bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evaluation of cytotoxicity of CdS/maltodextin nanoparticles bioconjugated with Cry protein in hepatic cells (780.1)
Authors: Gutiérrez Ivonne , Reyes‐Esparza Jorge , Rodriguez‐Fragoso Patricia , Rodriguez‐Fragoso Lourdes 
Published: June 2021

To evaluate of cytotoxicity of CdS/maltodextrin nanoparticles (CdS/MD) bioconjugated with Cry protein in hepatic cells. Previous studies have shown that CdS/MD nanoparticles are innocuous in several cell lines at low concentrations. For this study, the CdS/MD nanoparticles and bioconjugate were analyzed by fluorescence microscopy and electron microscopy. HepG2 cells were treated with CdS/MD alone and bioconjugated with Cry protein. We performed a dose‐response curve of CdS/MD and CdS/MD‐Cry protein [0.01, 0.1, 1‐9 μg/L] and cell viability was measured. The type of cell death was identified by fluorescence microscopy. It was found that the bioconjugate was 20 times bigger than the CdS/MD nanoparticles. It was shown that nanoparticles and the bioconjugate emit green fluorescence in cells HepG2. In cells treated with NPs was observed fully fluorescence, while those treated with the bioconjugate only was observed fluorescence in the cell membrane. The cytotoxicity assay showed that the nanoparticles and Cry protein were non‐toxic at low concentrations (<2μg/L). At low concentrations the bioconjugate did not produce toxicity in HepG2 cells (<0.1 μg/L). The Cry protein produced apoptosis while the bioconjugate produced necrosis. In conclusion, at low concentrations the bioconjugate did not produce toxicity in HepG2 cells.Grant Funding Source: CONACYT
Rhamnetin and cirsiliol regulate radioresistance and EMT by suppression of Notch‐1 expression in NSCLC (794.1)
Authors: Kang JiHoon , Seo HyunJeong , Park Kang Hyun , Youn BuHyun 
Published: June 2021

Despite optimal radiotherapy, chemotherapy and/or surgery, the majority of patients with NSCLC fail treatment. Radioresistance is a major cause of decreasing the efficiency of radiotherapy. To understand its mechanisms, we focused on radiation‐induced Notch‐1 signaling pathway. In this study, we investigated the use of Notch‐1‐regulating flavonoids, rhamnetin and cirsiliol, as novel therapeutic drugs to regulate radiosensitivity in NSCLC. Treatment with two compounds reduced the proliferation of NSCLC cells through the suppression of radiation‐induced Notch‐1 expression. Both compounds increased the expression of miR‐34a in a p53‐dependent manner, leading to inhibition of Notch‐1 expression. Consequently, reduced Notch‐1 expression promoted apoptosis through significant down‐regulation of NF‐κB pathway, resulting in a radiosensitizing effect on NSCLC cells. EMT induced by radiation was also notably attenuated by two compounds. Moreover, an in vivo xenograft mouse model confirmed the radiosensitizing effects of both flavonoids as we observed in vitro. Altogether, our findings provided evidence that rhamnetin and cirsiliol can act as novel radiosensitizers via targeting Notch‐1 expression.Grant Funding Source: Supported by the National R&D Program for Cancer Control, Ministry for Health and Welfare(1320100)
Mitotic kinesin CENP‐E cooperates with microtubule‐bundling protein PRC1 in temporal control of central spindle assembly (801.2)
Authors: Liu Xing , Yao Phil , Wang Wenwen , Qin Bo , Dou Zhen , Ding Xia , Yao Xuebiao 
Published: June 2021

Chromosome movements during mitosis are orchestrated primarily by the interaction of spindle microtubules with the kinetochore, the site for attachment of spindle microtubules to the centromere. Kinetochore‐associated mitotic kinesin CENP‐E plays key roles in chromosome congression and spindle checkpoint satisfaction. We have recently identified and characterized syntelin, a novel selective CENP‐E inhibitor (Ding et al., 2010. Cell Res. 20, 1386‐1390). Cells treated with syntelin progress through interphase, enter mitosis normally with a bipolar spindle and lagging chromosomes around the poles. Syntelin is an allosteric inhibitor which tightens CENP‐E‐microtubule interaction by slowing inorganic phosphate release. To delineate the role of CENP‐E in reorganization of interpolar microtubules into an organized central spindle, metaphase synchronized cells were exposed to syntelin and other mitotic motor inhibitors. Syntelin does not perturb interpolar microtubule assembly but abrogates the anti‐parallel microtubule bundle formation. Real‐time image shows that CENP‐E inhibited cells undergo central spindle splitting and exhibits chromosome instability phenotypes. Interestingly, inhibition of CENP‐E did not alter the interaction between CENP‐E and PRC1 but perturbed temporal assembly of PRC1 to the midzone. Surprisingly, inhibition of CENP‐E perturbs the temporal control of PRC1 dephosphorylation which led to a persistent phosphorylation of PRC1 and an inhibition of central spindle assembly. These findings reveal a previously uncharacterized role of CENP‐E motor in temporal control of central spindle assembly.Grant Funding Source: DK56292; CA164133
Molecular mechanism of Raf kinase inhibitory protein regulation by cAMP‐dependent protein kinase (802.25)
Authors: Ong Colin , Masterson Larry , Kim Jonggul , Gomes Suzana , Veglia Gianluigi , Rosner Marsha 
Published: June 2021

