Abstracts
Cancer Diseases (CD)
CD1. Alpha-zeranol induces mammary gland proliferation in an ACI rat model and increases stem cell formation in ER+ breast cancer.
C. Winz, K. Chang Lee, B. Liu, Dr. P. Furmanski, Dr. N. Suh
Rutgers University Department of Chemical Biology, Rutgers University Ernest Mario School of Pharmacy
Estrogenic endocrine disrupting compounds (EDCs) found in plastics, drinking water, and food, pose a threat to human health. In the US, the proportion of breast cancer diagnoses of ductal origin have increased dramatically over the past 50 years, with no identified cause. Ductal breast cells have proliferative capacity in response to endocrine signaling, due to their role in lactation. The increase in ductal carcinomas of the breast has led to concerns that exposure to estrogenic EDCs may contribute to the incidence and progression of breast cancer. EDCs share chemical similarity with the endogenous hormone estrogen, and are thus able to activate estrogen receptor related signaling pathways that lead to abnormal cell proliferation, differentiation, and migration. Our laboratory has demonstrated that chronic estrogen exposure in the ACI rat model results in mammary proliferation at early timepoints and tumorigenesis at later timepoints. Our laboratory and others have additionally shown that the endogenous estrogen β-estradiol (E2) increases the population of cancer stem cells (CSCs) in ER+ breast cancer. Researchers have identified that CSCs are the only cancer cell type that can repopulate tumors in vivo. Clinically, increased numbers of CSCs have been associated with metastasis and recurrence in breast cancer patients. Since estrogenic EDCs share chemical similarity with E2, they may also induce cancer stemness in ER+ breast cancer, resulting in poorer disease outcomes. However, the molecular mechanisms behind EDC-induced cancer stemness have not been elucidated. We hypothesized that, due to its chemical similarity, alpha zeranol (aZAL), a well-known mycoestrogenic grain contaminant, may increase proliferation of the mammary gland in the ACI rat model and activate cancer stem cells in ER+ breast cancer cell lines. In ACI rats, aZAL implantation at a 6 mg dose induced proliferative changes in the mammary tissue at a five-day timepoint. Mammary gland expression of the estrogen receptor target progesterone receptor (PGR) and the proliferative cell marker PCNA also increased at the 5 day timepoint, as evidenced by upregulation in mRNA expression and increased immunohistochemical staining. Additionally, we employed a sphere-formation assay to assess stem cell induction in the ER+ MCF-7 cell line. We found that pretreatment with aZAL in monolayer conditions resulted in significant increases in sphere formation and size, indicating an increase in stem cell quantity and function. Taken together, these results demonstrate that aZAL is a concerning endocrine disruptor that may have distinct effects during the initiation phase and progression of mammary carcinogenesis.
CD2. Relationship between F-Box Protein 45 (FBXO45) and the tumor suppressor Prostate apoptosis response protein 4 (Par-4) in cancer: a proof of concept study
N. Sengottuvelan, A. Sahasrabuddhe, R. Wu, K.S.J. Elenitoba-Johnson, M. So-Young Lim
High School Student (Junior Year) Hillsborough High School, Hillsborough, NJ 08844, Assistant Lab Member, Memorial Sloan Kettering Cancer Center, New York, NY 10065, Senior Research Scientist, Memorial Sloan Kettering Cancer Center, New York, NY 10065, Chair, Department of Pathology and Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, Director, Lymphoma Translational Research Program, Memorial Sloan Kettering Cancer Center, New York, NY 10065.
Ubiquitination is a post‐translational modification that regulates several critical cellular processes, including cell homeostasis, cell cycle, and neoplastic transformation. Ubiquitination is orchestrated by the ubiquitin-proteasome system (UPS), which constitutes a cascade of enzymes, namely ubiquitin-activating enzyme E1, ubiquitin-conjugating enzyme E2, and ubiquitin ligase E3. The E3 ligases are substrate-recognizing subunits of UPS that recruit substrates for ubiquitination. Cullin-RING E3 ligases are the largest class of these ligases and constitute multi-subunit enzymes that transfer ubiquitin onto protein substrates. F-Box Protein 45 (FBXO45), a variable component of the Cullin-RING E3 ubiquitin ligase complex that recognizes and degrades Prostate apoptosis response protein 4 (Par-4) that selectively induces apoptosis in cancer cells. This proof of concept (POC) study aims to determine whether there is any relationship between FBXO45 and Par-4 expression. Using HEK 293T cells, we evaluated the effects of Fbxo45 overexpression and knockdown on the expression of Par-4 at gene and protein levels. Our Western blot, Immunofluorescence, and qRT-PCR data show that Fbxo45 overexpression decreased Par-4 and knockdown increased Par-4 expression. Our results support the hypothesis that when Fbxo45 is silenced, Par 4 signals increase and might promote apoptosis-mediated cellular events.
CD3. PRENATAL ALCOHOL EXPOSURE (PAE) INREASED DEVELOPMENTAL PLURIPOTENCY ASSOCIATED FACTOR 4 EXPRESSION IN PITUITARY TUMOR THROUGH EPIGENETIC MODIFICATION AT THE PROMOTER REGION
U. Das, S. Chaudhary, O. Gangisetty, D. K. Sarkar
Endocrine Research Facilities, Department of Animal Science, Rutgers University, New Jersey 08901
We have recently found an oncogenic role of developmental pluripotency-associated 4 (Dppa4) gene, known to be involved in stem cell reprograming, in the pituitary of fetal alcohol exposed female rats. However, the mechanism by which ethanol increased the Dppa4 expression in tumor cells is still elusive. The epigenetic processes, including DNA methylation, histone modifications are thought to influence the gene expression chiefly at the transcription level. Therefore, we intrigued to find out PAE mediated epigenetic modification at promoter region of Dppa4 gene. To achieve our goal, we performed RNA seq, qPCR array, ChIP assay and finally we applied blocker in in-vivo system to confirm the results obtained in cell lines. We prepared pituitary cell lines from fetal alcohol exposed rat pituitary tumors (AF cells) and control rat pituitary tumors (AD cells). The key epigenetic regulators were identified from RNA seq data and qPCR array was performed to check the expression of these regulators. The enzymes associated with H3K4me3, H3K9ac, H3K36me3 and DNA hypomethylation were overexpressed in AF cells compared to AD while enzymes involved in H3K27me3, and DNA hypermethylation were downregulated in AF cells compared to AD. It is worthy to mention here that the genes linked with H3K4me3, H3K36me3 and DNA methylation showed the maximum fold changes among the other genes. The ChIP assay was performed to confirm the changes in H3K4me3, H3K36me3 and H3K27me3 under estrogen challenged condition after PAE. A significant fold enrichment in H3K4me3 with a lower H3K27me3 was seen at the Dppa4 promoter region. Though H3K36me3 showed an insignificant change in promoter region of Dppa4, the level was significantly high in gene body. Additionally, our results confirmed that PAE (with out estrogen challenge) modulated the epigenetic regulators to cause H3K4me3 at Dppa4 promoter. Finally, we used MM-101 blocker specific to the MLL1 catalytic subunit of H3K4 methyltransferase enzyme in in-vivo model. AF rat treated with blocker showed decrease level of H3K4me3 at the promoter with the decreased expression of DPPA4 in immunoblot data. Taken together, it can be concluded that PAE epigenetically modifying the Dppa4 promoter to increase its expression followed by aggressive tumor growth.
