Melatonin Action on the Activity of Phagocytes from the Colostrum of Obese Women.

BACKGROUND AND OBJECTIVES: Breastfeeding promotion is an important public health strategy for counter-balancing the negative effects of maternal overweight and obesity. Colostrum contains melatonin, which can attenuate the impacts of excessive maternal weight and boost the infant's immune system. Therefore, the objective of this study was to analyze the effects of melatonin on mononuclear (MN) phagocytes from the colostrum of women with pre-gestational obesity. Materials and Methods: Colostrum samples were collected postpartum from 100 women at a public hospital in São Paulo, Brazil. The donors were divided into two groups: the control group and the high body mass index (BMI) group. Melatonin levels in the colostrum were determined by an ELISA Kit, and the functional activity of MN cells was assessed using the phagocytosis assay by flow cytometry, and reactive oxygen species (ROS), intracellular calcium, and apoptosis were assessed by fluorimetry using a microplate reader. RESULTS: The colostrum of mothers with pre-gestational high BMI exhibited higher melatonin levels (p < 0.05) and lower phagocytosis (p < 0.05) and ROS release (p < 0.05). Superoxide release was similar between the normal and high BMI groups (p > 0.05). Intracellular calcium release and apoptosis were also higher in the high BMI group (p < 0.05). Melatonin levels likely increased the phagocytosis rate and reduced intracellular calcium release and the apoptosis index (p < 0.05). CONCLUSIONS: The results suggest that melatonin is a possible mechanism for maternal-infant protection against obesity and restores the functional activity of colostrum phagocytes in obese mothers.

Effects of Mangaba (Hancornia speciosa) Fruit Extract Adsorbed onto PEG Microspheres in MCF-7 Breast Cancer Cells Co-Cultured with Blood Cells.

Objective: To evaluate the antitumor effects of polyethylene glycol (PEG) microspheres with adsorbed Hancornia speciosa ethanolic extract (HSEE) on blood mononuclear (MN) cells co-cultured with MCF-7 breast cancer cells. Methods: PEG microspheres were adsorbed with HSEE and examined by flow cytometry and fluorescence microscopy. MCF-7 and MN cells obtained from volunteer donors were pre-incubated alone or co-cultured (MN and MCF-7 cells) for 24 h with or without HSEE, PEG microspheres or PEG adsorbed with HSEE (PEG-HSEE). Cell viability, superoxide release and superoxide dismutase were determined. Results: Fluorescence microscopy showed that PEG microspheres were able to absorb HSEE throughout their surface. Irrespective of the treatment, the viability index of MN cells, MCF-7 and their co-culture was not affected. Superoxide release increased in co-cultured cells treated with HSEE, adsorbed or not onto PEG microspheres. In co-cultured cells, SOD levels in culture supernatant increased in the treatment with HSEE, adsorbed onto PEG microspheres or not. Conclusion: HSEE has direct effects on MN cells co-cultured with MCF-7 cells. The results suggest the benefits of Hancornia speciosa fruit consumption by women at risk of breast cancer. In addition, because PEG-HSEE maintained oxidative balance in co-cultured cells, it is a promising alternative for the treatment of tumor cells.

Obesity as an Inflammatory Agent Can Cause Cellular Changes in Human Milk due to the Actions of the Adipokines Leptin and Adiponectin.

Adiponectin and leptin play roles in the hunger response, and they can induce the inflammatory process as the initial mechanism of the innate immune response. It is possible for alterations in the levels of these adipokines to compromise the functional activity of human colostrum phagocytes. Therefore, the objective of this study is to analyze the effects of adiponectin and leptin on colostrum mononuclear (MN) cells. Colostrum was collected from 80 healthy donors, who were divided into two groups: the control group and the high body mass index (BMI) group. MN cells were used to analyze phagocytosis by flow cytometry, and reactive oxygen species (ROS), intracellular calcium, and apoptosis were assessed by fluorimetry using a microplate reader. Adipokines restored the levels of phagocytosis to the high BMI group (p < 0.05), with a mechanism that is action-dependent on the release of ROS and intracellular calcium. However, adiponectin and leptin simultaneously contributed to better microbicidal activity, thus reflecting an increase in the apoptosis level (p < 0.05) in the high BMI group. Probably, the maintenance of the balance between adiponectin and leptin levels enhances the protection and decreases the indices of neonatal infection in the breastfeeding infants of women with high BMI values. Therefore, policies that support pre-gestational weight control should be encouraged.

