A germline chimeric KANK1-DMRT1 transcript derived from a complex structural variant is associated with a congenital heart defect segregating across five generations.

Structural variants (SVs) pose a challenge to detect and interpret, but their study provides novel biological insights and molecular diagnosis underlying rare diseases. The aim of this study was to resolve a 9p24 rearrangement segregating in a family through five generations with a congenital heart defect (congenital pulmonary and aortic valvular stenosis and pulmonary artery stenosis), by applying a combined genomic analysis. The analysis involved multiple techniques, including karyotype, chromosomal microarray analysis (CMA), FISH, genome sequencing (GS), RNA-seq, and optical genome mapping (OGM). A complex 9p24 SV was hinted at by CMA results, showing three interspersed duplicated segments. Combined GS and OGM analyses revealed that the 9p24 duplications constitute a complex SV, on which a set of breakpoints matches the boundaries of the CMA duplicated sequences. The proposed structure for this complex rearrangement implies three duplications associated with an inversion of ~ 2 Mb region on chromosome 9 and a SINE element insertion at the more distal breakpoint. Interestingly, this genomic structure of rearrangement forms a chimeric transcript of the KANK1/DMRT1 loci, which was confirmed by both RNA-seq and Sanger sequencing on blood samples from 9p24 rearrangement carriers. Altogether with breakpoint amplification and FISH analysis, this combined approach allowed a deep characterization of this complex rearrangement. Although the genotype-phenotype correlation remains elusive from the molecular mechanism point of view, this study identified a large genomic rearrangement at 9p24 segregating with a familial congenital heart defect, revealing a genetic biomarker that was successfully applied for embryo selection, changing the reproductive perspective of affected individuals.

A germline chimeric KANK1-DMRT1 transcript derived from a complex structural variant is associated with a congenital heart defect segregating across five generations.

Structural variants (SVs) pose a challenge to detect and interpret, but their study provides novel biological insights and molecular diagnosis underlying rare diseases. The aim of this study was to resolve a 9p24 rearrangement segregating in a family through five generations with a congenital heart defect (congenital pulmonary and aortic valvular stenosis, and pulmonary artery stenosis), by applying a combined genomic analysis. The analysis involved multiple techniques, including karyotype, chromosomal microarray analysis (CMA), FISH, whole-genome sequencing (WGS), RNA-seq and optical genome mapping (OGM). A complex 9p24 SV was hinted at by CMA results, showing three interspersed duplicated segments. Combined WGS and OGM analyses revealed that the 9p24 duplications constitute a complex SV, on which a set of breakpoints match the boundaries of the CMA duplicated sequences. The proposed structure for this complex rearrangement implies three duplications associated with an inversion of ~ 2Mb region on chromosome 9 with a SINE element insertion at the more distal breakpoint. Interestingly, this hypothesized genomic structure of rearrangement forms a chimeric transcript of the KANK1/DMRT1 loci, which was confirmed by RNA-seq on blood from 9p24 rearrangement carriers. Altogether with breakpoint amplification and FISH analysis, this combined approach allowed a deep characterization of this complex rearrangement. Although the genotype-phenotype correlation remains elusive from the molecular mechanism point of view, this study identified a large genomic rearrangement at 9p segregating with a familial congenital clinical trait, revealing a genetic biomarker that was successfully applied for embryo selection, changing the reproductive perspective of affected individuals.

