bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cranberry extract initiates apoptosis in HL‐60 cell line through intrinsic apoptotic pathway (644.9)
Authors: Mansouri Rasha , Khoo Christina , Percival Susan 
Published: June 2021

The potential anticancer activity of cranberry was reported for the first time in 1996. Proanthocyandins (PACs), the most abundant and complex flavonoid in cranberry, have been described as the major contributors of the anticancer activity reported in the literature. One of the proposed chemopreventative mechanisms is the induction of apoptosis by cranberry PACs. However, limited studies have been done to investigate the role of cranberry in initiating apoptosis in cancer cells and its underlying mechanism of action remains undetermined. Therefore, the overall aim of this study was to investigate the apoptotic pathway that cranberry affects, intrinsic vs. extrinsic pathway. In our study, HL‐60 cells were incubated with 25 µg/ml cranberry‐(60‐65%)PAC extract for 24 hours. Cranberry treatment showed an 85 % increase in caspase 3/7 activity. The data also showed a great enhancement in the activation of caspase‐9 (379 %), but it did not exhibit any change in caspase‐8 activity. Further tests revealed that cranberry treatment caused a loss of mitochondrial transmembrane potential leading to an increase in the mitochondrial release of cytochrome C (87.5%) and Smac (41%). These findings indicate that this cranberry‐PAC extract can induce apoptosis in HL‐60 cells through triggering the intrinsic (mitochondrial) apoptotic pathway.Grant Funding Source: Funded by Agricultural Experiment Station and Ocean Spray Cranberries, Inc.
Consuming iron‐biofortified pearl millet increased hemoglobin concentrations and prevented a decline in energy efficiency in Indian girls (646.7)
Authors: Beer Hannah , Luna Sarah , Pompano Laura , Przybyszewski Eric , Udipi Shobba , Ghugre Padmini , Haas Jere 
Published: June 2021

Objective: To determine if the change in iron status from an iron biofortified efficacy trial results in a change in energy efficiency for Indian girls.Methods: 56 Indian girls (12‐15 years) were randomly assigned to receive iron biofortified pearl millet or traditional pearl millet for 6 months. Blood and energy efficiency measurements were taken before and after the intervention. Energy efficiency was calculated by recording O2 and CO2 consumption during sub‐maximal exercise tests on a Monark 874E ergometer. General linear models were used to evaluate the effect of the iron biofortification intervention on energy efficiency of the girls.Results: Consuming iron biofortified pearl millet resulted in an increase in hemoglobin after 3 months (p=0.05). Energy efficiency decreased in both treatment and control groups, but was significantly higher in the treatment group (p=0.02).Conclusion: Consuming biofortified pearl millet improved hemoglobin status in Indian girls and prevented a decline in energy efficiency over 6 months.Grant Funding Source: Supported by HarvestPlus
Techological valorization of anthocyanin‐rich Brazilian berries (647.36)
Authors: Alves Andressa , Lacerda Ellen , Inada Kim , Silva Laís , Silva Tamirys , Castelo‐Branco Vanessa , Finotelli Priscilla , Torres Alexandre , Monteiro Mariana , Perrone Daniel 
Published: June 2021

