Optimizing perinatal wellbeing in pregnancy with obesity: a clinical trial with a multi-component nutrition intervention for prevention of gestational diabetes and infant growth and neurodevelopment impairment.

Pregnancy complicated by obesity represents an increased risk of unfavorable perinatal outcomes such as gestational diabetes mellitus (GDM), hypertensive disorders in pregnancy, preterm birth, and impaired fetal growth, among others. Obesity is associated with deficiencies of micronutrients, and pregnant women with obesity may have higher needs. The intrauterine environment in pregnancies complicated with obesity is characterized by inflammation and oxidative stress, where maternal nutrition and metabolic status have significant influence and are critical in maternal health and in fetal programming of health in the offspring later in life. Comprehensive lifestyle interventions, including intensive nutrition care, are associated with a lower risk of adverse perinatal outcomes. Routine supplementation during pregnancy includes folic acid and iron; other nutrient supplementation is recommended for high-risk women or women in low-middle income countries. This study is an open label randomized clinical trial of parallel groups (UMIN Clinical Trials Registry: UMIN000052753, https://center6.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000060194) to evaluate the effect of an intensive nutrition therapy and nutrient supplementation intervention (folic acid, iron, vitamin D, omega 3 fatty acids, myo-inositol and micronutrients) in pregnant women with obesity on the prevention of GDM, other perinatal outcomes, maternal and newborn nutritional status, and infant growth, adiposity, and neurodevelopment compared to usual care. Given the absence of established nutritional guidelines for managing obesity during pregnancy, there is a pressing need to develop and implement new nutritional programs to enhance perinatal outcomes.

Vitamin D Deficiency, Excessive Gestational Weight Gain, and Oxidative Stress Predict Small for Gestational Age Newborns Using an Artificial Neural Network Model.

(1) Background: Size at birth is an important early determinant of health later in life. The prevalence of small for gestational age (SGA) newborns is high worldwide and may be associated with maternal nutritional and metabolic factors. Thus, estimation of fetal growth is warranted. (2) Methods: In this work, we developed an artificial neural network (ANN) model based on first-trimester maternal body fat composition, biochemical and oxidative stress biomarkers, and gestational weight gain (GWG) to predict an SGA newborn in pregnancies with or without obesity. A sensibility analysis to classify maternal features was conducted, and a simulator based on the ANN algorithm was constructed to predict the SGA outcome. Several predictions were performed by varying the most critical maternal features attained by the model to obtain different scenarios leading to SGA. (3) Results: The ANN model showed good performance between the actual and simulated data (R2 = 0.938) and an AUROC of 0.8 on an independent dataset. The top-five maternal predictors in the first trimester were protein and lipid oxidation biomarkers (carbonylated proteins and malondialdehyde), GWG, vitamin D, and total antioxidant capacity. Finally, excessive GWG and redox imbalance predicted SGA newborns in the implemented simulator. Significantly, vitamin D deficiency also predicted simulated SGA independently of GWG or redox status. (4) Conclusions: The study provided a computational model for the early prediction of SGA, in addition to a promising simulator that facilitates hypothesis-driven constructions, to be further validated as an application.

Gestational Weight Gain Influences the Adipokine-Oxidative Stress Association during Pregnancy.

INTRODUCTION AND OBJECTIVE: The weight gained during pregnancy could determine the immediate and future health of the mother-child dyad. Excessive gestational weight gain (EGWG) due to abnormal adipose tissue (AT) accumulation is strongly associated with adverse perinatal outcomes as gestational diabetes, macrosomia, obesity, and hypertension further in life. Dysregulation of adipokine, AT dysfunction, and an imbalance in the prooxidant-antioxidant systems are critical features in altered AT accumulation. This study was aimed to investigate the association between adipokines and oxidative stress markers in pregnant women and the influence of the GWG on this association. METHODS: Maternal blood samples were obtained in the third trimester of pregnancy (n = 74) and serum adipokines (adiponectin, leptin, and resistin), oxidative damage markers: 8-oxo-2'-deoxyguanosine (8-oxodG), lipohydroperoxides (LOOH), malondialdehyde (MDA), and carbonylated proteins (CP), and glucose a metabolic marker were measured. RESULTS: Women with EGWG had low adiponectin levels than women with adequate weight gain (AWG) or insufficient weight gain (IWG). Multiple linear regression models revealed a positive association between adiponectin and 8-oxodG in women with AWG (B = 1.09, 95% CI: 164-222, p = 0.027) and IWG (B = 0.860, 95% CI: 0.199-1.52, p = 0.013) but not in women with EGWG. In women with EGWG, leptin was positively associated with LOOH (p = 0.018), MDA (p = 0.005), and CP (p = 0.010) oxidative markers. CONCLUSION: Our findings suggest that concurrent mechanisms regulate adipokine production and oxidative stress in pregnant women and that this regulation is influenced by GWG, probably due to an excessive AT accumulation.

