79 (92%). By multivariate analysis only the common bile duct diameter proved an independent predictor of common bile duct stones. Conclusions Early endoscopic endosonography is accurate in identifying choledocholithiasis allowing immediate endoscopic treatment and significant spare of unnecessary endoscopic retrograde cholangiopancreatography. This approach can be useful as a triage test to select patients not needing endoscopic retrograde cholangiopancreatography, allowing, in selected cases, their early discharge.

Summary Endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) is the procedure of choice for the cytologic diagnosis of pancreatic masses. The specificity of EUS-FNA approaches 100%, but the sensitivity is still low, and the high rate of indeterminate (atypical and suspicious) and false-negative results needs improvement. KRAS gene is frequently mutated in pancreatic ductal adenocarcinoma (PDAC) (up to 90%), and mutation analysis of KRAS has been proposed as diagnostic biomarker of PDAC. In most laboratories, KRAS mutation testing is performed by Sanger sequencing or real time-quantitative polymerase chain reaction (RT-qPCR), but these methods may give false-negative results in routine samples, mainly due to low cellularity. In order to increase the sensitivity of EUS-FNA, we propose a sequential approach for detecting KRAS mutations using mutant enriched-PCR (ME-PCR, sensitivity up to 0.1%) in cytologically indeterminate and negative samples tested wild-type by RT-qPCR. EUS-FNA specimens from 107 patients with pancreatic masses (51 males, 56 females, mean age 67 years) were cytologically examined. According to the Papanicolaou Society of Cytopathology guidelines, 50 cases (47%) were classified malignant, 15 (14%) suspicious, 13 (12%) atypical and 10 (9%) negative for malignancy; 18 cases (17%) were non-diagnostic. The overall specificity and sensitivity of cytological examination were 100% and 61%, respectively, when only negative and positive cases were considered; when atypical and suspicious were added to positive cases, the sensitivity increased to 95.1% and the specificity decreased to 85.7%. In all the cases, DNA was extracted from the cell-block and KRAS mutations were investigated by RT-qPCR, followed by ME-PCR in non-amplifiable and negative cases. The overall sensitivity and specificity of KRAS mutation testing alone were 79.3% and 100%; when KRAS mutation testing was performed in indeterminate and negative cytology, the sensitivity increased to 90% with specificity to 100%. Our data indicate that conventional cytology from EUS-FNA samples is highly specific for the diagnosis of pancreatic cancer. Indeterminate and negative cases need to be screened for KRAS mutations; this two-step approach may greatly improve the diagnostic accuracy of this method.

Introduction. The area of Casale Monferrato (north west Italy) is affected by high levels of environmental asbestos pollution from an Eternit asbestos cement (AC) plant active from 1906 to 1986. We...

The aim of this study was to assess the actual effectiveness of long-term phlebotomy by comparing histological improvement (HI) in 69 Caucasian HCV-RNA-positive CHC patients undergoing phlebotomy or receiving an interferon-based therapy without virological response [nonresponders to interferon therapy(IBT-NR)].HI was defined by at least one point reduction of the staging score or, in the case of unchanged stage, by at least two points reduction of the grading score (Knodel's Activity Index) and was retrospectively evaluated by comparing two consecutive (56 ± 28 months apart) liver biopsies from 30 phlebotomized and 39 IBT-NR patients.HI was observed in 15 of 30 (50%) patients treated with phlebotomy and in six of 39 (15%) IBT-NR subjects (P=0.002). Furthermore, AST, ALT, and GGT serum levels were significantly reduced only in phlebotomized patients (P ≤ 0.003) at the time of the second biopsy. Univariate and multivariate analysis showed that histological grading score before therapy (P=0.001) and phlebotomy (P=0.002) were independently predictors of HI.HI induced by long-term phlebotomy effectively exceeds that spontaneously occurring in patients IBT-NR confirming the efficacy of iron depletion in attenuating CHC progression when other therapies have failed.

