bluuebunny/crossref_metadata_2025_split huggingface dataset:
Potent lipidated antagonists to protease‐activated receptor 2 (PAR2)
Authors: Boitano Scott , Hoffman Justin , Flynn Andrea , Sherwood Cara L. , Zhang Zhenyu , Patek Renata , Price Theodore J. , Vagner Josef 
Published: November 2023

Protease‐activated receptor‐2 (PAR2) belongs to a four‐member family of G‐Protein coupled receptors (GPCRs) that contain internal ligands exposed following exogenous or endogenous protease cleavage of the extracellular amino terminus. PAR2 is associated with a variety of inflammatory conditions, including asthma and pain. The contributions of PAR2 signaling to disease has been hindered by the lack of potent, efficacious antagonists, and their potential for biased‐ligand signaling. We recently demonstrated that lipid tethering of known PAR2 peptidomimetic agonists based on the primary trypsin cleavage sequence (SLIGRL) increased their potency > 200 fold. In this study, we used lipid tethering (hexadecyl (Hdc) group with polyethylene glycol (PEG) spacers) and heterocycle (2‐aminothiazoyl; 2‐at) substitution of hexapeptide sequence derived from the primary cleavage site of kallikreins 4/16 (SSKGRS) to elucidate novel PAR2 antagonists. Compound 562 (C562), 2‐aminothiazol‐4yl‐SKGRS‐PEG3‐Hdc blocks PAR2 Ca2+ signaling elicited via peptidomimetics (2‐at‐LIGRL‐NH2) or via asthma associated protease activation (Alternaria alternata filtrates) in cultured human bronchial epithelial cells (16HBE14o‐). This compound was a biased‐signaling antagonist in that it had no effect on mitogen activated protein kinase (MAPK) signaling, the other major signaling pathway activated via PAR2. A shortened version of C562, 2‐at‐SKGR‐PEG3‐Hdc (C595), maintained antagonistic activity against peptidomimetic activation in an in vitro physiological signaling assay (xCELLigence). C595 is closely related to the previously described potent and specific PAR2 agonist, 2‐at‐LIGR‐PEG3‐Hdc. Thus, we screened a series of potential PAR2 ligands with a heterocycle serine substitute followed by four amino acids (XXGR) and the PEG3‐Hdc lipid tether. We describe several potent agonists, and one partial agonist (C608, 2‐at‐TIGR‐PEG3‐Hdc) that also acts as a potent, specific and biased signaling antagonist of PAR2. When used in nanomolar concentrations, C608 blocked PAR2‐dependent Ca2+ signaling via protease or peptidomimetics without effects on MAPK signaling. C562, C595 and C608 are novel pharmacological tools that can be used to evaluate the physiological consequences of PAR2 full and biased ligand signaling.Support or Funding InformationThis work was primarily funded by a multi‐PI grant (NS 073664 to SB, JV and TJP) from the National Institutes of Health. Additional support for this work was from the following grants: National Institute of Health training grants (T32 HL 007249 for ANF; T32 ES 007091 for CLS), National Institutes of Health R01NS 065926 (TJP) and R01AI083403 (SB; Michael O. Daines, PI), State of Arizona Technology and Research Initiative Fund Awarded through Bio5 (JV).
Effect of frequent intake of sweeteners on the JAK2/STAT3 signaling pathway in the central nervous system of mice
Authors: Barrios Alberto Andrés , Contreras Irazú , Estrada José Antonio 
Published: November 2023

Sweeteners are food additives that are commonly used worldwide. There are discrepancies in epidemiological studies evaluating their effect on body weight and appetite; this generates the need to further study the mechanism by which sweeteners could affect satiety and energy balance.This study's main objective is to evaluate whether frequent intake of the commercial sweeteners; sucralose, steviol glycosides and sucrose could cause alterations in the leptin‐mediated JAK2/STAT3 signaling pathway, in the central nervous system of mice.For this purpose, three study groups comprised of 14‐week‐old female BALB/c mice were established: sucrose, steviol glycosides and sucralose groups were supplemented with those sweeteners in their daily water for 6 weeks (from week 8 to week 14), additionally, a control group was established without sweetener supplementation. Mice were fed ad libitum and weight was measured weekly. The amount of food and water intake was measured daily. After 6 weeks, body composition of mice was determined using bioelectrical impedance and total brain proteins were obtained to determine the expression of total and phosphorylated JAK2 and STAT3 proteins by western blot.Our preliminary results show that there is a higher consumption of water, but lower food intake in the sucrose group compared with the control and other experimental groups; there are no significant differences in body weight and adiposity among groups. Regarding JAK2/STAT3, expression of pJAK2 and JAK2 was found to be higher in the group supplemented with sucralose, whereas we did not observe changes in the STAT3 proteins. We also found that mice supplemented with steviol glycosides and sucrose had a lower expression of pJAK2 and JAK2 compared to control mice.Our results suggest that frequent intake of commercial sweeteners may cause alterations in energy balance and appetite by altering the phosphorylation of JAK2, however, we need further study the effect of these sweeteners in the leptin‐receptor expression and serum leptin concentrations.
Mitochondrial Translocator Protein (TSPO) Is a Regulator of Fatty Acid Metabolism
Authors: Tu Lan N , Zhao Amy H , Hussein Mahmoud , Selvaraj Vimal 
Published: November 2023

