Sandra A. Reza-López

High folate gestational and post-weaning diets alter hypothalamic feeding pathways by DNA methylation in Wistar rat offspring.

Excess vitamins, especially folate, are consumed during pregnancy but later-life effects on the offspring are unknown. High multivitamin (10-fold AIN-93G, HV) gestational diets increase characteristics of metabolic syndrome in Wistar rat offspring. We hypothesized that folate, the vitamin active in DNA methylation, accounts for these effects through epigenetic modification of food intake regulatory genes. Male offspring of dams fed 10-fold folate (HFol) diet during pregnancy and weaned to recommended vitamin (RV) or HFol diets were compared with those born to RV dams and weaned to RV diet for 29 weeks. Food intake and body weight were highest in offspring of HFol dams fed the RV diet. In contrast, the HFol pup diet in offspring of HFol dams reduced food intake (7%, p = 0.02), body weight (9%, p = 0.03) and glucose response to a glucose load (21%, p = 0.02), and improved glucose response to an insulin load (20%, p = 0.009). HFol alone in either gestational or pup diet modified gene expression of feeding-related neuropeptides. Hypomethylation of the pro-opiomelanocortin (POMC) promoter occurred with the HFol pup diet. POMC-specific methylation was positively associated with glucose response to a glucose load (r = 0.7, p = 0.03). In conclusion, the obesogenic phenotype of offspring from dams fed the HFol gestational diet can be corrected by feeding them a HFol diet. Our work is novel in showing post-weaning epigenetic plasticity of the hypothalamus and that in utero programming by vitamin gestational diets can be modified by vitamin content of the pup diet.

High multivitamin intake by Wistar rats during pregnancy results in increased food intake and components of the metabolic syndrome in male offspring

The effect of high multivitamin intake during pregnancy on the metabolic phenotype of rat offspring was investigated. Pregnant Wistar rats (n=10 per group) were fed the AIN-93G diet with the recommended vitamin (RV) content or a 10-fold increase [high vitamin (HV) content]. In experiment 1, male and female offspring were followed for 12 wk after weaning; in experiment 2, only males were followed for 28 wk. Body weight (BW) was measured weekly. Every 4 wk, after an overnight fast, food intake over 1 h was measured 30 min after a gavage of glucose or water. An oral glucose tolerance test was performed every 3-5 wk. Postweaning fasting glucose, insulin, ghrelin, glucagon-like peptide-1, and systolic blood pressure were measured. No difference in BW at birth or litter size was observed. Food intake was greater in males born to HV dams (P<0.05), and at 28 wk after weaning, BW was 8% higher (P<0.05) and fat pad mass was 27% higher (P<0.05). Food intake reduction after the glucose preload was nearly twofold less in males born to HV dams at 12 wk after weaning (P<0.05). Fasting glucose, insulin, and ghrelin were 11%, 62%, and 41% higher in males from HV dams at 14 wk after weaning (P<0.05). Blood glucose response was 46% higher at 23 wk after weaning (P<0.01), and systolic blood pressure was 16% higher at 28 wk after weaning (P<0.05). In conclusion, high multivitamin intake during pregnancy programmed the male offspring for the development of the components of metabolic syndrome in adulthood, possibly by its effects on central mechanisms of food intake control.

Flaxseed combined with low-dose estrogen therapy preserves bone tissue in ovariectomized rats.

Objective: Flaxseed is rich in lignans and &agr;-linolenic acid, compounds that may promote healthy skeletons. Many postmenopausal women consume complementary health products such as flaxseed or its components in addition to pharmacological agents such as low-dose estrogen therapy for additional support for menopausal symptoms and related conditions. However, their combined effect on bone health is unknown. The aim of this study was to determine the effects of 10% dietary flaxseed, low-dose estrogen therapy, or their combination on bone mineral density, biomechanical strength, and skeletal fatty acid composition in an ovariectomized rat model of postmenopausal osteoporosis. Methods: Ovariectomized rats received (1) basal diet (negative control), (2) 10% flaxseed, (3) low-dose estrogen implant (13 &mgr;g, 90 day release), or (4) flaxseed + low-dose estrogen implant for 12 weeks. A sham-operated group was included as a positive control. Bone mineral density, biomechanical strength, and fatty acid composition were measured at multiple skeletal sites. Results: Flaxseed + low-dose estrogen therapy resulted in the highest bone mineral density and peak load at the lumbar vertebrae, with no effect on bone mineral density or strength in the tibia and femur. Flaxseed and flaxseed + low-dose estrogen therapy resulted in significantly higher relative levels of &agr;-linolenic acid and eicosapentaenoic acid and lower levels of linoleic acid, arachidonic acid, and n-6/n-3 ratio in the lumbar vertebrae and tibia compared with all other groups. Conclusion: Flaxseed + low-dose estrogen therapy provides the greatest protection against ovariectomy-induced bone loss at the lumbar vertebrae. Moreover, this study is the first to demonstrate that flaxseed, rich in &agr;-linolenic acid, alters fatty acid composition in the ovariectomized rat skeleton.