Background Evacuation disorders and hard stools are common in industrialized countries, affecting on average 12% to 17% of the adult healthy population at any age. Dietary supplementation with probiotic microorganisms may be useful in reducing the disorder. Methods We performed a double-blind, randomized, placebo-controlled study to evaluate the effectiveness of 2 different probiotic blends, either mixed Lactobacillus plantarum LP01 (LMG P-21021) and Bifidobacterium breve BR03 (DSM 16604) or Bifidobacterium animalis subspecies lactis BS01 (LMG P-21384), in the management of evacuation disorders and intestinal discomfort. In a period of 5 years (2003 to 2008), the study involved 300 healthy volunteers (151 males and 149 females; age 24 to 71 y) with evacuation disorders and hard stools. In particular, subjects were divided into 3 groups: 80 subjects in the group A received placebo, 110 subjects in the group B received mixed L. plantarum LP01 and B. breve BR03 (2.5 x 10 colony-forming units/d of each strain), and 110 subjects in the group C received B. animalis subsp. lactis BS01 (5 x 10 colony-forming units/d) for 30 days. At the beginning of the observational study, the healthy status of volunteers was evaluated by a complete, laboratory and ultrasound study of the abdomen. The physical examination was repeated after 15 and 30 days. In particular, the main troubles typically associated with evacuation disorders and hard stools as well as abdominal bloating were considered as parameters of interest. Exclusion criteria were items of gastrointestinal diseases and antibiotics intake. Results Subjects treated with the mixed probiotic strains L. plantarum LP01 and B. breve BR03 or B. animalis subsp. lactis BS01 reported a significant improvement in the number of weekly bowel movements and in the main troubles associated with evacuations, particularly consistency of feces and ease of expulsion. Discomfort items such as abdominal bloating and anal itching, burning, or pain also registered a relevant improvement in the active groups receiving probiotics. Conclusions The intake of an effective amount of mixed L. plantarum LP01 and B. breve BR03 or B. animalis subsp. lactis BS01 for 30 days is able to significantly relieve the evacuation disorders and hard stools, thus providing a useful tool for the management of such condition, which is particularly widespread in industrialized countries at any age.

Small cell lung cancer (SCLC) is a highly aggressive neoplasm that accounts for approximately 10% to 15% of lung cancers. In most cases, the diagnosis relies on cytology and needs to be confirmed by immunohistochemistry. Although several genetic and molecular abnormalities have been recorded, molecular markers able to predict the prognosis are still lacking. MicroRNA (miRNA) signatures have been recently proposed as useful biomarkers in lung cancer because of their high stability during standard sample processing.Cytological samples for 50 patients with SCLC were collected from primary tumors (n = 25) and metastases (n = 25) by means of fine-needle aspiration (FNA) or bronchial washing (BW); they were fixed in ethanol (FNA) or Duboscq-Brazil fluid (BW). The 3-miRNA panel expression (miR-192, miR-200c, and miR-205) was quantified with a TaqMan polymerase chain reaction miRNA assay and was compared with overall survival (OS) and clinicopathological data.All samples had sufficient RNA for the miRNA expression analysis to be performed, regardless of the sample source or the fixative medium. Patients with a low expression level of the 3-miRNA panel were associated with better OS in univariate (P = .032) and multivariate analyses (P = .022). Moreover, in the group of patients older than the mean age of our cohort (65.8 years), a significant OS advantage (P = .013) was seen for patients with a low expression level of the 3-miRNA panel.A specific 3-miRNA signature is potentially useful for predicting survival for patients with SCLC, and it may be feasible with cytological samples taken during standard diagnostic procedures. Cancer Cytopathol 2016;124:621-9. © 2016 American Cancer Society.

Background and Aim of the workAnkle and hindfoot injuries are common and may lead to functional impairment, disability, exclusion from occupational and daily activities. It's necessary a standardized method for assessing treatment outcomes in people with same condition and disease.American-Orthopaedics-Foot-and-Ankle-Society's-Ankle-Hindfoot-Evaluation-Scale (AOFAS-AHES) is specific to estimate clinical problems of the ankle-hindfoot.Outcome evaluation scales should be translated and culturally adapted into the language of the investigated patient.Our purpose was to translate and culturally adapt into Italian AOFAS-AHES, and to check its reproducibility and validity.MethodsAn Italian translation of the AOFAS-scale was retranslated into English by a native English and compared to the original to define a second correct Italian-version, that was submitted to 50 randomized patients operated at their ankle or hindfoot with a minimum follow-up of 6 months for cultural adaptation, and to 10 healthcare professionals to check comprehension of the medical part.To check intra and inter-observer reproducibility each patient underwent 2 interviews by interviewer-A and 1 by B. ShortForm(SF)-36-questionnaire for quality of life and Visual-Analogue-Scale (VAS) for pain were also compared for validation. The Pearson's-Correlation-Coefficient and the Intra-Class-Correlation coefficient were calculated to check inter and intra-observer reproducibility for validation.ResultsCultural adaptation revealed to be good. We obtained a good correlation of the inter and intra-observer reproducibility. Further validation of the Italian-AOFAS-AHES was obtained comparing AOFAS results to SF-36.ConclusionsItalian translation, cultural adaptation and validation of the AOFAS-AHES has been performed successfully and could be useful to improve assistance quality in care practice.