Índice de rendimiento biventricular del corazón fetal

Objetivo. Proponer un índice de rendimiento biventricular basado en los valores promedio de la duración del ciclo cardíaco de cada uno de los ventrículos, determinar la variable incluida en el estudio con la correlación estadística más significativa, establecer valores de referencia que permitan identificar el trabajo de cada ventrículo en función de dicha variable y obtener un índice de gasto biventricular equilibrado. Metodología. Estudio prospectivo y transversal en fetos de 168 gestantes, en embarazos entre las 16 y 38 semanas sin patologías materno-fetales. Se obtuvieron ondas de velocidad de flujo de ambas válvulas atrioventriculares y el tiempo total del ciclo sístole-diástole se calculó en milisegundos para cada válvula. Se calcularon promedios, desviación estándar y puntuación Z del tiempo sistólico-diastólico para cada ventrículo y el índice de rendimiento ventricular individual dividiendo el valor obtenido entre la frecuencia cardiaca fetal. Se obtuvo el valor promedio de ambos y este, al ser dividido por la frecuencia cardíaca, permitió obtener el índice de rendimiento biventricular para establecer la correlación entre este, la frecuencia cardiaca fetal y la edad de gestación. Resultados. Se halló valores de tiempo sistólico-diastólico en milisegundos para el ventrículo derecho de 420,8 (DE ±28,3) y para el ventrículo izquierdo de 418,8 (DE ±26,3), sin diferencias estadísticamente significativas (p=0,371). La correlación con la frecuencia cardíaca fetal resultó negativa para ambos ventrículos (-0,491 y -0,553; p<0,05). El tiempo promedio biventricular fue de 418,37 ms (± 20,59) y la correlación con la edad gestacional de 0,48 (p<0,05); la correlación con la frecuencia cardiaca fetal fue negativa, -0,50 (p<0,05). El índice de rendimiento biventricular mostró valores de 2,8 (extremos 2,4 (P5) y 3,4 (P95)). La correlación entre el índice de rendimiento biventricular y la frecuencia cardiaca fetal fue 0,78 (p<0,05), de menor grado (0,27) con la edad gestacional. Conclusiones. Se demostró que los tiempos sistólico-diastólicos de cada ventrículo no difirieron entre sí y se correlacionaron de manera negativa con la frecuencia cardiaca fetal. Se comprobó que es posible evaluar el ciclo cardíaco fetal de cada ventrículo mediante el índice de rendimiento ventricular, así como calificar con el índice de rendimiento biventricular el gasto cardíaco combinado como equilibrado.

Objectives: To propose a biventricular performance index based on the average values of the duration of the cardiac cycle of each of the ventricles, to determine the variable included in the study with the most significant statistical correlation, to establish reference values that allow the work of each ventricle to be identified according to this variable, and to obtain a balanced biventricular output index. Methodology: Prospective and cross-sectional study in fetuses of 168 pregnant women, in pregnancies between 16 and 38 weeks without maternal-fetal pathologies. Flow velocity waves were obtained from both atrioventricular valves and the total systole-diastole cycle time was calculated in milliseconds for each valve. Averages, standard deviation, and Z-score were calculated of the systolic-diastolic time for each ventricle and the individual ventricular performance index (VPI) were calculated by dividing the value obtained by the fetal heart rate (FHR). The average value of both was obtained and this, when divided by the heart rate, made it possible to obtain the biventricular performance index (BPI) to establish the correlation between this, the fetal heart rate and gestational age. Results: Systolic-diastolic time values in milliseconds for the right ventricle were 420.8 (SD ±28.3) and for the left ventricle 418.8 (SD ±26.3), with no statistically significant differences (p<0.371). The correlation with the FHR was negative for both ventricles: (-0.491 and -0.553; p<0.05). The mean biventricular time was 418.37 ms (±20.59) and the correlation with gestational age was 0.48 (p<0.05); the correlation with FHR was negative, -0.50 (p<0.05).The BPI showed values of 2.8 (extremes 2.4 (P5) and 3.4 (P95)). The correlation between BPI and FHR was 0.78 (p<0.05) and of lesser degree with gestational age (0.27; p<0.05). Conclusions: It was demonstrated that the systolic-diastolic times of each ventricle did not differ from each other and were negatively correlated with fetal heart rate. It was shown that it is possible to evaluate the fetal cardiac cycle of each ventricle by means of the ventricular performance index as well as to qualify with the biventricular performance index the combined cardiac output as balanced.

Maternal ingestion of cocoa causes constriction of fetal ductus arteriosus in rats.