CD4. FETAL ALCOHOL EXPOSURE PROMOTES THE DEVELOPMENT OF AGGRESSIVE PROLACTIN-SECRETING PITUITARY TUMORS IN MALE RATS
S. Chaudhary, D. Sarkar
Rutgers Endocrine Research Program, Department of Animal Sciences, Rutgers University, New Brunswick, NJ 08901, USA, Rutgers University
We have recently shown that estrogen-induced prolactin secreting pituitary tumors are aggressive in fetal alcohol-exposed female rats. In this study, we studied whether estrogen-induced prolactin secreting pituitary tumors are also aggressive in fetal alcohol exposed male rats. To test this, pregnant Fischer 344 rats were fed between gestational days 7 and 21 with a liquid diet containing ethanol 6.7% v/v (AF), pair-fed with isocaloric liquid diet (PF) or fed with ad libitum rat chow (AD). Alcohol feeding using liquid diet in Fisher rat dams elevated the blood alcohol concentration to 120-150 mg/dl at 2h after the last feeding. At 60 days of age, male offspring rats were orchiectomized and received subcutaneous estradiol implants. These rats were sacrificed at 4 months after the estradiol implants. Pituitary tumor tissues were collected, and cells were isolated and maintained in cultures. AF pituitary tumor cells developed stem-like cells properties and showed increased expression of cancer stem cell markers (OCT4, SOX-2, KLF-4) and EMT regulatory gene (Snail-1), and prolactin as compared to those in control-fed (AD and PF) pituitary cells. AF cells also had higher cell proliferation rate, increased invasiveness and colony formation than those in AD and PF cells, suggesting that AF pituitary cells were more aggressive cancer cells than control cells. We also found that AF cells had higher expression of Developmental Pluripotency Associated 4 (Dppa4), a gene we recently identified is overexpressed in aggressive tumor cells and in fetal alcohol exposed animals. These data are comparable to those we previously found in pituitary cells of estrogen-treated female AF rats. These data also support the view that fetal alcohol exposure programs the pituitary epithelia to a mesenchymal stem cells phenotype which is involved in induction of aggressive pituitary prolactinomas in male rats. (Supported by a NIH Grant R01 AA11591 and a Hatch Fund NJ06190)
Diabetes and Obesity (DO)
DO1. Increased Serum Lipopolysaccharide After a Short-Term High Fat Diet in Older Individuals with Obesity Compared to those with Normal Body Weight: A Randomized Crossover Study
A. Ogilvie, Y. Schlussel, PhD, J. Onishi, PhD, S. Shapses, PhD
Rutgers University
Objective: Lipopolysaccharides (LPS), also known as endotoxins, are present in gram-negative bacteria and increased serum concentrations are associated with inflammation and a high fat diet (HFD). Additionally, individuals with obesity may have greater intestinal concentrations of gram-negative bacteria that contribute to low-grade systemic inflammation. The objective of this study was to determine whether short-term higher and lower fat intakes affect serum LPS concentration in lean and obese older individuals.
Methods: In this crossover study, obese and lean older adults (50-79 years old) were randomly assigned to first 5 days of HFD (40%) or 5 days of low fat diet (LFD; 20%) followed by the other diet. These diets were separated by a 2-week washout period. Intake during each diet was recorded by participants using food records. A 5-hour mixed meal tolerance test (performed after the LFD) included a meal with 530 calories (27g; 46% fat). Recombinant Factor C assay was used to measure serum LPS (Lonza) and lipopolysaccharide binding protein (LBP) was measured by ELISA (Invitrogen Corp.). The effect of diet on LPS and LBP was assessed using paired T-tests. Two-way repeated measures ANOVA was used to assess responses to the mixed meal in lean and obese groups (SPSS v28).
Results: Participants were 64±6 years and 66% were female. Body mass index was 24.3±1.7 kg/m2 and 36.6±6.0 kg/m2 in lean (n=16) and obese (n=16) individuals, respectively. Reported fat intake during the LFD was 26±6% (7±2% SFA) and 46±6% (16±3% SFA) during the HFD (p< 0.001) and did not differ between groups. Fasting serum LPS was higher after the HFD than LFD in the obese group [HF: 0.64±0.46 EU/mL vs LF: 0.36±0.20 EU/mL, p< 0.05], but not the lean group [HF: 0.42±0.35 EU/mL vs LF: 0.34±0.22 EU/mL]. The rise in postprandial serum LPS was higher in obese than lean individuals (p< 0.05). Fasting LBP concentration did not differ between HFD and LFD, and postprandial LBP concentrations were not significantly different in obese and lean groups.
Conclusion: Individuals with obesity are more susceptible to an increase in LPS due to short-term high fat intake. Furthermore, postprandial LPS showed a marked increase only in individuals with obesity that remained elevated after 5 hours. These findings may have implications for dietary fat induced inflammation in obese populations.
DO2. Physical Exercise Improves Endocrine, Paracrine, and Autocrine Functions of Perivascular Adipose Tissue.
M. Kushkestani, M. Moghadassi
Ph.D. student of Kinesiology and Applied Physiology Department, Rutgers University, USA, Master of applied exercise physiology, Islamic Azad University
Background: Perivascular adipose tissue (PVAT) seems to play crucial adverse roles under
pathophysiological conditions such as obesity, inflammation, hypertension, and vascular diseases become dysfunctional and secrete high levels of pro-inflammatory adipokines directly on the blood vessel walls leading to endothelial dysfunction and vascular inflammation. By contrast, Exercise induces adaptations in the vessel wall that are antiatherogenic including effects on endothelial and smooth muscle cell function, along with structural remodeling of the arteries.
Objective: The purpose of this review study was to investigate the mechanism involved behind the physiological and pathophysiological effects of PVAT on vascular function as well as the effects of exercise as a modifiable factor for PVAT functions.
Methods: Three electronic databases PubMed/MEDLINE, Web of Science, and
Scopus comprehensively. Keywords that were used in this study included Prevascular Adipose tissue AND exercise AND aerobic AND resistance AND physical training. We searched for any articles that were published after 2000 and in English. Finally, 27 articles were used for this review regarding the inclusion and exclusion criteria.
Results: The results of various studies showed that physical exercise through mechanisms; 1) decreasing oxidative stress and increasing anti-oxidative defense; 2) increasing expression and level of vasodilator markers such as adiponectin and NO; 3) increasing β3-adrenoreceptor stimulation and BKCa activation; 4) decreasing expression and secretion of AGEs; 5) decreasing expression and level of vasoconstriction indices such as TNF‐α; could modify PVAT functions and prevent vascular damage.
Conclusion: Taken together, regarding the effects of physical exercise on PVAT functions in various pathophysiologic conditions (obesity, T2DM, etc.) it could be stated that exercise is a safe and cost-effective intervention to improve PVAT function and consequently prevent vascular diseases.
DO3. AI-Driven Precision Medicine: Predicting Diabetic Complications with Machine Learning Models
Fauzan Amjad
Robert Wood Johnson Medical School’s Women Health Institute
No abstract provided.
DO4. Glucagon Changes Substrates Preference in Gluconeogenesis
H. Xu, H. Kwon, Y. Wang
Rutgers University - Robert Wood Johnson Medical School
Fasting hyperglycemia in diabetes mellitus is caused by unregulated glucagon secretion that activates gluconeogenesis (GNG) and increases the use of pyruvate, lactate, amino acids, and glycerol. Studies of GNG in hepatocytes, however, tend to test a limited number of substrates at non-physiologic concentrations. Therefore, we treated cultured primary hepatocytes with three identical substrate mixtures of pyruvate/lactate, glutamine, and glycerol at serum fasting concentrations, where a different U-13C labeled substrate was substituted in each mix. In the absence of glucagon stimulation, 80% of glucose produced in primary hepatocytes incorporated either one or two 13C-labeled glycerol molecules in a 1:1 ratio, reflecting the high overall activity of this pathway. In contrast, glucose produced from 13C-labeled pyruvate/lactate or glutamine rarely incorporated two labeled molecules. While glucagon increased glycerol and pyruvate/lactate contribution to glucose carbon by 1.6- and 1.8-fold, respectively, glutamine contribution to glucose carbon was increased 6.4-fold in primary hepatocytes. To account for substrate 13C carbon loss during metabolism, we also performed a metabolic flux analysis, which confirmed that the majority of glucose carbon produced by primary hepatocytes was from glycerol. In vivo studies using a PKA-activation mouse model confirmed that most circulating lactate carbons originated from glycerol, but very little glycerol was derived from lactate carbons, reflecting glycerol’s importance as a carbon donor to GNG.
Neuroscience (N)
N1. The role of astrocyte calcium activity in the stimulant effect of alcohol
W. Evans, A. Kalelkar, S. Baskar, A. Arigbe, A. Nayak, R. Huda
Department of Cell Biology and Neuroscience, Rutgers University - New Brunswick
The dorsal striatum is an integrative nucleus innervated by sensorimotor projections from the cortex and thalamus and functionally important for both the motivation and coordination of motor activity. Though much has been learned about how diverse subtypes of striatal neurons and interneurons modulate movement, we know comparatively less about astrocyte contributions to the function of striatal circuits. In lieu of electrical excitability, astrocytes have spatiotemporally rich and robust calcium dynamics that are modulated under multiple forms of local neuronal activity. People with a higher stimulant effect and lower sedative effect of alcohol are predisposed for alcohol abuse, an effect thought to involve dysregulated dopaminergic signaling in the striatum. In animal models, ethanol also elicits a stimulatory effect in a subset of subjects, which manifests as increased locomotor activity. How astrocytes contribute to ethanol stimulation is not known. Specifically, it is unknown how striatal astrocytes respond during baseline locomotion and whether their activity contributes to the stimulant effects of ethanol.
We measured and manipulated astrocyte calcium activity to test locomotion responses of astrocytes. We surgically implanted a fiber optic in the dorsolateral striatum (DLS) of animals injected with an astrocyte specific calcium indicator. We began by characterizing the astrocytic response to locomotion in head-fixed conditions (both voluntary and forced locomotion variants) and in freely moving animals in the open field. Our results show that in head-fixed behavior animals exhibit robust striatal astrocyte activation when both voluntarily locomoting and when forced to locomote. When animals are allowed to freely locomote in the open field, we have found that calcium levels constantly fluctuate and may be associated with non-locomotor behavior as well.
