bluuebunny/crossref_metadata_2025_split huggingface dataset:
Functional Insights on the Human Adenine Nucleotide Translocator Interactome
Authors: Acoba Michelle Grace , Lu Ya‐Wen , Kandasamy Selvaraju , Huang Tai‐Chung , Nirujogi Raja , Pandey Akhilesh , Claypool Steven M. 
Published: November 2023

The adenine nucleotide translocator (ANT), by mediating the exchange of ADP for ATP across the inner mitochondrial membrane, lies at the core of oxidative phosphorylation (OXPHOS). Discrepancies in ANT expression and/or activity resulting to reduced energy metabolism are linked to a wide spectrum of pathologies, including lactic acidosis, progressive external opthalmoplegia, muscular dystrophy and cardiomyopathy. In yeast, the major isoform was modeled for a long time to exist in isolation, but was demonstrated to physically interact with respiratory supercomplexes (RSCs) exclusively in the context of the phospholipid cardiolipin (CL). OXPHOS in CL‐null yeast is remarkably decreased in optimal situations and completely eliminated under stressful conditions, pointing to the physiological significance of these protein‐protein associations. In this study, we determined that human ANTs also have the capacity to assemble into high molecular weight complexes. Additionally, we defined the interactome of two human ANT isoforms using an unbiased proteomics approach. Similar to the yeast orthologue, we saw RSCs and other solute carriers as binding partners, which was surprising given that the organization of yeast and mammalian ANTs is different, with Complex I missing in yeast. To probe the physiological relevance of the ANT‐RSC interaction, we deleted the major ANT isoform in HEK293, which unexpectedly only had a moderate effect on OXPHOS, indicating that the minimal levels of the other isoforms can still support respiratory capacity. As a reciprocal inquiry, we ablated OXPHOS function and found that ADP/ATP transport by ANTs is impaired. These results provide insights regarding the physiological relevance of evolutionarily conserved interactions of proteins important for mitochondrial function.Support or Funding InformationThis work was supported by the National Institutes of Health (R01HL108882) to S.M.C and pre‐doctoral fellowships from the American Heart Association (16PRE31140006 to M.G.A. and 12PRE11910004 to Y‐W.L.).
Impact of Acute Dietary Vitamin A Intake on the Measured Variable (Plasma Retinol Specific Activity) Used in Prediction of Total Body Vitamin A Stores by Retinol Isotope Dilution
Authors: Ford Jennifer Lynn , Green Joanne Balmer , Green Michael H 
Published: November 2023

Retinol isotope dilution (RID) equations are used to estimate vitamin A (VA) status (total body stores; TBS) and the efficacy of VA intervention programs after ingestion of labeled VA. Researchers have been concerned that dietary VA intake between dose administration and blood sampling (i.e., during isotope equilibration) might significantly affect TBS predictions. We recently published equations (Green et al. J Nutr 2016;146:2137–42 and 2407–11) developed from the “Olson equation” (Furr et al. Am J Clin Nutr 1989;49:713–6) that predict TBS from 1/specific activity (SAp; tracer: tracee) of plasma retinol 4 d after dosing plus several correction factors related to dose absorption and retention, equilibration of tracer between plasma and stores, and VA catabolism during equilibration; we showed that the measured variable, SAp, is the factor in the equation that has the greatest impact on TBS. To address the concern about VA intake after dosing, we used the powerful tool of model‐based compartmental analysis to simulate the effect of restricted VA intake (0 μg/d) vs. recommended intakes (400 μg/d for children and 800 μg/d for adults) on SAp at 4 d, assuming VA fractional catabolic rates of 2.2%/d (children) or 0.5%/d (adults). Intakes of 400 or 800 μg/d, compared with 0, resulted in dilution of 4 d SAp by <5%. Our results indicate that the effect of intake is small because dietary VA input represents <10% of total daily retinol input into plasma. We also further examined the contribution of 1/SAp to TBS predictions derived by model‐based compartmental analysis (WinSAAM) for data from three adult populations (Cifelli et al. J Nutr 2008;138:971–7 and Green et al. J Nutr 2016;146:2129–36). These analyses indicate that, across a wide range of TBS (35 – 2500 μmol), 1/SAp at 4 d and TBS were significantly correlated and the measured variable accounts for the vast majority of the variance in model‐determined TBS. Our results suggest that newly ingested dietary VA is a minor contributor to total daily plasma retinol input compared with VA in readily exchangeable storage pool(s); in other words, VA in exchangeable pools has a greater impact on SAp at 4 d after tracer dosing than VA intake during this period. In conclusion, these data suggest that, at VA intakes near the RDA for children and adults, dietary VA does not dilute plasma retinol specific activity at 4 d after dosing to any substantial degree and thus should have a minimal effect on this variable in the RID equation.Support or Funding InformationInternational Atomic Energy Agency, Bill and Melinda Gates Foundation (149953), and College of Health and Human Development, Penn State University
Hepatic Uptake of Vitamin A is Independent of Retinoic Acid Pretreatment in Neonatal Rats
Authors: Li Yaqi , Green Michael H. , Tan Libo , Ross A. Catharine 
Published: November 2023