Maternal consumption of polyphenol-rich foods has been associated with fetal ductus arteriosus constriction (DAC), but safety of chocolate exposure in fetal life has not been studied. This experimental study tested the hypothesis that maternal cocoa consumption in late pregnancy causes fetal DAC, with possible associated antioxidant effects. Pregnant Wistar rats, at the 21st gestational day, received by orogastric tube cocoa (720 mg/Kg) for 12 h, indomethacin (10 mg/Kg), for 8 h, or only water, before cesaren section. Immediately after withdrawal, every thorax was obtained and tissues were fixed and stained for histological analysis. The ratio of the narrowest part of the pulmonary artery to the fetal ductus inner diameter and increased ductal inner wall thickness characterized ductal constriction. Substances reactive to thiobarbituric acid were quantified. Statistical analysis used ANOVA and Tukey test. Cocoa (n = 33) and indomethacin (n = 7) reduced fetal internal ductus diameter when compared to control (water, n = 25) (p < 0.001) and cocoa alone increased ductus wall thickness (p < 0.001), but no change was noted in enzymes activity. This pharmacological study shows supporting evidences that there is a cause and effect relationship between maternal consumption of cocoa and fetal ductus arteriosus constriction. Habitual widespread use of chocolate during gestation could account for undetected ductus constriction and its potentially severe consequences, such as perinatal pulmonary hypertension, cardiac failure and even death. For this reason, dietary guidance in late pregnancy to avoid high chocolate intake, to prevent fetal ductal constriction, may represent the main translational aspect of this study.

Development of the Z-score for the measurement of myocardial thickness by two-dimensional echocardiography in normal fetuses.

BACKGROUND: In the current literature, there is a tendency to describe normal values of echocardiographic measurements by means of the Z-score. In fetal cardiology, these Z-score equations are still being established. Measurement of myocardial thickness is an important assessment, especially in fetuses of diabetic mothers, because of the risk of developing myocardial hypertrophy secondary to elevated maternal blood glucose levels. OBJECTIVE: To determine the percentiles and to develop the Z-score equations of right and left ventricular lateral walls and interventricular septum measurements using two-dimensional echocardiography in normal fetuses between 24 and 34 weeks of gestation. METHODS: This is a prospective cross-sectional study that was performed in single fetuses with normal heart from nondiabetic pregnant women. Measurements of the lateral walls of the right and left ventricles and the interventricular septum were made. RESULTS: Eight hundred and seventy three pregnant women were included. We determined the percentiles of the measurements for each gestational age. The Z-score equation was developed for each of the measurements: right ventricular lateral wall measurement [RVLW = x-(-1 + 0.109 * GA)/0.4], left ventricle lateral wall measurement [LVLW = x-(-1.366 + 0.12 * GA)/0.43], and interventricular septum, both at the four-chamber view [IVS4ch = (x-(-1.113 + 0.107 * GA)/0.4] and at the left ventricular outflow tract plane [IVSLVOT = (x-(-0.581 + 0.084 * GA)/0.35]. CONCLUSION: The present study allowed the demonstration of the percentiles and the Z-score equations for each of the measurements studied.

Brazilian fetal cardiology guidelines ­ 2019 / Diretriz brasileira de cardiologia fetal ­ 2019

