Introduction and Objective: A high fat diet (HF) during pregnancy (P) and lactation (L) induces kidney disease in offspring. In diabetic kidney disease renal GLUT2 is increased. Mice with a genetic heterozygous deletion of GLUT4 (G4+/-), a model of insulin resistance, develop renal disease if exposed to HF during P/L. This study investigates the effect of a HF during P/L on the metabolic phenotype and expression of glucose homeostasis genes in kidney in male G4+/- and wild type (WT) offspring. Methods: Female CD1 mice were fed either a HF or control diet (C) prior to mating with G4+/- males and throughout P/L. After weaning male offspring were fed a C diet. Body weight (BW), kidney weights and blood glucose were measured. Mice were sacrificed at 5 or 13 wks, respectively, and renal GLUT1, 2 and 8 mRNA was measured by qRT-PCR (n=4-6). Results: BW of G4+/- mice on HF during P/L was lower by 11.7% at 5 wks and by 17.4% at 13 wks compared to G4+/- mice on C whereas BW of WT mice on HF during P/L was lower by 18.5% compared to WT on C during P/L at 13wks only (P<0.05). Kidney weight was reduced in both G4+/- on C (10.5%) and on HF (11.2%) during P/L compared to WT controls at 5 wks but was increased at 13 wks in G4+/- (1.15-fold) and in WT (1.22-fold) on HF during P/L compared to their genotype on C during P/L (P<0.05). No differences in blood glucose were observed. For GLUT2 gene expression, an increase in G4+/- mice on C during P/L (1.69- fold and 1.52- fold compared to WT on C and G4+/- on HF during P/L, respectively) was observed at 5 wks whereas at 13 wks an increase was observed for WT on HF during P/L compared to G4+/- (1.34-fold) and to WT (1.55-fold) on C during P/L (P<0.05). A decrease in GLUT8 gene expression by 47.3% compared to WT on C during P/L (P=0.001) was observed at 5 wks. No differences in GLUT1 gene expression were observed. Conclusion: A HF diet during P/L alters renal morphology and expression of GLUT2 which both indicate renal dysfunction, already early in post-weaning life in the absence of alterations in blood glucose levels. M. Kruse: None. Y. Seki: None. X. Du: None. M.J. Charron: None.

The purpose of this study was to examine the deposition of advanced glycation end products (AGEs) and their receptors, RAGE, in ovarian follicles during folliculogenesis in mice fed high (H-AGE) or low (L-AGE) AGE diets and following superovulation with gonadotropins. We hypothesize that H-AGE diet is associated with increased AGE deposition and RAGE expression in various stages of ovarian follicular development, and superovulation with gonadotropins may alter these changes. C57BL/6J mice were fed low L-AGE (n = 10) or H-AGE (n = 10) diet for 12 weeks. In each group, half of each cohort (n = 5) were sacrificed at the end of 12 weeks while the other half (n = 5) were superovulated prior to sacrifice. Immunofluorescence staining of ovarian sections was used to determine AGE deposition and RAGE expression in ovarian follicles in a semi-quantitative manner. In all mice, AGE deposition and RAGE expression were observed in granulosa but not theca cells. In all mice, AGE deposition intensity increased as the follicles progressed through developmental stages from primordial to primary to secondary to prenatral/antral but then significantly dropped in the corpus luteum stage. RAGE staining was highly expressed equally in all stages of pre-ovulatory follicles but then significantly dropped in the corpus luteum post-ovulatory stage. Compared to mice on L-AGE diet, mice on H-AGE mice had significantly lower AGE deposition in their primordial follicles and lower RAGE intensity in their antral follicles. Following superovulation, mice in both groups had significantly lower AGE deposition and significantly lower RAGE expression but the drop in AGE deposition following superovulation was more pronounced in the H-AGE diet group compared to the L-AGE diet group in both pre-ovulatory and post-ovulatory follicles. Ovarian AGE deposition and RAGE expression changes differently during the natural follicular development. Opposite to expectations, the intake of diet rich in AGEs caused lower expression of the proinflammatroy RAGE, an effect that was more pronounced after gonadotropin exposure.

