bluuebunny/crossref_metadata_2025_split huggingface dataset:
MicroRNA‐124 Negatively Regulates Nuclear Receptor Nur77
Authors: Farmer Alexa , Beard Jordan A , Takwi Apana , Wang Yue‐ming , Chen Taosheng 
Published: November 2023

The nuclear receptor Nur77 is commonly upregulated in adult cancers and has oncogenic functions. Nur77 is an immediate‐early response gene that acts as a transcription factor to promote proliferation and protect the cell from apoptosis. Conversely, Nur77 can translocate to the mitochondria and induce apoptosis upon treatment with various cytotoxic agents. Because Nur77 is upregulated in cancer and may have a role in cancer progression, it is of interest to understand the mechanism controlling its expression. MicroRNAs (miRNAs) are responsible for inhibiting translation of their target genes by binding to the 3′UTR and either degrading the mRNA or preventing it from being translated into protein, thereby making these non‐coding endogenous RNAs vital regulators of every cellular process. Several miRNAs have been predicted to target Nur77; however, strong evidence showing the regulation of Nur77 by any miRNA is lacking. In this study we used a luciferase reporter assay containing the 3′UTR of Nur77 to screen 296 miRNAs and found that miR‐124, the most abundant miRNA in the brain with a role in promoting neuronal differentiation, caused the greatest reduction in luciferase activity. Interestingly, we discovered an inverse relationship in Daoy medulloblastoma cells and undifferentiated granule neuron precursors (GNPs) where Nur77 is upregulated and miR‐124 is downregulated. Further elevating Nur77 levels in Daoy cells by exogenous expression increased proliferation and viability, but knocking down Nur77 via siRNA resulted in the opposite phenotype. Importantly, exogenous expression of miR‐124 reduced Nur77 expression, cell viability, proliferation, and tumor spheroid size in 3D culture. In all, we have discovered miR‐124 to be downregulated in instances of medulloblastoma in which Nur77 is upregulated, resulting in a proliferative state that abets cancer progression. This study provides evidence for increasing miR‐124 expression as a potential therapy for cancers with elevated levels of Nur77.Support or Funding InformationThis work was supported by the American Lebanese Syrian Associated Charities (ALSAC), St. Jude Children's Research Hospital, and the National Institutes of Health [Grants RO1GM086415, RO1GM110034, & P30‐CA21765].
Local Aldosterone Synthesis in Skeletal Muscle
Authors: Chadwick Jessica Ann , Swagger Sarah , Lowe Jeovanna , Rafael‐Fortney Jill 
Published: November 2023

OBJECTIVETo characterize mineralocorticoid receptor (MR) regulation in normal vs. dystrophic skeletal muscle and elucidate the mechanism behind the efficacy of MR antagonists in dystrophic muscle.BACKGROUNDOur lab has recently shown MR is present in skeletal muscle fibers and represents a novel therapeutic target for treating both the skeletal and cardiac muscle pathologies of Duchenne muscular dystrophy (DMD). However, the underlying mechanisms and functional role of MR activation in skeletal muscle is still unclear. Excessive MR activation is known to contribute to the pathogenesis of heart failure, presumably due to increased levels of its natural ligand aldosterone. However, clinical trials have shown that addition of an MR antagonist to standard‐of‐care regimens (ACE inhibitors like lisinopril) improves morbidity and mortality in patients, even when circulating aldosterone levels are within the physiological range. There is now a growing body of evidence that shows aldosterone, which was previously thought to only be synthesized in the adrenal cortex can also be synthesized and metabolized locally in extra‐adrenal tissues, including the heart, particularly in pathological states.DESIGN/METHODSAldosterone synthase (CYP11B2) and other key enzymes in the aldosterone synthesis pathway were tested at the protein level in mouse skeletal muscles and myogenic cultures from normal and dystrophic models, and in normal human myogenic cultures using western blot analysis and immunofluorescence. Sequencing analysis of reverse transcribed PCR (RT‐PCR) of RNA from mouse skeletal muscle and human myotubes was used to confirm expression.RESULTSCYP11B2 is present in mouse skeletal muscle homogenates and up‐regulated in muscles from dystrophic mice (Figure 1). CYP11B2 does not appear to be coming from the skeletal muscle itself; immunofluorescence of dystrophic mouse muscles suggests this enzyme may be present in a subset of immune cells, which are present at much higher levels in dystrophic muscles (Figure 1). We have also found type 2 11‐beta hydroxysteroid dehydrogenase (11β‐HSD2), a key regulator of MR specificity, is present in normal human myotubes and is up‐regulated in dystrophic muscle (Figure 2).CONCLUSIONSCYP11B2 and 11β‐HSD2 levels are significantly increased in dystrophic muscle, suggesting MR activation is in excess of the physiological need. 11β‐HSD2 is expressed within the muscle fibers but CYP11B2 appears to be coming from a population of immune cells up‐regulated in dystrophic tissue due to increased inflammation and muscle degeneration.Support or Funding InformationNational Institutes of Health [R01 NS082868]
Dihydroxyacetone Phosphate Acyltransferase is a Peripheral Membrane Protein
Authors: Chilije Maxwell , Hoffarth Elesha , Bertolesi Gabriel , McFarlane Sarah , Zaremberg Vanina 
Published: November 2023

