Dot-Elisa for evaluation of hydatid cyst wall, protoscoleces and hydatid cyst fluid antigens in the serodiagnosis of cystic echinococcosis

The aim of the present study is to evaluate cyst wall and protoscolex as an alternate source of antigen in serodiagnosis of cystic echinococcosis (CE). A total of 90 blood samples, 30 each of confirmed CE cases, disease controls and healthy controls were collected. Dot-ELISA using cyst wall, protoscolex and cyst fluid were used to demonstrate anti-hydatid antibodies. The sensitivity of Dot-ELISA using cyst wall, protoscolex and cyst fluid was 96.66%, 86.66% and 93.33% respectively and the specificity of the assay was 70% for Dot-ELISA using cyst fluid, protoscolex and cyst wall antigens. Results of the present study show that cyst wall and protoscolex can also be an useful source of antigen in detection of hydatid antibodies in the serodiagnosis of CE.

Encefalopatia Hipóxico-Isquémica por Asfixia Perinatal: Evolução a Curto e Longo Prazo

Introdução: A encefalopatia hipoxico-isquémica (EHI) é uma causa importante de mortalidade e morbilidade a longo prazo. Estima-se que anualmente ocorram 1 a 8 casos /1000 nascimentos. A asfixia perinatal mantida é a causa mais frequente de EHI. Objectivo: Avaliar a evolução a curto e longo prazo de crianças com EHI devido a asfixia perinatal. Metodologia: Estudo prospectivo histórico. População: Recém-nascidos (RN) consecutivos, nascidos na maternidade do HDE, com idade gestacional ≥ 37 semanas, admitidos na UCIN com os diagnósticos de EHI e asfixia perinatal, sem anomalias congénitas major. Período: 04/04/2001 – 31/12/2010. Variáveis: morbilidade, sobrevivência e sequelas. Covariáveis: peso ao nascer, idade gestacional, índice de Apgar, eventos peri-parto e tipo de parto. Resultados: Dos 19365 nados-vivos (NV), foram incluídos 28 RN (incidência 1,4/1000). Características da amostra: 68 % do sexo masculino; peso médio ao nascer 3180 g, mediana da idade gestacional 39,4 semanas. Índice de Apgar: moda ao 1º, 5º e 10º minuto, respectivamente 2, 5 e 7. Eventos peri-parto mais frequentes: líquido amniótico meconial (46,4%), alterações cardiotocográficas (50%) e circular cervical apertada (10,7%). Em 67,8% dos casos, foi realizada cesariana de emergência. Todos os RN necessitaram de manobras de reanimação. Desenvolveram EHI: ligeira 12 (42,9%), moderada 7 (25%) e grave 9 (32,1%). Ocorreram convulsões em 66% dos RN e disfunção multiorgânica em 12 RN (42,9%). Dos exames de imagem, a ecografia transfontanelar (ECOTF) foi realizada em 89,3% dos casos, o electroencefalograma (EEG) em 67,9% e a RMN em 32,1%. A ECOTF revelou alterações sugestivas de EHI em todos os casos; o EEG evidenciou anomalia da actividade eléctrica em 73,7% e a RMN revelou achados compatíveis com EHI em 100%. Houve 3 óbitos. Dos sobreviventes, 75% foram seguidos em consulta hospitalar, apresentando como sequelas graves paralisia cerebral (22,2%), atraso global do desenvolvimento (16,6%), epilepsia (11,1%), surdez neurossensorial (5,6%) e hemiparésia (5,6%). Conclusões: A incidência de EHI por asfixia foi 1,4/1000 NV. Verificou-se EHI ligeira em 42,8% dos casos. A mortalidade foi de 14,3%. Nos 18 casos seguidos, 38,9% não apresentavam sequelas ou tinham sequelas ligeiras. Apenas um caso foi submetido a hipotermia, sendo esta uma terapêutica promissora.

