bluuebunny/crossref_metadata_2025 huggingface dataset:
Competitive Enzyme Immunoassay for Vitamin B12 in Human Milk: Validation and Evaluation of Effects of Maternal Supplementation on Milk B12
Authors: Hampel Daniela , Shahab‐Ferdows Setareh , Domek Joseph M. , Siddiqua Towfida , Allen Lindsay H.
Published: June 2021

The purpose of this research was to compare and validate methods for the analysis of vitamin B12 in human milk. The Adequate Intake (AI) for infants is 0.4 μg based on an assumed milk concentration of 378pM. However, some older concentration values were determined with inaccurate and non‐validated methods. Poor maternal B12 intake or status causes low milk B12 and subsequent challenges for detection. Also, haptocorrin (HC) in human milk can produce falsely elevated or decreased values. We evaluated two assays (SimulTRAC‐SNC and IMMULITE) for their suitability for analyzing B12 in human milk. SimulTRAC‐SNB gave poor results and was highly influenced by HC, while IMMULITE can be applied without HC removal with an overall recovery rate of 78.9 % ± 9.1 (standard addition: 100 – 900 pg/mL). Diluted milk samples (24 – 193pM) were linearly correlated (r >; 0.992). Analysis of milk samples from supplemented vs. unsupplemented women respectively in Bangladesh (± 250μg/d; milk concentrations 235, 170 pM, p = 0.0389), India (± 50μg/d; 136, 87pM, p = 0.0005) and Malawi (± 2.5μg/d; 235, 185pM, n.s.) demonstrated that the method can detect increased milk B12 concentrations when supplements provide more than the RDA. However, none of the mean B12 values met the assumed milk concentration of 378pM indicating a B12 supply to the infant below the AI.

Funded by the Bill & Melinda Gates Foundation (OPP53107).
Human milk oligosaccharides modulate the duration of rotavirus‐induced diarrhea, colonic microbiota and intraluminal environment in piglets
Authors: LI MIN , WANG MEI , MONACO MARCIA H , FAHEY GEORGE C , KUHLENSCHMIDT THERESA B , KUHLENSCHMIDT MARK S , DONOVAN SHARON M
Published: June 2021

The impact of diarrheal disease and prebiotics was studied in piglets fed with formula (FF), or formula with 4 g/L human milk oligosaccharides (HMO) or prebiotics (PRE; 9:1 of short chain galactooligosaccharides and long chain fructooligosaccharides). At d10, approximately half of the piglets were infected with 5×106 focus‐forming units of group A porcine RV stain OSU. Stool consistency was monitored 3 times daily. Ascending (Asc) and descending (Dsc) colonic contents were collected at 5 d postinfection, and pH, dry matter (DM) and SCFA concentrations were measured. Asc microbiota were analyzed by 16S rRNA gene pyrosequencing. HMO and PRE groups had shorter duration of RV‐associated diarrhea than FF. RV infection increased the pH and lowered the DM of Asc and Dsc contents vs. non‐infected pigs. HMO groups had higher Dsc pH than FF and PRE, while DM of HMO and PRE groups was lower than FF. Infection increased Asc propionate concentration independent of diet. Redundancy analysis showed that microbial structure differed among infected and diet groups. Bacteroides was increased in infected pigs, and unclassified Lachnospiraceae was enriched in HMO groups. In conclusion, the intraluminal environment and microbiota were altered by RV infection and HMO supplementation. HMO and prebiotics reduced the duration of RV‐induced diarrhea, likely in part by modulating gut environment and microbiota. (Funded by R01 HD061929)

Grant Funding Source: NIH
Genetic risk scores across multiple cardiometabolic traits predict patterns of cardiometabolic risk in Filipino women
Authors: Zubair Niha , Adair Linda
Published: June 2021

We aimed to identify how genetic risk scores predicted patterns of cardiometabolic (CM) risk factors in women from the 2005 Cebu Longitudinal Health and Nutrition Survey (n = 1,492, age =36–68 y).

Previous work identified clusters of women with similar CM features: healthy, high blood pressure (BP), low HDL‐c, insulin resistant (IR), and high inflammation. Genetic risk scores for BP, LDL‐c, HDL‐c, triglycerides, glucose, and C‐reactive protein (CRP) summed the number of effect alleles known to relate to each trait. Using multinomial logistic regression, we predicted CM clusters with genetic risk scores, adjusting for population substructure, age, menopause, parity, waist circumference, BMI, and energy intake.

Compared to the healthy cluster: a higher HDL‐c genetic risk score (OR 1.12, 95% CI= 1.03–1.02) increased odds of being in the elevated BP cluster; a lower LDL‐c score (0.90, 0.82–1.00) and higher HDL‐c score (1.15, 1.08–1.23) increased the odds of being in the low HDL‐c cluster; a higher CRP score (1.28, 1.05–1.55) and higher glucose score (1.23, 1.09–1.39) increased the odds of being in the IR cluster; a higher CRP score (1.32, 1.15–1.50) and higher BP score (1.11, 1.01–1.23) increased the odds of being in the high inflammation cluster.

For most CM clusters, genetic risk across multiple traits predicted cluster membership, emphasizing the notion that many synergistic genetic effects likely influence CM risk.

