Isolation and Immunophenotyping of Leukocytes from the Human Maternal-Fetal Interface.

During human pregnancy, leukocytes that infiltrate the maternal-fetal interface play a major role in establishing a delicate balance between immune tolerance and functional response and setting the inflammatory process that leads to labor. Here we describe two methods for isolating immune cells from the chorioamniotic membranes (decidua parietalis) and placental blood (decidua basalis) that combine gentle enzymatic digestion, magnetic cell sorting, and density gradient. Isolated leukocytes can be immunophenotypified by flow cytometry, and both isolation methods are compatible with downstream cellular and molecular applications, such as cell culture, transcriptome, and proteome analyses.

The Intuitive Eating Scale-2 Adapted for Mexican Pregnant Women: Psychometric Properties and Influence of Sociodemographic Variables.

A weight-inclusive approach to health involves the promotion of intuitive eating, i.e., the individual's ability to be aware of their physiological hunger and satiety cues to determine when and how much to eat, while paying attention to how certain foods affect their body.

A Mexican Spanish Adaptation of the Dutch Eating Behavior Questionnaire: Psychometric Properties and Influence of Sociodemographic Variables in Pregnant Women.

Eating behaviors are complex phenomena, entangling physiological signals of hunger and satiety, food choices, emotional states, and social factors and expectations, as well as food availability and sensory appearance. Evaluating eating behaviors is challenging and must cover different motives. One instrument for such evaluation is the Dutch Eating Behavior Questionnaire (DEBQ), composed of three subscales for exploring emotional eating, external eating, and restrained eating.

Infant Feeding Practices That Substitute Exclusive Breastfeeding in a Semi-Rural Mexican Community: Types, Moments, and Associated Factors.

International organizations recommend mothers practice exclusive breastfeeding (EBF) during the first six months of their infant's life and introduce complementary feeding (CF) thereafter while continuing breastfeeding. However, the earlier introduction of liquids and foods is common worldwide and may have negative effects on breastfeeding practice, nutrition, and health. In this formative cross-sectional study, we interviewed 143 mothers from semi-rural communities in Tabasco, Mexico, whose infants were 4-6 months old.

Exclusive Breastfeeding and Factors Influencing Its Abandonment During the 1st Month Postpartum Among Women From Semi-rural Communities in Southeast Mexico.

Introduction: In this study we describe breastfeeding practices among women from semi-rural communities in southeast Mexico, and explore which factors, modifiable or not, are associated with such practices.

Materials And Methods: This was a formative cross-sectional study that included 143 mothers with infants 4-6 months old, from semi-rural communities in Tabasco, Mexico. We collected data on two categories of factors: (1) women's sociodemographic characteristics, and (2) maternal / infant factors.

Prolactin Protects the Structural Integrity of Human Fetal Membranes by Downregulating Inflammation-induced Secretion of Matrix Metalloproteinases.

Prolactin (PRL) is a pleiotropic hormone with a key role in pregnancy. In fetal membranes, PRL can regulate the secretion of pro-inflammatory factors, which induces the activation of matrix metalloproteinases (MMPs). The increase and activation of MMPs deregulate the turnover of the extracellular matrix in the fetal membranes, altering its structure and function, causing premature rupture of the membranes and preterm labor.

Maternal adiposity is associated with inflammatory gene expression in leukocytes at term human pregnancy: A pilot study.

Background: Human labor is associated with an inflammatory process that takes place at the maternal-fetal interface, where leukocytes infiltrate and contribute to the local production of effector molecules such as cytokines, chemokines, MMPs, etc. This process may be altered by a low-grade chronic inflammation, characteristic of obesity, resulting in adverse pregnancy outcomes. In this cross-sectional pilot study, we analyzed the relationship between maternal adiposity and inflammation-related gene expression in leukocytes from six healthy women with term pregnancies without labor.

Erythropoiesis and Red Cell Indices Undergo Adjustments during Pregnancy in Response to Maternal Body Size but not Inflammation.

During human pregnancy, iron requirements gradually increase, leading to higher amounts of erythropoietin (EPO) and reticulocytes, and changes in erythrocyte size and density. Women with pregestational obesity experience "obesity hypoferremia" during pregnancy, which alters iron homeostasis. In this study we aimed to describe the relationship between EPO and iron nutrition status during nonanemic pregnancy, and to explore whether obesity and inflammation influence erythropoiesis and red cell indices.

Innate Immune Cells and Toll-like Receptor-Dependent Responses at the Maternal-Fetal Interface.