Dihydroxyacetone phosphate acyltransferase(DHAPAT) catalyzes the initial acylation step in the synthesis of ether basedphospholipids in peroxisomes. DHAPAT adds an acyl group to dihydroxyacetonephosphate, a metabolic intermediate from glycolysis, to yield 1‐acyl‐dihydroxyacetonephosphate (1‐acyl‐DHAP). Ether lipids are abundant in the central nervoussystem and DHAPAT deficiency has been associated with many human disordersincluding cataracts and blindness. We will present results from studies using the tractable model organism Xenopuslaevis that also point to a role of DHAPAT in eye development. Analysis of DHAPAT expression by in situ hybridization indicated it is expressed in the lens epithelium and in the proliferative region of the ciliary marginal zone. Given its relevance and how little is known about DHAPAT, we have initiated biochemical studies aimed at fully characterizing this enzyme in terms of structure, function and regulation. Using a yeast heterologous expression system we have been able to overproduce functional X. laevis DHAPAT. A first purification scheme comprised of subcellular fractionation, protein solubilization with different detergents and ionic strength conditions followed by Ni‐NTA affinity chromatography has been developed. Our studies indicate that X laevis DHAPAT is a peripheral membrane protein that can associate‐dissociate from membranes. We are currently exploiting this novel feature for this kind of acyltransferases in order to setup the conditions to improve its purification with the goal of initiating structural studies.Support or Funding InformationThis work was supported by operating grants from the Natural Sciences and Engineering Research Council of Canada to SM and VZ.
Structure‐function analysis of MntP and other manganese exporter proteins and their contribution to reactive oxygen species survival
Authors: Martinez Eli , Zeinert Rilee , Schmitz Jennifer , Draven Joseph , Waters Lauren 
Published: November 2023

Manganese is an important trace nutrient involved in the detoxification of reactive oxygen species (ROS). Manganese also acts as an enzyme cofactor for superoxide dismutase and other enzymes. In bacteria, manganese and Mn‐SOD allow cells to withstand ROS created from internal metabolic processes and external conditions. Pathogenic bacteria require manganese to remain virulent in environments with reactive oxygen species, such as the host organism. Yet, manganese is toxic at high concentrations, so cells tightly regulate Mn levels. In E. coli, manganese homeostasis is maintained by the Mn importer, MntH, and the Mn exporter, MntP. MntP is a transmembrane exporter protein that we predict to traverse the membrane six times with the N‐ and C‐ termini located in the periplasm. Reporter protein fusions were used to study the orientation of MntP within the membrane. The reporter proteins used were alkaline phosphatase (PhoA) and green fluorescent protein (GFP). We created N‐ and C‐ terminal fusions of PhoA and GFP to MntP using Polymerase Incomplete Primer Extension cloning. Using an alkaline phosphatase assay, activity was seen with the C‐terminal fusion, which was confirmed to be oriented in the periplasm. The ability of the fusion proteins to export manganese was reduced, most notably with the N‐ terminal fusion, which was less than 50% active. Additional work on protein fusions is ongoing.Currently, it is known that the ΔmntP leads to Mn sensitivity in cells, coupled with the accumulation of excess Mn. We investigated the relationship between MntP and reactive oxygen species to further characterize MntP. We have now showed that cells overproducing MntP, in comparison to cells lacking the protein, displayed sensitivity to hydrogen peroxide. This phenotype of MntP links the protein to reactive oxygen species resistance for the first time. Additionally, we have created mutations to MntP to further characterize the function of the protein. The new physiological relationship between MntP and reactive oxygen species is being used to study mutations to MntP. In addition to MntP, we have recently isolated new potential manganese exporter proteins. Mutations to potential manganese exporters, created by site‐directed mutagenesis, are being generated. These studies provides insight into the significance of manganese within bacterial cells. By better understanding MntP and other exporter proteins associated to manganese homeostasis, the virulence of pathogenic bacteria could be reduced.Support or Funding InformationResearch Corporation‐ #22658TRIO grant from the United States Department of Education PR/Award Number P217A120210
Qualitative Methods: An Overview for Anatomy Education Research
Authors: Wasserman Jason Adam 
Published: November 2023