PROBLEM:Exposures during the perinatal period, a phase of rapid development and growth, may have a profound and sustained effect on metabolic disturbances later in life. The pro-inflammatory advanced glycation end products (AGEs) are widely consumed in the Western diet. The purpose of this study was to determine whether perinatal exposure to these dietary AGEs alters metabolic parameters, in particular adiposity and glucose hemostasis, in male mice offspring. METHODS:Seven-week-old female CD1 mice were placed before mating and then throughout pregnancy and lactation on either a low AGE (L-AGE; n = 13) or high AGE (H-AGE; n = 13) diet. All offspring in both groups were weaned postnatal day 21 onto normal diet and studied through to 21 weeks of age. The offspring were counted and weighed weekly, starting at birth until 21 weeks of age, to assess the growth curve. At the time of sacrifice, Echo MRI was performed to measure adiposity and to record liver, white epididymal adipose tissue (WAT), and inguinal fat weights. Serum levels of leptin as well as insulin and glucose tolerance tests (ITT and GTT) were compared. RESULTS:The Body weight at birth of offspring of dams that were on H-AGE diet was significantly lower compared to the body weight of offspring of dams that were on L-AGE diet. Echo MRI data showed that the offspring of dams that were H-AGE diet had significantly lower fat mass, lower epididymal WAT fat weight, and lower inguinal fat weight but higher lean body mass and similar liver weight. They also had significantly higher glucose levels during GTT and ITT, as well as significantly lower serum leptin levels compared to the offspring of dams that were on the L-AGE diet. CONCLUSIONS:These results indicate that perinatal exposure to a maternal diet elevated in AGEs causes deficits in perinatal growth and impairment in glucose hemostasis in male mice. These findings suggest that AGEs may represent an important new class of mediators of adiposity and the metabolic syndrome.

The glucagon receptor (GCGR) in the kidney is expressed in nephron tubules. In humans and animal models with chronic kidney disease, renal GCGR expression is reduced. However, the role of kidney GCGR in normal renal function and in disease development has not been addressed. Here, we examined its role by analyzing mice with constitutive or conditional kidney-specific loss of the Gcgr. Adult renal Gcgr knockout mice exhibit metabolic dysregulation and a functional impairment of the kidneys. These mice exhibit hyperaminoacidemia associated with reduced kidney glucose output, oxidative stress, enhanced inflammasome activity, and excess lipid accumulation in the kidney. Upon a lipid challenge, they display maladaptive responses with acute hypertriglyceridemia and chronic proinflammatory and profibrotic activation. In aged mice, kidney Gcgr ablation elicits widespread renal deposition of collagen and fibronectin, indicative of fibrosis. Taken together, our findings demonstrate an essential role of the renal GCGR in normal kidney metabolic and homeostatic functions. Importantly, mice deficient for kidney Gcgr recapitulate some of the key pathophysiological features of chronic kidney disease.

Obesity induced by a high fat (HF) diet during pregnancy (P) and lactation (L) shows a sexual dimorphic response in offspring. Studies reported a metabolic benefit for female compared to male offspring. The mechanisms that determine sexual dimorphic responses are still poorly understood. This study investigates the metabolic phenotype, adipose tissue distribution and hepatic lipid metabolism in female offspring exposed to a HF diet during P/L and early adulthood. CD1 female offspring mice were exposed to either a control (C) or a HF diet during P/L and fed a regular chow after weaning for 23 wks and then re-exposed to either the C or HF diet for 19 wks. This resulted into mice exposed to the C diet during P/L and re-exposed to either the C (C-C) or HF (C-H) diet, and mice exposed to the HF diet during P/L and re-exposed to either the C (H-C) or HF (H-H) diet (n=3-5). Body weight (BW), body composition, food intake (FI), fasted blood glucose and insulin levels, intra peritoneal glucose (ipGTT) and insulin (ipITT) tolerance were measured. Mice were sacrificed at 45 wks and qRT-PCR of stearoyl-CoA-desaturase 1 (SCD-1) as a marker of lipogenesis and of caveolin-1 (CAV1) as a protective marker of hepatic steatosis was performed in liver. BW, adiposity and ipGTT and ipITT were not different between C-H and H-H. However adiposity was increased 3.0-fold in C-H compared to C-C and 2.0-fold in H-H compared to H-C, respectively (both p<0.05). FI and fasted glucose and insulin were not different between groups. Gene expression of SCD-1 was downregulated by 69% in C-H compared to C-C and by 81% in H-H compared to H-C, respectively, (both p<0.05) in liver. In contrast, hepatic CAV1 gene expression was increased 1.93-fold in H-H compared to H-C (p<0.05). These data show that female mice are protect from the deleterious programming effects of a HF diet. Moreover, mice exposed to a HF diet during P/L exhibit protective mechanisms to prevent hepatic steatosis when re-exposed to HF diet in early adulthood. Disclosure M. Kruse: None. Y. Seki: None. X. Du: None. M.J. Charron: None.