Prevalence of Depression, Anxiety, and Perceived Stress in Postpartum Mexican Women during the COVID-19 Lockdown.

The COVID-19 lockdown represents a new challenge for mental health researchers and clinical practitioners. This cross-sectional study aimed to investigate the prevalence of depression, anxiety, and perceived stress in postpartum Mexican women. The study included 293, 4-12-week postpartum women over the age of 18. The Edinburgh Postpartum Depression Scale (EPDS), Trait-State Trait Anxiety Inventory (T-STAI), and Ten Perceived Stress Scale (PSS-10), which are all questionnaires validated for the Mexican population, were applied using a web-based online survey. Prevalence and 95% confidence intervals (CIs) were calculated. The mean ± standard deviation (SD) of the maternal age was 29.9 ± 6.3 years; the EPDS score: 11 ± 6, T-STAI score: 41.7 ± 12.3, and PSS-10 score: 17.1 ± 7. The prevalence (95% CI) of the postpartum depression symptoms was 39.2% (34-45%), trait anxiety symptoms were found among 46.1% (32-43%) of the participants, and moderate and high perceived stress were in 58% (52-64) and 10.9% (7.8-15) of the participants, respectively. The prevalence of depressive symptoms, generalized anxiety, and perceived stress was higher among postpartum Mexican women during the COVID-19 outbreak than before the lockdown. Our findings highlight the importance of monitoring perinatal mental health during pandemics and the need to design effective psychologic interventions for these patients.

Isolation of Viable Adipocytes and Stromal Vascular Fraction from Human Visceral Adipose Tissue Suitable for RNA Analysis and Macrophage Phenotyping.

Visceral adipose tissue (VAT) is an active metabolic organ composed mainly of mature adipocytes and stromal vascular fraction (SVF) cells, which release different bioactive molecules that control metabolic, hormonal, and immune processes; currently, it is unclear how these processes are regulated within the adipose tissue. Therefore, the development of methods evaluating the contribution of each cell population to the pathophysiology of adipose tissue is crucial. This protocol describes the isolation steps and provides the necessary troubleshooting guidelines for efficient isolation of viable mature adipocytes and SVF from human VAT biopsies in a single process, using a collagenase enzymatic digestion technique. Moreover, the protocol is also optimized to identify macrophage subsets and perform mature adipocyte RNA isolation for gene expression studies, which allows performing studies dissecting the interaction between these cell populations. Briefly, VAT biopsies are washed, minced mechanically, and digested to generate a single-cell suspension. After centrifugation, mature adipocytes are isolated by flotation from the SVF pellet. The RNA extraction protocol ensures a high yield of total RNA (including miRNAs) from adipocytes for downstream expression assays. Simultaneously, SVF cells are used to characterize macrophage subsets (pro- and anti-inflammatory phenotype) through flow cytometry analysis.

Vitamin D Deficiency in Mexican Pregnant Women: Is Supplementation with ≤400 IU/day Enough?

