bluuebunny/crossref_metadata_2025_split huggingface dataset:
Separation of α‐retinyl palmitate and quantification of vitamin A after consumption of carrots containing α‐carotene (645.24)
Authors: Goetz Hilary , Kopec Rachel , Riedl Ken , Cooperstone Jessica , Narayanasamy Sureshbabu , Curley Robert , Harrison Earl , Schwartz Steven 
Published: June 2021

Vitamin A (VA) is vital for proper growth, vision, and immune function. Provitamin A carotenoids are an important source of VA for humans, yet the bioequivalence of VA from nonsymmetrical provitamin A carotenoids is poorly characterized. In humans, α‐carotene (αC) is cleaved to VA and inactive α‐retinol (αR) ‐ both of which are esterified with palmitate (P). Analytical methods are needed to separate VA esters from αRP to avoid overestimation of the VA bioequivalence of αC containing foods. To accurately assess the VA obtained from an αC‐rich meal, an αRP standard was synthesized from α‐ionone following adapted methods based on the Wittig‐Horner reaction. A high performance liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) method was developed to resolve αRP from VA esters. Separation was confirmed by MS, where parent ester m/z and differing intensities of daughter ions allowed for distinction of αRP from VA esters. The developed LC‐MS/MS method was applied to the analysis of post‐prandial triglyceride rich lipoprotein fraction of plasma from healthy humans after consumption of carrots and αRP and VA esters were individually quantified. These results provide important insights about the fate of αR in humans consuming αC‐rich foods and the bioequivalence of VA from commonly consumed nonsymmetrical provitamin A carotenoids.Grant Funding Source: Supported by The Ohio State University Food Innovation Center
A role for BCMO1 beyond carotenoid metabolism: regulation of androgen status and signaling (645.4)
Authors: Smith Joshua , Ford NIkki , Clinton Steven , Erdman John 
Published: June 2021

In human studies, higher serum lycopene (Lyc) levels are inversely associated with risk of prostate cancer (PCa), which is driven by androgens. Genomic alterations in carotene‐15,15’‐monooxygenase (BCMO1), which cleaves provitamin A carotenoids to yield retinal, have been shown to alter Lyc accumulation. We hypothesized that Lyc and tomato powder (TP) feeding may impact the prostate by altering testosterone (T) action and that BCMO1 may modulate this process. Nine‐ to fourteen‐week‐old male WT and Bcmo1‐/‐ mice were fed AIN‐93G‐based control or diets containing TP or Lyc for four days. Surprisingly, we found that independent of Lyc or TP feeding, loss of Bcmo1 decreased testicular esterified cholesterol (p<0.05) and testicular mRNA expression of 17β‐hydroxysteroid dehydrogenase 3 (p<0.001), which converts androstenedione to T. In parallel, Bcmo1‐/‐ mice show reduced serum T and reduced androgen receptor‐mediated gene expression in the prostate. Regardless of diet, the weight of the prostate and the seminal vesicles was correlated (R2=0.12, p=0.0015, and R2=0.33, p<0.0001, respectively) with logarithmic concentrations of serum T, in agreement with published androgen‐driven growth kinetics. In summary, BCMO1 seems to play a significant role in androgen and prostate physiology independent of its function in carotenoid metabolism, highlighting a potential role in PCa.Grant Funding Source: Supported by NIH grant PHS‐1‐RO1 CA125384
Flavonoids from rooibos tea promote cell death in an osteosarcoma cell line (647.10)
Authors: Nash Leslie , Ward Wendy 
Published: June 2021

The Saos2 cell culture system is a well characterized osteoblast‐like cell line used to assess the effects of dietary components such as flavonoids on the production of bone mineral. Saos2 cells are derived from human osteosarcoma and possess desirable characteristics for assessing molecular interactions and mechanistic routes in which nutrients may influence mineral production. While screening rooibos tea flavonoids for potential positive effects on bone mineralization, several flavonoids reduced mineralization, while exhibiting cellular death. Mineral, measured using an Alizarin red assay, was reduced in cells treated with isoquercetin (蠅10uM, p<0.001) vitexin (蠅5uM, p<0.05) and luteolin (蠅15uM, p<0.001) in comparison to controls. Metabolic activity of the mitochondrial reductase was analyzed over 72 hours as a marker for cell viability by measuring the reduction of a tetrazolium dye to a formazan salt. The addition of isoquercetin (p<0.001), vitexin (p<0.001) or luteolin (p<0.001) was shown to attenuate mitochondrial activity in a concentration dependent manner, and was in agreement with the mineralization data. Human osteosarcoma is known for its ability to resist chemotherapy. Thus, further investigation of these compounds may help develop natural strategies that enhance chemotherapeutic agents in addition to reducing toxic side effects. Grant Funding Source: Supported by a NSERC Discovery Grant to W. Ward
Synergy of bioactives from camelina seed meal for health promotion (647.16)
Authors: Das Nilanjan , Berhow Mark , Jeffery Elizabeth 
Published: June 2021