Grant Funding Source: The University of North Carolina at Chapel Hill and National Institutes of Health: DK078150, HL0851445144
Effects of Agrimonia pilosa Ledeb. on insulin resistance induced in high fat diet fed rats
Authors: Lee Young Min , Nam Song Yee , Kim Young Sub , Jang Hwan Hee , Kim Sang Mi , Kim Mi Ju , Kim Haeng Ran
Published: June 2021

The purpose of this study is to investigate the effects of aqueous extract from A. pilosa on insulin resistance induced by high fat diet in rats. Using liquid chromatography‐mass spectrometry, the 3 major peaks of A. pilosa aqueous extract were identified as apigenin‐hexose, luteolin‐glucuronide, and apigenin‐glucuronide. Male Sprague Dawley rats were assigned to the following groups (n=10): normal fat diet (NF); high fat diet (HF); high fat diet with 0.1% A. pilosa aqueous extract (HFA). Experimental diets were fed for 16 weeks. In our results, high fat diet significantly increased liver and epididymal fat weight compared with NF group (p<0.05), however there was a significant decrease in liver weight by supplementation of A. pilosa aqueous extract. In oral glucose tolerance test at 15 week, we observed the blood glucose levels of the HFA group were lower than those of the HF group at 30, 60, 120 min after glucose administration (p<0.05). In addition, dietary A. pilosa aqueous extract increased serum adiponectin concentration compared with HF group (p<0.05). The mRNA expression of gluconeogenesis was decreased in HFA group (p<0.05). These data suggest that A. pilosa aqueous extract can improve insulin resistance in rats fed on high fat diet. This study was carried out with the support of the Research Program for Agricultural Science & Technology Development (Project No. PJ008554012012).
Adenovirus36 E4orf1 and Lithium inhibit GSK3, but only E4orf1 increases glucose uptake in fat cells
Authors: Dhurandhar Nikhil V , Park Soo‐Ho , Jeon Hyun‐Jung , Dubuisson Olga , Hegde Vijay , Krishnapuram Rashmi , Na Ha‐Na , Nam Jae‐Hwan
Published: June 2021

Inhibition of Glycogen synthase kinase (GSK)3 has been attempted as a therapeutic measure for diabetes, Alzheimer's disease, bipolarity, and some forms of cancers. The inhibition of GSK3 by some agents promotes cellular glucose uptake. We reported that Ad36, –a human adenovirus, increases cellular glucose uptake via its E4orf1 protein. Ad36 inhibits GSK3 by activating AKT2, a target of phosphatidyl inositol 3‐kinase activation. Here, we compared the ability of E4orf1, and Lithium (LI) ‐a widely used GSK3 inhibitor, to inhibit GSK3, and increase glucose uptake, in 3T3‐L1 cells. The, expression of E4orf1 in 3T3‐L1 preadipocytes, activated AKT, inhibited GSK3 as indicated by the phosphorylation of GSK‐3α/βserine 21 and serine 9 respectively, and increased glucose uptake, compared with cells expressing the null vector. LI (LiCl, 20mM), also inhibited GSK3, but did not activate AKT, and in fact, decreased, glucose uptake in 3T3‐L1 preadipocytes. Similarly, in 3T3‐L1 adipocytes, E4orf1 enhanced glucose uptake, whereas, LI reduced it. Thus, even if the potential of LI to inhibit GSK3 may be useful in many conditions, LI appears ineffective to improve glucose uptake in fat cells. Whereas, E4orf1 offers a template to increase glucose uptake and may be evaluated for its therapeutic application for additional diseases that would benefit from GSK3 inhibition. Funding: Vital Health Interventions.
Hypolipidemic effect of Saururus chinensis Baill in ob/ob mice
Authors: Choi Ha‐Neul , Kim Jung‐In
Published: June 2021

Tight control of hyperlipidemia is associated with the reduced risk for cardiovascular diseases in diabetic patients. The purpose of this study was to investigate the hypolipidemic effect of S. chinensis Baill in obese diabetic ob/ob mice. Four‐week‐old male ob/ob mice were fed AIN‐93G diet or diet containing a 0.3% ethanol extract of S. chinensis Baill for 10 weeks after 1 week of adaptation. Animals were sacrificed after an overnight fast. Serum triglyceride, total cholesterol, and HDL‐cholesterol levels were measured. Consumption of S. chinensis Baill significantly decreased serum triglyceride and total cholesterol levels compared with the control group (p<0.05). Serum HDL‐cholesterol levels of the S. chinensis Baill group were not significantly different from those of the control group. Non‐HDL‐cholesterol levels of the S. chinensis Baill group were significantly lower than those of the control group. These results indicate that S. chinensis Baill is effective in improving hyperlipidemia in ob/ob mice and could contribute to reducing cardiovascular complications. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2009–0090780).
Hepatoprotective effect of Saururus chinensis Baill in obese diabetic mice
Authors: Choi Ha‐Neul , Kim Ah‐Rong , Kang Min‐Jung , Kim Jung‐In
Published: June 2021

Non‐alcoholic fatty liver disease is associated with obesity, diabetes, and insulin resistance. The purpose of this study was to investigate the effect of Saururus chinensis Baill on hepatic steatosis in obese diabetic db/db mice. Four‐week‐old male db/db mice were fed AIN‐93G diet or diet containing 0.5% ethanol extract of S. chinensis Baill for 6 weeks after one week of adaptation period. Liver lipids were extracted and the amounts of triglyceride were determined. Serum monocyte chemotactic protein‐1 (MCP‐1) levels and alanine amininotransferase (ALT) activities were measured. Body weight and food intake of the S. chinensis Baill group were not significantly different from those of the control group. Hepatic triglyceride levels of S. chinensis Baill group were significantly lower than those of the control group (p<0.05). S. chinensis Baill significantly decreased serum MCP‐1 levels and ALT activities compared with the control group. These results indicate that S. chinensis Baill could alleviate hepatic steatosis in db/db mice. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2009–0090780).
LOW GLUTAMINE CONCENTRATION IS OBSERVED IN EQUINE TESTIS
Authors: Filho Helio Cordeiro Manso , da Silva Rodrigues Armele Karina , Manso Eliana C.C. , de Lima Ewerton B , Manso Helena Emilia C.C. , Wanderley Erika K. , Abreu Jose Mario Girao
Published: June 2021