Neonatal and early-life gut microbiome changes are associated with altered cardiometabolic and immune development. In this study, we explored Cesarean delivery effects on the gut microbiome in our high-risk, under-resourced Bronx, NY population. Fecal samples from the Bronx MomBa Health Study (Bronx MomBa Health Study) were categorized by delivery mode (vaginal/Cesarean) and analyzed via 16 S rRNA gene sequencing at four timepoints over the first two years of life. Bacteroidota organisms, which have been linked to decreased risk for obesity and type 2 diabetes, were relatively reduced by Cesarean delivery, while Firmicutes organisms were increased. Organisms belonging to the Enterococcus genus, which have been tied to aberrant immune cell development, were relatively increased in the Cesarean delivery microbiomes. Due to their far-reaching impact on cardiometabolic and immune functions, Cesarean deliveries in high-risk patient populations should be carefully considered.

Babies born to severe acute respiratory syndrome corona virus-2 (SARS-CoV-2)-infected mothers are at greater risk for perinatal morbidity and more likely to receive a neurodevelopmental diagnosis in the first year of life. However, the effect of maternal infection on placental function and neonatal outcomes varies depending upon the patient population. We set out to test our hypothesis that maternal SARS-CoV-2 infection in our underserved, socioeconomically disadvantaged, mostly unvaccinated, predominantly African American and Latina population in the Bronx, NY would have effects evident at birth. Under IRB approval, 56 SARS-CoV-2-positive patients infected during the "first wave" of the pandemic with alpha and beta strains of the virus, 48 patients infected during the "second wave" of the pandemic with delta and omicron strains and 61 negative third-trimester high-risk patients were randomly selected from Montefiore Medical Center (MMC), Bronx, NY. In addition, two positive cases from Yale New Haven Hospital, CT were included as controls. All 104 placentas delivered by SARS-CoV-2-positive mothers were uninfected by the virus, based on immunohistochemistry, in situ hybridization, and qPCR analysis. However, placental villous infarcts were significantly increased in first-wave cases compared to second-wave cases or negative controls. Significantly lower Apgar scores at 1 min and 5 min were observed in neonates born to infected mothers with severe symptoms. These findings suggest that even without entering the placenta, SARS-CoV-2 can affect various systemic pathways, culminating in altered placental development and function, which may adversely affect the fetus, especially in a high-risk patient population such as ours. These results underline the importance of vaccination among pregnant women, particularly in low-resource areas.

Intrauterine growth restriction (IUGR) is associated with increased risk of cardiometabolic disease later in life and has been shown to affect female and male offspring differently, but the mechanisms remain unclear. The purpose of this study was to identify proteomic differences and metabolic risk markers in IUGR male and female neonates when compared to appropriate for gestational age (AGA) babies that will provide a better understanding of IUGR pathogenesis and its associated risks. Our results revealed alterations in IUGR cord plasma proteomes with most of the differentially abundant proteins implicated in peroxisome pathways. This effect was evident in females but not in males. Furthermore, we observed that catalase activity, a peroxisomal enzyme, was significantly increased in females (p < 0.05) but unchanged in males. Finally, we identified risk proteins associated with obesity, type-2 diabetes, and glucose intolerance such as EGF containing fibulin extracellular matrix protein 1 (EFEMP1), proprotein convertase subtilisin/kexin type 9 (PCSK9) and transforming growth factor beta receptor 3 (TGFBR3) proteins unique to females while coagulation factor IX (C9) and retinol binding protein 4 (RBP4) are unique in males. In conclusion, IUGR may display sexual dimorphism which may be associated with differences in lifelong risk for cardiometabolic disease between males and females.

