bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lutein HPLC Method Comparison and Validation
Authors: Craft Neal E , Springs Donna G , Chavan Jayanthi 
Published: November 2023

Lutein is one of two pigments that accumulate in the macula of the human eye. It is yellow in color and a member of the carotenoid family. Lutein (and zeaxanthin) filter blue‐light and thereby protect the eye from damage. Lutein is very ubiquitous in plants but often masked by chlorophylls. Major dietary sources include spinach, kale, broccoli, eggs, peppers, corn and peas; however people seldom consume adequate amounts. As such, lutein has been incorporated into many dietary supplements generating a need for accurate measurement of this antioxidant pigment.ObjectivesTo compare and validate two HPLC methods for lutein analysis which use different columns and mobile phases. To compare cold and heated alkaline hydrolysis (saponification).MethodsAcceptance criteria were established as Accuracy: 90–110%; Reproducibility: within‐day <5% CV, day‐to‐day <6% CV. Both Silica and Diol HPLC columns were tested. Mobile phases were 75 hexanes/25 ethyl acetate and 96 hexanes/4 IPA, respectively. Both systems were monitored at 448 or 450nm. Saponification was necessary for samples containing lutein esters or high oil and low lutein content. Both cold and heated (56°C and 70°C) saponification methods were compared.ResultsFor both methods, calibration curves from 0.15–35 ppm were linear with r2 > 0.998. Recovery of low, mid and high lutein concentrations from spiked blank oil samples was 96–103%. Within day precision was 0.57–2.74% for esterified lutein and 0.74–3.84% for free lutein. Day‐to‐day precision was ≤3.1% for esterified lutein and ≤2.8% for free lutein. Heated saponification led to 9% less trans‐lutein and 4% less total lutein concentration compared to cold saponification.ConclusionsBoth methods met all of the criteria set forth for validation. Within day precision was less than 5% and day‐to‐day precision was less than 6%. Accuracy, as measured by spike recovery, was within the specification of 90–110%. Results were more accurate using refrigerated or 56°C saponification compared to 70°C. Both methods are aligned and produce similar results for lutein in oil‐based products. Additionally, zeaxanthin can be measured in both methods.Support or Funding InformationFunding supplied, in part, by OmniActive Health Technologies.
Low intake of fruit and vegetables, body fatness and pro‐inflammatory/pro‐oxidant status are the major determinants of plasma carotenoids reduction in free‐living adults
Authors: Manda Rodrigo Minoru , Kano Hugo Tadashi , Vilches Gabriel Nogueira , Selingardi Roberta , Abe Ligia Mayumi , Correa Camila Renata , Burini Roberto Carlos 
Published: November 2023

Previous data had shown that subjects involved in our ongoing epidemiological study “Move For Health” have a monotonous ingested (low variety) food in a poor quality (low HEI score) diet characterized by high sugar and oil servings and inadequacy of whole grains, fruits and vegetables. Fruits and vegetables are considered the main natural sources of carotenoids, detected in the plasma.The aim of the study was to investigate the relationship of food intake with major circulating carotenoids, and related components either plasma lipids and/or pro‐inflammatory, pro‐oxidative markers, in female adult subjects enrolled in a lifestyle modifications Program (LSMP).The cross‐sectional analysis involved retrospective data of demographic, anthropometric, dietary (24h food intake recall), plasma general chemistry (lipids), oxidative inflammatory (hs‐CRP, γ‐GT, HOMA‐IR, MDA, uric acid) markers and lipo‐soluble vitamins (retinol, α‐tocopherol, β‐carotene and others carotenoids (β‐criptoxanthin, lutein and lycopene). Mean (±SD) were compared by Student t‐test for age, and continued values among variables were correlated by the Spearman test for p=0.05.Overweight (29.6±5.74 kg/m2) high body fat (37.0±8.79%) and abdominal fat (WC=96.8±13.8cm) women, presenting high plasma total cholesterol (tchol=201±30.1mg/dL) and triglycerides (TG=154±49mg/dL) levels along with high HOMA‐IR (3.38±3.05), presented circulating vitamins inside their normal range. However, the ingested sources of these vitamins were considered below the recommended amounts. All but lycopene carotenoids plasma levels were negatively related to BMI, %fat and WC as well as plasma TG (including lycopene), tchol and LDL‐chol (only β‐carotene). The inflammatory/oxidative/insulin resistance‐related markers showed reverse relationship with plasma carotenoids, more complete with γ‐GT and MDA (for β‐carotene, β‐criptoxanthin and lutein). The best relationship (β‐carotene, β‐criptoxanthin and lutein) was found with the food intake of fibers and vegetables. Thus, free‐living adults presenting lower intake of carotenoids might affect their plasma levels, which is aggravated by their hyperadiposity pro‐inflammatory status. Nutritional intervention with fruits and vegetables adequacy would reverse or attenuate the situation.Support or Funding InformationSupported by CAPES and CNPq
The physical stability comparison of two types of resveratrol nanocarriers
Authors: Zu Yujiao , Wang Shu 
Published: November 2023

