Pup Weight Research Articles

Hexamoll® DINCH: Lack of in vivo evidence for obesogenic properties

Hexamoll® DINCH is an important alternative to phthalate plasticizers. Although regulatory reviews have not identified any potential hazards even in sensitive populations, an in vitro study by Campioli et al. (2015) suggested Hexamoll® DINCH might alter fat storage in adipocytes resulting in obesity. To evaluate this hypothesis, data from studies with Hexamoll® DINCH were reviewed for evidence of deposition in fat, changes in body weight, or changes in serum chemistry reflecting altered metabolic status. Body weights of F1 and F2 pups in a two-generation study did not differ from controls even at 1000 mg Hexamoll® DINCH/kg body weight. Mean relative liver weights from the 1000 and 300 mg/kg bw groups were increased, but without histopathologic changes. Triglyceride and cholesterol levels in serum were not affected. In addition, subchronic and chronic studies in rats did not give evidence of an obesogenic effect. Radioactivity from 20 or 1000 mg/kg bw 14C-labelled Hexamoll® DINCH dosed orally remained 2–3 times longer in adipose tissue than in well-perfused tissues; however, levels were 20–500% below other tissues at 1 and 8 h post dosing. Radioactivity concentrations in organs and tissues excluding the GI tract declined rapidly and continuously, and decreased in parallel to the concentration in plasma during the following 20 h. Both, initial and terminal half-lives of radioactivity concentration do not indicate a potential for accumulation. Furthermore, a metabolomic comparison of Hexamoll® DINCH with DEHP and other phthalates shows complete separation of the metabolomic profile of these two chemical classes, meaning that their effects on the body and the body’s reaction to the substance are different. Hence, comprehensive in vivo data do not show any evidence of Hexamoll® DINCH altering fat metabolism or having obesogenic properties.

Administration of Lactobacillus plantarum Lp62 to dam rats at the end of delivery and during lactation affects TGF-β1 level and nutritional milk composition, and body weight of pups.

Lactobacillus plantarum Lp62 is a lactic acid bacteria strain that has been isolated from cocoa beans and exhibited probiotic potential. The influence of oral administration of L. plantarum Lp62 on the growth of rat's pups; on yield, cytokines and milk composition was studied. Lactobacillus plantarum Lp62 is a lactic acid bacteria strain that has been isolated from cocoa beans. It was administered daily by gavage to Wistar rats (n = 8), from the 7th day before delivery and for 20 days during lactation, in a concentration of 1.44 × 109 CFU/rat. The dam and pups were weighed and milk was collected at 12th and 19th day for determination of protein, triglycerides, cholesterol and lactose by colorimetric assays. TGF-β1 milk levels were analyzed by ELISA. The mammary glands of rats were removed for histological analysis. To detect statistical differences between the groups, tests of mean differences at a significance level of 5% was performed. Supplementation with L. plantarum L62 resulted in significant higher weight of pups (p < 0.05), with similar weight on dams (p > 0.05). The milk yield was not altered by L. plantarum treatment, but the levels of protein, triglycerides and cholesterol were increased (p < 0.05), with no difference in lactose concentration (p > 0.05). Levels of TGF-β1 were higher in the milk of L. plantarum treatment (p < 0.05). The treatment of dams at the end of pregnancy and lactation with L. plantarum Lp62 increased nutritional content of milk, probably contributing to the higher weight of the pups. The higher levels of TGF-β1 in the milk, could promote immune benefits to the pups. Further studies in this field are needed to prove the potential use of L. plantarum Lp62 as a probiotic.

The Pyramid Counteracts Chronic Prenatal Restraint-Stress Effects on the Milestones, Anthropometry, and Body, Brain and Adrenal Gland Weights of Pups in Rats

