bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison of mineral content in breast milk between early and later lactation periods among indigenous women from Western Highlands of Guatemala (623.11)
Authors: Li Chen , Wren Hilary , Xu Ran , Solomons Noel , Chomat Anne Marie , Scott Marilyn , Koski Kristine 
Published: June 2021

Background: Human breast milk is the normal source of minerals for infant growth, development and metabolic function up to 6 months, but little information is available about how mineral concentrations vary throughout lactation in developing countries. Our objectives were to compare mineral concentrations between early (<45d) and later (4‐6mo) lactation periods, and to determine if mineral concentrations varied with infant age within these two lactation periods.Methods: Inductively Coupled Plasma Mass Spectrometry was used to analyze the concentration of 13 minerals (Na, K, Ca, Mg, Mn, Zn, Cu, Cr, Sr, Se, Rb, Fe, P) in early (n=54) and later (n=49) breast milk samples collected from Mam‐Mayan women from rural villages in the Western Highlands of Guatemala.Results: Median concentrations of Na, K, Mn, Zn, Cu, Cr, Rb and Fe were significantly higher in early milk compared with later milk whereas Mg was lower in early milk. No differences were observed between lactation period for Ca, Se and Sr. Within the first 45d, Na, K, Cu and Zn were negatively correlated with infant age. During the 4‐6mo interval, only Se was negatively correlated with infant age.Conclusion: Our findings reveal that concentrations of Na, K, Cu and Zn decreased within the first 45d, that Mn, Cr, Rb and Fe had declined by 4‐6mo, and that Se declined during the 4‐6mo interval.Grant Funding Source: McGill University International Mobility Award, NSERC
The synergistic effect of 1’‐acetoxychavicol acetate and sodium butyrate on the death of human hepatocellular carcinoma cells (644.17)
Authors: Matsui‐Yuasa Isao , Kato Rie , Azuma Hideki , Kojima‐Yuasa Akiko 
Published: June 2021

It has been suggested that the combined effect of natural products may improve the effect of treatment against the proliferation of cancer cells. We evaluated the combination of 1’‐acetoxychavicol acetate (ACA), obtained from Alpinia galangal, and sodium butyrate (NaB), a major short chain fatty acid, on the growth of HepG2 cells and found that treatment had a synergistic inhibitory effect.The number of HepG2 cells was synergistically decreased via apoptosis induction with the combined treatment of ACA and NaB. In ACA‐ and NaB‐treated cells, intracellular reactive oxygen species (ROS) levels and NADPH oxidase activities were increased significantly. The decrease in cell number after combined treatment of ACA and NaB was improved with the treatment of catalase. These results suggest that an increase in intracellular ROS levels is involved in cancer cell death. AMP‐activated protein kinase (AMPK) plays an essential role in controlling processes related to tumor development. In ACA‐ and NaB‐treated cells, AMPK phosphorylation was induced significantly, and this induction improved when cells were pretreated with catalase. These results suggest that the increase in intracellular ROS is involved in the increase of AMPK phosphorylation.In conclusion, combined treatment with ACA and NaB synergistically induced apoptotic cell death via an increase in intracellular ROS and phosphorylation of AMPK.
Peptides in common bean (Phaseolus vulgaris L.) non‐digestible fraction inhibit human colorectal cancer cell survival in vitro through oxidative stress injury: a comparative study (644.5)
Authors: Luna‐Vital Diego , De Mejia Elvira , Dia Vermont , Loarca‐Pina Guadalupe 
Published: June 2021