Over the years, Fetal Cardiology have been incorporated into the daily practice of Pediatric Cardiology. What was once restricted to a few fetal heart researchers, has slowly been incorporated into health institutions that deal with congenital heart diseases (CHD). Fetal echocardiography has generated extensive knowledge of the natural and modified history of heart diseases in utero, and normal fetal heart physiology and anatomy. The benefits of fetal diagnosis have become unquestionable over the years. Pioneers in the area succeeded in demystifying the fetal heart examination and proving the importance of screening for cardiac abnormalities during obstetric examinations. Prenatal detection rates have increased, and interest in fetal echocardiography is, thus, no longer merely a diagnostic tool; it has gone on to become a tool of the utmost importance in assisting medical and, progressively, interventional treatment of specific anomalies that occur in fetal life. A vast body of literature currently supports the practice of Fetal Cardiology. In addition to diagnosis, anatomical and functional particularities may be identified in utero, with implications on the delivery planning and pre and postnatal management. Prenatal diagnosis has certainly led to increase the number of babies with complex heart diseases in Pediatric Cardiology hospital beds. Prior to this, children with complex heart diseases did not survive the immediate neonatal period and died in neonatal intensive care units without being diagnosed. Nowadays, these children require increasingly careful and specific management involving Pediatric Cardiology and thus modifying the practice of Neonatal Cardiology. Despite the vast literature pertinent to Fetal Cardiology, due to the restricted number of cases, there is a lack of studies with large populations and randomization processes, being the information based on observational studies and description of small samples or cases reports. However, the accumulated knowledge is already enough to develop scientific statements or guidelines. In April 2014, the American Heart Association (AHA) published the first scientific statement for Fetal Cardiology, encompassing all the practical aspects involved in this area, including screening, diagnosis, medical or interventional therapy, counseling, delivery planning, and neonatal treatment. Considering this extremely thorough and highly useful document, we have accepted the challenge of bringing together professionals dedicated to Fetal Cardiology from different regions of Brazil in order to jointly establish guidelines which are adapted to our reality and which also take into consideration knowledge created in Brazil. We believe that the information brought together in this document will be of great use to professionals who face the challenge of dealing with possible abnormalities that affect the fetal heart in their daily practice. (AU)

Pulmonary Vein Flow Impedance: An Early Predictor of Cardiac Dysfunction in Intrauterine Growth Restriction.

INTRODUCTION: In intrauterine growth restriction (IUGR), increased uteroplacental vascular impedance contributes to preferential flow to left ventricle (LV), with consequent alteration of its compliance and increased left atrial (LA) pressure. Pulmonary vein pulsatility index (PVPI) reflects the increased impedance to LA filling and could be used as a cardiac monitoring parameter in IUGR. MATERIAL AND METHODS: A total of 27 IUGR fetuses (group 1), 28 fetuses with appropriate growth for gestational age from hypertensive mothers (group 2), and 28 controls (group 3) were studied. Pulsatility indices (PIs) of pulmonary veins and ductus venosus were calculated by Doppler echocardiography. Obstetric ultrasound was used to assess the PIs of uterine, umbilical, and middle cerebral arteries. Statistical analysis used analysis of variance, post-hoc Tukey, and Pearson's tests. RESULTS: Mean PVPI was higher in IUGR group (1.27 ± 0.39) when compared to groups 2 (1.02 ± 0.37; p = 0.01) and 3 (0.75 ± 0.12; p < 0.001). In group 2, moderate correlation between PVPI and ductus venosus pulsatility index (DVPI) was found but not between PVPI and cerebroplacental ratio (CPR). DISCUSSION: Higher PVPI in IUGR reflects decreased LV compliance and altered LA dynamics. As LV dysfunction precedes right ventricle, our results suggest that PVPI could be an early echocardiographic parameter of fetal diastolic function in IUGR.

Corrigendum to "Behaviour of the Foramen Ovale Flow in Fetuses with Intrauterine Growth Restriction".

[This corrects the article DOI: 10.1155/2018/1496903.].

Behaviour of the Foramen Ovale Flow in Fetuses with Intrauterine Growth Restriction.

Background: Foramen ovale (FO) flow may be altered in IUGR. This study was designed to test this hypothesis. Methods: Forty pregnant women (24-38 weeks) were divided into 3 groups: group I (IUGR), group II (adequate growth and maternal hypertension), and group III (normal controls). Impedance across the FO was assessed by the FO pulsatility index (FOPI): (systolic velocity - presystolic velocity)/mean velocity. Statistical analysis utilized ANOVA, Tukey test, and ROC curves. Results: Mean FOPI in IUGR fetuses (n = 15) was 3.70 ± 0.99 (3.15-4.26); in the group II (n = 12), it was 2.84 ± 0.69 (2.40-3.28), and in the group III (n = 13), it was 2.77 ± 0.44 (2.50-3.04) (p=0.004). FOPI and UtA RI were correlated (r = 0.375, p=0.017), as well as FOPI and UA RI (r = 0.356, p=0.024) and, inversely, FOPI and MCA RI (r = -0.359, p=0.023). Conclusions: The FO flow pulsatility index is increased in fetuses with IUGR, probably as a result of impaired left ventricular diastolic function.