Prenatal exposure to chronic stress during critical periods of foetal development produces depression, attention and learning deficits, hormonal imbalances and affects the brain. The effect of prenatal restraint-stress on the postnatal developmental milestones, anthropometric measurements, and the body, brain and adrenal gland weights of the pups were examined and compared with the un-restrained control and the restrained group under the pyramid at postnatal day 10 and 21. Pregnant rats were restrained (9h/day) from gestation day 7 until parturition. Results showed significant delay in the milestones by one day in the restraint control (RC) compared to the unrestrained normal control (NC), while pups of the restrained pyramid (RP) group did not show the delay. Significant decreases in the anthropometric measurements, body and brain weights in RC group were observed at both postnatal days, while the RP group results matched with the NC group. Significant increase in the adrenal weights was found in the RC group compared to NC group and not the RP group. Results suggest prenatal restraint-stress definitely hampers the developmental milestones, anthropometric measurements, and body and brain weights of the young offspring. Results suggest, pyramid environment counteracts and protects the deleterious effects of chronic prenatal stress. Keywords: Anthropometry, Milestones, Prenatal stress, Pyramid

Compromised JMJD6 Histone Demethylase Activity Affects VHL Gene Repression in Preeclampsia.

The von Hippel Lindau (VHL) protein is a key executor of the cellular hypoxic response that is compromised in preeclampsia, a serious disorder complicating 5% to 7% of pregnancies. To date, the mechanisms controlling VHL gene expression in the human placenta remain elusive. We examined VHL epigenetic regulation in normal pregnancy and in preeclampsia, a pathology characterized by placental hypoxia. Placentae were obtained from early-onset preeclampsia (n = 56; <34 weeks of gestation) and late-onset preeclampsia (n = 19; ≥34 weeks of gestation). Placentae from healthy normotensive age-matched preterm control (n = 43) and term control (n = 23) pregnancies were included as controls. We measured the activity of Jumonji domain containing protein 6 (JMJD6), a ferrous iron (Fe2+)- and oxygen-dependent histone demethylase, and examined its function in the epigenetic control of VHL. JMJD6 regulates VHL gene expression in the human placenta. VHL downregulation in preeclampsia is dependent on decreased JMJD6 demethylase activity due to hypoxia and reduced Fe2+ bioavailability. Chromatin immunoprecipitation assays revealed decreased association of JMJD6 and its histone targets with the VHL promoter. Findings in preeclampsia were corroborated in a murine model of pharmacological hypoxia using FG-4592. Placentae from FG-4592-treated mice exhibited reduced VHL levels, accompanied by placental morphological alterations and reduced pup weights. Notably, Fe2+ supplementation rescued JMJD6 histone demethylase activity in histone from E-PE and FG-4592-treated mice. Our study uncovers epigenetic regulation of VHL and its functional consequences for altered oxygen and iron homeostasis in preeclampsia.

A new model for fetal programming: maternal Ramadan-type fasting programs nephrogenesis.

The effect of maternal Ramadan-type fasting (RTF) on the outcome of pregnancy, kidney development and nephron number in male rat offspring was investigated in current study. Pregnant rats were given food and water ad libitum during pregnancy (control) or restricted for 16 h per day (RTF). Kidney structure was examined during fetal life, at birth, and in early and late adulthood. Maternal body weight, food intake, relative food intake and plasma glucose levels were significantly lower (P<0.001) in the RTF group. Litter and pup weights also were significantly lower (P<0.05) in the RTF group at birth, with no difference in the litter size. The RTF group had a longer gestation, delayed nephrogenesis with less well-differentiated glomeruli, more connective tissue, fewer medullary rays, an increase in the nephrogenic zone/cortical zone ratio, and significant increase (P<0.001) in kidney apoptosis at birth. On the other hand, maternal fasting reduced nephron number (by ~31%) with unchanged kidney and total glomerular volumes. Mean glomerular volume was significantly higher in RTF offspring. Assessment of renal structure revealed mild glomerulosclerosis with enlarged lobulated glomeruli in the renal cortex and high interstitial fibrosis in the medulla of RTF kidneys. Taken together, gestational fasting delays nephrogenesis and reduces nephron number in the kidneys of the offspring, that could be partially owing to increased apoptosis.

The Potential Therapeutic Role of Peroxisome ProliferatorActivated Receptors Agonist in Preeclamptic Pregnant Rats.