This study aimed to add value to two anthocyanin‐rich Brazilian berries by technological processes. Jabuticaba (Myrciaria cauliflora) and Jussara (Euterpe edulis) juices were pressurized at 200 MPa for 5 and 10 min. Jussara juice was microencapsulated by spray drying using Capsul® and Capsul® and inulin (1:1) as encapsulating agents. Jussara oils were obtained by cold extraction with either petroleum ether (JPO) or ethanol (JEO). Anthocyanin contents in products were determined by HPLC‐UV and tocopherols contents in oils were determined by HPLC with fluorescence detection. Jussara products contained both cyanidin‐3‐O‐glucoside and cyanidin‐3‐O‐rutinoside, whereas Jabuticaba juices contained only the former anthocyanin. Although pressurization of Jussara juice for 5 min led to a decrease of 19% in anthocyanins in comparison to control (3.87 g/L), the content was restored when the juice was pressurized for 10 min. In contrast, pressurization of Jabuticaba juice for 5 min led to an increase of 29% in anthocyanins in comparison to control (1.11 mg/L), but this increase was not maintained after pressurization for 10 min. Capsul®:inulin microcapsules showed higher contents of both cyanidin‐3‐O‐glucoside (7.14 g/kg) and cyanidin‐3‐O‐rutinoside (8.98 g/kg) than those prepared with Capsul® alone (5.00 and 5.73 g/kg, respectively). Although JPO extraction yield was higher (48.9%) than that of JEO (37.7%), the latter showed a 24‐fold higher anthocyanins content (21.1 and 22.1 mg/100 g of cyanidin‐3‐O‐glucoside and cyanidin‐3‐O‐rutinoside, respectively). Tocopherols contents in JEO were 1.6‐fold higher (150.9mg/100g) than in JPO, with alpha‐ and delta‐tocopherols corresponding to 56% and 40% of total tocols, respectively. As a whole, our results suggest that pressurization, microencapsulation and oil extraction are suitable technologies for Brazilian berries’ valorization.Grant Funding Source: FAPERJ, CNPq, CAPES, UFRJ
Activation of mitochondrial‐u‐calpain sensitizes opening of the mitochondrial permeability transition pore during ischemia‐reperfusion (648.11)
Authors: Chen Qun , Hu Ying , Lesnefsky Edward 
Published: June 2021

BACKGROUND: Opening of the mitochondrial permeability transition pore (MPTP) increases cardiac injury during ischemia‐reperfusion (IR). IR activates both cytosolic and mitochondrial calpains. Administration of MDL‐28170 (MDL, a calpain inhibitor) decreases injury in isolated hearts during IR. We asked if inhibition of calpains during IR decreases MPTP opening and if activation of mitochondrial u‐calpain (m‐u‐calpain) predisposes to MPTP.METHODS: Buffer‐perfused mouse hearts (C57BL/6) were subjected to 25 min ischemia and 30 min reperfusion with or without MDL (10 uM) treatment. Opening of MPTP was evaluated by calcium retention capacity (CRC, nmol calcium/mg protein) in mitochondria isolated from IR hearts. In control mitochondria (protease‐purified to remove cytosolic contamination), exogenous calcium was used to activate mito‐u‐calpain. The ADP‐stimulated change in inner membrane potential (ΔΨ) during the transition from state 4 to state 3 respiration (reflected by alteration in TMRM fluorescence arbitrary units, A.U.) was used to assess inner membrane integrity.RESULTS: Inhibition of calpains improved CRC in mitochondria following IR, supporting that calpain activation enhanced MPTP opening. In control mitochondria, exogenous calcium progressively depolarized ΔΨ, whereas inhibition of m‐u‐calpain attenuated the extent of depolarization.CONCLUSION: Inhibition of calpain in purified mitochondria reduces the calcium‐induced permeation of the inner membrane, indicating that activation of m‐u‐calpain likely contributes to MPTP opening and cardiac injury during IR.

Isolated hearts
LDH release
LVDP
CRC

IR
615±47
13±2
424±50

MDL + IR
383±55*
29±5*
560±16*

Isolated mito
Î”Î¨, TMRM, A.U.

Calcium uM
0
50
100

DMSO + mito
0.088±0.009
0.050±0.003â€
0.013±0.003â€

MDL+ mito
0.087±0.006
0.066±0.004â€¡
0.036±0.013â€¡

Mean ± SEM; LVDP‐developed pressure mmHg; LDH‐mU/g/min; *p<0.05 vs. IR; †p<0.05 vs. calcium (0 uM); ‡ p<0.05 vs. untreated mitochondria (mito).Grant Funding Source: Supported by Scientist Development Grant from AHA
Activation of HIF‐1 pathway by cholesterol in hepatocytes under normoxia (649.1)
Authors: Anavi Sarit , Hahn Obercyger Michal , Madar Zecharia , Tirosh Oren 
Published: June 2021