Translocator protein (TSPO), first identified as a secondary target of benzodiazepines and termed the ‘peripheral’ benzodiazepine receptor (PBR), is a highly conserved protein in the outer mitochondrial membrane. Despite intensive research for the past 25 years, the exact function of TSPO remains elusive. High TSPO expression seen in steroidogenic mitochondria, and induction of steroid hormone production by TSPO binding drugs, led to a proposed function for TSPO in mitochondrial cholesterol import, the rate‐limiting step in steroidogenesis. However, this prevalent model was recently refuted based on Tspo gene deletion studies in vivo and in vitro that indicated no deficits in steroidogenesis. Seeking a new functional definition for TSPO, we found that high TSPO expression is not exclusive to steroidogenic cells, but also to other cell types active in lipid metabolism such as liver and adipose tissues. We therefore hypothesized that TSPO plays an important role in mitochondrial fatty acid metabolism. Using steroidogenic MA‐10 Leydig cells that express very high levels of TSPO as the model, we generated CRISPR/Cas9‐mediated TSPO knockout MA‐10 clones (MA‐10:TspoΔ/Δ) and examined changes to core mitochondrial functions after TSPO deletion. MA‐10:TspoΔ/Δ cells showed a shift in substrate utilization for energy production from glucose to fatty acids with significantly higher mitochondrial fatty acid oxidation (FAO), and increased reactive oxygen species (ROS) production. Overexpression of TSPO using a lentiviral system reversely reduced expression of FAO genes in MA‐10 cells. Extending this in vitro observation to in vivo tissues in global TSPO knockout (Tspo−/−) mice, we observed that loss of TSPO in adrenal glands (a major steroidogenic tissue), and liver (the chief metabolic compartment), both significantly upregulated FAO genes compared to the control floxed (Tspofl/fl) tissues. These results demonstrate the first experimental evidence that TSPO affects mitochondrial energy homeostasis through controlling FAO at the level of outer mitochondrial membrane, possibly through regulating activity of carnitine palmitoyltransferase I (CPT1). Therapeutic modulation of TSPO function using available TSPO binding drugs could be beneficial for increasing energy expenditure through FAO in lipid metabolic disorders.Support or Funding InformationStartup funds from Cornell University
Robust Glucose and 1,5‐Anhydroglucitol Uptake Assays Using Novel Fluorescent Conjugates
Authors: Li Shunan , Khatwani Santoshkumar , Lin Yuan , Tchaga Grigority , Yan Gordon 
Published: November 2023

Glucose is the primary carbon source for ATP production and biosynthesis in proliferating mammalian cells by either glycolytic or tricarboxylic acid (TCA) cycle pathways. These pathways are tightly regulated and their disruption leads to many metabolic diseases such as diabetes. Cancer cells exhibit increased glucose uptake to support their high rate of proliferation. The glucose uptake has been widely studied by a non‐radioactive assay using a fluorescently‐labeled glucose derivative, 2‐NBD‐Glucose. Despite its applicability, this conjugate exhibits photo bleaching and reduced fluorescence when used at low concentrations (i.e. <400 μM). BioVision has developed a novel bright green fluorescent 2‐deoxy‐glucose conjugate (GluTracker™) which, just like glucose and 2‐NBD‐Glucose, can be taken up by cells via glucose transporters. However, this glucose analog cannot be fully metabolized by cells, and thus accumulates inside cells. Fluorescence generated by this fluorescent glucose analog is proportional to the cellular glucose uptake and can be used to measure glucose uptake using fluorescence microscopy or flow cytometry. In addition, we have also developed a novel green fluorescent 1,5‐Anhydroglucitol conjugate (AGTrackerTM) to study the uptake of 1,5‐Anhdroglucitol (1,5‐AG). The blood plasma level of 1,5‐AG has been used in clinical studies to monitor short‐term glycemic control in diabetic patients. The AGTrackerTM conjugate can be used to study 1,5‐AG uptake by fluorescence microscopy or flow cytometry. In conclusion, BioVision's GluTracker™ (for Glucose) and AGTrackerTM (for 1,5‐AG) Uptake Assays provide valuable tools for drug discovery and identifying potential modulators in glucose‐dependent pathways.Support or Funding InformationNA
Antidiabetic Effect of the Ethanolic Extracts of Azadirachta indica Leaves on Streptozotocin Induced Wistar Rats
Authors: Aniefuna Ejike Louis , Omorevbarhia Omorovbiye 
Published: November 2023