We also studied ethanol’s effect on striatal astrocytes using injection of ethanol and utilizing the hPMCA calcium extruder to abolish astrocyte calcium signals. Our preliminary results show hPMCA expression was associated with increased locomotor stimulation in ethanol exposure, suggesting that astrocyte activity may play a protective role against excessive ethanol simulation. These results represent a potentially new mechanism explaining the stimulant properties of ethanol, highlighting astrocytes as a potential therapeutic target for alcohol misuse.
N2. MICROGLIAL EXOSOME INCREASES STRESS-REGULATORY PROOPIOMELANOCORTIN NEURONAL DEATH IN THE DEVELOPING HYPOTHALAMUS VIA ACTIVATING MONOCYTE CHEMOATTRACTANT PROTEIN 1
P. Tarale, S. Mukherjee, S. Chaudhary
Rutgers Endocrine Research Program, Department of Animal Sciences, Rutgers University, New Brunswick, NJ 08901, USA
Microglia, a type of CNS immune cells, are shown to participate in ethanol induced neuronal death in the fetal brain to induce fetal alcohol syndrome. We have previously shown that microglial cells are involved in killing of proopiomelanocortin (POMC) neurons that leads to the development of abnormal stress response in fetal alcohol exposed offspring. How microglia kill these neurons is not wells understood. We have recently shown that microglia participate in secreting extracellular vesicles (exosomes), composed with genetic material, DNA, mRNA, and proteins, to communicate with proopiomelanocortin neurons. In this study, we determined if exosomes secreted from ethanol stimulated microglia triggers POMC neuronal cell death by activating the chemokine system in them. Exosomes were isolated from hypothalamic tissues collected from postnatal rats (both males and females) fed daily (PND2-PND6) with 2.5 mg/kg ethanol or control milk formula or from postnatal rat’s hypothalamic microglia grown in culture and activated with 50 mM ethanol or vehicle for 24 h. Magpix and ELISA results shows significantly elevated level of chemokines; MCP1/CCL2 and MIP1-alpha/CCL3 within both in-vitro and in-vivo microglia derived exosomes activated by ethanol. Nucleosome assay, immunocytochemistry and qPCR results show that treatment with RS504393; a blocker of MCP1/CCR2 signaling, attenuated the primary POMC/beta-endorphin neuronal apoptosis. In-vivo subcutaneous injection of RS504393 (1mg/kg) in postnatal rats fed with 2.5 mg/kg ethanol prevented POMC/beta-endorphin neuronal death. These data suggest that the chemokine system involving MCP1/CCL2 participates in exosome.
N3. Effects of Mindfulness-Oriented Recovery Enhancement in Women with Opioid Use Disorder
S. Ray, A. Vadrevu, J. Bhanji, P. Dooley Budsock, M. Delgado, E. Garland
Rutgers School of Health Professions
Objective: Mindfulness-Oriented Recovery Enhancement (MORE) is an efficacious program to aid recovery from substance use disorder. This study in a pilot sample of individuals in treatment for opioid use disorder (OUD) characterizes the immediate effects of a brief MORE guided meditation session as well as longer-term changes after the MORE program.
Design: Twelve female participants in residential treatment for OUD completed an 8-week MORE program. Participants completed assessments before and after the MORE program, which included a 10-minute guided MORE meditation during functional magnetic resonance imaging (fMRI). Changes in mood and opioid craving were assessed relative to each guided mediation. Functional connectivity (i.e. correlated fMRI signal) between regions in a hypothesized affect regulation network was measured during the meditation to assess change in brain network function after MORE.
Results: Nine participants completed all measurements. Mood improved significantly after each 10-minute guided mediation. Mood also increased from before to after the eight-week MORE program, though the change was not statistically significant. The 8-week MORE program decreased opioid craving in individuals who were still reporting opioid craving at baseline. Connectivity between left ventromedial prefrontal cortex and left amygdala was significantly greater after the MORE program. Connectivity between left ventrolateral prefrontal cortex and left nucleus accumbens showed a similar significant increase after the MORE program.
Conclusions: This pilot study characterizes potential benefits of MORE for prefrontal-subcortical neural connectivity and mood improvement related to a single meditation session in women with OUD. This characterization can guide investigations in larger samples.
N4. Network Meta-Analysis on the Effects of Ethanol Exposure on the Onset and Progression of Alzheimer’s Disease through Investigation of Prenatal, Adolescent, and Adult Genome Samples
R. J. Kim , W. Huang, M. Bishir, S. L. Chang, Ph.D.
Institute of NeuroImmune Pharmacology, Department of Biological Sciences, Seton Hall University, South Orange, NJ
Alcohol consumption can result in the inflammation of various organs such as the intestines, liver, and brain. With regards to the brain, Alzheimer’s Disease (AD) is a degenerative disorder which can result from inflammation and accumulation of amyloid proteins and neurofibrillary tangles. As of 2021, approximately 6.2 million Americans aged 65 and older live with AD, and this number is anticipated to double within the following decades. Yet, among many genetic and environmental determinants, alcoholism may play a role in neurological damages and disease. To understand the effects of alcoholism in the changes of neuronal gene expression over time, and how such changes may promote the onset of AD, this study has conducted a network meta-analysis on the onset and progression of AD through investigation of prenatal, adolescent, and adult brain tissue samples. In studying prenatal cortical samples with gestational age of 15 to 18 weeks, CLC 22 was used to analyze alcohol-exposed cortical genomes against controls. Differential analysis identified a total of 1,049 differentially expressed genes (DEG). These 1,049 DEGs were then studied on IPA through Core Analysis to find 283 most associated canonical pathways (p-value<0.05), in which the top pathway was Hepatic Fibrosis (p-value 1.07E-27). CLC 22 was also used to compare serum microRNA of alcoholic adolescents aged 18 to 21, against non-alcoholic adolescents of the same age cohort. 146 DEGs were identified, and IPA’s Core Analysis on these molecules revealed 39 most associated pathways (p-value<0.05). Lastly, CLC 22 was used to compare 60+ aged post-mortem white matter transcriptomes of alcoholic and non-alcoholic individuals. 2851 DEGs were identified, and IPA’s Core Analysis revealed 191 most associated canonical pathways (p-value 0.05). Upon studying the DEGs within a biological connectivity network on IPA and simulating exposure of these molecules to ethanol (EtOH), it was found that EtOH exposure results in a synergistic effect between the DEGs to cause activation of Amyloid Precursor Protein (APP) expression, and onset of AD (p-value 4.47E-2).
N5. MOLECULAR MECHANISMS UNDERLYING COVID-19 MODULATION OF DRUG ADDICTION RELATED SIGNALING PATHWAYS IN THE BRAIN
M. Bishir, W. Huang, S. L. Chang
Institute of NeuroImmune Pharmacology, Department of Biological Sciences, Seton Hall University, South Orange, New Jersey
Reports have revealed an increase in substance use and drug overdose cases in the US since the COVID-19 pandemic was declared as a national emergency in March 2020. There has been a 23% increase in alcohol abuse and a 16% increase in drug abuse for people who had consumed those substances before the pandemic. In addition to increased consumption among the long-time substance users, isolation and COVID-19 related fears have been associated with limited access to detoxification centers and psychological distress, playing a major role in the increase of drug and alcohol use. We hypothesize that SARS-CoV-2 virus modulates drug addiction related signaling pathways in the brain. We identified publicly available datasets from GEO database containing postmortem brain samples from COVID-19 patients. Differentially Expressed Genes (DEG’s) from GSE236562 (PFC-BA9), GSE188847 (PFC); GSE159812 PFC (single cell RNA-seq-PFC) and amygdala of COVID-19 brains were identified through GEO2R and CLC-12. DEG’s were imported to Ingenuity Pathway Analysis-(IPA) to perform core analysis and to identify the signaling pathways and upstream regulators associated with DEG’s from the PFC and amygdala of COVID-19 patients. Our “Comparison Analysis” of the signaling pathways found the activation of NeuroInflammation signaling pathway (NISP), and inhibition of Dopamine-DARPP32 Feedback in cAMP Signaling, Opioid signaling pathway and inhibition of Glutamate signaling and Synaptic plasticity in the PFC and amygdala of COVID-19 patients. We also identified 31 common upstream regulators, including IFNG, IL1B, IRF1, JUN, STAT1, STAT3, and TNF, which were also found to be involved in NISP. Overlying the DEG’s onto the upstream regulator-NISP network map further confirmed the activation of NISP. Taken together, our results showed that COVID-19 modulates pathways related to drug addiction, including activation of NISP, and inhibition of dopamine and opioid signaling and synaptic plasticity in the PFC and amygdala of COVID-19 patients.