Previous studies have shown that total retinoid concentration in the liver of neonatal rats would increase in response to the VARA supplementation, which is a combination of vitamin A (VA) and retinoic acid (RA). Later studies have further demonstrated that a single dose of VA may stimulate the hepatic uptake, which in turn, significantly elevated the liver total retinol concentration. In current study, we gave the neonatal rats the RA treatment prior to the VA supplementation, and examined the distinct effect of RA on VA metabolism and kinetics in the liver tissue of these young animals. Sprague‐Dawley male and female rats were mated, and the dams were fed a VA‐marginal diet (0.35 mg retinol equivalents/kg diet) and were allowed to consume food and water ad libitum. On postnatal day (P) 2 and 3, pups from experimental group received an oral dose of RA (0.625μg/g body weight), while other pups received canola oil as the control treatment. Then, an oral dose of VA (6 μg retinyl palmitate/g body weight), containing 2 μCi of [3H]‐retinol as a tracer, was administered to all pups on P4. Pups (n = 4–6/treatment/time point) were euthanized from 25 min to 30 h, a total of 12 time points, after dose administration and the liver were collected. Tissue total retinoid mass was determined by ultra‐performance liquid chromatography (UPLC), and the tissue tracer level was measured by liquid scintillation counting. Results showed that in both oil‐pretreated and RA‐pretreated groups, the fraction of retinyl ester and retinol entering the liver increased continuously and peaked at 10 h. After that, the fraction of retinol slightly declined and remained at the same level until the end of the study. However, the fraction of retinyl ester decreased gradually until the 20hr time point and showed a trend of increase afterward. At the same time, liver vitamin A content of both groups increased gradually and peaked firstly at 10 h, after a slight decrease it reached a second peak at 20 h and declined in the following period of time. In conclusion, our findings indicate that liver uptake of dietary vitamin A may be tightly controlled and would not be affected by RA supplementation.Support or Funding InformationNIH grant HD006982
Characterization of a Hepcidin Knockout Rat, a Novel Model of Iron Overload
Authors: Flores Shireen RL , Wang Xiaoyu , Ha Jungheun , Doguer Caglar , Wang Tao , Collins James F 
Published: November 2023