To investigate the role of peroxisome proliferator-activated receptor (PPAR)-&gamma; agonist on renal functions and blood levels of vasoregulatory peptides by administering rosiglitazone-a PPAR-&gamma; agonist- to nitric oxide (NO) synthase lacking pregnant rats as a creature model of preeclampsia. Experimental study. Medical College at King Khaled University Hospital, Riyadh, Saudi Arabia, from July 2016 to May 2017. One hundred female rats were divided into four equal groups; virgin non-pregnant, pregnant salinereceived, pregnant treated with NG-nitro-L-arginine-methyl-ester (L-NAME) to produce preeclampsia and the last group received L-NAME+rosiglitazone. Some renal function tests and blood level of vasoregulatory factors were measured on day 13 and day 20 of pregnancy. Pup weight was also measured. L-NAME treated rats exhibited all classic hallmarks of preeclampsia. Rosiglitazone treatment from days 14 to 20 ameliorated hypertension, improved renal function, decreased endothelin-1 (ET-1), angiotensin II (Ag II) and interleukin (IL-6), while it increases NO level and pup weight. Administration of rosiglitazone can improve some of the pathophysiological trademarks related with preeclampsia and may prove useful for the prevention of the development and progression of this condition. However, human trials are necessary to prove this hypothesis.

Identification of early indicators of altered metabolism in normal development using a rodent model system.

ABSTRACTAlthough the existence of a close relationship between the early maternal developmental environment, fetal size at birth and the risk of developing disease in adulthood has been suggested, most studies, however, employed experimentally induced intrauterine growth restriction as a model to link this with later adult disease. Because embryonic size variation also occurs under normal growth and differentiation, elucidating the molecular mechanisms underlying these changes and their relevance to later adult disease risk becomes important. The birth weight of rat pups vary according to the uterine horn positions. Using birth weight as a marker, we compared two groups of rat pups – lower birth weight (LBW, 5th to 25th percentile) and average birth weight (ABW, 50th to 75th percentile) – using morphological, biochemical and molecular biology, and genetic techniques. Our results show that insulin metabolism, Pi3k/Akt and Pparγ signaling and the genes regulating growth and metabolism are significantly different in these groups. Methylation at the promoter of the InsII (Ins2) gene and DNA methyltransferase 1 in LBW pups are both increased. Additionally, the Dnmt1 repressor complex, which includes Hdac1, Rb (Rb1) and E2f1, was also upregulated in LBW pups. We conclude that the Dnmt1 repressor complex, which regulates the restriction point of the cell cycle, retards the rate at which cells traverse the G1 or G0 phase of the cell cycle in LBW pups, thereby slowing down growth. This regulatory mechanism mediated by Dnmt1 might contribute to the production of small-size pups and altered physiology and pathology in adult life.

Effect of Continuous Trio Breeding Compared with Continuous Pair Breeding in ‘Shoebox’ Caging on Measures of Reproductive Performance in Estrogen Receptor Knockout Mice

Some performance standards for continuous trio breeding in 'shoebox' cages for inbred stocks and outbred strains ofmice challenge the minimum floor space recommendations in the 8th edition of the Guide for the Care and Use of LaboratoryAnimals. In our study, we evaluated whether continuous trio breeding could be successfully applied to a breeding colonyof genetically engineered mice housed in shoebox cages with a floor area of 67.6 in2. Mice heterozygous for genetically engineered mutations to estrogen receptors and their wildtype counterparts were continuously bred as trios or pairs. Confounding environmental factors were controlled through standardized husbandry practices and husbandry, and all mice were bred simultaneously to control for temporal factors. Several measures of reproductive performance-including number of littersper female, production index, interlitter interval, litter size at birth, litter size at weaning, weaning rate, and body weight of pups at weaning- were evaluated over approximately 6 mo. Regardless of genotype, interlitter interval, litter size at birth, and litter size at weaning were significantly lower for trio-bred mice than for pair-bred mice. In addition, significant interactions emerged between genotype and breeding strategy for these reproductive measures. Furthermore, significant differencesbetween genotypes occurred for interlitter interval and weaning rate, regardless of breeding strategy. Underlying mechanismsto account for effects of genotype on interlitter interval and the interaction of genotype with breeding strategy were unclearbut may reflect effects of overcrowding and reproductive suppression.

