bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of pre‐meal screen‐time activities on subjective appetite and food intake in normal weight girls
Authors: Almehmadi Amal A. , Pollard Damion , Bellissimo Nick
Published: June 2021

Evidence for an effect of pre‐meal television viewing (TVV) or other screen‐time activities on food intake (FI) is limited in children. Therefore, the purpose of this study was to determine the effect of pre‐meal screen‐time activities on subjective appetite and FI in 9–14 year old normal weight (NW) girls. NW girls (n=19) completed 45 min of TVV, computer use, video‐game playing (VGP), or sitting quietly (control). FI (mean kcal ± SEM) from an ad libitum pizza meal was measured immediately after each condition. Subjective appetite was measured at baseline, 15, 30, and 45 min. FI following TVV (575 ± 50), computer use (601 ± 42), and VGP (600 ± 54) did not differ from control (629 ± 56) (P = 0.48). However, FI was greater in girls with a higher BMI percentile (range: 65.1 – 82; n = 9) compared to the lower BMI percentile group (range: 10 – 65; n=10) (P < 0.001), but the response to screen‐time activities on FI was similar (P = 0.53). Average appetite positively correlated with FI, but only after VGP (r = 0.5; P < 0.01). Fat‐mass (kg) positively correlated with FI following TVV (r = 0.43; P = 0.06), computer use (r = 0.60; P < 0.01), and control (r = 0.59; P < 0.01). In conclusion, weight status and body fat, are stronger determinants of FI than pre‐meal screen‐time exposure in NW girls.

Supported by Danone Institute of Canada‐Grant‐in‐Aid Program.
Acute effect of avocados in meals on peptide hormones in overweight healthy adults
Authors: Haddad Ella H. , Wien Michelle , Schaffer Thomas , Oda Keiji , Sabaté Joan
Published: June 2021

Background

Avocados (Persea Americana) are a high fat, high fiber albeit low energy density food. Our goal was to examine the effect of incorporating avocados in meals on post‐meal peptide hormone concentrations.

Methods

A randomized 3×3 crossover design was used to measure peptide hormones in 26 healthy overweight (BMI 28.1±2.4 kg/m2) adults. Subjects consumed in random order a standardized breakfast meal providing 25% of their energy need (EN) on each of 3 days, followed by one of three test lunch meal: control (C) avocado‐free (35% of EN); avocado‐inclusive (AI) a meal containing avocado and matched with C for energy; and avocado‐added (AA) a meal in which a portion of avocados was added to C to provide 41% of EN. Blood samples obtained before and at 0.5 h, 1 h, 2 h and 3 h following test meals were assayed for ghrelin, leptin, gastric inhibitory peptide, glucagon‐like peptide‐1, and peptide YY by ELISA. A mixed model statistical approach was used to compare differences among test meals adjusting for time and treating participants as random effects.

Results

There was a significant difference between the 3 test meals in plasma leptin responses at 0.5 h (P<0.05) and at 3 h (P<0.0001), and an overall test meal x time interaction (P=0.05).

Conclusion

The inclusion or addition of avocado to a meal influences post meal leptin responses over a 3 hour period.

Funded by the Hass Avocado Board.
Triglyceride administration alters leptin responsiveness in rats
Authors: Vasselli Joseph R , Sanders Danielle
Published: June 2021

Leptin resistance is defined a reduced responsiveness to the feeding and body weight inhibiting effects of the hormone with increasing adiposity. One potential mechanism mediating this condition was identified by Banks et al. (Diabetes, 2004) who showed that acute iv infusion of saturated triglycerides (TG) blocks leptin transport across the blood brain barrier. To test this mechanism in vivo, we administered three equicaloric TG emulsions varying in saturation level to rats by oral gavage for 11 days. Cream (Cr), Intralipid (IL) or fish oil (FO) emulsions containing 20% lipid by weight (2.0 kcal/ml) were administered twice daily in a volume of 1.0 ml (5% of total daily calories) to groups of chow‐fed adult male S‐D rats. No change in body weight (BW) or leptin levels was seen in the groups during treatment, but FO decreased (p<0.05) and IL increased (p<0.01) circulating TG. On Day 11, rats were injected ip with 1.0 mg leptin/kg BW prior to lights out, and food intake was measured 4, 8 and 24 hrs later. Compared to intake following a saline injection given on Day 5, leptin significantly inhibited feeding at 8 and 24 hr post‐injection in the FO group (p<0.05), while the Cr and IL groups showed no change or increased feeding, respectively, at the same intervals (p<0.05). Our results show that saturation level determines responsiveness to injected leptin, and are consistent with the observations of Banks et al.