During pregnancy, the placenta, the mother and the fetus exploit several mechanisms in order to avoid fetal rejection and to maintain an immunotolerant environment throughout nine months. During this time, immune cells from the fetal and maternal compartments interact to provide an adequate defense in case of an infection and to promote a tolerogenic milieu for the fetus to develop peacefully. Trophoblasts and decidual cells, together with resident natural killer cells, dendritic cells, Hofbauer cells and other macrophages, among other cell types, contribute to the modulation of the uterine environment to sustain a successful pregnancy.

Obesity Is Associated with Changes in Iron Nutrition Status and Its Homeostatic Regulation in Pregnancy.

The influence of obesity on maternal iron homeostasis and nutrition status during pregnancy remains only partially clarified. Our study objectives were (1) to describe how obesity influences broad iron nutrition spectrum biomarkers such as available or circulating iron (serum transferrin receptor (sTfr) and serum iron), iron reserves (ferritin), and functional iron (hemoglobin); and (2) to depict the regulating role of hepcidin. The above was carried out while considering influential factors such as initial iron nutrition status, iron intake, and the presence of inflammation.

A trypsin-based method for isolating leukocytes from human choriodecidua suitable for immunophenotyping and transcriptome studies.

Leukocytes found at the human maternal-fetal interface participate in the inflammatory process associated with both preterm and term labor; therein, effective methods for their isolation that allow further phenotypic and functional analyses are necessary. Leukocyte isolation is usually carried out through scraping or enzyme digestion of the choriodecidua, however both methods usually limit the use of downstream immunophenotyping or transcriptomic techniques. Here we describe an isolation method based on gentle trypsin digestion that yields a leukocyte-enriched cell mixture with high lymphocyte viability, although less viable myeloid cells.

Choriodecidual leukocytes display a unique gene expression signature in spontaneous labor at term.

Prior to and during the process of human labor, maternal circulating leukocytes infiltrate the maternal-fetal interface (choriodecidua) and become activated resembling choriodecidual leukocytes. Since, there is no evidence comparing maternal circulating and choriodecidual leukocytes, herein, we characterized their transcriptome and explored the biological processes enriched in choriodecidual leukocytes. From women undergoing spontaneous term labor we isolated circulating and choriodecidual leukocytes, performed microarray analysis (n = 5) and qRT-PCR validation (n = 9) and interaction network analysis with up-regulated genes.

Selective immuno-modulatory effect of prolactin upon pro-inflammatory response in human fetal membranes.

During pregnancy, prolactin (PRL) is a neuro-immuno-cytokine that contributes actively to the crosstalk between the immune and endocrine systems and, thus, to the creation of an immune-privileged milieu. This work aims to analyze the capacity of PRL to modulate the synthesis and secretion of pro-inflammatory markers associated with labor. Studies were conducted using human fetal membranes at term mounted in a model of two independent chambers.

Decidualization Mediated by Steroid Hormones Modulates the Innate Immunity in Response to Group B Streptococcal Infection in vitro.

Background: Decidual cells play a role in the modulation of the innate immune response to protect pregnancy against infection. Steroid hormones regulate the innate immune response in different tissues, and they are involved in several biological processes like decidualization. The aim of this study was to assess if steroid hormones modulate the innate immunity in endometrial stromal cells (ESCs) and decidual stromal cells (DSCs) in response to group B streptococcus (GBS) infection in vitro.

The potential role of prolactin as a modulator of the secretion of proinflammatory mediators in chorioamniotic membranes in term human gestation.

Objective: To test the effect of prolactin (PRL) on expression of proinflammatory cytokines and matrix metallopeptidase 9 (MMP-9) in vitro.

Study Design: Tissue explants were incubated from 4 to 48 hours alone or in the presence of 500 ng/mL PRL, and mRNA expression in tissues and secretion of interleukin (IL)-1β, tumor necrosis factor alpha (TNF-α), MMP-2, and MMP-9 was quantified.

Results: Fetal membranes secreted IL-1β, TNF-α, and MMP-9 in culture with consistent low concentration during the first 24 hours and then increased progressively.

Tissue-specific IL-10 secretion profile from term human fetal membranes stimulated with pathogenic microorganisms associated with preterm labor in a two-compartment tissue culture system.

Objective: Interleukin (IL)-10 is a cytokine with anti-inflammatory properties that plays pivotal roles in immune recognition and maintenance of pregnancy, limiting the harmful effects of pro-inflammatory modulators. The aim of this work was to characterize the contribution of amnion and choriodecidua regions of the human fetal membranes in the production of IL-10 after selective stimulation with Candida albicans, Gardnerella vaginalis and Streptococcus agalactiae.

Methods: Pre-labor human fetal membranes were cultured in a two-compartment tissue culture system and stimulated with 1 × 10(6) CFU/ml of each pathogen added to either the amniotic or choriodecidual region or both.

Progesterone elicits an inhibitory effect upon LPS-induced innate immune response in pre-labor human amniotic epithelium.