Glutamine (gln) is frequently added to equine semen as a diluent to improve spermatozoa life, however there is little information about concentration of glutamine metabolism in equine testis. Gln and Glu concentrations were determined, by enzymatic analyses, in regions near the testicular vein (TA), near the external area of the testis (TB), the head of the epididymus (EH), and the tail of epididymus (ET). Samples were obtained from stallions (n=10) that underwent surgery to remove the testes. ANOVA and Tukey's test were used analyze the results, with P<5%. Tissues were washed to remove the semen present before analysis. Results showed significant diference in [glu+gln] between all four tissues. Lower [glu+gln] was observed in in EH (5.781±1.073 umol/mg of tissue) when compared with other 3 areas (ET: 10.970 ± 0.782; TA: 13.272±0.671; TB:10.840 ±1.463) (P<0.05). Overall, this work indicates considerable differences in glutamine plus glutamate concentrations in different regions of the testis. However to determine more precise the glutamine metabolism in all four tissues we need to determine GS and glutaminase activity in these tissues, and a definitive interpretation can be presented.
GLUTAMINE, BUT NOT GLUTAMATE, CONCENTRATIONS CHANGE IN EQUINE MILK DURING LACTATION
Authors: Hunka Monica Miranda , Manso‐Filho Helio CORDEIRO , DA Silva ARmele Karina Rodrigues , MELO STEphania Kartuchi , DE Lima EWerton B , Manso HELENA EMILIA C.C.C , WAnderley ERIka K. , VAZ SIMONE Gutman , D'ARAUJO JULIANA ANDRESSA
Published: June 2021

Recently it has been realized that glutamine (gln) and glutamate (glu) are the most abundant amino acids in mammalian milk. The aim of this study was to determine the gln and glu concentration in blood and milk in Quarter‐Horse mares during lactation. Nineteen (19) mares were used and samples were collected: after 8 hours parturition, and every 30 days until weaning (180 days after birth). Data were analyzed by ANOVA and Tukey's test, with P<0.05. Blood [glu] was 0.289±0.047 mol/mL at parturition day and when was 0.146+0.117 mol/ml at the end of the lactation, without significant changes (P>;0.05). In contrast, blood [gln], which was 0.582+0.078mol/mL at parturition, decreased to 0.300±0.032mol/mL at weaning (~52%). In milk, [gln] was 0.738+0.113 mol/ml 8h after delivery, increase to 1.333±0.198 mol/ml at 3 months of lactation, and then decreased to 0.511±0.098 mol/mL at weaning. Also, [glu] changes significantly, with large concentration observed at 3rd month of lactation (0.862±0.085 mol/ml) and lowest at parturition (0.449±0.057 mol/ml). These changes in blood and milk gln content indicate major changes in glutamine but the site of synthesis, skeletal muscle or mammary gland remains to be determined.
Dietary protein regulates gene expression of amino acid‐degrading enzymes through PPARα
Authors: Contreras Alejandra , Rangel Claudia , Ortiz Victor , Aleman Gabriela , Palacios‐Gonzalez Berenice , Tejero Elizabeth , Torres Nimbe , Tovar Armando
Published: June 2021

The effects of PPARα ligands, such as fatty acids, have been demonstrated in several tissues, particularly in liver, modulating the expression of enzymes for fatty acid oxidation, but recent evidence also suggest that PPARα regulates amino acid oxidation. The amino acid catabolism is regulated by hepatic amino acid‐degrading enzymes (AADE); therefore the aim of this work was to study whether PPARα regulates the expression of AADE. Male PPARα null mice were used and age‐matched with C57BL6 mice (Wild type, WT) as control group, and were fed diets containing increased amounts of dietary protein (6, 20 or 50 %). RNA was extracted from liver to assess the whole genome expression using microarrays (Mouse Gene 1.0 ST Array, Affymetrix). We found that in WT mice, the gene expression of AADE, such as Hal, Got1, Ass1, Sdh and Uroc1, was proportional to the increase in dietary protein concentration. In contrast, in PPARα null mice this effect was absent, even more, we did not observe significant differential gene expression between groups fed with 6 and 50% of protein. Moreover, some AADE such as Kynu, Oat and Acmsd, as well as the transcription factor Hnf4α showed higher gene expression in KO than WT. Network analysis suggest that PPARα regulates the expression of AADE genes through Hnf4α interaction. Thus, PPARα functions as a metabolic sensor to preserve amino acids by stimulating fatty oxidation.

Grant Funding Source: CONACYT
Branched chain amino acid plus glucose supplement reduces exercise‐induced delayed onset muscle soreness
Authors: Leahy Danielle T , Pintauro Stephen J
Published: June 2021

Supplementation with the branched chain amino acids (BCAAs) leucine, isoleucine, and valine has been used to stimulate muscle protein synthesis following exercise. The purpose of this study was to determine if BCAA plus glucose supplementation would reduce exercise‐induced delayed onset muscle soreness (DOMS). Using a double‐blind crossover design, 20 subjects (11 females, 9 males) were randomly assigned to either BCAA or placebo groups. Subjects performed a squatting exercise to elicit DOMS and rated their muscle soreness every 24 hours for four days following exercise while continuing to consume the BCAA or placebo supplement. Following a three‐week recovery period, subjects crossed‐over to the alternate BCAA or placebo treatment and repeated the same exercise and DOMS rating protocol for the next four days. BCAA supplementation resulted in an overall trend of decreased DOMS with a 33% decrease in the BCAA group versus placebo at 24 hours following exercise, however this decrease was not statistically significant (p = 0.1057). When female subjects (n=11) were analyzed separately, BCAA supplementation resulted in a significant decrease in DOMS versus placebo at 24 hours following exercise (p = 0.018). No significant effect of BCAA supplementation versus placebo was noted in male subjects.
Muscle recovery after immobilization is improved by dietary supplementation with alpha‐hydroxyisocaproic acid (HICA)
Authors: Lang Charles H. , Magne Hugues , Offord Elizabeth , Breuille Denis
Published: June 2021

