bluuebunny/crossref_metadata_2025_split huggingface dataset:
Thimet Oligopeptidase (EP24.15), a Neuropeptide Processing Enzyme Regulating Xenin Signaling
Authors: Glucksman Marc , Philibert Keith , Mizuno Tooru , Lew Piesan 
Published: November 2023

Xenin is a 25 amino acid neuropeptide with sequence homology and actions similar to hypothalamic and ileal neurotensin (a satiety factor) as well as binding to the neurotensin receptor. Xenin is produced by a subgroup of chromogranin A+ cells in the duodenum and this neurohormone is involved in glucose homeostasis and increases the response to glucose‐dependent insulinotropic peptide (GIP). Upon its release after ingestion Xenin inhibits secretion of pancreatic exocrine peptides in the gastrointestinal tract. The mode of regulating the peptide hormone Xenin remains elusive. The metalloendopeptidase EC 3.4.24.15 (thimet oligopeptidase, EP24.15) has been demonstrated to play a key role in the cleavage and subsequent regulation of several neuropeptides that also exist in the gut such as neurotensin and somatostatin. A systems biology approach to regulating signaling beginning from high throughput structural analyses, in silico molecular modeling and potential substrate screening implicated Xenin as a substrate of EP24.15 and thus, this enzyme as a possible regulator of Xenin signaling.To substantiate a potential regulatory mechanism for the functioning of Xenin, we first identified potential active site binding and cleavage sites, measured enzymatic parameters, performed structural studies, and determined if EP24.15 and Xenin are co‐expressed in gut regions relevant to glucose homeostasis. After proprietary in silico modeling paradigms identifying a potential favorable interaction, EP24.15 and Xenin were co‐incubated and subjected to matrix‐assisted laser desorption ionization time‐of‐flight (MADLI‐TOF) mass spectrometry to confirm that EP24.15 can cleave Xenin in vitro. Enzyme kinetics were performed with standards via high performance liquid chromatography yielding results consistent with known substrates. Notably, Xenin is cleaved in a homologous manner as neurotensin. Double‐label immunohistochemistry demonstrated Xenin and EP24.15 immunoreactivity within the mouse small intestine and specifically co‐expression within the intestinal mucosa and submucosa. Furthermore, there is co‐expression of EP24.15 and Xenin in cells of both human and mouse stomach in the gastric mucosa. Taken together this data suggests that EP24.15 may act to cleave Xenin in vivo and represents an additional facet of the control mechanism of glucose homeostasis. As a potential pharmacological therapeutic target, understanding regulation of Xenin by the neuropeptide processing enzyme, EP24.15, may provide insight into an alternative strategy for glucose regulation and diseases such as diabetes and obesity.Support or Funding InformationSupported by NIH OD010662 (MJG) and CIHR MOP123208 (TMM)
Effect of Surfactin Administration on Glucose and Lipid Metabolism in Mice Fed a High Fat Diet
Authors: Santos Maria Emilia Soares Martins , Porto Marcos Augusto Nascimento , Marinho Pablo Rayner Sousa , Oliveira Michael Eder , Granjeiro Paulo Afonso 
Published: November 2023