The antidiabetic efficacy of aqueous extract of Azadirachta indica leaf in normal and streptozotocin induced diabetic rats were compared and evaluated. Phytochemical analyses as well as 12 hrs acute toxicity test of the orally administered ethanolic extract of Azardirachta inidica were carried out using standard methods. Diabetic condition was induced in rats by a single intraperitoneal dose of 200mg/kg b.w. of streptozotocin. The ethanolic extract was administered orally in 2 doses of 50mg/kg b.w. and 100mg/kg for 12 days. The 12 hrs acute toxicity test of the extract of A. indica leaves yield a LD50 of 4.50g/k.g b.w. Both doses of the extract were significantly (p<0.05) effective in reducing blood sugar levels of streptozotocin induced hyperglycaemic rats on the twelveth day of treatment when compared to the control diabetic rats that were treated with gubenclamide (a standard antidiabetic drug) at a dose of 0.2mg/kg b.w on the same day. Nevertheless, 100mg/kg b.w had the highest reduction in blood glucose level. These results suggests that the ethanolic leaf extract of A. indica has antidiabetic effects and could be of benefit in the management of diabetes mellitus as well as in preventing the onset of diabetes mellitus.Support or Funding InformationOmorevbarhia Omorovbiye, graduate student member ASBMB, omonao.omo@gmail.comAniefuna Ejike, graduate student member, ASBMB, aniefunaejike@yahoo.com
Conjugated Linoleic Acids (CLA) and Insulin Resistance: Differential Effects of CLA Isomers on Endoplasmic Reticulum (ER) Stress
Authors: Dammanahalli Jagadeesha K , Subbaiah Papasani V 
Published: November 2023

Conjugated linoleic acids (CLA), the naturally occurring fatty acids prevalent in dairy products, are being used by the general public for potential health benefits, including weight loss, and cancer prevention, but they have also been reported to cause insulin resistance (IR) in animal models. However, the mechanism of action of CLA is not well understood. The current study tested the hypothesis that the development of insulin resistance after consumption of specific CLA isomers is due to the induction of endoplasmic reticulum (ER) stress. We tested the effects of four different CLA isomers, trans‐10,cis‐12 (t10c12)‐, cis 9, trans 11 (c9t11), trans 9 trans 11 (t9t11) and cis 9 cis 11 (c9c11) in three different cell lines‐ HepG2 (hepatocyte), c2c12 (myocyte) and 3T3‐L1 (adipocyte). The cells were treated with 0–100 μM CLA for 24 h, and analyzed for reactive oxygen species (ROS) by flow cytometry, and for ER stress markers by Western blotting. We found that both t9t11, and t10c12‐CLAs induced ROS and the expression of GRP78, the central regulator of ER stress, at 50 μM concentration in HePG2 and 3T3 cells. However these effects were elicited only by the t9t11 isomer in c2c12 cells. GADD153 (CHOP), another marker of ER stress, was induced by t10c12 in HepG2 and 3T3 cells whereas, c9t11, t9t11 and c9c11‐CLAs induced it in 3T3 and c2c12 cells but not in HepG2 cells. Further studies demonstrated that the t10c12 isomer inhibited the insulin‐stimulated phosphorylation of insulin receptor and AKT in HepG2 cells, whereas the c9t11, c9c11 and t9t11 isomers increased the phosphorylation of both proteins. These findings suggest that t10 c12 CLA, the isomer known to cause insulin resistance in mice, may do so by inducing ER stress.Support or Funding InformationSupported by VA Merit Review BX001090
Decreasing O‐GlcNAc Levels Reverses Diabetic Nephropathy‐Associated Phenotypes
Authors: Peterson Sherket , Delannoy Michael , Banerjee Partha , Hart Gerald 
Published: November 2023

