- Email: [email protected]
[680] MUTATIONS OF ABCB4 GENE IN CHILDREN WITH PROGRESSIVE FAMILIAL INTRAHEPATIC CHOLESTASIS: A MULTICENTER ITALIAN STUDY
07. PEDIATRIC HEPATOLOGY
-
during maintenance venesection. Our aim was to assess if the venesection programme in our unit achieves target ferritin and transferrin saturation levels in patients with haemochromatosis. Prior to publication of guidelines our unit had been aiming for a serum ferritin of 100ng/ml and transferrin saturation of 4 0 % . Methods: Data was collected from charts and computerised laboratory records. Ferritin and transferrin levels at end of initial regular venesection phase (IRVP) and during maintenance venesection (MV) were recorded. In addition, alcohol consumption was noted. Results: Charts were available on 99 patients (73 male). Fifty five patients had completed the TRVP at our unit. Forty of the 55 (72.7%) achieved a serum ferritin of <100ng/ml and 37 (67.3%) had a transferrin saturation under 50% at the end of TRVP. During the MV phase, 80% patients had a recorded ferritin < I OOngiml and 59% achieved a transferrin saturation under 50%. However 33 of 99 patients (33.3%) were no longer under review, 21 of whom had failed to attend (FTA), 7 were deceased, 2 transplanted, 3 other. When FTAs are included, only 55% of those who should be attending for MV achieved our target ferritin. 34% of patients exceeded the recommended weekly maximum alcohol consumption. Conclusions: Our unit currently achieves the target serum ferritin level in 72.7% at end of TRVP and 80.43% in those patients who attend for maintenance venesection. There is, however, a high rate offailure to attend for maintenance venesection and a high level of excess alcohol intake. A haemochromatosis treatment co-ordinator may improve both these figures.
16801 MUTATIONS OF ABCB4 GENE IN CHILDREN WITH PROGRESSIVE FAMILIAL INTRAHEPATIC CHOLESTASIS: A MULTICENTER ITALIAN STUDY C. Colombo’, P. Vajro’, R. Torio’, E. Castellano3, M.G. Marazzi3, S. Martelossi4, L. Costantino’, D. De Giorgio’, G. Maggiore’. ‘Department of Pediutricc., Uniuer~c.it?,of Milun, Fondazione IRCCS Os~~edale MuggiorePoliclinico, Milano; 2Dipartimento di Pediutriu, linioersitd di Napoli Federico 11, Napoli; Dipartimento di Pediatia, IRCCS lstituto G. Gaslini, Genoua; 4Dipartinzento di Pediatria, IRCCS Burlo Garofdo, Trieste; iDipartinzento di Pediatria, linioersitd di Pisa, Pisa, Italy E-mail: [email protected]
’
PFlC is an heterogeneous clinical entity characterized by a progressive clinical course during infancy and eventually end-stage liver disease. Evidence has been provided that PFlC type 3 is caused by mutations of ABCB4 gene which encodes a class 111multidrug resistance P-glycoprotein involved in biliary secretion of phospholipids. Our aim was to assess frequency of ABCB4 mutations among patients with PFlC and high gammaGT activity enrolled through a multicenter study within the Italian Society for Pediatric Gastroenterology. Information of patients included demographics, family history and major clinical events, biochemical parameters, imaging and histological findings. Genetic analysis of ABCB4 mutations was performed in 72 PFlC children and in 50 healthy subjects. Molecular analysis was performed by DNA amplification of the 27 exons of ABCB4 gene, analysis of PCR products by DHPLC; fragments with abnormal size were then sequenced. Ofthe 72 enrolled patients, 19 carried 38 ABCB4 gene mutations, that were found in none of the control subjects; there were 28 different mutations located mostly in the central region of the protein and 24 are novel (18 missense and 6 produce a truncated protein). Comparison of 18 aminoacidic loci of the 18 novel missense mutations with the corresponding loci in different species showed that these mutations are located in a very conserved position. Eleven of the 19 affected patients were males, age at onset was 10.5 months, ranging from birth to 16 yrs. Positive family history for intrahepatic cholestasis of pregnancy was present in 4 mothers; early complicated biliary lithiasis in 3 parents and drug-induced cholestasis in one mother. Parents were consanguineous in 2 cases. Symptoms at onset included pruritus in 12 and jaundice in 3. Follow-up ranged from 10 months to 21 yrs. None of
INHERITED DISEASES OF THE LIVER
S257
the patients died, one was transplanted, 6 developed portal hypertension. Ursodeoxycholic acid treatment was associated with normalisation or improvement of liver enzymes in I 1 patient. In conclusion, a wide spectrum of ABCB4 mutations is associated with PFIC; most are located in 3 exons; ABCB4 mutations accounts only for a limited number of patients with PFlC and high gammaGT activity.