The high intake of high fat diet has been related with several changes on glucose and lipid metabolism and also on gut microbiota composition. Surfactin is an active compound produced by a wide variety of bacteria, fungi, and yeast that have important biological activities as anti‐tumour, anti‐microbial and anti‐inflammatory. Oral probiotics or their secreted factors are becoming the novel strategy for treatment of metabolism alterations. The aim of this study was to determine the influence of surfactin produced by Bacillus subtilis (ATCC 19659) on glucose and lipid metabolism in mice fed a high fat diet. Male mice (n=6/group) were fed with a control (C) or high fat (HF) diets and water ad libitum for 10 weeks. One group (HFS) was orally administrated with 5 mg/kg body weight of Surfactin once a week for 4 doses. Surfactin was isolated and purified from Bacillus subtilis culture. It was lyophilized and later dissolved in phosphate buffered saline (PBS) before the administration. Food (g) and water intake (ml) were measured daily. Body weight (g) was evaluated weekly. After 10 weeks, mice were euthanized and blood was collected. Using commercial kits the levels (mg/dl) of glucose (GLU), triglycerides (TG), total cholesterol (CHO), HDL and VLDL cholesterol were measured. Epididymal (EAT) and retroperitoneal (RAT) adipose tissue depots were collected and weighted (g). Results were expressed as mean ± SEM. Data were analyzed using T‐test. P<0.05 was considered significant for all data.No difference was observed in body weight between the groups however HF and HFS groups eat and drink less than C. An increase of ~47% at EAT was observed in HL (2.69±0.31) vs C (1.83±0.18). There was no difference at RAT in any group. There was an increase of ~27% in HF (362.7±11) vs C (284.9±30) and a decrease of ~20% in HFS (289.3±34) vs HF (362.9±11) at GLU. An increase of ~47% at CHO in HF (155.5±10.9) vs C (105.6±3.4) and also in HDL (~59%) HF (112.4±6.5) vs C (70.4±3.9) was observed. TAG was decreased ~32% in HF (109.8±2.9) vs C (162.9±19.1) and also in HFS (~30%) (76.4±11) vs HL (109.8±2.9). Same pattern of results were observed in VLDL.In conclusion, we observed that HF diet intake was able to disrupt glucose and lipid metabolism and surfactin seems to be efficient in the control of glycemia and tryglyceredemia. We hipothesize that these effects achieved are due surfactin action on anti‐inflammatory cytokines production and/or changes in gut microbiota composition.Support or Funding InformationCNPq
Relation of Hypoglycemic Activity and the Antioxidant Capacity of Justicia Spicigera Leaf Extracts in Diabetic Rats
Authors: Martínez‐Mora José A. , Murillo‐Villicaña Marina , Salgado‐Garciglia Rafael , Manzo‐Avalos Salvador , Cortés‐Rojo Christian , Montoya‐Pérez Rocío , Saavedra‐Molina Alfredo 
Published: November 2023

Diabetes mellitus is a metabolic disorder characterized by chronic hyperglycemia with disturbances in the metabolism of carbohydrates, lipids, and proteins, where there is an absolute or relative deficiency in the secretion and/or insulin action, or both, reducing the quality and life expectancy of those who suffer, as it promotes the generation of various diseases such as renal disease and neuropathies, among others; hyperglycemia being the cause of complications of diabetes by a mechanism involving oxidative stress. Since ancient times, diabetics have used medicinal plants to maintain normal blood sugar levels. Justicia spicigera (J. spicigera) is a branched and medicinal erect shrub that is used to treat diabetes. The objetive of the project is: To analyze the effect of extracts from the leaves of J. spicigera on levels of glucose, urea, creatinine, cholesterol, triglycerides and activity of mitochondrial antioxidant enzymes in diabetic rats. The plant J. spicigera was used to prepare 3 extracts with solvents of different polarity (hexane, ethyl acetate and ethanol) at 100 mg/kg, and administered for 30 days on a daily basis (1ml) orally to diabetic rats streptozotocin induced. Therefore eight groups of rats were assigned: 1) Normoglycemic control, 2) Diabetic control, 3) Normoglycemic + hexane extract, 4) Diabetic + hexane extract, 5) Normoglycemic + extract ethyl acetate, 6) Diabetic + acetate ethyl, 7) Normoglycemic + ethanol extract, 8) Diabetic + ethanol extract. Our study revealed that the administration of ethyl acetate and ethanol extracts of leaves of J. spicigera have hypoglycemic activity in diabetic rats. The intake of ethanol extract increases levels of urea in normoglycemic rats and increases triglyceride levels in diabetic rats. Meanwhile, administration of the hexane extracts, ethyl acetate and ethanolic did not modify urea levels in diabetic rats, however, they did increase creatinine levels of normoglycemic rats. Hexane and ethyl acetate extracts decrease levels of cholesterol and triglycerides in diabetic rats. It can be concluded that, because each solvent drag different types of compounds expected the different effects obtained in normoglycemic and diabetic rats.Support or Funding InformationAcknowledgements: The authors appreciate the partial economic support by CIC‐UMSNH (2.10 to RSG; 2.16 to ASM; 2.37 to SMA).
Development of Novel Allele Specific PCR Based Assays to Investigate the Contribution of Cortisol to Metabolic Syndrome
Authors: Godlewski Brianna R. , Salm Justin R. , Cohen Brian D 
Published: November 2023

