Maria Célia Cervi

Congenital and perinatal cytomegalovirus infection in infants born to mothers infected with human immunodeficiency virus

OBJECTIVES To determine the rates of congenital and perinatal cytomegalovirus (CMV) infection among infants born to mothers infected with HIV compared with infants born to mothers not infected with HIV from a CMV-immune, low-income population. STUDY DESIGN A total of 325 newborns from CMV-seropositive mothers were enrolled and evaluated for congenital CMV infection (150 infants from HIV+ mothers and 175 infants from HIV- mothers. A total of 101 infants from HIV+ mothers and 33 infants from HIV- mothers were evaluated for perinatal CMV infection. The virus was isolated from urine by culture in human fibroblasts and was detected by polymerase chain reaction at birth and at 15 days and 12 weeks of age. RESULTS Only 13 of 150 HIV+ mothers (8.7%) had an AIDS-defining condition, and none had a late-stage HIV infection. Congenital CMV infection was detected in 4 of 150 (2.7%) infants from HIV+ mothers and in 5 of 175 (2.9%) infants from HIV- mothers (p = 1.00). Perinatal CMV infection was diagnosed in 8 of 101 (7.9%) infants from HIV+ mothers and in 13 of 33 (39.4%) infants from HIV- mothers (p < 0.00001). Most infants (93.9%) from HIV- mothers and only 5.9% of infants from HIV+ mothers were breastfed. CONCLUSIONS CMV coinfection in mothers without advanced HIV disease from a CMV-immune population does not enhance the likelihood of congenital CMV infection. Perinatal CMV transmission from HIV-infected mothers may be decreased by avoiding breastfeeding. Further studies on mothers with late-stage HIV infection are needed.

ADA-deficient SCID is associated with a specific microenvironment and bone phenotype characterized by RANKL/OPG imbalance and osteoblast insufficiency.

Adenosine deaminase (ADA) deficiency is a disorder of the purine metabolism leading to combined immunodeficiency and systemic alterations, including skeletal abnormalities. We report that ADA deficiency in mice causes a specific bone phenotype characterized by alterations of structural properties and impaired mechanical competence. These alterations are the combined result of an imbalanced receptor activator of nuclear factor-kappaB ligand (RANKL)/osteoprotegerin axis, causing decreased osteoclastogenesis and an intrinsic defect of osteoblast function with subsequent low bone formation. In vitro, osteoblasts lacking ADA displayed an altered transcriptional profile and growth reduction. Furthermore, the bone marrow microenvironment of ADA-deficient mice showed a reduced capacity to support in vitro and in vivo hematopoiesis. Treatment of ADA-deficient neonatal mice with enzyme replacement therapy, bone marrow transplantation, or gene therapy resulted in full recovery of the altered bone parameters. Remarkably, untreated ADA-severe combined immunodeficiency patients showed a similar imbalance in RANKL/osteoprotegerin levels alongside severe growth retardation. Gene therapy with ADA-transduced hematopoietic stem cells increased serum RANKL levels and childrens growth. Our results indicate that the ADA metabolism represents a crucial modulatory factor of bone cell activities and remodeling.

Detection of Human Bocavirus mRNA in Respiratory Secretions Correlates with High Viral Load and Concurrent Diarrhea

Human bocavirus (HBoV) is a parvovirus recently identified in association with acute respiratory infections (ARI). Despite its worldwide occurrence, little is known on the pathogenesis of HBoV infections. In addition, few systematic studies of HBoV in ARI have been conducted in Latin America. Therefore, in order to test whether active viral replication of human bocavirus is associated with respiratory diseases and to understand the clinical impact of this virus in patients with these diseases, we performed a 3-year retrospective hospital-based study of HBoV in outpatients and inpatients with symptoms of Acute Respiratory Infections (ARI) in Brazil. Nasopharyngeal aspirates (NPAs) from 1015 patients with respiratory symptoms were tested for HBoV DNA by PCR. All samples positive for HBoV were tested by PCR for all other respiratory viruses, had HBoV viral loads determined by quantitative real time PCR and, when possible, were tested by RT-PCR for HBoV VP1 mRNA, as evidence of active viral replication. HBoV was detected in 4.8% of patients, with annual rates of 10.0%, 3.0% and 3.0% in 2005, 2006 and 2007, respectively. The range of respiratory symptoms was similar between HBoV-positive and HBoV-negative ARI patients. However, a higher rate of diarrhea was observed in HBoV-positive patients. High HBoV viral loads (>108 copies/mL) and diarrhea were significantly more frequent in patients with exclusive infection by HBoV and in patients with detection of HBoV VP1 mRNA than in patients with viral co-infection, detected in 72.9% of patients with HBoV. In summary, our data demonstrated that active HBoV replication was detected in a small percentage of patients with ARI and was correlated with concurrent diarrhea and lack of other viral co-infections.