16811 STAGING OF FIBROSIS BY LIVER STIFFNESS MEASUREMENT IN THALASSEMICS WITH IRON OVERLOAD
V Di Marco’, F. Bronte’, D. Cabibi’, F, Barbaria’, Z. Borsellino3, G. Alaimo’, S. Ciminnisi’, M. Capra3, A. Maggio4, P.L. Almasio’, und Hepatology; Unit ofHistology, A. Craxi’ . ‘Unit (?fGu~stroenterolog~~ linioersiiy of‘ Palernzo; .’linit of‘ Thalassenzia, Ospedale G. Di Cristina, Pulermo; Unit of Thalassemiu, Ospedule ci Ceruello, Pulermo, ltuly E-mail: [email protected] Background and Aims: Several studies report a close relation between liver stiffness measured by FibroScan (Echosens, France) and fibrosis evaluated by liver biopsy (LB), but the influence of iron overload on liver stiffness measurement (LSM) is unknown. Aim of o w study was to evaluate the correlation between liver fibrosis, iron overload and LSM in patients with iron overload due to thalassemia major. Methods: Forty-seven patients with homozygous fl thalassemia (1 1 children, mean age 9 years; 36 adults, mean age 32 years and mean BMI 24.4kg/m2; 19 HCV-RNA positive and 28 anti-HCV and HCV-RNA negative) and 41 non-thalassemic adults with chronic HCV hepatitis (all HCV-RNA positive, mean age 55 years, mean BMT 27.7kg/m2) underwent LB (mean length of biopsy 15 mm) and simultaneous LSM. Liver inflammation and fibrosis were scored according to METAVTR, and LTC measured on fresh tissue cores weighing more than 4 mg by atomic absorption spectrometry. Results: The degree of liver fibrosis, the LSM expressed as M a , and the LIC expressed as mg/gr of liver dry weight in thalassemics and non thalassemic patients. Among thalassemics, values of LSM and LTC were not significantly different for each class of fibrosis according to the HCV infection status. LSM increased proportionally to the METAVIR stage, with a highly significant relation to fibrosis (P > 0.001 by ANOVA test). There was no obvious relation between LTC and LSM even at the highest degrees of liver siderosis. LSM was had excellent value in discriminating thalassemia patients with F4 fibrosis (i.e. cirrhosis), but was less performing at lower stages of fibrosis. Conclusion: LSM by FibroScan is an adequate and reliable non-invasive tool to diagnose advanced liver fibrosis in subjects with severe liver iron overload
16821 INTRAUTERINE-TRANSPLACENTAL TRANSMISSION OF HEPATITIS B VIRUS (HBV) INFECTION FROM VlREMlC (PRECORE MUTANT, G1896A) CHRONIC HBV INFECTED MOTHERS TO THEIR INFANTS T.S. Elefsiniotis’, M. Papadalcis’, G. Vlachos’, G. Daskalakis’ , C. Barbatis3, A. Antsaklis’ . ‘First Department of Obstetric and Gynecology, Alexandras liniuersiw Hospital, Athens; 2 D e p m n e n t of Internal Medicine, Hippokration Hospitul, Athens; ‘Department of Puthology, Red Cross Hospital, Athens, Greece E-mail: [email protected] Background and Aim: Tntrauterine/transplacental transmission of HBV infection in infants born from HBsAg(+) mothers is positively related to HBeAg(+) maternal serological status and high levels of maternal serum HBV-DNA. Despite the predominance of HBeAg(-) serological status ( ~ 9 5 % among ) HBsAg(+) pregnant women in Greece about a third
-
during maintenance venesection. Our aim was to assess if the venesection programme in our unit achieves target ferritin and transferrin saturation levels in patients with haemochromatosis. Prior to publication of guidelines our unit had been aiming for a serum ferritin of 100ng/ml and transferrin saturation of 4 0 % . Methods: Data was collected from charts and computerised laboratory records. Ferritin and transferrin levels at end of initial regular venesection phase (IRVP) and during maintenance venesection (MV) were recorded. In addition, alcohol consumption was noted. Results: Charts were available on 99 patients (73 male). Fifty five patients had completed the TRVP at our unit. Forty of the 55 (72.7%) achieved a serum ferritin of <100ng/ml and 37 (67.3%) had a transferrin saturation under 50% at the end of TRVP. During the MV phase, 80% patients had a recorded ferritin < I OOngiml and 59% achieved a transferrin saturation under 50%. However 33 of 99 patients (33.3%) were no longer under review, 21 of whom had failed to attend (FTA), 7 were deceased, 2 transplanted, 3 other. When FTAs are included, only 55% of those who should be attending for MV achieved our target ferritin. 