The aim was to characterize peptides present in non‐digestible fraction (NDF) of common bean cultivars, Azufrado Higuera (AH) and Bayo Madero (BM), produced after pepsin‐pancreatin digestion, determine their antiproliferative action on human colorectal cancer cells and their mechanism of action. Five peptides represented 70% of total protein in both cultivars (GLTSK, LSGNK, GEGSGA, MPACGSS and MTEEY). HCT116 colorectal cancer cell line was most sensitive to bean AH (IC50=0.53 mg/mL), and RKO to BM (IC50=0.51 mg/mL) peptide extracts. Cultivars AH and BM increased significantly the expression of p53 in HCT116 cells by 76% and 68%, respectively. AH modified the expression of cell cycle regulation proteins p21 (64%) and cyclin B1 (‐45%). BM modified the expression of apoptosis‐related proteins BAD (22%); cytC (66%), c‐casp3 (115%); Bax (50%), Bcl‐x (‐41%); Livin (‐32%); Survivin (‐27%) and TNFR1 (‐51%). Moreover, peptide extracts of both cultivars modified the expression of oxidative stress response‐related genes. In HCT116, NRF2‐related antioxidant enzymes were up‐regulated. Glutathione redox‐related enzymes were down‐regulated in RKO, potentially causing reactive oxygen species imbalance. In conclusion, peptides in common bean NDF contributed to the antiproliferative effect on human colorectal cancer cells by modifying molecules involved in cell cycle progression and apoptosis.Grant Funding Source: Supported by University of Illinois‐University of Queretaro research grants
Acute effects of brewed cocoa consumption on sustained attention, motivation to perform work and feelings of anxiety, energy and fatigue (647.1)
Authors: Boolani Ali , Lindheimer Jacob , Loy Bryan , O'Connor Patrick 
Published: June 2021

Acute effects of brewed cocoa and brewed cocoa with 49 mg added caffeine on attention, motivation and mood were examined in a randomized, double‐blinded, crossover trial. Participants (n=23) were low consumers of polyphenols without elevated feelings of energy. Prior to and three times after treatment, a 26‐minute battery was performed assessing motivation, mood and sustained attention (serial subtractions of 3 and 7, the continuous performance task and the Bakan dual task). Treatment X Time ANCOVAs controlling for prior night’s sleep showed that compared to placebo, cocoa reduced overall false alarms and speeded response times on the Bakan secondary task that was completed 31‐47 minutes post‐consumption. Caffeinated cocoa showed: (i) an attenuation of the anxiety provoking effects of cognitive testing found after drinking caffeine alone, and (ii) increased accuracy and reduced omission errors on the primary task of the Bakan test compared to cocoa alone. Brewed cocoa can acutely influence aspects of sustained attention but it has little effect on motivation to perform cognitive tasks or feelings of energy and fatigue. Supplemental caffeine in brewed cocoa can enhance aspects of sustained attention while brewed cocoa can attenuate the anxiety provoking effects found from drinking caffeine alone.Grant Funding Source: The Hershey Company
Overexpression and function of small conductance Ca2+‐sensitive K+ channel (SKCa) isoform 3.1 in HL‐1 myocyte mitochondria (648.12)
Authors: Yang Meiying , Camara Amadou , Bienengraeber Martin , Kwok Wai‐Meng , Stowe David 
Published: June 2021

SKCa agonists protect against cardiac ischemia and reperfusion injury. SKCa channels are present in cell membranes of vascular and cardiac atrial cells, but not in cell membranes of cardiac ventricular cells. Of the 3 known SKCa isoforms, we found SKCa isoform 3 of splice variant 1 (SKCa 3.1) in the inner mitochondrial (m) membrane of guinea pig ventricular cells. To determine its mitochondrial targeting sequence and function, we cloned full length (FL) and truncated SKCa 3.1 cDNA for overexpression in HL‐1 myocytes. SKCa 3.1 localization was examined by immunocytochemistry and western blotting and channel function was evaluated by K+ uptake into HL‐1 myocyte mitochondria with the K+‐selective fluorescent probe PBFI AM. We found that SK3FL‐EGFP, N‐terminus truncated SK3.1 (SK3Δ1‐277‐EGFP), but not C‐terminus (includes calmodulin binding motif) truncated SK3.1 (SK3Δ626‐720‐EGFP), localized into mitochondria when transfected into HL‐1 cells. Overexpression of SK3FL‐EGFP and SK3Δ1‐277‐EGFP, but not SK3Δ626‐720‐EGFP, increased the rate of K+ uptake in HL‐1 cell mitochondria in response to added KCl and CaCl2. K+ uptake by overexpressed SK3FL‐EGFP was sensitive to the SKCa blocker apamin. Our results show that mSKCa 3.1 requires its C‐terminus not only for mitochondrial targeting but also for Ca2+ ‐stimulated K+ uptake into mitochondria.Grant Funding Source: R01 HL 089514
Novel murine model of hepatocyte‐specific human platelet‐derived growth factor alpha expression (649.9)
Authors: Kikuchi Alexander , Lee Jung Min , Monga Satdarshan 
Published: June 2021