N6. GLP-1 signaling in the hypothalamic-brain stem descending circuit regulates energy homeostasis.
L. Wang
Department of Neuroscience and Cell Biology, Child Health Institute of New Jersey, Rutgers University Robert Wood Johnson Medical School
Central nervous system (CNS) control of metabolism plays a pivotal role in maintaining energy homeostasis. Glucagon-like peptide 1 (GLP-1, encoded by Gcg), secreted by a distinct population of neurons located within the Nucleus Tractus solitarius, suppresses feeding. Central and peripheral GLP-1 work independently to suppress feeding. However, the cellular and circuit mechanisms mediating endogenous GLP-1 action in the CNS are still poorly understood, mainly due to diverse neuronal subtypes and complex central neuronal connectivity. We previously found that NTS GLP-1 projection to the paraventricular hypothalamic nucleus (PVN) enhances glutamatergic synaptic transmission, which is sufficient to suppress food intake, and ablation of PVN GLP-1R causes overeating and obesity. Here we investigate the impact of PVN GLP-1R neuronal descending neuronal ensemble, particularly the brain stem dorsal vagal complex (DVC), in energy homeostasis. Using multiple anterograde pathways tracing, we found PVN GLP-1R neurons form synapses with DVC neurons release glutamate and are regulated by GLP-1 presynaptically. Using retrograde Patch-Seq, we revealed the complex neuronal identity of PVN GLP-1R neurons, including oxytocinergic and non-oxytocinergic vGlut2/GLP-1R neurons. Chemogenetic activation of the PVN-to-DVC GLP-1R circuit is sufficient to suppress food intake in different motivational states Loss-of-function studies by blocking the PVN GLP-1R-to-DVC synaptic release or ablation of GLP-1R in the PVN-to-DVC projecting neurons increases food intake and cause type 2 diabetes, including obesity, elevated blood glucose, and deficits in glucose metabolism. These findings suggest that GLP-1 signaling regulates the PVN-to-brain stem descending circuit and regulates energy homeostasis.
N7. Top-down modulation of autonomic arousal by the anterior cingulate cortex
N. Chintalacheruvu, A. Kalelkar
Rutgers-NB, CBN department, Huda Laboratory
Hyperactivity of arousal-mediating circuits are implicated in a variety of neuropsychiatric disorders. Therefore, insight into the arousal system of the brain could lead to a better understanding of the neurological basis of these disorders. Subcortical structures such as the locus coeruleus, hypothalamus, and periaqueductal gray (PAG) are known to be involved in the regulation of autonomic arousal but much less is known about the role of higher level cortical circuits. We investigated the role of a major subdivision of the prefrontal cortex, the anterior cingulate cortex (ACC), in modulating autonomic arousal. Pupil size, heart rate, and movement were used as non-invasive measures of arousal. To test how ACC activity contributes to arousal levels, we simultaneously measured arousal and recorded fiber photometry signals from mice virally expressing GCaMP8m. Aligning these signals to the onsets of pupil dilations revealed that pupil dilations were preceded by increases in ACC calcium activity. Further, ACC activity was associated with heart rate increases. To better understand how the ACC can directly modulate arousal, we optogenetically manipulated ACC pyramidal neuron activity. We found that photostimulation of pyramidal neurons in mice expressing CaMKII-ChR2 caused pupil dilations that were time-locked to activation. ACC activation also led to increased heart rate and movement. ACC inactivation via photostimulation of GABAergic neurons caused pupil constrictions. Additionally, closed-loop optogenetic inactivation of the ACC using DeepLabCut Live suppressed ongoing pupil dilations. Vasoactive intestinal peptide-expressing (VIP) interneurons exert disinhibitory control over pyramidal neuron activity by inhibiting other inhibitory cells. We tested whether and how VIP neurons are involved in ACC modulation of arousal. Optogenetic VIP neuron stimulation caused pupil dilations. Next, we investigated a possible ACC to periaqueductal gray (PAG) projection pathway that may be involved in arousal modulation. We used fiber photometry to record calcium activity from ACC axons in the PAG. Aligning calcium activity to onsets of pupil dilations revealed pupil dilations were preceded by increases in ACC-PAG axon activity, similar to what we found in ACC neurons. We also found that optogenetic stimulation of ACC axons in the PAG caused pupil dilations. Together these findings indicate that the ACC modulates autonomic arousal, possibly via projections to the PAG. We are currently evaluating how ACC VIP neurons regulate outflow of information to the PAG for top-down modulation of arousal.
N8. Deciphering GLP-1 Release Dynamics and Feeding Behavior with a Novel Fluorescent Sensor
F. Luo, Y. Lu, V. Mirabella, R. Savani, L. Wang, Z. Pang
Child Health Institute of New Jersey
Glucagon-like peptide 1 (GLP-1), is secreted by intestinal L-cells and a subset of hindbrain NTS neurons. it plays a crucial role in feeding and energy homeostasis. However, the release patterns of endogenous GLP-1 in the brain and their correlation with animal behavior remain poorly understood. The lack of molecular tools capable of directly detecting real-time GLP-1 poses a significant challenge. In this study, we present a novel and highly sensitive optical sensor called Reporter for Transmission mediated by G-protein coupled Receptor (RTGR), specifically designed for GLP-1 detection.
Using fiber photometry combined with optogenetic stimulation of GLP-1-expressing NTS neurons, we demonstrated the high sensitivity of the GLP-1 RTGR sensor in detecting GLP-1 release in mice. Moreover, through overnight photometry recordings in freely moving mice, we observed a correlation between endogenous GLP-1 release and naturalistic feeding behavior in the paraventricular nucleus of the hypothalamus (PVN). Following each feeding bout, GLP-1 levels in the PVN showed a significant increase.
These findings shed light on the physiological role of central endogenous GLP-1 release in feeding and other naturalistic behaviors. Ongoing investigations aim to further validate these results and explore the implications of GLP-1 release in food intake behavior. The development of the RTGR sensor provides a valuable tool for studying GLP-1 dynamics in real-time, facilitating deeper insights into the mechanisms underlying GLP-1's effects on feeding and energy regulation.
N9. MIR-383 AND MIR-384 SUPPRESS PROOPIOMELANOCORTIN GENE EXPRESSION IN THE HYPOTHALMUS: EFFECTS OF EARLY LIFE ETHANOL EXPOSURE
O. Gangisetty, S. Chaudhary, P. Tarale, M. Cabrera, D. K. Sarkar
Endocrine Research Program, Dept of Animal Sciences, Rutgers University, New Brunswick, NJ-08901, USA
Prenatal alcohol exposure (PAE) causes several abnormalities in neuroendocrine functions including stress hyperresponses. Studies suggest a loss of the proopiomelanocortin (POMC) neuronal control of hypothalamic pituitary adrenal axis (HPA) function is critical for the PAE induced stress hyperresponse. We have previously shown PAE alters Pomc gene expression via epigenetic mechanism involving increased promoter DNA methylation. MicroRNAs are another type of epigenetic modifiers that regulate gene expression in a sequence specific manner. In this study we tested whether microRNAs (miRs) contribute to the PAE-induced suppression of Pomc gene. We fed POMC-EGFP mice with a milk formula containing ethanol (11.34%) yielding a total ethanol dose of 2.5g/kg (AF) or pair-fed equal caloric milk formula (PF) or left in the litter (AD) during early postnatal day (PND) 2-6. Genome wide microRNA expression profile identified 40 miRNAs significantly altered in hypothalamic tissues of AF mice. In silico analysis further identified miRNA-383, -384, and -488 have putative binding sites at the POMC 3’UTR. Data show that AF mice had reduced Pomc expression in the hypothalamus compared to AD and PF control mice. While POMC targeted miR-383 and miR-384, but not miR-488, levels were increased in the hypothalamus of AF mice as compared to AD and PF mice. We further established the role of these miRs in regulation of Pomc using miR over expression and knock down approaches in mHypoA-POMC/GFP neurons in cell cultures. These cells when transfected with miR-383 or 384 mimic oligos robustly increased the expression of corresponding miRs and significantly reduced Pomc expression compared to negative control oligos. Administration of miR-383 or 384 inhibitor oligos suppressed ethanol stimulated miR-383 or 384 expression and restored POMC expression in AF mice. Additionally, antimir-383 or 384 oligos effectively reduced the corresponding miRs compared to negative control oligos. Also, Pomc expression was significantly increased with antimir-383 and 384 oligos. We further evaluated the specificity of these miRs in repressing Pomc expression by specifically targeting its 3’UTR. We cloned mPomc 3’UTR downstream of luciferase in a reporter vector. We found that miR-383, miR-384 mimic oligo effectively reduced the luciferase activity of the reporter vector containing Pomc 3’UTR suggesting that miR-383 or miR-384 represses Pomc expression by targeting its 3’UTR. These data suggest that PAE reduces Pomc expression and alters stress axis function possibly by enhancing the expression of miR-383 and miR-384 in the hypothalamus. (Supported by NIH grant R01AA025359).