Camelina sativa L. Crantz is under development as a novel oil seed crop yet there are few studies about the bioefficacy of camelina. Camelina seed meal contains several potentially bioactive compounds: two major aliphatic GSLs, glucoarabin (9‐(methylsulfinyl)nonylglucosinolate; GSL9), glucocamelinin (10‐(methylsulfinyl)decylglucosinolate; GSL10), with trace amount of a third, 11(methylsulfinyl)undecylglucosinolate (GSL11) and several flavonoids, mostly as quercetin glycosides. We tested hydrolyzed glucosinolates (hGSL) and flavonoids quercetin and rutin for their ability to upregulate detoxification enzymes, as biomarkers of anti‐cancer activity. None of these compounds either caused induction of CYP1A1 activity, or inhibition of β‐naphthoflavone‐induced CYP1A1 activity. In contrast, hGSL9 and hGSL10, quercetin and rutin alone and in combination all caused induction of quinone reductase (NQO1) activity. We found synergistic induction of NQO1 activity when Hepa1c1c7 cells were treated with a combination of all the four major bioactives in the proportions present in camelina seed extract. Although potency of hGSL9 was somewhat less than that of sulforaphane, we found that GSL9 and quercetin appear responsible for this synergy. As Camelina seed extract possesses NQO1 inducing activity, it should be evaluated for anticancer activity, similar to other Brassicaceae family members, such as Broccoli.Grant Funding Source: Supported by USDA/NIFA #AG 2010‐65200
Primary human cardiomyocyte culture: a model for assessment of cardiovascular disorders (652.12)
Authors: Collins Atijah , Harper Emily , Braimah Ayemoba , Sharma Avadhesh 
Published: June 2021

The purpose of this research is to establish an in vitro cardiac system using primary human cardiomyocyte cultures (PHCC). We hypothesized that norepinephrine (NE) and endothelin‐1 (ET‐1) will contribute to human cardiomyocyte growth. PHCC were obtained from Celprogen, San Pedro, CA and grown with varied concentrations of NE or ET‐1 for 72h. Protein expressions of contractile proteins were analyzed via Immunoblot and immunocytochemistry. Analysis of confocal images showed a significant decrease in nuclear area in all NE treatment groups. NE (10 µmol) stimulated significant increases in fluorescent intensities of the contractile proteins, f‐actin, troponin and tropomyosin. The protein expression of myosin light chain was significantly lower with increasing concentrations of NE. In addition, the average cell area increased in all groups of ET‐1. ET‐1 (1, 10, 100 nmol) exhibited a significant decrease of fluorescent intensities of f‐actin, troponin and tropomyosin. PHCC were also grown in the presence of LPS (1, 10, 100µg/ml) in a 6‐well plate for 72h. Viability data showed the cells recovered from the endotoxin treatment and maintain steady growth in each concentration of LPS. In contrast to NE, ET‐1 exhibited a decrease on the expression of contractile proteins in PHCC.Grant Funding Source: Office of Chief Science Officer, PCOM
Role of autophagy related protein 5 induction by globular adiponectin in the suppression of ethanol‐induced apoptosis in liver cells (653.16)
Authors: Nepal Saroj , Kim Mijin , Shrestha Anup , TiliJa pun Nirmala , Park Pil‐Hoon 
Published: June 2021