Supported by NYONRC DK‐26687.
The role of β‐carotene‐9′,10′‐oxygenase as a retinoid‐generating enzyme during mammalian embryonic development
Authors: Spiegler Elizabeth , Kim Youn-Kyung , Quadro Loredana
Published: June 2021

Vitamin A is an essential nutrient throughout the mammalian lifecycle, and is critical for proper embryonic development. β‐carotene (bC), a provitamin A carotenoid, is the major vitamin A source for most of the world. bC is converted to vitamin A by one of two enzymatic cleavage mechanisms: central cleavage by β‐carotene‐15,15′‐oxygenase (CMO1), or asymmetric cleavage by β‐carotene 9′,10′‐oxygenase (CMO2). CMO1 is thought to be more important for converting bC to vitamin A, while CMO2 functions to clear carotenoids for mitochondrial protection. Both CMO1 and CMO2 are expressed in the developing tissues, and recent work has demonstrated that CMO1 can cleave bC locally in embryos to generate vitamin A for normal development. When CMO1+/−RBP−/− dams were fed a vitamin A‐deficient diet during pregnancy, embryos were grossly malformed (eye defects, peripheral edema, and exencephaly). Supplementation of CMO1+/−RBP−/− dams with bC from 6.5–9.5 days post coitum rescued the malformations of this strain. To investigate whether CMO2 may still function as a retinoid‐generating enzyme in vivo, at least in the embryo, we fed CMOI−/−RBP−/− dams a vitamin A‐deficient diet during pregnancy and observed that bC supplementation improved the vitamin A deficiency symptoms of the double knockout embryos. We next generated triple knockout mice lacking CMO1, RBP, and CMO2. Experiments are ongoing to maintain the triple knockout dams on a vitamin A‐deficient diet throughout pregnancy and monitor the frequency of embryonic malformations, as well as maternal and embryonic vitamin A levels. These experiments will shed more light on the role of CMO2 during embryonic development.

Grant Funding Source : NIH
Serum soluble transferrin receptor concentrations in US preschool children and non‐pregnant women of childbearing age from the National Health and Nutrition Examination Survey 2003–2008
Authors: Mei Zuguo , Pfeiffer Christine M , Looker Anne C , Flores-Ayala Rafael C , Lacher David A , Mirel Lisa B , Grummer-Strawn Laurence M
Published: June 2021

Serum soluble transferrin receptor (sTfR) is elevated during increased cellular iron demands and erythropoiesis. It is recommended as a sensitive and accurate measure of iron deficiency in populations. The lack of assay standardization and of representative data on the distribution of sTfR in at‐risk populations limits its use. Using data from NHANES 2003–2008, we examined the distribution of sTfR and developed assay‐specific cutoff values for defining elevated sTfR in two US populations groups: children 1–5 y (n=2042) and non‐pregnant women 12–49 y (n=5656). On average, children had higher geometric mean sTfR concentrations (4.20 mg/L; CI: 4.15–4.26) than non‐pregnant women (3.42 mg/L; CI: 3.37–3.48) (p<0.001). Among children, those aged 1–2 y (compared to those aged 3–5 y), boys (compared to girls), and non‐Hispanic black [NHB] children (compared to non‐Hispanic white [NHW] and Mexican‐American [MA] children) had higher sTfR concentrations. Among non‐pregnant women, adolescents (12–19 y) had higher sTfR concentrations than adults aged 20–34 y but not compared to adults aged 35–49 y; NHB women (compared to NHW and MA women) and multiparous women (compared to nulliparous women) had higher sTfR concentrations. The derived cutoff values (95th percentile in a defined healthy reference population) for elevated sTfR in the US were 5.74 mg/L for children 1–5 y and 4.94 mg/L for non‐pregnant women 12–49 y.
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
Early life iron deficiency impairs spatial cognition in neonatal piglets
Authors: Rytych Jennifer L. , Elmore Monica R.P. , Dilger Ryan N. , Johnson Rodney W.
Published: June 2021