This talk will provide a foundation in qualitative methodology that will 1) Inform the audience about the general uses and processes of qualitative research and 2) Highlight the general utility of qualitative study for research in anatomy and anatomy education. It begins by elucidating the limitations of deductive, quantitative approaches—those that first postulate a hypothesis that is then tested with data. These limitations include problems of operationalizing complex and multifactorial concepts, as well as the problems inherent to the prefabrication of hypotheses. The discussion of these limitations helps to outline the utility of inductive, qualitative approaches, which are then described. In particular, this talk focuses on utilizing methodological techniques that originate with data, rather than hypotheses, and then utilize rigorous narrative analytic schemes to produce sound qualitative findings. The talk concludes by elaborating various kinds of questions, germane to anatomy and anatomy education research, which qualitative inquiries are uniquely positioned to address. These include investigating complex patterns of social and emotional responses to cadaveric dissection and complex and iterative patterns of learning. In short, anatomy education must not only deal with promoting the learning of anatomy proper, but also with deeply emotional reactions to dissection, the social‐psychological development of students over time, issues of professional identity formation. Qualitative approaches hold advantages for studying the complexities and ambiguities inherent in these kinds of phenomena and therefore represent an important and underutilized methodology in anatomy education.Support or Funding InformationNone.
Effects of Helichrysum plicatum extract against Thallium(I) sulphate–induced reproductive damage in male rats
Authors: Omur Ali Dogan , Yildirim Betul Apaydın , Kucukler Sefa 
Published: November 2023

In this study, 24 male Sprague Dawley rats aged eight weeks old and weighted 250–300 gr, were used in this experiment. The animals were obtained from Atatürk University Experimental Research Centre and housed in standard laboratory conditions. Commercial pellet chow and fresh drinking water were available ad libitum. Rats were divided into four groups. I. group (n= 6) referred as control group and physiological saline intraperitonally (IP) to the animals. II. group (n=6) referred as 10‐mg/kg Thallium Sulphate (TS) was injected IP to animals. III. Group (n=6) was referred as TS + HPE, 10‐mg/kg TS injected IP and 100 mg/kg HPE peros to animals. IV. group (n=6) was referred as HPE, 100 mg/kg HPE peros to animals. The treatment was performed for consecutive 8 days. The animals were tranquilized and sacrificed on 9ht day of study. Cauda epididymal semen samples and testes tissues were collected. Routine semen examinations were performed and oxidative stress levels of testicular tissues were assayed. Motility in TS group was significantly lower when compared to control and experimental groups. Head and tail abnormality in TS and HPE+TS group were statistically higher when compared to control and HPE groups. MDA level of TS group was significantly higher than in control and treatment groups. Glutathione peroxidase (GPx) and glutathione (GSH) levels in TS group were lower than the other groups. The lowest SOD level was in TS group while the highest level was in control group. MDA level was higher and CAT level was lower in TS group when compared to other groups.Helichrysum plicatum extract is effective on mentioned spermatological and plasma /testis tissue biochemical parameters against Thallium(I) sulphate in terms of testicular toxicity. There are statistically significant differences between mentioned groups as can be seen in the above.Support or Funding InformationAtaturk University Scientific Research Projects Coordination (Project No: 2013‐251)
Mechanisms of Otoconia Development and Clinical Translation
Authors: Lundberg Yesha 
Published: November 2023