Asbestos exposure is the main risk factor for malignant pleural mesothelioma (MPM), a rare aggressive tumor. Nevertheless, on average less than 10% of subjects highly exposed to asbestos develop MPM, suggesting the possible involvement of other risk factors. To identify the genetic factors that may modulate the risk of MPM, we conducted a gene-environment interaction analysis including asbestos exposure and 15 single nucleotide polymorphisms (SNPs) previously identified through a genome-wide association study on Italian subjects. In the present study, we assessed gene-asbestos interaction on MPM risk using relative excess risk due to interaction and synergy index for additive interaction and V index for multiplicative interaction. Generalized multifactor dimensionality reduction (GMDR) analyses were also performed. Positive deviation from additivity was found for six SNPs (rs1508805, rs2501618, rs4701085, rs4290865, rs10519201, rs763271), and four of them (rs1508805, rs2501618, rs4701085, rs10519201) deviated also from multiplicative models. However, after Bonferroni correction, deviation from multiplicative model was still significant for rs1508805 and rs4701085 only. GMDR analysis showed a strong MPM risk due to asbestos exposure and suggested a possible synergistic effect between asbestos exposure and rs1508805, rs2501618 and rs5756444. Our results suggested that gene-asbestos interaction may play an additional role on MPM susceptibility, given that asbestos exposure appears as the main risk factor.

Summary Background : The role of gender in the progression of fibrosis in chronic hepatitis C is still under investigation. Aim : To investigate whether gender affects the progression of liver disease and/or hides other risk factors. Methods : A prospective series of 121 consecutive patients with chronic hepatitis C underwent liver biopsy. Grading and staging for chronic hepatitis were each evaluated according to Ishak's classification. Results : In univariate and multivariate analysis on the whole group of patients, male gender was not associated either with significant liver fibrosis (Ishak's score >2) or with cirrhosis (Ishak's score >4). On the contrary, in univariate analysis on patients aged ≤50 years, male gender was nearly significantly (P = 0.06) predictive of liver fibrosis, whereas it was not in patients >50 years. Hepatic iron grading, along with age, was an independent factor associated with fibrosis. Moreover, the values of all the variables which describe iron status were significantly higher in males aged ≤50 years in comparison with females of the same age. Conclusions : In chronic hepatitis C, male gender may be predictive of liver fibrosis only in patients aged≤50 years. Among fibrogenetic factors hidden by gender, iron status could play a major role.

OBJECTIVE To investigate the correlation between transcriptional control region (TCR) types and virus replication and the role of decreased host immunity in inducing TCR changes. STUDY DESIGN In a previous study, urine specimens from 78 unselected HIV-positive patients were independently evaluated by cytology, immunohistochemistry and nested polymerase chain reaction (n-PCR) to detect the presence of polyomaviruses. The JC virus (JCV) large T region was positive in 44/78 (56%) urine specimens by n-PCR. In the current study, these cases further underwent to n-PCR to detect TCR, and the amplified products were sequenced. The JCV types identified were compared using: (1) morphologic evidence of replication (decoy cells and/or immunohistochemical staining of cells detected using anti-SV 40 antiserum), and (2) patients' immune status (CD4+ cell counts). RESULTS TCR was successfully amplified in 30/44 cases (68%). TCR sequence analysis disclosed 6/30 archetype (20%) and 24/30 archetypelike sequences, the latter distributed as follows: 4 G2 (4/30, 13%) with G-->A substitutions in the C sequence (nt 9), and 20 CY (20/30, 67%) with A-->G substitutions in the F sequence (nt 19). There were no correlations with morphologic evidence of viral replication or immune status. CONCLUSION The present study indicated that TCR in urine samples from PML-free HIV-positive subjects are archetypes or archetypelike. Immune suppression does not seem to influence minor changes in the TCR genome, and single by mutations do not change JCV replication activity.