N10. Epigenetic Inheritance and Disease Susceptibility of fetal Alcohol Exposed Offspring in the Male Germline: Insights from Transgenerational Analysis of Fischer Rats
L. Gerlikhman, A. Palagani, O. Gangisetty, D. Sarkar
Endocrinology Program, Rutgers, The State University of New Jersey, New Brunswick, NJ, Molecular Neuroscience of Alcohol and Drug Abuse Research Training, Rutgers, The State University of New Jersey. Department of Animal Sciences, Rutgers, The State University of New Jersey
Background: Epigenetic modifications play a critical role in gene regulation and inheritance. Understanding how these modifications are transmitted across generations and their potential impact on disease susceptibility is a fundamental question in biology. In this study, we conducted a comprehensive analysis of epigenetic variations in genomic DNA isolated from frozen sperm pellets of alcohol or pair fed (control) Fischer Rats, with a specific focus on the male germline (AFM), spanning multiple generations (F1, F2, and F3).
Methods: Genomic DNA isolation, library preparation, high-throughput sequencing, and advanced bioinformatic and statistical analyses were conducted. Genomic DNA was extracted from frozen sperm pellets, and libraries were prepared for methylation sequencing using the Illumina NextSeq 550 instrument. Rigorous quality control, read processing, alignment to the rat genome (Rnor_6.0), and methylation call extraction were performed. Differential methylation analysis identified statistically significant variations in methylation patterns.
Results: Our analysis revealed intriguing patterns of epigenetic variations transmitted across generations within the male germline of AFM. Specifically, 1117 hypermethylated and 1456 hypomethylated genes were identified and consistently transmitted from F1 to F3. To identify potential diseases passed down through the male germline, genomic locations, such as promoters, introns, exons, and intergenic regions were characterized for genes hypermethylated or hypomethylated on the X or Y chromosome of AFM. Notably, 11 genes were hypermethylated on the X chromosome of AFM and one on Y, while 4 genes were hypomethylated on the X chromosome. Disease Ontology Analysis: Genes hypermethylated or hypomethylated on the sex chromosomes (XY) of the male germline were associated with intellectual disability, autism, neurodegenerative diseases, and other disorders. These findings suggest a potential role of epigenetics in the inheritance of disease-related traits in the male germline.
Conclusion: The identification of hypermethylated and hypomethylated genes consistently transmitted across generations in the sex chromosomes underscores the importance of epigenetic regulation in inheritance. Moreover, associations with disease-related terms in our disease ontology analysis, including those specific to the male germline, highlight the potential role of transgenerational epigenetics in disease susceptibility.
N11. Involvement of Sigma-1 Receptor in Ethanol Modulation of Pain Sensitivity
J. Zhang, M. Bishir, S. L. Chang
Institute of NeuroImmune Pharmacology and Department of Biological Sciences, Seton Hall University, South Orange, NJ, USA
Sigma 1 receptor (S1R) is a membrane receptor that mediates the action of various additive substances including cocaine, morphine, and methamphetamine. Ethanol (EtOH) is involved in modulating various types of pain. Studies have shown short time use of alcohol use produces analgesic effects while persistent alcohol use develops chronic pain. We hypothesize S1R is involved in EtOH modulation of analgesia. We use QIAGEN Ingenuity Pathway Analysis (IPA) to construct a connectivity map to test our hypothesis. EtOH was found to activate S1R via 28 intermediary molecules. We conducted a downstream effect analysis of these intermediary molecules to quantify the effect of EtOH on S1R. Based on these data, a pathway connectivity map from EtOH to various nodes related to pain including discomfort, acute pain, chronic pain, hyperalgesia, hypoalgesia, neuropathic pain, and inflammatory pain was generated. Differentially expressed genes (DEGs) of various brain regions of binge-ethanol-treated mice compared to naïve mice were collected from the GSE93311 dataset. DEGs in the prefrontal cortex of post-mortem humans with either alcohol abuse (short time use of alcohol) or alcohol dependence (persistent use of alcohol) compared to matched controls were collected from the GSE49376 dataset. These DEGs were overlayed onto the constructed pathway established using QIAGEN IPA. These analyses revealed that S1R expression was regulated in different types of pain. With short time binge alcohol, increased S1R expression appeared to inhibit acute pain signaling in the prefrontal cortex; while decreased S1R expression seemed to activate hyperalgesia signaling in the nucleus acumens shell. SIR expression was lower in those with alcohol dependence than those who abused alcohol suggesting that SIR increased analgesia in acute alcohol abuse. The neuropathic pain signaling pathway in dorsal horn neurons, NGF signaling, and opioid signaling were among the enriched pathways for alcohol dependence. Acute EtOH activated S1R leading to analgesic effects.
N12. Biological Self-Communication in Relation to Early Life Stress
M. Whedon, M. Lewis
Rutgers Robert Wood Johnson Medical School
Effective communication between biological systems in the context of stress is important for adaptive coping and well-being. During stress, the hypothalamic pituitary adrenal (HPA)-axis system is activated, resulting in the release of glucocorticoids by the adrenal glands. By opposing the effects of cortisol, the co-release of dehydroepiandrosterone (DHEA) alongside of cortisol is thought to facilitate the HPA system return to homeostasis (Romeo, 2005). In contrast to adults, research has suggested these hormones are tightly coupled in response to stress during adolescence. Other work suggests that cortisol-DHEA coupling may be adversely impacted by early life stress. The purpose of this study is to examine the relations between cortisol and DHEA reactivity to stress in a sample of adolescents characterized by elevated environmental risk. In this study, youth were participants in a longitudinal investigation of the effects of prenatal cocaine exposure (PCE) on development. At 11.5, 13.5 and 17 years of age youth participated in standardized paradigms to elicit a stress response (social evaluative threat); saliva samples were provided before and after the stressor; DHEA and cortisol concentrations were later determined using commercially available high-sensitivity EIA kits. At each timepoint, the significance of within-person differences in both hormones were assessed separately for exposed and unexposed males and females. Latent change score models were used to examine the associations between pre- and post-stressor hormone levels, controlling for effects of time and body composition. On average, pre-stressor DHEA and cortisol levels are higher than post-stressor levels across timepoints. However, by 13.5 years, these changes are not significant among PCE males. Additionally, across timepoints, only unexposed youth show significant covariation between pre-stressor cortisol and DHEA. Results are ongoing, but suggest that PCE may disrupt the capacity for DHEA to modulate the cortisol response to stress. Future studies should consider collect multiple samples so that within-person associations between these hormones can be estimated.
N13. Stress Reactivity and Substance Use across the Transition to Adolescence
M. Whedon, M. Lewis, D. Dick
Rutgers Robert Wood Johnson Medical School
Although adolescence is when most people first try alcohol and other psychoactive substances, early initiation of substance use is implicated as a risk factor for addiction and may adversely impact the developing nervous system. The stress response system is thought to become recalibrated during adolescence and animal work suggests that alcohol use during the adolescent period may alter the neural systems that regulate the stress response (e.g., Syzmanska et al., 2011; Allen et al. 2011). Although many studies with humans have suggested that adolescent substance use is associated with cortisol hyporesponsivity to stress, the directionality of effects is unclear. The purpose of this study was to investigate whether early initiation of substance use in adolescence is a cause or consequence of aberrant stress-reactivity. Participants were part of a longitudinal study examining the effects of prenatal cocaine exposure (PCE) on development. Between 13 and 15 years, youth reported on their substance use with the Youth Risk Behavior Survey (N=156, 54% male, 39% exposed). Stress reactivity was assessed at 11.5 and 17 years using age-appropriate paradigms (peer competition, Trier Social Stress Test) and salivary cortisol assay. Latent change score models were used to assess the bidirectional associations between stress reactivity and substance use, with biological sex and PCE modeled as predictors. Results indicated that pre-stressor cortisol is stable from 11.5 to 17 but cortisol reactivity is not, and substance use is not associated with cortisol at either timepoint (see Figure 1). PCE and sex do not account for significant variance in preadolescent cortisol, but prenatally exposed males are more likely to exhibit increases in cortisol to stress at 17 compared to their non-exposed peers (see Figure 2). PCE may interact with hormonal changes associated with puberty to influence stress reactivity in adolescence, but further work is needed to better understand sex differences and the role of substance use.