Problem: Infection of human fetal membranes elicits secretion of pro-inflammatory modulators through its innate immune capacities. We investigated the effect of lipopolysacharide (LPS) and progesterone (P4) upon expression of TLR-4/MyD88, TNFα, IL-6, IL-8, IL-10, and HBD2 on the human amniotic epithelium.

Method Of Study: Explants of the human amniotic epithelium were pre-treated with 0.

Housekeeping gene expression stability in reproductive tissues after mitogen stimulation.

Background: Intrauterine infection during pregnancy can trigger a local inflammatory response leading to several complications, such as preterm labor. Many studies have used in vitro and in vivo models employing mitogens to induce the expression of the characteristic proinflammatory mediators triggered by infection. However, relative expression assays depend on the stability of housekeeping gene expression, which can vary depending on certain stimuli.

Evidence for a role for the adaptive immune response in human term parturition.

Problem: Spontaneous labor at term involves leukocyte recruitment and infiltration into the choriodecidua; yet, characterization of these leukocytes and their immunological mediators is incomplete. The purpose of this study was to characterize the immunophenotype of choriodecidual leukocytes as well as the expression of inflammatory mediators in human spontaneous parturition at term.

Method Of Study: Choriodecidual leukocytes were analyzed by FACS, immunohistochemistry, and RT-PCR in three different groups: (i) preterm gestation delivered for medical indications without labor; (ii) term pregnancy without labor; and (iii) term pregnancy after spontaneous labor.

Tissue-specific human beta-defensins (HBD)-1, HBD-2 and HBD-3 secretion profile from human amniochorionic membranes stimulated with Candida albicans in a two-compartment tissue culture system.

Background: During intrauterine infection, amniochorionic membranes represent a mechanical and immunological barrier against dissemination of infection. Human beta defensins (HBD)-1, HBD-2, and HBD-3 are key elements of innate immunity that represent the first line of defense against different pathogen microorganisms associated with preterm labor. The aim of this work was to characterize the individual contribution of the amnion (AMN) and choriodecidua (CHD) regions to the secretion of HBD-1, HBD-2 and HBD-3, after stimulation with Candida albicans.

Weight-based nutritional diagnosis of Mexican children and adolescents with neuromotor disabilities.

Background: Nutrition related problems are increasing worldwide but they have scarcely been evaluated in people with neuromotor disabilities, particularly in developing countries. In this study our aim was to describe the weight-based nutritional diagnoses of children and adolescents with neuromotor disabilities who attended a private rehabilitation center in Mexico City.

Methods: Data from the first visit's clinical records of 410 patients who attended the Nutrition department at the Teleton Center for Children Rehabilitation, between 1999 and 2008, were analyzed.

[Expression of cell adhesion molecules in the maternal fetal period of human gestation].

Background: The human labor is an inflammatory process invading leukocytes modulated by gestational tissues. The local increase of cell adhesion molecules (CAMs) promotes the permanence of these leukocytes in the coriodecidua. Gestational tissues express ICAM-1, while circulating leukocytes expressing its ligand Mac-1.

Association between adiposity and inflammatory markers in maternal and fetal blood in a group of Mexican pregnant women.

In the present pilot study, we evaluated the effect of maternal adiposity on the plasma concentration of adipocytokines in pregnant women and their newborns. Twenty patients with term gestations without labour were initially selected by pregestational BMI and then classified into two study groups (n 10 each), according to their median value of adiposity (total body fat). Concentrations of TNF-α, IL-1β, IL-6, leptin and adiponectin in plasma of maternal peripheral blood and fetal cord blood were measured and correlated to maternal adiposity.

Placental blood leukocytes are functional and phenotypically different than peripheral leukocytes during human labor.

Rupture of the fetal membranes during human labor is associated with an inflammatory process localized to the maternal-fetal interface. There is evidence that specific leukocytes subsets are attracted to the choriodecidua, and that after homing they condition a local inflammatory microenvironment, possibly being directly involved in rupture of the membranes. In this study our aim was to compare the phenotypes and function of leukocytes located in the placental intervillous blood with peripheral leukocytes obtained before or after labor, including expression of modulators of inflammation in these cells.

[Three-dimensional analysis of human chorioamniotic membranes structure in term pregnancy].

Background: Physical and structural chorioamniotic membranes integrity is due to a precise process of synthesis and degradation of collagen; surrounding collagenolitic activity raises during labor, what leads to a structural loss and mechanical resistance weakening, the main cause of its rupture under physiological and pathological conditions. Understanding of its three-dimensional structure is essential to characterize normal and pathological labor.

Objective: To analyze three-dimensional structure of human chorioamniotic membranes at gestational term.