Iron deficiency is the most commonly reported nutritional deficiency in the world and has been linked to cognitive impairments in children. Our goal was to assess the impact of iron deficiency on cognition using neonatal piglets as a model for human infants, as swine neurodevelopment closely parallels human brain structure and growth. Piglets were fed one of three diets with varying concentrations of iron (Fe): control (6.3mg Fe/kg body weight [BW]), mild deficient (1.5mg Fe/kg BW), or severely deficient (0.63 mg Fe/kg BW). Piglets were placed on the diet at two days of age for four weeks. Throughout the study, hemoglobin and hematocrit levels decreased in piglets fed deficient diets. Skin pigmentation, as measured by a colorimeter in L*a*b* color space, showed reduced magneta color in the iron deficient piglets. In order to determine the effects of iron deficiency on cognition, piglets were tested in a T‐maze with extra‐maze visuospatial cues. Diet did not effect acquisition of the task; however, piglets on the severely deficient diet had significantly lower performance than controls during the reversal phase. Funding source: Internal.
Effect of dietary copper concentrations on dyslipidemia in SOD1−/− mice
Authors: Wang Shi-Kui , Roneker Carol A. , Lei Xin Gen
Published: June 2021

We previously observed dyslipidemia in Cu,Zn‐superoxide dismutase knockout mice (SOD1−/−). This study was conducted to test if the dyslipidemic phenotype in the SOD1−/− mice was ameliorated by feeding a high level of dietary Cu. Weanling male SOD1−/− mice and their wild‐type (WT) controls (3‐wk old, n = 7–10 per genotype by treatment) were fed a Torula‐yeast based, Cudeficient (< 1 mg/kg) diet (BD) or BD plus Cu (as CuSO4·5H2O) at 10 and 100 mg/kg for 6 wk. Compared with the 10 mg Cu/kg diet, the BD elevated (P < 0.05) plasma cholesterol and non‐esterifed fatty acid concentrations. However, neither the BD nor the high Cu diet (100 mg/kg) removed the genotype differences in body weight, plasma lipid profiles, or hepatic lipid profiles, although the high Cu diet decreased the relative difference in hepatic triglyceride concentration between the SOD1−/− and WT mice. Meanwhile, the BD aggravated (P < 0.05) the genotype differences in hepatic cholesterol and triglyceride concentrations. Hepatic nuclear sterol regulatory element‐binding protein‐1 and ‐2 were affected (P < 0.05) by the high Cu diet in an opposite way, but the genotype difference in these two key lipogenesis regulatory proteins remained. In conclusion, dyslipidemia in the SOD1−/− mice was not likely attributed to Cu deficiency, and could not be ameliorated by a high dietary Cu supplementation (supported in part by NIH 53018).
Supercritical fluid extraction (SFE) of vitamin D2 from UV enhanced yeast
Authors: Metzger Brandon T. , Scholl Chris , Barnes David M.
Published: June 2021