N14. Postpartum behavioral deficits caused by disruption in microtubule stability
S. Gonzalez-Salinas, I. Fuentes, Y. Morishita, D. Barker, S. Uchida, G. Shumyatsky
Department of Genetics, Rutgers, Busch Campus, Department of Cognitive Function and Pathology, Institute of Brain Science, Nagoya, Japan, Department of Psychology, Rutgers, Busch Campus, SK project, Medical Innovation Center, Kyoto University Graduate School of Medicine, Kyoto, Japan.
Postpartum depression (PPD) affects mother’s well-being, impacts her ability to provide proper childcare, and detrimentally affects the emotional and cognitive development of the progeny. Studies in mothers with PPD and in mouse models of gestational stress show that the behavioral changes postpartum are related to disrupted activity in corticolimbic regions, such as the medial prefrontal cortex (mPFC), but the mechanisms of these processes remain unclear. Therefore, we have examined whether changes in microtubule stability in the mPFC regulate maternal behaviors. We found that mothers with deficient microtubule stability have decreased pup survival, deficient maternal care (longer pup retrieval to the nest) and exhibited more anxiety- and depressive-like behaviors. We showed that gestational corticosterone induced alterations in microtubule stability in the PFC. To examine neural connections between corticolimbic and key maternal brain areas, we traced PFC projections to the medial preoptic area (mPOA) and quantified neuronal activity following pup retrieval. We show that more PFC cells projecting to the mPOA become active following pup retrieval in mothers compared to virgin females. Our data show that microtubule stability in corticolimbic circuits plays a key role in postpartum behaviors and that alterations in the function of the mPFC could explain deficient maternal care.
Nutrition, Metabolism, and Mental Health (NMMH)
NMMH1. Sex differences in cognition from perinatal exposure to organophosphate flame-retardants in male and female adult offspring
K. Wiersielis, R. Mukadam, J. Early, N. Knox, T. Degroat, T. Roepke
Rutgers University
Endocrine disrupting compounds (EDCs) are compounds found in our environment that interrupt typical endocrine function. A particular group of EDCs are flame-retardants due to their interaction with steroid and nuclear receptors. Humans are consistently exposed to flame-retardants daily as they are used in everyday items such as plastics, clothing, toys, and electronics. A commonly used class of flame retardants are the organophosphate flame-retardants (OPFRs), which have been shown to alter adult behavior in rodent species after developmental exposures. We have previously identified that males perinatally exposed to OPFRs had a trending reduction in locomotion and females had a significant increase when in the open field task. In addition, we detected OPFR males exhibited anxiolytic-like behavior in the elevated plus maze. However, the effects of perinatal OPFR exposure on cognition in the adult offspring are underexplored. Here we evaluate cognitive behavior using the Y-maze, spatial object recognition (SOR), novel object recognition (NOR), and the Barnes maze in male and female adult offspring that were perinatally exposed to a mixture of OPFRs (tris(1,3-dichloro-2-propyl)phosphate, triphenyl phosphate, tricresyl phosphate) or vehicle. In the Y-maze, OPFR-exposed males exhibited a reduction in the number of entries to the unknown arm and OPFR-exposed females exhibited an increase, suggesting an impairment and enhancement in memory, respectively. In the SOR, a similar pattern emerged such that OPFR males exhibited a decrease in time spent with the displaced object and OPFR females showed an increase. In both the NOR and the Barnes maze, we did not observe any treatment or sex differences. Our results demonstrate sex- and exposure-dependent effects of perinatal OPFR exposure on cognition in male and female adult mice, suggesting developmental neurotoxicity. Future studies will examine the influence of perinatal OPFR exposure on the long-term potentiation of pyramidal cells in hippocampal CA1 and acetylcholine excitatory synaptic transmission.
NMMH2. The interactions of maternal diet and exposure to flame retardants on offspring metabolism and behaviors
C. Rojas, A. Yasrebi, S. Adams, S. Anthony, L. Aleksunes, T. A. Roepke
Joint Graduate Program in Toxicology, Department of Animal Sciences, SEBS
We previously demonstrated that maternal exposure to organophosphate flame retardants (OPFRs) alters adult offspring metabolism and behavior. Other studies have shown that maternal obesity can also alter offspring behavior by disrupting the formation of the blood brain barrier during development. The goal of this project is to understand the potential synergistic effects of maternal obesity and OPFR exposure on fetal development which can lead to greater metabolic and behavioral disruption in adulthood. We fed dams either a low-fat (LFD) or high-fat diet (HFD) for 8 weeks, conducted metabolic phenotyping including body weight, fat mass, glucose tolerance, and mated dams with untreated males. Dams were dosed with a dried peanut butter and OPFR (TPP, TCP, TDCPP, 1 mg each/kg) oil mixture once daily from GD7 to PND14. Pups were allowed to mature until 8 weeks before behavioral and metabolic phenotyping began. Anxiety-like behaviors were evaluated using open field test, elevated plus maze, and light dark box. All three tests indicated that LFD OPFR treated mice exhibited an anxiogenic phenotype. In addition, female HFD OIL treated mice spent more time in the unknown arm of the Y-Maze test than female HFD OP treated mice and male HFD OIL, further suggesting that both maternal diet and OPFRs affect hippocampal dependent memory. Preliminary CLAMS data indicates that maternally dosed OP female offspring are expending less energy during nocturnal hours than their oil counterparts and also exhibit less locomotion and wheel running. Future endpoints will include medial basal hypothalamic and liver gene expression of PND0 and PND14 mice and GCMS detection of OPFRs in the brains of PND0 neonates.
NMMH3. Effects of parathyroidectomy on inflammatory cytokines and vitamin D metabolites in patients with primary hyperparathyroidism
X. Wang, L. Meng, S. A. Shapses
Division of Endocrinology, Department of Medicine, Rutgers University-Robert Wood Johnson Medical School, New Brunswick,NJ. Department of Nutritional Sciences, Rutgers University, New Brunswick, NJ, Department of Nutritional Sciences, Rutgers University, New Brunswick, NJ. Division of Endocrinology, Department of Medicine, Rutgers University-Robert Wood Johnson Medical School, New Brunswick, NJ
Objective: Primary hyperparathyroidism (PHPT) is companied by decreased 25-hydroxyvitamin D (25OHD) and elevated cytokine levels. Interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) are inflammatory cytokines associated with increased cardiovascular risk factors and morbidity. And these cytokines are also found to be induced by high intact parathyroid hormone (iPTH) level. This study investigated the effect of parathyroidectomy (PTX) on inflammatory profiles and vitamin D metabolites in patients with PHPT.
Methods: Fifty patients with newly diagnosed PHPT were recruited in the study. All of them underwent PTX whereas 28 patients returned for 3-month follow-up visit after surgery. Serum samples were collected before and 3-month after PTX. Serum levels of calcium, iPTH, vitamin D binding protein (DBP), total and free 25OHD, MCP-1, IL-6, interleukin-10 (IL-10), and C-reactive protein (CRP) were measured. Thirty age and BMI matched healthy subjects were included as controls. Student and paired t-test, and correlation coefficients were used for statistical analysis.
Results: Prior to PTX, serum levels of DBP, total and free 25OHD were lower, while IL-6 and MCP-1 were higher in PHPT patients compared to that of controls (p<0.01). After PTX, serum iPTH and calcium returned to normal range (PTH: 119.2±39.8 to 43.2±24.2 pg/mL; Ca: 11.0±0.5 to 9.6±0.4 mg/dL; p<0.001). Meanwhile, serum total and free 25OHD and DBP levels were increased after PTX surgery (p<0.01). Levels of IL-6 and MCP-1 were decreased from 395.3±198.9 to 288.9±129.4 pg/mL and 15.9 ± 8.6 to 2.9 ± 1.6 pg/mL, respectively (p<0.05), while IL-10 and CRP levels remained unchanged after PTX. Circulating IL-6 and MCP-1, not CRP or IL-10, were positively correlated with iPTH (r=0.24 and r=0.4, respectively, p<0.05). The DBP was inversely correlated with PTH and IL-6 (p<0.01), but not with any other cytokines.
Conclusion: In patients with PHPT, elevated iPTH levels may upregulate the production of MCP-1 and IL-6, not CRP or IL-10, and downregulate DBP production, which were all normalized by successful PTX surgery. As expected PTX markedly decreased in serum iPTH and calcium, and there was an increase in total and free 25OHD levels. Our results suggest that PTX may have additional benefit effects on cardiovascular risk factors by decreasing IL-6 and MCP-1. The role of IL-6 and MCP-1 on DBP and vitamin D metabolites in patients with PHPT remains unclear and further studies are needed.