Cytokine and adipokine are biofactors can act in blood and colostrum of obese mothers.

Breast milk contains bioactive components that contribute to newborn development. However, colostrum may undergo biochemical and immunological changes as a function of maternal overweight and obesity. To investigate this hypothesis, this study determined the levels of hormones and immunological markers in the serum and colostrum of overweight and obese mothers. Colostrum and serum samples were collected from 15 normoweight, 15 overweight, and 15 obese women for determination of leptin, adiponectin, cytokines (TNF-α, IL-6, IL-10), and C-reactive protein (CRP). Obese mothers exhibited higher levels of serum TNF-α, IL-6, and CRP, serum and colostrum leptin and colostrum adiponectin and lower levels of serum adiponectin. Leptin levels in maternal serum and colostrum were positively correlated, as was pre-pregnancy BMI and serum TNF-α, IL-6, CRP, and leptin. Adiponectin levels in colostrum and serum were negatively correlated. The results suggest that obesity changes hormonal and immunological components of maternal serum and colostrum. The modifications can have short-term and long-term effects on newborn development. © 2016 BioFactors, 43(2):243-250, 2017.

Relação do sobrepeso e obesidade materna sobre parâmetros imunológicos, bioquímicos e hormonais do sangue e colostro humano / Influence of overweight and maternal obesity on immunological, biochemical and hormonal parameters of blood and human colostrum

Introdução: A obesidade materna durante a gestação é caracterizada por um estado exagerado de inflamação comparado às gestações mães sem excesso de peso. Existem evidencias de que um delicado balanço de componentes pró e anti-inflamatórios é transferido da mãe para a criança através da amamentação. Não se sabe se a inflamação sistêmica materna que está presente em mulheres obesas influencia no perfil de citocinas e hormônios do sangue e colostro. Objetivos: Avaliar os perfis de citocinas TH1, TH2, TH3, TH17 no sangue e colostro de mães com excesso massa corporal, e verificar os parâmetros bioquímicos, imunológicos e hormonais do sangue e colostro humano. Método: Foram avaliadas 68 mães, assim distribuídas: eutróficas (n=25), sobrepeso (n=24) e obesidade (n=19). As concentrações de glicose, triglicérides e colesterol total foram mensuradas por método colorimétrico enzimático, e de proteínas totais por método colorimétrico. Teor de gordura e calorias do colostro foram obtidos por método do Crematócrito. Os hormônios leptina e adiponectina foram quantificados por método imunoenzimático de ELISA. Imunoglobulinas, proteínas do complemento (C3 e C4) e PCR por turbidimetria. As concentrações de citocinas foram avaliadas por Citometria de fluxo. Resultados: Observou-se maiores níveis de glicose, teor de gordura, calorias e IgA no colostro de mães obesas.

Introduction: Maternal obesity during pregnancy is characterized by an exaggerated state of inflammation compared to mothers without excess weight. There is evidence that a delicate balance of proinflammatory and anti-inflammatory compounds is transferred from mother to infant via breastfeeding. It is unknown whether the maternal systemic inflammation that is present in obese women influences the cytokine levels of colostrum. Objective: To evaluate the cytokine profile Th1, TH2, TH3, TH17 of blood and colostrum of overweight and obese mothers, and verify biochemical, hormonal and immunological parameters of human blood and colostrum. Methods: Were evaluated 68 mothers that were divided according to their BMI status: lean (n=25), overweight (n=24) and obese (n=19). The concentrations of glucose, triglycerides and total cholesterol were measured by colorimetric enzymatic method, and total proteins by colorimetric method. Fat content and calories of colostrum were obtained by Creamatocrit method. The hormones leptin and adiponectin were measured by ELISA technique. Immunoglobulins and complement protein (C3 and C4) and PCR by Turbidimetric method. Concentrations of cytokines were assessed by flow cytometry. Results: We observed higher glucose levels, fat content, calories and IgA in colostrum of obese mothers. Serum concentrations of triglycerides, cholesterol, IgA, CRP, C3 and C4 complement proteins were significantly higher in obese mothers.

