The Methylation Analysis of the Glucose-Dependent Insulinotropic Polypeptide Receptor (GIPR) Locus in GH-Secreting Pituitary Adenomas.

: The glucose-dependent insulinotropic polypeptide receptor (GIPR) is aberrantly expressed in about one-third of GH-secreting pituitary adenomas (GH-PAs) and has been associated with a paradoxical increase of GH after a glucose load. The reason for such an overexpression has not yet been clarified. In this work, we aimed to evaluate whether locus-specific changes in DNA methylation patterns could contribute to this phenomenon. By cloning bisulfite-sequencing PCR, we compared the methylation pattern of the GIPR locus in GIPR-positive (GIPR+) and GIPR-negative (GIPR-) GH-PAs. Then, to assess the correlation between Gipr expression and locus methylation, we induced global DNA methylation changes by treating the lactosomatotroph GH3 cells with 5-aza-2'-deoxycytidine. Differences in methylation levels were observed between GIPR+ and GIPR- GH-PAs, both within the promoter (31.9% vs. 68.2%, p < 0.05) and at two gene body regions (GB_1 20.7% vs. 9.1%; GB_2 51.2% vs. 65.8%, p < 0.05). GH3 cells treated with 5-aza-2'-deoxycytidine showed a ~75% reduction in Gipr steady-state level, possibly associated with the observed decrease in CpGs methylation. These results indicate that epigenetic regulation affects GIPR expression in GH-PAs, even though this possibly represents only a part of a much more complex regulatory mechanism

Determinants of mother-to-infant human immunodeficiency virus 1 transmission before and after the introduction of zidovudine prophylaxis

Background: Randomized controlled trials have demonstrated that zidovudine therapy decreases the mother-to-infant transmission of human immunodeficiency virus 1 (HIV-1). Data from large observational studies may provide further important findings on the effectiveness at the population level of combined treatments in decreasing transmission. Objective: To evaluate time trends in prophylactic interventions and the determinants of transmission both before and after the introduction of antiretroviral prophylaxis, and in treated and untreated mother-infant pairs after 1995. Design and Setting: Analysis of prospective data on 3770 children born to HIV-1-infected women between 1985 and 1999 and reported to the Italian Register for HIV Infection in Children. Main Outcome Measures: Logistic regression random effects models were used to estimate crude and adjusted odds ratios for several factors potentially influencing vertical transmission for 2periods-1985 through 1995 (January 1, 1985, through December 31,1995) and 1996 through 1999 (January 1, 1996, through December 31, 1999), and between treated and untreated children after 1995. Results: The transmission rate was 15.5% in the 19851995 period and 5.8% in the 1996-1999 period. By 1999, prophylactic interventions had greatly increased. Antiretroviral treatment (ART) usage was 89.9%, (55.1% combination ART) and the elective cesarean delivery rate was 81.3%. In multivariate analysis, only elective cesarean delivery was associated with a lower risk of mother-to-infant transmission before 1995. After 1995, nonbreastfeeding and receipt of ART were protective whereas elective cesarean delivery was not significantly protective in multivariate analysis. Transmission risk was reduced by 76% with an incomplete zidovudine regimen, 88% with a complete regimen, and 93% when the mother received combination ART. In the 1996-1999 period, the transmission rate for nonbreastfeeding mother-infant pairs was 8.6% with elective cesarean delivery, 4.4% with any ART, and 2.4% with these interventions combined

Persistently high IgA serum levels are a marker of immunological or virological failure of combined antiretroviral therapy in children with perinatal HIV-1 infection

Non-expensive and low-complexity surrogate markers for monitoring the response to combined antiretroviral therapy (combined-ART) are needed in poor-resource settings where routine assessment of CD4+ T-lymphocyte count and viral load can not be afforded. We longitudinally evaluated Ig serum levels in 234 HIV-1 infected children receiving combined-ART with ≥ 3 drugs. Since Ig levels physiologically vary with age, differences at different age periods were evaluated as differences in z-scores calculated using the mean and standard deviation of the normal population for each age period. Data from 17 (7·3%) children with immunological failure and from 54 (23·1%) children with virological failure of combined-ART were compared with data from not-failed children. At baseline children with immunological failure showed higher IgM z-scores (P = 0·042) than children without. After 3–12 months of therapy immunologically failed children displayed higher viral loads (P < 0·0001) and IgA (P = 0·043) z-scores than not-failed children. Similarly, at the same follow-up time, children with virological failure showed lower CD4(+) T-lymphocyte percentages (P = 0·005) and higher IgA z-scores (P < 0·0001) than not-failed children. No difference in IgG or IgM z-scores was evidenced between failed and not-failed children after 3–12 months of therapy. In conclusion, IgA serum level is a cheap and low-complexity marker of immunological or virological failure of combined-ART which might be adopted in poor-resource settings