Otoconia are bio‐crystals which overlay the sensory epithelium of the utricle and saccule in the inner ear. The crystals couple mechanic forces to the sensory hair cells in these endorgans, a process essential for us to sense linear acceleration and gravity for the purpose of maintaining bodily balance. Otoconia abnormalities are common and can cause vertigo and imbalance in humans. However, the molecular etiology of these illnesses is unknown, as investigators only began to identify genes important for otoconia formation in recent years. This presentation will summarize recent findings, especially those in the presenter's laboratory. While continued efforts are needed to thoroughly understand the molecular mechanisms underlying otoconia development, it is clear that the process involves a series of temporally and spatially specific events that are tightly coordinated by numerous proteins. Such knowledge will serve as the foundation to uncover the molecular causes of otoconia‐related disorders in humans.Support or Funding InformationNIH R01DC008603 and P20RR0187888
Impact of a Comprehensive Nutrition Program on Infant and Young Child Feeding (IYCF) Practices among Children aged 6 to <24 Months in Rural Malawi
Authors: Hurley Kristen M , Kang Yunhee , Monclus Assumpta Bou , Ruel‐Bergeron Julie , Oemcke Rachel , Wu Lee Shu Fune , Mitra Maithilee , Phuka John , Christian Parul 
Published: November 2023

Given the high prevalence of child stunting, the Government of Malawi, with technical support from the World Food Program, launched a large scale comprehensive nutrition program that provides daily LNS to children 6 to <24 months of age and a behavior change communications (BCC) package promoting optimal breastfeeding, meal frequency, and dietary diversity. The objective of this study was to evaluate the one‐year program impact on caregivers’ IYCF practices in Malawi. The study design was quasi‐experimental with one program district and one comparison district. The program impact was measured using cross‐sectional baseline (January‐March, 2014; n=2,364) and midline (January‐March, 2015; n=2,361) survey data. Estimates of impact over one year were generated using Difference in Difference (DID) and Kernel Propensity Score Matching Difference‐in‐Differences (PSM‐DID) analyses using both baseline and midline data in both districts. Dietary diversity remained low at midline with 83.5% of children across both districts not consuming home foods from 4 or more food groups (out of a possible 7) in the past 24 hours. However, improvements in dietary diversity scores (DDS) trended towards being greater in the program district, as compared to the comparison district (p=0.07). Assuming that the nutrient‐density of the LNS was similar to animal meat, the addition of LNS consumption to the DDS increased it by 0.43‐points (p<0.001), and led to a 7.1% improvement in minimum dietary diversity (MDD) and Minimum Adequate Diet (MAD) in the program district as compared to the comparison district (both p<0.05). These results were attenuated in the PSM‐DID analysis with only differences in the DDS including LNS consumption remaining significant (0.4, p<0.05). The frequency of continued breastfeeding and optimal meal frequency were high at midline and did not differ across districts (p>0.05). Some benefits to IYCF practices were found, especially when LNS was included in the DDS calculation. These findings suggest that in the presence of high food insecurity, IYCF messaging and even high knowledge (which appears to exist in both districts) may be ineffective at improving dietary diversity in infants and young children without the provision of a supplemental food.Support or Funding InformationThis study was funded by Children's Investment Fund Foundation, UK.
Effect of Antioxidants on Vitamins A, E and Carotenoid Stability in Dried Blood Spots
Authors: Arredondo Francisco X , Craft Neal E 
Published: November 2023

