Women's multisite microbial modulation during pregnancy.

The composition of female microbiome varies with age, physiological and socio-behavior conditions. Also, changes in microbiome composition are observed as pregnancy progresses, especially in the vaginal site. Together with the physiological adaptations of gestation, changes in microbiome composition seem to be fundamental for proper fetal development. This study aimed at simultaneously evaluating the vaginal, gut, and oral microbiome of healthy pregnant women, and comparing it with those observed in healthy non-pregnant women of reproductive age. In a cross-sectional study, vaginal, oral and gut samples were collected from 42 pregnant and 18 non-pregnant women, and the microbiome composition was evaluated by 16S rRNA sequencing, using Illumina platform. In the pregnant group, we observed a positive correlation between Eubacterium and Akkermansia in the gut samples; between Eubacterium and Ruminococcus in the vaginal samples; and between Streptococcus and Gemella in the oral samples. Notwithstanding, we observed a negative correlation between Lactobacillus and Atopobium and between Lactobacillus and Gardnerella in vaginal microbiome. Prevotella was the only genus found in all three sites studied; however, there was no signal of bacterial influence between sites during pregnancy. These results suggest that in addition to hormonal and immunological variations during healthy pregnancy, the female body also undergoes microbiome modulation in multiple sites in order to maintain an eubiotic status.

Unveiling the pathophysiology of gestational diabetes: Studies on local and peripheral immune cells.

Gestational diabetes mellitus (GDM) has been associated with impaired maternal immune response. Our aim was to review the available literature linking immune cells profile to GDM, in order to comprehend the role that different subpopulations play in the development of this pathology. We searched in PubMed for studies published in the last decade on circulating levels and placenta expression of immune cells on GDM. We identified 18 studies with several differences regarding the study design, clinical characteristics, number of participants, cell subpopulation and type of sample. Most studies assessed only one subpopulation either in peripheral blood or placenta and did not analyse functional properties of the cells. The most frequently evaluated immune cells were T lymphocytes, especially regulatory T (Tregs), and natural killer (NK) cells in the peripheral blood, and placental macrophages. No studies analysing B cells were identified, and only one study each evaluating γδT cells, dendritic cell (DC) and monocytes was found. Although there are controversies, at least one study reported positive association between GDM and CD4+ (activated), Tregs, Th17 and γδT cells; neutrophil/lymphocyte; NK cell (cytotoxic); macrophages; and monocytes. The number of studies is still small, so caution should be exercised in interpreting the data, and further research is required to validate these findings and establish the role of adaptive and innate immune cells in GDM pathophysiology.

Versican silencing in BeWo cells and its implication in gestational trophoblastic diseases.

Versican is a proteoglycan known to interact with cells to influence their ability to proliferate, differentiate, migrate, invade and assemble extracellular matrix, with all of these cell functions present during placentation. In the placenta, cytotrophoblast cells have the ability to differentiate into the syncytiotrophoblast, a mechanism that is greatly increased in gestational trophoblastic diseases (GTD). Nevertheless, the molecular signaling underlying the increased syncytiotrophoblast differentiation are still being unveiled and may result in novel therapeutic targets for GTD. Versican expression was investigated to establish its differential expression among GTD (partial moles, complete moles, invasive moles and choriocarcinoma) and the possible functional outcomes from versican gene silencing. Tissue samples had their versican expression evaluated using immunohistochemistry and RT-PCR. BeWo cells were employed for versican silencing with siRNA and the efficiency was confirmed by RT-PCR, immunofluorescence and flow cytometry. Cell death and forskolin-induced syncytialization were analyzed by a morphological analysis and human chorionic gonadotropin (hCG) production using immunofluorescence. Versican V0 and V1 isoforms were mainly expressed in the syncytiotrophoblast and they were the most expressed in benign rather than in malignant tumors. BeWo cells also expressed V0 and V1 isoforms, but only in cells undergoing syncytial fusion. After versican silencing, cell death was greatly increased, whereas spontaneous and forskolin-induced syncytialization decreased as well as hCG production. Versican is differentially expressed in GTD and is important for hydatidiform moles pathophysiology, protecting trophoblast cells from death and playing a role in their differentiation and functionality.

Monocyte profile in peripheral blood of gestational diabetes mellitus patients.