34% of patients exceeded the recommended weekly maximum alcohol consumption. Conclusions: Our unit currently achieves the target serum ferritin level in 72.7% at end of TRVP and 80.43% in those patients who attend for maintenance venesection. There is, however, a high rate offailure to attend for maintenance venesection and a high level of excess alcohol intake. A haemochromatosis treatment co-ordinator may improve both these figures.
16801 MUTATIONS OF ABCB4 GENE IN CHILDREN WITH PROGRESSIVE FAMILIAL INTRAHEPATIC CHOLESTASIS: A MULTICENTER ITALIAN STUDY C. Colombo’, P. Vajro’, R. Torio’, E. Castellano3, M.G. Marazzi3, S. Martelossi4, L. Costantino’, D. De Giorgio’, G. Maggiore’. ‘Department of Pediutricc., Uniuer~c.it?,of Milun, Fondazione IRCCS Os~~edale MuggiorePoliclinico, Milano; 2Dipartimento di Pediutriu, linioersitd di Napoli Federico 11, Napoli; Dipartimento di Pediatia, IRCCS lstituto G. Gaslini, Genoua; 4Dipartinzento di Pediatria, IRCCS Burlo Garofdo, Trieste; iDipartinzento di Pediatria, linioersitd di Pisa, Pisa, Italy E-mail: [email protected]
’
PFlC is an heterogeneous clinical entity characterized by a progressive clinical course during infancy and eventually end-stage liver disease. Evidence has been provided that PFlC type 3 is caused by mutations of ABCB4 gene which encodes a class 111multidrug resistance P-glycoprotein involved in biliary secretion of phospholipids. Our aim was to assess frequency of ABCB4 mutations among patients with PFlC and high gammaGT activity enrolled through a multicenter study within the Italian Society for Pediatric Gastroenterology. Information of patients included demographics, family history and major clinical events, biochemical parameters, imaging and histological findings. Genetic analysis of ABCB4 mutations was performed in 72 PFlC children and in 50 healthy subjects. Molecular analysis was performed by DNA amplification of the 27 exons of ABCB4 gene, analysis of PCR products by DHPLC; fragments with abnormal size were then sequenced. Ofthe 72 enrolled patients, 19 carried 38 ABCB4 gene mutations, that were found in none of the control subjects; there were 28 different mutations located mostly in the central region of the protein and 24 are novel (18 missense and 6 produce a truncated protein). Comparison of 18 aminoacidic loci of the 18 novel missense mutations with the corresponding loci in different species showed that these mutations are located in a very conserved position. Eleven of the 19 affected patients were males, age at onset was 10.5 months, ranging from birth to 16 yrs. Positive family history for intrahepatic cholestasis of pregnancy was present in 4 mothers; early complicated biliary lithiasis in 3 parents and drug-induced cholestasis in one mother. Parents were consanguineous in 2 cases. Symptoms at onset included pruritus in 12 and jaundice in 3. Follow-up ranged from 10 months to 21 yrs. None of
INHERITED DISEASES OF THE LIVER
S257
the patients died, one was transplanted, 6 developed portal hypertension. Ursodeoxycholic acid treatment was associated with normalisation or improvement of liver enzymes in I 1 patient. In conclusion, a wide spectrum of ABCB4 mutations is associated with PFIC; most are located in 3 exons; ABCB4 mutations accounts only for a limited number of patients with PFlC and high gammaGT activity.
