bluuebunny/crossref_metadata_2025_split huggingface dataset:
Physiologic Role of the Menkes Copper ATPase (Atp7a) in Intestinal Iron Transport
Authors: Gulec Sukru , Ranganathan Perungavur , Lu Yan , Collins James 
Published: June 2021

Atp7a is a copper transporting ATPase involved in intestinal copper export that is strongly induced in the intestine of iron deficient rats. We evaluated the possible role of Atp7a in iron homeostasis in Atp7a knock down (KD) rat intestinal epithelial (IEC‐6) cells and Brindle mice that express a mutant, inactive form of Atp7a. In KD cells with >90% decrease in Atp7a protein levels, copper loading led to significantly increased intracellular Cu levels, demonstrating a reduction in the copper export function of Atp7a. Iron (59Fe) transport studies in IEC‐6 cells grown in bicameral cell culture inserts revealed significant transepithelial iron flux when cells were deprived of iron. Transport in Atp7a KD cells showed that 59Fe transported into cells and transferred to the basolateral chamber was reduced. Next, Brindled mice were rescued by perinatal copper injection and studies were done to consider the in vivo role of Atp7a in iron absorption. Brindled mice were anemic compared to wild type littermates, and Dmt1, TfR1 and Atp7a expression was induced by dietary iron deprivation. Furthermore, serum ferroxidase activity, reflecting ceruloplasmin, was decreased in Brindled mice as was activity of a novel cytosolic ferroxidase in enterocytes. These data demonstrate a physiologic role for Atp7a in intestinal iron transport possibly related to copper delivery to multi‐copper ferroxidases involved in iron homeostasis.Grant Funding Source : NIH
Iron transport ability of the Slc39a (ZIP) family of metal‐ion transporters
Authors: Zhang Wei , Knutson Mitchell 
Published: June 2021

Mammalian ZIP proteins represent a family of 14 transmembrane metal‐ion transporters that have been characterized mainly by their ability to transport zinc. Recently, we have found that two ZIP family members, Zip14 and Zip8, are able to mediate the cellular uptake of iron in addition to zinc. The aim of the present study was to assess the iron transport ability of the 12 other mammalian ZIP proteins. HEK 293T cells were transfected with ZIP protein‐encoding cDNAs, and iron (59Fe) uptake from ferric citrate was assessed. Expression of the ZIPs was confirmed by quantitative RT‐PCR. Among these ZIP proteins, we found that only Zip2 overexpression was able to enhance the cellular uptake of iron. However unlike iron uptake by Zip14, iron transport by Zip2 was not enhanced by the addition of L‐ascorbic acid and was not inhibited by potassium ferricyanide, indicating that iron reduction is not required prior to transport via Zip2. These studies suggest that Zip2 can transport iron, which may contribute to the altered iron homeostasis previously reported in Zip2 knockout mice (Peters et al., Genesis. 2007). Funded by NIH.Grant Funding Source : NIH
An improved method for vitamin D3 quantification in cod liver oil supplements
Authors: Scholl Chris , Metzger Brandon T. , Barnes David M. 
Published: June 2021

Sample preparation for the determination of vitamin D typically involves saponification and/or liquid/liquid solvent extraction. Cod liver oil has substantial amounts of vitamin D and has a traditional use as a vitamin D supplement, however, its oily sample matrix creates difficulty in extraction and remaining lipid residues limit UV detection. Conventional quantification requires two HPLC columns and runs (normal phase preparative and reverse phase analytical) or recently expensive mass spectrometers to measure vitamin D. Here we describe a method using normal and reverse phase SPE to achieve good cleanup of residual lipids. The complete serving size of eight commercially available cod liver oil supplements were prepared using a 2g NH2 SPE column followed by 500mg C‐18 column before analytical quantification. Using this method, a limit of detection of 20IU/serving was achieved.This method provides a simpler alternative for vitamin D detection in cod liver oil samples using SPE and a single HPLC run with UV detection and may have additional use in the quantification vitamin A and tocopherols.
Maternal DHA Supplementation Improves Lung Growth and Function in Offspring Exposed to LPS and Hyperoxia
Authors: Velten Markus , Heyob Kathryn M , Britt Rodney D , Valentine Christina J , Tipple Trent E , Rogers Lynette K 
Published: June 2021