Perinatal Bacterial Infection: Screenning of Vertical Transmitted Infections

Perinatal bacterial infection may be caused by any microorganism colonizing the vaginal tract. Neonatologists and paediatricians are especially concerned about group B Stretpococcus (GBS). However, Enterobactereacea, mainly E.coli and Proteus, are also responsible for infection. GBS screening may be accomplished in over 90% of pregnant women. In our maternity in 2007-2008, 85% of the mothers had been screened. Screening and prophylaxis were responsible for a decreasing incidence of neonatal infection - from 0.6/1000 to 0.15/1000 live births in Portugal, from 2002 to 2007. However there are some difficulties related to screening. In the second Portuguese study 16/57 NB with early-onset infection (28%) were born to “negative” mothers. Several factors illustrate how difficult is to draw national screening policies: a wide range of carrier’s state rate throughout a country - in Portugal from 12% to 30%. The success of any screening policy may also be affected by additional technical and organizational problems. In countries where home delivery is a tradition or a trend intrapartum GBS prophylaxis requires a very well organized assistance.. Moreover factors usually accepted as protective are not so effective. In the Portuguese study 24/57 infected newborns (42%) were delivery by caesarean section. Another subject deals with the workload in the postnatal ward generated by deficient compliance to the guidelines a problem not confirm by a study of our group. Decreasing the importance of GBS, highlight the importance of E. coli in perinatal infection. From the 16 340 registrations of the National Registry 1676 were newborns with mother-related infection. Applying the same reasoning to E.coli as to GBS and Listeria monocytogenes – that is considering all of them are of maternal origin - 6.7% of these infections were due to E. coli, 4.6% to SGB and 0.5% to Listeria monocytogenes. In conclusion screening and prophylaxis may be not the best way to prevent all GBS neonatal infections but by now it is the only available procedure. The other bacteria continue to demand a high suspicion level and immediate intervention.

Perinatal Death: Epidemiology and Etiology

Perinatal mortality rate is an important mark to evaluate women and perinatal health care. It is of utmost importance to know causes and the evolution of its two components aiming to improve health care in different fields – sanitary conditions, diagnosis and treatment of infectious disease, immunisations, diagnosing and caring for medical diseases induced by pregnancy or directly related to it, providing skilled birth attendance, preventing birth asphyxia, preventing preterm birth complications and infections. In high-income countries the epidemiology varies mainly with social and economic conditions; in low-income countries, paired with poverty, undernutrition, superstition, lack of medical care, deficient basic sanitary conditions are also found. Also, in rich countries, responsible for 1% of deaths, data are published and improvements evaluated, while in low-income countries responsible for 99% of deaths numbers and causes are unknown, making difficult to implement cost effective interventions, a reason why “stillbirth rates in low-income countries are now where they were in high-income countries 50 to 100 years ago”. Knowledge on causes of death are very important as often what is needed are “simple” measures as improvement of sanitary conditions and immunisation programmes rather than high technologies. About four million babies dye each year in the first 28 days of life and another 3 million dye before birth in the third-trimester, with 98% occurring in low-income and middle income countries and more than 1 million occurring during labour and delivery. Classically stillbirths are the major component of perinatal mortality rate. Causes of death are even more difficult to know. In low-income countries a great proportion of women give birth at home. Worldwide the main causes of stillbirth are asphyxia due to obstructed labour, eclampsia, abruption placenta and umbilical cord complications - making valid the assumption that skilled birth attendance would decrease stillbirth; and infection - chorioamnioitis, syphilis and malaria. In high-income countries placental pathology and infection, congenital anomalies, complications of preterm birth and post term delivery, are the most common. If in low-income countries famine and lack of provisions and health care are common, in high-income countries, advanced maternal age and diabetes, obesity, hypertension, smoking, are frequent findings.

Induced Labor– Is There an Increased Risk of Perinatal Infection?

Objectives: To assess induced labor-associated perinatal infection risk at Hospital D.Estefânia from January to June of 2010 at Hospital de D. Estefânia’s delivery rooms, reviewing the indications for inducing labor as well as the techniques used. Material and Methods: Performing an historical prospective study searching the clinical processes as well as the mother and newborn’s computer database from January to June of 2010. An exposed and an unexposed group were created; the first group comprises pregnant women and their newborns whose labor was induced. The unexposed group is constituted by newborns and pregnant women whose labor was spontaneous. Labor induction was performed using intra-vaginal prostaglandins in women who didn’t start it spontaneously; perinatal infection was defined either clinically or using blood tests. The gestational age was ≥ 37 weeks for both groups. 19 variables were studied for both groups. Results: A total of 190 mother-newborn pairs were included: 55 in the exposed group and 135 in the unexposed group. 3 cases of perinatal infection were reported, two in the exposed group and one in the unexposed group. Preliminary data resulted in a perinatal infection rate of 3.6% in the exposed group and 0.7% in the unexposed group; preliminary data suggest that the risk of perinatal infection may be increased in up to 5-fold when labor is inducted. Conclusions: A larger series of patients and a multivariable analysis using logistic regression are both necessary in order to perform a more thorough assessment of labor induction’s role in perinatal infection risk. One must also try to distinguish labor inducing- and clinical practicesrelated factors.

