bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression and activity of single‐chain recombinant eel follicle stimulating hormone in mammalian cells (792.2)
Authors: Park Chae‐Won , Nanjidsuren Tsevelmaa , Kim Min‐Su , Seo Eun‐Bi , Hong Sun‐Mi , Kang Myung‐Hwa , Kim Dae‐Jung , Min Kwan‐Sik 
Published: June 2021

The glycoprotein hormone family consists of luteinizing hormone (LH), follicle‐stimulating hormone (FSH), and thyroid‐stimulating hormone (TSH), which are secreted by the pituitary gland in all mammalian species, and chorionic gonadotropin (CG), which is secreted by placental trophoblast cells in primates and equids. These hormones consists of non‐covalently associated α and β subunits. Within a species, the amino acid sequence of the α subunit is identical across all glycoprotein hormones and is encoded by a single gene. The αβ dimer is the active form of the hormone, and biological specificity is conferred by the β subunit. Also in fish, the duality of gonadotropin hormone (GTH), GTH‐1 (FSH‐like GTH), and GTH‐II (LH‐like GTH) is found in certain teleost. In this study, in order to understand the fundamental mechanisms involved in teleost reproduction and to proved a broader basis for comparative studt of teleost GTHs, we isolated a cDNA encoding the α and β subunit of GHT‐1 from eel and produced it's recombinant protein from E.coli and CHO cells. To obtain tethered eelFSH, the cDNA encoding the full‐length eel FSH β‐subunit (signal sequence of 22 amino acid residues and the mature protein of 105 amino acid residues) was fused with the mature protein (93 amino acids) of the α‐subunit by overlapping PCR mutagenesis. Subsequently, PCR fragement was inserted into the pCR2.1 clon ing vector and sequenced. After then, it was digested with the EcoRI and SalI enzymes and ligated into the eukaryotic expression vector pcDNA3. The gene was completely sequenced to confirm the presence of the Kozak site and, myc‐tag and to role out the possibility of any PCR erros. Eel FSH protein was expressed in the E. coli system using the pREST expression plasmid vector, and the protein was purified by NI‐NTA column. Eel FSH protein was detected at the predicted molecular weight of 24.3 kDa by SDS‐PAGE in E.coli and 32‐45 kDa in CHO cell lines.Grant Funding Source: This work was supported by NFRDI
MEK kinase 2 regulates integrin signaling to control paxillin ubiquitylation and focal adhesion stability in breast tumor cells (797.3)
Authors: Cuevas Bruce 
Published: June 2021

We have recently shown by gene silencing in invasive breast tumor cells that the kinase MEKK2 is required for xenograft metastasis, and our objective in this work was to determine how MEKK2 influences tumor cell function. We discovered that silencing MEKK2 expression significantly inhibits tumor cell migration, and was associated with a marked increase in the number and size of focal adhesion complexes. We discovered that tumor cell attachment to fibronectin, but not collagen, induces MEKK2 activation and translocalization to focal adhesions. Furthermore, MEKK2 knockdown greatly reduces fibronectin‐induced focal adhesion kinase phosphorylation, suggesting that MEKK2 mediates integrin‐induced signaling. We reveal that MEKK2 associates with the scaffold protein paxillin in an interaction that requires the amino‐terminal leucine‐rich motif (LD1) of paxillin. We demonstrate that the MEKK2/paxillin complex promotes paxillin ubiquitylation and consequent re‐distribution to the cytoplasm, thereby promoting focal adhesion turnover and cell migration. Finally, we show that MEKK2 associates with the ubiquitin ligase RNF5 and promotes paxillin ubiquitylation. Taken together, our results reveal a novel function for MEKK2 as a regulator of ubiquitylation‐dependent paxillin redistribution that promotes focal adhesion turnover and breast tumor cell migration.Grant Funding Source: NIH CA120881
CRTC2 regulates miRNAs and binds to chromatin remodeling factors to control germinal center B cell fate (798.1)
Authors: Najdi Rani , Zambrano Angara , Sherman Mara , Damore Michael , Teitell Michael 
Published: June 2021