NMMH4. NETWORK META-ANALYSIS ON THE MECHANISMS UNDERLYING ALCOHOL-INDUCED THYMIC ATROPHY
T. Rengifo, M. Bishir, S. L. Chang
Institute of NeuroImmune Pharmacology, Seton Hall University
Thymic atrophy is characterized by the loss of thymocytes and the destruction of thymic architecture which eventually leads to a significant decrease in naïve T cells, limited T cell receptor diversity and compromised the overall immunity. Several environmental factors are known to induce thymic atrophy including alcohol misuse. Specifically, chronic alcohol use has been linked to decreased thymus size over time. Despite the known link between long-term alcohol use and thymic atrophy, the mechanism underlying alcohol induction of thymic atrophy has been understudied. The goal of the current meta-analysis study was to identify molecules and signaling pathways underlying alcohol-induced thymic atrophy using network meta-analysis in-silico in hope to develop therapeutical strategy to battle the thymic tissue damage in alcohol misuse. By taking the first approach using QIAGEN Knowledge Base (QKB), the Ingenuity Pathway Analysis (IPA) “Grow” tool and “Path Explorer” tool were used to identify the overlaying molecules between those associated with EtOH and with Atrophy of thymus gland. The second approach was involved in taking Mouse Genome Informatics (MGI) to characterize the molecules associated with the thymic atrophy. Together, 13 commonly shared molecules between EtOH and Atrophy of thymus were identified. The IPA “Molecular Activity Predictor” (MAP) tool was used to simulate EtOH mimicking exposure to alcohol leading to elevation of lipopolysaccharide level, expression of IL6, TNF, NFKBIA, and STUB1 and inhibition of expression of STAT1, EGR1, BCL2, and MAPK14 concurrently causing the increment of Atrophy of Thymus Gland. These meta-analysis data revealed key molecules underlying the causal relationship between EtOH levels and increment of thymic atrophy leading to the eventual deterioration of the overall immunity. Lastly, two GSE datasets have been collected to confirm the established pathways from EtOH to thymic atrophy. Using IPA, we were able to analyze the gene expression data and validate these possible mechanisms.
NMMH5. Retinoid metabolism during the transition from fasting to re-feeding
Y. Kim, D. Champaneri, E. Madonna, L. Quadro
Department of Food Science and Rutgers Center for Lipid Research, Institute of Food Nutrition and Health, Rutgers University, New Brunswick, NJ, USA
The essential nutrient vitamin A maintains the health of the body. Its active form, retinoic acid, functions as a transcriptional regulator of hundreds of genes. In mammalian tissues, retinoic acid is synthesized from retinol, which, upon release from the liver, circulates bound to retinol-binding protein (RBP) and transthyretin (TTR) to be distributed to the target tissues. Recent literatures suggest that serum RBP levels and hepatic retinoic acid concentrations are modulated during the transition from fasting to re-feeding. Moreover, our laboratory showed that retinoic acid regulates energy homeostasis in the heart. Here we investigated whether the levels of 1) cardiac retinoic acid and 2) the serum retinol-RBP-TTR complex fluctuate during the above-mentioned transition. We addressed these questions in WT mice fasted for 16 hours or fasted for 3 hours and re-fed for 1 or 3 hours. Our data indicate that 1) cardiac retinoic acid levels may be attenuated during refeeding, as already shown in the liver; and 2) refeeding results in increased serum levels of retinol, RBP and TTR. Interestingly, hepatic vitamin A stores were increased during fasting and normalized rapidly upon re-feeding. Overall, these findings show substantial perturbation of whole-body vitamin A homeostasis during the transition from fasting to re-feeding and warrant further studies to understand the molecular mechanisms behind these changes and their physiological implications.
NMMH6. Interventions for Stress Reduction and Developmental Improvement in Premature Infants in the Neonatal Intensive Care Unit (NICU)
N. Gonzalez, MD, G. Bachmann, MD, MMS
Women's Health Institute Rutgers Robert Wood Johnson Medical School
INTRODUCTION: Premature infants face a higher risk for complications including feeding issues, developmental delays and increased stress levels. Going through the experience of having a premature baby requiring a stay at the Neonatal Intensive Care Unit (NICU) also poses a challenge to parents, who are at increased risk for depression, anxiety and higher stress levels. This highlights the importance of utilizing non-invasive interventions, such as early skin-to-skin, music therapy or early oral motor interventions, as they may positively impact the well-being of both premature infants and their family/caregivers.
METHODS: The database used was PubMed, including all articles published in the English language within the last five years. A combination of the following keywords was utilized: “prenatal birth”, “premature babies”, “stress level”, “environmental stressors” and “Neonatal Intensive Care Unit”.
RESULTS: Five studies were identified: four randomized controlled trials and one followed a baseline-response design. One study investigated the effects of skin-to-skin contact after delivery compared to visual contact only. The study reported positive outcomes for mothers, including decreased postpartum depression and improved bonding with their infants, while also having a positive impact on premature infants through stress reduction. Another study assessed the benefits of skin-to-skin contact on stress level in preterm infants in the NICU. Results indicated an increase in oxytocin levels and a decrease in cortisol levels, suggesting that this intervention can reduce stress. Two studies evaluated the impact of creative music therapy on stress and anxiety levels in parents of premature infants. Parents who received music therapy reported reduced anxiety, depression and stress, along with improved bonding with their infants. One study investigated the effect of early infant oral stimulation in premature babies. The findings indicated that infants who received early oral stimulation and nonnutritive sucking showed improved neurodevelopment and achieved independent oral feeding earlier.
CONCLUSION: These findings note the potential of non-invasive interventions to enhance the development of premature infants within the NICU and to reduce stress levels in both infants and their parents. Given the profound effects that premature birth and NICU stay can have on caregivers and infants, it is important to conduct further research to evaluate other interventions aimed at alleviating stress levels.
NMMH7. The effects of childhood & adolescent stress caused by the COVID-19 pandemic on overall health
R. Glantzberg, G. Bachmann
Student at the Academy of Allied Health and Science, Rutgers University.
The COVID-19 pandemic disproportionately affected the health of kids & adolescents who underwent a host of changes that caused a tremendous deal of stress in their lives. This study aims to further understand how that stress may have impacted this population physiologically. To answer this, the database used was Google Scholar, including all articles published in the English language within the last five years. A combination of the following keywords was utilized: “COVID-19 stress”, “adolescents and teens”, “cortisol”, and “environmental stressors”. All pooled studies indicated that teens experienced long-term changes in cortisol secretion. These cortisol levels significantly increased over the course of the pandemic. Youth are especially susceptible to the impacts of chronic cortisol exposure because of their developmental status. The incongruence between biological and psychological stress could further complicate developmental concerns. The stress caused by the pandemic has been speculated to cause telomere shortening, neurophysiological disorders, epigenetic changes, precocious puberty, and obesity. One study even found that COVID-induced stress has impacted the brains of developing youth. They have been found to have reduced cortical thickness, larger hippocampal and amygdala volume, and more advanced brain age. Pediatricians and pediatric endocrinologists should be informed about the potential consequences of the exposure of stress related to the pandemic to better monitor children’s health in the coming years. Media should be involved in making an awareness campaign to inform patients and guardians as part of a global education strategy aimed at mitigating the short- and long-term effects of stress on children and adolescents.
NMMH8. Premature Newborns Risk of Development of Endocrine Disorders in Adulthood
T. Sindhu
3rd year Medical Student at Robert Wood Johnson Medical School
Premature newborns are at increased risk for disorders of the cardiac, respiratory, endocrine, metabolic, neurologic, and renal systems due to their shortened period of in-utero organ development. Specifically, regarding the endocrine system, premature dissociation of the maternal–placental–fetal unit disrupts the maintenance of physiological endocrine function in the fetus, which can have severe consequences for postnatal development. The question remains whether these early life stressors predispose premature babies to an increased risk of endocrine disorders in adulthood. A literature review was conducted using the Rutgers Libraries QuickSearch Database. Keywords included “premature”, “endocrine disorders” and “preterm”. In preterm infants, the immaturity of organ systems is associated with the insufficient secretion of hormones such as leptin, insulin, IGF-1, estrogen, and alterations in cortisol and thyroid hormone levels resulting in growth impairments, a body composition deviating from term-born infants, and unfavorable neonatal outcomes. Crump et al found that by age 18 to 44 premature infants’ risk for new-onset type 1 diabetes and lipid disorders was 1.2, 1.5 for type 2 diabetes, and significantly higher LDL and total cholesterol levels compared with full-term birth. Additional studies found preterm newborns are prone to thyroid dysfunction with an increased risk of medically treated hypothyroidism in young adulthood. Of the literature present, most agree on the increased risk of diabetes, lipid panel markers, and thyroid disorders for premature babies. However, current research is still limited on the associated risk factors during the perinatal, neonatal, and postnatal periods and the exact metabolic-endocrine disruption mechanism of premature newborns that predispose them to developing endocrine disorders in adulthood. Better understanding of these concepts could in theory guide current screening guidelines for adults born prematurely and encourage early preventive evaluation and follow-up for timely detection and treatment of endocrine disorders. Identification of protective factors that will support the long-term health trajectory and well-being of preterm-born adults can elucidate possible therapeutic interventions for premature newborns as well.