A diagnosis of Metabolic Syndrome (MSX) requires patients to present with three or more of the following symptoms: elevated fasting blood glucose levels, elevated serum triglyceride levels, low serum HDL levels, elevated blood pressure, and truncal obesity. This pathology shares many similarities with Cushing's Syndrome (CS) but a diagnosis of CS requires hypercortisolemia. This similarity has led our lab and others to hypothesize that MSX may be a Cushingoid‐like state caused by hypersensitivity of the glucocorticoid receptor (GR), a nuclear hormone receptor that is activated when cortisol binds. Using an allele specific polymerase chain reaction (PCR) protocol, previous research in our lab has found significant correlations between the BclI and N363S polymorphisms of the glucocorticoid receptor and altered patient metabolic profiles in a population of patients seeking bariatric surgery, suggesting their possible contribution to diseases such as obesity and Metabolic Syndrome. Recent work in our lab has expanded our study to include the cortisol resistance polymorphism TthIIIi (rs10052957) to test the hypothesis that there would be a decreased prevalence of this polymorphism in our subject population. The TthIIIi allele is present at a lower frequency in our research population compared to reported frequencies, consistent with our hypothesis.In addition to investigating the glucocorticoid receptor, we have also developed an allele specific PCR assay for previously identified single nucleotide polymorphisms (rs12086634 and rs846910) in the cortisol reductase enzyme, also known as 11beta‐hydroxysteroid dehydrogenase (11β‐HSD) type 1. The 11β‐HSD1 enzyme is responsible for the conversion of inactive cortisone to cortisol, and works primarily in the adipose tissue. The two polymorphisms listed above are both associated with overactivity of the enzyme and are another possible cause for the Cushingoid‐state observed in MSX patients. In our patient population, the frequency of heterozygosity for the rs12086634 SNP was nearly 50% higher than the reported frequency in the general population, suggesting that this polymorphism may contribute to increased obesity and the MSX metabolic profile by increasing the localized cortisol in adipose tissue. Greater understanding of the interplay between these single nucleotide polymorphisms can help physicians and patients make more informed decisions about treatment options for obesity and metabolic syndrome.
FoxO1 localization changes in Cinnamonium cassia treated 3T3‐L1 pre‐adipocytes
Authors: Bova Kaitlin , Aulthouse Amy , Stockert Amy 
Published: November 2023

With increased focus on diabetes treatment and prevention, characterizing the mechanisms of action of various alternative treatments becomes increasingly important. Numerous studies have shown that cinnamon from a variety of species lowers both post‐prandial and fasting blood glucose levels. Our research is focused on examining the effects of aqueous extract of Cinnamonium cassia on a variety of signaling mechanisms normally affected by insulin. One such mechanism utilized by insulin is the Akt pathway which leads ultimately to the phosphorylation of FoxO1, a transcription factor involved heavily in metabolism. Nucleation of the FoxO1 factor is dependent on its phosphorylation state. This study is aimed at determining the effects of cinnamon treatment on the FoxO1 localization. Preliminary studies indicate minimal effects on expression level of FoxO1, indicating that if involved, FoxO1 localization is likely the key to lowering blood glucose levels. We completed a multi‐time point study looking at early and later differentiation stages if the 3T3‐L1 cells both in the presence and absence of cinnamon treatment. Cells were divided into two groups, those which were chemically induced for differentiation and those which received no chemical induction. RNA samples were collected from each time point and treatment groups for synthesis of cDNA followed by quantitative PCR. Although changes in expression level were observed, large changes in protein level were not observed and it is unlikely that expression level of FoxO1 plays a role in the observed clinical outcomes. In order to explore the effects of treatment on localization of FoxO1, protein extractions were conducted isolating both the nuclear and the cytosolic proteins. These samples were analyzed by Western blot to identify what effect cinnamon treatment had on nucleation of the FoxO1. Preliminary analysis of these data suggest that cinnamon facilitates nuclear exclusion.Support or Funding InformationOhio Northern University Academic Affairs Summer Research Support Program
Analysis of Gene Expression in a Female Rat Model of Diet‐induced Non‐alcoholic Fatty Liver Disease Using RNA‐sequencing
Authors: Bowman John James , Lee Jennifer Karin , Perets Ron A , Knabe Melina L , Guider Jamie W , Phillips Scott , Sarwadnya Uma D , Blythe Sarah N , Toporikova Natalia , Whitworth Gregg B 
Published: November 2023