Hepatocellular apoptosis, an early response of liver cells to hepatic injury on chronic alcohol consumption, has been an area of active research for the treatment of ALD. Adiponectin has been shown to protect liver against chronic ethanol consumption. We have previously demonstrated that ethanol and gAcrp affect expression of genes related with autophagy in liver cells, and further reported the potential utility of autophagy activation by adiponectin in the suppression of ethanol‐induced apoptosis. Herein, we aim to find out whether adiponectin induces autophagy related protein 5 (Atg5), an essential Atg for the elongation of autophagosomal membrane, and if so, explore the role of Atg5 in the suppression of ethanol‐induced apoptosis in liver cells.In the present study, ethanol treatment suppressed Atg5 expression in both primary rat hepatocytes and HepG2 cells while globular adiponectin significantly increased Atg5 expression. Furthermore, pretreatment with globular adiponectin significantly up regulated ethanol‐suppressed Atg5 expression in both primary rat hepatocytes and HepG2 cells.Our initial findings suggest that gAcrp induces Atg5 expression in liver cells that might play a role in the suppression of ethanol‐induced apoptosis.
Experiment‐guided virtual screening provides new high affinity ligands for the benzodiazepine site on GABA‐A receptors (654.4)
Authors: Sigel Erwin , Middendorp Simon , Baur Roland , Puthenkalam Roshan , Ernst Margot 
Published: June 2021

Often novel high affinity ligands are required for a well‐established target protein. Reasons include side effects, resistance development to or sub‐optimal pharmacokinetic or pharmacodynamic properties of an established drug. Here we describe the identification of 19 compounds (14 different scaffolds) that are high affinity ligands for the benzodiazepine‐binding site on mammalian GABA‐A receptors, hitherto un‐crystallized proteins. The procedure includes biochemical techniques that may be generally applied to other proteins. Prerequisites are a ligand that can be chemically modified with cysteine‐reactive groups, knowledge on amino acid residues contributing to the binding pocket on the protein and either crystal structures of proteins homologous to the target protein or even better a crystal structure of the target itself. The procedure includes virtual screening that when used without additional techniques often only results in low affinity ligands, even when a crystal structure of the target protein is available. Combination of crystallization with a functional approach may replace the use of crystallization alone in efforts towards finding new high affinity ligands of an established target.Grant Funding Source: Supported by Swiss National Science Foundation grant 31003A‐132806/1 to E.S.
Synthesis, characterization and toxicity studies of [Ru2(Aap)4cl]: a diruthenium complex (655.12)
Authors: Naidu Naga , Deng Yuanjian , Wei Xin , Yakubu Momoh 
Published: June 2021

Cisplatin has been identified as one of the leading transition metal‐based drug and widely used as anticancer agent. But tumor resistance and development of serious, to overcome these problems we have synthesized a diruthenium complex [Ru2(Aap)4Cl], instrumentally confirm synthesis and evaluate its anti‐proliferative and toxicity effects on A549 cell line and SMC. Aap ligand was synthesized by reaction between 2‐Chloro‐4‐aminopyridine and excess amount of aniline. The Aap was reacted with Ru2(CH3COO)4Cl to yield the Ru2(Aap)4Cl complex. The ligand and the diruthenium complex were analyzed for their thermal, elemental, spectroscopic (NMR (1H & 13C NMR), IR, and MS) and structural properties. Cell proliferation, viability and toxicity were performed using the MTT assay. Spectroscopic and thermal data were consistent with the proposed structures, single X‐ray crystallography of the ligand and the complex further revealed that: I) the 4‐amino group on the pyridine ring was not involved in complex formation; ii) the diruthenium complex was a (4, 0) isomer; and iii) interesting conformational changes were also observed in the Aap ligand after the complex formation. The anti‐proliferative property of the new diruthenium complex was evaluated at 24, 48, and 72 hrs following treatment and showed that the [Ru2(Aap)4Cl] complex reduced cell proliferation by 50% with an IC50 value of 5 ±1 μM for A549 cell line, and 75 ± 5 μM for SMC. These results suggests that the Ru2(Aap)4Cl complex is a promising anticancer agent and needs further studies to fully evaluate the anticancer properties by evaluating the molecular mechanism by which it reduces cellular proliferation.Grant Funding Source: supported by grants from NHLBI: HL03674 and HL070669.
Epigallocatechin‐3‐gallate sensitizes therapeutic efficacy of doxorubicin involving autophagy inhibition in hepatoma Hep3B cells (655.2)
Authors: Ye Hui‐Lan , Chen Li , Zhang Guo , Chen Xing‐Zhou , Yao Wen‐Min , Zhang Fa‐Can 
Published: June 2021