OBJECTIVE: Gestational diabetes Mellitus has been considered an inflammatory disease involving different cells and mediators in its development. The role of innate immune cells in GDM physiopathology remains unclear, therefore this study was conducted to assess monocyte profile in GDM patients. DESIGN: This was a case-control study including 20 glucose-tolerant pregnant women (controls) and 18 GDM patients. METHODS: Flow cytometry was used to assess peripheral blood monocytes subsets (classical, intermediate, non-classical), the expression of TLR4 and CCR2 chemokine receptor (CD192) and cytokines (TNFA, IL6, IL10) secretion by monocytes subsets. In addition, sCD14 serum levels were evaluated by ELISA. RESULTS: We observed increased percentage of CD14+ cells, decreased frequency of intermediate monocytes (CD14+CD16+), and lower percentage of circulating monocytes (classical, intermediate and non-classical) that express TLR4 in the diabetic group compared to controls. Soluble CD14+ serum levels were higher in GDM patients compared to controls. There were no differences in the expression of the CCR2 chemokine receptor and cytokines (TNFA, IL6 and IL10) secretion between the studied groups. CONCLUSIONS: Our results demonstrated that GDM patients present impaired monocyte profile in the peripheral blood, suggesting that these cells are involved in GDM physiopathology.

Cervical cerclage placement decreases local levels of proinflammatory cytokines in patients with cervical insufficiency.

BACKGROUND: Cervical insufficiency is characterized by premature, progressive dilation and shortening of the cervix during pregnancy. If left unattended, this can lead to the prolapse and rupture of the amniotic membrane, which usually results in midtrimester pregnancy loss or preterm birth. Previous studies have shown that proinflammatory cytokines such as interleukin-1ß, interleukin-6, interleukin-8, and tumor necrosis factor alpha are up-regulated in normal parturition but are also associated with preterm birth. Studies evaluating such markers in patients with cervical insufficiency have evaluated only their diagnostic potential. Even fewer studies have studied them within the context of cerclage surgery. OBJECTIVES(S): The objective of the study was to evaluate the impact of local and systemic inflammatory markers on the pathogenesis of cervical insufficiency and the effect of cerclage surgery on the local immune microenvironment of women with cervical insufficiency. STUDY DESIGN: We recruited 28 pregnant women (12-20 weeks' gestation) diagnosed with insufficiency and referred for cerclage surgery and 19 gestational age-matched normal pregnant women as controls. Serum and cervicovaginal fluid samples were collected before and after cerclage surgery and during a routine checkup for normal women and analyzed using a targeted 13-plex proinflammatory cytokine assay. RESULTS: Before surgery, patients with cervical insufficiency had higher levels of interleukin-1ß, interleukin-6, interleukin-12, monocyte chemoattractant protein-1 and tumor necrosis factor alpha in cervicovaginal fluid compared to controls, but after surgery, these differences disappeared. No differences were found in serum of insufficiency versus control women. In patients with insufficiency, the levels of interleukin-1ß, interleukin-6, interleukin-8, monocyte chemoattractant protein-1, and interferon gamma in cervicovaginal fluid declined significantly after cerclage compared with before intervention, but these changes were not detected in serum. CONCLUSION: Compared with normal women, patients with cervical insufficiency have elevated levels of proinflammatory cytokines in cervicovaginal fluid but not in serum, suggesting a dysregulation of the local immune environment. Cerclage intervention led to a significant decline in these proinflammatory cytokines, suggesting that cerclage may help reduce local inflammation in cervical insufficiency.

VEGF 936C/T Polymorphism and Gestational Trophoblastic Neoplasia.

OBJECTIVE: To evaluate the relationship between the 936C/T polymorphism of VEGF and the occurrence of gestational trophoblastic neoplasia (GTN). STUDY DESIGN: A retro- spective study that included 8 patients with complete hydatidiform -mole (CHM) that evolved into spontane- ous remission (SR), 12 pa- tients with CHM that prog- ressed to GTN, and 20 control (C) patients without obstetric complications. Polymorphisms were detected by polymerase chain reaction-amplified technique of patients' DNA, and genotype frequencies were compared between the groups. RESULTS: . The genotype frequencies of the VEGF 936C/T polymorphism were as follows: SR group, 100% CC genotype; GTN group, 50.0% CC, 41.7% CT, and 8.3% TT; C group, 30.0% CC, 65.0% CT, and 5.0% TT. Genotype frequencies did not differ significantly be- tween the SR and GTN groups, although a trend was observed (p=0.06). Genotype frequencies did differ sig- nificantly between the combined group of all patients with CHM (SR+GTN) and the C group (p=0.03). CONCLUSION: This study did not identify a different VEGF 936CT genotype profile for patients with CHM who undergo SR versus those who progress to GTN. However, the, results do suggest that this polymor- phism may affect susceptibil- ity to CHM. Larger groups may improve the results of assessments of the predictive parameters of GTN.