BackgroundMaternal inflammation contributes to low birth weight and preterm delivery. Premature infants routinely receive therapeutic oxygen and exhibit persistently diminished respiratory function into adulthood.ObjectiveWe tested the hypothesis that DHA supplementation reduces the detrimental effects of a hostile perinatal environment.MethodsPregnant C3H/HeN mice were fed DHA or control diet (CD) and injected with LPS or saline on embryonic day (E) 16. Offspring were placed in room air (RA) or 85% O2 for 14 d then returned to RA. PFTs, microCTs, molecular, and histological analyses were performed.Main ResultsBirth weight was reduced in pups born to CD/LPS dams but not different than controls in the pups born to DHA/LPS dams. At 8 weeks of age, bodyweights were lower, the lungs had fewer and larger alveoli, and static compliance was decreased in mice born to dams fed CD and exposed to LPS/O2. Maternal dietary supplementation with DHA in LPS/O2 exposed mice reversed these effects even at 8 weeks with bodyweights, alveolarization, and PFT measurements in not different than CD or DHA diet, saline injected, RA exposed controls.ConclusionsThe detrimental effects of systemic maternal inflammation and neonatal hyperoxia exposure on offspring lung structure and function in adulthood can be reduced by maternal DHA supplementation.
Metabolism of Sulforaphane in Humans: Supplements vs. Whole Foods
Authors: Atwell Lauren , Clarke John D. , Hsu Anna , Allen Jeannie , Bella Deborah , Stevens Jan F. , Ho Emily 
Published: June 2021

Sulforaphane (SFN), an isothiocyanate derived from cruciferous vegetables has been studied extensively for its health benefits, including cancer prevention. Plant‐based food sources contain SFN's precursor, glucoraphanin (GFN), which must be hydrolyzed by myrosinase to yield SFN. Myrosinase is present within the plant, and a small amount is produced by certain gut bacteria. We have previously shown that subjects consuming a GFN supplement with inactivated myrosinase had significantly lower levels of SFN metabolites in plasma and urine than after consuming broccoli sprouts. These results suggest that the inactivation of the plant‐derived myrosinase results in a significant decrease in the bioavailability of SFN. The goal of the current study was to compare the bioavailability of a myrosinase‐treated broccoli sprout extract (BSE) supplement to broccoli sprouts with equivalent SFN potential. Subjects consumed either BSE or broccoli sprouts once in the morning (time 0) or twice during the day (time 0 and 12h). Plasma and urine SFN metabolites were measured using liquid chromatography tandem mass spectrometry (LC‐MS/MS) at 3, 6, 12, 24 and 48h following initial consumption. The effects of glutathione‐S‐transferase polymorphisms on SFN metabolism were also examined. This research will provide important information for using SFN in human clinical trials.Grant Funding Source: R01CA122906 & P01CA090890
Effect of phytate reduction of sorghum on zinc availability as assessed by in vitro dialysability, Caco‐2 cell uptake, and suckling rat pups
Authors: Oelofse Andre , Kruger Johanita , Du Xiaou , De Moura Fabiana F , Lönnerdal Bo 
Published: June 2021

Improved zinc availability from sorghum will benefit many malnourished communities in the least developed countries in the world, having a high prevalence of zinc deficiency. We evaluated the effect of genetic phytate reduction on zinc accessibility and uptake measured by dialysability and Caco‐2 cell uptake assays and compared those results with those from zinc absorption measured by a suckling rat pup model. Phytate reduction (80–86%) in these sorghum samples significantly increased zinc uptake and absorption. The dialysability assay was not successful in estimating zinc absorption, probably due to the nature of zinc to get adsorbed to the dialysis membrane and to precipitate. The phytate reduction resulted in the phytate:zinc molar ratio being decreased from 21.8 – 27.0 to 3.0 – 5.1, well below the critical point for inhibition of zinc uptake (10 – 15), which supports the uptake and absorption results. Caco‐2 cells can be used to estimate the zinc absorption of genetically reduced phytate sorghum.We are grateful to the Bill and Melinda Gates Grand Challenges 9, Africa Biofortified Sorghum (ABS) Project through a sub‐grant from HarvestPlus for funding this research
Changes in cell hydration and amino acid efflux related to chronically higher water intake in healthy young men
Authors: Stookey Jodi , Klein Alexis , Hamer Janice , Chi Christine , Higa Annie , Ng Vivian , Kuypers Frans , Lang Florian 
Published: June 2021