Vitamin D2, or ergocalciferol, is a fungal source of vitamin D suitable as a dietary supplement for vegans. Spent brewer's yeast is an inexpensive by‐product of the brewing industry which may be used as a starting raw material for the enrichment of D2 from the precursor ergosterol. Dried brewer's yeast was reconstituted in water and stirred during exposure to ultraviolet light (UV) for an optimized period of time to convert ergosterol to ergocalciferol. After freeze drying, the D2 enriched yeast was mixed with rice hulls and wheat germ oil for SFE extraction of D2. The SFE parameters consisted of an extraction temperature of 40°C, a solvent to feed of 75 (g CO2/g material), and a flow rate of 75 g/min in a 4L extraction vessel. 100 g of material was extracted and separated in a two stage separator (separator 1, 1500 psi, 40°C; separator 2, 750 psi, 40°C). The extraction pressure was experimentally varied (3000, 6000, and 9000 psi) to understand extraction efficiency from the yeast matrix. UV treatment of brewer's yeast led to a D2 enhancement from non‐detectable levels to 3.8 mg/g (152,000 IU/g). The percentage of D2 extracted from the yeast at the extraction pressure of 3000, 6000, and 9000 psi was 55, 35, and 31% of the starting concentration respectively. The D2 concentration was 1.5 fold higher in separator #2 compared to separator #1. SFE extraction of vitamin D2 from UV enhanced yeast is more efficient at lower extraction pressures.
Restoration of bone morphology by retinoic acid in a Vitamin A deficiency model in rats
Authors: Hamdan Hana , Standard Joseph , Herndon Betty L , Kunz Kyle , Hufstedler Jayme , Chou Joshua , Dim Daniel , Zia Hamid , Mansour Ahmad , Lankachandra Menisha , Baybutt Richard , Molteni Agostino
Published: June 2021

These studies measured the effect of vitamin A deficiency (VAD) on bone structure and reabsorption as influenced by retinoic acid or cod liver oil in a rat model. Weanling rats received a vitamin A sufficient diet (controls) or a VAD diet for 9 weeks. From week ten to thirteen the VAD group split into 4 treatment groups: continued VAD, control, supplemented with retinoic acid or with cod liver oil containing vitamins A and D. At necropsy, femurs, liver, lungs and kidneys were formalin fixed and stained for histology. Bone osteoblast counts and histopathology scoring were performed. Femur thicknesses (400×, 3 sections/bone) were recorded as internal/external diameter ratios. Femurs of VAD rats had thinner cortical bone than controls (p<0.03). Addition to the VAD diet of retinoic acid, but not of cod liver oil restored the femurs to normal thickness. VAD rats had decreased numbers of both endosteal and periosteal osteoblasts vs controls (p<0.05) which were increased significantly by retinoic acid (p<0.002). Osteoclasts were not detected. VAD diet produced lung inflammation and emphysema and increased fat in the liver. Treatment with retinoic acid or cod liver oil restored pulmonary morphology but only retinoic acid restored the liver. Kidneys were normal. Retinoic acid therefore was the most effective in reversing VAD effects.
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

Background

Maternal inflammation contributes to low birth weight and preterm delivery. Premature infants routinely receive therapeutic oxygen and exhibit persistently diminished respiratory function into adulthood.

Objective

We tested the hypothesis that DHA supplementation reduces the detrimental effects of a hostile perinatal environment.

Methods

Pregnant 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 Results

Birth 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.

Conclusions

The 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.
Over‐expression of 12/15‐lipoxygenase increases oxLDL‐induced pro‐inflammatory mediator expression and foam cell formation in rodent macrophages
Authors: Thakali Keshari M , Kang Jie , Wu Xianli
Published: June 2021

Blueberries (BB) prevent the development of atherosclerosis in apoE deficient mice, partly through suppressing 12/15‐lipoxygenase (12/15‐LOX) expression and reducing the production of oxidized lipid products. To further study the role of 12/15‐LOX in atherosclerosis and investigate the inhibitory effects of BB, a rodent macrophage cell line RAW264.7 was transfected with the 12/15‐LOX gene to investigate over‐expression of this enzyme on oxLDL‐induced inflammatory mediator production and foam cell formation. Over‐expression of 12/15‐LOX increased gene expression of the pro‐inflammatory mediators TNF‐α, RANTES and MIP in macrophage stimulated with oxLDL, which was accompanied by increased NF‐κB activity as measured by a SEAP reporter assay. When induced by oxLDL, over‐expression of 12/15‐LOX showed increased gene expression of two scavenger receptors (CD36 and SR‐A) and the reduction of two ATP‐binding cassette transporters (ABCA1 and ABCG1) expression. When assessed with Oil Red O staining, foam cell formation was increased in macrophages that over‐expressed 12/15‐LOX. Our data suggest that over‐expression of 12/15‐LOX contributes to a pro‐atherogenic phenotype. Further investigations of mechanisms underlying suppression of 12/15‐LOX by BB feeding are in progress. (USDA‐CRIS‐6251‐5100002‐06S)
Ferulic acid enhances γδ‐T cell proliferation
Authors: Rowe Cheryl A , Bonard Suzanna P , Stanilka Joy M , Percival Susan S
Published: June 2021

