Problem The immune cellular composition of amniotic fluid is poorly understood. Herein, we determined the immunophenotype of amniotic fluid: 1) immune cells during the second and third trimester; 2) T cells and innate lymphoid cells (ILCs); and 3) immune cells during intra-amniotic infection/inflammation. Method of Study Amniotic fluid samples (n=57) were collected from women from 15-40 weeks of gestation without intra-amniotic infection/inflammation. Samples from women with intra-amniotic infection/inflammation were also included (n=9). Peripheral blood mononuclear cells from healthy adults were used as controls (n=3). Immunophenotyping was performed using flow cytometry. Results In the absence of intra-amniotic infection/inflammation, the amniotic fluid contained several immune cell populations from 15-40 weeks. Among these immune cells: 1) T cells and ILCs were greater than B cells and NK cells between 15 to 30 weeks; 2) T cells were most abundant between 15 to 30 weeks; 3) ILCs were most abundant between 15 to 20 weeks; 4) B cells were scarce between 15 to 20 weeks; yet, they increased and were constant after 20 weeks; 5) NK cells were greater between 15 to 30 weeks than at term; 6) ILCs expressed high levels of RORγt, CD161, and CD103 (i.e. Group 3 ILCs); 7) T cells expressed high levels of RORγt; 8) neutrophils increased as gestation progressed; and 9) monocytes/macrophages emerged after 20 weeks and remained constant until term. All of the amniotic fluid immune cells, except ILCs, were increased in the presence of intra-amniotic infection/inflammation. Conclusions The amniotic fluid harbors a diverse immune cellular composition during normal and complicated pregnancies.

Problem: Preterm birth is commonly preceded by preterm labor, a syndrome that is causally linked to both intra-amniotic infection and intra-amniotic inflammation.However, the stereotypical cellular immune responses in these two clinical conditions are poorly understood.Method of study: Amniotic fluid samples (n = 26) were collected from women diagnosed with preterm labor and intra-amniotic infection (amniotic fluid IL-6 concentrations ≥2.6 ng/mL and culturable microorganisms, n = 10) or intra-amniotic inflammation (amniotic fluid IL-6 concentrations ≥2.6 ng/mL without culturable microorganisms, n = 16). Flow cytometry was performed to evaluate the phenotype and number of amniotic fluid leukocytes. Amniotic fluid concentrations of classical proinflammatory cytokines, type 1 and type 2 cytokines, and T-cell chemokines were determined using immunoassays.Results: Women with spontaneous preterm labor and intra-amniotic infection had (a) a greater number of total leukocytes, including neutrophils and monocytes/macrophages, in amniotic fluid; (b) a higher number of total T cells and CD4 + T cells, but not CD8 + T cells or B cells, in amniotic fluid; and (c) increased amniotic fluid concentrations of IL-6, IL-1β, and IL-10, compared to those with intra-amniotic inflammation. However, no differences in amniotic fluid concentrations of T-cell cytokines and chemokines were observed between these two clinical conditions. K E Y W O R D S chorioamnionitis, fetal inflammatory response syndrome, funisitis, microbial invasion of the amniotic cavity, placental inflammation, pregnancy | 3 of 15 GOMEZ-LOPEZ et al. | Placental histopathological examinationPlacentas were examined histologically by perinatal pathologists blinded to clinical diagnoses and obstetrical outcomes according to standardized Perinatology Research Branch protocols. 75,76 Briefly, three to nine sections of the placenta were examined, and at least one full-thickness section was taken from the center of the placenta; others were taken randomly from the placental disk. Acute inflammatory lesions of the placenta (maternal inflammatory response and fetal inflammatory response) were diagnosed according to established criteria, including staging and grading. 75,77 The proportions of patients whose placentas presented acute maternal and/or fetal inflammatory responses are displayed in Table 1. | Amniotic fluid sample collectionAmniotic fluid samples were obtained by transabdominal amniocentesis under antiseptic conditions and monitored by ultrasound in order to detect intra-amniotic inflammation and/or infection in patients with preterm labor. Samples of amniotic fluid were transported to the laboratory in a sterile, capped syringe and immunophenotyping was performed immediately. The rest of the sample was centrifuged at 1300 g for 10 minutes at 4°C, and the supernatant was stored at −80°C until use. Also, an aliquot of amniotic fluid was transported to the clinical laboratory for culture of aerobic/anaerobic bacteria and genital mycoplasmas. The clinical tests also inc...

Fetal dextrocardia is a type of cardiac malposition where the major axis from base to apex points to the right side. This condition is usually associated with a wide spectrum of complex cardiac defects. As a result, dextrocardia is conceptually difficult to understand and diagnose on prenatal ultrasound. The advantage of four-dimensional sonography with spatiotemporal image correlation (STIC) is that this modality can facilitate fetal cardiac examination. A novel method known as fetal intelligent navigation echocardiography (FINE) allows automatic generation of nine standard fetal echocardiography views in normal hearts by applying intelligent navigation technology to STIC volume datasets. In fetuses with congenital heart disease, FINE is also able to demonstrate abnormal cardiac anatomy and relationships when there is normal cardiac axis and position. However, this technology has never been applied to cases of cardiac malposition. We report herein for the first time, a case of fetal dextrocardia and situs solitus with complex congenital heart disease in which the FINE method was invaluable in diagnosing multiple abnormalities and defining complex anatomic relationships. We also review the literature on prenatal sonographic diagnosis of dextrocardia (with an emphasis on situs solitus), as well as tricuspid atresia with its associated cardiac features.