Evaluation of biochemical markers combined with uterine artery Doppler parameters in fetuses with growth restriction: a case-control study.

PURPOSE: Assessing the biochemical markers levels and the uterine artery Doppler (UtA) parameters in fetuses with growth restriction (FGR). METHODS: Prospective case-control study included 66 patients with diagnosis of FGR and 64 healthy pregnancies at 24-41 weeks of gestation. For both groups, maternal circulating concentrations of biochemical factors of soluble fms-like tyrosine kinase-1 (sFlt-1), soluble endoglin(sEng), adiponectin, A disintegrin and metalloproteinases (ADAM-12), pregnancy-associated plasma protein-A (PAPP-A), angiopoietin-2 (ANGI-2), vascular endothelial growth factor (VEGF) and transforming growth factor-ß (TGF-ß) were assayed by ELISA and UtA by Doppler were performed. ANOVA, Mann-Whitney tests and Pearson correlation coefficient were applied to compare the biochemical factors, UtA Doppler and EFW Z-score between the groups. RESULTS: Concentrations of sFlt-1, sEng, PAPP-A were significantly higher in FGR than controls (p < 0.0001, p = 0.02 and p = 0.03, respectively), but concentration of ANGI-2 (p < 0.0001) was significantly lower in FGR than controls and ADAM-12 levels had a tendency to be lower in the FGR, though not statistically significant (p = 0.059). Increased sEng concentrations were correlated with abnormal UtA Doppler in FGR. CONCLUSION: Fetal growth restriction fetuses showed increased serum levels of sFlt-1, sEng and PAPP-A with levels of ANGI-2 decreased and a positive association between elevated concentrations of sEng and changing impedance of UtA Doppler were observed.

Getting too sweet: galectin-1 dysregulation in gestational diabetes mellitus.

Galectin-1 (gal-1) is a prototype carbohydrate-binding protein, whose dysregulation is associated with adverse pregnancy outcomes such as spontaneous abortion and pre-eclampsia. Furthermore, it is known that faulty gal-1 protein production or gene regulation can be caused by single-nucleotide polymorphisms in the LGALS1 gene. Gestational diabetes mellitus (GDM) is also an adverse pregnancy outcome and the most common metabolic disorder during gestation. However, gal-1 expression patterns during GDM remain largely unknown. Our aims were to define local and peripheral gal-1 expression patterns during pregnancy, and to investigate LGALS1 gene polymorphisms in GDM patients. Circulating gal-1 levels were determined by ELISA in GDM patients and normal pregnant controls, and LGALS1 gene polymorphisms were assessed for association with GDM. Placental tissues were collected from control and GDM term pregnancies to evaluate local gal-1 expression by immunofluorescence. Our results show that GDM is associated with a failure to increase circulating gal-1 levels during the second and third trimester, as well as overexpression of gal-1 in placental tissue. Additionally, the LGALS1 polymorphism rs4820294 was associated with the development of GDM. In pregnancies complicated by GDM, we observed gal-1 dysregulation both locally in the placenta and peripherally in the circulation. Furthermore, the association between the LGALS1 polymorphism and GDM may indicate a genetic contribution to this adverse pregnancy outcome.

Decorin and biglycan immunolocalization in non-villous structures of healthy and pathological human placentas.