Serological, clinical and epidemiological evaluation of toxocariasis in urban areas of south Brazil

Toxocariasis is a worldwide public-health problem that poses major risks to children who may accidentally ingest embryonated eggs of Toxocara. The objectives of this study were to investigate the occurrence of anti-Toxocara spp. antibodies in children and adolescents and the variables that may be involved, as well as environmental contamination by Toxocara spp. eggs, in urban recreation areas of north central mesoregion, Paraná State, Brazil. From June 2005 to March 2007. a total of 376 blood samples were collected by the Public Health Service from children and adolescents one to 12 years old, of both genders. Samples were analyzed by the indirect ELISA method for detection of anti-Toxocara antibodies. Serum samples were previously absorbed with Ascaris suum antigens, and considered positive with a reagent reactivity index >1. Soil samples from all of the public squares and schools located in the four evaluated municipalities that had sand surfaces (n = 19) or lawns (n = 15) were analyzed. Of the 376 serum samples, 194 (51.6%) were positive. The seroprevalence rate was substantially higher among children aging one to five years (p = 0.001) and six to eight years (p = 0.022). The clinical signs and symptoms investigated did not show a statistical difference between seropositive and seronegative individuals (p > 0.05). In 76.5% of the investigated recreation places, eggs of Toxocara were detected in at least one of the five collected samples. Recreation areas from public schools were 2.8 times more contaminated than from public squares. It is important to institute educational programs to inform families and educators, as well as to improve sanitary control of animals and cleaning of the areas intended for recreation in order to prevent toxocariasis.

Evaluation of Helicobacter pylory colonization by serologic test (IgG) and dyspepsia in volunteers from the countryside of Monte Negro, in the Brazilian western Amazon region

The present study intended to analyze the seroprevalence of Helicobacter pylori, IgG, and its relation to dyspepsia in a population from the western Amazon region. During the "Projeto Bandeira Científica", a University of São Paulo Medical School program, in Monte Negro's rural areas, state of Rondônia, 266 blood samples were collected from volunteers. The material was tested for IgG antibodies anti-Helicobacter pylori by ELISA method and the participants were also interviewed on dyspepsia, hygiene and social aspects. Participants aged between five and 81 years old (34 years on average), 149 (56%) were female and 117 (44%) male. We found 210 (78.9%) positive, 50 (18.8%) negative and six (2.3%) undetermined samples. Dyspeptic complaints were found in 226 cases (85.2%). There was no statistical association between dyspepsia and positive serology for H. pylori. We concluded that the seroprevalence in all age categories is similar to results found in other studies conducted in developing countries, including those from Brazil. On the other hand, the seroprevalence found in Monte Negro was higher than that reported in developed countries. As expected, there was a progressive increase in the positivity for H. pylori in older age groups.

Regionalización del Cuidado Perinatal en Portugal - Resultados

Lo Servicio Nacional de Salud en Portugal fue criado en 1979, universal e gratis. Las primeras UCIN surgieran en 1980, en 1985 fue criada la Sociedad Portuguesa de Neonatología, en 1987 lo sistema de transporte neonatal, en mismo año fue nominada una Comisión de Peritos en Perinatología. En 1989 fue nominado el Comité Nacional de la Mujer y del niño y empezó la Reforma de los cuidados de salud perinatal. Era un programa de 9 años en etapas de 3 años que incluía el ccierre de Hospitales con menos de 1500 partos/año, categorización de los hospitales en niveles de cuidados e la creación de Unidades Coordinadoras entre Centros de Salud y Hospitales. Las UCIN y Intermedios neonatales fueran equipados y definido el número necesario de obstetras, pediatras y enfermeras e fue hecha formación en Cuidados Intensivos Neonatales. Los Centros de Salud no tienen partos e controlan el embarazo normal; los hospitales Nivel I no tienen partos; los de Nivel II tienen partos normales y de bajo riesgo, por lo menos 1500/año, obstetras, Unidad de Cuidados Intermedios, Pediatras con formación en neonatología, ventilación por períodos cortos. Los Hospitales de Nivel III tienen partos de bajo y alto riesgo, obstetras y neonatólogos, UCIN, formación en obstetricia y neonatología e investigación. Las UCIN tienen ventilación de longa duración, nutrición parenteral, cuidados de recién nacidos con menos de 1500g, condiciones quirúrgicas, son centros de enseñanza e investigación. Deben tener neonatólogos y pediatras con competencia en neonatología 24h por día, 1.5 camas /1000 partos, 1 enfermera para 2 logares de intensivos – 2,5 enfermeras por cada cama de intensivos - deben estar localizadas en maternidades con >3000 partos. Hay también reglas para translado para nível III pré-natal y pós-natal. Lo impacto fue que la tasa de partos hospitalares aumentó hasta 99% la mortalidad fetal e fetal tardia disminuiu hasta 3,7 e 2,5/1000 NV+MN, la mortalidad perinatal con más de 28 semanas para 4,6/1000 NV+ MN, la neonatal para 2,4/1000NV e la infantil para 3,6. Las mejores condiciones socioeconómicas y nivel de educación pueden justificar parte de estos resultados pero la organización de los cuidado perinatales fue ciertamente una grande razón de mejoría.