Effect of Folic Acid Supplementation and/or its Absence on Body Weight of Wistar Rats in Pregnancy and in their Offspring

Introduction: Foetal birth weight is the best predictor of pregnancy outcome as the growth restriction has been associated with high morbidity and mortality. The need of Folic Acid (FA) supplementation is essential during the period of pregnancy to prevent untoward effects on pregnancy related complication in mother and to prevent birth defects in foetus. Aim: To compare the effects of FA supplementation and FA absent diets during the pregnancy on maternal and their offspring body weight. Materials and Methods: This was a prospective study comprising of 18 Wistar albino rats procured from the Animal house, Manipal Academy of Higher Education, Karnataka, India, after the institutional ethical committee clearance. Rats were randomly divided into three groups and given different diets: Control group with normal diet, group with FA supplementation diet, and group with FA absent diet (with added succinyl sulfathiazole) for five weeks. Later the female adult Wistar rats could mate with male rats of their own strain and allowed the female to complete their pregnancy. After the delivery, delivered pups were counted for each rat and gross malformation in each pup were noted. Immediately after the delivery, all the pups born to each rat were weighed individually in digital weighing machine. Results: Among the pregnant dams the body weight gain was observed more in control group with an average weight of 40.56 gm, followed by diet with absent FA (32 gm) and least in rats fed with FA supplementation diets (31 gm). Average number of pups born to pregnant dams fed with FA supplementation, FA absent diet, and in normal diet was found to be 8, 8, and 9 respectively. Regarding the birth weight of pups born to pregnant dams fed with FA supplementation the average weight was found to be 6.01-5.09 gm, with normal diet it was 5.51-5.08 gm, and least weight was observed in pups born with FA absent diet (4.74-3.31 gm). Conclusion: The present study concluded that the FA status in pregnancy is inversely proportional to the prenatal death in pups but in offspring’s born to mothers fed with FA supplementation, average body weight was more than those born to mothers fed with FA absent diet and with control groups. However, this needs further confirmation as the tests were done on limited samples.

Hydrogen Sulfide Ameliorates Developmental Impairments of Rat Offspring with Prenatal Hyperhomocysteinemia.

Maternal high levels of the redox active amino acid homocysteine—called hyperhomocysteinemia (hHCY)—can affect the health state of the progeny. The effects of hydrogen sulfide (H2S) treatment on rats with maternal hHCY remain unknown. In the present study, we characterized the physical development, reflex ontogeny, locomotion and exploratory activity, muscle strength, motor coordination, and brain redox state of pups with maternal hHCY and tested potential beneficial action of the H2S donor—sodium hydrosulfide (NaHS)—on these parameters. Our results indicate a significant decrease in litter size and body weight of pups from dams fed with methionine-rich diet. In hHCY pups, a delay in the formation of sensory-motor reflexes was observed. Locomotor activity tested in the open field by head rearings, crossed squares, and rearings of hHCY pups at all studied ages (P8, P16, and P26) was diminished. Exploratory activity was decreased, and emotionality was higher in rats with hHCY. Prenatal hHCY resulted in reduced muscle strength and motor coordination assessed by the paw grip endurance test and rotarod test. Remarkably, administration of NaHS to pregnant rats with hHCY prevented the observed deleterious effects of high homocysteine on fetus development. In rats with prenatal hHCY, the endogenous generation of H2S brain tissues was lower compared to control and NaHS administration restored the H2S level to control values. Moreover, using redox signaling assays, we found an increased level of malondialdehyde (MDA), the end product of lipid peroxidation, and decreased activity of antioxidant enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GPx) in the brain tissues of rats of the hHCY group. Notably, NaHS treatment restored the level of MDA and the activity of SOD and GPx. Our data suggest that H2S has neuroprotective/antioxidant effects against homocysteine-induced neurotoxicity providing a potential strategy for the prevention of developmental impairments in newborns.

270: Commensal microbes confer protection against Zika virus infection in a murine gnotobiotic model of congenital Zika syndrome