Hypoxia inducible factor‐1 (HIF‐1) and cholesterol have been separately implicated in the development of several liver pathologies. The aim of the present study was to investigate possible activation of HIF‐1 pathway by cholesterol in AML12‐hepatocytes.Increased HIF‐1α protein accumulation was evident in hepatocytes after 2h of incubation with cholesterol while enhanced HIF‐1 transcriptional activity was observed after 6h. Investigating the molecular mechanism revealed that cholesterol inhibited HIF‐1α proteasomal degradation. Hepatocytes incubated with cholesterol exhibited enhanced Nitric oxide (NO) levels, which corresponded with increased iNOS induction. Inhibition of iNOS had no effect on HIF‐1α stabilization at 2h but effectively attenuated cholesterol‐induced HIF‐1 accumulation and transcriptional activity at 6h. Mitochondrial dysfunction and enhanced mitochondrial ROS generation were observed in 2h cholesterol‐treated cells. In order to establish a role for mitochondria‐derived ROS in HIF‐1α protein stabilization, mitochondrial DNA‐depleted cells were utilized. In these cells, the ability of cholesterol to induce HIF‐1 pathway was significantly abolished.These results demonstrate that cholesterol promotes HIF‐1 activation in hepatocytes in a ROS‐dependant manner. Therefore, alterations of HIF‐1 may contribute to the deleterious effects of cholesterol in the liver.
Garcinol attenuates TGF‐β1/Smad signaling in dimethylnitrosamine‐induced liver fibrosis (649.8)
Authors: Cheng An‐Chin , Lee Ming‐Fen , Tsai Chen‐Yu , Li Chun‐I , Pan Min‐Hsiung 
Published: June 2021

Garcinol is a polyisoprenylated benzophenone derivative from Garcinia indica and other related species. Many studies have shown that garcinol is a pleiotropic agent against inflammation. Our previous studies indicated that Garcinol supplementation alleviated the dimethylnitrosamine (DMN)‐induced liver damage, including the expression levels of alanine transaminase and aspartate transaminase and the histopathological status. In this study, we further investigate the effect of garcinol on the transforming growth factor‐β1 (TGF‐β1)/Smad signaling. Sprague‐Dawley rats were intraperitoneally given DMN (10 mg/kg) three days per week for four weeks. Garcinol (10 or 20 mg/kg) was administered by oral gavage daily. Hepatic tissue sections were stained for α‐smooth muscle actin (α‐SMA) and collagen I, and the hepatic TGF‐β1 expression was examined. Our results indicated that garcinol suppressed collagen deposition and α‐SMA formation in the DMN‐treated animals. These data demonstrated that garcinol attenuated TGF‐β1/Smad signaling in liver fibrosis.Grant Funding Source: Supported by Chang Jung Christian University Grant
Serine protease modulation of epithelial barrier function through the tight junction (650.8)
Authors: Ronaghan Natalie , MacNaughton Wallace 
Published: June 2021

Inflammatory mediator‐induced disruption of the tight junction may contribute to pathogenesis of IBD through increased intestinal permeability. We have previously found apical addition of the serine proteases trypsin and matriptase to colonic epithelial cell lines increase transepithelial electrical resistance (TER). We aimed to elucidate the role of tight junction (TJ) modulation in this phenomenon. Confluent monolayers of SCBN colonic epithelial cells were mounted on Ussing chambers after being transfected with siRNA against occludin and treated with 45 BAU/mL trypsin. Trypsin‐induced changes in TER and permeability to 4 kDa FITC‐dextran were monitored. siRNA reduced occludin levels by 90% (Western blot) and reduced the trypsin‐induced increase in TER to 29.8 ± 6.0% (n=4 p<0.05) of control response. However, no changes in trafficking of any TJ protein were observed by Triton X‐100 fractionation and Western blotting after trypsin treatment. In a calcium‐switch assay performed in Ussing chambers, trypsin did not induce faster recovery or have an affect on TER until 30 minutes post‐recovery of the monolayer. In conclusion, we have shown that occludin is essential in the serine protease‐mediated increase in TER but this is not through trafficking to the TJ, and that trypsin does not induce the formation of the tight junction.Grant Funding Source: Supported by Canadian Institutes of Health Research
Oxidative stress induces apoptosis of MCF7 cells treated by HDAC inhibitors (655.13)
Authors: Hirata Aiko , Senanayake Thulani , Woster Patrick , Hirata Fusao 
Published: June 2021