AIMS: Decorin and biglycan are members of the small leucine-rich proteoglycan family, and constituents of both the extracellular matrix (ECM) and the cell surface. They are recognized as important factors in the control of proliferation, migration and invasion in vivo and in vitro. In this study, the localization patterns of decorin and biglycan were determined in healthy placentas and in highly invasive placental pathologies. METHODS AND RESULTS: The study included immunolocalization of decorin and biglycan in samples of first-trimester and term placentas, placenta accreta, invasive mole, and choriocarcinoma. Extravillous cytotrophoblast (EVT) cells were positive for both proteoglycans in all pathologies and in first-trimester placentas, although not in term placentas. Biglycan was immunolocalized in the ECM of all healthy and pathological placentas, whereas decorin was observed only in term placenta ECM. CONCLUSIONS: The expression of both proteoglycans was cell-specific and gestation time-dependent in healthy placentas, and was associated with invasive EVT cells in pathological placentas. In view of the biological properties of these molecules, it is possible that the biglycan pattern found here is intrinsically implicated in the invasive activity of EVT cells in both healthy and disordered placentas.

Angiogenic factors and uterine Doppler velocimetry in early- and late-onset preeclampsia.

OBJECTIVE: To assess correlations between maternal serum levels of pro- and anti-angiogenic factors with uterine perfusion in women with early- compared with late-onset preeclampsia, and in healthy pregnant women. DESIGN: Case-control study. SETTING: Antenatal care clinic located within a hospital (São Bernardo do Campo, Brazil). POPULATION: We enrolled 54 preeclamptic and 54 healthy control women who were coming for routine ultrasound at 28-36 weeks' gestation. METHODS: All participants had uterine artery and umbilical Doppler studies and a blood sample to assess maternal serum levels of soluble fms-like tyrosine kinase-1, soluble endoglin, adiponectin and plasminogen activator inhibitor-1. All angiogenic factors were measured using enzyme-linked immunosorbent assay. MAIN OUTCOME MEASURES: Levels of pro- and anti-angiogenic factors in maternal serum, and uterine artery Doppler findings. RESULTS: Concentrations of soluble fms-like tyrosine kinase-1 and soluble endoglin were significantly higher in preeclamptic than control women (p < 0.0001 and p < 0.0001, respectively), especially in those with early-onset (<34 weeks) preeclampsia. These two anti-angiogenic mediators were significantly correlated with increased uterine artery Doppler in the preeclamptic women. Plasminogen activator inhibitor-1 levels were significantly higher in preeclampsia (p = 0.03) but unrelated to uterine artery resistance. Adiponectin levels were similar in cases and controls, independent of body mass index and unrelated to uterine artery resistance. CONCLUSION: Preeclamptic patients have increased soluble fms-like tyrosine kinase-1 and soluble endoglin serum levels and this increase is directly correlated with uterine artery resistance, especially in those with early-onset preeclampsia.

Serum levels and gestational curve of adiponectin and leptin during adolescent pregnancy.

OBJECTIVE: This study aimed to develop a curve of weekly serum levels of adiponectin and leptin among pregnant adolescents. In addition, pregestational body mass index and weight gain were assessed and correlated with the serum concentration of these molecules. METHODS: This was a prospective cohort study, including only pregnant adolescents with eutrophic pre-gestational body mass index who were weekly followed during the evolution of gestation. The serum concentrations of adipokines were determined using commercial ELISA kits and were correlated to pre-gestational body mass index and pregnancy weight gain. A total of 157 pregnant women participated in this study. RESULTS: Adiponectin levels showed a significant decrease among the trimesters (p=0.0004). However, we did not observe significant differences among its levels when compared weekly, neither of which was between adiponectin concentration and pre-gestational body mass index or weight gain (p=0.36 and p=0.10, respectively). In contrast, we detected a significant increase in weekly serum leptin levels (p<0.0001), positively correlated to both pre-gestational body mass index and weight gain (p=0.003 and p=0.0007, respectively). CONCLUSION: These adipokines present a different profile throughout adolescent pregnancy.

Adiponectin and leptin serum levels in normal adolescent pregnancies.