Why and How Was Regionalization of Perinatal Care Accomplished in Portugal?

Portuguese health care system was created in 1979. It is universal and for free. Expenses are supported by the State through taxes. The modern perinatal care system started by the end of 1970. The first neonatal intensive care units were created in 1980, the Portuguese Neonatal Society in 1985 and the National Neonatal Transport System in 1987. Until the seventies of twentieth century and even during eighties there were more than 200 hospitals with deliveries, a great part without obstetrician or paediatrician, a great percentage of pregnancies had no prenatal care, there were few neonatal intensive care units and perinatal mortality rate was one of the highest in the European countries. In 1987 an Experts Committee was nominated by the Health Ministry aiming to collect and analyse data on perinatal care and to suggest improvements. The Report resulting from this work is the main document on which is based the reform. The reform was a 9 years program in 3 years stages aiming to close hospitals with less than 1500 deliveries/year, to reclassify hospitals, to create Coordinating Units between health centres and hospitals, to equip neonatal intensive and intermediate care units, to define needs of obstetricians, paediatricians and nurses for each centre and to promote specialised training in neonatology for paediatricians and nurses. Levels of perinatal care were defined as well as localization of each level of hospital according to the number of deliveries in one geographic area, geographic difficulties and existing routes and connections. Steps for opening and closure of different levels of hospitals were very well programmed. The organization, capacities, number of obstetricians, neonatologists and nurses as well as equipment for each level of care was defined. Rules for pregnant women and newborns transfer from level II to level III hospitals were also well described. A specific training is neonatology was created starting in 1990. This organization resulted in an impressive decrease in mortality rates at all levels and still it is the policy we have today.

Evaluation of enterovirus 71 immune status in São Paulo state, Brazil

Antibodies to Enterovirus 71 (EV71) were evaluated in São Paulo State during 1999-2005. The titer of neutralizing antibodies against EV71 was determined by microneutralization assay, and a titer of > 1:8 was defined as indicative of protected immunity. Neutralizing antibodies to EV71 were observed in 12.4% (55/442) of sera samples, a low protective rate, suggesting that EV71 infection is uncommon in this region, but that there is a relatively high susceptibility to EV71 related diseases, which is worrying considering the recent Asian outbreaks. Also, a significant location-specific difference in seropositivity was observed. Neutralizing antibodies to EV71 were observed in 8.7% (21/241) of São Paulo metropolitan area sera samples, and 16.9% (34/201) of the sera samples from other municipalities. A high number of Brazilian residents live in country and coastal areas without adequate access to piped water or sanitation. This situation may contribute to the EV71 dissemination in these zones. The analysis of environmental samples could possibly make a valuable contribution to studies on the epidemiology of EV71.

