In vivo gene therapy of metachromatic leukodystrophy by lentiviral vectors: correction of neuropathology and protection against learning impairments in affected mice.

Metachromatic leukodystrophy (MLD) is a lipidosis caused by deficiency of arylsulfatase A (ARSA). Although the genetics of MLD are known, its pathophysiology is not understood. The disease leads to progressive demyelination and early death and no effective treatment is available. We used lentiviral vectors to deliver a functional ARSA gene (human ARSA) into the brain of adult mice with germ-line inactivation of the mouse gene encoding ARSA, As2. We report sustained expression of active enzyme throughout a large portion of the brain, with long-term protection from development of neuropathology and hippocampal-related learning impairments. We show that selective degeneration of hippocampal neurons is a central step in disease pathogenesis, and provide evidence that in vivo transfer of ARSA by lentiviral vectors reverts the disease phenotype in all investigated areas. Therefore, in vivo gene therapy offers a unique option for MLD and other storage diseases affecting the central nervous system.

Mother to child transmission of hepatitis C virus in a province of northern Italy.

INTRODUCTION: Study reports of mother to child transmission of hepatitis C virus (HCV) have shown transmission rates ranging from 3 to 37%, according to maternal viremia and HIV-1 coinfection. The present study evaluated the prevalence of the HCV infection in the general population and the incidence of vertical transmission, from women who delivered in the Obstetric Clinic of the Hospital of Parma from January 1st 1996 to 31st 2001 December. METHODS: Mothers and children were tested for the presence of HCV-RNA within one week after delivery. Children were considered to be infected when they were found positive at least twice for viral RNA or antibodies were still detectable at the end of the follow-up period (18 months) in blood. RESULTS: Out of 13,025 women, 110 (0.8%) were found positive for anti-HCV antibodies; 72 of them (65.4%) were HCV-RNA positive. All 110 children were positive for anti-HCV antibodies in the first blood sample (time 0); 8 of them were HCV-RNA positive. Three children were still viremic at the end of the follow-up whereas 5 showed a clearance. No significant differences were found between viremic and nonviremic children with respect to gestational week, maternal alanine aminotransferase (ALT) levels and newborns weight at birth. CONCLUSION: This investigation shows that vertical transmission may occur in a general obstetric population despite a low prevalence of HCV-positive subjects.

Ganglioside GM2 is substrate for a sialidase in MDCK cells.

GM1 ganglioside carrying a fluorescent fatty acid in substitution of the natural one, has been administered to cultured Madin-Darby canine kidney (MDCK) cells for different pulse times (0.5-24 h), and its metabolic fate was followed. The fluorescent GM2, asialo-GM2, asialo-GM1 and ceramide were the only detectable metabolites. The complete absence of fluorescent GM3 is consistent with the presence in these cells of a sialidase working on GM1 and GM2 gangliosides. After treatment of the cells with chloroquine the fluorescent GM1 remained essentially undegraded, indicating a catabolic processing at lysosomal level.

Nerve growth factor induces sphingomyelin accumulation in pheochromocytoma cells.

The pheochromocytoma cells are a well-known model for studying the nerve growth factor (NGF)-induced molecular changes during the differentiation process. The involvement of sphingomyelin (SM) was studied using the fluorescent analogue of ceramide, i.e. N-lissamine rhodaminyl-(12-aminododecanoyl) D-erythro-sphingosine (C12-LRh-Cer). This fluorescent analogue is metabolically active and can be used to follow the biosynthesis of SM in intact cells. NGF induces a 4-fold increase of fluorescent SM content in PC12 cells, when loaded with C12-LRh-Cer. Treatment of PC12 cells with actinomycin D or cycloheximide completely abolishes the NGF-induced elevation of SM. Inhibition of p140(trkA) receptor by AG-879 prevents extracellular signal-regulated kinase 1/2 phosphorylation and suppresses the increase of SM. Inhibition of protein kinase C (PKC), protein kinase A (PKA) and phosphatidylinositol 3-kinase does not have any effect on NGF-induced C12-LRh-SM accumulation. On the other hand, activation of PKA or PKC with simultaneous treatment with NGF has a synergistic effect on increase of SM content. The NGF-induced SM increase in PC12 cells is an effect promoted by other differentiating agents like dibutyryl cyclic AMP or fibroblast growth factor-2 but not by a mitogenic agent like epidermal growth factor.