Controversy remains surrounding vitamin D routine supplementation in healthy pregnancy, and the doses are unclear. The aim of this study was to describe maternal vitamin D status throughout pregnancy in a group of Mexican women and evaluate the effect of frequently prescribed doses of vitamin D3 on longitudinal 25-OH-D concentrations, adjusting for obesity, season, and other factors. We conducted a cohort study (Instituto Nacional de Perinatología-INPer) (2017-2020)) of healthy pregnant women without complications. Pregestational overweight/obesity (body mass index ≥ 25), vitamin D3 supplementation (prescribed by physician; 0-250, 250-400, and >400 IU/day), and serum 25-OH-D concentrations (ELISA) were evaluated in each trimester of pregnancy. Vitamin D deficiency or insufficiency was computed (<20 and <30 ng/mL, respectively). We studied 141 adult women; 58.5% had pregestational obesity or overweight. In the first trimester, 45.8% of the women were supplemented with vitamin D3; 51.4% had vitamin D insufficiency and 37.3%, deficiency. In the third trimester, 75.4% of the women were supplemented, and 20% of them still had deficiency. The final general mixed linear model showed that 25-OH-D significantly increased throughout pregnancy (p < 0.001); the highest increase was observed in the third trimester in women with doses >400 IU/day of vitamin D3 (+4 ng/mL, 95% CI: 1.72-8.11 ng/mL). In winter/autumn, 25-OH-D concentrations were also lower (p ≤ 0.05). In this group of pregnant Mexican women, the prevalence of vitamin D deficiency and insufficiency was high. A higher increase in 25-OH-D concentrations during pregnancy was observed when the women were supplemented with >400 IU/day. Common supplementation doses of 250-400 IU/day were insufficient for achieving an adequate maternal vitamin D status.

Macrophage Populations in Visceral Adipose Tissue from Pregnant Women: Potential Role of Obesity in Maternal Inflammation.

Obesity is associated with inflammatory changes and accumulation and phenotype polarization of adipose tissue macrophages (ATMs). Obese pregnant women have alterations in adipose tissue composition, but a detailed description of macrophage population is not available. In this study, we characterized macrophage populations in visceral adipose tissue (VAT) from pregnant women with normal, overweight, and obese pregestational weight. Immunophenotyping of macrophages from VAT biopsies was performed by flow cytometry using CD45 and CD14 as markers of hematopoietic and monocyte linage, respectively, while HLA-DR, CD11c, CD163, and CD206 were used as pro- and anti-inflammatory markers. Adipocyte number and size were evaluated by light microscopy. The results show that pregnant women that were overweight and obese during the pregestational period had adipocyte hypertrophy. Two different macrophage populations in VAT were identified: recruited macrophages (CD45⁺CD14⁺), and a novel population lacking CD45, which was considered to be a resident macrophages subset (CD45−CD14⁺). The number of resident HLA−DRlow/− macrophages showed a negative correlation with body mass index (BMI). Both resident and recruited macrophages from obese women expressed higher CD206 levels. CD11c expression was higher in resident HLA-DR⁺ macrophages from obese women. A strong correlation between CD206 and CD11c markers and BMI was observed. Our findings show that being overweight and obese in the pregestational period is associated with adipocyte hypertrophy and specific ATMs populations in VAT.

Matrix Metalloproteinase-3 (MMP-3) Is an Endogenous Activator of the MMP-9 Secreted by Placental Leukocytes: Implication in Human Labor.

BACKGROUND: The activity of matrix degrading enzymes plays a leading role in the rupture of the fetal membranes under normal and pathological human labor, and matrix metalloproteinase-9 (MMP-9) it is considered a biomarker of this event. To gain further insight into local MMP-9 origin and activation, in this study we analyzed the contribution of human placental leukocytes to MMP-9 secretion and explored the local mechanisms of the pro-enzyme activation. METHODS: Placental blood leukocytes were obtained from women at term gestation without labor and maintained in culture up to 72 h. MMP-9 activity in the culture supernatants was determined by zymography and using a specific substrate. The presence of a potential pro-MMP-9 activator in the culture supernatants was monitored using a recombinant biotin-labeled human pro-MMP-9. To characterize the endogenous pro-MMP-9 activator, MMP-1, -3, -7 and -9 were measured by multiplex assay in the supernatants, and an inhibition assay of MMP-9 activation was performed using an anti-human MMP-3 and a specific MMP-3 inhibitor. Finally, production of MMP-9 and MMP-3 in placental leukocytes obtained from term pregnancies with and without labor was assessed by immunofluorescence. RESULTS: Placental leukocytes spontaneously secreted pro-MMP-9 after 24 h of culture, increasing significantly at 48 h (P≤0.05), when the active form of MMP-9 was detected. Culture supernatants activated the recombinant pro-MMP-9 showing that placental leukocytes secrete the activator. A significant increase in MMP-3 secretion by placental leukocytes was observed since 48 h in culture (P≤0.05) and up to 72 h (P≤0.001), when concentration reached its maximum value. Specific activity of MMP-9 decreased significantly (P≤0.005) when an anti-MMP-3 antibody or a specific MMP-3 inhibitor were added to the culture media. Placental leukocytes from term labor produced more MMP-9 and MMP-3 compared to term non-labor cells. CONCLUSIONS: In this work we confirm that placental leukocytes from human term pregnancies are able to secrete large amounts of MMP-9, and that the production of the enzyme it is enhanced by labor. We also demonstrate for the first time that endogenous MMP-3 plays a major role in MMP-9 activation process. These findings support the contribution of placental leukocytes to create the collagenolytic microenvironment that induces the rupture of the fetal membranes during human labor.