When MCF7 human breast adenocarcinoma cells were treated by HDAC inhibitors (HDACi), SAHA or THS79‐12, considerable portions of MCF7 cells became floated and underwent apoptosis. While annexin A1 was induced in both floated and attached cells, this protein in floated cells was mono‐ubiquitinated as determined by Western blots, suggesting that DNA damage took place. PML, a protein that plays an important role in DNA damage induced apoptosis was acetylated, and Bclxs was induced, and caspase 3 was cleaved, demonstrating features of apoptosis.8‐Oxoguanosine contents of cells treated with HDAC inhibitors increased by 8 folds in floating cells as compared with those in attached cells. These apoptotic changes did not occur in attached cells treated with HDACi, but their growth was suppressed as seen with annexin A1 transfected MCF7 cells. These observations suggest that apoptosis of MCF7 cells treated with HDACi is attributed to oxidative stress induced by HDACi rather than to annexin A1 induction by HDACi.Grant Funding Source: Supported in parts by a grant from The Susan G. Komen Breast Cancer Foundation.
A novel nociceptin receptor antagonist inhibits feeding behavior in rodents (656.1)
Authors: Statnick Michael , Toledo Miguel , Witkin Jeffrey , Song Min , Hu Chalie , Ansonoff Michael , Pintar John , Lafuente Celia , Alma Jiménez Alma , Ana Benito Ana , Martínez‐Grau Mariangles , Pedregal Conception , Chen Yanyun 
Published: June 2021

Background: Nociceptin is a 17 amino acid peptide that produces physiological effects through activation of the ORL1 (NOP) receptor. We have identified a novel NOP antagonist (Cpd 1) and found it to have high binding affinity (Ki = 0.11 nM), antagonist potency (Kb = 0.17 nM) and selectivity for NOP. Cpd 1 is orally active, CNS penetrant and potently occupies NOP receptors in vivo. Herein, the effects of Cpd 1 on feeding behavior and metabolism were studied.Methods: Male, lean and dietary‐induced obese (DIO) rats, DIO mice and NOP knockout (KO) mice were used in feeding and indirect calorimetry studies.Results: Consistent with the literature, Cpd 1 reduced food intake in rodents. In lean rats, Cpd 1 inhibited consumption of a highly palatable (high fat/sucrose) diet, reducing the calorie intake to control chow levels. In DIO rats, Cpd 1 inhibited feeding induced by a mild caloric restriction. In DIO mice, Cpd 1 decreased 24 hr free access food intake and decreased the respiratory quotient indicating that metabolism was driven by fat utilization. Fasting‐induced feeding was inhibited by Cpd 1 in 129S6 mice, an effect that was absent in NOP KO mice. Moreover, under vehicle conditions fasting‐induced feeding was reduced in NOP KO mice.Conclusions: These data support that Cpd 1 potently inhibits feeding behavior and stimulates fat utilization in rodents. Moreover, the hypophagic effect of Cpd 1 is dependent on NOP receptors.Grant Funding Source: Eli Lilly and Company
Critical interaction of the dopamine D1 receptor with ergolines: the roles of conserved aromatic residues in transmembrane six segment (656.5)
Authors: Lee Sang‐Min , Mailman Richard 
Published: June 2021