Genetically‐modified mouse models of altered iron homeostasis have made invaluable contributions to our understanding of the regulation of iron metabolism in humans. Emerging evidence, however, suggests that mice may not be the best model for all aspects of human physiology/pathophysiology. For example, mice display a resistance to liver cirrhosis and diabetes, which is important when investigating iron‐overload disorders, where dysfunction of the endocrine pancreas and liver damage are common consequences of excess iron deposition. Fortunately, recent advances in technology have now made genetic manipulation feasible in rats, making it possible to produce genetically‐modified rats that may better resemble some (but not all) human diseases, including those that arise from iron dysregulation.ObjectiveIn order to make comparative observations between rats and mice in regards to regulation of iron metabolism, we have developed a hepcidin knockout (Hamp−/−) Sprague‐Dawley (SD) rat. To our knowledge, this is the first attempt at direct genetic manipulation of the Hamp gene in rats. Unlike, many mouse strains used for iron biology research, SD rats are outbred, better reflecting genetic diversity in the human population.RationaleThe rationale for this experimental approach was based upon the numerous pathophysiologic differences noted between mouse models of hereditary hemochromatosis (HH) (a group of genetic iron‐overload disorders) and humans with HH.HypothesisWe hypothesized that the Hamp−/− rats would exhibit iron loading, in the liver, heart and pancreas, similar to what is seen in humans with juvenile HH (which may arise from mutations in the Hamp gene). Moreover, since rats have a longer life span than mice, we postulated that they would be more likely to develop tissue damage associated with iron loading.ResultsOur initial characterization has confirmed that the Hamp−/− rats develop early and severe multivisceral iron overload, with sparing of splenic macrophages. Interestingly, the Hamp−/− rats displayed a periportal pattern of hepatic iron deposition, similar to what is seen in humans, but distinct from the centrolobular iron‐loading pattern seen in Hamp−/− mice.ConclusionsThese data demonstrate that rats lacking hepcidin do indeed load iron, and further that key differences may exist between mice and rats in terms of iron loading patterns and regulation of iron metabolism. It is thus possible that rats may better model disturbances of iron metabolism as compared to mice, although additional studies are required to definitively make this determination.Support or Funding InformationSupported by NIH grant 1R01 DK074867 (to J.F.C.).
Effects of a high‐fat diet on spontaneous locomotor activity and blood metabolic biomarkers in Sprague Dawley rats
Authors: Chia‐Ying Wu , Yang Yan‐Ting , Chen Ching‐Han , Chang Jui‐Chou , Tsai Shiow‐Chwen , Chen Chung‐Yu , Chen Chiao‐Nan , Bernard Jeffrey R. , Liao Yi‐Hung 
Published: November 2023

PurposeObesity commonly results from the combination of excessive caloric intake and lack of physical activity. However, it is not well known if a chronic high‐fat diet leads to negative changes in resting physical activity level, which would be expected to accelerate the development of obesity. Therefore, the purpose of this study was to investigate whether a 10‐week high‐fat diet would suppress spontaneous locomotor activity in Sprague Dawley rats.MethodsSixteen male Sprague Dawley rats were randomly assigned into one of two treatment groups: 1) standard diet (SD: 10% fat, 20% protein and 70% carbohydrate) or 2) high‐fat diet (HFD: 45% fat, 20% protein and 35% carbohydrate). After one‐week acclimation, rats were fed either the SD or HFD for 10‐weeks. Throughout the dietary intervention period, body weight, water and food intake were recorded on a weekly basis. In addition, body composition (via DEXA) and spontaneous locomotor activity (via image analysis) were measured at the 9‐week time point. Immediately following the completion of the dietary intervention fasting blood glucose, plasma insulin, HbA1c and the hormones leptin and adiponectin were assess. Thereafter, an intraperitoneal glucose tolerance test (IPGTT) and insulin tolerance test (ITT) were performed at two‐day intervals in order to evaluate whole‐body glucose metabolism. All results were analyzed using a t‐Test. Significance was set at p≤0.05.ResultsSpontaneous locomotor activity (inactive time, standing frequency, movement distance, and movement velocity) were not different between SD and HFD treatment groups. Consumption of the high‐fat diet significantly increased weight gain and increased fat accumulation (whole body and visceral part), while significantly decreasing muscle mass compared to the control diet (p<0.05). In addition, rats fed the high‐fat diet showed a significant increase (by ~310%) in circulating leptin levels compared to rats fed the standard diet (p<0.05), whereas the adiponectin levels were not different between treatment groups. However, blood glucose, HbA1c and the homeostatic model assessment (HOMA) were all significantly elevated for HFD compared to SD (p<0.05). In addition, the area under the curve (AUC) for the ITT and IPGTT tests were also significantly greater for the HFD treatment group (p<0.05).ConclusionThis investigation demonstrates that despite negative changes in body composition, impaired whole‐body glycemic control, and spontaneous locomotor activity were not affected by a high‐fat diet in Sprague Dawley rats. Thus, these results suggest that excessive caloric intake may play a more critical role in the development of obesity rather than the lack of spontaneous locomotor activity.
Anti‐Obesity Effects of Solar Salt Prepared from Concentrated New Sea Water on 3T3‐L1 Adipocytes
Authors: Park Kun‐Young , Park Eui‐Seong , Ju Jaehyun , Choi Shin‐Il 
Published: November 2023

