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We evaluated the efficacy of noninvasive fetal Rhesus D (RHD) genotyping from maternal plasma in a highly admixed population. Fifty-five blood samples from RhD-negative pregnant women from Brazil were processed for extraction of cell-free plasma DNA. Real-time PCR was performed to amplify segments of exons 5 and 7 from the RHD gene, as well as for detection of the SRY gene to confirm the presence of fetal DNA. Fetal genotyping results were compared with the RhD phenotype determined from newborn cord blood samples obtained at birth. Thirty-two samples were RHD-positive, 18 were RHD-negative and 5 were inconclusive due to amplification of only one RHD exon. In 43 samples, the fetal RHD genotype was compared to the neonatal RhD phenotype, and only one result was discordant, due to false-negative serology. There was one false SRY genotyping negative result. We conclude that noninvasive fetal RHD genotyping from maternal blood provides accurate results and suggests its viability as a clinical tool for the management of RhD-negative pregnant women in an admixed population.

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Cryopreservation of mesenchymal stem cells from amniotic fluid is of clinical importance, as these cells can be harvested during the prenatal period and stored for use in treatments. We examined the behavior of mesenchymal stem cells from human amniotic fluid in culture that had been subjected to cryopreservation. We assessed chromosomal stability through karyotype analysis, determined whether multipotent capacity (differentiation into adipogenic, chondrogenic, and osteogenic cells) is maintained, and analyzed SOX2 and NANOG expression after thawing. Five amniotic fluid samples were cryopreserved for 150 days. No chromosomal aberrations were observed. The expression levels of NANOG and SOX2 also were quite similar before and after cryopreservation. Capacity for differentiation into adipogenic, chondrogenic, and osteogenic tissues also remained the same. We conclude that cryopreservation of amniotic fluid does not alter karyotype, NANOG/SOX2 gene expression, or multipotent capacity of stem cells that have been collected from amniotic fluid during pregnancy.

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Malaria in Brazil is virtually restricted to the Amazon Region, where it has a heterogeneous geographic distribution. We reviewed secondary data in order to describe the regional and temporal distribution of 8018 malaria cases seen between 2003 and 2007 in Santa Isabel do Rio Negro, a municipality in the northwest Brazilian Amazon. A significant rise in malaria incidence, mainly in the Yanomami Indian reservation, was observed during this time. Anopheline breeding sites were also mapped and entomological data were obtained through the capture of larval and adult mosquitoes. Thirty-three potential breeding sites were identified in the urban and periurban areas, 28 of which were positive for anopheline larvae. Anopheles darlingi specimens were captured in both intra- and peridomicile locations in the urban areas. Demographic data were also assessed via a sectional survey, revealing that the majority of dwellings were vulnerable to mosquitoes. This study suggests that urban and periurban areas of this municipality are highly susceptible to epidemic malaria, which is endemic in the Yanomami Indian reservation near the city. In addition, transmission can be perpetuated autochthonously in the urban area, drawing attention to the continuous need for preventative measures such as controlling adult and aquatic stages of mosquitoes and improving housing.

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OBJECTIVE: To verify the correlation between fetal splenic artery Doppler velocimetry and fetal hemoglobin (Hb) levels in Rh alloimmunization. METHODS: Splenic artery Doppler peak systolic velocity (PSV) and pulsatility index (PI) were obtained before cordocentesis in rhesus-alloimmunized fetuses. Doppler was performed before 80 cordocentesis in 36 patients between 20 and 35 weeks of gestation. Mild, moderate and severe anemia were defined as a Hb deficit of >or=2, >or=5 and >or=7 g/dl respectively. RESULTS: Anemia was noted in 64% of the fetuses and moderate and severe anemia in 18 and 21%. Splenic artery PSV was higher in groups with moderate (p = 0.001) and severe (p < 0.000) anemia but not in the group with mild anemia (p = 0.189) when compared to non-anemic fetuses. Splenic artery PI was higher only in the severely anemic group (p = 0.001). CONCLUSIONS: The splenic artery PI and PSV are higher in fetuses with severe anemia.