We have investigated the interaction of the dopamine D1 receptor with ergoline‐based ligands to understand how non‐catechols activate this receptor. Here, we examined the roles of conserved aromatic residues (F6.51 and F6.52) in transmembrane six segment that are reported to be conserved sites for receptor activation. Mutations of F6.51 and F6.52 to alanine (F6.51A and F6.52A) dramatically decreased the binding affinity of ergolines. Consistent with the affinity data, the functional potency of these ligands at D1‐mediated cAMP synthesis was eliminated at the F6.51A receptor. Whereas F6.52A significantly reduced the potency of these ergolines (consistent with their decreased affinity), it did not decrease the maximal cAMP production. We also studied F6.51Y and F6.51W that were more likely to conserve aromatic/hydrophobic interactions of F6.51. Although F6.51Y did not restore the affinity decrease by F6.51A, it partially restored the potency and fully restored cAMP production. F6.51W was essentially identical to the wild‐type receptor in terms of both affinity and activity of the ergolines. Using a D1 receptor homology model, we hypothesize that the B and D ring of the ergoline backbone has a docking pose close to F6.52 and F6.51, respectively. The F6.51W mutation mimics the hydrophobic interaction that ergolines have with the wild‐type receptor. These data can be useful for designing non‐catechol agonists for the D1 receptor. Grant Funding Source: Supported by Grant PDF‐SFW‐1208 from the Parkinson's Disease Foundation
Probing partial/biased agonism at the G protein‐coupled A2B adenosine receptor (662.11)
Authors: Gao Zhan‐Guo , Balasubramanian Ramachandran , Kiselev Evgeny , Jacobson Kenneth 
Published: June 2021

The A2B adenosine receptor (AR) is involved in many important functions and couples to several G proteins, but the endogenous effects have not been well documented partly due to the lack of selective agonists at this subtype until recently. In the present study, we probed the potential partial and/or biased agonism at the A2B AR in several types of cells, including HEK293 cells and T24 bladder cancer cells, using a series of A2B agonists of adenosine analogs substituted at 5’, N6 and C2 positions, including NECA, MRS5911 and MRS3997, and the non‐nucleoside A2B agonist, BAY60‐6583. All A2B agonists tested induced accumulation of cyclic AMP fully or partially in all cells tested, with NECA being the most efficacious. However, only 5’‐substituted analogs are full agonists in calcium mobilization and ERK1/2 phosphorylation. Over‐expression of the A2B AR in HEK293 cells increased the agonist potency and efficacy in inducing the Gs‐mediated accumulation of cyclic AMP, but not the ability to induce calcium mobilization by the C2‐substituted adenosine derivatives or non‐adenosine A2B agonists. Results from an arrestin translocation assay showed that the 5’‐substituted NECA is more efficacious than adenosine derivatives substituted at C2 and N6 positions and non‐adenosine agonists. BAY actually did not induce arrestin translocation in this assay. The data suggest that 5’‐substituted adenosine analogs are balanced agonists of different pathways, whereas the C2‐ and N6‐substituted adenosine analogs and the class of non‐nucleoside A2B agonists are Gs‐biased agonists. The data also suggest that BAY and some N6‐ and C2‐substituted adenosine analogs may be full agonist, partial agonist or antagonist under various conditions. Indeed, BAY behaves as an A2B antagonist to induce insulin release in MIN6 pancreatic β cells. Thus, differences in efficacy and effector pathways might explain some of the contradictory results reported for A2B AR activation.Grant Funding Source: Supported by NIDDK Intramural Research, NIH
Identification of a novel, highly potent D3 dopamine receptor‐selective agonist (662.8)
Authors: Moritz Amy , Free R. , Conroy Jennie , Weiner Warren , Doyle Trevor , Southall Noel , Ferrer Marc , Javitch Jonathan , Aubé Jeffrey , Frankowski Kevin , Sibley David 
Published: June 2021