The anti‐obesity effect of Korean solar salt manufactured from concentrated new sea water on 3T3‐L1 adipocytes was studied. Our experimental groups consisted of S1 (conventionally manufactured solar salt by mixture of concentrated old‐ and new sea water), S2 (charcoal‐ and magnetic filtered during S1 preparation), S3 (solar salt made by concentrated new sea water), and S4 (charcoal‐ and magnetic filtered during S3 preparation). S3 suppressed lipid droplet formation in the differentiated adipocytes (oil‐red O staining), reduced secretion of leptin. Also, S3 significantly suppressed gene expression of lipogenesis‐ and adipogenesis‐related factors (PPARγ, SREBP‐1c, C/EBPα, LXR, LPL, FAS, DGAT and MTP; p<0.05), while promoting β‐oxidation‐related factors (PPARα and CPT‐1). Taken together, S3 was found to have anti‐obesity efficacy by suppression of lipogenesis and adipogenesis and promotion of β‐oxidation‐based lipolysis, probably owing to different processing methods to eventually lead to changes in mineral composition of the salt. Thus, a processing method of solar salt affects the anti‐obesity effect of the product.Support or Funding InformationThis work was supported by a grant 20130290 to Solar Salt Research Center of Mokpo National University from Ministry of Oceans and Fisheries of Korea.
Effects of Sulforaphane in the Tumor Microenvironment of Triple‐Negative Breast Cancer
Authors: Slattery Ed , Cline Troy , Housley Lauren 
Published: November 2023

Triple negative breast cancer (TNBC), an aggressive breast cancer phenotype that has a high rate of recurrence, is diagnosed in approximately 35,000 new breast cancer cases each year in the United States. Treatment options are limited due to the lack of common therapy targets in this subtype (estrogen receptor, progesterone receptor and human epidermal growth factor receptor 2). Since cancer progression is linked to the supportive role of healthy human cells found in the tumor microenvironment (TME), including tumor associated macrophages (TAMs), new cancer chemoprevention therapies are being sought that disrupt the recruitment and supportive activity of these tumor associated cells. There is evidence that sulforaphane (SFN), a well‐tolerated isothiocyanate derived from cruciferous vegetables, may prevent the progression of breast cancer. SFN has been shown to directly decrease cell viability in TNBC cell lines, reduce mammary tumor growth in animal models, and modify cell proliferation markers in human breast tissue. Yet, effects in human tissue have been less pronounced than in isolated cancer cells, possibly due to cell interactions in the TME. Thus, there is a critical need to determine mechanisms of SFN action in the TME of TNBC to design more effective chemoprevention and treatment strategies. TAMs, which are recruited to the TME by chemokines, are known to aid in the growth and survival of breast cancer cells. One group reported a decrease in chemokine expression in human epithelial cells following SFN treatment. The objective of this study was to establish the effect of SFN treatment on cell proliferation and chemokine secretion of human TNBC cells (MDA‐MB‐231) grown under the influence of TAMs. A conditioned‐media approach was used to model non‐contact interactions found in the TME using THP‐1 human monocytes and MDA‐MB‐231 human TNBC cells. THP‐1 cells were differentiated in 30% (v/v) cancer‐conditioned media to create TAMs. MDA‐MB‐231 cells were then grown in 30% (v/v) TAM‐conditioned media for the remaining experiments. TAM‐educated MDA‐MB‐231 were treated with SFN (10 μM) or vehicle‐control. Alterations in cell proliferation and chemokine secretion were observed in the MDA‐MB‐231 cell line after SFN treatment. This research provides evidence of the potential use of SFN in chemoprevention strategies for particularly aggressive forms of breast cancer.Support or Funding InformationCalifornia State University: 1) Student Award for Research & Creativity, 2) Research, Scholarly and Creative Activities Program
Peripheral Serotonin Deficiency May Assist Early Lactation Outcomes in Mice Fed a High‐Fat Diet
Authors: Cheng Adrienne A. , Hernandez Laura L 
Published: November 2023