We posited that αHICA, a leucine metabolite, will slow the loss or improve recovery of muscle mass in response to disuse. Adult rats were fed a diet supplemented with αHICA or an isocaloric isonitrogenous control diet. Disuse was produced by unilateral hindlimb immobilization (casting) for 7 d, using the contralateral muscle as control, or the cast removed and muscle recovered for 14 d. Casting decreased gastrocnemius mass which was associated with reduced muscle protein synthesis (MPS) and S6K1 phosphorylation, but increased proteasome activity and atrogin‐1 and MuRF1 mRNA. αHICA did not prevent casting‐induced muscle atrophy, but the typical decrement in MPS was not detected. αHICA did not alter the immobilization‐induced increase in proteasome activity and atrogene expression. After 14 d of recovery, muscle mass had returned to control values only in αHICA‐fed rats and this was associated with increased MPS and S6K1 and 4EBP1 phosphorylation in previously immobilized muscle. Proteasome activity and atrogene content remained at control levels during recovery and were not affected by αHICA. Our data suggest αHICA supplementation does not slow muscle loss produced by disuse, but does speed recovery in part by maintaining increased MPS.
The study of CELL FOOD improve cyclist aerobic endurance
Authors: Duan ligong
Published: June 2021

To investigate how CELL FOOD improve the cyclist performance.

METHODS

Part I: 10 cyclists were divided two groups randomly, CF (cell food) group and PL (placebo) group. CF group used 30 drops per time with pure water, 3 times per day without other nutritional supplements for 28 days constant. PL group used mimetic acidic water and the dose was the same. At week 0 and week 4, all subjects do graded exercises test and at week 2 and week 3, do 3 km individual time trial test at track. Part II: 11 cyclists from different team. At day 0, without cell food; at day 1, before, during and after training, all subjects drink 30 drops separately; at day 2, only after training, all subjects drink 30 drops. Part I's results show that: 1) Compare week 0 with week 4, MaxVO2, peak power, and power at 4 mmol/l lactate acid were be improved. At week 4, the same trend can be found. 2) The competition result and heart rate 2 minutes after competition of CF group are better than PL group, P <0.05. Part II result indicated that: Three parameters like live support energy, oxygen intake rate, single load time at day 1 and day 2 are better than day 0, P<0.05. CF can improve aerobic endurance by change the oxygen intake, oxygen utilization, power output of cyclist.
Post‐exercise meal timing does not influence hemoglobin synthesis in iron deficient rats
Authors: Fujii Takako , Nakashima Ayumi , Xu Minjun , Sonou Tomohiro , Matsuo Tatsuhiro , Okamura Koji
Published: June 2021

Resistance exercise increases heme synthesis in the bone marrow and the hemoglobin (Hb) concentration in iron‐deficient rats. Post‐exercise early nutrient provision facilitated exercise‐induced skeletal muscle protein hypertrophy compared to late provision in a rat study conducted for 8 weeks. However, ingesting meals early after exercise did not increase the Hb concentration in a 3‐week study using iron‐deficient rats (Fujii, T., et al. EB2012). There is a possibility that the latter study period might be too short, as no significant muscle hypertrophy was observed. Therefore, in this study, the study period was prolonged to 8 weeks to investigate the effect of post‐exercise meal timing on the Hb concentration and other blood parameters in iron‐deficient rats. Male 4‐week‐old rats were fed an iron‐deficient diet (12 mg iron/kg) and performed climbing exercise (3 d/w) for 8 weeks. The rats were divided into a group fed a post‐exercise meal early after exercise (E) or a group fed the meal 4 h after exercise (L). The skeletal muscle weights were significantly higher in the E group than in the L group. However, the Hb concentration did not differ between the groups. In addition, the bone marrow ALAD activity and plasma iron concentration did not differ. Therefore, post‐exercise meal timing that enhances exercise‐induced muscle hypertrophy has no effect on either the biosynthesis or concentration of Hb.
A Chemical Screen Reveals New Modulators of Hepcidin Expression
Authors: Fraenkel Paula G. , Volovetz Josephine , Zhen Aileen W. , Gaun Vera , Patchen Bonnie
Published: June 2021

Hepcidin, a peptide hormone produced in the liver, decreases intestinal iron absorption and macrophage iron release. To generate a tool for identifying small molecules that modulate Hepcidin expression, we stably transfected human hepatocytes (HepG2) cells with a reporter construct containing the human Hepcidin promoter upstream of a firefly reporter gene. We then used high throughput methods to screen 10,360 chemicals in duplicate for their effect on Hepcidin expression and cell viability. Regulators were identified as chemicals that caused a change >;3 standard deviations above or >;1.5 standard deviations below the mean of the other chemicals, while not adversely affecting cell viability. Using these criteria, we identified 32 small molecules that upregulated and 3 that downregulated Hepcidin expression. On retesting assays, we confirmed 22 of the initial positives (69%) and 1 of the initial negatives as regulators of Hepcidin expression. Functional classification of the positive regulators indicated: 4 anti‐inflammatory agents, 4 antimicrobials, 6 antineoplastic drugs, 4 kinase inhibitors, and 4 with other or unknown functions. We are now evaluating the mechanisms of action in human hepatocytes and in a zebrafish model. The best candidates will subsequently be tested in mouse models of iron overload syndromes. Funding: NIDDK R01 DK085250–01A1.
Risk of Oxidative Damage and Bone Resorption from Increased Iron Stores during Space Flight
Authors: Zwart Sara R. , Morgan Jennifer L.L. , Smith Scott M.
Published: June 2021