Nasopharyngeal colonization with Streptococcus pneumoniae in children infected with human immunodeficiency virus

OBJECTIVES To determine the prevalence of pneumococcus colonization among HIV-infected outpatients aged 0 to 18 years. To determine the resistance to penicillin of the microorganisms observed, to identify their serotypes, and to determine whether there are associations between known risk factors and colonization in this group. MATERIAL AND METHOD This was an observational and cross-sectional study in which nasopharynx swabs were collected from 112 children on the occasion of their monthly appointments and a questionnaire applied to the mothers. The material collected was processed at the microbiology laboratory of the hospital in accordance with National Committee for Clinical Laboratory Standards (NCCLS) regulations and serotyping was performed at the Centers for Diseases Control and Prevention (CDC). Data were analyzed statistically using the chi-square test and with univariate and multivariate analysis with multiple logistic regression. RESULTS The prevalence rate of nasopharyngeal colonization by pneumococci was 28.6%, with a 15.6% rate of resistance to penicillin (6.2% intermediate resistance and 9.4% full resistance). The serotypes identified were 6A, 6B, 7C, 9V, 11A, 13, 14, 15A, 16F, 18C, 19B, 19F, 23B, 23F, and 34. In this population there were no associations between pneumococcal colonization and the risk factors studied. CONCLUSIONS The prevalence of pneumococcal colonization among HIV-infected children at our service was not higher than prevalence rates observed in healthy children and reported in the literature.

Rubella Vaccination of Unknowingly Pregnant Women: The São Paulo Experience, 2001

BACKGROUND Rubella vaccination is contraindicated during pregnancy. During mass immunization of women of childbearing age against rubella, women unknowingly pregnant may be vaccinated. To evaluate the effects of rubella vaccination during pregnancy, the Brazilian state of São Paulo conducted a follow-up study of pregnant women vaccinated during a rubella campaign in 2001. METHODS Women vaccinated during pregnancy were reported to a national surveillance system. In the state of São Paulo, follow-up of vaccinated women included household interviews. Serum samples from vaccinated women were tested for antirubella antibodies to classify susceptibility to rubella infection. Children born to susceptible mothers were tested for evidence of congenital rubella infection and evaluated for signs of congenital rubella syndrome. RESULTS The São Paulo State Health Department received 6473 notifications of women vaccinated during pregnancy. Serology performed for 5580 women identified 811 (15%) that were previously susceptible. Incidence of spontaneous abortion or stillbirth among previously susceptible vaccinated women was similar to women with prior immunity. Twenty-seven (4.7%) of 580 newborns tested had evidence of congenital rubella infection; none had congenital rubella syndrome. CONCLUSIONS Mass rubella vaccination of women of childbearing age was not associated with adverse birth outcomes or congenital rubella syndrome among children born to women vaccinated during pregnancy.

Pneumococcal nasopharyngeal carriage among infants born to human immunodeficiency virus-infected mothers immunized with pneumococcal polysaccharide vaccine during gestation.

Background: We have previously shown that 23-valent pneumococcal polysaccharide vaccine (PPV) is immunogenic in human immunodeficiency virus (HIV)-infected mothers and provides vaccine-induced antibodies to the infant. We compared the nasopharyngeal pneumococcal colonization (NPC) rates in <6-month-old infants born to HIV-infected mothers, according to immunization with PPV during pregnancy. Methods: NPC was evaluated in 45 term infants born to vaccinated women (PPV+) and in 60 infants in a control group (PPV−), at 2 months (±30 days), 4 months (±30 days), and 6 months (±30 days) of age. Results: A total of 82 infants completed the study (at least 2 of 3 evaluations), 35 (77%) in the PPV+ and 47 (78.3%) in the PPV− groups, respectively. Infant gender, HIV infection status, number of adults, children, and smokers in the household, day-care attendance, occurrence of respiratory signs, and cotrimoxazole use were similar in both groups. NPC rates increased equally with age in both groups (2 months = 26.7% vs. 25.6%; 4 months = 34.5% vs. 38.6%; 6 months = 38.7% vs. 56.3%, in PPV+ and PPV−, respectively). After controlling for potential confounders, we found no association between maternal vaccination and infant pneumococcal carriage (adjusted odds ratio = 0.70; 95% confidence interval: 0.23, 2.21) Conclusions: Vaccination of HIV-infected mothers with PPV did not protect infants younger than 6 months of age from nasopharyngeal pneumococcal carriage.

Vertical transmission of Cryptococcus neoformans from a mother coinfected with human immunodeficiency virus: case report

Disseminated infection with Cryptococcus neoformans was observed in a newborn infant who presented fever and respiratory symptoms since the 52 nd day of life. The mother was infected by human immunodeficiency virus and presented pulmonary and meningeal cryptococcal infection. This is a rare case of cryptococcal infection with probable maternal-fetal transmission.