Raf Kinase Inhibitory Protein (RKIP), a 21 kDa protein and member of phosphatidylethanolamine binding protein (PEBP) family, regulates GPCR, MAPK, and NFκB pathways and is a metastasis suppressor. Although, kinases such as ERK2 and PKCα have been shown to phosphorylate RKIP, structural data on the protein kinase‐RKIP interaction are still lacking. Here we report that another kinase, PKA, phosphorylates RKIP at residue 51, triggering a feedback loop, and we determine the structural model of RKIP‐PKA catalytic subunit. Using different biophysical approaches including Nuclear Magnetic Resonance (NMR), Paramagnetic Relaxation Enhancements (PREs) and molecular modeling, we identify the binding interface between RKIP and PKA catalytic subunit. Chemical shift mapping and site directed mutagenesis studies revealed residues within RKIP that are critical to the catalysis and binding mediated by PKA catalytic subunit. As the first description of PKA binding to a full‐length physiological substrate, these results have wide applicability to the mechanism of PKA catalysisGrant Funding Source: Supported by NIH grant GM087630 to Marsha Rich Rosner, NIH grant GM100310 to Gianluigi Veglia
The small GTPase Rab5 controls cell morphology, sexual development, ion‐stress response and vacuolar formation in fission yeast (802.29)
Authors: Miyamoto Masaaki , Tsukamoto Yuta , Katayama Chisako , Shinohara Miki , Shinohara Akira , Maekawa Shohei 
Published: June 2021

Vesicle transport is important for cell functions. Rab family GTPases play a critical role in inner‐membrane transport. In mammalian cells, Rab5 is reported to be involved in the regulation of endosome formation, phagocytosis and chromosome alignment. In order to examine the role of the fission yeast Rab5 homologue Ypt5, we used a point mutant allele and analyzed. Mutant cells displayed abnormal phenotypes in cell morphology, mating, sporulation, endocytosis, vacuole fusion and responses to ion stress. Our data suggest that fission yeast Rab5 is involved in the regulation of various types of cellular functions and regulate cell morphology via actin cytoskeletons.Grant Funding Source: Supported by Kobe Univ. and Osaka Univ.
Analysis of class‐distinctive Gα13 mutants to identify regions critical for serum response element‐mediated transcriptional activation (802.9)
Authors: Tagliatela Alicia , Hamilton Tyler , Jones Alan , Meigs Thomas 
Published: June 2021

Guanine‐nucleotide binding proteins (G proteins) are involved in transmitting signals from cell surface receptors to downstream pathways. The G12/13 class of G proteins includes Gα12 and Gα13, which diverged from a single ancestral G12/13 protein during evolution. These proteins drive proliferation and migration in certain cell types, and also regulate cytoskeleton rearrangements and other cellular events. In order to identify key signaling determinants that evolved in the G12/13 class, we engineered mutants in which class‐distinctive residues in Gα13 were substituted for residues conserved at the same structural position in the non‐G12/13 classes of Gα proteins. These mutants were transfected into human kidney cells, and serum response element (SRE) activation assays identified several substitutions in Gα12 that hindered SRE activation. Surprisingly, the corresponding mutations in Gα13 showed no significant reduction in this serum response. Furthermore, we engineered chimeras in which Gα12 and Gα13 incorporated regions from the fruit fly G12/13 homolog, Concertina, and found a C‐terminal region for which substitution of Concertina sequence abolished Gα12 signaling through SRE yet was fully tolerated in Gα13. These findings suggest Gα12 and Gα13 utilize distinct structural mechanisms for engaging the pathway leading to SRE‐mediated transcription.Grant Funding Source: Supported by the GlaxoSmithKline Foundation and Lineberger Comprehensive Cancer Center
Relationship between apparent in vivo occupancy at the dopamine transporter and cocaine‐like discriminative stimulus effects of N‐substituted benztropine analogs in mice (803.4)
Authors: Wilkinson Derek , Cohen Marika , Newman Amy , Katz Jonathan 
Published: June 2021

