bluuebunny/crossref_metadata_2025_split huggingface dataset:
In good taste: An investigation of lactate metabolons in the mouse tongue
Authors: Ochrietor Judith D , Zahir Haaris , Castillo Luz , Seddeik Rita , Gelsleichter Jim D 
Published: November 2023

Recent studies of sensory receptor tissues in the mouse suggest the importance of lactate metabolons within those tissues for proper function. A lactate metabolon consists of a monocarboxylate transporter (MCT) and a member of the immunoglobulin superfamily, such as Basigin gene products. In the neural retina, a lactate metabolon consisting of Basigin gene products and MCT‐1 and MCT‐4, has been deemed necessary for photoreceptor maturation and function. Basigin‐null mice do not express these metabolons and are blind from the time of eye opening. Basigin and MCT‐1 are also expressed on the myelin of olfactory nerve fibers, presumably to form a lactate metabolon required to support saltatory conduction. Basigin‐null mice have olfactory deficits, in that they do not respond to the smell of acetic acid as normal mice do. The absence of the metabolon likely accounts for the slower response rates observed in the null animals. The purpose of the present study was to investigate the presence of a lactate metabolon consisting of Basigin gene products and MCTs in the mouse tongue. Tongues were obtained from mice using an accepted protocol for use in immunohistochemical and quantitative RT‐PCR analyses. It was determined that Basigin is expressed on the mouse tongue epithelium, but not in the underlying muscle. No significant expression of MCT‐1 or MCT‐4 was detected in the tissue; however, MCT‐2 expression was observed within the taste buds. The data suggest that a lactate metabolon like those associated with olfactory nerve fibers and in the neural retina is not present in the mouse tongue.Support or Funding InformationUniversity of North Florida Office of Undergraduate Studies and the Office of Academic Affairs
Exploration of Plasmodium Kinases
Authors: Eubanks Amber L. , Derbyshire Emily R. 
Published: November 2023

Malaria is a neglected tropical disease that affects millions of people worldwide. The World Health Organization reported almost 600,000 deaths in 2013 alone, with most of these among children and pregnant women. Malaria is caused by protozoan parasites from the genus Plasmodium and is transmitted to human hosts through Anopheles mosquitoes. Eradication of this devastating disease is complicated by the complex life cycle of the parasite and increasing drug resistance to first‐line treatments. In order to combat Plasmodium infections and drug‐resistance, a deeper understanding of the parasite biology is required such that better therapeutic targets can be elucidated. Protein kinases are becoming increasingly popular as druggable targets in many diseases due to their essential function in eukaryotic organisms. Unfortunately relatively little is known about the malaria kinome. More than 3% of the Plasmodium parasite proteome is made up of kinases and offers a wide array of targets. The goal of our work is to biochemically characterize malaria protein kinases that are involved in key biosynthesis and signaling pathways. These efforts will allow further biological understanding of parasite biology while also providing potential therapeutic targets. Screening compounds against these kinases will also aid in the discovery of small molecule probes and antimalarial agents.Support or Funding InformationNIHDuke University Chemistry Department
Expression profile of Monocarboxylate Transporters and Basigin gene products in the mouse cornea
Authors: Maniccia Randall G. , Fong Joseph D , Ochrietor Judith D 
Published: November 2023

Monocarboxylate transporters (MCTs) transport lactate, pyruvate, and ketone bodies. Expression of MCTs has been linked to the expression of Basigin gene products, to form a lactate metabolon, that has been determined to be essential for proper photoreceptor function in the neural retina, and hence vision. Since the cornea is the outer, protective portion of the eye, this research group was curious if a similar lactate metabolon is present in that tissue. The group is also curious about the role of MCTs in the wound healing process. Therefore, the purpose of the present study was to determine whether Basigin gene products and MCTs found within the neural retina are also expressed in the corneal epithelium of mice. Corneas were isolated from mice using an accepted protocol and pooled so that detergent‐solubilized proteins and RNA could be isolated from epithelial cells. ELISA analyses and quantitative reverse transcriptase polymerase chain reaction (q‐RT‐PCR), were performed to test for the presence of Basigin gene products, MCT1, MCT2, and MCT4. It was determined via ELISA that Basigin gene products, MCT2, and MCT4 were found within the corneal epithelium; however, MCT1 was not detected. Analyses of RNA from corneal epithelial cells indicate that the ubiquitously expressed short form Basigin is present in the tissue, but the photoreceptor‐specific long form Basigin‐2 was not. In addition, no MCT1 transcript expression was observed in the cornea sample. These data suggest that a lactate metabolon may be present in mouse cornea. In addition, the data are consistent with data obtained by other laboratories using human and rabbit corneas, with the exception of MCT1, which was found in those other species. Future studies will determine the role played by the lactate metabolon during wound healing of the mouse cornea.Support or Funding InformationUniversity of North Florida Office of Undergraduate Studies and the Office of Academic Affairs
High Fructose Corn Syrup‐55 Alters Hepatic Metabolism to Increase Risk of Non‐Alcoholic Fatty Liver Disease in Female Sprague‐Dawley Rats
Authors: Lateef Sundus , Mock Kaitlin , Benedito Vagner , Tou Janet 
Published: November 2023