An increased amount of stored iron from neocytolysis of red blood cells and a high dietary intake of iron during space flight have raised concern about the risk of excess iron causing oxidative damage, particularly in bone. To investigate this risk, biomarkers of iron status, oxidative damage, and bone resorption during space flight were analyzed for 23 (16 M/7 F) International Space Station crewmembers. As many as 5 in‐flight blood samples and 24‐h urine pools were collected from each crewmember. Serum iron increased slightly during space flight and decreased at landing (P < 0.0004). An increase in serum ferritin early in flight (217% in women and 68% in men, P < 0.0004), returning to preflight concentrations at landing, and a decrease in transferrin and transferrin receptors during flight indicated that a transient increase in iron stores occurred. No inflammatory response was observed during flight. The oxidative damage markers 8‐hydroxy‐2′‐ deoxyguanosine and prostaglandin F2α were positively correlated (both P < 0.001) with serum ferritin. A greater area under the curve for ferritin during flight was correlated with greater changes in bone mineral density of several bone regions after flight. The results suggest that increased iron stores do constitute a risk factor for oxidative damage and bone resorption, during space flight and on Earth. Funded by the NASA Human Research Program.
Evidence for Functional Sex Dimorphism in the Serum Iron Response to Oral Iron Supplements
Authors: Orozco Monica N , Romero‐Abal María‐Eugenia , Solomons Noel W , Schümann Klaus
Published: June 2021

Background

Women appear less susceptible to some adverse effects of Fe exposure. The question remains whether this is due to their greater Fe losses and lower stores, or to other sex‐based factor(s).

Objective

To confirm previous observations suggesting sex‐associated serum Fe responses to oral Fe supplementation.

Methods

40 adults ‐‐ 10 men and 10 women with low Fe stores (<30 μg ferritin/dL) and 10 men and 10 women with adequate Fe stores (>;70 μg ferritin/dL) ‐‐ provided serum samples at baseline and at 90,180 and 270 min after ingestion of 100 mg of Fe as FeSO4. Serum Fe responses were estimated by calculating ∑ 3‐h changes in serum Fe, as determined by a ferrozine‐based assay.

Results

∑ 3‐h changes in serum Fe concentrations were 268±138, 470±212, 508±207, and 440±178 μg Fe/dL, respectively, for Fe‐sufficient men and women, and Fe‐deficient men and women. Among men, serum Fe response was significantly greater in deficiency, as expected (p=0.005). However, among women, Fe challenge produced equivalent responses, independent of Fe status (p=0.70).

Conclusion

Fe status apparently differs in its magnitude of effect on regulating the oral Fe response in women, as opposed to men, opening an opportunity for further exploration.

Funded by the Hildegard Grunow Foundation, Munich
Relationship between Manganese and Copper Intakes and Metabolic Syndrome Diagnostic Components in Korean adults
Authors: Choi Mi‐Kyeong , Bae Yun‐Jung
Published: June 2021

Recent studies have suggested that micronutrients such as magnesium, manganese and copper etc. have beneficial effects on metabolic syndrome (MetS)‐related disorders. The objective of this study was to evaluate among magnesium, manganese and copper intakes, and investigate the relation with risk of MetS among Korean adults using data from the Korea National Health and Nutrition Examination Survey in 2007–2008. A total of 3,103 adults (male: 1,140, female: 1,963) who aged 19–49 years were included in the analysis. MetS was defined using the National Cholesterol Education Program Adult Treatment Panel III criteria with a modified waist circumference cutoff (male ≥90cm; female ≥85cm). In females, subjects with high glucose had lower manganese intake than those without such abnormalities (p=0.0366). Also, among male subjects, the OR of MetS in the subjects in the highest copper intake quartile were 0.91 (95% CI: 0.46–1.80) times lower than those in the lowest quartile after adjusting for age, smoking status, alcohol consumption, physical activity and energy intake (p for trend=0.0316). However, no associations were observed between magnesium intake and MetS incidence in our subjects. These finding suggest that copper and manganese intakes may be associated with a decreased risk for MetS in the general Korean adults.
An effective approach to create subclinical vitamin K deficiency in C57BL/6 mice
Authors: Harshman Stephanie , Smith Donald , Shen Xiaohua , Booth Sarah , Fu Xueyan
Published: June 2021

The response of vitamin K (VK) in tissues subsequent to VK manipulation has not been well studied in mice, which limits effective use of genetically‐modified mouse models in VK nutritional studies. To address this concern, two studies were performed: Study #1: To determine the least amount of time required to reduce VK tissue concentrations without clinical signs of deficiency (including abnormal bleeding). Female C57BL/6 mice (n=12) were acclimated with AIN‐93 G for 1 wk and subsequently fed a VK‐deficient diet (21.4±3.0 μg phylloquinone/kg) for 0, 7, 14 and 28d while limiting coprophagy. Study #2: To determine if minimizing coprophagy makes a substantial contribution to VK tissue concentrations. C57BL/6 mice (n=64) were housed in conventional or suspended wire cages and fed AIN‐93 or VK‐deficient diet for 28d. Phylloquinone and menaquinone‐4 concentrations were measured in serum, liver, and extra‐hepatic tissues. Bleeding times were measured every 2 wks. There was a reduction of VK concentrations in all analyzed tissues within 7d (P<0.05), with the exception of body fat. VK was not detectable in liver by d28, yet there were no clinical signs of VK‐deficiency when housed in suspended wire cages. In summary, a VK deficient diet that is also menadione‐free can be used to induce low VK tissue concentrations within 7d, which will facilitate the study of VK metabolism.
Vitamin K Status and Structural Knee Osteoarthritis Characteristics in Community‐Dwelling Adults: The Healthy Aging and Body Composition Study
Authors: Shea M. Kyla , Kritchevsky Stephen B , Nevitt Michael C , Kwoh C. Kent , Booth Sarah L , Vermeer Cees , Loeser Richard F
Published: June 2021