A previous human intervention study demonstrated that consumption of wheat bran cereal resulted in an enhancement in γδ‐T cell proliferation ex vivo. This and other studies have suggested that plant bioactive compounds can prime γδ‐T cells, thus increasing their ability to proliferate when stimulated. We asked if ferulic acid, a polyphenol found in high concentration in wheat bran, was able to promote proliferation of these cells in vitro. First, to ask about the bioavailability of ferulic acid from the wheat bran cereal, we simulated digestion in vitro and added the digest to a Caco‐2 cell monolayer. Ferulic acid in the chambers was measured by HPLC analysis. Accumulation in the basolateral chamber and decreases from the apical chamber was both time and dose dependent. Ferulic acid, as a single compound, was added to peripheral blood mononuclear cells and cultured for 24h and 10d in the presence of IL‐2, IL‐15 and PHA‐L. The presence of ferulic acid during culture resulted in an enhanced proliferation of γδ‐T cells in a dose dependent manner. We conclude that ferulic acid is bioavailable from wheat brain and can promote γδ‐T cell proliferation. Supported by Kellogg Corporate Citizen's Fund and AES, UF/IFAS
Development of a fluorometric microplate anti‐adhesion assay using uropathogenic E. coli and human uroepithelial cells
Authors: Kimble Lindsey L , Mathison Bridget D , Kaspar Kerrie L , Khoo Christina , Chew Boon P
Published: June 2021

No sensitive, high throughput fluorometric assays are available to study anti‐adhesion mechanisms of cranberry bioactives. We developed a fluorometric microplate assay to determine E. coli adhesion to human uroepithelial cells (UEC). P‐fimbriated E. coli were labeled for 30 min with 400 μM BacLight Green and pre‐incubated with urine or standard for 30 min. Fluorescent‐E. coli were added to confluent HTB‐4 UEC in mircoplates at a ratio of 400:1, incubated 1 h, washed and fluorescent intensity measured (480 abs/516 em). Specific labeling and cell adhesion were confirmed by flow cytometry. A standard curve was developed using myricetin (0–30 μg/mL) with intra‐assay CV <10% and interassay CV <15%; LLD was 0.1 μg/mL and IC50 was 0.88 μg/mL. Standard curves using quercetin and procyanidins B1, B2, and C1 showed similar inhibition. The correlation coefficient of the anti‐adhesion activity of urine samples (n = 216) from human subjects who consumed a placebo or cranberry beverage was highly significant (R2 = 0.78; P < 0.01) when this assay was compared to a 3H‐E. coli microplate assay. Anti‐adhesion activity of human urine from a pilot cranberry intervention study (n = 167) also showed good agreement with a hemagglutination assay. Therefore, we successfully developed a biologically relevant high‐throughput assay for the determination of E. coli anti‐adhesion activity in human urine. Funded by Ocean Spray Cranberries Inc.
Correlation between serum uric acid, BMI, fasting blood sugar, TG and HDL in Korean health check examinees
Authors: Ryu Hye Sook , Park Hyunjin , Yun Mieun
Published: June 2021

The purpose of this study was to assess the correlation between serum uric acid, BMI, fasting blood sugar, TG and HDL in Korean health check examinees. The respondents consisted of 2539 female adults and 3252 male adults in Gyunggi province. The subjects were divided into 4 groups by gender and serum uric acid level: FH (female hyperuricemia, n=84: 3.3% in female; whose serum uric acid level is higher than 6 mg/dl), FC (female control, n=2434: 96.7% in female; whose serum uric acid level is 6 mg/dl or under), MH (male hyperuricemia, n=529: 16.6% in male; whose serum uric acid level is higher than 7 mg/dl), and MC (male control, n=2657: 83.4% in male; whose serum uric acid level is 7 mg/dl or under). Fasting blood sugar in FH (102.5 mg/dl) was significantly higher than FC (94.0 mg/dl) (P<0.05). TG in FH (171.5 mg/dl) was significantly higher than FC (108.8 mg/dl) (P<0.001). TG in MH (196.0 mg/dl) was significantly higher than MC (155.1 mg/dl) (P<0.01). HDL‐C in MH (50.1 mg/dl) was significantly lower than MC (54.1 mg/dl) (P<0.01). TG and BMI were positively correlated with serum uric acid in female (p<0.01) and male (p<0.01) respectively. But HDL was negatively correlated with serum uric acid in female (p<0.01) and male (p<0.01). This study highlights that the serum uric acid was positively influenced fasting blood sugar, TG and BMI.
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
Variability on urinary excretion of chlorogenic acid compounds and metabolites in humans after overnight fasting and under controlled diets
Authors: Duarte Giselle , Monteiro Mariana , Marques Viviane , Farah Adriana
Published: June 2021