16811 STAGING OF FIBROSIS BY LIVER STIFFNESS MEASUREMENT IN THALASSEMICS WITH IRON OVERLOAD
V Di Marco’, F. Bronte’, D. Cabibi’, F, Barbaria’, Z. Borsellino3, G. Alaimo’, S. Ciminnisi’, M. Capra3, A. Maggio4, P.L. Almasio’, und Hepatology; Unit ofHistology, A. Craxi’ . ‘Unit (?fGu~stroenterolog~~ linioersiiy of‘ Palernzo; .’linit of‘ Thalassenzia, Ospedale G. Di Cristina, Pulermo; Unit of Thalassemiu, Ospedule ci Ceruello, Pulermo, ltuly E-mail: [email protected] Background and Aims: Several studies report a close relation between liver stiffness measured by FibroScan (Echosens, France) and fibrosis evaluated by liver biopsy (LB), but the influence of iron overload on liver stiffness measurement (LSM) is unknown. Aim of o w study was to evaluate the correlation between liver fibrosis, iron overload and LSM in patients with iron overload due to thalassemia major. Methods: Forty-seven patients with homozygous fl thalassemia (1 1 children, mean age 9 years; 36 adults, mean age 32 years and mean BMI 24.4kg/m2; 19 HCV-RNA positive and 28 anti-HCV and HCV-RNA negative) and 41 non-thalassemic adults with chronic HCV hepatitis (all HCV-RNA positive, mean age 55 years, mean BMT 27.7kg/m2) underwent LB (mean length of biopsy 15 mm) and simultaneous LSM. Liver inflammation and fibrosis were scored according to METAVTR, and LTC measured on fresh tissue cores weighing more than 4 mg by atomic absorption spectrometry. Results: The degree of liver fibrosis, the LSM expressed as M a , and the LIC expressed as mg/gr of liver dry weight in thalassemics and non thalassemic patients. Among thalassemics, values of LSM and LTC were not significantly different for each class of fibrosis according to the HCV infection status. LSM increased proportionally to the METAVIR stage, with a highly significant relation to fibrosis (P > 0.001 by ANOVA test). There was no obvious relation between LTC and LSM even at the highest degrees of liver siderosis. LSM was had excellent value in discriminating thalassemia patients with F4 fibrosis (i.e. cirrhosis), but was less performing at lower stages of fibrosis. Conclusion: LSM by FibroScan is an adequate and reliable non-invasive tool to diagnose advanced liver fibrosis in subjects with severe liver iron overload
16821 INTRAUTERINE-TRANSPLACENTAL TRANSMISSION OF HEPATITIS B VIRUS (HBV) INFECTION FROM VlREMlC (PRECORE MUTANT, G1896A) CHRONIC HBV INFECTED MOTHERS TO THEIR INFANTS T.S. Elefsiniotis’, M. Papadalcis’, G. Vlachos’, G. Daskalakis’ , C. Barbatis3, A. Antsaklis’ . ‘First Department of Obstetric and Gynecology, Alexandras liniuersiw Hospital, Athens; 2 D e p m n e n t of Internal Medicine, Hippokration Hospitul, Athens; ‘Department of Puthology, Red Cross Hospital, Athens, Greece E-mail: [email protected] Background and Aim: Tntrauterine/transplacental transmission of HBV infection in infants born from HBsAg(+) mothers is positively related to HBeAg(+) maternal serological status and high levels of maternal serum HBV-DNA. Despite the predominance of HBeAg(-) serological status ( ~ 9 5 % among ) HBsAg(+) pregnant women in Greece about a third