Studies have reported that high fructose intake promotes de novo lipogenesis (DNL). In turn, DNL stimulation can promote non‐alcoholic fatty liver disease (NAFLD), a metabolic condition that can progress to cirrhosis and liver failure. A frequent alternative to sucrose in calorically sweetened beverages, high fructose corn syrup‐55 (HFCS‐55) contains a higher percentage of free fructose (55%) compared to sucrose (50%, all of which is combined with glucose as a disaccharide). The objective of this study was to determine the effect of HFCS‐55 sweetened solution intake on hepatic lipid metabolism and risk of developing NAFLD in a rodent model. Female Sprague‐Dawley rats (age 28 days) were randomly assigned (n=7/group) to one of four treatment groups consisting of distilled deionized water (ddH2O) or ddH2O sweetened with sucrose, fructose or HFCS‐55 for 8 weeks. Caloric sweeteners were administered at a concentration of 13% w/v, the level typically found in sugar‐sweetened beverages. Gene expression of key transcription factors and enzymes involved in DNL including sterol regulatory element‐binding protein 1c (SREBP‐1c), fatty acid synthase (FAS), and stearoyl‐CoA desaturase 1 (SCD1) as well as peroxisome proliferator‐activated receptor alpha (PPARα), a transcription factor activating lipolysis, were measured using real‐time quantitative polymerase chain reactions (RT‐qPCR). Liver fatty acid composition was determined using gas chromatography and serum clinical markers were measured by enzymatic assays. Expression of SREBP‐1c (P=0.02) and FAS (P=0.001), molecular markers of DNL, were up‐regulated in all caloric sugar treatment groups compared to the ddH2O group. Transcript abundance of SCD1, which is involved in synthesizing palmitoleate (C16:1n‐7) and oleate (C18:1n‐9) from palmitate (C16:0) and stearate (C18:0), was also induced in all caloric sugar treatment groups compared to the ddH2O group (P=0.001). Hepatic palmitoleate content was higher in the sucrose and HFCS‐55 groups compared to the ddH2O group (P<0.001), and hepatic oleate content was highest in the HFCS‐55 group (P<0.001). PPARα expression was lower in the sucrose and HFCS‐55 groups (P=0.002). Altogether, we show evidence that HFCS‐55 intake had a greater effect on fatty acid metabolism than sucrose by promoting DNL, which was not compensated for by increased lipolysis. Rats drinking HFCS‐55 solution had highest hepatic triglyceride content (P=0.03) among all treatment groups and higher low density lipoprotein (P=0.02) than rats drinking ddH2O, indicating NAFLD. Thus, HFCS‐55 intake in healthy growing rats resulted in higher risk of NAFLD indicated by hepatic lipid accumulation and dyslipidemia.Support or Funding InformationThis research was sponsored in part by the West Virginia Research Challenge Fund through a grant from the Davis College of Agriculture, West Virginia University.
Contribution of Poly‐unsaturated Fatty Acids on Cerebral Neurobiology: An Integrated Omics Approach with Epigenomic Focus
Authors: Chakraborty Nabarun , Muhie Seid , Gautam Aarti , Hammamieh Rasha , Jett Marti 
Published: November 2023