Large intra‐ and inter‐individual variabilities have been observed in previous studies evaluating polyphenols bioavailability in humans. The present study evaluated the intra‐ and inter‐individual variations on the urinary excretion of chlorogenic acids (CGA) and metabolites in adults (n=12), in 5 separate days, after 10h fasting and a 48h controlled phenolic free‐diet. A total of 22 compounds (359.74 ± 197.90 μmol) were identified in urine by HPLC/MS, including 6 CGA and 16 phenolic acids, varying from 1 to 10 fold for the same subject intra‐day. Only 0.36 ± 0.35 μmol of total CGA were excreted in urine, varying from 1 to 7 fold for the same subject intra‐day. A large inter‐individual variation was also observed for all compounds. The largest variation was observed for p‐coumaric acid, followed by 4‐caffeoylquinic and 3‐(4’‐hydroxyphenyl) propionic acids, whereas benzoic, hippuric and 2,4‐dihydroxybenzoic acids showed the lowest variation. The patterns of CGA and metabolites excretion were also remarkably different among subjects, but remained similar in different days for the same subjects. Concluding, the inter‐individual variability on CGA metabolism seems to be more expressive when compared to intra‐individual variability. Our results also suggest the need for long washout periods for CGA and that their metabolization routs may vary according to individuals, which should be thoroughly investigated. Financial Support: CNPQ and FAPERJ.
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
Characterization and regulation of the novel zinc transporter, ZIP11, in the murine gastrointestinal tract
Authors: Maki Alyssa B , Guthrie Gregory J , Aydemir Tolunay Beker , Cousins Robert J
Published: June 2021

ZIP11, a novel Zrt‐IRT‐like (ZIP) transporter is highly expressed in certain murine gastrointestinal (GI) tissues. We initiated research to characterize ZIP11 and to determine its regulation in a murine dietary model during low (<1 mg Zn/kg), adequate (30 mg Zn/kg) or supplemental (180 mg Zn/kg) zinc intake. qPCR & western blot analyses demonstrated that murine Zip11 expression is widely distributed among tissues. The highest relative expression of Zip11 mRNA was in the stomach, cecum, and colon of a C57BL/6 strain. ZIP11 protein was also confirmed in these tissues, along with the brain and kidney. Immunohistochemistry results revealed ZIP11 to be highly expressed in the lower region of the gastric pits, where chief cells reside and colonic epithelial cells. mRNA and protein data revealed an up‐regulation of ZIP11 in the stomach and colon during zinc deficiency. In contrast, administration of LPS reduced Zip11 expression in the stomach and had no effect in the jejunum, while Zip14 expression was increased throughout the small intestine. Our data reveals ZIP11 to be widely distributed among murine tissues, but the stomach and colon are of particular interest because of its unique tissue specificity and the response of the transporter during dietary studies. ZIP11 could be involved in the homeostatic control of zinc in mammals but may also provide tissue specific functions in the GI tract.
Association between polymorphisms in the FADS gene cluster and the plasma triacylglycerol response to an ω‐3 PUFA supplementation
Authors: Cormier Hubert , Rudkowska Iwona , Paradis Ann-Marie , Thifault Elisabeth , Garneau Véronique , Lemieux Simone , Couture Patrick , Vohl Marie-Claude
Published: June 2021