Combined antiretroviral therapy reduces hyperimmunoglobulinemia in HIV-1 infected children

Objective: To evaluate the effect of combined antiretroviral therapy on serum immunoglobulin (Ig) levels in HIV-1 perinatally infected children. Methods: Data from 1250 children recorded by the Italian Register for HIV Infection in Children from 1985 to 2002 were analysed. Since Ig levels physiologically vary with age, differences at different age periods were evaluated as differences in z-scores calculated using means and standard deviations of normal population for each age period. Combined antiretroviral therapy has become widespread in Italy since 1996, thus differences in Ig z-scores between the periods 1985-1995 and 1996-2002 were analysed. Data according to type of therapeutic regimen were also analysed. Results: Between the two periods 1985-1995 and 1996-2002, significant (P < 0.0001) decreases in IgG (6.29 +/- 4.72 versus 4.44 +/- 4.33), IgM (9.25 +/- 13.32 versus 5.61 +/- 7.93), and IgA (10.25 +/- 15.68 versus 6.48 +/- 11.56) z-scores, together with a parallel significant (P < 0.0001) increase in CD4 T-lymphocyte percentages, were found. These decreases were confirmed regardless of whether the children were receiving intravenous Ig or not. Ig z-scores were significantly higher in children receiving mono-therapy than in those receiving double-combined therapy (IgG, P < 0.0001; IgM, P = 0.003; IgA, P = 0.031) and in the latter children than in those receiving three or more drugs (P < 0.0001 for all z-scores). Ig z-scores correlated inversely with CD4 T lymphocyte percentages and, directly, with viral loads. Conclusions: Our data show that in HIV-1 infected children combined antiretroviral therapy leads to reduction of hyperimmunoglobulinemia which parallels restoration of CD4 T-lymphocyte percentage and viral load decrease, which it turn probably reflects improved B-lymphocyte functions

Lower mother to child HIV-1 transmission in boys is independent of type of delivery and antiretroviralprophylaxis

The relationship between infant's gender and rate of HIV-1 mother-to-child transmission (MTCT) was evaluated in a prospective cohort of 4151 children (2166 boys and 1985 girls) born to HIV-1-infected mothers enrolled in the Italian Register for HIV Infection in Children. Logistic regression models were performed to estimate crude odds ratios (ORs) and adjusted odds ratios (AORs) and 95% CIs for factors potentially influencing MTCT separately for the period 1985-1995 and the period 1996-2001. To evaluate rates of MTCT by gender in specific subgroups, separate logistic regression models by mode of delivery and antiretroviral prophylaxis were performed. Among children born in 1985-1995, 15.5% boys (95% Cl: 13.6-17.7) and 17.9% girls (95% Cl: 15.7-20.3) were infected (P = 0.1181). After 1995, a lower proportion of boys (3.1% [95% CI: 2.0-4.4]; AOR: 0.43 [95% Cl: 0.26-0.71], P = 0.0008) than girls (AOR: 6.3%, 95% CI: 4.8-8.1) was infected. Lower AORs for boys persisted independently of elective cesarean delivery (AOR: 0.31, 95% Cl: 0.14-0.71); other than elective cesarean (AOR: 0.38, 95% Cl: 0.19-0.78) and antiretroviral prophylaxis (zidovudine monotherapy (AOR: 0.11, 95% CI: 0.03-0.38); none (AOR: 0.43, 95% CI: 0.21-0.90). No difference was observed when combined therapy in the mother was administered (AOR: 1.14, 95% CI: 0.30-4.32), but results were likely to be biased by the very low rate of infected children in this group. A lower proportion of HIV-1-infected boys in children born after 1995 was found. Factor(s) intrinsic to gender (rather than type of delivery or maternal antiretroviral prophylaxis) may be involved, because the risk of infection in boys was lower independent of interventions. A possible explanation is that, among infected fetuses, more girls survive up to the end of pregnancy and may take advantage of the benefits of preventive strategies

Determinants of mother-to-infant human immunodeficiency virus 1 transmission before and after the introduction of zidovudine prophylaxis