Cell hydration is integral to endocrine, immune, neural, cardiovascular, gastrointestinal, muscle, and skeletal function. Acute cell dehydration is associated with a wide variety of adverse outcomes under extreme conditions and in critical illness. Biomarkers of chronic cell hydration are needed to test for longer‐term effects under daily life conditions. In vitro, cells adapt to chronic cell dehydration by increasing protein breakdown to increase intracellular concentrations and counter the osmotic gradient. Once adapted, cells overswell if exposed to hypoosmotic conditions, and release accumulated amino acids. In vivo, in chronically stressed animals, hypoosmotic amino acid efflux is measurable in urine. This study explored for changes in cell hydration and urinary amino acid excretion in 5 healthy men (20–25y) with habitually low total water intake (<2L/d) who increased water intake to 3.5L/d over 4 wks by drinking water. Each week, urine was collected after a 750ml bolus of drinking water. Higher water intake was associated with increased urine volume, decreased urine ADH and osmolality, a downward shift in the distribution of red cell deformability over the osmolality range, and altered post‐bolus urinary amino acid excretion. More work is needed on amino acid efflux as a possible biomarker of chronic cell hydration.Funded by Danone Research, NIH #UL1 RR024131 & #5UL1RR024131‐05, and RAS #A106017.
Dose response bioavailability of green tea catechins in humans
Authors: Renouf Mathieu , Guy Philippe , Marmet Cynthia , Giuffrida Francesca , Dionisi Fabiola 
Published: June 2021

Tea is an infusion of the leaves of the Camellia sinensis plant and is the most widely consumed beverage in the world after water. Because the bioavailability of green tea catechins appears to be simpler than other polyphenols, studies on catechin bioavailability have provided precise data. However, not all is known on this topic. We investigated dose response plasma appearance of catechins after ingestion of increasing doses of commercially available infused green tea by 10 healthy volunteers.Regardless of the dose, EGC and EC were the most abundant plasmatic metabolites.Area under the curve (AUC) of EGCg and EC increased with the ingested dose of the beverage. On the other hand, the AUC of EGC and 4‐O‐Me‐EGC showed a dose response effect between the low and medium dose ingested, but saturation occurred between the medium and high doses drunk. Finally the AUC of ECg decreased with the ingested dose. The same findings were observed for Cmax. While the sum of AUC per dose showed an overall saturation effect between the medium and high dose of green tea ingested, one should be careful in interpreting the data, as the pattern of individual catechins absorption differs from one another.
Nanoencapsulation increases (−)‐epigallocatechin gallate stability and its cellular bioavailability in macrophages
Authors: Wang Shu , Zhang Jia , Sun Ming , Fan Zhaoyang 
Published: June 2021

Atherosclerotic cardiovascular disease is the leading cause of death in U.S. Macrophages are major cells responsible for atherosclerotic lesion development. (−)‐Epigallocatechin gallate (EGCG) abundant in green tea is valuable for the prevention and treatment of atherosclerosis. But low levels of stability and bioavailability limit its application in human. We encapsulated EGCG into biocompatible and biodegradable nanocarriers to increase its stability and cellular bioavailability. The stability of nanoencapsulated EGCG and native EGCG was measured at pH 7.2 at 4 °C and 25 °C. Nanoencapsulated EGCG was separated from nonencapsulated EGCG using a Sephadex™ G‐25 column. Total and nanoencapsulated EGCG was measured using a high‐performance liquid chromatography (HPLC) system. EGCG uptake by THP‐1 derived macrophages was measured using the same HPLC system. Nanoencapsulation significantly increased EGCG stability and its uptake by macrophages compared to native EGCG. This innovation portends a potential breakthrough in the prevention and treatment of atherosclerosis by using natural compounds with minimized immunogenicity and side‐effects.Grant Funding Source : NIH 1R15AT007013‐01
Validation of an in vitro digestive system for studying macronutrient decomposition in humans
Authors: Kopf-Bolanz Katrin Annika , Schwander Flurina , Gijs Martin , Vergères Guy , Portmann Reto , Egger Lotti 
Published: June 2021

In vitro digestion systems have proven to be powerful tools for understanding and monitoring the complex transformation processes that take place during digestion. In this study, human digestion was simulated with a three‐step in vitro process that was validated in depth by choosing pasteurized milk as an example of a complex food matrix. The decomposition of the macronutrients was followed over the entire digestive process to the level of intestinal enterocyte action, using protein and peptide analysis by SDS‐PAGE, reversed phase HPLC, size exclusion HPLC, and LC‐MS. The mean peptide size after in vitro digestion of pasteurized milk was 5–6 amino acids. Interestingly, mostly essential amino acids (93.6%) were released during in vitro milk digestion, a significantly different relative distribution compared to the total essential amino acid concentration of bovine milk (44.5%). All triglycerides were degraded to free fatty acids and monoacylglycerols. Herein, we present a human in vitro digestion model validated for its ability to degrade the macronutrients of dairy products comparable to physiological ranges.This project is financially supported by the Swiss federal program NanoTera.Grant Funding Source: Swiss Ferderal Research Program NanoTera
An RNA‐seq approach to identify mechanisms by which the phytochemical sulforaphane acts to prevent prostate cancer
Authors: Beaver Laura Michelle , Chang Jeff H , Williams David E , Dashwood Roderick H , Ho Emily 
Published: June 2021