Recent findings reveal that commensal microbiota may play a role in protection against viral infections. For example, bacteria isolated from the human stomach confer resistance against rotavirus infection in mice. Vertical transmission of ZIKV has been shown cause IUGR in mice. Here we sought to determine the contribution of commensal bacteria in protecting against ZIKV infection, vertical transmission, and IUGR. Current pandemic ZIKV was isolated from a traveler returning from Central America and propagated in Vero cells (ZIKV HN16). Pregnant germ-free (GF) and conventional (CONV) Swiss-Webster mice were intravenously inoculated with 104 PFU of second passage ZIKV (n=11) or a mock (n=10) injection on embryonic day 6 (e6), then daily until day e9 (Figure 1). Mice (n=2) were given anti-IFNI1 ab prior to and after infection as positive infection controls. On e18, Cesareans were performed and mice assessed for intrauterine reabsorption and demise as well as embryonic pup weight. Tissues were assessed for ZIKV infection by TaqMan viral qPCR assay. Inoculated mice were asymptomatic based on daily assessment of mobility and paralysis. Overall, GF-ZIKV pups had a greater rate of in utero demise with ZIKV infection (p< 0.01), while there was no difference for conventional controls (Fig 2A). In GF-ZIKV dams, ZIKV was detected in 3 of 4 spleens assessed, but only detected in 1 of 4 spleens from GF-ZIKV mice, and with 100-fold less viral load (B). The uterus was positive for ZIKV in 75% (3/4) GF-ZIKV dams in comparison to 50% (2/4) CONV-ZIKV dams, also with 100-fold less viral load (B). Of the mice positive for ZIKV in the uterus, 3 of 4 had associated in utero demise, while all pregnancies with undetectable levels of ZIKV in the uterus had no incidences of in utero demise (Fig A/B?). Pups of Zika infected conventional mothers showed no difference in fetal weight compared to mock, while germ free mothers were 25% smaller by weight (Fig C) (p<0.0001), indicating protection of the native flora. The findings demonstrate that commensal bacteria confer protection against fetal/placental ZIKV infection and transmission either through direct host defense or mediated through adaptive immunity.View Large Image Figure ViewerDownload Hi-res image Download (PPT)

Interleukin 11 blockade during mid to late gestation does not affect maternal blood pressure, pregnancy viability or subsequent fertility in mice.

Interleukin (IL)11 is a crucial regulator during the initiation of pregnancy in humans and mice. Elevated levels are detected in serum, placenta and decidua of women with pre-eclampsia. Elevated IL11 during placentation recapitulates pre-eclampsia in mice, although withdrawal rescues pre-eclampsia features, suggesting that IL11 could provide a novel therapeutic target. The aim of this study was to determine the safety profile of an IL11 antagonist ligated to polyethylene glycol (PEGIL11A) during pregnancy in mice. Blocking IL11 signalling during mid to late gestation pregnancy in mice did not affect pregnancy viability, or alter placental or fetal weight, or morphology. Importantly, decidual area remained unchanged. PEGIL11A did not affect maternal blood pressure, urinary protein or term pup weight. PEGIL11A administration to non-pregnant mice did not affect subsequent fertility; there was no difference in number of implantation sites, or placental or fetal weight between PEGIL11A and PEG-treated mice. These data show that blocking IL11Rα during placentation does not alter the placenta, decidua, fetus, maternal blood pressure or kidneys. These findings highlight the potential of IL11 signalling inhibition as a safe therapy to alleviate pre-eclampsia symptoms and demonstrate the potential for IL11 inhibition as a novel fertility-preserving therapy for women with cancer.

Continued Investigation Into 17-OHPC: Results From the Preclinical RUPP Rat Model of Preeclampsia.

Preeclampsia is characterized by elevated TNF-α (tumor necrosis factor-α), antiangiogenic factors, such as sFlt-1 (soluble vascular endothelial growth factor receptor 1), increased uterine artery resistance index, and decreased of NO during pregnancy. Previously we showed that 17-hydroxyprogesterone caproate (17-OHPC) administered into reduced uterine perfusion pressure (RUPP) rats on day 18 of gestation improved hypertension without improving pup weight. We hypothesized that earlier administration of 17-OHPC on day 15 of gestation could improve pathophysiology of preeclampsia and fetal outcomes in response to placental ischemia. Carotid catheters were inserted on day 18, and mean arterial blood pressure and samples were collected on day 19. Mean arterial blood pressure in normal pregnant rats was 102±2, 105±2 in normal pregnant+day 15 of gestation (GD15) 17-OHPC, 127±2 in RUPP and 112±1 mm Hg in RUPP+GD15 17-OHPC, P<0.05. Pup weight and litter size were improved from 1.9±0.05, 10.1±1.4 in RUPP to 2.1±0.07 g and 13.2±0.6 in RUPP+GD15 17-OHPC, P<0.05. Uterine artery resistance index was 0.8±0.03 in RUPP, which was decreased to 0.6±0.04 in RUPP+GD15 17-OHPC, P<0.05. Plasma TNF-α levels were 164±34 in RUPP and blunted to 29±9 pg/mL in RUPP+GD15 17-OHPC, P<0.05. Plasma nitrate-nitrite levels were 10.8±2.3 in RUPP rats and significantly increased to 25.5±5.2 µmol/L in RUPP+GD15 17-OHPC, P<0.05. sFlt-1 levels were 386±141 in RUPP rats, which were reduced to 110.2±11 in RUPP+17-OHPC, P<0.05. These data indicate that GD15 17-OHPC improves pathophysiology in RUPP rats, possibly via improving sFlt-1 reduced NO during pregnancy.