Seizure resistance in fat-1 transgenic mice endogenously synthesizing high levels of omega-3 polyunsaturated fatty acids

n‐3 polyunsaturated fatty acids (PUFA), derived from marine oils, have been shown to protect against various neurological diseases. However, very little is known about their potential anticonvulsant properties. The objective of the present study was to determine whether enrichment of brain lipids with n‐3 PUFA inhibits seizures induced by pentylenetetrazol. We demonstrate that increased brain levels of n‐3 PUFA in transgenic fat‐1 male mice, which are capable of de novo synthesis of n‐3 PUFA from n‐6 PUFA, increases latency to seizure onset by 45%, relative to wildtype controls (p = 0.08). Compared with wildtype littermates, transgenic fat‐1 mice have significantly (p < 0.05) higher levels of docosahexaenoic acid and total n‐3 PUFA in brain total lipid extracts and phospholipids. Levels of brain docosahexaenoic acid were positively correlated to seizure latency (p < 0.05). These findings demonstrate that n‐3 PUFA have anticonvulsant properties and suggest the possibility of a novel, non‐drug dietary approach for the treatment of epilepsy.

Effects of exposure to pesticides during pregnancy on placental maturity and weight of newborns: A cross-sectional pilot study in women from the Chihuahua State, Mexico

It is known that pesticides cross the placental barrier and can cause alterations in the development of placental structures resulting in adverse effects in reproduction. The objectives of this study were to investigate the effects of pesticide exposure during pregnancy on placental maturity and to evaluate the relationship between placental maturity, gestational age and birth weight. We collected the placentas from singleton pregnancies from women exposed (n = 9) and non-exposed (n = 45 full-term and n = 31 preterm) to pesticides as evaluated geographically, by questionnaire and by acetylcholinesterase levels. Placental morphometry from the central and peripheral regions was examined by microscopy and staining with hematoxylin and eosin. The placental maturity index (PMI) was estimated by dividing the number of epithelial plates in terminal villi to their thickness in 1 mm2 of the placental parenchyma. Gestational age, birth weight and the following characteristics of the mother were also recorded: pre-pregnancy body mass index, weight gain during pregnancy and hemoglobin concentrations. Birth weight and the gestational age were correlated with PMI (r = .54 and r = .44, respectively; p < .01). Pesticide exposure was associated with a higher PMI (beta = 7.38, p = .01) after adjusting by variables related to placental maturity. In conclusion, the results suggest a relationship between prenatal exposure to pesticides and placental maturity and may potentially affect the nutrient transport from the mother to the fetus.

High vitamin intake by Wistar rats during pregnancy alters tissue fatty acid concentration in the offspring fed an obesogenic diet

Diet during pregnancy affects the long-term health of the offspring. Vitamins are known to modulate lipid metabolism, which may be reflected in tissue fatty acid (FA) concentrations. The objective of this study was to investigate the effect of high vitamin intake during pregnancy on tissue FA concentration of the offspring. Wistar rats were fed an AIN-93G diet with either the recommended vitamin or 10-fold higher amounts (HV) during pregnancy. Afterward, offspring were weaned onto an obesogenic diet. Liver, quadriceps, adipose, and brain were collected over 48 weeks. Fatty acid concentration of tissue total lipids was analyzed by gas chromatography. At birth, the liver from HV offspring was higher in monounsaturated, stearic, and arachidonic acids. At weaning, the liver from HV offspring was higher in stearic and oleic acids; and in adipose tissue, n-6 and n-3 FAs were lower only in the male HV offspring (P < .05). At 12 weeks, HV offspring had higher concentrations of total fat, saturates, monounsaturates, and n-6 FA in muscle (P < .05), but not in other tissues. At 48 weeks, gestational diet did not affect tissue total lipid FA concentrations; but differences remained in specific tissue phospholipids species. Liver phospholipids from HV offspring were lower in monounsaturates and n-6 FA. Brain phosphatidylethanolamine was higher in oleic, n-6 FA, and docosahexaenoic acid in the HV offspring. Phosphatidylinositol was lower in saturates, monounsaturates, arachidonic, and docosahexaenoic acids only in HV female offspring. These observations demonstrate that high vitamin intake during pregnancy has short- and long-term effects on tissue FA concentration in the offspring.

High Folic Acid Intake during Pregnancy Lowers Body Weight and Reduces Femoral Area and Strength in Female Rat Offspring

Rats fed gestational diets high in multivitamin or folate produce offspring of altered phenotypes. We hypothesized that female rat offspring born to dams fed a gestational diet high in folic acid (HFol) have compromised bone health and that feeding the offspring the same HFol diet attenuates these effects. Pregnant rats were fed diets with either recommended folic acid (RFol) or 10-fold higher folic acid (HFol) amounts. Female offspring were weaned to either the RFol or HFol diet for 17 weeks. HFol maternal diet resulted in lower offspring body weights (6%, P = 0.03) and, after adjusting for body weight and femoral length, smaller femoral area (2%, P = 0.03), compared to control diet. After adjustments, HFol pup diet resulted in lower mineral content (7%, P = 0.01) and density (4%, P = 0.002) of lumbar vertebra 4 without differences in strength. An interaction between folate content of the dam and pup diets revealed that a mismatch resulted in lower femoral peak load strength (P = 0.01) and stiffness (P = 0.002). However, the match in folate content failed to prevent lower weight gain. In conclusion, HFol diets fed to rat dams and their offspring affect area and strength of femurs and mineral quantity but not strength of lumbar vertebrae in the offspring.