Our previous transcriptomic study [1] suggested potential positive significance of the effects of fish oil on the cerebralneurogenesis and neuroprotection, and contrasting therapeutic implications for the neurodegenerative disorders. Stimulating the calcium‐induced growth cascade and downstream PI3K‐AKT‐PKC network, fish oil augmented neuritogenesis, reinforced synapse and potentially promoted long‐term potentiation. In conjunction, this diet reduced the amyloidal burden, attenuated oxidativestress, and assisted in somatostatin activation. Therefore, we anticipated the potential therapeutic roles of fish oil in attenuating Alzheimer's disease, Parkinson's disease, and affective disorder. Motivated from this past study, we investigated the shift of the DNA methylation and microRNA profiles caused by these same dietary supplements and potential consequences on cerebralfunctionalities.N‐3 PUFA‐enriched diet (ERD, n3:n6= 7:1) and balanced diet (BLD, n3:n6= 1:1) were customized to contain caloric content equivalent to the standard lab diet (STD, n3:n6= 1:6). The diets were administered to three groups of C57BL/6J male mice from their weaning age until late adolescence. At the study endpoint, the transcriptomic and epigenomic profiles of the hemi brains were probed.Our primary focus was on hypermethylated probes and microRNAs which were causally related to transcriptional silencing. We found 18 overexpressed microRNAs in the ERD‐fed mice that inhibited the transcriptional expressions of 580 genes.. Likewise, there were 40 overexpressed microRNAs targeting 800 suppressed gene in BLD‐fed mice; a majority of these microRNAs are functionally associated with the neurodegenerative disorders, such as schizophrenia and Alzheimer's disease.In addition, there were ~4,000 probes hypermethylated in either ERD‐ or BLD‐fed mice. Of particular interest, ERD‐fed mice exhibited ~300 genes concurrently imprinting hypermethylated CpG islands and silenced transcripts. Similarly, 131 genes displayed hypermethylated CpG islands and transcriptional suppression in BLD‐fed mice. All these hypermethylated CpG islands were mapped within 50kb from their respective transcriptional startsites, indicating their rather archetypal effects on transcriptional silencing. Functional annotation of these genes indicated a pattern of neurological implications distinguishing the positive effects related to ERD and BLD. ERD primarily influenced the developmental phase of neuronal and cerebral activation, while the behavioral manifestations were potentially stimulated by BLD. A validation study is underway.Disclaimer: Research was conducted in compliance with the Animal Welfare Act, and all other Federal requirements. The views expressed are those of the authors and do not constitute endorsement by the U.S. Army.
The role of 12/15‐Lipoxygenase in a Mouse Model of Alcoholic Liver Disease
Authors: Kirpich Irina , Miller Matthew , Ramsden Christopher , Feldstein Ariel , McClain Craig 
Published: November 2023

Introduction/Aim12/15‐Lipoxygenase (12/15‐LOX) catalyzes the oxidation of free and esterified polyunsaturated fatty acids, predominantly, linoleic and arachidonic, generating the whole spectrum of bioactive lipid mediators. This enzyme is involved in the regulation of various homeostatic processes as well as in the pathogenesis of multiple diseases. Recently published studies from our group and others have shown that experimental and clinical alcoholic liver disease was associated with elevated oxidized linoleic acid metabolites (OXLAMs), specifically 9‐ and 13‐hydroxy‐octadecadienoic acids (9‐ and 13‐HODEs). The aim of the present study was to examine the role of 12/15‐LOX in an animal model of ALD.Materials and MethodsEight‐week‐old male 12/15‐LOX knockout mice (12/15‐LOX−/−), and their genetically unaltered wild type (WT) counterparts were fed Lieber–DeCarli control (isocaloric maltose‐dextrin) or ethanol (5% w/v) liquid diets ad libitum for 10 days plus a single binge ethanol administration (5 g/kg) by gavage, whereas mice in control groups were gavaged with isocaloric maltose dextrin. Liver histopathological changes were estimated by hematoxylin and eosin staining. Liver triglyceride accumulation was assessed by biochemical assay and Oil Red O staining. Liver injury was evaluated by plasma ALT and AST activity. Liver inflammation was determined by hepatic proinflammatory gene expression. Chloroacetate esterase staining was used to evaluate necroinflammatory changes. TUNEL staining was used to determine hepatic apoptotic cell death. Plasma OXLAMs were analyzed on LC‐MS and GCxGC/TOF‐MS platforms.ResultsChronic‐binge alcohol administration resulted in a marked increase in plasma OXLAM levels, specifically 9‐HODE and 13‐HODE, in parallel with up‐regulation of hepatic 12/15‐LOX in WT animals. These effects were associated with hepatic steatosis, inflammation and injury. 12/15‐LOX genetic depletion did not reduced hepatic steatosis caused by EtOH, but did ameliorate hepatic injury as assessed by ALT (231.2+32.8 U/L in WT vs 148.4+48.6 U/L in 12/15‐LOX−/−) and AST (279.1+36.7 U/L in WT vs 177.1+28.3 U/L in 12/15‐LO−/−) levels. 12/15‐LOX deficiency did not affect hepatic apoptosis; while neutrophil infiltration was significantly reduced in 12/15‐LOX deficient mice compared to WT animals. 12/15‐LOX deficiency prevented EtOH‐induced pro‐inflammatory cytokine and chemokine expression, including Tnf‐α and Mcp1 mRNA levels.Conclusion12/15 LOX plays an important role in the pathogenesis of experimental ALD, in part, via increased production of OXLAMs. 12/15‐LOX and OXLAMs may represent novel therapeutic targets in ALD managementSupport or Funding InformationStudy supported by the NIH and Veterans Administration.
Effects of Synthetic Ligands on Heterodimer Pairs Regarding PPARα, RXRα and LXRα
Authors: Delman Emily Erin , Rider Stanley Dean , Hostetler Heather Ann 
Published: November 2023