We have established a diet‐induced model of non‐alcoholic fatty liver disease (NAFLD) in the adult female rat, which we have characterized through RNA‐seq, histology, and physiological measurements. Furthermore, we investigated whether peripheral estrogen signaling has affected this disease. All rats were ovariectomized, with basal levels of estrogen restored in half the animals by implantation of a slow release estradiol pellet. We observed a significant effect of both diet and estrogen on animal weight gain, with animals fed a high‐fat, high‐sugar diet without supplemental estrogen demonstrating a >2‐fold increase in weight gain compared to control diet animals that received estradiol pellets. Our high‐fat, high‐sugar animals demonstrate physiological and histological markers of NAFLD, including steatosis, inflammation, and an increase in connective tissue in the liver. Additionally, we measured percent body fat, liver weight, percent liver fat, and insulin, glucose, triglycerides, and cholesterol levels. Moreover, we used RNA purified from liver tissues to quantify global changes in gene expression between our experimental groups through RNA‐seq, with the goal of identifying genes that are differentially expressed in the NAFLD induced animals. In addition, the quantification of certain promising gene targets using RT‐qPCR serves as validation for RNA‐seq. Interestingly we see the greatest effect on gene expression through the interaction between diet and estrogen. We will be discussing the in‐depth, high resolution gene analysis done on the liver tissues as it relates to the induction of this disease. A few initial pathways and processes that are upregulated in our high‐fat animals include the inflammatory cytokines and genes involved in lipid metabolism.Support or Funding InformationHoward Hughes Medical InstituteJeffress Trust Awards Program Interdisciplinary ResearchR.E. Lee Summer Research Scholars
Micro RNAs Mediating Effects of Adipose Angiotensinogen in Adipocyte Inflammation and ER Stress
Authors: Menikdiwela Kalhara Rashmikumara , Moustaid‐Moussa Naima , Ramalingam Latha , Scoggin Shane , Kalupahana Nishan Sudheera 
Published: November 2023

Obesity is a complex disease and a global epidemic associated with chronic low grade inflammation and metabolic dysregulation. The renin‐angiotensin system (RAS) plays a vital role in controlling blood pressure and fluid balance. Although RAS components are generally overexpressed in adipose tissue in obesity, mechanisms by which RAS induces adipose endoplasmic reticulum (ER) stress and inflammation, both hallmarks of obesity, are not well understood. Here, we hypothesized that overproduction of angiotensinogen (Agt, precursor protein in RAS), induces ER stress and inflammation in adipocytes, thereby, contributing to obesity‐associated metabolic alterations. To test this hypothesis, we used the aP2 promoter‐driven Agt overexpression in adipose tissue of transgenic (Agt‐Tg) mice, compared to wild type mice (Wt), with or without a RAS inhibition (captopril, ACE inhibitor). We also used 3T3‐L1 adipocytes to test direct effects of angiotensin II and RAS inhibitors in obesity‐associated inflammation and ER stress.The serum levels of pro‐inflammatory markers such as MCP‐1 (Monocyte Chemoattractant Protein‐1) were significantly higher in Agt‐Tg compared to Wt mice. By contrast, anti‐inflammatory cytokines such as IL (interleukin)‐10 and IL‐2 were significantly lower in Agt‐Tg mice compared to Wt mice. Further, mRNA levels for ER stress markers, including BiP, ATF4 and XBP1 and cell apoptosis marker, CHOP, were significantly higher in Agt‐Tg mice compared to Wt mice. To gain further insight into mechanisms and pathways mediating Agt effects on ER stress and RAS‐associated obesity and inflammation, we conducted miRNA profiling by RNA Seq in adipose tissue from Agt‐Wt and Wt mice. We identified 3 miRNAs that were upregulated and 14 miRNAs that were downregulated in adipose tissue from Agt‐Tg vs. Wt mice. These were further validated by real time quantitative PCR, and identified miRNA 708 and 143 to be upregulated, while miRNA 30 family was down regulated in Agt‐Tg mice compared to Wt mice. These results are consistent with previously reported roles for these miRNAs in ER stress. Moreover, Agt‐Tg mice treated with Captopril had a significantly lower expression of CHOP and ATF4 mRNAs compared to untreated controls. In agreement with these findings, captopril prevented angiotensin‐II (Ang II) induced expression of ER stress genes in 3T3‐L1 adipocytes. Further studies are ongoing in cultured adipocytes to dissect molecular mechanisms linking our candidate miRNAs to RAS, ER stress and inflammation.In conclusion, overexpression of Agt in adipose tissue increased inflammation and ER stress. These effects are possibly mediated in part by miRNAs 708, 143 and 30 and by direct Ang II actions in adipocytes. These miRNA‐regulated pathways could be used as potential therapeutic targets to reduce obesity‐linked inflammation and ER stress.
Time Resolved IRMS Study of Staurosporine Induced Apoptosis in Murine Astrocyte Cells
Authors: Weber Kayla N , Husher Sara C , Mazzer Paula A 
Published: November 2023