Increasing vitamin A in post-weaning diets reduces food intake and body weight and modifies gene expression in brains of male rats born to dams fed a high multivitamin diet.

High multivitamin gestational diets (HV, 10-fold AIN-93G levels) increase body weight (BW) and food intake (FI) in rat offspring weaned to a recommended multivitamin (RV), but not to a HV diet. We hypothesized that high vitamin A (HA) alone, similar to HV, in post-weaning diets would prevent these effects of the HV maternal diet consistent with gene expression in FI and reward pathways. Male offspring from dams fed HV diets were weaned to a high vitamin A (HA, 10-fold AIN-93G levels), HV or RV diet for 29 weeks. BW, FI, expression of genes involved in regulation of FI and reward and global and gene-specific DNA methylation of pro-opiomelanocortin (POMC) in the hypothalamus were measured. Both HV and HA diets slowed post-weaning weight gain and modified gene expression in offspring compared to offspring fed an RV post-weaning diet. Hypothalamic POMC expression in HA offspring was not different from either HV or RV, and dopamine receptor 1 was 30% (P<.05) higher in HA vs. HV, but not different from RV group. Hippocampal expression of serotonin receptor 1A (40%, P<.01), dopamine receptor 2 (40%, P<.05) and dopamine receptor 5 (70%, P<.0001) was greater in HA vs. RV fed pups and is 40% (P<.01), 50% (P<.05) and 40% (P<.0001) in HA vs. HV pups, respectively. POMC DNA methylation was lower in HA vs. RV offspring (P<.05). We conclude that high vitamin A in post-weaning diets reduces post-weaning weight gain and FI and modifies gene expression in FI and reward pathways.

Flaxseed does not antagonize the effect of ultra-low-dose estrogen therapy on bone mineral density and biomechanical bone strength in ovariectomized rats.

A previous study showed that flaxseed (FS) combined with low-dose (LD) estrogen therapy, resembling LD transdermal estrogen therapy in postmenopaual women, inhibited loss of bone mineral density (BMD), bone mineral content (BMC), and strength in lumbar vertebrae in ovariectomized rats. Whether FS combined with an even lower dose of estrogen is effective at preserving bone or whether FS interferes with the effect of this lower dose of estrogen is unknown. Thus, this study determined whether an ultra-low-dose (ULD) estrogen therapy, half the dose previously studied, in combination with FS preserved bone mass and strength in the lumbar vertebrae in ovariectomized rats. Rats were treated for 12 wk with (1) basal diet (BD) (ovariectomized control), (2) BD + ULD estrogen implant, or (3) BD containing 10% FS + ULD estrogen implant. A sham-operated control group was fed BD. Unlike ULD, FS + ULD attenuated loss of BMD and strength at the lumbar vertebrae and BMD in femurs and tibias. FS + ULD resulted in higher percentages of n-3 fatty acids including alpha-linolenic acid and eicosapentaenoic acid and lower percentages of n-6 fatty acids including linoleic acid compared to all other groups. Differences in fatty acid composition at the lumbar vertebrae and tibia were significantly related to BMD, BMC, and strength. No treatment-induced effects on uterus weight were observed, but histological analyses are needed to confirm safety. In conclusion, FS did not antagonize the activity of ULD, and their combination attenuated the loss of BMD and strength at the lumbar vertebrae, which was associated with differences in bone fatty acid composition.

Body Composition of Women with Newborns Who Are Small for Gestational Age

Background: The relationship between maternal and placental hemodynamic disorders and fetal growth is well known, but few studies have evaluated a link between maternal extracellular water (EW) and newborn birth weight. Objective: To identify the characteristics of body composition (BC) of women with small for gestational age (SGA) newborns, and to determine the relationship between maternal EW and birth weight of the baby. Methods: We studied maternal BC using multifrequency bioelectric impedance in the second and third trimesters of pregnancy. Newborns with weight below the 10th percentile were classified as SGA; those with weights between the 10th and 90th percentiles as appropriate for gestational age (AGA), and large for gestational age (LGA) were those with weights above the 90th percentile. Results: We studied 460 women and their BC varied depending on whether their newborns were SGA, AGA or LGA. EW was lower in the mothers of SGA (11 ± 2 l) compared to AGA (12 ± 3 l) newborns (p < 0.01). We identified a significant relationship (p < 0.01) between maternal EW in the second trimester and the weight of the newborn, β = 43 (95% CI 27–58). Conclusion: BC of women whose newborns are SGA differs significantly from that of women whose newborns are AGA, a result which suggests that the mothers of SGA infants may have a disordered hemodynamic state during the second trimester of pregnancy.