Hepatic fibrosis is a common outcome of chronic liver disease from many etiologies and represents a potentially reversible stage of disease progression. Hepatocytes play an important role in this pathologic process through apoptosis, and release of cytokines and growth factors to influence myofibroblast activation and altered proliferation. We have found elevated hepatocyte platelet‐derived growth factor α (PDGFRα) expression in experimental fibrosis and in patients with fibrosis, as well as reduced injury in PDGFRα‐conditional knockout mice. While these findings suggest that PDGFRα upregulation may be involved in hepatic injury, it remains undetermined whether PDGFRα upregulation in hepatocytes plays a compensatory or pathological role. To address this question, we have generated a novel transgenic mouse model expressing human PDGFRA in hepatocytes under the control of an albumin‐promoter. We show that this animal expresses human PDGFRα in whole liver lysates, and overexpresses total PDGFRα but lacks an overt phenotype. In conjunction with previously described human and mouse‐specific monoclonal anti‐PDGFRα antibodies, this mouse strain will allow the investigation of a cell‐specific role of PDGFRα in experimental models of chronic liver injury and regeneration.Grant Funding Source: T32: HL094295 ANGIOPATHY TRAINING GRANT; R01: DK095498 ROLE OF PDGFRALPHA IN LIVER PATHO‐BIOLOGY
Acute high fat diet promotes NF‐κB activation and cardioprotection through circulating adiponectin (652.1)
Authors: Amratia Avni , Haar Lauren , Wani Mashhood , Gonzalez Christopher , Jones Walter 
Published: June 2021

Prolonged high fat diet (HFD) consumption leads to diverse pathologies such as obesity, type II diabetes, atherosclerosis, and myocardial infarction. Paradoxically, studies have shown that short term HFD is cardioprotective against myocardial ischemia and reperfusion (I/R). This response is NF‐κB dependent, a hallmark of preconditioning pathways. This study will address the hypothesis that the adipose‐derived circulating cytokine, adiponectin promotes the activation of NF‐κB in an acute HFD by acting on cardiac receptors and stimulating cardioprotective gene programs.To investigate this in an intact in vivo AdipoQ KO model, infarct size formed in 24h HFD and ctrl chow fed mice were compared. In vitro studies employed HL‐1 cells treated with adipocyte conditioned or ctrl medium to assess a direct response of adiponectin on cardiomyocytes after undergoing a simulated I/R (sim I/R) event in a hypoxic chamber. Cell death was measured by lactate dehydrogenase assay and NF‐κB activation was indirectly measured through the activation downstream NF‐κB ‐dependent genes (VEGF, HIF1a) by qRT‐PCR.Results showed no significant difference in infarct size between the HFD and ctrl chow AdKO mice suggesting the loss of HFD cardioprotection due to loss of adiponectin. Furthermore, treatment in conditioned medium demonstrated significantly reduced cell death and upregulation of NF‐κB‐dependent gene programs after sim I/R in comparison to untreated cells. Our data supports adiponectin as a crucial molecular activator in acute HFD cardioprotection.
Tyrosine phosphorylation of HSC70 may mediate methotrexate resistance in murine L210 leukemia cells (655.5)
Authors: Liu Tuoen , Singh Ratan , Sheridan Peter , Rios Zechary , Bhushan Alok , Lai James , Daniels Christopher 
Published: June 2021