INTRODUCTION: Adiponectin and leptin play a critical role in pregnancy development, the blood levels of these adipokines have been extensively investigated in healthy adult women, however there are no similar studies in adolescents. The aim of this study was to evaluate adiponectin and leptin serum levels in adolescents during pregnancy. METHODS: This prospective cohort study recruited 105 healthy, normal-weight adolescents, within the ages from 13 to 19 years old. Leptin and adiponectin serum levels of the 43 pregnant participants were assessed at 10-14, 24-28 and 30-34 weeks and their concentrations were compared with those of the 62 nonpregnant adolescents. Commercial ELISA kits were used for all assessments. RESULTS: There were no clinical and sociodemographic differences between the pregnant and nonpregnant adolescents. Adiponectin serum levels were significantly lower in the pregnant compared to the nonpregnant adolescents 3600 ± 1730 ng/ml versus 4144 ± 1583 ng/ml, respectively. (p < .0001). Moreover, adiponectin concentration decreased significantly with pregnancy progress: being 4295 (± 2003) ng/ml at the first trimester; 3419 (±1803) ng/ml at the 2nd; and 3112 (±1442) ng/ml at the 3rd trimesters, respectively (p = .004). Overall leptin serum concentrations were significantly higher in pregnant than in nonpregnant adolescents (p < .0001). During pregnancy, leptin concentration increased significantly between the first and third trimesters: 35,688 (±33,637) pg/ml versus 49,388 (±33,186) pg/ml, respectively, (p < .0001). CONCLUSIONS: The profile of adiponectin and leptin serum levels in adolescent are similar to that observed in adult women. In both cases, there are significant changes with gestational age during healthy pregnancy.Key MessageAdiponectin and leptin serum levels in healthy teenager changes with pregnancy.

Evaluation of the Blood Level of Adiponectin in Pregnant Adolescents.

OBJECTIVE: To evaluate serum levels of adiponectin in pregnant adolescents between 30 and 36 weeks of gestation. METHOD: A prospective cross-sectional study enrolled 67 normal pregnant women between 30 and 36 weeks of gestation and eutrophic (body mass index [BMI]: 18.5-25 kg/m2), of which 36 were adolescents (< 20 years old) and 31 adults (≥ 20 years old). Serum adiponectin levels were determined by enzyme-linked immunosorbent assay (ELISA). The t-student or Mann-Whitney tests were used for intergroup comparison. RESULTS: Pregnant adolescents showed significantly higher serum adiponectin concentrations compared with pregnant adults (p = 0.04). No differences were observed in adiponectin levels in younger pregnant adolescents (< 16 years old) compared with older pregnant adolescents (≥ 16 years old). Adiponectin values were divided into 3 subgroups: < 3,000 ng/mL, between 3,000 and 5,000 ng/mL, and > 5,000 ng/mL. Birthweight was significantly higher in women > 5,000 ng/mL when compared with < 3,000 ng/mL in the adolescent group. No association between pregestational adiponectin levels and BMI, gestational weight gain, and gestational age was observed; however, there was a positive relation with birthweight (p = 0.0239). CONCLUSION: Serum adiponectin values in pregnant adolescents between 30 and 36 weeks of gestation were higher compared with pregnant adults; however, no differences between younger and older pregnant adolescents were observed.

OBJETIVO: Avaliar os níveis séricos de adiponectina em gestantes adolescentes entre 30 e 36 semanas de gestação. MéTODOS: Estudo prospectivo e transversal incluindo 67 gestantes normais entre 30 a 36 semanas e eutróficas (índice de massa corporal [IMC]: 18,5­25 kg/m2), sendo 36 adolescentes (< 20 anos) e 31 adultas (≥ 20 anos). Os níveis séricos de adiponectina foram avaliados por teste imunoenzimático (ELISA, na sigla em inglês). Para a comparação entre os grupos, utilizou-se os testes t-Student ou Mann-Whitney. RESULTADOS: As gestantes adolescentes apresentaram significativamente maiores concentrações séricas de adiponectina do que as adultas (p = 0,04). Não houve diferenças nos níveis de adiponectina quando comparadas as gestantes adolescentes precoces (< 16 anos) às tardias (≥ 16 anos). Os valores de adiponectina foram subdivididos em 3 grupos: < 3.000 ng/mL, entre 3.000 e 5.000 ng/mL e > 5.000 ng/mL. O peso do recém-nascido foi significantemente maior nas mulheres com > 5.000 ng/mL, quando comparadas as com < 3.000 ng/mL no grupo das adolescentes. Não foi observada associação entre os níveis de adiponectina e o IMC pré-gestacional, ganho de peso gestacional e a idade gestacional, porém houve relação positiva com o peso do recém-nascido (p = 0,0239). CONCLUSãO: Os valores séricos de adiponectina em gestantes adolescentes entre 30 e 36 semanas de gestação foram maiores do que os das gestantes adultas; contudo, sem diferenças entre gestantes adolescentes precoces e tardias.