Diabetic nephropathy (DN) is a leading cause of morbidity and mortality in diabetic patients. DN is characterized by structural changes of the glomeruli in the kidney, such as mesangial cell expansion, podocyte foot process effacement, and apoptosis, with accompanying glomerular hyperfiltration. Hyperglycemia has been shown to initiate these glomeruli structural changes by stimulating flux through the hexosamine biosynthetic pathway (HBP). The major endpoint of the HBP is the formation of uridine diphosphate β‐D‐N‐acetylglucosamine (UDP‐GlcNAc), the donor for protein O‐GlcNAcylation, and complex extracellular glycosylation. O‐GlcNAcylation is a dynamic nutrient sensitive post‐translational modification that is characterized by the addition of single β‐D‐N‐acetylglucosamine (O‐GlcNAc) to the serine and/or threonine residues of almost every functional class of protein. O‐GlcNAc serves as a nutrient/stress sensor regulating signaling, transcription, cytoskeletal dynamics, and cell division. Altered O‐GlcNAc signaling is directly involved in the pathogenesis of diabetes and may have a role in diabetic complications. However, the molecular mechanisms of how hyperglycemia leads to glomerular dysfunction are largely unknown. Here we report that a dynamic sugar‐protein modification, O‐GlcNAcylation, which serves as a nutrient sensor, links hyperglycemia to defects in glomerular structure and function. By targeting the enzymes that either add or remove O‐GlcNAc (OGT and OGA, respectively), we show that O‐GlcNAcylation regulates mesangial ECM production. Increasing O‐GlcNAc distorts gene expression, protein expression, and subcellular distribution of key slit diaphragm specific proteins, whereas decreasing O‐GlcNAcylation, even in hyperglycemic conditions, restore podocytes to normal. Electron microscopy of the slit diaphragm reveals that decreasing O‐GlcNAc rescues the DN‐associated phenotype. This study provides evidence that O‐GlcNAc plays a role in glomerular structure and function and may contribute to the molecular etiology of DN‐associated phenotypes.Support or Funding InformationThis work is supported by NIH grants F32DK100199 to S. Peterson and R01DK61671, P01HL107153, and N01‐HV‐00240 to G. Hart.
Identification of Novel Antimicrobial Peptides to be used in Functional Foods
Authors: Doering Sean , Thomas Michael , Grant Jennifer E , Lee Eun Joo 
Published: November 2023

Phosvitin, an egg yolk protein similar to casein in milk, is the most phosphorylated protein found in nature. The resulting anionic charge allows phosvitin to bind important bivalent ions such as zinc, iron, magnesium, and calcium. Because of its metal binding properties, phosvitin shows intriguing potential as a source of peptides that could be used in functional foods. Antibacterial properties of phosvitin have come to light as well. In this abstract we report on the antibacterial properties associated with phosvitin, and also discuss work to identify potential peptides that could be used in functional foods as either calcium delivery vehicles or antibacterial agents.When bound to a bivalent cation such as zinc, phosvitin has previously been shown to elicit antimicrobial activity against bacterial species isolated from human hands (unpublished results). Since Serratia marsescens is a hand bacteria associated with human pathology, an isolate of this bacteria was utilized in antibacterial tests of phosvitin activity. Both Kirby Bauer and serial dilution antimicrobial tests were utilized to determine any potential antimicrobial activity. In Kirby Bauer antimicrobial tests, paper discs were soaked in 1mM ZnCl2 solutions with phosvitin in varying concentrations. These discs were then placed on LB plates seeded with S. marsescens and grown at 25 degrees Celsius to determine if rings of inhibition formed around the treated discs. In serial dilution tests, phosvitin was mixed in various concentrations in a 1mM ZnCl2 LB broth solution seeded with S. marsescens. These solutions were then incubated at 25 degrees Celsius to determine if bacterial growth occurred. Minimum inhibitory concentration (MIC) was determined to be around 5 mg/ml phosvitin in 1mM ZnCl2 LB broth solution. These experiments will be repeated using calcium to determine if antimicrobial effects extend across different phosvitin‐bound bivalent cations.Work is also underway to identify potential calcium‐binding peptides derived from phosvitin and identify potential antibacterial activity that may be associated with them. Our peptide of interest is residues 2–13 of phosvitin with a phosphorylation post‐translational modification on residue 12 resulting in the peptide of AEFGTEPDAKpTS. The binding capacity and antimicrobial activity of this peptide will be compared to phosvitin as a potential bioavailable replacement.Support or Funding InformationThis work was supported by the University of Wisconsin Stout Student Research Grant, as well as a Student Jobs Program Award, to S.D.
Genesis of Antibiotic Resistance XII: Trifling Role of SMARXT DISPOSAL TM to Mitigate the Evolution of Antibiotic Resistance
Authors: Kannan Subburaj , Ambriz Paloma , Flores Oscar , Fernandez Priscilla , Carrales Nallely , Mata Eduardo , Moreno Analicia , Herrera Estefania , Hernandez Jazania 
Published: November 2023