IntroductionThe effects of psychological distress/resilience on parent-child engagement (e.g., family dinners, reading) during the COVID-19 pandemic have not been well studied. Among very young children from underrepresented backgrounds enrolled in the ongoing longitudinal Bronx Mother Baby Health Study of healthy term infants, we (1) examined associations between exposures to COVID-19-related events, demographic factors and parental psychological distress and resilience; and (2) correlated these factors with parent-child engagement activities.MethodsBetween June 2020-August 2021, parents of 105 Bronx Mother Baby Health Study participants aged birth-25 months completed questionnaires related to exposures to COVID-19-related events, frequency of positive parent-child engagement activities, food and housing insecurity, and parental psychological distress and resilience. Families were also asked open ended questions about the pandemic's impact.Results29.8% and 47.6% of parents reported food and housing insecurity, respectively. Greater exposures to COVID-19-related events were associated with increased parental psychological distress. Positive parent-child interactions were associated with demographic factors and higher levels of maternal education, but not with exposures to COVID-19-related events.DiscussionThis study adds to a growing body of literature on the negative impacts of COVID-19 exposures and psychosocial stressors on families during the pandemic, supporting the need for enhanced mental health resources and social supports for families.

Objective: To examine, following perinatal exposure to a diet high in advanced glycation end products (AGEs), whether the use of standard AGE-free mouse chow during the postweaning period alters metabolism and reproduction differently than exposure to a diet low in AGEs. Design: Experimental animal study. Setting: University-based research laboratory. Animal(s): Female CD1 mice. Intervention(s): Seven-week-old mice were placed on a diet either low or high in AGEs perinatally, before mating and then during pregnancy and lactation. All offspring were weaned onto an AGE-free normal chow. Main Outcome Measure(s): Growth curve, liver and abdominal fat weight, insulin and glucose tolerance tests, vaginal opening, estrous cyclicity, and serum levels of antimullerian hormone, leptin, and adiponectin were assessed. Ovarian histologic examination for follicular count and gene expression was also performed. Result(s): Compared with the mice exposed to a diet low in AGEs, the mice exposed to a diet high in AGEs showed lower body weight in pups, lower liver weight, delayed vaginal opening, higher serum antimullerian hormone levels, lower primordial and secondary follicle pools, and higher ovarian Fshr messenger RNA levels. Conclusion(s): Following weaning, perinatal AGEs can target puberty onset and folliculogenesis differently to standard mouse chow. (Fertil Steril Sci (R) 2022;3:95-105. (c) 2021 by American Society for Reproductive Medicine.)

Epidemiologic studies have shown an association between an adverse intrauterine environment (eg, exposure to malnutrition) and an increased risk of developing cardiometabolic disease in adulthood. These studies laid the foundation for the developmental origins of health and disease hypothesis, which states that limited nutrient supply to the fetus results in physiologic and metabolic adaptations that favor survival but result in unfavorable consequences in the offspring if there is excess nutrition after birth. This discrepancy in the pre- and postnatal milieus, perceived as stress by the offspring, may confer an increased risk of developing cardiometabolic disease later in life. Thus, early life exposures result in programming or changes in cellular memory that have effects on health throughout the life course. One of the mechanisms by which programming occurs is via epigenetic modifications of genes, processes that result in functionally relevant changes in genes (ie, gene expression) without an alteration in the genotype. In this review, we will describe how fetal exposures, including under- and overnutrition, affect neonatal and childhood growth and the future risk for cardiometabolic disease.