[Considerations on the complications of epidemic parotitis]. / Considerazioni sulle complicanze di un'epidemia di parotite.

The AA describe 82 cases of complications occurred during an outbreak of mumps in 1980-81. Forty-two cases of meningitis, 4 of pancreatitis, 2 of orchitis and 2 cases of encefalitis occurred. All patients had eventual, complete recovery. The other cases are not specific complications. The treatment was almost entirely symptomatic. The headache of meningitis was relieved by a lumbar poncture. In the epicrisis of the AA these cases of complications have some importance in relation to the opportunity of getting herd vaccination against mumps.

[The pediatrician and the child in a multiethnic society]. / Il pediatra ed il bambino nella società multietnica.

The increasing immigration from developing countries has determined new health problems. Italian Pediatricians must become acquainted with these health concerns such as the excessive use of public pediatric structures, especially hospitals, specific diseases unusual among Italian children (malaria, tuberculosis, tropical parassitosis, sickle cell anemia, rickets and others), control of previous immunizations and repetition when one is not sure of their efficacy. Moreover, immigrated children seem to be more susceptible to common pediatric diseases (mostly infections) than Italian children. Finally, children adopted from developing countries deserve special attention because they arrive from a very poor social context with frequent history of important infectious diseases, congenital diseases, malnutrition and abuse. Central precocious puberty is a frequent evolution in some of the adopted girls.

Poliomyelitis immunity status at different intervals from vaccination.

Research on neutralizing antibodies against the 3 serotypes of poliovirus was carried out on 441 blood specimens drawn from children and young people living in Parma, a town of about 200,000 inhabitants, where the vaccination services were well organized. From June 1983 to June 1984 blood samples were taken from different groups of subjects at different stages of vaccination (before vaccination, and after 1, 2, 3, and 4 doses of trivalent balanced OPV). In addition, groups of children and young people vaccinated from 2 to more than 14 years earlier were bled. Ten per cent of unvaccinated babies (aged 1-3 months) were lacking in neutralizing antibodies for types 1 and 3 and five per cent for type 2; among children aged 4-5 months vaccinated by one dose of OPV a residual 5% of subjects without antibodies for type 1 was also found. All subjects who received two, three, or four doses, had antibodies against all serotypes of poliovirus. As for the number of administered doses, significant differences among GMTs of antibodies were recorded only for type 1, when the group given 3 doses, was compared with the group which received two and one dose, respectively. At different intervals from completion of vaccination (2-14 years), all groups we examined still had antibodies against all three serotypes. For poliovirus type 1 antibody, GMTs were not significantly different in the examined groups. For types 2 and 3 significant decreases were observed in the groups vaccinated from 2 to 5 years. The high levels of immunity observed earlier in Parma were not found throughout the entire territory.(ABSTRACT TRUNCATED AT 250 WORDS)

[Vertical transmission of HIV infection: risk factors and prevention strategies]. / Trasmissione verticale dell'infezione da HIV: fattori di rischio e strategie di prevenzione.

In this study it has been evaluated the connection between the maternal immune status and the obstetrical factors on the one hand end the perinatal transmission of HIV in the other as well as, the effects of zidovudine prophylaxis and elective caesarean section on vertical HIV transmission. From January 1987 to September 1999, 60 HIV infected pregnant women were followed with their 64 infants. From July 1995 15 women and their respective children were treated with zidovudine prophylaxis (protocol ACTG 076); from June 1997, in 8 of these patients a systematic caesarean section was performed (7 electively and 1 on an emergent basis). The transmission rare was 20.5% among the 44 children in the group without zidovudine prophylaxis, compared with 7.1% among the 14 children in the group receiving Zidovudine. As for as the mother-child pairs receiving Zidovudine therapy, the rate of vertical transmission was 0% for the 7 mothers who underwent elective cesarean section and 14.3% for the 7 mothers with other ways of delivery. The interaction between zidovudine prophylaxis and elective caesarean section was associated with the lowest rate of vertical HIV transmission.