We previously identified and characterized a 66‐68 kDa membrane‐associated, tyrosine phosphorylated protein in murine leukemia L1210 cells as HSC70 which is a methotrexate (MTX) ‐ binding protein. We further characterize the functional role of HSC70 in regulating MTX resistance in L1210 cells. The tyrosine phosphorylation status of the HSC70 in the membrane fraction is different between the parental L1210/0 and cisplatin (CDDP)‐MTX cross resistant L1210/DDP cells. The binding assay with MTX‐agarose beads showed HSC70 in the L1210/DDP cells had less binding ability with MTX compared with L1210/0 cells. In addition, genistein (a tyrosine phosphorylation inhibitor) significantly enhanced the resistance of L1210/0 cells to MTX. In addition, site‐directed mutation studies indicated the importance of tyrosine phosphorylation of HSC70 in regulating its binding to MTX. These studies suggested that tyrosine phosphorylation may contribute to the functions of HSC70 in regulating MTX resistance in L1210 cells.Grant Funding Source: Supported by National Institutes of Health Grant P20 RR16454 and P20 GM103408
Effects of selective dopamine D3 receptor compounds in nonhuman primate models of cocaine and methamphetamine abuse (661.8)
Authors: John William , Green Heather , Newman Amy , Nader Michael 
Published: June 2021

Within the dopaminergic system, the D3R has been shown to mediate many of the behavioral effects of psychostimulants associated with high abuse potential. The present studies examined the effects of D3‐selective partial agonists and D3/D2R antagonists in three models of cocaine and MA abuse in rhesus monkeys. The highly selective D3 partial agonist PG619 and the D3/D2R antagonists buspirone and eticlopride were administered chronically to monkeys self‐administering cocaine and MA (n=3/group) under a food‐drug choice procedure in which complete cocaine and MA dose‐response curves were determined each session. Neither PG 619 (0.1‐3.0 mg/kg, i.v.), buspirone (0.01‐0.3 mg/kg, i.m.) nor eticlopride (0.001‐0.01 mg/kg, i.v.) significantly decreased cocaine or MA choice and intake while buspirone significantly increased the choice of low drug doses. The ability of buspirone to block reinstatement was also examined. Using the choice procedure in which saline was substituted for the self‐administered drug, buspirone (1.0‐1.7 mg/kg, p.o.) was found to block both cocaine‐ and MA‐induced reinstatement. In relation to this, buspirone (0.03‐0.3 mg/kg, i.m.) was also found to be effective in blocking cocaine and MA discrimination in a separate group of monkeys (n=3/group). Overall, these data indicate that while buspirone did not block the reinforcing effects of stimulants when studied under a food‐drug choice paradigm, it was effective in reducing reinstatement and in attenuating the discriminative stimulus effects of cocaine and MA, supporting a potential use for this compound in drug abuse treatment. DA012460Grant Funding Source: Supported by DA012460
l‐Stepholidine is a potent pan‐dopamine receptor antagonist at G protein and β‐arrestin mediated signaling in vitro (662.5)
Authors: Meade Julie , Free R , Miller Nicole , Moritz Amy , Conroy Jennie , Doyle Trevor , Watts Val , Sibley David 
Published: June 2021

Dopamine receptors (DARs) are highly validated drug targets in neuropsychiatric disease and, as such, compounds acting on this receptor family are both scientifically and clinically important. The Chinese herbal extract component l‐stepholidine has been reported to exhibit D2 DAR antagonism and D1 DAR agonist‐like activities in a number of in vivo behavioral analyses. Prior studies have further shown complicated mixed pharmacological responses involving both dopamine and serotonin receptors; however, clear in vitro characterization on individual DAR subtype signaling pathways has not been extensively explored. These data are particularly important given the recent discovery of the potential for signaling bias within the DAR family. In this study we present a broad examination of the biological actions and binding activities of l‐stepholidine on all DAR subtypes. l‐Stepholidine demonstrated relatively high binding affinity for all DAR subtypes, but failed to display any agonist response in either G protein‐ (cAMP assays and GTPγS binding), or β‐arrestin‐ (β‐arrestin translocation assays) mediated signaling at any DAR tested. Furthermore, l‐stepholidine was a full antagonist of dopamine mediated signaling at both D1‐ and D2‐like receptors and did not display signaling bias, thereby demonstrating that it acts as a non‐selective pan‐dopamine receptor antagonist in vitro.Grant Funding Source: NIH Intramural Research Program
CHAC1 degrades glutathione, sensitizing cells to oxidative injury (663.10)
Authors: Crawford Rebecca , Higdon Ashlee , Prescott Eugenia , Mungrue Imran 
Published: June 2021