The effect of Gestational Diabetes Mellitus on the fetal compartment.

In indicated preterm births such a Gestational Diabetes Mellitus (GDM), little is known about the role of the amnion membranes. Investigating the role of amnion membrane inflammation in response GDM may suggest novel pathophysiologic mechanisms. We hypothesize that increased GDM inflammatory mediators may weaken the amnion membrane predisposing them to infection. Maternal and fetal serum and amnion membrane biopsies were collected from 20 GDM and 38 normoglycemic subjects (control) who underwent elective cesarean sections. Cytokines and adipokines were evaluated in serum and amnion culture supernatant samples. Amnion membrane biopsies from GDM and control subjects were studied: fresh frozen for RNA analysis for Toll-like receptor expression; cultured with LPS to test membrane permeability, and inflammation LPS + anti-TLR4 for testing mechanism. GDM was associated with higher fetal serum leptin (p = 0.004) and IL-10 (p = 0.04) compared to controls. Amnion membrane explants from GDM had higher levels of IL-6 (p = 0.019), and lower expression of Claudin-4 (p = 0.007) and increased permeability (p = 0.046) compared to controls. GDM membranes treated with LPS showed an increased expression of IL-10 (p = 0.013); IL-6 (p = 0.004) and TNF-α (p = 0.0005) but did not affect membrane permeability. LPS and anti-TLR4 antibody treatment reduced the production of TNF-α in controls (p = 0.03) and GDM (p = 0.007) compared to LPS alone. Fetal inflammatory response seems more balanced in GDM and does not impact membrane permeability function even with an infectious stimulus. Light fetal membrane inflammatory response may explain lack of preterm labor in GDM. Concluding, benign inflammation in the membranes may not be harmful for pregnancy maintenance.

The Assessment of Vitamin D Levels in Pregnant Women is not Associated to Fetal Growth Restriction: A Cross Sectional Study.

OBJECTIVE: To assess maternal serum levels of vitamin D in fetuses appropriate for gestational age (AGA), small for gestational age (SGA), and with fetal growth restriction (FGR) according to estimated fetal weight (EFW). METHODS: This cross-sectional study included 87 pregnant women between 26 and 36 weeks of gestation: 38 in the AGA group, 24 in the SGA group, and 25 in the FGR group. Maternal serum vitamin D levels were assessed using the chemiluminescence method. The Fisher exact test was used to compare the results between the groups. RESULTS: The mean ± standard deviation (SD) of maternal age (years) and body mass index (kg/m2) in the AGA, SGA, and FGR groups were 25.26 ± 8.40 / 26.57 ± 4.37; 25.04 ± 8.44 / 26.09 ± 3.94; and 25.48 ± 7.52 / 26.24 ± 4.66, respectively (p > 0.05). The maternal serum vitamin D levels (mean ± SD) of the AGA, SGA, and FGR groups were 22.47 ± 8.35 ng/mL, 24.80 ± 10.76 ng/mL, and 23.61 ± 9.98 ng/mL, respectively, but without significant differences between the groups (p = 0.672). CONCLUSION: Maternal serum vitamin D levels did not present significant differences among pregnant women with AGA, SGA, or FGR fetuses between 26 and 36 weeks of gestation according to EFW.

OBJETIVO: Avaliar o nível sérico materno de vitamina D em fetos adequados para idade gestacional (AIG), pequenos para idade gestacional (PIG) e com restrição de crescimento (RCF) de acordo com a estimativa de peso fetal (EPF). MéTODOS: Realizou-se um estudo transversal envolvendo 87 gestantes entre 26 e 36 semanas, sendo: 38 do grupo AIG, 24 do grupo PIG e 25 do grupo RCF. A dosagem sérica materna de vitamina D foi realizada pelo método de quimiluminescência. Para as comparações entre os grupos, utilizou-se o teste exato de Fisher. RESULTADOS: A média ± desvio-padrão (DP) da idade materna (anos) e do índice de massa corporal (kg/m2) nos grupos AIG, PIG e RCF foram 25,26 ± 8,40 / 26,57 ± 4,37; 25,04 ± 8,44 / 26,09 ± 3,94; e 25,48 ± 7,52 / 26,24 ± 4,66, respectivamente (p > 0,05). A concentração sérica materna de vitamina D (médias ± desvios-padrão) dos grupos AIG, PG e RCF foram 22,47 ± 8,35 ng/ml; 24,80 ± 10,76 ng/ml; e 23,61 ± 9,98 ng/ml, respectivamente, contudo, sem diferenças significativas entre os grupos (p = 0,672). CONCLUSãO: A concentração sérica materna de vitamina D não apresentou diferenças significantes entre gestantes com fetos AIG, PIG ou RCF entre 26 e 36 semanas de acordo com a EPF.