Vitamin K dependent proteins, such as matrix gla protein (MGP), are present in joint tissues, and observational studies found osteoarthritis (OA) was prevalent in adults with lower plasma vitamin K1. We determined the cross‐sectional association between vitamin K status and presence of structural knee OA features by measuring plasma K1 and uncarboxylated MGP (ucMGP) in 408 community‐dwelling adults with qualifying knee pain who had bilateral knee MRIs (mean±SD age=74±3 y; 67% female). Analyzed at the level of the knee using logistic regression with generalized estimating equations adjusted for confounders, high plasma ucMGP (reflective of lower vitamin K status; >; the median 408pM) was significantly associated with osteophytes, bone marrow edema and subarticular cysts [respective OR(95%CIs): 1.63(1.10–2.45), 1.53(1.05–2.22), 1.74(1.21–2.51), all p<0.03]. There was a borderline association between low plasma K1 (≤ the median 0.9nM) and cartilage damage [OR(95%CI)=1.50(0.96–2.35, p=0.07]. Neither vitamin K status measure was associated with bone attrition, synovitis, or meniscal damage. Longitudinal analyses are needed to determine if vitamin K status is predictive of change in knee OA features.
Ongoing studies of novel 1‐deoxy‐sphinglipids in food
Authors: Duan Jingjing , Kelly Samuel , Sullards M. Cameron , Merrill Alfred H.
Published: June 2021

Sphingolipids have been classically defined as having sphingoid base backbones that are 1, 3‐dihydroxy, 2‐amino‐alkanes and – alkenes; however, humans and other organisms also produce sphingoid bases without the 1‐hydroxyl group (Chem Rev. 111:6387–422, 2011). These are very interesting structural variants because such compounds have been found in screens of aquatic organisms for anti‐cancer agents, and one (1‐deoxysphinganine, a.k.a. “spisulosine” and “ES‐285,” from the arctic surf clam Spisula polynyma) has shown enough promise to undergo human phase I clinical trials; and a synthetic sphingoid base analog (Enigmol) that lacks the 1‐hydroxyl‐group has anti‐tumor efficacy for colon and prostate cancer in animal models (H. Symolon et al., Mol. Cancer Ther. 10:648–57, 2011). We have developed methods for analysis of 1‐deoxy‐sphingoid bases and their N‐acyl metabolites using liquid chromatography, electrospray ionization tandem mass spectrometry and have begun to survey their occurrence in cells, tissues, and food as well as factors that influence their metabolism. These ongoing studies have found that substantial amounts (>;1 nmol/g dry weight) are present in some aquatic foods (squid and octopus), and the amounts in other meats (beef, chicken and pork) and vegetables are noteworthy. The 1‐ deoxysphingoid bases are found primarily as the N‐acyl‐derivatives rather than as the free amine, as is the case for classical sphingoid bases in most biological materials. The amounts of the 1‐deoxy‐sphingolipids are lower than “classical” sphingolipids in the foods analyzed thus far, but their relative efficacy might be comparable (or possibly greater) because 1‐deoxy‐sphingoid bases are readily absorbed because they cannot be phosphorylated and degraded in intestinal cells.

Grant Funding Source: NCI (CA137812) and NIGMS (GM069338 and GM76217)
Glucose and cyanidin‐3‐glucose interrupt quercetin metabolism in HepG2 cells
Authors: Hashimoto Naoto , Blumberg Jeffrey , Chen C‐Y. Oliver
Published: June 2021

The biological activity of flavonoids is principally exerted by their metabolites formed via phase II enzymes. The activity of these enzymes is subject to the influence of genetic, physiological, and dietary factors. We examined the effect of glucose (Glc) and the anthocyanidin cyanidin‐3‐glucose (C3G) on metabolism of the flavonol quercetin (Q) in human hepatic carcinoma HepG2 cells in vitro. Cells were treated with Glc (5.5, 30, 50 mmol/L), C3G (1, 10, 25 μmol/L) or their combination for 3 d, followed by incubation with 30 μmol/L Q for 4–24 h. Q metabolites in media and cell lysates were assessed using HPLC‐UV. Maximum production of Q metabolites by HepG2 cells treated with 5.5 mmol/L Glc was noted in cell lysate at 4 h and all intracellular Q metabolites excreted to the media at 16 h. Production of Q metabolites was inhibited in a Glc and C3G dose‐dependent manner. The combination of 50 mmol/L Glc plus 25 μmol/L C3G also decreased production of total Q metabolites due largely to the inhibitory effect of Glc. The addition of superoxide dismutase, catalase or their combination to the media containing 50 mmol/L Glc did not ameliorate the reduction in the production of Q metabolites. Thus, Q metabolism may be down‐regulated by hyperglycemia and C3G; further research is warranted to elucidate the mechanism of this action. Support by NARO and USDA.
Sulforaphane bioavailability and bioactivity in humans
Authors: Atwell Lauren L. , Clarke John D. , Hsu Anna , Bella Deborah , Stevens Jan F. , Dashwood Roderick H. , Williams David E. , Ho Emily
Published: June 2021

Sulforaphane (SFN) is an isothiocyanate derived from cruciferous vegetables with many potential health benefits. However, there is limited knowledge about its bioavailability in humans. Crucifers contain glucoraphanin (GFN), which is hydrolyzed by myrosinase to yield SFN. Myrosinase is present within the plant, and a small amount is produced by gut bacteria. We previously observed a 7‐ fold decrease in SFN bioavailability from GFN supplements with inactivated myrosinase than from fresh broccoli sprouts, suggesting that the lack of myrosinase impacts SFN bioavailability. Goals of the present study were to determine the bioavailability and bioactivity of a myrosinase‐treated broccoli sprout extract (BSE) supplement and broccoli sprouts. Participants consumed BSEs or broccoli sprouts once or twice a day (time 0 and 12h). SFN bioavailability from BSE was improved over myrosinase‐inactivated GFN supplements but was still about 3 times lower than broccoli sprouts. Consuming SFN twice daily maintained plasma metabolite levels at 24 hours, suggesting that the twice daily dosing protocol resulted in longer retention of plasma SFN levels. Nrf‐2 target genes and histone deacetylase activity were used to measure biological response to SFN treatment. This research provides important information for SFN supplement use in clinical trials. Funding: R01CA122906 & P01CA090890

Grant Funding Source: R01CA122906, P01CA090890
Internalization of intact γ‐conglutin, the lupin seed glucoselowering glycoprotein, by Hep G2 cells: biochemical and microscopy studies
Authors: Lovati Maria Rosa , Parolari Anna , Manzoni Cristina , Faoro Franco , Duranti Marcello
Published: June 2021