Background: Randomized controlled trials have demonstrated that zidovudine therapy decreases the mother-to-infant transmission of human immunodeficiency virus 1 (HIV-1). Data from large observational studies may provide further important findings on the effectiveness at the population level of combined treatments in decreasing transmission.Objective: To evaluate time trends in prophylactic interventions and the determinants of transmission both before and after the introduction of antiretroviral prophylaxis, and in treated and untreated mother-infant pairs after 1995.Design and Setting: Analysis of prospective data on 3770 children born to HIV-1-infected women between 1985 and 1999 and reported to the Italian Register for HIV Infection in Children.Main Outcome Measures: Logistic regression random effects models were used to estimate crude and adjusted odds ratios for several factors potentially influencing vertical transmission for 2periods-1985 through 1995 (January 1, 1985, through December 31,1995) and 1996 through 1999 (January 1, 1996, through December 31, 1999), and between treated and untreated children after 1995.Results: The transmission rate was 15.5% in the 19851995 period and 5.8% in the 1996-1999 period. By 1999, prophylactic interventions had greatly increased. Antiretroviral treatment (ART) usage was 89.9%, (55.1% combination ART) and the elective cesarean delivery rate was 81.3%. In multivariate analysis, only elective cesarean delivery was associated with a lower risk of mother-to-infant transmission before 1995. After 1995, nonbreastfeeding and receipt of ART were protective whereas elective cesarean delivery was not significantly protective in multivariate analysis. Transmission risk was reduced by 76% with an incomplete zidovudine regimen, 88% with a complete regimen, and 93% when the mother received combination ART. In the 1996-1999 period, the transmission rate for nonbreastfeeding mother-infant pairs was 8.6% with elective cesarean delivery, 4.4% with any ART, and 2.4% with these interventions combined.Conclusion: Prophylactic interventions, and in particular ART, reduced perinatal HIV-1 transmission at a population level in Italy

Five-year follow-up of children with perinatal HIV-1 infection receiving early highly active antiretroviral therapy

Background: Early highly active antiretroviral therapy ( HAART), started within the first months of age, has been proven to be the optimal strategy to prevent immunological and clinical deterioration in perinatally HIV-infected children. Nevertheless, data about long-term follow-up of early treated children are lacking. Methods: We report data from 40 perinatally HIV-infected-children receiving early HAART, with a median follow-up period of 5.96 years (interquartile range [IQR]: 4.21-7.62). Children were enrolled at birth in the Italian Register for HIV Infection in Children. Comparison with 91 infected children born in the same period, followed-up from birth, and receiving deferred treatment was also provided. Results: Nineteen children (47.5%) were still receiving their first HAART regimen at last follow-up. In the remaining children the first regimen was discontinued, after a median period of 3.77 years (IQR: 1.71-5.71) because of viral failure (8 cases), liver toxicity (1 case), structured therapy interruption (3 cases), or simplification/switch to a PI-sparing regimen (9 cases). Thirty-nine (97.5%) children showed CD4(+) T-lymphocyte values>25%, and undetectable viral load was reached in 31 (77.5%) children at last visit. Early treated children displayed significantly lower viral load than not-early treated children, until 6 years of age, and higher median CD4(+) T-lymphocyte percentages until 4 years of age. Twenty-seven (29.7%) not-early treated vs. 0/40 early treated children were in clinical category C at last follow-up (P < 0.0001). Conclusion: Our findings suggest that clinical, virologic and immunological advantages from early-HAART are long-lasting. Recommendations indicating the long-term management of early treated children are needed

Lower mother-to-child HIV-1 transmission in boys is independent of type of delivery and antiretroviral prophylaxis. The Italian register for HIV intection in children.

The relationship between infant's gender and rate of HIV-1 mother-to-child transmission (MTCT) was evaluated in a prospective cohort of 4151 children (2166 boys and 1985 girls) born to HIV-1-infected mothers enrolled in the Italian Register for HIV Infection in Children. Logistic regression models were performed to estimate crude odds ratios (ORs) and adjusted odds ratios (AORs) and 95% CIs for factors potentially influencing MTCT separately for the period 1985-1995 and the period 1996-2001. To evaluate rates of MTCT by gender in specific subgroups, separate logistic regression models by mode of delivery and antiretroviral prophylaxis were performed. Among children born in 1985-1995, 15.5% boys (95% CI: 13.6-17.7) and 17.9% girls (95% CI: 15.7-20.3) were infected (P = 0.1181). After 1995, a lower proportion of boys (3.1% [95% CI: 2.0-4.4]; AOR: 0.43 [95% CI: 0.26-0.71], P = 0.0008) than girls (AOR: 6.3%, 95% CI: 4.8-8.1) was infected. Lower AORs for boys persisted independently of elective cesarean delivery (AOR: 0.31, 95% CI: 0.14-0.71); other than elective cesarean (AOR: 0.38, 95% CI: 0.19-0.78) and antiretroviral prophylaxis (zidovudine monotherapy (AOR: 0.11, 95% CI: 0.03-0.38); none (AOR: 0.43, 95% CI: 0.21-0.90). No difference was observed when combined therapy in the mother was administered (AOR: 1.14, 95% CI: 0.30-4.32), but results were likely to be biased by the very low rate of infected children in this group. A lower proportion of HIV-1-infected boys in children born after 1995 was found. Factor(s) intrinsic to gender (rather than type of delivery or maternal antiretroviral prophylaxis) may be involved, because the risk of infection in boys was lower independent of interventions. A possible explanation is that, among infected fetuses, more girls survive up to the end of pregnancy and may take advantage of the benefits of preventive strategies