The effect of repeated light-dark shifts on uterine receptivity and early gestation in mice undergoing embryo transfer

ABSTRACTFemale shift workers are at increased risk for negative reproductive outcomes, and animal evidence suggests that manipulation of the light-dark cycle is detrimental to early gestation in female mice. Specifically, failure of implantation may be responsible for these findings. The objective of this study was to better delineate which reproductive processes are vulnerable to detrimental effects of maternal circadian disturbance. We exposed mice undergoing embryo transfer to repetitive phase advances of the photoperiod. Embryos were derived from donor sperm and eggs from mice living in normal light-dark conditions to isolate the effects of photoperiod disruption on uterine receptivity and early gestation. Twenty-eight mice receiving embryo transfer underwent an experimental light-dark condition (advance of lights on and lights off by 6 hours every 4 days). Twenty-eight mice remained in a normal light-dark condition. Animals lived in their assigned light-dark condition beginning 2 weeks prior to embryo transfer and ending the day of uterine necropsy (post-coitus day 14.5). Wilcoxon-Mann-Whitney test demonstrated no significant differences between control and experimental light-dark conditions in pups (Z=0.10, p=.92), resorptions (Z=0.20, p=.84), or implantations (Z=-0.34, p=.73). Pup and placental weights were similar between groups. In this investigation, uterine receptivity and maintenance of early gestation were preserved despite recurrent phase advances in photoperiod. This finding, in the context of the current literature, suggests that the negative effects of circadian disruption are mediated by reproductive processes upstream of implantation.

Characterization of mammary-specific disruptions for Tph1 and Lrp5 during murine lactation

Serotonin is a homeostatic regulator of the mammary gland during lactation. The contribution of mammary-derived serotonin to circulating serum serotonin concentrations was previously unknown. We have developed mice with mammary-specific disruptions of tryptophan hydroxylase 1 (Tph1) or low-density lipoprotein receptor-related protein 5 (Lrp5) that are induced during late pregnancy and lactation via use of the whey acidic protein (WAP)-Cre cre-lox system. Our objective was to characterize dams with a lactation- and mammary-specific disruption of Lrp5 (WAP-Cre × Lrp5FL/FL) or Tph1 (WAP-Cre × Tph1FL/FL). Milk yield and pup weights were recorded throughout lactation. Dams were euthanized on d10 postpartum and mammary glands and duodenal tissue were harvested. WAP-Cre × Lrp5FL/FL dams had elevated serotonin concentrations in both the mammary gland and circulation compared to controls. In contrast, WAP-Cre × Tph1FL/FL dams had decreased mammary gland and serum serotonin concentrations compared to controls. Alveolar morphology, milk yield, and pup weights were similar. Mammary-derived serotonin makes a significant contribution to circulating serotonin concentrations during lactation, with no effect on milk yield or alveolar morphology. These transgenic models can and should be confidently used in future lactation studies to further elucidate the contribution of serotonin to the maintenance of lactation.

Effect of high-fat diet on secreted milk transcriptome in midlactation mice.