Background/aims In patients with chronic hepatitis C, hepatic iron concentration correlates with liver fibrosis. However, it is not clear whether this correlation merely reflects the presence of more active disease, or iron exacerbates chronic hepatitis C virus (HCV)-induced damage through activation of hepatic stellate cells and regeneration of hepatocytes. Materials and methods We studied 72 HCV-positive patients, staged according to the Ishak’s score system. We measured hepatic iron concentration with spectrophotometry and evaluated the number of hepatic stellate cells (using monoclonal antibody against alpha smooth muscle actin) and proliferating hepatocytes (using monoclonal antibody against Ki67). Iron and ferritin serum levels were also determined. Results Hepatic iron concentration correlated statistically with ferritin serum level (r = 0·59, P

Background and Objectives: In the perspective of improving accuracy of endoscopic ultrasound (EUS)-guided sampling, the efforts of innovation have focused on needle design, with the development of micro-core acquisition technology, and puncture technique variations. Available studies have shown that fine needle biopsy (FNB) needles can achieve a satisfactory diagnostic yield after fewer passes when compared to fine needle aspiration (FNA) needles, without an overall diagnostic advantage. Limited data are available to explain these results. We compared FNA and side-beveled FNB needles for micro-core/cellular material retrieval and diagnostic performance in pancreatic solid lesions. Methods: This was a single-center retrospective study of 110 consecutive patients who underwent EUS-sampling for a solid pancreatic lesion. All cellular material was collected for cell-block construction. The cytological examination was performed according to Papanicolaou. KRAS analysis was used as an ancillary technique to resolve indeterminate cases. Sensitivity and specificity were calculated in FNA (EchoTip® Ultra) and in FNB (EchoTip ProCore®) categories. All cases were evaluated for cellularity and micro-core (fragment retaining tissue architecture) retrieval. We compared FNA and FNB needles of different diameter (FNA: 19, 22, and 25G and FNB: 20, 22, and 25G), as well as large (19–20G) and small bore needles (22–25G). Results: Considering only positive cytology as diagnostic for malignancy, FNB needles showed higher sensitivity than FNA needles (86.4% vs. 54.9%, P = 0.002). If KRAS-mutated indeterminate cases were considered as malignant, overall sensitivity was 94.7%. In subcategories (FNB vs. FNA), sensitivity was 97.7% versus 92.2%, P = 0.37. Micro-cores were present in 65.2% of FNB versus 15.1% of FNA samples (P < 0.0001), rising to 95.2% in 20G-FNB category, and in 88.5% of large bore versus 20.5% of small bore needles (P < 0.0001). Considering the overall amount of cellular material (micro-cores plus cellular aggregates), FNB needles were better than FNA (91.3% vs. 62.3%, P = 0.001). The needle diameter did not influence the overall amount of cellular material retrieval since a satisfactory amount could be retrieved in both categories (large vs. small bore needles: 88.5% vs. 71.2%, P = 0.063). Conclusions: FNB needles allowed a relevant improvement in positive cytology with reduced need of KRAS mutational assessment or ancillary techniques. Dedicating all cellular material to the construction of cell-blocks and high retrieval rate of micro-cores may explain these results.

Background: Beneficial findings concerning probiotics are increasing day by day. However, one of the most important parameter which affects the probiotic activity of a microorganism is its survival during the gastroduodenal transit. Some microencapsulation techniques could be applied to bacterial cells to improve this parameter. Methods: A comparison between the intestinal colonization by microencapsulated bacteria and the same not microencapsulated strains has been conducted in a double blind, randomized, crossover study. The study (April to July 2005) involved 44 healthy volunteers. In particular, participants were divided into 2 groups: group A (21 participants) received a mix of probiotic strains Lactobacillus plantanum LP01 (LMG P-21021) and Bifidobacterium breve BR03 (DSM 16604) in an uncoated form, group B (23 participants) was given the same strains microencapsulated with a gastroresistant material. The not microencapsulated strains were administered at 5 × 10 9 colony forming units/strain/d for 21 days, whereas the microencapsulated bacteria were given at 1 × 10 9 colony forming units/strain/d for 21 days. At the end of the first period of treatment with probiotics a 3 weeks washout phase has been included in the study protocol. At the end of the washout period the groups were crossed: in detail, group A had the microencapsulated and group B the uncoated bacteria. The administered amounts of each strain were the same as the first treatment. The quantitative evaluation of intestinal colonization by strains microencapsulated or not microencapsulated was made by fecal samples examination at the beginning of the clinical trial, after 10 and 21 days of each treatment period. In particular, fecal heterofermentative Lactobacilli and Bifidobacteria have been counted. Results: A statistically significant increase in the fecal amounts of Lactobacilli and Bifidobacteria was recorded in both groups at the end of each treatment compared with do or d 42 (P < 0.0001 and P < 0.0001 at d 21 , P < 0.0001 and P < 0.0001 at d 63 for Lactobacilli and Bifidobacteria, respectively), confirming the ability of the 2 strains to colonize the human gut, either in a gastroprotected form or not. Participants treated with the microencapsulated bacteria reported a kinetics of intestinal colonization quite similar to participants who received not coated strains. Conclusions: Probiotics are able to exert many different beneficial effects on the human host. These effects are mediated by the number of viable cells which reach the gut. The microencapsulation techique used in this study is a valid strategy to significantly improve gastroresistance of strains, thus enhancing their probiotic activity and allowing the use of a 5 times lower amount.