When B cells are stimulated by T cell‐dependent antigens during an immune response, they form germinal centers (GCs) to undergo clonal expansion and affinity‐based selection of surface Ig proteins. GC B cells are highly proliferative and misregulation of normal GC processes results in hematologic malignancies. Our lab has shown that inactivation of CRTC2, a CREB transcriptional co‐activator, and the repression of its target genes are required for B cells to exit the GC and differentiate into plasma cells. CRTC2 is inactivated by a signaling pathway triggered by DNA double strand breaks during Ig gene remodeling. This inactivation process ends with the nuclear export of CRTC2, shutting down a gene program to enable GC B cells to differentiate into plasma cells. Despite the significance of this signaling pathway and its CRTC2‐regulated target genes, key details and mechanisms of CRTC2 regulation during B cell differentiation have not been fully explored. Using a bioinformatics‐coupled molecular approach, we identified multiple candidate CRTC2‐target microRNAs that could regulate GC B cell physiology. Our data also reveals that CRTC2 interacts with components of chromatin remodeling complexes at target promoters in GC B cells. Our findings predict that CRTC2, through its control of miRNA expression and interactions with co‐regulatory factors, regulates B cell development and lymphomagenesis.Grant Funding Source: Supported by R01CA90571, R01CA156674, P01GM081621 (all to M.A.T.) and T32‐CA009056 (to R.N.)
Regulation of the epigenetic signature in leukemia by Ikaros (800.5)
Authors: Payne Jonathon , Casiano Carlos , Dovat Elanora , Song Chunhua , Payne Kimberly , Payne Marvin , Dovat Sinisa 
Published: June 2021

Ikaros encodes a DNA binding protein that regulates gene expression via chromatin remodeling and epigenetic mechanisms. The loss of Ikaros activity due to genetic or functional inactivation results in the development of high‐risk leukemia in humans. Our goal is to determine the mechanisms by which Ikaros regulates transcription in human leukemia. Previous studies showed that Ikaros function in leukemia is controlled through its direct phosphorylation by Casein Kinase II (CK2). Treatment of leukemia cells with CK2 inhibitors results in enhanced Ikaros activity, which leads to cessation of cell growth. We have studied the mechanism by which inhibition of CK2 regulates Ikaros‐induced epigenetic changes in leukemia. The human Nalm6 pre‐B cell leukemia was treated with CK2 inhibitor, and the epigenetic signature of the histone modifications H3K4me3 and H3K36me3 were determined using chromatin immunoprecipitation coupled with next generation sequencing (ChIP‐SEQ). Both H4K4me3 and H3K36me3 histone modifications are associated with positive regulation of gene expression. The enrichment of a particular histone modification was confirmed by quantitative chromatin immunoprecipitation (qChIP). Our data demonstrate that the inhibition of CK2 activity in leukemia results in a marked alteration in the epigenetic signature of H3K4me3 and H3K36me3 as compared to untreated cells. This is directly related to altered Ikaros binding to its target genes following CK2 inhibition. Current bioinformatics analysis is directed toward establishing a link between epigenetic modifications and Ikaros binding in leukemia. These results suggest that CK2 and Ikaros regulate gene transcription in leukemia via histone modifications.Grant Funding Source: Supported by NIH R01 HL095120 (SD); NIHR21CA162259 (KP), St. Baldrick's Foundation Awards (SD,KP,CC)
Mitotic kinase CDK1 cooperates with polo kinase to ensure the temporal regulation of kinetochore microtubule dynamics and spindle checkpoint (802.14)
Authors: Yu Huijuan , Zhuang Xiaoxuan , Mo Fei , Fang Guowei , 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, at which mitotic kinase circuitry synergized to ensure the quality control of cell division. Mitotic master kinase Cdk1 controls many aspects of mitotic chromosome behavior and spindle microtubule dynamics to ensure accurate chromosome segregation. Polo kinase 1 (PLK1) is a highly conserved mitotic kinase and activation of PLK1 requires phosphorylation of the T‐loop. Here we show that phosphorylation of PLK1 T‐loop by CDK1 in conserved Thr214 is essential for temporal control of kinetochore microtubule dynamics and spindle checkpoint signaling. Phosphorylates PLK1 at Thr214 suppresses microtubule dynamics and promotes the stabilization of initial kinetochore‐microtubule attachments in prometaphase. Using fluorescence resonance energy transfer‐based reporter of PLK1 activity at the kinetochore and quantitative analysis of native PLK1 substrate phosphorylation, we show that constitutively phosphorylation of Thr214 reduces inter‐kinetochore tension and turnover of kinetochore microtubules, resulting in increased microtubule attachment errors and a checkpoint dependent mitotic arrest. Therefore, we reason that phosphorylation of PLK1 by CDK1 is essential to ensure the temporal regulation of kinetochore microtubule dynamics and spindle checkpoint control.Grant Funding Source: DK56292; CA164133
Investigating proximity‐mediated catalysis by a protein kinase: how docking affects MAPK specificity and processivity (802.16)
Authors: Kaoud Tamer , Dalby Kevin 
Published: June 2021