Colonização nasofaríngea por Streptococcus pneumoniae em crianças com infecção pelo vírus da imunodeficiência humana

OBJECTIVE: To determine the prevalence of pneumococcus colonization among HIV-infected outpatients aged 0 to 18 years. To determine the resistance to penicillin of the microorganisms observed, to identify their serotypes, and to determine whether there are associations between known risk factors and colonization in this group. MATERIAL AND METHOD: This was an observational and cross-sectional study in which nasopharynx swabs were collected from 112 children on the occasion of their monthly appointments and a questionnaire applied to the mothers. The material collected was processed at the microbiology laboratory of the hospital in accordance with National Committee for Clinical Laboratory Standards (NCCLS) regulations and serotyping was performed at the Centers for Diseases Control and Prevention (CDC). Data were analyzed statistically using the chi-square test and with univariate and multivariate analysis with multiple logistic regression. RESULTS: The prevalence rate of nasopharyngeal colonization by pneumococci was 28.6%, with a 15.6% rate of resistance to penicillin (6.2% intermediate resistance and 9.4% full resistance). The serotypes identified were 6A, 6B, 7C, 9V, 11A, 13, 14, 15A, 16F, 18C, 19B, 19F, 23B, 23F, and 34. In this population there were no associations between pneumococcal colonization and the risk factors studied. CONCLUSIONS: The prevalence of pneumococcal colonization among HIV-infected children at our service was not higher than prevalence rates observed in healthy children and reported in the literature.

Concurrent detection of other respiratory viruses in children shedding viable human respiratory syncytial virus.

Human respiratory syncytial virus (HRSV) is an important cause of respiratory disease. The majority of studies addressing the importance of virus co‐infections to the HRSV‐disease have been based on the detection of HRSV by RT‐PCR, which may not distinguish current replication from prolonged shedding of remnant RNA from previous HRSV infections. To assess whether co‐detections of other common respiratory viruses are associated with increased severity of HRSV illnesses from patients who were shedding viable‐HRSV, nasopharyngeal aspirates from children younger than 5 years who sought medical care for respiratory infections in Ribeirão Preto (Brazil) were tested for HRSV by immunofluorescence, RT‐PCR and virus isolation in cell culture. All samples with viable‐HRSV were tested further by PCR for other respiratory viruses. HRSV‐disease severity was assessed by a clinical score scale. A total of 266 samples from 247 children were collected and 111 (42%) were HRSV‐positive. HRSV was isolated from 70 (63%), and 52 (74%) of them were positive for at least one additional virus. HRSV‐positive diseases were more severe than HRSV‐negative ones, but there was no difference in disease severity between patients with viable‐HRSV and those HRSV‐positives by RT‐PCR. Co‐detection of other viruses did not correlate with increased disease severity. HRSV isolation in cell culture does not seem to be superior to RT‐PCR to distinguish infections associated with HRSV replication in studies of clinical impact of HRSV. A high rate of co‐detection of other respiratory viruses was found in samples with viable‐HRSV, but this was not associated with more severe HRSV infection. J Med. Virol. 85:1852–1859, 2013.

Impact of hypoxia on IGF-I, IGF-II, IGFBP-3, ALS and IGFBP-1 regulation and on IGF1R gene expression in children

UNLABELLED Hypoxia is one of many factors involved in the regulation of the IGF system. However, no information is available regarding the regulation of the IGF system by acute hypoxia in humans. OBJECTIVE The aim of this study was to evaluate the effect of acute hypoxia on the IGF system of children. DESIGN Twenty-seven previously health children (14 boys and 13 girls) aged 15 days to 9.5 years were studied in two different situations: during a hypoxemic state (HS) due to acute respiratory distress and after full recovery to a normoxemic state (NS). In these two situations oxygen saturation was assessed with a pulse-oximeter and blood samples were collected for serum IGF-I, IGF-II, IGFBP-1, IGFBP-3, ALS and insulin determination by ELISA; fluoroimmunometric assay determination for GH and also for IGF1R gene expression analysis in peripheral lymphocytes by quantitative real-time PCR. Data were paired and analyzed by the Wilcoxon non-parametric test. RESULTS Oxygen saturation was significantly lower during HS than in NS (P<0.0001). IGF-I and IGF-II levels were lower during HS than in NS (P<0.0001 and P=0.0004, respectively). IGFBP-3 levels were also lower in HS than in NS (P=0.0002) while ALS and basal GH levels were higher during HS (P=0.0015 and P=0.014, respectively). Moreover, IGFBP-1 levels were higher during HS than in NS (P=0.004). No difference was found regarding insulin levels. The expression of IGF1R mRNA as 2(-ΔΔCT) was higher during HS than in NS (P=0.03). CONCLUSION The above results confirm a role of hypoxia in the regulation of the IGF system also in humans. This effect could be direct on the liver and/or mediated by GH and it is not restricted to the hepatocytes but involves other cell lines. During acute hypoxia a combination of alterations usually associated with reduced IGF action was observed. The higher expression of IGF1R mRNA may reflect an up-regulation of the transcriptional process.