High-fat diet (HFD) during lactation alters milk composition and is associated with development of metabolic diseases in the offspring. We hypothesized that HFD affects milk microRNA (miRNA) and mRNA content, which potentially impact offspring development. Our objective was to determine the effect of maternal HFD on secreted milk transcriptome. To meet this objective, 4 wk old female ICR mice were divided into two treatments: control diet containing 10% kcal fat and HFD containing 60% kcal fat. After 4 wk on CD or HFD, mice were bred while continuously fed the same diets. On postnatal day 2 (P2), litters were normalized to 10 pups, and half the pups in each litter were cross-fostered between treatments. Milk was collected from dams on P10 and P12. Total RNA was isolated from milk fat fraction of P10 samples and used for mRNA-Seq and small RNA-Seq. P12 milk was used to determine macronutrient composition. After 4 wk of prepregnancy feeding HFD mice weighed significantly more than did the control mice. Lactose and fat concentration were significantly ( P < 0.05) higher in milk of HFD dams. Pup weight was significantly greater ( P < 0.05) in groups suckled by HFD vs. control dams. There were 25 miRNA and over 1,500 mRNA differentially expressed (DE) in milk of HFD vs. control dams. DE mRNA and target genes of DE miRNA enriched categories that were primarily related to multicellular organismal development. Maternal HFD impacts mRNA and miRNA content of milk, if bioactive nucleic acids are absorbed by neonate differences may affect development.

Preconception exposure to dietary levels of genistein affects female reproductive outcomes

Genistein is a phytoestrogen found in soy and soy-based products. Previously, we found that genistein adversely affected estradiol levels and follicle growth in vitro. Proper hormone production and follicle growth are key regulators of normal fertility. Therefore, we hypothesized that genistein adversely affects female fertility and pregnancy outcomes. To test this hypothesis, we dosed sexually mature female CD-1 mice (35days) with 0, 300, 500, or 1000ppm genistein for 30, 60, 150, and 240days. At the end of the dosing periods, we measured mating rate, pregnancy rate, fertility rate, gestation time, parturition time, pup mortality, litter size, average pup weight, and estradiol and progesterone levels. We found that chronic, preconception exposure to genistein affects gestation time, parturition time, litter size, pup weight, and pup mortality. Additionally, genistein exposure for 240days appears to have a protective effect on fertility rate, but does not affect hormone levels in vivo.

Angiogenic imbalance and diminished matrix metalloproteinase-2 and -9 underlie regional decreases in uteroplacental vascularization and feto-placental growth in hypertensive pregnancy

Preeclampsia is a form of hypertension-in-pregnancy (HTN-Preg) with unclear mechanism. Generalized reduction of uterine perfusion pressure (RUPP) could be an initiating event leading to uteroplacental ischemia, angiogenic imbalance, and HTN-Preg. Additional regional differences in uteroplacental blood flow could further affect the pregnancy outcome and increase the risk of preeclampsia in twin or multiple pregnancy, but the mechanisms involved are unclear. To test the hypothesis that regional differences in angiogenic balance and matrix metalloproteinases (MMPs) underlie regional uteroplacental vascularization and feto-placental development, we compared fetal and placental growth, and placental and myoendometrial vascularization in the proximal, middle and distal regions of the uterus (in relation to the iliac bifurcation) in normal pregnant (Preg) and RUPP rats. Maternal blood pressure and plasma anti-angiogenic soluble fms-like tyrosine kinase-1 (sFlt-1)/placenta growth factor (PIGF) ratio were higher, and average placentae number, placenta weight, litter size, and pup weight were less in RUPP than Preg rats. The placenta and pup number and weight were reduced, while the number and diameter of placental and adjacent myoendometrial arteries, and MMP-2 and MMP-9 levels/activity were increased, and sFlt-1/PlGF ratio was decreased in distal vs proximal uterus of Preg rats. In RUPP rats, the placenta and pup number and weight, the number and diameter of placental and myoendometrial arteries, and MMP-2 and -9 levels/activity were decreased, and sFlt-1/PlGF ratio was increased in distal vs proximal uterus. Treatment with sFlt-1 or RUPP placenta extract decreased MMP-2 and MMP-9 in distal segments of Preg uterus, and treatment with PIGF or Preg placenta extract restored MMP levels in distal segments of RUPP uterus. Thus, in addition to the general reduction in placental and fetal growth during uteroplacental ischemia, localized angiogenic imbalance and diminished MMP-2 and MMP-9 could cause further decrease in placental and myoendometrial vascularization and placental and fetal growth in distal vs proximal uterus of HTN-Preg rats. Regional differences in uteroplacental perfusion, angiogenic balance and MMPs could be a factor in the incidence of preeclampsia in multiple pregnancy.