AIM: To investigate the usefulness of mild iron depletion and the factors predictive for histological improvement following phlebotomy in Caucasians with chronic hepatitis C (CHC). METHODS: We investigated 28 CHC Caucasians with persistently elevated serum aminotransferase levels and non responders to, or unsuitable for, antiviral therapy who underwent mild iron depletion (ferritin ≤ 70 ng/mL) by long-term phlebotomy. Histological improvement, as defined by at least one point reduction in the staging score or, in case of unchanged stage, as at least two points reduction in the grading score (Knodell), was evaluated in two subsequent liver biopsies (before and at the end of phlebotomy, 48 ± 16 mo apart). RESULTS: Phlebotomy showed an excellent safety profile. Histological improvement occurred in 12/28 phlebotomized patients. Only males responded to phlebotomy. At univariate logistic analysis alcohol intake (P = 0.034), high histological grading (P = 0.01) and high hepatic iron concentration (HIC) (P = 0.04) before treatment were associated with histological improvement. Multivariate logistic analysis showed that in males high HIC was the only predictor of histological improvement following phlebotomy (OR = 1.41, 95% CI: 1.03-1.94, P = 0.031). Accordingly, 12 out of 17 (70%) patients with HIC ≥ 20 μmol/g showed histological improvements at the second biopsy. CONCLUSION: Male CHC Caucasian non-responders to antiviral therapy with low-grade iron overload can benefit from mild iron depletion by long-term phlebotomy.

Background: Iron accumulation is a well-known risk factor for the progression of chronic hepatitis C (CHC) to fibrosis. However, the profibrogenic role of the genes controlling iron homeostasis is still controversial. Aim: To evaluate the relative role of haemachromatosis (HFE), ferroportin and β-globin gene mutations in promoting iron accumulation and fibrosis in patients with CHC. Methods: Genetic analysis was performed together with the assessment of hepatic iron content and histology in 100 consecutive HIV-antibody and hepatitis B surface antigen-negative patients with biopsy-proven CHC. Results: Among the patients investigated, 12 were heterozygous for various β-globin gene mutations (39[C→T], IVS1.1[G→A], 22 7 bp deletion and IVS1.6[T→C]) and 29 carried HFE (C282Y, H63D and S65C) gene mutations. One further patient was heterozygous for both HFE (H63D) and β-globin (39[C→T]) variants, whereas 58 had the wild-type alleles of both the genes. Hepatic iron concentration (HIC) and hepatic stainable iron were significantly higher (p 2) was observed in 48 of 100 patients. Logistic regression demonstrated that age (OR 1.05; 95% CI 1.02 to 1.09; p Conclusions: Heterozygosis for β-globin mutations is a novel risk factor for both hepatic iron accumulation and the progression to fibrosis in patients with CHC.

Objective methods are needed to quantitatively assess the burden of fibrous tissue in liver biopsy specimens and its changes after treatment. The aim of this study was to assess the validity of a computer-assisted morphometric method in the evaluation of liver fibrosis in patients with chronic hepatitis C. Sixty-nine liver biopsy specimens stained with Sirius red were evaluated by two independent observers with a computer-assisted morphometric method to measure the percentage of fibrous tissue in the optic fields examined (fibrosis ratio). Furthermore, 11 pairs of liver biopsy specimens obtained before and after treatment from patients with chronic hepatitis C were evaluated with morphometry by two independent observers in order to assess in which direction fibrosis changed. In the 69 patients, the correlation of the morphometry-measured fibrosis ratio pairs by the two observers was high (r = 0.781). However, the differences between paired values were large, reaching +/- 5% in 95% of instances. The fibrosis ratios observed with morphometry by the two examiners correlated poorly with the Ishak's staging score. The two examiners agreed in 10 out of 11 instances in judging in which direction fibrosis had changed. In conclusion, using our present technique of computer-assisted morphometry, the quantitative assessment of the percentage extension of fibrous tissue was not sufficiently accurate. However, computer-assisted morphometry proved to be useful when evaluating the direction of fibrous changes in pairs of liver biopsy specimens from patients with chronic hepatitis C before and after treatment.