Polyphenol-rich food general and on pregnancy effects: a review.

This review aimed to investigate possible protective or deleterious effects of polyphenol-rich foods (PRF) on chronic diseases, e.g. cardiovascular, and in pregnant women, along with their antioxidant and anti-inflammatory action. A great variety of foods and beverages, such as herbal teas, grape and orange derivatives, dark chocolate, and many others contain high concentrations of flavonoids and are freely consumed by the general population. In humans, PRF consumption reduces lipid peroxidation, and several studies have shown a positive correlation between an increased consumption of PRF and a decrease in the incidence of cardiovascular disease. On the other hand, current studies have suggested that maternal ingestion of PRF, especially during the third trimester of pregnancy, could be associated to fetal ductal constriction (DC). Fetuses exposed to this type of diet show higher ductal velocities and lower pulsatility indexes, as well as larger right ventricles than those exposed to minimal amounts of these substances. The underlying mechanism involved in these conditions has not been entirely elucidated, but it seems to be a result of the antioxidant and anti-inflammatory effects of polyphenols by some pathway. Furthermore, taking into account the deleterious effect in late-pregnancy against the numerous positive effects associated to polyphenols, this dual behavior deserves attention particularly to control the dietary ingestion of PRF during gestation. In this line, same PRF, natural constituents of human diet, may represent risk to fetal in late pregnancy compared to the use of nonsteroidal anti-inflammatory drugs.

Dynamics of pulmonary venous flow in fetuses with intrauterine growth restriction.

OBJECTIVE: To test the hypothesis that the pulmonary vein pulsatility index (PVPI) is higher in fetuses with growth restriction (IUGR) than in normal fetuses. METHODS: Twenty-two fetuses with IUGR and twenty-one (21) fetuses with appropriate growth for gestational age from healthy mothers were studied. PVPI was calculated by Doppler echocardiography [maximal velocity (systolic or diastolic peak) - pre-systolic peak / mean velocity]. Obstetric ultrasound was used to assess fetal biometry and Doppler to assess the uterine, umbilical and middle cerebral arteries PI. Statistical analysis used t test and Pearson's correlation. RESULTS: Mean gestational age was 31.5 +/- 2.1 weeks in the control group and 31.4 +/- 3.1 weeks in IUGR (P = 0.91). The PI of uterine and umbilical arteries were higher in IUGR than in controls (P < 0.001). Mean PVPI in IUGR fetuses was 1.31 +/- 0.41, and in controls it was 0.83 +/- 0.11 (P < 0.001). CONCLUSION: The pulsatility index of pulmonary venous flow in fetuses with growth restriction is higher than in normal fetuses, probably as a result of left atrial dynamics alteration secondary or not to fetal left ventricular diastolic dysfunction. © 2014 John Wiley & Sons, Ltd.

Development and validation of a food frequency questionnaire for consumption of polyphenol-rich foods in pregnant women.