[Acute respiratory syndromes caused by viruses in a pediatric department. Clinical and epidemiological studies of a series of cases admitted in the period from February to May, 1979]. / Le sindromi respiratorie acute da virus in un reparto pediatrico. Studi clinico-epidemiologici su una casistica raccolta nel periodo febbraio-maggio 1979.

In the period between February and May 1979, in concomitance with a sharp increase in admittance to the pediatric Division of the Parma General Hospital for acute respiratory infections, we carried out a virologic and serologic investigation on hospitalized children aged 21 days-12 years. The investigation was carried out on 137 patients with respiratory diseases: 33 with bronchiolitis, which occurred mainly (32/33) between February and March in children that were prevalently (30/33) in the first year of life; 27 with upper respiratory tract infections; 54 with mid respiratory tract infections; 23 with pneumonia. Forty-eight controls (without respiratory infections) were also investigated. In the months of February and March there was a high circulation of respiratory syncytial virus (RSV): throughout the entire period there was a lower incidence of Adenovirus infections and more uniformly distributed.

[Cholestasis caused by ulcerative Vater's papillitis in a child with AIDS]. / Cholestase secondaire à une papillite ulcérative de Vater chez un enfant atteint de SIDA.

A case of cholestasis induced by ulcerative papillitis in an 11 month-old boy with AIDS is described. Resolution of cholestasis has been achieved with medical therapy. The authors underline the necessity of repeated endoscopic follow-up to evaluate the risk of secondary papillary stenosis, a complication usually observed in adults.

[Vertical transmission of the hepatitis C virus]. / La trasmissione verticale del virus dell'epatite C.

The incidence of hepatitis C in childhood is approximately 0.4%. The mode of transmission can be parenteral, sexual, occupational and also vertical. The latter has an incidence that varies widely and it increases in the case of human immuno-deficiency virus (HIV) coinfection and high titers of HCV in the mother. The vertical transmission is not influenced by breast feeding, however, data are discrepant with regard to child delivery (cesarean section vs vaginal delivery). Ninety-seven babies born from mothers with hepatitis C from 1996 to 1999, were evaluated prospectively in the Day Hospital of the Pediatric Department of Parma. The protocol of observation established a blood sampling for titers of antibodies anti-HCV and HCV-RNA at the 3rd trimester of pregnancy and subsequent clinical and biochemical controls at 3-6-9-12-15 and 18 months. Thirty (31.2%) out of the 96 mothers evaluated were positive for antibodies anti-HCV and 66 (68.8%) were positive for antibodies anti-HCV and HCV-RNA. Five (5.15%) out of the 97 babies evaluated were infected by HCV. Of these 4 were delivered vaginally and 1 by cesarean section. Of the 3 babies born to mothers with HIV coinfection, none was infected by HIV, but 1 was infected by HCV. Vertical transmission is increased by HCV viral load or HIV coinfection in the mother. The vaginal delivery and breastfeeding do not represent an additional risk factor.

The prevalence of HCV infection in a cohort of pregnant women, the related risk factors and the possibility of vertical transmission.

The prevalence of antibodies for one or more HCV antigens was 2.3% of 1,347 mothers at childbirth. Compared with the principal factors studied, the presence of antibodies was more frequent in women who were carriers of HIV infection (3/3), in those who had suffered liver diseases (5/37) or who had had transfusion (3/25). This was as opposed to women who did not have any risk factor (p < 0.001). The prevalence of HCV-RNA was 1.3%; in relation to the antibody state, such a condition was more frequent in subjects with antibodies for 3 or 4 antigens (about 80%) compared with those who were positive for 1 or 2 antigens. HCV-RNA of the same genotype as the mother (type 1; 1a) was also found in the funicular blood of 2 of the 18 babies born to mothers who were positive for HCV-RNA. In the course of the follow-up (from the 3rd to the 18th month) the viral RNA was not found in any of the babies, nor was it found in the 2 who were positive at birth. Even the antibodies gradually disappeared, although slowly. At the 10th month, 91% of the babies resulted as having no antibodies and at the 18th month none of the babies resulted as having antibodies. Breast-feeding also appeared to have no influence on the transmission of the infection; out of 18 viremic mothers indeed 12 (67%) breast-fed their babies.