To understand how MAP kinases phosphorylate their substrates we previously investigated the mechanism of Ets‐1 phosphorylation by ERK2. Using pre‐steady state kinetic approaches we showed that the interaction between ERK2 and Ets‐1 is transient and that turnover is limited by both phosphorylation and ADP dissociation. Here, we investigate the mechanism of phosphorylation of ATF2 by JNK2 using pre‐steady state kinetics and mutagenesis. We find that JNK2 catalysis is limited by dissociation of ATF2 from JNK2 and that this difference in mechanism has significant implications for both the site‐specificity and processivity of the MAPK. We discuss these results in terms of a general mechanism for substrate turnover by protein kinases.Grant Funding Source: Supported by NIH GM59802 and Welch Foundation (F‐1390)
RNA interference of MRCK‐1 in Caenorhabditis elegans to explore oxidative stress responses (802.26)
Authors: Robben Michael , Erickson Patti 
Published: June 2021

The Caenorhabditis elegans myotonic dystrophy kinase‐related, Cdc42 binding kinase (MRCK‐1) is activated by the Rho GTPase CDC‐42 (cell division control protein 42), which contains a conserved redox‐sensitive sequence motif on the phosphoryl binding loop. If reactive oxygen species (ROS) can activate CDC‐42, MRCK‐1 may be required to elicit a downstream response to oxidative stress. To test this, mrck‐1 expression was targeted using two RNA interference (RNAi) constructs in transgenic lines of C. elegans that produce green fluorescent protein (GFP) driven by redox‐sensitive promoters. After heat‐shock and chemically‐induced oxidative stress, GFP expression levels were quantified using epifluorescent microscopy. Preliminary data from heat‐shock protein reporter strain CL2070 (hsp‐16.2:GFP) indicate no significant GFP expression changes in the RNAi‐treated worms before or after heat stress, but further studies with different oxidative stressors and reporter strains are underway. If reduction of mrck‐1 is found to affect oxidative stress responses, a new mechanism for prevention of degenerative diseases and aging could be suggested.Grant Funding Source: Supported by the Henson Undergraduate Research fund.
Identification of Gα12‐specific binding determinants in AKAP‐Lbc (802.8)
Authors: Cavagnini Kyle , Martin Joseph , Brawley Douglas , Meigs Thomas 
Published: June 2021

The Rho family of small GTP‐binding proteins are key regulators of cytoskeletal dynamics and cell growth. Rho activation is triggered by guanine nucleotide exchange factors (RhoGEFs), and a specific subset of these proteins, including leukemia‐associated RhoGEF (LARG), are stimulated by trimeric G protein α subunits of the G12/13 class. This interaction is mediated by a regulator of G protein signaling (RGS) homology domain common to these RhoGEFs; however, a RhoGEF lacking an RGS homology domain, termed AKAP‐Lbc, is also activated by the G12/13 class in a signaling pathway required in cardiac development. To identify AKAP‐Lbc regions involved in Gα12 binding, we compared AKAP‐Lbc to other Gα12 targets and found a C‐terminal region with weak homology to axin. A GST‐fusion of this region co‐precipitated Gα12 in constitutively active and inactive forms, in contrast to the activation‐dependent interaction of Gα12 with LARG. In addition, binding experiments utilizing Gα12 and Gα13 harboring a common epitope tag revealed this AKAP‐Lbc domain as preferential for Gα12 interaction, whereas the RGS homology domain of LARG bound Gα13 with greater affinity than Gα12. These findings reveal a novel, Gα12‐specific mechanism for engagement of a non‐RGS RhoGEF by the G12/13 class of trimeric G proteins.Grant Funding Source: Supported by the GlaxoSmithKline Foundation and Lineberger Comprehensive Cancer Center
An adapted education program to increase adherence to the dietary guidelines for Americans: a feasibility study (807.1)
Authors: McGee Bernestine , Johnson Glenda 
Published: June 2021