Noninvasive determination of fetal rh blood group, D antigen status by cell-free DNA analysis in maternal plasma: experience in a Brazilian population.

We evaluated the diagnostic accuracy of Rh blood group, D antigen (RHD) fetal genotyping, using real-time polymerase chain reaction in maternal blood samples, in a racially mixed population. We performed a prospective study conducted between January 2006 and December 2007, analyzing fetal RHD genotype in the plasma of 102 D- pregnant women by real-time polymerase chain reaction, targeting exons 7 and 10 of the RHD gene. Genotype results were compared with cord blood phenotype obtained after delivery or before the first intrauterine transfusion when necessary. Most of the participants (75.5%) were under 28 weeks of pregnancy, and 87.5% had at least one relative of black ancestry. By combining amplification of two exons, the accuracy of genotyping was 98%, sensitivity was 100%, and specificity was 92%. The positive likelihood ratio was 12.5, and the negative likelihood ratio was 0. The two false-positive cases were confirmed to be pseudogene RHD by real-time polymerase chain reaction. There were no differences between the patients with positive or negative Coombs test ( P = 0.479). Determination of fetal RHD status in maternal peripheral blood was highly sensitive in this racially mixed population and was not influenced by the presence of antierythrocyte antibodies.

Blood level of adipokines and nutritional status variables in adolescent pregnancy.

OBJECTIVE: To evaluate the serum levels of adiponectin and leptin and their relationship with nutritional variables during pregnancy in adolescents. METHODS: This prospective cohort study evaluated eutrophic pregnant adolescents (body mass index [BMI], 18.5-24.9 kg/m2) during the 3 gestational trimesters (first, 10-14 weeks; second, 24-28 weeks; and third, 30-34 weeks). Serum adiponectin and leptin concentrations were measured using the enzyme-linked immunosorbent assay method. The relationship of these adipokines with the pre-gestational BMI, gestational weight gain, weight at the time of sample collection, and newborn weight were evaluated. Analysis of variance and the Kruskal-Wallis test were used for statistical analysis. RESULTS: The study group comprised 62 pregnant adolescents. The serum concentration of adiponectin showed a significant difference between the first and third trimesters (P=0.003), which decreased during pregnancy, but unrelated to nutritional variables. Serum leptin levels increased throughout the pregnancy (P<0.0001) and showed a positive correlation with pre-gestational BMI, total weight gain, pregnancy weight at the time of sample collection, and newborns' weight. CONCLUSION: Serum levels of adiponectin and leptin vary inversely throughout pregnancy. This pattern in adolescents is similar to that observed in adults. Moreover, leptin concentrations increased throughout pregnancy, and they were positively correlated with all variables evaluated.

The chimera-type galectin-3 is a positive modulator of trophoblast functions with dysregulated expression in gestational diabetes mellitus.

PROBLEM: From conception, a delicate regulation of galectins, a family of carbohydrate-binding proteins, is established to ensure maternal immune tolerance in pregnancy. Though galectin-3 (gal-3), the only chimera-type galectin, is abundantly expressed at the feto-maternal interface; the physiological role of this lectin during pregnancy remains to be fully elucidated and requires further investigation. METHOD OF STUDY: In this study, we analyzed serum gal-3 levels during the course of healthy gestation. Trophoblast functions were evaluated upon gal-3 exogenous stimulation using trophoblastic cell lines (e.g. , HIPEC65, SGHPL-4, and BeWo cells). Finally, we investigated variations in peripheral gal-3 levels associated with the development of spontaneous abortion and gestational diabetes mellitus (GDM). RESULTS: Gal-3 circulating levels increased as normal pregnancy progressed. In vitro experiments showed that exogenous gal-3 positively regulated trophoblast functions inducing invasion, tube formation, and fusion. Compared with normal pregnant women, circulating gal-3 levels were significantly decreased in patients who developed GDM. CONCLUSION: Our results reveal a physiological role for gal-3 during pregnancy, promoting proper trophoblast functions associated with healthy gestation. GDM is associated with a failure to increase circulating gal-3 levels late in gestation. Thus, dysregulation of gal-3 may indicate a contribution of the chimera-type lectin to this adverse pregnancy outcome.