In this study, we hypothesize that ligand binding determines the binding partners and subcellular localization of nuclear receptors involved in atherosclerosis and diabetes. Specifically, we are interested in the effects of synthetic ligands on the peroxisome proliferator‐activated receptor alpha (PPARα), the liver X receptor alpha (LXRα) and the retinoid X receptor alpha (RXRα) and corresponding effects on heterodimer formation. Binding assays using a photon counting spectrofluorometer indicate promiscuity of protein‐ligand interactions where ligand binding is not solely exclusive. Additionally, Forster Resonance Energy Transfer (FRET) assays indicate that synthetic ligands affect the protein binding capability of the novel heterodimer PPARα:LXRα. Changes in secondary structure are anticipated, which would argue certain synthetic compounds can induce conformational changes that alter heterodimer formation. Bimolecular Fluorescence Complementation (BiFC) allows us to view protein‐protein interactions in vivo using fluorescence microscopy. We show that synthetic ligands influence subcellular localization of these nuclear receptors in stably transfected living cells. Due to the complex regulation of these nuclear receptors, it becomes necessary to study synthetic ligands with each receptor, individually and in heterodimeric form. Our findings will contribute to further the understanding of the clinical implications of synthetic ligands on disease states such as atherosclerosis and diabetes.Support or Funding InformationThis research is supported by USPHS NIH grant DK77573 and the Emerging Science Seed Grant provided by the Boonshoft School of Medicine and Wright State University.
Critical Role of Residues in Helices 9 and 10 of the Liver X Receptor Alpha (LXRα) in Mediating Heterodimer Choice between Heterodimeric Complexes
Authors: Bedi Shimpi , Hostetler Heather A. , Rider Stanley Dean 
Published: November 2023

The Liver X Receptor Alpha (LXRα) is a ligand‐activated nuclear transcription factor that is activated upon binding to oxysterols and plays an important role in lipid homeostasis. Heterodimerization with a common heterodimeric partner, the retinoid X receptor (RXR), regulates the transcription of target genes. LXRα‐RXRα activation has been shown to inhibit cardiovascular disease and reduce atherosclerotic lesions in mice. Additionally, LXRα can also form heterodimers with the Peroxisome Proliferator‐Activated Receptor Alpha (PPARα), another ligand‐activated nuclear receptor involved in fatty acid (FA) metabolism. Key residues located in helices 9 and 10 of LXRα may mediate dimerization of LXRα with partner receptors. These residues show a high degree of conservation across species and among nutrient‐sensing nuclear receptors. Little is known about the effects of charge disruption at the LXRα interface on protein‐protein and protein‐ligand associations. Using overlap PCR, we generated LXRα mutants in which the interface had been modified to determine whether heterodimerization selective variants of LXRα can be generated. Fluorescence spectroscopy in cells and in vitro demonstrated that charge reversal at the LXRα dimerization interface has the ability to influence LXRα protein and ligand binding. Using the intrinsic quenching of the aromatic amino acids located in the ligand binding pocket of LXRα, we have demonstrated that LXRα R415A does not bind T‐0901317, but retains its ability to bind 25‐hydroxycholesterol and palmitic acid. Direct protein‐protein binding experiments using the fluorescently labeled proteins suggested that LXRα R415A binding to RXRα is unaffected whereas binding to PPARα is enhanced. Another mutant of LXRα leaves LXRα‐PPARα binding intact but reduces LXRα‐RXRα binding. This indicates that key residues at the interface differentially regulate protein‐protein and protein‐ligand interactions of LXRα. Key contact regions of LXRα may represent a new drug target for modulating LXRα signaling in lipid‐related disorders.Support or Funding InformationThis research was supported by USPHS NIH grant DK77573 and the Emerging Science Seed Grant provided by the Boonshoft School of Medicine and Wright State University.
Characterization of the Functional Significance of PPAR/LXR Heterodimers via a Novel Response Element
Authors: Klingler Andrea M , Rider S. Dean , Hostetler Heather 
Published: November 2023