Abstract. Babies born to severe acute respiratory syndrome corona virus-2 (SARS-CoV-2) infected mothers are at greater risk for perinatal morbidity and more likely to receive a neurodevelopmental diagnosis in the first year of life. However, the effect of maternal infection on placental function and neonatal outcomes varies depending upon the patient population. We set out to test our hypothesis that maternal SARS-CoV-2 infection in our underserved, socioeconomically disadvantaged, predominantly African American and Latina population in the Bronx, NY would have effects evident at birth. Fifty-five SARS-CoV-2 positive and 61 negative third trimester patients were randomly selected from Montefiore Medical Center (MMC), Bronx, NY. In addition, two positive cases from Yale New Haven Hospital, CT were included as controls. All 55 placentas delivered by SARS-CoV-2 positive mothers were uninfected by the virus, based on immunohistochemistry, in-situ hybridization, and qPCR analysis. However, placental villous infarcts, mild preeclampsia, shortened gestational periods and lower Apgar scores were observed in the infected cases. These findings suggest that even without entering the placenta, SARS-CoV-2 can affect various systemic pathways culminating in altered placental development and function, which may adversely affect the fetus, especially in a high-risk patient population such as ours. These results underline the importance of vaccination among pregnant women, particularly in low resource areas.

An adverse maternal in utero environment can program offspring for increased risk for metabolic disease. The aim of this study was to determine whether N-acetylcysteine (NAC), an anti-inflammatory antioxidant, attenuates programmed susceptibility to obesity and insulin resistance in high fat diet (HFD) offspring. CD1 female mice were acutely fed a standard breeding chow or HFD. NAC was added to the drinking water (1g/kg) of the treatment cohorts from embryonic day 0.5 (e0.5) until the end of lactation. NAC treatment normalized HFD-induced maternal weight gain and oxidative stress, improved the maternal lipidome and prevented maternal leptin resistance. These favorable changes in the in utero environment normalized postnatal growth, decreased white adipose tissue (WAT) and hepatic fat, improved glucose and insulin tolerance and antioxidant capacity, reduced leptin and insulin and increased adiponectin in HFD offspring. The lifelong metabolic improvements in the offspring were accompanied by reductions in pro-inflammatory gene expression in liver and WAT and increased thermogenic gene expression in brown adipose tissue (BAT). These results, for the first time, provide a mechanistic rationale for how NAC can prevent the onset of metabolic disease in the offspring of mothers who consume a typical Western HFDs.

Maternal nutrition and the intrauterine environment are important in determining susceptibility to reproductive and metabolic disturbances. Advanced glycation end products (AGEs) are widely consumed in Western diet. The purpose of this study was to determine whether perinatal exposure to a high levels of dietary AGEs affect metabolic and reproductive parameters in female mice offspring. Female CD1 mice, 7 weeks old, were placed on either a diet low (L-AGE) or high (H-AGE) in AGEs before mating and then during pregnancy and lactation. All offspring were weaned onto the L-AGE diet and studied through to 16 weeks of age; they were counted and weighed at birth and then every week for a total of 11 weeks. Vaginal opening, litter size, growth curve, liver and abdominal fat weights, serum levels of anti-Mullerian hormone, leptin and adiponectin, as well as insulin and glucose tolerance tests were compared. Ovaries were harvested for follicular count and gene expression by real-time polymerase chain reaction. Compared to perinatal exposure to the L-AGE diet, perinatal exposure to the H-AGE diet caused lower body weight at birth, and adult offspring exhibited delayed growth, lower serum leptin and adiponectin levels, delayed vaginal opening, irregular oestrous cyclicity, arrested follicular development and significant alterations in the expression of genes involved in folliculogenesis (Amh and Amhr2) and steroidogenesis (Cyp19a1). These results indicate that perinatal exposure to a diet elevated in AGEs causes deficits in perinatal growth, pubertal onset, and reproductive organ development in female mice. Whether these findings translate to humans remains to be determined in future studies.

Obesity, known to cause a systemic elevation in monocyte chemotactic protein-1 (MCP-1), adversely affects normal ovarian function. The aim of this study was to determine whether MCP-1 plays a role in ovarian dysfunction that is related to obesity induced by high-fat (HF) diet intake. Wild type (WT) C57BL/6J mice were fed either normal chow (NC) (Group 1, control group) or HF diet (Group 2). To assess whether MCP-1 is involved in HF-diet-induced ovarian dysfunction, MCP-1 knock-out mice were fed HF diet (Group 3). Body weight, body fat composition, number of oocytes collected following ovarian superovulation with gonadotropins, ovarian macrophage markers and expression of genes important in folliculogenesis and steroidogenesis were quantified in the 3 groups of animals. Animals in Group 2 gained significant body weight and body mass, produced the fewest number of oocytes following superovulation, and had significant alterations in ovarian genes involved in folliculogenesis and steroidogenesis as well as genes involved in inflammation. Although animals in Group 3 had the highest body weight and body fat composition, they produced similar number of oocytes compared to animals in Group 1 but had different ovarian gene expression compared to Group 2. These findings suggest that MCP-1 gene knockout could reverse some of the adverse effects of obesity induced by HF diet intake. Future studies assessing ovarian histology in MCP-1 knock out mouse model will confirm our findings. MCP-1 inhibition could represent a future therapeutic target to protect ovarian health from the adverse effects of HF diet ingestion.