Dry blood spot (DBS) collection offers a more practical alternative to serum collection, particularly in remote areas where cold storage is not possible. Unfortunately, many analytes, such as retinol and α‐tocopherol, are sensitive to rapid oxidation as the DBS ages on the card resulting in inaccurate measurements. One report indicated that, from day one post‐collection, retinol values can drop up to 25% and α‐tocopherol up to 50%. (Granado, et al. Quimica Clinica 23:121, 2004). Pretreatment of DBS cards with 0.5% sodium ascorbate as an antioxidant improved the recovery of α‐tocopherol during a single determination in several individuals. (Montalvo‐Velarde et al. Rev. Centro de Inv. (Mex) 7:33, 2006).ObjectiveTo investigate the use of DBS cards pretreated with non‐phenolic and phenolic antioxidants on the stability of vitamin A, α‐tocopherol, and carotenoids eluted at different times.MethodsSodium ascorbate (SA) and ascorbyl palmitate (AP) were the non‐phenolic antioxidants; butylated hydroxyanisole (BHA), 2–6, diterbutyl‐4‐methylphenol (BHT) and terbutyl hydroquinone (TBHQ) were used as phenolic antioxidants. Concentrations ranged from 0.005 to 0.5 % in the treatment solutions. What man 903 Protein‐Saver cards were saturated with the indicated antioxidants and allowed to dry overnight. Cards were spotted with 65 μL of blood collected in sodium heparin from a healthy volunteer.DBS retinol, α‐tocopherol, and carotenoids were estimated via reversed‐phase HPLC at 11, 24, 31, and 64 days post‐collection and storage at −20°C. Matching frozen plasma was processed for reference at each time the DBS were examined.ResultsDBS retinol had stabilized by day 11 with recoveries from 60 to 73 % compared to the liquid plasma values. BHA in the pretreatment solution at 0.05% provided the highest retinol recovery at 73.6% and remained stable through day 64. By day‐11, α‐tocopherol values were between 65–80% of those found in liquid plasma. TBHQ at 0.5% in the pretreatment solutions limited oxidation to a 12% drop from day‐11 compared with day 64. This represents a substantial improvement compared to the 50% drop reported by Granado et. al. Lute in was 72–93% of plasma content by Day‐11 and still above 76% in 0.5% TBHQ by Day‐64. β‐carotene had declined to 33–52% of plasma levels by Day‐11. The highest recovery was 28.4% in 0.5% TBHQ. Interestingly, the least % decrease from Day‐11 to Day‐64 for all analytes was observed for AP. Retinol, α‐tocopherol, and carotenoids in liquid plasma remained stable throughout the study.ConclusionsThe results indicate that incorporation of antioxidants into the DBS collection cards improved the recovery of retinol, α‐tocopherol and carotenoids. TBHQ was the most effective antioxidant overall, but AP minimized the % losses from Day‐11 to Day‐64. It is probable that antioxidant treatment of collection cards would be beneficial for other analytes.
Predication of Glycemic Response of Extruded Pulse Products Using In Vitro Analysis
Authors: Thandapilly Sijo Joseph , Mollard Rebecca , Curran Julianne , Bouchard Danielle R , Jones Peter , Ames Nancy 
Published: November 2023

Whole pulses have been extensively studied for their favourable effects on post‐prandial glycemic control in several acute human studies. Despite the large body of evidence showing the glycemic control benefits of pulse flours and fractions, the optimal dose and combination of pulse flours and fractions that should be integrated into commercial products is unknown. Moreover, the impact of processing, such as extrusion, on glycemic response of pulse products has not been thoroughly examined. Accordingly, the current study used an in vitro model to predict the glycemic response of various pulse products, in order to identify potential formulations for human acute feeding trials. Test products comprised of pulse flours (pea, lentil, bean, chickpea) incorporated into extruded snack products at a rate of 40% (replacing corn ingredients). Pea fractions were added to extruded breakfast cereals both as individual ingredients (fibre vs. protein vs. starch) and in different combinations (e.g. fibre + protein; protein + starch; starch + fibre). Current study results showed that addition of pulse flours into extruded corn snacks led to a lower in vitro glucose release (g glucose/100g sample) over 360 min compared to an all‐corn extruded snack. While, combinations of fractions (fibre + protein and fibre + protein + starch) incorporated into extruded oat cereal had lower in vitro glucose release (g glucose/100g sample) over 360 min compared to all‐oat cereal. Analysis of raw material versus processed products showed that processing methods increased the level of starch damage and in vitro glycemic release. In summary, current study results indicate that incorporation of pulse flour or fractions into commercially processed food products will help to improve the glycemic profile of the products. This study results will aid in choosing the right pulse products for human clinical trials based on their in vitro response. Moreover, it will also provide processors with information on how processing influences the nutritional composition and the resulting glycemic response.Support or Funding InformationFunding from Alberta Pulse Growers and Saskatchewan Pulse Growers