CHAC1 is a pro‐apoptotic member of the endoplasmic reticulum stress pathway. The yeast homologue (YER163c) is the first known cytosolic protein to actively degrade glutathione (GSH), the most abundant antioxidant in the cell. However, the enzymatic activity of CHAC1 has not been fully characterized in mammalian cells. Herein, we investigate the effects of modulating CHAC1 in vitro on levels of cellular GSH and components of the GSH metabolic pathway in mammalian cells. Additionally, we define a role for CHAC1 in sensitizing cells to oxidative damage following heme treatment.Overexpression of CHAC1 in human and mouse cells leads to a large decrease in total GSH, comparable to treatment with 250uM buthionine sulphoximine, a chemical inhibitor of GSH synthesis. We probed the effects of co‐administration of excess heme, an inducer of intracellular oxidative stress. CHAC1 overexpression increased sensitivity to heme‐mediated toxicity and decreased total GSH. Supplementation with N‐acetyl cysteine, a GSH precursor, alleviated this sensitivity. CHAC1 overexpression and knockdown leads to modulation of the mRNA for the GSH anabolic enzymes as measured by qPCR.Collectively, this evidence suggests that CHAC1 sensitizes cells to oxidative stress due to degradation of GSH and potential feedback in the GSH anabolic pathway.Grant Funding Source: RRC supported by Louisiana Board of Regents Fellowship #LESQSF; INM supported by NIH grant #HL094709
Essential role of adiponectin in ischemic postconditioning cardioprotection and its association with signal transducer and activator of transcription 3 (663.7)
Authors: Li Haobo , Irwin Michael , Xia Zhengyuan 
Published: June 2021

Ischemic postconditioning (IPostC) protects the heart against ischemia reperfusion injury (IRI) by activating signal transducer and activator of transcription 3 (STAT3). Adiponectin (APN), a protein with anti‐IRI property, activates STAT3 (Cell Metab. 2011, 13: 401‐12), but its role in IPostC is unknown. We postulated that APN is essential for IPostC to confer cardioprotection. Wild type (WT) and APN knockout (KO) mice were subjected to 30 min coronary occlusion followed by 120 min reperfusion without or with IPostC achieved by 3 cycles of 10s of ischemia/10s of reperfusion. Cardiac function was assessed by pressure‐volume (PV) loop system. At the end of reperfusion, KO mice displayed larger infarct size and higher plasma creatine kinase‐MB, but lower end‐systolic PV relation (a reliable measure of ventricular function) and dP/dt (all P<0.05, vs. WT). IPostC significantly attenuated all these changes and increased phosphorylations of cardiac STAT3 (at Ser727), Akt (at Ser473) and eNOS in WT but not in KO mice. In cultured H9C2 cells exposed to 60 min hypoxia (H)/120 min reoxygenation (R), gene silencing with APN or STAT3 siRNA all abolished hypoxic postconditioning (HPostC, 3 cycles of 5 min H/5 min R) mediated cell protection. APN supplementation restored HPostC protection in APN siRNA treated but not in STAT3 siRNA treated cells. We conclude that APN is a key protein for IPostC to activate STAT3 to confer protection.Grant Funding Source: Supported by RGC/GRF grant (784011) and NSFC grant (81270899)
Inflow versus outflow ischemia: differentiation and pharmacological characterization using cellular models (663.8)
Authors: Yang Ya , Lai Ted 
Published: June 2021