Standard inhibitors of the dopamine transporter (DAT) have cocaine‐like subjective effects; however, several atypical DAT inhibitors that bind the DAT with high affinity, such as benztropine (BZT) analogs, do not fully substitute for cocaine in animals trained to discriminate cocaine injections.The lack of cocaine‐like effects may be related to a slow rate of association to the DAT. Several N‐substituted BZT analogs (R‐2‐amino‐3‐methyl‐n‐butyl; 2‐aminoethyl; 2‐ethyl‐3‐indole; and methylcyclopropyl) were reported to have a relatively fast onset of behavioral effects, but it is unclear if these effects were related to DAT activity. Therefore, the present study examined the in vivo binding to DAT of the above BZT analogs and their cocaine‐like discriminative stimulus effects in mice. None of the BZT analogs fully substituted for cocaine discriminative effects despite the testing of doses that produced in vivo DAT occupancy greater than the ~13% occupancy produced by 10 mg/kg training dose of cocaine. The BZT analogs produced displacement of [I125]RTI‐121 from striatum at maximal rates from 0.86 to 1.28 %/min compared to 1.23%/min for cocaine, indicating a relatively fast apparent rate of DAT association. The results of the present study suggest that DAT occupancy, or rate of association, alone cannot account for the lack of cocaine‐like discriminative effects found with these N‐substituted BZT analogs.Grant Funding Source: NIH Intramural Research Program
Identification of the attachment site for the cocaine analog [125I]JHC 2‐48 on the dopamine transporter (803.7)
Authors: Dahal Rejwi , Akula Bala Pramod , Sharma Babita , Cha Joo , Cao Jianjing , Boateng Comfort , Lever John , Newman Amy , Foster James , Henry L. , Vaughan Roxanne 
Published: June 2021

Cocaine is a highly addictive drug that targets the presynaptic dopamine transporter (DAT) and blocks the reuptake of dopamine resulting in heightened dopaminergic neurotransmission. The DAT is a 12 transmembrane (TM) spanning protein with residues from TMs 1, 3, 6, and 8 comprising the substrate and putative inhibitor binding sites; although recent studies indicate the presence of an inhibitor binding site in the external vestibule located above the substrate‐binding site. Despite decades of research, the binding site of cocaine has yet to be fully characterized. To address this question, we are mapping the attachment sites of irreversible cocaine analogs, [125I]MFZ 2‐24, [125I] RTI 82, and [125I]JHC 2‐48, using insight gained from in silico docking with DAT comparative models. These compounds have a cocaine pharmacophore with analog specific azido group that adducts to the protein. To determine the adduction site of [125I]JHC 2‐48 we are using methionine substitution mutagenesis to generate custom protease sites for cleavage by cyanogen bromide (CNBr). The fragments generated from the CNBr proteolysis of [125I]JHC 2‐48 labeled DAT were visualized via SDS‐PAGE and autoradiography. The preliminary results obtained thus far have determined the [125I]JHC 2‐48 attachment site to be C‐terminal to TM7. Peptide map analyses of other cocaine analogs, [125I]MFZ 2‐24 and [125I] RTI 82, have determined [125I]MFZ 2‐24 adducts to TM1 and [125I]RTI 82 adducts to TM6. Taken together, our data strongly indicate cocaine binds near the substrate binding site.Grant Funding Source: Supported by: DA027845 and NIDA‐IRP
Unanticipated weight gain during a dietary intervention designed to improve iron status (804.20)
Authors: Chen Vivi , Luna Sarah , Lung'aho Mercy , Gahutu Jean Bosco , Haas Jere 
Published: June 2021