ObjectiveResveratrol (R) has anti‐obesity bioactivities, but its low level of aqueous solubility and stability limits its application. We successfully synthesized biocompatible and biodegradable R encapsulated lipid nanoparticles (R‐nano) and R encapsulated liposomes (R‐lipo). The objectives of this study are to measure the characteristics of R‐nano and R‐lipo and to compare their physical and chemical stability at three different temperatures.MethodThe particle size and polydispersity index (PI) of R‐nano and R‐lipo were measured using a Brookhaven BI‐MAS particle size analyzer, and their zeta potential were measured using a Brookhaven ZetaPALS analyzer. The R‐nano and R‐lipo were stored in dark at 4°C, 22°C and 37°C for 7 days, and their physical and chemical stability was measured every 24 hours. The freshly made R‐nano and R‐lipo were aliquot into black tubes and stored at the above three temperatures. The mean particle size, PI and zeta potential were measured every 2 hours for the first 10 hours, and every 24 hours for 7 days. The chemical stability of R‐nano, R‐lipo and native R were measured by a high‐performance liquid chromatography system.ResultsThe mean particle size of freshly made R‐nano and R‐lipo were around 140 nm and 110 nm, respectively. The PI values of both R‐nano and R‐lipo were less than 0.3. Moreover, the zeta potential of R‐nano and R‐lipo were around −19 and −28mV. As compared to native R, R‐nano and R‐lipo dramatically increased the aqueous solubility of R by more than 30 times. Nanoencapsulation also significantly enhanced the chemical stability of R. The diameter and zeta potential of R‐lipo did not changed significantly at all temperatures (4°C, 22°C and 37°C) for 7 days, and the PI of R‐lipo remained under 0.3. R‐nano had lower physical stability than R‐lipo at 22°C and 37°C. After incubating at 22°C for 24 hours and at 37°C for 8 hours, the particle size was dramatically increased, and their PI and zeta potential values were also increased. The particle size, PI and zeta potential of R‐nano were stable at 4°C for 24 hours.ConclusionsBoth R‐lipo and R‐nano increased the aqueous solubility and stability of R. R‐lipo had higher physical stability than R‐nano. We are investigating the anti‐obesity bioactivities of nanoencapsulated R.Support or Funding InformationGrant Funding Source: NIH 1R15AT008733‐01
Is Plasma Plant Sterols a Biomarker of Plant Sterols Intake Estimated from FFQ and Multiple Dietary Recalls?
Authors: Sirirat Rawiwan , Haddad Ella , Mashchak Andrew , Jaceldo‐Siegl Karen 
Published: November 2023

BackgroundPlant sterols have many health benefits including reduce risk of heart diseases and prevent cancers. We have previously found that plasma plant sterols are higher in vegans than non vegetarians. It is not clear, howeer, if plant sterols in plasma reflect dietary intake.ObjectiveWe compared plasma plant sterol levels with plant sterol intake estimated from food frequency questionnaire (FFQ) and 24‐hours dietary recalls (24‐HR) of the Adventist Health Study ‐2 cohort.DesignPlasma sitosterol and campesterol were measured by gas‐liquid chromatography. Intake of β‐sitosterol and campesterol was estimated from FFQ and six 24‐HR. The plant sterols content of foods were derived primarily from USDA SR 27 and also from available literature between years 1999–2011.ResultsMean (SD) age of the analytic cohort was 60 (14) years, and included 34% men, and 54% non‐Blacks. Mean (SD) was 4.2 (2.3) μg/ML for plasma campesterol and 3.3 (1.6) μg/ML for plasma sitosterol. In FFQ, mean (SD) intake of campesterol and β‐sitosterol in mg/d was 38.03 (22.06) and 170.57 (95.72), respectively. Mean (SD) intake of campesterol and β‐sitosterol from 24‐HR were 22.80 (13.82) mg/d and 91.70 (50.09) mg/d, respectively. Correlations between plasma concentration and 24‐HR for campesterol and sitosterol were not significant. The correlation between 24‐HR and FFQ was 0.16 (p <.0001) for campesterol and 0.24 (p <.0001) for sitosterol.ConclusionPlasma concentration of plant sterols does not appear to be a biomarker of phytosterols intake. Although weak, correlations between plant sterols intake in FFQ and 24‐HR may be improved with adjustment for measurement errors or by food group assessment.Support or Funding InformationThis project was funded by Unilever. Collection of blood and dietary recalls was funded by the National Cancer Institute.
Nutritional Polyphenol Metabolites’ Kinetics in Plasma, Urine, and Saliva Following Consumption by Normal, Healthy, Subjects
Authors: Choy Ying Yng , Hester Shelly N. , Mastaloudis Angela , Wood Steve M. , Gray Russell , O'Donnell Robert P. , Barger Jamie L. , Quifer‐Rada Paola , Lamuela‐Raventós Rosa M. , Waterhouse Andrew L. 
Published: November 2023