Gamma‐conglutin (γ‐C), a Lupinus albus seed glycoprotein, has been shown to lower blood glucose in hyperglycaemic rats and increase glucose consumption in HepG2 cells. Aim of the present work was to monitor the localization of this protein in HepG2 cells in parallel to the glucose uptake. HepG2 cells treated with γ‐C were processed to confocal microscopy and transmission electron microscopy (TEM): the protein was detected with specific antibodies. After 3 h treatment, γ‐C was accumulated inside the cells and at 6 h it was diffused as protein aggregates into the cytoplasm. At the same time, glucose uptake increases by 64% vs the control cells. At 24 h the intracellular staining was reduced and protein aggregates stacked onto the cell membranes were visible. TEM showed that microvilli were involved in trapping the protein at the beginning of the process. Microscopy observations were confirmed by 2D electrophoresis showing that the protein inside the cells consisted of its intact subunits. These findings, while confirming the remarkable glucose uptake capacity by HepG2 cells, showed that γ‐C is internalized by the cells, thus opening the way to new strategies for the understanding of the described lupin γ‐conglutin insulin‐mimetic activity.

This work was partially supported by a grant of MIUR from Italy
The effect of high dietary eicosapentaenoic acid supplementation in leukemic mice
Authors: Finch Emily , Kaushal Naveen , Kudva Avinash , Gandhi Ujjawal , Hegde Shailaja , Whelan Jay , Paulson Robert , Prabhu K. Sandeep
Published: June 2021

Current chemotherapies for treatment of chronic myelogenous leukemia (CML) do not eliminate leukemic stem cells (LSCs), leading to relapse of the disease. Discovery of alternative therapies to target LSCs is essential for elimination of the disease. We have recently reported on the endogenous production of increased levels of a novel anti‐inflammatory cyclopentenone prostaglandin Δ12‐ prostaglandin J3 (Δ12‐PGJ3), by macrophages cultured with eicosapentaenoic acid (EPA). The levels of EPA used in our studies reflect the doses prescribed by the FDA. To effectively translate these studies, we report here the endogenous production of Δ12‐PGJ3 in mice on high EPA using LC‐MS/MS. Furthermore, we report the effect of high EPA on progression of leukemia and the ability of endogenous metabolites to target the LSCs in a CML model where the hematopoietic stem cells expressing the fusion oncoprotein Bcr‐Abl are transplanted into such mice. Pathways of apoptosis activated by Δ12‐PGJ3 will be discussed.

Grant Funding Source: NIH RO1 CA 175576–01
Fructooligosaccharides augments the effect of a flavonoid, α‐ glucosyl‐isoquercitrin, on glucagon‐like peptide 1 (GLP‐1) secretion
Authors: Phuwamongkolwiwat Panchita , Hira Tohru , Hara Hiroshi
Published: June 2021

Glucagon‐like peptide‐1 (GLP‐1), which is released from the enteroendocrine cells in the presence of glucose, has shown to improve glucose tolerance. Fructooligosaccharides (FOS) and α‐ glucosyl‐isoquercitrin (Q3GM) also improve glucose metabolism but its effect on the role of GLP‐1 is unclear and warrants further investigation. We used two groups of rats (i.e., conscious and anesthetized) to conduct in vivo and in situ experiments, respectively, to investigate the secretion of GLP‐1 post administering Q3GM with‐ and without FOS. Baseline blood samples were obtained via jugular vein for in vivo but portal vein for in situ rat group with subsequent samples obtained at 15, 30, 60, 90 and 120 minutes to evaluate plasma GLP‐1 levels by using ELISA. We examined the differences among groups with 2‐way ANOVA with post‐hoc Tukey‐Kramer's test for GLP‐1 level among the group and Dunnett's for the differences from the basal blood sample. We found that for both in vivo and in situ experiments, Q3GM without FOS stimulated the secretion of GLP‐ 1, transiently. On the contrary, Q3GM with FOS significantly enhanced and prolonged plasma GLP‐1 concentrations increased by Q3GM in both in vivo and in situ experiments. Our results suggest that Q3GM with FOS has the potential for prevention or treatment of type 2 diabetes by enhancement of GLP‐1 secretion and prolongation the high plasma concentration of the antidiabetic hormone.

Grant Funding Source: Hokkaido University
Toc isomers modulation of inflammatory response in fhs 74 int fetal‐derived human intestinal cell line
Authors: Elisia Ingrid , Kitts David D.
Published: June 2021

The effect of α‐Tocopherol (Toc), γ‐Toc and δ‐Toc in modulating inflammatory response of fetal‐derived intestinal cell line (FHs 74 Int) were determined. Toc isomer‐dependently promoted interleukin (IL) 8 expression from FHs 74 Int cells, with δ‐Toc eliciting greatest pro‐inflammatory effect, followed by γ‐Toc and α‐Toc. The IL8 promoting activity was associated with non‐α‐Toc isomers ability to promote NfκB and Nrf‐2 activation as evidenced by the increased nuclear translocation of the transcription factors. Nevertheless, non‐α‐Toc isomers, particularly δ‐Toc, were found to down‐regulate the glutamate cysteine ligase (GCLC) gene expression, an Nrf‐2 target gene that is also the first and rate limiting enzyme involved in glutathione biosynthesis. The possibility that non‐α‐Toc isomers plays role in down‐regulating glutathione biosynthesis was evidenced by the finding that δ‐Toc and to a lesser extent γ‐Toc not only down‐regulated GCLC protein expression but also the total glutathione content of FHs 74 Int cells. Taken together, it is suggested that the down‐regulation of glutathione biosynthesis compromised the cellular defense system that leads to the pro‐inflammatory effect observed for the non‐α‐Toc isomers.