The advent of genomic and post genomic technologies is transforming the field of toxicology, and is moving the discipline from the realm of classical biology into those of molecular biology and bioanalytical chemistry. At the molecular level, it is possible to focus on both the direct impact of toxic insult as well as the cellular or organismic response to the resulting damage. The “‐omics” technologies—genomics, proteomics, and metabolomics—are likely to lead the way in the development of approaches to molecular pathology. Metabolomics, in fact, shows such promise in understanding the biochemical functioning of cells that the development of metabolomic technologies has been a major NIH roadmap initiative. One subspecialty of metabolomics, lipidomics, shows a special promise because lipids are simple to analyze and have short response time to toxic insult. One shortcoming of current lipidomic exploration is a lack of knowledge of which lipid synthetic pathways are activated in response to toxic insult. Therefore, we undertook to study lipid synthesis in response to apoptotic cell death in murine (C8‐D1A ATCC) astrocyte cells.To investigate time resolved lipid expression we used a model of staurosporine‐induced apoptosis in murine astrocyte cells. We extracted lipids at defined time points after staurosporine insult using a modified Bligh and Dyer technique. Lipids were then sorted by lipid class using rapid HPTLC and then derivatized to fatty acid methyl esters (FAMEs). Time dependent changes in fatty acid expression were observed over the first six hours following staurosporine treatment. Inclusion of 13C‐labeled glucose immediately prior to staurosporine addition allowed for investigation of de novo fatty acid synthesis by Isotope Ratio Mass Spectroscopy (IRMS). The ability to use IRMS to trace formation of lipid synthesis has the potential to add a new dimension to future lipidomic studies.Support or Funding InformationThis work was sponsored by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103443.
Readers, writers and erasers of nuclear PIP3.
Authors: Blind Raymond D 
Published: November 2023

Structural analyses of the NR5A nuclear receptors (NR5A1 and NR5A2) show that PIP2 and PIP3 bind these transcription factors in a format where the acyl chains are buried deep in the core of the NR5A protein, while the phospholipid headgroups are solvent exposed. We previously demonstrated that the phosphoinositide headgroups of these complexes are accessible to nuclear lipid signaling enzymes, which directly modify these transcription factor‐bound PIPs, altering the transcriptional output from NR5As. Here, we took a proteomics approach to ask if the solvent exposed PIP3 headgroup is available to interact with nuclear proteins. We identified two PH‐domain containing proteins that interact with NR5A1/PIP3, but not other phospholipid bound forms of NR5A1, and siRNA knockdown of these two genes decreases NR5A1 transcriptional activity. PH‐domains are one of the most well represented domain classes in the human genome, and are widely thought to mediate membrane binding, however only 10% of the PH‐domain proteins in yeast bind membranes. Our data suggest a fraction of PH‐domain containing proteins act as readers of nuclear PIP3, while nuclear lipid signaling enzymes acts as writers and erasers of nuclear PIP3. These data provide a direct mechanism that explains how nuclear PIP3 mediates transcriptional activation in mammalian cells.Support or Funding InformationJimmy V Foundation V‐Scholar Young Investigator Award, Vanderbilt Diabetes Research and Training Center Pilot & Feasibility Award on NIDDK P30 DK020593, Vanderbilt Diabetes Center Discovery Award, American Cancer Society/Vanderbilt‐Ingram Cancer Center Institutional Research Grant IRG‐58‐009‐56.