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OBJECTIVE: To test a new noninvasive ultrasound method for diagnosing fetal anemia in red blood cell isoimmunized pregnancies. METHODS: A diagnostic accuracy study was carried out to determine the cutoff point of an ultrasound measurement, the cardiofemoral index (CFI), calculated using the biventricular outer dimension (BVOD) and femur length to diagnosis severe anemia. The CFI measurement was performed before each of the 336 cordocenteses on 131 fetuses. Diagnosis test analysis and receiver-operating characteristics (ROC) curves were used and the area under the curve (AUC) was calculated to compare the overall accuracy of the CFI for anemia diagnosis, between fetuses with or without previous intrauterine transfusions (IUT). RESULTS: At first cordocentesis (n=131) the AUC was 0.75 (95% CI, 0.66-0.84). For cases where fetuses had undergone 1 previous transfusion (n=88) the AUC was 0.76 (95% CI, 0.64-0.88) and at the time of the third cordocentesis for IUT (n=53) it was 0.73 (95% CI, 0.59-0.86). For a 0.59 CFI threshold to diagnosis fetuses with hemoglobin deficit above 5 g/dL, sensitivity values were 87.2%, 88.0%, and 94.1% respectively for fetuses without IUT, with 1 IUT, and with 2 IUTs. Likelihood ratios for positive (LR+) and negative (LR-) test results were 1.98, 2.05, 1.69 and 0.23, 0.21, 0.13 respectively. CONCLUSION: The cardiofemoral index may be an effective noninvasive marker of severe fetal anemia in high-risk fetuses, with accuracy similar for fetuses either with or without previous transfusions.

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The relationship between preeclampsia and the renin-angiotensin system (RAS) is poorly understood. Angiotensin I-converting enzyme (ACE) is a key RAS component and plays an important role in blood pressure homeostasis by generating angiotensin II (Ang II) and inactivating the vasodilator angiotensin-(1-7) (Ang-(1-7)). ACE (I/D) polymorphism is characterized by the insertion (I) or deletion (D) of a 287-bp fragment, leading to changes in ACE activity. In the present study, ACE (I/D) polymorphism was correlated with plasma Ang-(1-7) levels and several RAS components in both preeclamptic (N = 20) and normotensive pregnant women (N = 20). The percentage of the ACE DD genotype (60%) in the preeclamptic group was higher than that for the control group (35%); however, this percentage was not statistically significant (Fisher exact test = 2.86, d.f. = 2, P = 0.260). The highest plasma ACE activity was observed in the ACE DD preeclamptic women (58.1 +/- 5.06 vs 27.6 +/- 3.25 nmol Hip-His Leu(-1) min(-1) mL(-1) in DD control patients; P = 0.0005). Plasma renin activity was markedly reduced in preeclampsia (0.81 +/- 0.2 vs 3.43 +/- 0.8 ng Ang I mL plasma(-1) h(-1) in DD normotensive patients; P = 0.0012). A reduced plasma level of Ang-(1-7) was also observed in preeclamptic women (15.6 +/- 1.3 vs 22.7 +/- 2.5 pg/mL in the DD control group; P = 0.0146). In contrast, plasma Ang II levels were unchanged in preeclamptic patients. The selective changes in the RAS described in the present study suggest that the ACE DD genotype may be used as a marker for susceptibility to preeclampsia.

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The relationship between preeclampsia and the renin-angiotensin system (RAS) is poorly understood. Angiotensin I-converting enzyme (ACE) is a key RAS component and plays an important role in blood pressure homeostasis by generating angiotensin II (Ang II) and inactivating the vasodilator angiotensin-(1-7) (Ang-(1-7)). ACE (I/D) polymorphism is characterized by the insertion (I) or deletion (D) of a 287-bp fragment, leading to changes in ACE activity. In the present study, ACE (I/D) polymorphism was correlated with plasma Ang-(1-7) levels and several RAS components in both preeclamptic (N = 20) and normotensive pregnant women (N = 20). The percentage of the ACE DD genotype (60 percent) in the preeclamptic group was higher than that for the control group (35 percent); however, this percentage was not statistically significant (Fisher exact test = 2.86, d.f. = 2, P = 0.260). The highest plasma ACE activity was observed in the ACE DD preeclamptic women (58.1 ± 5.06 vs 27.6 ± 3.25 nmol Hip-His Leu-1 min-1 mL-1 in DD control patients; P = 0.0005). Plasma renin activity was markedly reduced in preeclampsia (0.81 ± 0.2 vs 3.43 ± 0.8 ng Ang I mL plasma-1 h-1 in DD normotensive patients; P = 0.0012). A reduced plasma level of Ang-(1-7) was also observed in preeclamptic women (15.6 ± 1.3 vs 22.7 ± 2.5 pg/mL in the DD control group; P = 0.0146). In contrast, plasma Ang II levels were unchanged in preeclamptic patients. The selective changes in the RAS described in the present study suggest that the ACE DD genotype may be used as a marker for susceptibility to preeclampsia.