Several studies point to the health benefits of dietary polyphenols yet limited data can be found on the metabolites found in plasma and urine and negligible on saliva from supplemented polyphenol ingredients. Therefore this study was completed to obtain a better understanding of the kinetics of polyphenol ingredients in three fluids (plasma, urine, and saliva). A novel, dietary nutritional supplement containing several polyphenols including resveratrol, quercetin, anthocyanins, heperitin, naringin, d‐limonene combined with fish oil, vitamins, and carotenoids was studied. Healthy subjects (n=6 males and n=6 females) were fed three different acute doses in this cross‐over design, one treatment at a time, with at least a one week washout interval. Blood plasma, saliva and urine samples were collected for metabolite analysis at baseline, 0.5, 1, 2, 3 (only plasma and saliva), 8, and 24 hours post intake. Blood was also collected in PAXgene tubes at baseline, 8, and 24 hours post intake for gene expression analysis. Metabolites analyzed included 3‐hydroxyphenylacetic acid, 4‐hydroxyhippuric acid, hippuric acid, hesperetin, naringenin, naringenin 7‐O‐glucuronide, protocatechuic acid, quercetin, quercetin 3‐O‐glucuronide, resveratrol 3‐O‐glucuronide, resveratrol 4‐O‐glucuronide, trans‐resveratrol, and cis‐resveratrol. Most of the metabolites that were analyzed peaked at 2 and 8 hours and returned to baseline by 24 hours in plasma, urine, and saliva. A dose response was not observed for all metabolites in plasma, urine, and saliva. Female and male kinetics of many of the polyphenols and metabolites appears to be different. This data is important in further understanding the levels of these polyphenols and their metabolites, so that tissue exposure can begin to be assessed.Support or Funding InformationSupported by Nu Skin Enterprises, Provo, Utah
Curcumin metabolites induce autophagic cell death through PI3K/AKT‐mTOR signaling in COLO205 human colon adenocarcinoma cells
Authors: Cheng An‐Chin , Lee Ming‐Fen , Chiou Bin‐Hao , Yen Yu‐Zhu , Pan Min‐Hsiung 
Published: November 2023

Autophagy, type II programmed cell death, is crucial for maintaining cellular homeostasis and plays a role in many diseases including cancer. Tetrahydrocurcumin (THC), a major metabolite of curcumin, has been demonstrated an autophagy‐inducing effect in human HL‐60 promyelocytic leukemia cells. In the present study, we found that THC and two other curcumin metabolites, tetrahydrodemethoxycurcumin (THDC) and tetrahydrobisdemethoxycurcumin (THBC), decreased the survival ratio of COLO205 human colon adenocarcinoma cells, but did not increase the sub‐G1 cell population, one of the hallmarks of apoptosis. THC and its derivatives, THDC and THBC, induced autophagic cell death in COLO205 cells by increasing acidic vascular organelle (AVO) formation, an autophagy marker. The results also indicated that the numbers of the methoxy groups in the curcumim metabolites were negatively correlated with their autophagy‐inducing effects. The amounts of LC3‐I/II and beclin‐1 increased and the phosphorylation levels of PI3K, AKT, and mTOR decreased with the addition of THBC in a dose‐dependent manner. Taken together, these data demonstrated the anticancer efficacy of curcumin metabolites which suggest a potential translational application for the prevention of human colon cancer.Support or Funding InformationThis study was supported by NSC102‐2320‐B‐309‐001‐MY3 to M‐F Lee and NSC100‐2313‐B‐309‐003 to A‐C Cheng.
Protective effect of betaine against liver steatosis involves depletion of homocysteine via upregulation of betaine‐homocysteine methyltransferase (BHMT)
Authors: Ahn Chul W , Jun Doo S , Na Jong D , Choi Yeo J , Kim Young C 
Published: November 2023