Dopamine receptors (DARs) are involved in the development and/or treatment of many neuropsychiatric disorders including schizophrenia and Parkinson’s disease. Many currently available dopaminergic drugs modulate both D2 and D3 DARs due to high homology in their orthosteric binding sites, leading not only to potential unwanted side effects, but also uncertainty as to the roles each DAR subtype plays in normal and pathological processes. In order to discover compounds that target unique, less conserved allosteric sites of these DARs, our lab employed a high throughput screening approach. Through the NIH Molecular Libraries Program, compound 3843 was originally identified as a D2 antagonist in a screen of a 380,000+ small molecule library. Counter‐screening assays of beta‐arrestin recruitment revealed that this compound selectively activates the D3 DAR, yet also acts as an antagonist at the D2 DAR. Over 270 analogs were synthesized and tested to explore the structure‐activity relationship of 3843 for the D2 and D3 DARs, and several compounds with a 1,000‐fold or greater increase in D3 DAR agonist potency were found. Initial competition binding assays suggest these compounds act in an allosteric manner at the D3 DAR, and initial ADME studies performed on 3843 were favorable. We ultimately hope these probes will be useful in vitro and in vivo pharmacological tools to elucidate D3 DAR specific physiology and pathology.Grant Funding Source: Supported by NIH IRP
Mitochondrial telomerase regulates flow mediated dilation by suppressing mitochondrial derived free radical production (664.1)
Authors: Santos Janine , Gutterman David , Beyer Andreas 
Published: June 2021

Mitochondrial (mt) reactive oxygen species (ROS) are key modulators of vascular reactivity. Coronary artery disease (CAD) changes the mechanism of flow‐mediated dilation (FMD) from NO to mtH2O2. Decreased telomerase activity, prominent in cellular senescence and tissue aging, correlates with mtROS, and pharmacological inhibition of telomerase mirrors this change. We hypothesize that telomerase regulates the mechanism of FMD by mitigating mtROS.FMD, analyzed by video microscopy, was reduced in mesenteric arteries (~200 µm) of mice lacking the catalytic subunit of telomerase (TERT‐/‐). (% Max dilation at 100 cmH2O WT 79.8 ±5.2; TERT‐/‐ 42.5 ±9.6; n蠅6 P< 0.05 ANOVA). L‐NAME inhibited FMD in controls whereas in TERT‐/‐ catalase blocked dilation indicating NO and H2O2 as mediators respectively.
TERT is present both in nucleus and mitochondria. To differentiate the mtROS‐decreasing activity as requiring the nuclear vs. mt activity of TERT, we used cultured human fibroblast expressing WT or a mutant TERT unable to localize to mitochondria (nucTERT). nucTERT cells had higher mtROS, decreased MnSOD protein levels and produced less ATP, which paralleled mitochondrial ultrastructure abnormalities. These functional changes were also observed in endothelial cells mitochondria from TERT‐/‐. Taken together, these data indicate that mitochondrial TERT is a key regulator of mtROS in the microvasular endothelium.
Transformed human brain pericytes enhance endothelial wound healing and angiogenic sprout formation (670.5)
Authors: Mayo Jamie , Bearden Shawn 
Published: June 2021

We have recently discovered that density‐stressed human brain pericytes transform to a stem cell‐like phenotype; they form spheres that self‐renew and generate daughter cells with varying morphologies. Co‐culture of transformed pericytes with endothelial cells in a scrape wound assay increased the rate of wound healing as compared with non‐transformed co‐cultures. The goal of this study was to identify mechanisms by which transformed pericytes increase the rate of wound healing and to test the hypothesis that transformed pericytes increase the rate of angiogenic sprout formation. To stimulate sprout formation, primary human brain endothelial cells and pericytes were co‐cultured in a collagen matrix with vascular endothelial growth factor. In a scrape wound assay, lipophilic cell membrane dyes (DiO, DiD; Molecular Probes) were used to visualize and track respective cell types. Transformed pericytes increased sprout formation by 2.13 ± 0.08 fold (p<0.05) compared to endothelial cells co‐cultured with non‐transformed pericytes. In the scrape wound assay, both endothelial cells and transformed pericytes were present at the wound edge. In contrast to the non‐transformed co‐cultures, many of the transformed pericytes fused with endothelial cells during wound repair. Cell fusion was positively correlated with a higher rate of wound healing. Transformed pericytes may be a source of stem cell‐like cells with pro‐angiogenic potential.Grant Funding Source: NIH HL10654801