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The purpose of this study was to identify prognostic factors and describe the outcome of prenatally detected renal anomalies associated with multiple malformations and chromosomal defects. Forty-one fetuses were included in the analysis. Prenatal ultrasound reports, neonatal records and autopsy information were retrospectively reviewed. Prognostic factors associated with fetal echography and clinical and laboratory findings on admission were studied. Data were analyzed by univariate analysis in which variables associated with adverse outcome were identified by the Chi-square test or Fisher exact test. The abnormalities associated with renal anomalies were divided into three groups: chromosomal defects (21%), previously described syndromes and conditions (24%), and new sporadic conditions (55%). Of 41 children admitted, 30 (76%) died during the perinatal period. The presence of oligohydramnios was significantly associated with an adverse outcome (OR=11, p=0.05). Male gender was a protective factor against death during the perinatal period (OR=0.11, p=0.01). In conclusion, prenatally detected renal anomalies associated with multiple malformations and chromosomal defects had a poor prognosis. The presence of oligohydramnios increased the risk of death, and male gender had a protective role against poor outcome.

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OBJECTIVE: The purpose of this study was to identify predictive factors of fetal urethral obstruction. METHODS: One hundred and forty-eight children with fetal hydronephrosis were admitted, submitted to a systematic protocol and prospectively followed. Possible predictive factors of urethral obstruction associated with fetal echography and clinical findings on admission were studied. The analysis was conducted in two steps. In a univariate analysis, variables associated with urethral obstruction were identified by the chi(2) test or by Fisher's exact test. Then, the variables that were significantly associated with urethral obstruction were included in a multiple logistic regression analysis. RESULTS: After final adjustment by multiple logistic regression analysis, only two variables were identified as independent predictors of fetal urethral obstruction: oligohydramnios (odds ratio, OR = 5, 95% confidence interval, CI, = 1.3-15, p = 0.01) and megacystis (OR = 9, 95% CI = 2.0-40, p = 0.004). The sensitivity and specificity of the combination of both variables were 60 and 98.5%, respectively. CONCLUSIONS: The presence of oligohydramnios and megacystis on prenatal ultrasound is highly predictive of fetal urethral obstruction.

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With the increasing use of obstetric echography fetal hydronephrosis has been reported more frequently. The purpose of this study was to identify prognostic factors associated with adverse outcome, such as renal failure and death, in fetal hydronephrosis. One hundred and forty-eight children with fetal hydronephrosis were admitted, submitted to a systematic protocol, and prospectively followed. Prognostic factors associated with fetal echography and clinical and laboratory findings on admission were studied. The median follow-up was 39 months. The analysis was conducted in two steps. In a univariate analysis, variables associated with adverse outcome were identified by the Kaplan-Meier method. The variables that were significantly associated with adverse outcome were then included in a multivariate analysis. This analysis, using the multivariate Cox's model, was performed to identify variables that were independently associated with a worse prognosis. Only variables that remained independently associated with adverse outcome were included in the final model. After final adjustment by Cox's multivariate model, three variables were identified as independent predictors of adverse outcome: oligohydramnios, prematurity, and glomerular filtration rate lower than 20 ml/min. Thus, in the presence of oligohydramnios, prematurity, and abnormal renal function, the medical team must plan appropriate follow-up for infants at health centers prepared to investigate and treat uropathies in newborns.

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OBJECTIVE: Fetal hydronephrosis is being detected with increasing frequency and probably this observation will increase as the quality of ultrasound equipment improves. This study evaluated etiology and postnatal clinical outcomes of prenatally detected hydronephrosis. METHODS: In a period of 13 years 148 infants were referred with fetal hydronephrosis. The initial evaluation, after prophylaxis, included ultrasound and voiding cystography. Sequential evaluation was determined by initial findings. RESULTS: Postnatal predominant diagnosis were pelviureteric junction obstruction (24%) and hydronephrosis without obstruction (18%). Follow up median was 38.7 months. Renal function deteriorated in 13 (8.8%) children and 11 (7.4%) died during follow up. Bad prognosis was associated with urethral obstruction. CONCLUSIONS: Fetal hydronephrosis is a clue of urinary tract anomalies. Urinary tract infections and delay in postnatal diagnosis must be prevented in children with this uropathy.