Accumulation of lipids in hepatocytes plays an important role in the development of metabolic diseases. Adenosine monophosphate‐activated protein kinase (AMPK) has emerged as a potential target for intervention in hepatic fat accumulation. In this study we determined the effect of betaine, a substrate utilized in remethylation of homocysteine to methionine in the transsulfuration reactions, on the AMPK pathway and lipogenesis in liver. Male rats were fed methionine‐choline deficient diet (MCD) for 3 wk. Intake of MCD resulted in a marked accumulation of lipids in liver. Phosphorylation of AMPK, acetyl‐CoA carboxylase (ACC), sterol regulatory element binding protein (SREBP)‐1c, and liver kinase B1 (LKB1) was inhibited significantly. Betaine supplementation (1 %) prevented or reversed all the changes induced by MCD intake. The underlying mechanism of AMPK activation provided by betaine was investigated using different hepatoma cell lines. Betaine treatment enhanced phosphorylation of AMPK, SREBP‐1c, and ACC in H4IIE, but not in HepG2 cells. Betaine‐homocysteine methyltransferase (BHMT) was induced and homocysteine levels were reduced by betaine only in H4IIE cells. Homocysteine treatment decreased pAMPK, pACC, pSREBP‐1c, and pLKB1 in a dose‐dependent manner, which was all prevented by betaine. BHMT protein expression was inhibited by homocysteine and betaine reversed its inhibition in H4IIE cells. Treatment of hydroxylamine, a cystathionine β‐synthase inhibitor, resulted in a significant increment of homocysteine, while decreasing phosphorylation of AMPK, ACC, and SREBP‐1c. Metformin treatment to HepG2 cells increased phosphorylation of AMPK, ACC, and SREBP‐1c, which was suppressed by homocysteine. Taken together, the results show that the elevation of homocysteine in hepatocytes may be responsible for the inactivation of the AMPK pathway. It is strongly suggested that anti‐lipogenic activity of betaine is associated with depletion of homocysteine via upregulation of BHMT.Support or Funding InformationSupported by National Research Foundation (NRF) grants (No. 2014‐R1A2A1A11052967 and No. 2009‐0083533) funded by the Ministry of Education, Science and Technology (MEST), Korea.
Role of acetyl‐CoA metabolizing enzyme ACSS2 and dietary acetyl‐CoA precursors in transcription and lipid disorder
Authors: Moreau Regis , Erickson Anjeza , Bruner Noel , Xia Mengna , He Bo 
Published: November 2023

Dyslipidemia and fatty liver that occur secondarily to obesity result from extensive gene expression changes in tissues throughout the body. By converting acetate into acetyl‐CoA, acetyl‐CoA synthetase 2 (ACSS2, cytoplasmic) raises the levels of acetyl‐CoA with an expected impact on histone acetylation and lipid production. We created two stable AML12 liver cell lines that exhibit normal or high (mRNA, +1300%; protein, +1600%) levels of ACSS2 and in this model asked how dietary precursors of acetyl‐CoA modulate histone acetylation and lipid metabolism. ACSS2 raised global histone H3 acetylation (+53%), increased fatty acid synthase abundance (+67%), and induced albumin expression (cellular, +643%; secreted, +284%). Acetate further stimulated histone H3 acetylation (+96%) and albumin expression (+473%). In the present study, we further investigate the regulation of these genes by ACSS2 in the presence of acetate and ethanol. Our results provide new insights into how precursors of acetyl‐CoA found in our diet regulate transcription in an ACSS2 dependent manner. The long‐term goal of this study is to identify diet‐dependent epigenetic mechanisms that modulate lipid risk factors for cardiovascular diseases and hepatosteatosis.Support or Funding InformationSupported by UNL Hatch Act, Tobacco Settlement Funds, UNL UCARE Program, and Allen Foundation.
Korean red ginseng extract inhibits p53‐dependent apoptosis in Helicobacter pylori‐infected gastric epithelial cells
Authors: Kang Hyun Ju , Lim Joo Weon , Kim Hyeyoung 
Published: November 2023