SMARXT DISPOSAL has been identified as public‐private partnership between the U.S. Fish and Wildlife Service, the American Pharmacists Association, and the Pharmaceutical Research and Manufacturers of America and North Plains Groundwater Conservation District in Texas) (NPGCD‐Texas). Methodical disposal of unused prescription drugs in NPGCD‐Texas monitored by multiple agencies has been suggested to mitigate the contamination of ground water by the Pharmaceuticals and Personal Care Products (PPCPs) including the antibiotics and antimicrobials. Systematic analysis of the data on file for the pharmaceuticals including the antibiotics and antimicrobials in the water quality data analysis of NPGCD‐Texas has been completed. The analysis of the water quality data show that neither the pharmaceuticals nor the antibiotics/antimicrobials were performed in the Ogallala aquifer. The Ogallala aquifer contains groundwater, with little to no recharge and over 8000 active wells within this district. As identified by the State of Texas, it is a finite and dwindling resource of water in Texas Panhandle. United States Geological Survey (USGS), and Texas Water Development Board(TWDB) presented an antiquated water quality analysis in NPGCD which failed to provide any relevant data on PPCPs in particular antibiotics concentration in the soil and water. Sedimentation analysis on residual metabolite intermediates has not yet been performed. TCEQ's data bases is devoid of either antibiotics or metabolite intermediate or its derivative in NPGCD. Coincidental, lack of water quality analysis for PPCP including antibiotics/antimicrobials is a viable threat to the public health. Data analysis for PPCP, on data communicated by the NPGCD‐Texas will be presented at EB 2016.Support or Funding InformationProfessional development funds provided to S.Kannan at SWTJC for 2014–2015
Genesis of Antibiotic Resistance XIII: Critical Appraisal of incineration of Prescription Drug en masse from National Drug Take Back day (NDTBD) by United States Drug Enforcement Administration – Department of Justice (US‐DEA‐DOJ)
Authors: Kannan Subburaj , Munoz Janet , Luna Jennifer , Rosales Ramiro , Rodriguez Jennifer , Ruiz Vanessa , Velez Micaela , Rodriguez Lizzeth , Chavez Dariela , Subburaj Prabavathy 
Published: November 2023

An effort has been to demonstrate that the incineration of Prescription Drug en masse impart potential environmental hazard. According to http://www.natracare.com/en‐GB/ the current data shows that even in “state‐of‐the‐art” incinerators, burning hazardous waste will lead to the release of three types of dangerous pollutants into our environment: heavy metals; unburned toxic chemicals; and new pollutants ‐ entirely new chemicals formed during the incineration process (Verbatim). According to the US Environmental Protection Authority: “Fugitive emissions and accidental spills may release as much or even more toxic material into the environment than direct emissions from incomplete waste incineration.” According to Greenpeace, studies have identified up to 43 different semi‐volatile, organic chemicals in incinerator ash, and at least 16 organic chemicals in scrubber water from hazardous waste incinerators. Ash is commonly buried in landfill, while effluent is often treated before being discharged into rivers or lakes. Fragments of partially burned waste chemicals recombine within incinerator furnaces, smokestacks, and/or pollution control devices. One study identified 250 volatile organic compounds, many of which are known to be highly toxic or carcinogenic. Among the possible compounds are dioxins and furans, often referred to as just dioxins suppression of the immune system, diabetes, organ toxicity and a wide range of effects on hormones. It is estimated that for every three tonnes of waste that is incinerated, one tonne of ash is generated. Projection analysis on the amount of fly ash generated due to the incineration of drugs from National Drug Take Back day (NDTBD) by United States Drug Enforcement Administration – Department of Justice (US‐DEA‐DOJ) mount 3 fold increase in the environmental contamination. However, taking into account of anaerobic digestion of the drugs from NDTBD – US‐DEA‐DOJ could potentially decrease the methane emissions by 5 – 7 folds compare to the industrial chemicals disposal standard. Taken together, the projection analysis data indicate that there is a statistically significant increase in the environmental contamination by incineration the drugs from NDTBD – US‐DEA‐DOJ. Data analysis for the environmental impact assessment of chemical pollutant will be presented at EB 2016.Support or Funding InformationProfessional Development Funds provided to S. Kannan at SWTJC for 2014–2015