The National Institutes of Health (NIH) We Can! (Ways to Enhance Children's Activity & Nutrition) was culturally tailored in order to increase adherence to the Dietary Guidelines for Americans, as the vehicle for preventing childhood overweight and obesity in Louisiana families.We Can! Louisiana is an educational program aimed at promoting adherence to the healthful food and physical activity recommendations set forth in the U.S. Dietary Guidelines for Americans and the resulting effect on reducing weight gain and the risk factors for obesity‐related chronic disease in parents and children in rural Louisiana communities.The education program designed with input from representatives of the target population including time and location of classes and selection of the healthy lifestyle promoter from the community, included topics gathered from the focus groups that were historical, social, cultural, and environmental in nature that influence health behavior of the target population. The We Can! Louisiana weekly parent sessions included food preparation instructions, physical activity and a period for participants to discuss progress and challenges in following recommended child feeding principles, and developing self‐efficacy and social support in meeting the U.S. Dietary Guidelines for Americans recommendations. A gardening component was included to encourage adoption of the U.S Dietary Guidelines for Americans. The biweekly sessions were held with the children to reinforce the parental instructions. Parents and children demonstrated sustained interest in weekly nutrition and physical activities and offered suggestions for making the program interesting.Grant Funding Source: USDA/ARS project #58‐6251‐8‐041
Chefs Move to Schools: a pilot study of the influence of outside chefs on school lunchroom behavior (808.11)
Authors: Hanks Andrew , Just David , Wansink Brian 
Published: June 2021

Objective: Since the announcement of the Chefs Move to School program, little research has been done to document its impact on student behaviors. The objective of this study was to examine how a professional chef’s dish affects student behaviors in school lunchrooms.Methods: A high school in Western New York invited a local chef to use school cafeteria ingredients to create an entrée compliant with school lunch regulations. On a pre‐determined Thursday, the chef agreed to prepare three types of pizzas that would be served with the other standard entrées. In this school, pizza was Thursday’s main entrée. Lunch sales and consumption data were collected for 370 students on days when the standard pizza was served and on the one day when the chef‐created pizza was served. Before‐after analyses were utilized to document differences in lunch sales and consumption.Results: High school students were 18% more likely to select the pizza prepared by the chef (p=0.022). Interestingly, among students who purchased lunch at least once during the study days, there was a 20% increase in the chance these students purchased a school lunch (p<0.001), suggesting an increased interest among these students. In terms of consumption, 68% of students ate their whole entrée and these students ate nearly 17% more vegetables (p=0.02) when the chef’s entrées were served. Moreover, these students also ate 13% more of their snacks (p=0.011) when the ne entrées were offered.Conclusions and Implications: This study provides evidence that guest chefs can increase participation among high school students in the school lunch program. In addition, students may eat a healthier meal, though particular attention should be paid to potentially less nutritious items students enjoy eating with the new dish.
Linking infant iron status, brain function, and looking behavior during a memory task (810.36)
Authors: Hammons Julie , Canfield Richard , Robertson Steven , Haas Jere 
Published: June 2021

No definition of iron deficiency (ID) exists for infants under 12 mo, but young infants (4‐6 mo) are at risk of anemia and depleted iron stores. Nutritional insults during peak brain development (late gestation thru early infancy) may affect long‐term memory. Previous human studies used global cognitive measures, and cannot link ID to impaired memory via specific brain abnormalities. This study examined the links between iron status (hemoglobin (Hb), ferritin, transferrin receptor and body iron), memory function, and brain activity in 40 infants, aged 4‐6 mo. Memory performance was assessed via a Paired Comparison (PC) task and brain function via a similar task with synchronous electroencephalography (EEG). Mixed and logistic regression models were used to estimate the effect (OR) of iron status on 1) amplitude and latency of EEG peaks related to memory; 2) novelty preference and look duration in the PC. Covariates include quality of mother‐infant interactions and infant age, gender, and SES. Nearly 10% of infants were anemic (Hb <110 g/L) and over 70% had mild anemia (Hb 110‐120 g/L). Over 60% of infants showed novelty preference in the PC. This work will inform a definition of ID in infants <12 mo.Grant Funding Source: Supported by the USDA/NIFA (Hatch# 2012‐13‐228) and NSF Graduate Research Fellowship (JEH)
Quality evaluation of noodles produced from wheat, pigeon pea (Cajanus cajan) and carrot flour blend (813.8)
Authors: Banjoko Immaculata , Oyelola Taiwo , Kujore Fowotade , Banjoko Oludotun , Adebowale Folakemi , Ajiboshin Isaac 
Published: June 2021