Prostate cancer is one of the leading causes of cancer‐related deaths in men. A phytochemical derived from cruciferous vegetables, sulforaphane (SFN), inhibits growth and induces apoptosis in prostate cancer cells, but not normal prostate epithelial cells. SFN also inhibits histone deacetylases and DNA methyltransferases, which can epigenetically affect the expression of many genes. However, the precise mechanism by which SFN acts to target cancer cells is not well understood. We utilized high throughput RNA sequencing (RNAseq) to test the hypothesis that SFN mitigates the changes in gene expression that are critical to development of prostate cancer. Normal, early androgen‐sensitive and late androgen‐insensitive prostate epithelial cells were treated with vehicle or 15uM SFN and collected at 6 and 24h post treatment. This RNAseq approach will identify global transcriptome changes in prostate epithelial cells as they progress from normal to increasingly more aggressive cancer. Furthermore, we will determine which genes are targeted by SFN in each cell line. Taken together, this will lead to identification of genes that may be critical for the prevention of prostate cancer. This information can be leveraged to develop intervention strategies to significantly reduce the incidence and severity of prostate cancer.Grant Funding Source : Funding: P01CA090890
Retinol tends to reduce activator protein‐1 and nuclear factor kappa B mediated gene transcription in cultured human colorectal cancer cells
Authors: Olds Kaylan , Lane Michelle A 
Published: June 2021

Retinol decreases matrixmetalloproteinase 9 (MMP‐9) mRNA in colon cancer cells. The MMP‐9 promoter contains activator protein‐1 (AP‐1) and nuclear factor kappa B (NFkB) responsive elements. Our objective was to determine if retinol could affect AP‐1 and NFkB mediated gene expression. Luciferase reporter constructs containing AP‐1 or NFkB responsive elements were transfected into HCT‐116 colon cancer cells. Cells transfected with the AP‐1 construct received 20 ng/ml phorbol myristate acetate (PMA), 10 microM retinol, PMA and retinol, or vehicle (control) for 24 h. Cells transfected with the NFkB construct received 20 ng/ml tumor necrosis factor alpha (TNFalpha), 10 microM retinol, TNFalpha and retinol, or vehicle for 24 h. Treatment of cells containing the AP‐1 reporter with PMA increased luciferase activity to 280 ± 79% control. Treatment with PMA and retinol tended to reduce luciferase activity to 164 ± 39% control (P = 0.16). Treatment of cells transfected with the NFkB reporter with TNFalpha increased luciferase activity to 418 ± 142% control; while TNFalpha and retinol tended to decrease luciferase activity to 227 ± 121% control (P = 0.13). In summary, retinol tends to reduce gene transcription via AP‐1 and NFkB responsive elements. This may account, at least in part, for the ability of retinol to decrease MMP‐9 mRNA.This research was sponsored by a grant from the American Institute for Cancer Research to ML.
Expression of one‐carbon metabolism genes in cultured primary lymphocytes
Authors: Zhu Jie , Kohlmeier Martin 
Published: June 2021

The aim of the experiments was to measure gene expression in lymphocytes from peripheral blood. This procedure is to be used for assessing the impact of genomic variants on gene function.Lymphocytes were isolated from venous human blood and then cultured with phytohemagglutinin (PHA). Optimal growth was achieved with 0.8 μg/ml of purified PHA (Remel Europe, UK) resulting in about six fold expansion after 9 days. Both lower and higher PHA concentration induced proliferation less effectively. Expression of MTHFR, MTHFD1 and DHFR was measured in transcribed cDNA with validated TaqMan assays (Applied Biosystems, CA). The RNA obtained from less than one million cultured cells was sufficient for multiple expression experiments. Expression of all three targeted genes, normalized to a housekeeping gene, was approximately constant between days 6 and 10.These experiments demonstrate that gene expression analysis of lymphocytes from modest amounts of peripheral blood (a single vacutainer CPT tube with 5 ml blood) is feasible and practical.
Maternal overconsumption of calories during pregnancy programs offspring for increased adiposity through induction of adipogenic genes
Authors: potu ramesh babu , Arentson Emily J , Ragland Darryl , Donkin Shawn S , Ajuwon Kolapo M 
Published: June 2021