Ischemia is a hallmark of stroke, myocardial infarction, and many other tissue injuries, and perhaps one of the best modeled human disease in laboratory animals. Nevertheless, it remains difficult to dissect the two main components of ischemia, the disruption of blood inflow versus blood outflow, in animal models. In this study, we attempt to differentiate these two components of tissue ischemia using cellular models. Inflow‐ischemia is induced by exchanging the nutrient‐enriched culture medium with a nutrient‐less extracellular solution, thereby mimicking the loss of nutritious blood supply. Outflow‐ischemia is induced by reducing the volume of the culture medium to allow more efficient accumulation of secretory products including metabolic wastes, thus resembling the disruption of blood outflow during tissue ischemia. Interestingly, while human embryonic kidney (HEK) cells are resistant to inflow‐ischemia, they degenerate abruptly upon outflow‐ischemia. In marked contrast, rat primary cortical neurons are highly susceptible to inflow‐ischemia, but strongly resistant to outflow‐ischemia. Lastly, the mouse endothelioma cells are resistant to either inflow‐ and outflow‐ischemia alone, and degenerated only when simultaneously subjected to both components of ischemia. As proof‐of‐concept, we demonstrated that outflow‐ischemia is transferrable between cell cultures by exchanging their culture media, and is fully reversible by re‐administration of fresh culture medium. Consistent with metabolic acidosis, cell death could be exacerbated and mitigated by decreasing and increasing pH of the culture medium, respectively. Moreover, the inhibitor amiloride prevented outflow‐ischemic death in a dose‐dependent manner. Taken together, the data reported here showed that different cell types are differentially susceptible to different components of ischemia, and further identified amiloride‐sensitive receptors as pharmacological targets against outflow‐ischemia.Grant Funding Source: Supported by the National Science Council of Taiwan
Infusion of cardioplegia with monounsaturated lipids attenuates myocardial injury: involvement of lipid rafts in the protective effect (667.8)
Authors: Maldonado Claudio , Rodriguez‐Alvarez Walter , Khundmiri Syed , Pantalos George , Bauer Philip , Keeling W. Brent , Gray Laman , Perez‐Abadia Gustavo 
Published: June 2021

Myocardial ischemia/reperfusion injury (IRI) can lead to deleterious clinical outcomes following cardiopulmonary bypass (CPB). We developed fusogenic lipid vesicles (FLVs) formulated with monounsaturated lipids (MULs) that modulate endothelial cell (EC) adhesion molecule expression. The purpose of this study was two fold: 1) determine in a swine CPB model the effect of infusing crystalloid cardioplegia (CC)+FLVs in attenuating IRI; and 2) elucidate potential mechanisms by which FLV administration provides protection. We hypothesize that FLVs rapidly incorporate MULs into EC lipid raft microdomains (LRMs) altering their composition, which in turn interferes with EC activation signaling. To evaluate efficacy of FLV treatment swine hearts were infused with CC or with CC+FLVs and underwent 2h of cross‐clamp ischemia. After clamps were removed, cardiac function was evaluated during 2h of reperfusion. To evaluate potential cellular mechanisms of FLV protection, in vitro studies to visualize incorporation of MULs into EC LRMs using advanced fluorescent microscopy techniques were performed by transfecting cells with plasmid expressing green fluorescent protein‐labeled flotillin‐1, and by treating cells with rhodamine‐labeled FLVs. Results from CPB studies indicated that CC+FLVs treatment reduced myocardial injury and improved cardiac stroke work index, and results from in vitro EC studies indicated that incorporation of FLV MULs into membranes was predominantly associated with LRMs. We conclude that FLV induced incorporation of MULs attenuates myocardial injury after CPB, and that the protective effect appears to be associated with the incorporation of MULs into EC LRMs.Grant Funding Source: Supported in part by grants from NIH 1R42HL079855 and EndoProtech, Inc.