Evaluation of glycoprotein B genotypes and load of CMV infecting blood leukocytes on prognosis of AIDS patients

BACKGROUND: Cytomegalovirus (CMV) remains an important pathogen to immunocompromised patients even in the era of HAART. The present study aimed at evaluating the influence of CMV viral load and its gB genotypes on AIDS patients' outcome. METHODS: Blood samples of 101 AIDS patients were collected and tested for HIV load, CD4 - cell count and opportunistic pathogens, including CMV. Semi-nested PCRs were run to detect CMV genome and in the positive samples, gB genotyping and CMV load were established using enzymatic restriction and real time PCR, respectively. All patients were clinically followed for four years. RESULTS: In thirty patients (31%) CMV was detected and all fatal cases (n = 5) occurred in this group of patients (p = 0.007), but only two patients had CMV disease (1.9%). However, viral load was not statistically associated with any analyzed parameter. The most frequently observed CMV genotype was gB2 (45.16%) followed by gB3 (35.48%). gB2 genotype was more frequently found in patients with CD4-cell counts under 200 cells/mm³ (p = 0.0017), and almost all fatal cases (80%) had gB2 genotype. CONCLUSIONS: Our study suggests that CMV and its polymorphisms in biologically relevant genes, such as the gB encoding ORF, may still influence the prognosis and outcome of AIDS patients. The gB2 genotype was associated to patient's bad outcome.

Evaluation of dengue NS1 antigen detection for diagnosis in public health laboratories, São Paulo State, 2009

The present work evaluated the diagnostic accuracy of detection of Dengue NS1 antigen employing two NS1 assays, an immunochromatographic assay and ELISA, in the diagnostic routine of Public Health laboratories. The results obtained with NS1 assay were compared with virus isolation and, in a subpopulation of cases, they were compared with the IgM-ELISA results obtained with convalescent samples. A total of 2,321 sera samples were analyzed by one of two NS1 techniques from March to October 2009. The samples were divided into five groups: groups I, II and III included samples tested by NS1 and virus isolation, and groups IV and V included patients with a first sample tested by NS1 and a second sample tested by IgM-ELISA. Sensitivity, specificity, positive and negative predictive values, Kappa Index and Kappa Concordance were calculated. The results showed that NS1 testing in groups I, II and III had high sensitivity (98.0%, 99.5% and 99.3%), and predictive values and Kappa index between 0.9 - 1.0. Groups IV and V only had Kappa Concordance calculated, since the samples were analyzed according to the presence of NS1 antigen or IgM antibody. Concordance of 92.1% was observed when comparing the results of NS1-negative samples with IgM-ELISA. Based on the findings, it is possible to suggest that the tests for NS1 detection may be important tools for monitoring the introduction and spread of Dengue serotypes.

Evaluation of GBV-C / HVG viremia in HIV-infected women

The present study aimed at standardizing a real-time quantitative polymerase chain reaction assay to evaluate the presence of GBV-C/HGV RNA. A "TaqMan" assay using primers and probe derived from the 5¢ NCR region was developed and validated. Two hundred and fifty-three plasma samples from HIV-infected women were tested for GBV-C viremia and antibody against the envelope protein 2. GBV-C RNA was detected in 22.5% of the patients whereas the antibody was identified in 25.3% of the cohort. Detection of viral RNA and of antibodies was mutually exclusive. Viral loads showed a mean of 1,777 arbitrary units / mL, being 1.1 and 13,625 arbitrary units / mL respectively the lowest and highest values measured. We conclude that the real-time quantitative polymerase chain reaction method developed is appropriate for the investigation of GBV-C RNA since it was shown to be highly specific and sensitive, as well as requiring few steps, preventing contamination and providing additional information as to the relative viremia of carriers, a parameter that must be included in studies evaluating the co-factors influencing the clinical outcome of HIV/AIDS.

Perinatal Outcome and Change in Body Mass Index in Mothers of Dichorionic Twins: a Longitudinal Cohort Study

We used a prospective cohort to analyze the effect of change in BMI rather than change in weight, in mothers carrying dichorionic twins from a population that did not receive any dietary intervention. A total of 269 mothers (150 nulliparas and 119 multiparas) were evaluated. The average change (%) from the pre-gravid BMI was 7.2+/-6.1, 17.4+/-8.2, and 28.7+/-10.8, at 12-14, 22-25, and 30-34 weeks, respectively, without difference between nulliparas and multiparas. The comparison between maternities below or above the average change from the pregravid BMI failed to demonstrate an advantage (in terms of total twin birthweight and gestational age) of an above average change from the pregravid BMI, even when the lower versus upper quartiles were compared. Our observations reached different conclusions regarding the recommended universal dietary intervention in twin gestations. A cautious approach is advocated towards seemingly harmless excess weight gain, as normal weight women may turn overweight, or even obese, by the end of pregnancy, and be exposed to the untoward effects of obesity on future health and body image.