Supplementing dams with both arachidonic and docosahexaenoic acid has beneficial effects on growth and immune development

Omega-3 long-chain polyunsaturated fatty acids (LCPUFAS) modulate immune cells in vitro and in vivo. This study investigated the effects of enriching the maternal diet with the n-6 and n-3 LCPUFAs, arachidonic (20:4n-6, 0.6%wt ARA) and docosahexaenoic acid (22:6n-3, 0.32%wt DHA), or 1:1 and 2:1 ratios (ARA: DHA) on total lipids in milk, total lipids, and immunophenotypes in plasma, lymph nodes, and spleen from isolated immune cells from 28d old pups. From day 15 of gestation to day 3 pp, Sprague-Dawley dams were fed a commercial chow. On day 3 pp litters were culled and pups (4 males and 2 females) randomly cross-fostered to dams who were randomized to one of the 5 experimental diets resulting in 20 male and 10 female pups/diet group. Dams fed ARA or ARA: DHA had 28–36% more 20:4n-6 in milk and feeding DHA or ARA: DHA doubled 22:6n-3 in milk lipids (P<0.05). Feeding 1:1 or 2:1 ARA: DHA resulted in greater pup weight at weaning (P<0.05). Compared to the control pups, ARA + DHA fed pups had a lower proportion of splenic CD45RA+ lymphocytes. In summary, postpartum supplementation with a combination of ARA + DHA, compared to ARA or DHA alone, resulted in a higher content of ARA and DHA in dam's milk and tissues and had positive effects on growth, accompanied by evidence of progression toward a mature immune phenotype, and suggests a need for ARA when DHA is supplemented in the early diet. Additional investigations are needed of ARA immunomodulation to better understand and estimate nutritional requirements for LCPUFAs during early development.

The adverse effects of perinatal exposure to nonylphenol on carbohydrate metabolism in male offspring rats

Background: We sought to investigate the adverse effects of perinatal exposure to nonylphenol (NP) on carbohydrate metabolism of male offspring rats. Methods: Thirty-two healthy pregnant Sprague Dawley rats were randomly divided into four groups, control normal diet group (C), NP normal diet group (NPN), control high-energy diet group (CH), and NP high-energy diet group (NPH). Both of the control groups were received a gavage of corn oil and the NP-groups were received NP (200 mg/kg/day) from gestational days 6 to post-natal day (PND) 21. The concentrations of NP in pancreatic tissues were measured by high-performance liquid chromatography (HPLC). The key genes of glucose metabolism expression were detected by reverse transcription-polymerase chain reaction (RT-PCR). The pancreatic tissues were stained with hematoxylin/eosin (HE). Results: On PND 1, the body weights of male pups in the NPN and NPH groups were lower than those of the CH group (p = 0.012 and 0.001, respectively). On PND 30, the body weight of male pups from the NPH group was elevated compared with the C group (p = 0.019), while the body weights of male pups in the NPN and NPH groups were elevated compared to the CH group (p = 0.034 and 0.004, respectively). On PND 60, the body weights of NPN and NPH pups were higher than those in the C (p < 0.001) and CH groups (p < 0.001). The levels of fasting blood glucose (FBG) were increased significantly in the animals treated with NP compared to control animals (F = 29.14, p < 0.001). The FBG levels in the treatment groups are ranked as follows: NPH > NPN > CH > C (p < 0.05). The concentrations of NP in pancreas tissues in both the NPN (2045.0 ± 130.1 μg/L) and NPH groups (2038.0 ± 104.2 μg/L) were higher than those in the C (499.5 ± 27.4 μg/L) and CH groups (494.2 ± 22.4 μg/L; p < 0.05). Morphological examination of tissues from rats exposed to NP shown that the NP-treated groups appeared to have a higher degree of inflammatory injury, edema, and focal necrotic cells in the pancreatic tissues. Compared with C group, expression of glucokinase (GCK) was down-regulated, while Uncoupling protein-2 (UCP-2) was up-regulated in the NP-treated groups (FGCK = 218.89, p < 0.001; FUCP-2 = 18.82, p < 0.001). Conclusions: Prenatal exposure to NP could induce glucose metabolism disorder in male F1 rats, which may be due to the fact that NP induces abnormal expression patterns of GCK and UCP-2.