Helicobacter pylori (H. pylori) infection is related to the development of gastritis and gastric cancer. H. pylori infection induces oxidative DNA damage, apoptosis and p53 expression in gastric mucosa, which may be involved in gastric carcinogenesis. Prolonged apoptosis may stimulate hyper‐proliferation of neighboring cells.DNA damage responses coordinate DNA repair and apoptosis through the activation of ATM and their target proteins. ATM phosphorylates Ser15 of p53 directly and Ser20 through activation of CHK2, leading to stabilization of p53 and reduces its interaction with MDM2. The level of tumor suppressor protein p53 depends on the levels of MDM2 and ARF. MDM2 is a negative regulator of p53 by its binding to p53. ARF binds and inhibits the activity of MDM2, indicating that ARF disrupts the negative feedback inhibition of p53 by inactivating MDM2. Therefore, prevention of ATM activation, MDM2, and ARF may suppress p53‐mediated apoptosis.Previously, we showed that Korean red ginseng extract (RGE) inhibits NADPH oxidase and reduces the production of reactive oxygen species. H. pylori infection induces oxidative DNA damage and activates DNA damage responses including ATM activation.The present study was purposed to investigate 1) whether H. pylori‐induced apoptosis is mediated with changes in DNA damage response proteins such as ATM, MDM2, ARF, and p53, and 2) whether Korean red ginseng extract (RGE) inhibits H. pylori‐induced apoptosis by regulating DNA damage response proteins in human gastric epithelial AGS cells.The cells were treated with RGE and infected with H. pylori. Cell viability was determined by Trypan blue exclusion assay. Nuclear level of p53 was observed by immunocytochemistry. Protein levels of ARF, MDM2, ATM, p53, Bax, Bcl‐2, and PARP were assessed by Western blot analysis.The results show that H. pylori decreased cell viability, and increased levels of the apoptotic Bax/Bcl‐2 ratio and PARP cleavage. H. pylori increased phosphorylation of ATM and levels of ARF and p53, but decreased MDM2 level. The results suggest that H. pylori‐induced apoptosis is mediated with activation of ATM, decreased MDM2, and increased ARF and p53 in AGS cells. RGE decreased Bax/Bcl‐2 ratio and PARP cleavage, and increased cell viability in H. pylori‐infected cells. RGE inhibited H. pylori‐induced alterations (phosphorylation of ATM, increased ARF and p53, and decreased MDM2) in AGS cells. H. pylori‐induced nuclear translocation of p53 was inhibited by RGE. RGE inhibits p53‐dependent apoptosis by regulating DNA damage response molecules in H. pylori‐infected gastric epithelial cells.In conclusion, Korean red ginseng extract may be beneficial for preventing gastric carcinogenesis associated with H. pylori infection.
Docosahexaenoic Acid Inhibits Cytokine Expression through Suppression of NF‐κB Activation in Rat Pancreatic Stellate Cells
Authors: Eum Hae Rin , Chung Sun Ah , Lim Joo Weon , Kim Hyeyoung 
Published: November 2023

Pancreatic stellate cells (PSCs), when activated by cytokine such as tumor necrosis factor α (TNF‐α), plays an important role in the development of chronic pancreatitis. Induction of inflammatory cytokines and fibrotic proteins contribute to development of chronic pancreatitis. NF‐κB is known to induce pancreatic and systemic inflammatory response. Docosahexaenoic acid (DHA), omega 3‐polyunsaturated fatty acid, shows anti‐inflammatory and anti‐fibrotic effects.In the present study, we investigated whether DHA decreases expression of inflammatory cytokines (MCP‐1, CX3CL1) and fibrotic proteins (alpha smooth muscle actin, α‐SMA; desmin, collagen such as COL1A1 and COL1A1; transforming growth factor beta 1,TGF‐β1) as well as and NF‐κB activity in PSCs exposed to PSCs activating factor TNF‐α.The cells were isolated from pancreas of the rat and cultured in the presence or absence of DHA. mRNA and protein levels were determined by real time PCR and ELISA or western blot analysis.The results show that expression of inflammatory cytokines (MCP‐1, CX3CL1) and fibrotic proteins (α‐SMA, desmin, COL1A1, COL1A1, TGF‐β1), and DNA binding activity of NF‐κB were increased by TNF‐α in PSCs. DHA suppressed TNF‐α‐induced expressions of cytokines and fibrotic proteins by inhibiting NF‐κB activity in PSCs.In conclusion, DHA may inhibit inflammation and fibrosis in pancreatic stellate cells exposed to inflammatory stimulus such as TNF‐α. DHA may be beneficial for preventing development of chronic pancreatitis.Support or Funding InformationThis study was supported by a grant (NRF‐2015R1A2A2A01004855) from the NRF of Korea.