Objective: To quantify the effect on weight gain of a 4.5 months feeding trial intended to replace the subjects’ regular meals.Methods: 145 Rwandan women (18‐26 years) in a double‐blind feeding trial were randomized to receive either iron‐biofortified beans or traditional beans daily for 4.5 months. During the feeding period, the subjects were served 175 g of beans per meal, for lunch and dinner, plus ad libitum consumption of side dishes; they were allowed to consume more beans if desired. Blood biomarkers and anthropometry were measured before and after the feeding period. General linear models were used to evaluate the effect of the intervention on weight gain.Results: On average, the women consumed 200 g of beans (358 kcals) per meal. After the 4.5 months feeding period, 25% of the women gained 蠅 3 kg in body weight. Both the control and the treatment group consumed the same amount of beans per meal, with the treatment group having a significantly (p>.001) greater increase in iron status. The more beans consumed per meal, the more weight the subjects gained (F= 3.94, p=.05), regardless of iron intake. Women in the upper quartile of the BMI distribution (BMI > 24.2) gained the most weight (F=3.09, p=0.03).Conclusion: Providing lunch and dinner in a feeding trial to improve micronutrient status resulted in a net addition of calories and subsequent weight gain, especially in women with higher BMI.Grant Funding Source: Supported by HarvestPlus
Glycemic load is positively associated with diastolic blood pressure levels during pregnancy: preliminary results from a prospective cohort study in Rio de Janeiro, Brazil (810.11)
Authors: Vilela Ana Amélia , Mendes Roberta , Franco‐Sena Ana Beatriz , Farias Dayana , Rebelo Fernanda , Rodriguez Camila , Kac Gilberto 
Published: June 2021

Objective: To evaluate the association between dietary GL and systolic (SBP) and diastolic blood pressure (DBP) during healthy pregnancy. Methods: Prospective cohort of 155 healthy pregnant women followed at 6‐13th, 20‐26th and 30‐36th gestational weeks. Dietary GL was calculated from glycemic index, carbohydrate content, and frequency intake of selected foods, obtained from food frequency questionnaire applied in the 3rd trimester having the whole pregnancy as the time frame. SBP and DBP throughout pregnancy were the outcomes. Statistical analysis comprised multiple longitudinal linear mixed‐effect models adjusted for obstetric, socioeconomic, dietetic, and biomarker variables. Results: Mean SBP was 109.7, 107.9, and 111.3 (mmHg) and DBP was 66.6, 64.3, and 67.2 (mmHg) for the 1st, 2sd and 3rd trimester, respectively, and didn’t varied according to tertile of GL in the 3rd trimester (p>0.05). GL was not associated with SBP in the multiple model (β=0.237; 95%CI=‐0.16‐0.64; p=0.245). GL was positively associated with DBP (β=0.387; 95%CI=0.08‐0.69; p=0.015) after adjusting for confounders such as per‐capita family income, skin color, BMI, and calories. Conclusion: Dietary GL was positively associated with DBP changes during pregnancy.Grant Funding Source: Supported by: FAPERJ, CNPq, CAPES
Three statistical approaches to the estimation of intervention effect on a randomized cluster trial to reduce anemia and iron deficiency in children aged 12‐30 months (810.27)
Authors: Mendez Gomez Humaran Ignacio , Shamah Levy Teresa , Mendez Ramirez Ignacio 
Published: June 2021

A double‐blinded, group‐randomized trial was conducted in 12 milk distribution clusters. After a randomization device, 7 clusters were assigned to consume iron‐fortified an 5 non‐fortified milk. The objective of the research, is to assess the effectiveness of a large‐scale program that distributes subsidized iron‐fortified milk in Mexico on anemia and iron deficiency in children aged 12‐30 mo. Randomized cluster trials, are experimental studies that attempt to balance confounding factors that affect outcomes across the study groups controlling for possible hidden bias, but randomization is never completely successful.The use of statistical methods to control variability in the study groups in order to make meaningful comparisons and describe characteristics of the groups being compared in order to find a reasonable amount of overlap is encouraged.In this work we compare three statistical methods to establish effect estimates and control variability in the studygroups. First, it is common the use generalized linear models with several covariates to establish corrected effect estimates. Second, a simplest approach is the use cluster analysis to generate multivariate post stratums and use them in those models to control variability among groups. Finally, the use of propensity score matching procedures is useful to estimate treatment effects and approximated bias estimates. All three methods present different ways to address the roll of covariates in the understanding of the whole phenomena.
Qualitative validation and construction instrument for measuring well‐being perception associated with water consumption (810.32)
Authors: Espinosa Juan , Monterrubio Eric , Castañeda Sandra , Barquera Simon 
Published: June 2021