Noodles are becoming an increasingly popular meal among the young and old in Nigeria and other developing countries of the world. There is therefore a need to utilize some less known crops native to the environment in the production.The noodles were produced from a blend of pigeon pea flour, carrot flour, mixed in different proportion with a constant amount of wheat flour as sample A (100% wheat) as control, sample B (60% wheat, 20% pigeon pea, 20% carrot flour) Sample C (60% wheat, 30% pigeon pea, 10% carrot flour) sample D (60% wheat, 10% pigeon pea, 30% carrot flour) were subjected to the following analysis: proximate composition, functional analysis, cooking qualities and sensory evaluation. The result showed that sample A (control) contained: carbohydrate (80.42%) protein (2.18%) crude fiber (2.0%) ash (2.4%) moisture (8.2%), water absorption capacity (0.9%), swelling capacity (1.01%) and bulk density (0.7%) in comparison with other sample result shows that sample D has the highest amount of carbohydrate (80.23%) close to that of sample A and the least amount of protein (4.27%) but slightly higher than that of sample A, it also has least amount of ash content (0.4%), crude fiber (2.6) and moisture content (9.0%) sample C has the highest amount of protein (13.13%) least amount carbohydrate (51.37%) highest amount of fat (16.6%) much higher than that of sample A highest amount of ash and also the highest amount of moisture. Sample B has the highest amount of crude fiber (2.8%) high amount of protein (11.81%) which is next to sample C high amount of fat (13.4%) moisture (11.6%) carbohydrate (57.59%) and ash (2.8%) next to sample C in value. Sample A is most acceptable of all the samples followed by sample C while Sample D is the least acceptable of the samples which is due to high proportion of carrot flour contained responsible for the colour of the overall acceptance.
Gene‐specific changes in DNA methylation in response to chronic folic acid supplementation in normal weight and obese women of child‐bearing age (817.3)
Authors: Park Hea Jin , Shade Deanna , Hausman Dorothy , Hohos Natalie , Meagher Richard , Kauwell Gail , Smith Alicia , Bailey Lynn 
Published: June 2021

Folate plays an important role in one‐carbon metabolism including DNA methylation. We reported a lower acute pharmacokinetic response to folic acid (FA) in obesity. Both maternal folate deficiency and obesity are associated with increased risk for neural tube defects (NTDs). However, the impact of folate or obesity on DNA methylation of NTD‐related genes remains unclear. We provided 800 ug/d of FA to healthy normal weight (NW; BMI 18.5‐24.9 kg/m2;n = 12) and obese (BMI > 30 kg/m2; n = 6) women (18‐35y) for 8‐wks and determined changes in DNA methylation in CD 16+ cells, isolated from blood using magnetic bead technology. DNA methylation was examined across 2107 CpG sites in 91 NTD‐ and folate pathway‐ related genes using the Infinium Human Methylation450 Bead Chip. For each CpG site, the proportion of DNA methylation that changed over the supplementation was evaluated by fitting a separate linear model. The analysis indicated changes in 57 vs. 101 CpG sites (36 vs. 47 genes) following the supplementation in NW and obese women, respectively. Among them, 68% vs. 76% of the CpG sites (26 vs 37 genes) decreased in methylation. These data suggest that DNA methylation in NTD‐ and folate pathway‐related genes changed in response to FA supplementation and that the changes were different in NW and obese women. Specific pathways influenced by folate status and exhibiting a differential response in obesity are being explored.Grant Funding Source: Funded in part by HATCH #GEO00706, #GEO00707 and the Obesity Initiative at the University of Georgia