Nutrition is an important source of exogenous AGEs and thermally processed foods present in western-style diets contain a large amount of these pro-inflammatory AGEs. Additionally, the intake of dietary AGEs could upregulate ovarian gene expression of inflammatory macrophage markers. The objective of this study was to investigate the effect of diet rich in AGEs on estrous cyclicity and ovarian function in a mouse model. Six-week old C57BL/6 J female mice were randomly subjected to either a diet low in AGEs (L-AGE) or a diet high in AGEs (H-AGE) for a total of 13 weeks. Experiments performed included daily vaginal smears to assess estrous cyclicity, ovarian superovulation with gonadotropins to assess the number of oocytes released, whole ovarian tissue mRNA quantification by RT-PCR to quantify genes involved in folliculogenesis, steroidogenesis, and macrophage markers, and ovarian morphology for follicle count. Outcome measures included estrous cyclicity, number of oocytes following superovulation, expression of genes involved in folliculogenesis, steroidogenesis, and macrophage infiltration as well as the number of primordial, primary, secondary, antral follicles and corpora lutea. Compared to mice on L-AGE diet, mice on H-AGE spent significantly longer time in the diestrus phase, had similar number of oocytes released following ovarian superovulation, and showed significant alterations in genes involved in steroidogenesis (increase in Star mRNA expression levels) and folliculogenesis (increase in Gdf-9 and Fshr mRNA expression levels). Mouse macrophage marker F4/80 mRNA expression was upregulated in mice on H-AGE diet compared to mice on L-AGE diet. Finally, mice on H-AGE diet had significantly fewer corpora lutea in their ovaries. These results indicate that the ingestion of high amounts of dietary AGEs could disrupt folliculogenesis and steroidogenesis that might lead to abnormal estrous cyclicity. Intake of dietary AGEs could also upregulate ovarian gene expression of inflammatory macrophage markers.

Various endocrine factors contribute to cold-induced white adipose tissue (WAT) browning, but glucagon has largely been ignored. The purpose of the current investigation was to determine if glucagon was required for the effects of cold on WAT browning. Utilizing whole-body glucagon receptor knockout (Gcgr(-/-)) mice and their wild-type (WT) littermate controls, we examined the response of inguinal WAT (iWAT) and interscapular brown adipose tissue (BAT) to an acute (48 h) cold stress or challenge with the beta 3-adrenergic agonist CL316,243. The effects of glucagon alone on the induction of thermogenic genes in adipose tissue from C57BL6/J mice were also examined. Gcgr(-/-) mice displayed modest increases in indices of browning at room temperature while displaying a blunted induction of Ucp1, Cidea, and Ffg21 mRNA expression in iWAT following cold exposure. Similarly, cold induced increases in mitochondrial DNA copy number, and the protein content of mitochondrial respiratory chain complexes, UCP1, and PGC1 were attenuated in iWAT from Gcgr(-/-) mice. In BAT, the induction of thermogenic markers following cold exposure was reduced, but the effect was less pronounced than in iWAT. Glucagon treatment increased the expression of thermogenic genes in both iWAT and BAT of C57BL6/J mice. In response to CL316,243, circulating fatty acids, glycerol, and the phosphorylation of hormone-sensitive lipase were attenuated in iWAT of Gcgr(-/-) mice. We provide evidence that glucagon is sufficient for the induction of thermogenic genes in iWAT, and the absence of intact glucagon signaling blunts the cold-induced browning of WAT, possibly due, in part, to impaired adrenergic signaling.