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A case of non-immunitary hydrops fetalis, following supraventricular tachycardia, in a fetus of 29 weeks of a 28-year-old woman at the first pregnancy, who evolved with systemic hypertension and amniorrhexis, being submitted to cesarean deliver. The child needed bilateral thoracic drainage and oro-tracheal intubation for treatment of respiratory failure. Treated with digoxin and diuretic, the newborn went home at the 56th day of life. The authors emphasize the importance of the intrauterine diagnosis of the arrhythmia for the success of the treatment.

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PIP: Leukorrhea constitutes one of the most frequent complaints by women visiting out-patient gynecological clinics. The most common etiological agents are Gardnerella vaginalis, Trichomonas vaginalis, Neisseria gonorrhoea and Candida albicans. Some authors have been able to verify an increased presence of certain pathogenic germs in the vaginal flora for users of contraceptive methods, e.g., for users of IUD, Kivijarvi et al. demonstrated a significantly increased presence of Gardnerella vaginalis considered responsible for a large number of inflammatory diseases of the pelvis in woman. In order to establish the frequency of colpo-cervicitis in users of contraceptive methods a study was initiated of women attending the Family Planning Clinic of the Department of Medicine at UFMG during the period January-December 1986. 305 cytological smears obtained from 111 users of oral hormonal contraceptives and from 194 users of IUD were studied by using the Papanicolaou staining method. A microscopic examination enabling the detection of cells that indicate the presence of Gardnerella vaginalis, trichomonas, Candida, papilloma, and dysplastic cells was performed. The results are listed in two tables. In 34.5% of cases a significant association was established between users of IUD and cells indicating the presence of Gardnerella vaginalis. For users of oral contraceptives that figure was lower, 28.8%. This circumstance has been attributed to probable epithelial lesions caused by incorrect insertion of the IUD and more abundant menstrual flux provoked b the IUD in combination with the hemophilic characteristics of Gardnerella vaginalis. A significant prevalence of papilloma and trichomonas was also noted for women using IUD as opposed to those oral contraceptives. The results demonstrate the necessity of frequent clinical attendance and laboratory tests for women using contraceptive methods, particularly the IUD.^ieng

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PIP: The insertion of intrauterine devices (IUDs) in nulliparous women is a controversial subject, with no unanimous opinion on whether or not to use this method. A study was undertaken to analyze retrospectively the complications of the TCu-200 IUD in nulliparous women compared to complications encountered in multiparous women using the same device. In 1982 and 1983, the Family Planning and Human Reproduction Clinic of the UFMG Medical School in Belo Horizonte, Brazil, inserted 114 IUDs in nulliparas. The principle characteristics of the study group were a low socioeconomic condition, regularity of menstrual flow, and absence of vaginal infection at the time of examination. The choice of method was the spontaneous decision of the women. It was noted that nulliparas requested IUD insertion after previous unsatisfactory experience with other contraceptive methods. 56% of the group were single and 38% married. For comparison, 300 records of multiparous TCU-200 IUD users were studied retrospectively. All insertions were made by the same clinic. The racial, social, and economical characteristics were typical of the users of any free family planning service. Among early complications, pain and lipothymia were encountered at an elevated incidence in nulliparous women when compared to multiparas (p0.01). Among late complications, the appearance of vaginal discharge and metrorrhagia were significant when compared to multiparas (p0.05). Pelvic infection and dysmenorrhea, although more frequent in nulliparas, were not statistically significant. The authors concluded that the IUD should not be used as the contraceptive method of 1st choice in nulliparous women, using it only in exceptional situations.^ieng

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Em 41 gestantes portadoras de hipertensao cronica, correlacionou-se a medida de pressao arterial media (PAM) no segundo trimestre com as ocorrencias do periodo neonatal e com as complicacoes gestacionais apresentadas nestas pacientes. Encontrou-se uma relacao direta da PAM acima de 130 mmHg no segundo trimestre e elevada mortalidade perinatal, alta incidencia de baixo peso e prematuridade. Neste grupo de gestantes ocorreram frequentes complicacoes no pos-parto. Concluiu-se que a PAM no segundo trimestre tem grande valor no prognostico da sobrevivencia do recem-nascido de gestante hipertensa, e ainda correlacionou-se significativamente com a incidencia de complicacoes maternas no periodo pos-parto.

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