[Sperm capacitation with double ejaculate of short interval. A choice in assisted reproduction]. / Capacitación espermática con doble eyaculado de corto intervalo. Una alternativa en reproducción asistida.

In the techniques of assisted reproduction a minimum number of spermatozoa are required, but some patients cannot provide them with one capacitated seminal sample. In this study we investigated if in vitro sperm capacitation of two ejaculates obtained one hour apart from patients with reduced sperm amounts provides an adequate number of spermatozoa. Samples of 75 patients with a low seminal account were processed. They provided one second seminal sample after 60 minutes from the first one. The parameters of the capacitated sample of the first ejaculate (PE) were compared with the parameters of the capacitated sample joining the first and the second ejaculates (PSE). A paired t-test was applied considering a significant value of p < 0.05. Concentration, mobility, total of mobile cells (TCM) and total of mobile cells with normal morphology (ICR) were 31.28 +/-17.65 million/mililiter, 50.45 +/-26.3%, 9.06 +/-7.85 million and 0.64 +/- 0.62 million, respectively in first ejaculate. The respective values in first and second ejaculates were 54.45 +/- 35.06 million/mililiter, 59.39 +/- 25.41%, 17.94 +/- 12.18 million and 1.32 +/- 1.11 million, which represents a significant increase (p<0.05) of 74%, 17%, 98% and 106% respectively. Neither the volume (0.49 +/- 0.037 mililiter versus 0.49 +/- 0.019 mililiter, increase 0%) nor morphology (5.66 +/- 2.76% versus 6.11 +/- 4.88%, increase 7.9%) had a significant change. Sperm capacitation with double ejaculate of short interval must be considered for patients with decreased sperm amounts when deciding the reproductive technique.

Factores que alteran los resultados del análisis computarizado de semen / Computarized semen analysis. Factor affecting results

Objetivo. El presente trabajo tiene como objetivo presentar los datos de la estandarización del analizador automatizado del semen Hamilton-Throrn IVOS 2000. Diseño. Estudio comparativo de las variables de densidad y movilidad espermática usando el método manual (OMS) y el análisis computarizado de semen. Material y método. Posterior a la definición de las condiciones óptimas de lectura, se analizaron de manera cegada 30 muestras seminales que fuero procesadas para análisis de acuerdo con el protocolo de la OMS y por el método automatizado con equipo Hamilton Thorn IVOS 2000. Resultados. La estandarización del método automatizado de semen indica que las lecturas de densidad y movilidad espermática están sujetas a una gran variación que depende de las condiciones en las que se realice el análisis. Las principales fuentes de variación son la frecuencia de adquisición de imágenes y el número de imágenes analizadas. Sin embargo, una vez estandarizadas las condicioens de lectura, el equipo automatizado muestra una buena correlación en la medida de concentración (r= 0.99) y movilidad total (r= 0.98) con el método manual. El coeficiente de variación intraensayo del método automatizado fue de 5.28 por ciento en la lectura de densidad y de 8.65 en la de movilidad total. Conclusiones. El método automatizado de análisis seminal es una herramienta de gran utilidad para un laboratorio de andrología por su reproducibilidad pero es indispensable que cada laboratorio de andrología por su reproducibilidad pero es indispensable que cada laboratorio valide sus mediciones, sobre todo si éstas se utilizan en el área de investigación