An instrument for assessing well‐being perception associated with water consumption (IPBA) in adults for low socioeconomic status in Cuernavaca, Mexico was constructed and validated. Focus groups were made as well as semi‐structured interviews including adult men and women (21‐59 years old) who are sedentary. Two groups of people with a different consume of water were formed: the ones who consume a lot of water (> 2.0L total liquids (TL) and plain water (PW) < 1L) and the ones who hardly consume any water (< 1.5L of TL and PW < 0.3L). Guidance was used to investigate the meaning of well‐being by PW consumption from the knowledge, attitudes, beliefs and perceptions of everyday life. Internal consistency was assessed by means of Cronbach´s Alpha (CA). Sessions were leaded by expert psychologists and were audio recorded under the oral consent of participants.Items were created from keywords and symptoms related to hydration. IPBA was adjusted by pilot testing; was revised by experts on hydration and sent to judges for validating facie and construct. A total of 55 items were chosen; such items reference perception of well‐being in the last 7 days and uses a liker‐type answer scale (Always, almost always, sometimes, almost never, never). The IPBA identifies two dimensions (Physical well‐being and mental well‐being) with a CA of 0.83 and 0.91 respectively. In a subsequent phase exploratory and confirmatory factors analyses are performed.IPBA is the first proposal of instrument for measuring well‐being perception related to hydration. The construction of items was based on empirical evidence about how the target population senses well‐being related to hydration.Grant Funding Source: Sponsored/funded by Instituto Danone de Mexico
Identification and functional characterization of an alternative 5’ exon of the sodium dependent ascorbic acid transporter SLC23A1 (818.5)
Authors: Amir Shaghaghi Mandana , Yurkova Natalia , Tu Hongbin , Levine Mark , Eck Peter 
Published: June 2021

Vitamin C, an essential micronutrient and a critical cofactor for several intracellular enzymatic reactions, is actively transported across epithelial barriers through the sodium‐dependent ascorbic acid transporter 1 (SLC23A1). The SLC23A1 gene encodes SLC23A1 transporter and is located on human chromosome 5q31.2 (138702885‐138719039 compl.; NC_000005.9). Previously, the SLC23A1 transcript has been described as containing 15 exons, stretching over 16 kilobases; however, no alternative splice variant has been described yet. Variations in SLC23A1 gene would impact vitamin C dietary requirements and/or susceptibility to common complex diseases. We have, therefore, evaluated the existence of additional SLC23A1 transcripts in silico and examined the function of new transcript through Xenopus laevis oocytes injection. Here, we characterized a novel alternative first exon encoding a SLC23A1 isoform which is exclusive to human and is expressed in the small intestine. The novel SLC23A1 protein isoform adds 36 N‐terminal amino acids but does not alter transmembrane topology or intracellular location. The novel SLC23A1 isoform is located on the plasma membrane and mediates very low ascorbic acid transport when expressed in Xenopus laevis oocytes. This is the first report of an alternative splice variant in human SLC23A1 locus and confirmed an extension of about 1kb further 5’ than previously described gene.Grant Funding Source: This work was funded through Peter Eck’s Canada Research Chair in Nutrigenomics award and in part b
Leucine activation of mTORC1 in skeletal muscle cells is inhibited by the lysosomotropic agent chloroquine (820.2)
Authors: Borack Michael , Rasmussen Blake 
Published: June 2021

Recent work in human embryonic kidney cells has uncovered many of the cellular mechanisms underlying amino acid activation of the mTORC1 signaling pathway. It is now known that mTORC1 translocation to the lysosomal surface is required for the amino acid activation of mTORC1 signaling. It is not known if the lysosome is required for amino acid activation of mTORC1 in skeletal muscle cells. To test this hypothesis, we exposed C2C12 muscle cells (that were plated and differentiated into myotubes for 7‐8 days) to both leucine and a potent lysosomal inhibitor (chloroquine). Cells were placed into three groups: control (1), leucine only (2) and leucine+chloroquine (3). The first two groups were taken out of media and starved in a Hepes buffer for 9hrs. Group 3 was starved for 8hrs followed by a 1hr starve in 2000ug/ml chloroquine in the same Hepes buffer. Groups 2 and 3 were given 1mM leucine for ten minutes before collection. Western blotting was used to determine phosphorylation of mTOR and its downstream targets. The leucine only group showed a significant increase in mTOR and p70s6k phosphorylation compared to control and leucine+chloroquine groups (p<0.05). It appears that disruption of lysosomal function by chloroquine blocks leucine activation of the mTORC1 signaling pathway in skeletal muscle cells. Future work will determine the role of the lysosome for amino acid activation of mTORC1 in vivo.Grant Funding Source: Supported by NIH AR049877