Grant Funding Source: Food Nutrition and Health Vitamin Research Fund
Neuroprotective effect of 1′‐acetoxychavicol acetate against amyloid β induced toxicity is related with upregulation of proteasome activity
Authors: Yaku Keisuke , Matsui‐Yuasa Isao , Yuasa‐Kojima Akiko
Published: June 2021

Introduction

1′‐Acetoxychavicol acetate (ACA) naturally obtained from the rhizomes and the seeds of the Zingiberaceae plant such as Languas galangal and Alpinia galangal exhibits anti‐carcinogenesis effect. Though Alpinia galangal has neuroprotective effect in Alzheimer disease induced mice, the effect of ACA on neuroprotection is not clear. Proteasome activity is known to decrease with aging, and the loss of proteasome activity has relation to neurodegenerative process. The present study was conducted to examine the effects of ACA on amyloid β‐protein fragment 25–35 induced neurotoxicity and proteasome activity in PC12.

Methods

The cell number of PC12 was determined by MTT assay. The proteasome activity was measured by using Proteasome‐Glo™ Chymotrypsin‐Like proteasome activity assay kit.

Results

ACA recovered the cell number which was decreased by the addition of amyloid β Fragment in PC12. The effect of ACA on amyloid toxicity was decreased by MG132 which is one of the proteasome inhibitors. ACA induced proteasome activity at early stage and remained until at least 9 hours. Then it returned to control level within 24 hours. The structure‐activity relationship of ACA on proteasome activity was different from that on anti‐carcinogenesis.

Conclusion

ACA exhibited neuroprotective effect against amyloid β‐protein fragment 25–35 induced neurotoxicity via upregulation of proteasome activity.
Synergistic Effects of Leucine and β‐Hydroxy‐β‐Methyl‐ Butyrate (HMB) with Phosphodiesterase (PDE) Inhibitors on Sirtuin Activation
Authors: Zemel Michael B , Bruckbauer Antje , Baggett Brooke
Published: June 2021

We recently demonstrated leucine and HMB synergy with resveratrol and other polyphenols to activate sirtuin signaling and downstream pathways, including mitochondrial biogenesis and fatty acid oxidation (FAO). Since resveratrol also activates Sirt1 via PDE inhibition with AMPK activation, we determined synergy between leucine or HMB and low‐dose PDE inhibitors in stimulating aerobic metabolism in adipocytes and myotubes. PDE inhibitors were studied at concentrations that exerted no independent effects in the systems studied (0.1 – 10 nM); only PDE 5 inhibitors (sildenafil and icariin) exhibited synergy with leucine (0.5 mM) or HMB (5 μM) in stimulating fat oxidation in myotubes (~200%, p=0.003) and adipocytes (~100%, p<0.05). Both PDE inhibitors exhibited comparable synergy with leucine and HMB in increasing NO production (~50%: p<0.0003) and corresponding increases in mitochondrial biogenesis (~15–30%, p<0.01). Thus, leucine‐dependent Sirt1/AMPK signaling synergizes with PDE 5‐regulated cGMP signaling to stimulate mitochondrial biogenesis and aerobic metabolism, resulting in significant effects at concentrations that are ~1% of the therapeutically‐effective plasma concentrations of these drugs.
Retention of provitamin A carotenoid in biofortified maize after processing, cooking and storage in Zambia
Authors: Mugode Luke , Kaunda Augustine , Phiri Sydney , Sikombe Thelma , Mutale Raphael , Simpungwe Eliab , Schmaelzle Samantha , Tanumihardjo Sherry , De Moura Fabiana F.
Published: June 2021

Biofortified maize hybrids with higher content of provitamin A (proA) were developed to target vitamin A deficient populations in Africa. The purpose of this study was to evaluate the degradation of proA after processing, cooking and storage among the first wave of biofortified varieties released in Zambia. The biofortified maize hybrids according to the harvesting period contained 4–11 (green maize) and 5–8 (dry maize) ìg/g of proA on a dry weight basis, respectively. The effect on the degradation of proA varied among the different genotypes. One maize genotype showed 60% retention of proA after the maize was roasted for about 20 minutes, while no degradation was observed among the other genotypes. The retention of proA ranged from 61–85% after boiling for 35 minutes at 95oC. During storage of maize grains, most of the degradation occurred in the first 15 days of storage (52–56% retention) which then stabilized retaining 30–33% of proA after 6 months of storage. In conclusion, most of the proA degradation in biofortified maize hybrids occurred during storage compared with cooking processes and the magnitude of the effect varied among the different genotypes.
Linking agriculture to nutrition – The harvest is near and how do we measure impact?
Authors: Gannon Bryan M. , Tanumihardjo Sherry A.
Published: June 2021

An emerging success story in linking agriculture to nutrition is the biofortification of staple crops with micronutrients, namely provitamin A carotenoids, iron, and zinc. As a case study, biofortification of staple crops with provitamin A carotenoids has included sweet potato, maize, and cassava using traditional breeding methods, while transgenic approaches have been utilized in Golden Rice. Human studies have included a variety of technologies, and stable isotope methods are among the most powerful to measure bioavailability and efficacy. Animal and human studies have demonstrated promising β‐carotene to retinol bioconversion factors. Long‐term intake studies in children have been completed or are underway to determine if these favorable bioconversion factors can translate into improved liver reserves of vitamin A. Using the naturally enriched, stable 13C‐signature of maize, the change in 13C‐enrichment in serum retinol was shifted in 3 to 5 year old children eating orange maize for 70 days. This shows that the β‐carotene is bioavailable and contributing to the vitamin A pool. Another study is evaluating changes in total body stores of vitamin A after 90 days of feeding orange maize to 5 to 7 year old children using the 13C2‐retinol isotope dilution test. Effectiveness studies after biofortified maize is broadly introduced are needed to investigate the feeding of β‐carotene enhanced staple crops to population groups. Biofortification of staple crops with provitamin A carotenoids is feasible and all results published to date point towards it being an effective method to improve long‐term vitamin A status of populations that adopt them.

Supported by NIH‐5T32DK007665–20 and HarvestPlus