NMMH9. Caring for Patients with Trauma
J. Shinn, BS, G. Bachmann, MD, MMS
Robert Wood Johnson Medical School, Piscataway NJ, Women's Health Institute, Robert Wood Johnson Medical School, Piscataway NJ
Introduction: Adverse Childhood Experiences (ACE) have been well studied as predisposers to poorer health over a lifespan. Data suggest that individuals who have experienced more ACEs have greater incidences of STIs, maternal health problems, cancer, diabetes, heart disease, and mental illness.1As a result, these individuals often require increased usage of the healthcare system, but do not always feel comfortable in these settings given their past history of trauma.2 Our objective is to perform a reviewon best care for patients with a history of trauma using the trauma informed care model.
Methods: We conducted a systematic review using PubMed, Cochrane, and Embase databases. Inclusion criteria included manuscripts and guidelines in the English language.
Results: The Trauma Informed Care model is defined by the 4 Rs: Realize the widespread effect of trauma and understand potential paths for recovery; Recognize the signs and symptoms of trauma; Respond by fully integrating knowledge about trauma into practice; and actively resist Re-traumatization.3 Trauma informed care training in primary care physicians (PCPs) has been shown to increase patient centered care and has positive implications for better patient health and treatment compliance.4 Perhaps most essential to the model is the need to screen patients for histories of trauma. Though there is no consensus on how and when to screen, upfront universal has the ability to capture the most responses early in the physician-patient relationship.5 Care should be taken to avoid rescreening individuals as this may directly contribute to retraumatization.5 Providers should develop a therapeutic alliance with patients: this involves ensuring patient safety, clearly providing patients with personalized options and empowering them to decide their own treatment.
Conclusion: Trauma informed care is designed to address individuals with ACEs and experiences of trauma later in life which can predispose to adverse health outcomes. However, more research is needed as there are limited objective data to support specific interventions and their benefit within this model.
Steroids and Reproduction (SR)
SR1. Exposure to Cadmium Alters Placentation and Increases Macrophage Accumulation in the Placenta
B. Ames, S. Wang, D. Kozlosky, C. Gardner, D. Laskin, L. Aleksunes
Joint Graduate Program in Toxicology, Environmental and Occupational Health Sciences Institute, Department of Pharmacology and Toxicology.
The ubiquitous environmental toxicant cadmium (Cd) is known to accumulate in the placenta during pregnancy, which is associated with preterm birth and fetal growth restriction in rodents and humans. Cd is known to generate oxidative stress, recruit pro-inflammatory macrophages, and suppress macrophage responses and cytokine production. Further, alterations in macrophages responses in the placenta are associated with similar pregnancy complications such as preterm birth and pre-eclampsia. Previous work from our laboratory has demonstrated that elevated Cd concentrations in human placentas are correlated with increased villus staining for the macrophage marker CD163, but reduced staining of scavenger receptor, CD206. We hypothesize that Cd impairs the function of macrophages and alters their phenotype in the placenta. Beginning on gestational day (GD) 7, pregnant C57BL/6J dams (n=9-10/group) received either distilled water or CdCl2 (5 or 50 ppm) ad libitum through GD 17. Maternal weight gain from GD7-17 (p=0.002) and liver weight (p=0.018) were significantly decreased compared to control in the group receiving 50 ppm CdCl2. Due to the relatively low Cd exposures in this model, no change in fetal weight and length or in placental weight and area were observed. Instead, we observed a shift in placentation that favored development of the labyrinth zone in mice treated with 50 ppm CdCl2, most notably for female fetuses. Immunohistochemical staining for F4/80+ macrophages revealed a 36% and 184% increase in the labyrinth (p=0.002) and junctional zone (p=0.02), respectively, in placentas exposed to 50 ppm CdCl2. Similarly, a moderate 145% increase in the number of F4/80+ macrophages (p=0.07) was detected in the junctional zone of dams receiving 5 ppm CdCl2. No significant interactions between fetal sex were noted. Exposure of pregnant mice to low concentrations of CdCl2 during gestation alters placentation and increases the number of macrophages in the placenta.
SR2. Reproductive Health in Women: Effects of Adverse Childhood Experiences
E. Perkiss, G. Bachmann, MD, MMS
Rutgers Robert Wood Johnson Women's Health Institute, Women’s Health Institute, Obstetrics & Gynecology, Rutgers Robert Wood Johnson Medical School, New Brunswick, NJ
Introduction: Adverse childhood experiences (ACEs) are defined as potentially traumatic events occurring up to age 17. In the US, most adults experience at least one ACE; in a survey across 25 states, 61% of adults reported at least one ACE before age 18, and nearly 1 in 6 experienced at least four. A greater quantity of ACEs can potentially cause consistent activation of inflammatory pathways in response to stress, thus disrupting neuroendocrine pathways and increasing the risk for adult health problems. ACEs also have an association with heightened cortisol levels and could produce changes in brain structure and function related to emotional regulation and stress responses.
Objectives: This literature review aims to identify the effects of ACEs on women’s reproductive health.
Methods: Searching (adverse childhood experiences) AND (women’s reproductive health) on PubMed resulted in twenty-one articles. Of those, eight met the inclusion criteria of specifically addressing ACEs and reproductive outcomes in women and being written in English. One article cited two others, which also met the inclusion criteria.
Results: The most common ACEs included various types of abuse. Familial problems also were frequent adverse events. Associated reproductive outcomes were numerous and included pre-term birth, low birth weight, increased infant mortality, increased pregnancy loss, other pregnancy complications, infertility, amenorrhea, early or late menarche, increased risk of teen pregnancy, early menopause, and increased risk of oophorectomy.
Conclusions: Data suggest that ACEs may negatively affect women’s reproductive health. Future research should address the physical and psychological mechanisms through which ACEs contribute to these effects. Better understanding and prevention of ACEs should improve women’s reproductive health.
SR3. Gestational exposure to the mycoestrogen zearalenone and associations with serum sex steroid concentrations
C. Kinkade, L. Aleksunes, A. Brinker, B. Buckley, S. Growth
Environmental and Occupational Sciences Institute, Rutgers University, Piscataway, NJ, USA, Ernest Mario School of Pharmacy, Rutgers University, Piscataway, NJ, USA, Obstetrics and Gynecology, University of Rochester Medical Center, Rochester, NY, USA
Background/Aims: Zearalenone (ZEN) is a fungal-derived toxin found in global food supplies including cereal grains and processed foods. Because the chemical structure of ZEN and metabolites closely resembles 17β-estradiol (E2), they interact with estrogen receptors α (ER-α) and β (ER-β) resulting in their designation as ‘mycoestrogens’. In animal models, gestational exposure disrupts estrogen activity and impairs fetal growth. For the first time in humans, we evaluated the relationship between mycoestrogen exposure and circulating sex steroid hormones in healthy pregnant women.
Methods: Serum and urine samples were collected in each trimester from pregnant participants in the UPSIDE study (Rochester, NY, n=270). We used high performance liquid chromatography and high-resolution tandem mass spectrometry to measure total urinary mycoestrogens (ng/ml) and serum estrogens (pg/ml) (estrone (E1), estradiol (E2), estriol (E3) and androgens ng/dl (testosterone, free testosterone). Multivariable linear models were fitted for each timepoint (1st, 2nd, 3rd trimesters) using specific gravity adjusted, log-transformed mycoestrogen concentrations in relation to log-transformed sex steroids, adjusting for covariates. We additionally considered effect modification by fetal sex.
Results: ZEN and α-zearalenol (α-ZOL) were detected in >94% and >77% of participants across pregnancy (1st, 2nd, 3rd trimester medians, respectively; ZEN: 0.10, 0.12, 0.20; α-ZOL: 0.11, 0.15, 0.25 ng/ml). Models considering all pregnancies were not significant. During mid/late pregnancy, ZEN and α-ZOL were positively associated with E2 (2nd trimester: ZEN %Δ: 5.39 95%CI 0.07, 10.99, α-ZOL %Δ : 7.32, 95%CI 1.98, 12.93; 3rd trimester: ZEN %Δ : 9.37, 95%CI: 1.70, 17.62, α-ZOL %Δ : 12.15, 95%CI 4.72, 20.11) in pregnancies with female fetuses. Inverse trends between mycoestrogens and E1 and E2 were detected in pregnancies with male fetuses.
Conclusion: Mid- and late- pregnancy mycoestrogen concentrations were associated with higher E2 concentrations in pregnancies carrying female fetuses. Future work will include cord blood hormone concentrations and consideration of birth outcomes.
Supported by F31ES034269, P30ES005022, R21ES032047, T32ES019854, R01HD083369