Previous studies have shown that maternal consumption of polyphenol-rich foods after the third trimester of pregnancy may interfere with the anatomical and functional activity of the fetal heart as, to our knowledge, there are no validated instruments to quantify total polyphenols in pregnant women. The aim of this study was evaluate the reproducibility and validity of a food frequency questionnaire (FFQ), with 52 items, to assess the intake of polyphenol-rich foods in pregnant women in Brazil. This cross-sectional study included 120 pregnant women who participated in nutritional interviews in two moments. The intake of polyphenols estimated by the developed FFQ was compared with the average of two 24-h recalls (24HR), with the average intake measured by a 3-day food diary (D3days) and with the urinary excretion of total polyphenols. The triangular method was applied to calculate Pearson's correlation coefficients, intraclass correlation and Bland-Altman plots for the FFQ, using an independent biochemical marker, in addition to classification by quarters of consumption. The questionnaires were log transformed, adjusted for body mass index and gestational age. The adjustment for energy was applied only of 24HR and D3days. Analysis of the reproducibility between the FFQ showed a very high correlation (r = 0.72; P < 0.05). A low but significant association was observed between the FFQ and urinary excretion (0.23; P = 0.01). The association between the dietary survey methods was moderate to very high (r = 0.36 to r = 0.72; P < 0.001). In conclusion, this questionnaire showed reproducibility and validity for the quantification of consumption of total polyphenols in pregnant women.

Nitric oxide and reactive species are modulated in the polyphenol-induced ductus arteriosus constriction in pregnant sheep.

OBJECTIVE: Because we have previously demonstrated the relation between polyphenol-rich foods (PRF) consumption and ductus arteriosus constriction, in this work, pregnant sheep were submitted to oral PRF intake for 14 days to understand how this process occurs. Fetal Doppler echocardiography, oxidative and inflammatory biomarkers and total polyphenol excretion were evaluated. RESULTS: The high polyphenol intake induced ductus arteriosus constriction by 71.6% increase in systolic (P = 0.001) and 57.8% in diastolic velocities (P = 0.002), and 18.9% decrease in pulsatility index (P = 0.033), along with 1.7-fold increase in total polyphenol excretion, 2.3-fold decrease in inflammatory mediator nitric oxide and following redox status changes (mean ± standard deviation): higher protein carbonyls (1.09 ± 0.09 and 1.49 ± 0.31), catalase (0.69 ± 0.39 and 1.44 ± 0.33) and glutathione peroxidase (37.23 ± 11.19 and 62.96 ± 15.03) in addition to lower lipid damage (17.22 ± 2.05 and 12.53 ± 2.11) and nonprotein thiols (0.11 ± 0.04 and 0.04 ± 0.01) found before and after treatment, respectively. Ductal parameters correlated to NOx , catalase, glutathione peroxidase and protein carbonyl. CONCLUSION: Our results highlight the need to reduce maternal PRF intake in late pregnancy to prevent fetal duct constriction through NO-mediated vasoconstrictive action of polyphenols.

Maternal-fetal attachment and prenatal diagnosis of heart disease.

OBJECTIVE: To test the hypothesis that there are differences in the level of maternal-fetal attachment before and after fetal echocardiography in the presence or absence of cardiac abnormalities. STUDY DESIGN: Cohort study in which the mothers responded to a validated Maternal-Fetal Attachment Scale. The study compared a group of pregnant women with diagnosis of fetal heart disease (FHD) with a group without this diagnosis ("no fetal heart disease" - NFHD). RESULTS: 197 pregnant women were included, 96 FHD and 101 NFHD. Maternal-fetal attachment at the initial and final periods showed no significant baseline differences between groups (p=0.081). At the final period, migration from medium to high level of attachment was significantly higher in FHD (p=0.017). Transition from medium to high levels comparing the initial and final periods was more pronounced in FHD (p=0.009). CONCLUSION: Diagnosis of fetal heart disease increases the level of maternal-fetal attachment.

Myocardial velocities, dynamics of the septum primum, and placental dysfunction in fetuses with growth restriction.