The green tea component (‐)‐Epigallocatechin‐3‐O‐gallate(EGCG), has promisingly been found to enhance the efficacy of several chemotherapy agents in hepatocellular carcinoma (HCC) treatment and its detailed mechanisms remain unclear. Herein, this study aimed to confirm the synergistic interaction of EGCG and DOX on Hep3B cells and to define whether the autophagic flux is involved in this cooperated fashion. Firstly, by MTT and trypan blue assay, both EGCG and DOX exhibited cellular growth inhibition and cell death induction effects. However, by western blotting and real time RT‐PCR, DOX dramatically induced the expression of Atg5 and beclin1, the autophagic signaling molecules, whereas EGCG presented a markedly and dose‐dependently inhibitory role in autophagy signaling. Secondly, EGCG and DOX combined treatment amazingly caused a synergistic effect of about 50% increment on cell death and about 45% augmentation on apoptosis versus monotherapy pattern. Interestingly, the DOX‐induced autophagy was abolished by this combination therapy. Thirdly, the anticancer effect of either DOX or combination with EGCG treatment was substantially impaired by either rapamycin, an autophagic agonist, or Atg5 as well as beclin1 siRNAs. Consistently, significantly tumor growth reduction and apoptosis increment were found in combination therapy compared with DOX alone in vitro study. And immunohistochemisty analysis indicated that the suppressed tendency of autophagic hallmark LC3 expression was consistent with thus combined usage in vitro. Taken together, the current study suggested that EGCG emerges as a chemotherapeutic augmenter and synergistically enhances DOX anticancer effects involving autophagy inhibition in HCC.Grant Funding Source: Supported by NSFC Grants 881172260, 30630145, 81360077 to G.Z. & 81102495 to L.C.
Nrf2‐Keap1 pathway does not have an important role in cisplatin resistance in neuroblastoma (655.8)
Authors: Tanoshima Reo , Yang MingDong , Dalvi Pooja , Wu Alex , Matsuda Kensuke , Zhang Libo , Fujii Hisaki , Baruchel Sylvain , Harper Patricia , Ito Shinya 
Published: June 2021

Drug resistance is one of the major causes of treatment failure in neuroblastoma (NB). Cisplatin is one of the key chemotherapeutic agents for NB. Nuclear factor erythroid 2‐like‐2 (Nrf2) is a transcription factor that regulates anti‐oxidative stress enzymes and drug transporters, negatively regulated by Keap1 (kelch‐like ECH‐associated protein 1). Loss‐of‐function mutations of Keap1 gene and subsequent Nrf2 activation have been shown to be a mechanism of cisplatin resistance in adult cancers. We conducted this study to clarify the role of Nrf2‐Keap1 pathway in cisplatin resistance in NB. We investigated the basal activity of Nrf2 in nine NB cell lines (IMR‐32, LAN5, CHLA‐15, SH‐SY5Y, CHLA‐20, SKN‐FI, SK‐N‐BE(2), BE(2)C, CHLA‐90) using Antioxidant Response Element luciferase reporter assay. No cell lines had high reporter activities. In order to evaluate whether cisplatin activates Nrf2, we analyzed the mRNA expression of Nrf2 and its target genes (Glutamate‐Cysteine Ligase, Modifier Subunit: GCLM and NAD(P)H Dehydrogenase, Quinone 1: NQO1) after 24h treatment with cisplatin. In this experiment, we identified CHLA‐90 as one of the 3 cell lines, which showed high induction of Nrf2 target genes upon cisplatin exposure. However, knockdown of Nrf2 expression by small interfering RNA did not improve MTT assay‐measured dose‐response survival rate of CHLA‐90 with cisplatin treatment.We conclude that the Nrf2‐Keap1 pathway does not necessarily play an important role in cisplatin resistance in NB cells.Grant Funding Source: Supporter by CIHR, TCCSG, and Joseph M. West Family Memorial Fund
Effects of a heroin vaccine on heroin self‐administration and its pharmacokinetic correlates (658.5)
Authors: Raleigh Michael , Pentel Paul , LeSage Mark 
Published: June 2021

Vaccination with a heroin vaccine (M‐KLH) produced a compensatory increase in heroin self‐administration (HSA) at high heroin unit doses, but a decrease in HSA at low unit doses. Vaccination also blocked heroin‐primed reinstatement of HSA responding during extinction. To study the PK correlate of vaccine‐induced increases in HSA at high unit doses, repeated i.v. heroin injections were administered to vaccinated and control rats that simulated heroin exposure during HSA (0.5 mg/kg/hr heroin in control rats and 1.0 mg/kg/hr heroin in vaccinated rats). 6‐monoacetylmorphine (6‐MAM) and morphine, but not heroin, distribution to brain were reduced in vaccinated rats compared to controls. 6‐MAM concentrations in brain were similar between control rats given 0.5 mg/kg and vaccinated rats given 1.0 mg/kg, suggesting that 6‐MAM was the main heroin metabolite mediating compensation of HSA. These data show that vaccination with M‐KLH altered HSA and reinstatement, that 6‐MAM concentrations in brain mediate these effects, and that high antibody to drug ratios are most effective at reducing heroin’s reinforcing effects.Grant Funding Source: Supported by DA026300, DA030715, and DA07097
Characterization of the kappa opioid receptor‐mediated G protein signaling in mouse striatum using a [35S]GTPγS binding assay (659.6)
Authors: Zhou Lei , Stahl Edward , Lovell Kimberly , Slauson Stephen , Frankowski Kevin , Aube Jeffrey , Bohn Laura 
Published: June 2021