Breastfeeding benefits both the mother and infant. Breastfed children have a lower incidence of type 1 and type 2 diabetes, childhood obesity, and gastroenteritis than those fed formula. Breastfeeding in the United States is much lower than ideal, with only 43.5% of mothers breastfeeding at 6 months. One of the factors is increasing prevalence of maternal obesity. As of 2009, 20.5% of women in the US are obese upon entering pregnancy. Obese women are less likely to initiate breastfeeding than their non‐obese counterparts, making their children more susceptible to health problems later in life. Decreased breastfeeding in obese women is attributed to a delay in lactation onset. Although the molecular mechanisms behind this are unknown, recent work in our lab suggests a role for peripheral serotonin deficiency in the lactating mammary gland. This study utilizes four different genotypes: wild‐type (WT), Tph1 knockout (Tph1 KO), low density lipoprotein receptor knockout (Ldlr KO), and a Tph1/Ldlr double knockout. Female mice were enrolled at 3wk and bred at 7wk. Vaginal plugs were checked daily, and the day of plug was denoted as embryonic day 0 (E0). A portion of WT and Ldlr KO mice were given their respective chow and HFD with added Tph1 inhibitor starting from E0 through their sacrifice date to assess the effect of a pharmacologic intervention in lactation outcomes. This study consists of three time points: baseline (7wk of age), lactation day 2 (L2; early lactation), and L10 (peak lactation). On L2 and L10, dams were anesthetized with isofluorane and given an i.m. injection of 0.2mL of oxytocin. Milk was collected and frozen for further protein and fatty acid analysis. Weigh‐suckle‐weigh was used to estimate total milk yield in one bout of nursing. As expected, WT and Ldlr KO mice on a HFD had increased pup mortality compared to their chow counterparts (P<0.05). At L2, these mice also exhibited difficulty lactating upon milking. However, upon the addition of the Tph1 inhibitor to the diet, WT and Ldlr KO mice have increased pup survival and restored lactation function. Previous studies in our lab have shown that mice that were fed a high fat diet (HFD) prior to and during pregnancy exhibited increased inflammatory markers in their mammary glands, similar to the inflammatory signature seen during mammary gland involution (lactation cessation). However, genetic ablation of tryptophan hydroxylase‐1 (Tph1), the rate limiting step in serotonin synthesis was able to reverse these effects. This study will further delve into how a HFD can induce inflammation in the mammary gland and whether a pharmacologic Tph1 inhibitor can be used to reverse these effects. These studies are crucial in understanding how the serotonergic system can be used clinically to improve lactation onset in obese women.Support or Funding InformationHatch funding: 142PRJ74RH
Trends in Breastfeeding in the State of Georgia, 2004–2013
Authors: Anderson Alex Kojo , Kanu Florence Adanma , Mwaura Mumbi , Ward Trina Salm 
Published: November 2023

BackgroundNational data suggest upward trends in breastfeeding initiation, exclusivity and duration across the United States but disparities between states still exist. The objective of this study is to describe trends in breastfeeding initiation and exclusivity in the state of Georgia.MethodGeorgia Pregnancy Risk Assessment Monitoring System (PRAMS) data 2004–2013 were analyzed. Participants were included in analyses if infant was born full‐term, normal weight, no birth defect, ≥ 6 months of age at time of data collection and living with the biological mother. SAS statistical software package was used for analyses.ResultsOf the 4,643 mothers included, 50.4% were White, 10.3% < 20 years, 17.9% < high school education, 60.1% married, 50.6% used Medicaid, 40.3% primiparous and 71.4% lived in an urban area. A little more than half (53.2%) reported their pregnancy was intended and 50.3% of the infants were male. 74.2% initiated breastfeeding with only 28.2% breastfeeding exclusively. There was an overall decreasing trend in breastfeeding initiation and an increasing trend for exclusive breastfeeding between 2004 and 2013 from 73.4%–70.6% and 11.9%–48.8%, respectively. Breastfeeding initiation increased only among Hispanics, 25–34 year olds, and college graduates, while exclusive breastfeeding increased among non‐Hispanic whites, Hispanics, 25–34 year olds, and across all levels of education. Despite the continued recommendations for breastfeeding by the American Academy of Pediatrics, Centers of Disease Control and Prevention and others, initiation rates in Georgia are decreasing, suggesting the need for timely education around the benefits of breastfeeding. Interestingly, there is an increasing trend for exclusive breastfeeding rate for those who initiated breastfeeding across the survey waves, which is encouraging news in the right direction.ConclusionTrends in breastfeeding initiation and exclusivity at the state level provide valuable information for state‐level public health professionals and policy makers as well as breastfeeding advocates about the challenge to meet the Healthy People 2020 objective for breastfeeding.Support or Funding InformationGeorgia Agricultural Experimental Station (HATCH # GEO00820)