Peroxisome proliferator‐activated receptor alpha (PPARα) and liver X receptor alpha (LXRα) are nuclear receptors known to regulate opposing processes in energy homeostasis through heterodimerization with the retinoid X receptor alpha (RXRα). Dysregulation of these proteins has been linked to such metabolic disorders as atherosclerosis, hypercholesterolemia, and hypertriglyceridemia. Among the more intriguing recent discoveries in the field is that PPARα and LXRα are also able to interact directly with each other and bind DNA as a heterodimer. Although this interaction is widely thought to be inhibitory or otherwise non‐functional, we hypothesize that PPARα‐LXRα heterodimers are indeed capable of carrying out regulatory function via previously uncharacterized response elements. Gel shift analysis of several designed direct repeat sequences revealed that PPARα‐LXRα heterodimers bind preferentially to a DR4 motif, and supershift assays confirmed both PPARα and LXRα were bound to the oligonucleotide probes. Similarly, intrinsic fluorescence quenching assays determined that PPARα‐LXRα binds to these DR4 sequences with high affinity. The sequence bound with the highest affinity was found to occur naturally in the human genome, including near the promoter regions of known PPARα and/or LXRα target genes, such as apolipoprotein A‐1 (APOA1), cytosolic sulfotransferase 2A1 (SULT2A1), and the sterol regulatory element binding protein 1‐c (SREBP‐1c). Dual luciferase reporter assays showed that overexpressing PPARα and LXRα in COS‐7 cells leads to transactivation of the endogenous APOA1 and SULT2A1 genes, but not of SREBP‐1c. These findings illuminate the complex regulatory mechanisms modulated by PPARα and LXRα, and provide a potential new avenue for targeting these receptors for treatment of metabolic disorders.Support or Funding InformationThis work was supported by USPHS NIH grant DK77573 and the Emerging Science Seed Grant provided by the Boonshoft School of Medicine and Wright State University.
Coiled‐coil domain containing protein 103 regulates myeloid development and function
Authors: Beckman Sarah , Waxman Josh 
Published: November 2023

RationaleAbnormal myeloid function is implicated in many diseases, including leukemia, atherosclerosis and others. Therefore, understanding regulators of myelopoiesis and myeloid function will lead to novel therapies. We have identified that the poorly understood coiled‐coil domain containing protein 103 (Ccdc103) is expressed in human and zebrafish myeloid cells.ObjectiveThe goal of this project is to test the hypothesis that Ccdc103 is a novel regulator of myeloid development and function.Methods and ResultsCcdc103 is known to be involved in cilia motility and dynein arm assembly through direct interactions with microtubules. However, we found previously unrecognized expression of Ccdc103 in zebrafish and human myeloid progenitor cells. Furthermore, CCDC103 is expressed in highly proliferative myeloid leukemia cell lines and proliferative cultured myeloid progenitors, suggesting Ccdc103 may promote proliferation. Consistent with the hypothesis that Ccdc103 is involved in myeloid production, there are decreased and increased neutrophils and macrophages, respectively, in ccdc103 mutant and ccdc103 overexpressed zebrafish embryos. In addition to the defects in myeloid cell number, we found that Ccdc103 deficient embryos display abnormal myeloid progenitor cell migration, with myeloid progenitor cells migrating inappropriately from the anterior blood island. To further test the idea that Ccdc103 plays a role in myeloid migration, we performed in vivo wounding assays in zebrafish embryos at 24 hours post fertilization. We find that Ccdc103 deficient neutrophils and macrophages have a reduced ability to directly migrate to the wound and have abnormal macrophage morphology.Conclusion(sAltogether, our results indicate that Ccdc103 has functions outside of cells with motile cilia as a novel regulator of myeloid production and directed migration, indicating it or processes it regulates may be exciting targets for confronting myeloid‐related cardiovascular disorders.Support or Funding InformationR01 HL112893 to JSW. NIH T32 Understanding of Cardiovascular Disease Mechanisms to SAB.