Fat Grafting for Treatment of Facial Burns and Burn Scars.

This article presents the authors' experience with the use of fat grafting via the Coleman technique, for the adjuvant treatment of facial burn wounds and their sequelae. It demonstrates the regenerative effects of fat injected under the wound and/or the scar as well as of fat delivered to the debrided surface of the wound and to the surface of the scar after laser treatment or microneedling.

Assessment of Polymorphism of the VDR Gene and Serum Vitamin D Values in Gestational Diabetes Mellitus.

OBJECTIVE: To evaluate the relationship between vitamin D receptor (VDR) gene polymorphism (FokI [rs10735810]) and serum vitamin D concentration in gestational diabetes mellitus (GDM). METHODS: A prospective case-control study that recruited healthy pregnant women (control group) (n = 78) and women with GDM (GDM group) (n = 79), with no other comorbidities. Peripheral blood samples were collected in the 3rd trimester of gestation, and all of the pregnant women were followed-up until the end of the pregnancy and the postpartum period. Serum vitamin D concentrations were measured by high-performance liquid chromatography (HPLC). For genomic polymorphism analysis, the genomic DNA was extracted by the dodecyltrimethylammonium bromide/cetyltrimethylammonium bromide (DTAB/CTAB) method, and genotyping was performed by the polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) technique, using the restriction enzyme FokI. The Student-t, Mann-Whitney, chi-squared, and Fischer exact tests were used for the analysis of the results. RESULTS: There was no significant difference between the pregnant women in the control and GDM groups regarding serum vitamin D levels (17.60 ± 8.89 ng/mL versus 23.60 ± 10.68 ng/mL; p = 0.1). Also, no significant difference was detected between the FokI genotypic frequency when the 2 groups were compared with each other (p = 0.41). CONCLUSION: There was no association between the FokI polymorphism and the development of GDM, nor was there any change in serum vitamin D levels in patients with GDM.

OBJETIVO: Avaliar a relação entre o polimorfismo do gene receptor da vitamina D (VDR) (FokI [rs10735810]) e a concentração sérica de vitamina D no diabetes mellitus gestacional (DMG). MéTODOS: Estudo prospectivo tipo caso-controle que recrutou gestantes saudáveis (grupo controle) (n = 78) e com DMG (grupo DMG) (n = 79), sem outras comorbidades. Foram coletadas amostras de sangue periférico no 3° trimestre da gestação, e todas as gestantes foram acompanhadas até o final da gravidez e no pós-parto. As concentrações séricas de vitamina D foram mensuradas por cromotografia líquida de alta eficiência (CLAE). Para análise do polimorfismo genético, o DNA genômico foi extraído pelo método de brometo de dodeciltrimetilamônio/brometo de cetiltrimetilamônio (DTAB/CTAB), e as genotipagens foram realizadas por técnica de reação de cadeia de polimerase ­ polimorfismo do comprimento do fragmento de restrição (PCR-RFLP, na sigla em inglês), sendo empregada a enzima de restrição FokI. Foram utilizados os testes t-Student, Mann-Whitney, qui-quadrado e exato de Fischer para a análise dos resultados. RESULTADOS: Não houve diferença significativa entre as gestantes dos grupos controle e DMG quanto aos níveis séricos de vitamina D (17,60 ± 8,89 ng/mL versus 23,60 ± 10,68 ng/mL; p = 0,1). Também não foi detectada diferença significativa entre a frequência genotípica de FokI, quando comparados os 2 grupos entre si (p = 0,41). CONCLUSãO: Não foi identificada associação do polimorfismo FokI com o desenvolvimento de DMG, bem como não foi observada alteração nos níveis séricos de vitamina D em pacientes com DMG.