INTRODUCTION: Diastolic dysfunction may occur in fetuses with intrauterine growth restriction (IUGR) and may be assessed by myocardial tissue Doppler (MTD). We previously have shown that excursion index of the septum primum (EISP) is reduced in IUGR fetuses over 30 weeks because of a higher left atrial pressure. PATIENTS, SETTING, AND DESIGN: The sample was made up of 14 fetuses with IUGR. MTD examination was carried out with the sample volume placed at the basal lateral wall of the left ventricle (LV), interventricular septum (IVS), and free wall of the right ventricle (RV) to determine E'/A' ratios. EISP was calculated as the ratio between the maximal excursion of the septum primum into the left atrium during diastole and the maximal diastolic diameter of the left atrium. Mitral and tricuspid flows were assessed by the conventional Doppler method. OUTCOME MEASURES: Pearson's correlation test was used to analyze the correlations between the parameters. RESULTS: A positive correlation was observed between UARI and E'/A' ratios for RV (r = 0.63, P = .02), IVS (r = 0.59, P = .03), and LV (r = 0.41, P = .15). There was a negative correlation between EISP and IVS E'/A' ratios (r = -0.58, P = .03), and a positive correlation for LV (r = 0.49, P = .08). At the RV position, a weak negative correlation was observed (r = -0.32, P = .26). CONCLUSIONS: A higher left atrial pressure in fetuses with IUGR, indicated by the lower mobility of the septum primum, is accompanied by higher ratios between early and late diastolic myocardial velocities. Placental dysfunction was correlated to septal E'/A' ratios. Fetal MTD can be a useful method to assess severity of placental dysfunction and fetal distress.

In vivo toxicological evaluation of polymeric nanocapsules after intradermal administration.

Polymeric nanocarriers have shown great promise as delivery systems. An alternative strategy has been to explore new delivery routes, such as intradermal (i.d.), that can be used for vaccines and patch-based drug delivery. Despite their many advantages, there are few toxicity studies, especially in vivo. We report a safety assessment of biodegradable poly(ɛ-caprolactone) lipid-core nanocapsules (LNC) with a mean size of 245±10nm following single and repeated intradermal injections to Wistar rats. Suspensions were prepared by interfacial deposition of polymer. The animals (n=6/group) received a single-dose of saline solution (1.2ml/kg) or LNC (7.2×10(12)LNC/kg), or repeated-doses of two controls, saline solution or Tween 80 (0.9ml/kg), or three different concentrations of LNC (1.8, 3.6, and 5.4×10(12)LNC/kg) for 28 consecutive days. Clinical and physiological signs and mortality were observed. Samples of urine, blood, and tissue were used to perform toxicological evaluation. There were no clinical signs of toxicity or mortality, but there was a slight decrease in the relative body weights in the Tween 80-treated group (p<0.01) after repeated administration. No histopathological alterations were observed in tissues or significant changes in blood and urinary biomarkers for tissue damage. Mild alterations in white blood cells count with increases in granulocytes in the Tween-80 group (p<0.05) were found. Genotoxicity was evaluated through the comet assay, and no statistical difference was observed among the groups. Therefore, we conclude that, under the conditions of these experiments, biodegradable LNC did not present appreciable toxicity after 28 consecutive days of intradermal administration and is promising for its future application in vaccines and patch-based devices for enhancing the delivery of drugs.

Deleterious effects of maternal ingestion of cocoa upon fetal ductus arteriosus in late pregnancy.

Cocoa powder has twice more antioxidants than red wine and three times more than green tea. Ten percent of its weight is made up of flavonoids. Cocoa has antioxidant and anti-inflammatory effects by downregulating cyclooxigenase-2 receptors expression in the endothelium and enhancing nitric oxide bioavailability. There are evidences that while polyphenols ingestion have cardioprotective effects in the adult, it may have deleterious effect on the fetus if ingested by the mother on the third trimester of pregnancy, causing intrauterine fetal ductus arteriosus (DA) constriction. Polyphenols present in many foods and their anti-inflammatory and antinociceptive activities have been shown to be as or more powerful than those of indomethacin. These effects are dependent on the inhibition of modulation of the arachidonic acid and the synthesis of prostaglandins, especially E-2, which is responsible for fetal DA patency. So, we hypothesized that this same mechanism is responsible for the harmful effect of polyphenol-rich foods, such as cocoa, upon the fetal DA after maternal intake of such substances in the third trimester of pregnancy, thereby rising the perspective of a note of caution for pregnant women diet.