The purpose of the present study is to increase knowledge about the effects of a supplementation with omega‐3 (ω‐3) polyunsaturated fatty acids (PUFA) on lipid profile in relation to the presence of single‐nucleotide polymorphisms (SNPs) in the FADS gene cluster. Plasma lipid levels were measured in 122 subjects prior and after a 5 g/d fish oil (including 3 g/d of ω‐3 PUFA) supplement for 6‐weeks. DNA was extracted and then genotyping of 19 SNPs (covering 100% of the FADS gene cluster area) was performed using TaqMan technology. The ω‐3 PUFA supplementation had an independent effect on plasma TG levels, as expected. Thirty‐six individuals (29.6%) were classified as non‐responders, meaning there was no reduction of plasma TG levels, and eighty‐six (70.4%) were responders. Two SNPs (rs174546 and rs174611) were significantly associated (p ≤ 0.05) with plasma triglyceride (TG) levels, before and after the supplementation. These results suggest that SNPs influencing plasma TG levels are also those influencing the plasma TG response to a TG‐lowering therapy.
ZIP14 regulates tight junction protein expression in response to LPS
Authors: Guthrie Gregory J , Beker-Adeymir Tolunay , Cousins Robert J
Published: June 2021

ZIP14 is a member of the SLC39A family of zinc transporters, which uptake zinc into the cytoplasmic compartment of cells. This transporter exhibits differential tissue distribution with the intestines comprising a large relative expression level. In response to low dose lipopolysaccharide (LPS), ZIP14 expression and localization to the plasma membrane increases. In an intestinal cell model, zinc administered directly to the cells can alter the expression of tight junction (TJ) proteins. The purpose of this study is to determine if the increased ZIP14 expression during acute inflammation in the small intestines alters TJ protein expression. In vivo experiments were performed using wild type and ZIP14 ‐/‐ mice treated with intraperitoneal injections containing either saline or 2 mg/kg LPS. In vitro experiments utilized the rat intestinal cell line IEC‐6. Knockdown experiments were performed with ZIP14 siRNA on IEC‐6 cells treated with conditioned media from differentiated THP‐1 cells activated with H2O or 100 ng/mL LPS for 24 hrs. Overexpression of a ZIP14 vector for 48 hrs in IEC‐6 cells was also performed. In response to LPS stimuli, TJ protein expression was altered both in vivo and in vitro. The response of TJ proteins to LPS stimulation was altered in experimental groups lacking adequate ZIP14 expression. ZIP14 may confer a protective effect to acute LPS challenge through TJ proteins.
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.
MTHFR gene polymorphisms and circulating folate concentrations are associated with survival after breast cancer diagnosis
Authors: McEligot Archana Jaiswal , Ziogas Argyrios , Anton-Culver Hoda
Published: June 2021

We studied the influence of interactions between circulating serum folate concentrations and four polymorphic MTHFR loci [rs1476413, rs1801131 (A1298C), rs1801133 (C677T) and rs4846052] on overall survival after breast cancer diagnosis. Participants were post‐menopausal women diagnosed with breast cancer (n = 479). In the separate Cox proportional multivariate hazards models [controlled for disease stage, age at diagnosis, body mass index (BMI), parity, ER/PR status, alcohol use, vitamin use and energy intake], we found a significant inverse trend (p = 0.03; HR = 0.78, CI: 0.62 – 0.97) with increasing folate concentrations and reduced risk of dying. Also, only loci rs1801133 (C677T) was borderline significantly (p = 0.09; HR = 1.30, CI: 0.95 – 1.78) associated with survival, with the recessive allele increasing risk. The model with both circulating folate concentrations and the rs1801133 loci slightly modified the independent risk for circulating folate concentrations (p = 0.09; HR = 0.82, CI: 0.65 – 1.03) and for the rs1801133 loci (p = 0.14; HR = 1.26, CI: 0.94 – 1.73). These results suggest that serum folate concentrations at and/or after diagnosis are positively associated with survival after breast cancer diagnosis, and that the MTHFR rs1801133 loci may modify risk of dying, however larger studies are needed to more definitively assess these interactions. Supported by NCI grant K07 CA101964