Obesity during pregnancy predisposes offspring to metabolic diseases such as obesity and diabetes. Few studies have explored the molecular underpinnings of the increased obesity risk in the offspring. To determine the effects of excessive maternal energy intake on adiposity in the offspring, pregnant Duroc x Landrace gilts were fed either a normal energy (mNE) or high energy diet (mHE) to induce excessive pregnancy weight gain. Offspring were sacrificed at 48h after birth and at weaning (day 21). Expression of adipogenic genes was measured using RT‐PCR. Expression of genes such as PPARγ and CEBPα was not different between mNE and mHE offspring at 48h after birth, but they were induced (P < 0.05) in mHE offspring at weaning. Additionally, SFRP4, SFRP5 and PAI‐1 were induced in mHE offspring at weaning. However, SRC1 expression was higher in mNE offspring at 48h, but not different at weaning (P < 0.05). The expression of PPARγ corepressors, SIRT1 and NCoR1, was elevated (P < 0.05) in mHE offspring at weaning. Therefore, excessive maternal energy intake during gestation programs offspring for increased adiposity through induction of adipogenic transcription factors and suppression of wnt signaling.Grant Funding Source: Showalter Research Trust
Development of diet induced obesity models in rats, comparison of metabolic and genetic effects
Authors: Hamdan-Perez Nashla , Torres Nimbe , Palacio Armando Tovar , Ordaz-Nava Guillermo , Torre-Villalvazo Ivan , Moran Sofia , Tovar-Palacio Claudia , Martinez Braulio , Hiriart Marcia , Santillan-Medina Roberto , Torres-Villalobos Gonzalo 
Published: June 2021

The development of dietary obesity models in rats mimicking human obesity may help understanding its pathology and to evaluate new therapeutic strategies for treatment. We fed male Wistar rats either with a control chow diet (C), a 45%Kcal lard diet + 5% sucrose in water (LS) a chow diet +50%Kcal lard + 5% sucrose in water (CLS) or a chow diet + 20% sucrose in water (CS) for 22 weeks (W). At the end of study LS and CS diets increased hepatic and serum triglycerides, and these differences were found in CS diet since w 4. Hepatic SREBP‐1 mRNA was higher in LS and CS but FAS mRNA only increased in CS group. The oral glucose tolerance test (OGTT) at 18 W revealed that rats fed LS, CLS and CS developed insulin resistance. Body weight was 29% higher in rats fed LS than the rest of the groups due to increased adipose tissue (AT) mass. Subcutaneous (S) and visceral (V) AT was increased 3 fold in LS with respect to C, 2 fold with respect to CLS and 75% with respect to CS. HSL, perilipin and PPAR gamma mRNA was higher in SAT of LS and CS rats, whether in VAT those genes increased only in CS and CLS. CS diet increased 3 fold the glomerular filtration rate. In conclusion this study demonstrates than the fat and carbohydrate content of the diet differentially induces obesity and comorbidities.Grant Funding Source: Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán
Genetic variation in the epithelial sodium channel (ENaC) and salt taste perception in humans
Authors: Dias Andre Gautam , El-Sohemy Ahmed 
Published: June 2021

Despite individual differences in salt taste sensitivity no studies have identified variations in genes that explain this variability. Our objective was to determine whether single nucleotide polymorphisms (SNPs) in the epithelial sodium channel (ENaC), a putative salt taste receptor with 3 subunits, modifies salt (NaCl) taste in men (n=28) and women (n=67) aged 20–35 yrs. Taste thresholds (TT) were determined using a 3 alternative forced choice staircase model with solutions ranging from 9 ×10−6 to 0.5 mol/L. Suprathreshold taste sensitivity (STT) to 5 solutions (0.01–1 mol/L) was assessed using a general Labeled Magnitude Scale. Genotypes were extracted from an Affymetrix 6.0 chip (5 SNPs in SCNN1A; 34 SNPs in SCNN1B; 13 SNPs in SCNN1G). A general linear model was used to compute differences between genotypes. No SNPs in the SCNN1A or SCNN1G genes modified salt taste. In the SCNN1B gene the rs9939129 (C>T) SNP modified TT (mmol/L ± SE) where carriers of the T allele detected significantly lower NaCl concentrations than CC homozygotes (2.33±0.76 vs 3.85±0.37, p=0.02). The rs239345 (A>T) SNP in this gene modified STT (iAUC ± SE) were those homozygous for the A allele perceived salt solutions less intensely than carriers of the T allele (70.82±12.16 vs. 96.95±3.75, p=0.02). Our findings indicate that variation in the SCNN1B gene modifies salt taste perception in humans.Grant Funding Source : The Advanced Food and Materials Network