The kappa opioid receptor/dynorphin system is an endogenous system involve in modulating stress responses. Disruption of the ability to respond appropriately to stress can lead to elevated risks of developing mood disorders including depression, anxiety, and drug addiction. The dynorphin peptides are products cleaved from prodynorphin of varying amino acid length and are considered the cognate high affinity ligands for the KORs. However, they also promiscuously bind to and activate other opioid receptors in the brain. Here we seek to determine what component of dynorphin signaling can be attributed specifically to KOR by investigating the pharmacological profiles of dynorphin peptides in wild‐type (WT), KOR knockout (KOR‐KO), and mu opioid receptor knockout (MOR‐KO) mice using [35S]GTPγS binding assay in striatal membrane preparations. We find that dynorphin peptides are efficacious in stimulating [35S]GTPγS binding in WT striatum in a manner that resists concentration‐induced plateau. While KOR activation contributes significantly to dynorphin‐induced [35S]GTPγS binding in striatum, our studies identify a significant contribution of MOR‐induced coupling as well. Moreover, we find that KOR displays a constitutive activity in striatum that can be inhibited by antagonist pretreatment; the constitutive activity is not present in KOR‐KO striatum. Recently developed, functionally selective small molecule KOR agonists as well as new selective KOR antagonists were also tested in striata. Our studies in characterization of dynorphin’s effects at the KOR in brain will be essential for developing ex vivo assays that assess the selective contributions of biased KOR ligands that have been recently developed.Grant Funding Source: Supported by NIDA grant R01 DA031927(LMB, JA)
Tolerance and cross‐tolerance to behavioral effects of phenethylamine‐ and tryptamine‐derived hallucinogens in mice (661.2)
Authors: Smith Douglas , Williams Diarra , Fantegrossi William 
Published: June 2021

Tolerance refers to a decrease in effect(s) of a drug due to repeated administration, and develops rapidly to psychoactive effects of some hallucinogens in man. Tolerance to various effects of hallucinogenic drugs can also be readily induced in laboratory rodents, and is presumably mediated by downregulation of serotonin 5‐HT2A receptors. The hallucinogen‐induced head twitch response (HTR) in mice is a reliable rodent model for psychedelic effects because it is mediated by agonist activity at 5‐HT2A receptors, but the role of tolerance in HTR remains somewhat controversial. In these studies, two phenethylamine hallucinogens (2,5‐dimethoxy‐4‐iodoamphetamine [DOI] and 2,5‐dimethoxy‐4‐n‐propylthiophenethylamine [2C‐T‐7]) and two tryptamine hallucinogens (dipropyltryptamine [DPT] and diisopropyltryptamine [DIPT]) were administered to male NIH Swiss mice to determine dose‐effect functions for HTR. The most effective dose of each compound was then selected for repeated administration to assess the development of tolerance to the HTR. Finally, mice received daily treatment with DOI for 3 days, then were substituted with different doses of DOI, 2C‐T‐7, DPT or DIPT the next day to assess surmountability of tolerance and cross‐tolerance among drugs. Repeated administration of the phenethylamines ‐ but not the tryptamines ‐ resulted in rapid tolerance to HTR in mice which was not surmountable with increased dose. Although the tryptamines did not induce tolerance to HTR with repeated administration, cross‐tolerance to HTR elicited by DPT was observed in mice previously treated with DOI, and this cross‐tolerance was not surmountable with increased DPT dose. Explanations for these distinct effects of phenethyltamine‐ and tryptamine‐based hallucinogens may include the involvement of 5‐HT2C receptors (for the phenethylamines) or 5‐HT1A receptors (for the tryptamines.)Grant Funding Source: Supported by RR020146, RR029884, and by an ASPET SURF award to DS