Background. In patients with chronic hepatitis C, the histological outcome of long term phlebotomy is unknown. Aim. To investigate biochemical and histological findings before and after phlebotomy in chronic hepatitis C. Patients. Twenty-four non-haemochromatotic patients with chronic hepatitis C were treated with long-term phlebotomy alone. Results. Hepatic iron concentration had decreased in all patients who underwent a second liver biopsy, two years after iron depletion was attained and maintained. Histological grading score decreased in four patients, was unchanged in two, and increased in five. Histological staging score decreased in two patients, was unchanged in five, and increased in four. Pretreatment high serum selenium level predicted the reduction of the inflammatory grading score in univariate analysis (p=0.008), while low serum aspartate aminotransferase (p=0.02) and low propeptide of procollagen 111 (p=0.08) levels predicted the lack of progression of liver fibrosis. Furthermore, when iron depletion was reached, significant reductions of serum levels of aminotransferase, gamma glutamyl transferase (−47%), propeptide of procollagen III, alpha foetoprotein, selenium were observed in 24 patients. No changes in serum hepatitis C virus-RNA levels were found. Conclusions. Phlebotomy alone seems to be efficacious in suppressing progression of chronic hepatitis C in some patients. Phlebotomy not only induces iron depletion, but it even modifies serum levels of other trace elements involved in the balance between oxidant and antioxidant processes.

Background and Aims: Women with chronic hepatitis C may have a slower rate of disease progression than men. We have previously demonstrated a relationship between hepatic iron concentration and liver fibrosis in patients with chronic hepatitis C. Our aim was to compare hepatic histologic findings, iron status and other factors putatively capable of determining the severity of chronic hepatitis between menstruating women and men of comparable age. Methods: We studied 21 consecutive hepatitis C virus (HCV)-RNA positive menstruating women and 24 consecutive HCV-RNA positive men of comparable age, who underwent liver biopsy for chronic hepatitis C. Alcohol intake was recorded and blood tests, HCV genotyping, serum iron, unsaturated iron binding capacity, serum ferritin, hepatic iron concentration, and liver histology were evaluated. Results: Menstruating women showed lower grading (2.7±1.5 vs 3.6±2, P=0.09) and significantly lower staging (1.38±1.11 vs 2.42±1.64, P= 0.037) scores than men of comparable age. Among the factors putatively capable of determining the severity of chronic hepatitis, only the hepatic iron concentration correlated with the hepatic histologic staging in a multivariate analysis. Iron-depleted women (transferrin saturation

In previous studies elevated Asymmetric NG, NG - dimethylarginine (ADMA) plasma levels, an endogenous nitric oxide synthase inhibitor, correlated with the severity of hepatic venous pressure gradient measurement, both in peripheral and in hepatic veins. The aim of this study was to explore whether elevated ADMA plasma levels were able to predict the presence of esophageal varices (EV) and/or large EV in patients with cirrhosis.74 cirrhotic patients who had undergone elective upper gastrointestinal endoscopy in order to assess the presence of portal hypertension and predictors of EV and/or large EV. ADMA levels were assayed by an ELISA test (Immundiagnostik AG, Germany).53 patients had EV (26/53 had large EV). Univariate analysis of low hemoglobin (p = 0.045), PT-INR (p = 0.003), albumin (p = 0.024), bilirubin (p = 0.036), Child-Pugh score (p = 0.026), and ascites (p = 0.036) predicted the presence of EV. Multivariate analysis predicted EV for only PT-INR. The presence of large EV was predicted with univariate analysis of ADMA plasma levels (p = 0.013), low hemoglobin (p0.001), PT-INR (p = 0.001), albumin (p = 0.001), bilirubin (p = 0.026), Child-Pugh score (p0.001), ascites (p = 0.004). Sensitivity, specificity, predictive positive and negative values of ADMA plasma level0.5 micromol/L(-1) in predicting large EV were 0.69 (95% CI 0.53 - 0.82), 0.51 (95% CI 0.40 - 0.62), 0.43 (95% CI 0.31 - 0.56), 0.76 (95% CI 0.62 - 0.86), while the area under the ROC curve was 0.65 (95% CI 0.51 - 0.79).ADMA plasma levels were increased in cirrhotics with more advanced liver failure but did not prove to be a useful clinical tool for predicting the presence of esophageal varices or large esophageal varices.