Changes in the biochemical and immunological components of serum and colostrum of overweight and obese mothers.

BACKGROUND: Obesity in pregnancy is associated with systemic inflammation, immunological changes and adverse maternal-fetal outcomes. Information on the association between maternal obesity and breast milk composition is scarce. This study describes changes and relationships between biochemical and immunological parameters of colostrum and serum of overweight and obese women. METHODS: Colostrum and blood samples were collected from 25 normal weight, 24 overweight and 19 obese women for determination of glucose, total protein, triglycerides, cholesterol, immunoglobulins, complement proteins (C3 and C4), fat and calorie content and C-reactive protein (CRP). RESULTS: Glucose was higher in colostrum of obese women (p = .002). In normal weight and obese women, total protein content was higher in colostrum than in serum (p = .001). Serum triglycerides (p = .008) and cholesterol (p = .010) concentrations were significantly higher in overweight and obese women than in their normal weight counterparts, but in colostrum their concentrations were similar across the three groups. Secretory IgA (sIgA) in colostrum and IgA in serum concentrations were significantly higher (p = .001) in overweight and obese mothers, whereas IgG and IgM concentrations did not vary among the groups (p = .825). Serum C3 (p = .001) and C4 (p = .040) concentrations were higher in obese women. No differences in colostrum complement proteins were detected among the groups. Calorie content (p = .003) and fat (p = .005) concentrations in colostrum and serum CRP (p = .002) were higher in obese women. CONCLUSIONS: The results corroborate the hypothesis that colostrum of overweight and obese women undergoes biochemical and immunological changes that affect its composition, namely increasing glucose concentrations, calorie content, fat and sIgA concentrations.

Human colostral phagocytes eliminate enterotoxigenic Escherichia coli opsonized by colostrum supernatant.

BACKGROUND: Several elements in colostrum and human milk, including antibodies and nonspecific factors with bactericidal and antiviral activity, may play an important anti-infectious and protective role. In developing countries, enterotoxigenic Escherichia coli (ETEC) is the main etiological agent of diarrhea in low-socioeconomic level children. In the present work, we studied the functional activity of mononuclear (MN) and polymorphonuclear (PMN) phagocytes of human colostrum against ETEC, as well as the interactions between these cells and colostral or serum opsonins. METHODS: Colostrum samples were collected from 33 clinically healthy women between 48 and 72 hours postpartum. We verified superoxide release in colostral MN and PMN using cytochrome C reduction methods, phagocytosis, and bactericidal activity using acridine orange methods and superoxide dismutase (SOD) in the colostrum supernatants. RESULTS: Colostral MN and PMN phagocytes exposed to ETEC opsonized with colostrum supernatants caused a significant increase (p<0.05) in superoxide release. Phagocytosis by colostral PMN cells increased significantly (p<0.5) when the phagocytes were incubated with both sources of opsonins (sera and colostrum). Increases in superoxide release in the presence of opsonized bacteria triggered the bactericidal activity of the phagocytes. Phagocyte treatment with SOD decreased their ability to eliminate ETEC. Colostrum supernatant had higher SOD concentrations (p<0.05) compared with normal human sera. CONCLUSIONS: These results suggest that the ability of phagocytes to eliminate ETEC depends on the activation of cellular oxidative metabolism; moreover, activation of colostral phagocytes is likely an additional breast-feeding protection mechanism against intestinal infections in infants.