bluuebunny/crossref_metadata_2025_split huggingface dataset:
In Silico Investigation of Gastroprotective Compounds from n‐Butanol Fraction of Costus igneus on Antiulcer Druggable Targets
Authors: Adetayo Modupe O. , Anyasor God'swill N. 
Published: May 2022

Gastric ulcer is a common gastrointestinal tract disorder characterized by discontinuation in the stomach mucosal lining and forming a painful lesion. Costus igneus is an ethnobotanical plant used in the management of gastrointestinal disturbances. This study characterized the gastroprotective bioactive compounds from n‐butanol fraction of C. igneus leaf (CIBF) and investigated their druggability effects on selected antiulcer targets using in silico approach.CIBF compounds were identified and characterized using gas chromatography‐mass spectrometry (GC‐MS) method. The 3D structures of CIBF compounds were downloaded in SDF format from Pubchem National Library of Medicine compound database and used as ligands to dock H+K+ATPase receptor (PDB ID: 5YLV), muscarinic receptor (PDB ID: 5ZHP), urease (PDB ID: 6ZJA) and histamine receptor (using Beta‐1‐adrenergic receptor [PDB ID: 2YCW] as a template). The 3D structures of the proteins were downloaded from the Protein Data Bank. Omeprazole, cimetidine and vonoprazan were used as reference drugs. The ligands and target proteins were processed and prepared for docking in pdb formats by using Chimera 1.14 and OpenBabel GUI version 2.3.2a, respectively. Multi‐targeted molecular docking was done using PyRx – Python Prescription 0.8. Physicochemical and pharmacokinetic properties were analyzed using Osiris DataWarrior version 5.5.0. Bioavailability and drug‐likeness analyses were done using the SwissADME web tool.GC‐MS analysis detected 15 suspected bioactive compounds in CIBF. Molecular docking study showed that 10‐11, (6ʹ,7ʹ) naphthoquinono [3.2] paracyclophane (NQP) had the highest ligand binding affinity for H+K+ATPase, muscarinic receptor, urease and histamine receptor when compared with the others 14 compounds, omeprazole, cimetidine and vonoprazan. Physicochemical properties of NQP did not violate the Lipinski, verber and lead likeness rules. Pharmacokinetic predictive analysis indicated that NQP possesses good oral bioavailability, high gastrointestinal absorption, blood‐brain barrier permeability and no inhibitory effect on cytochrome P450s.Findings from this study showed that NQP exhibited substantial inhibitory effects on proton pump, urease activity, muscarinic and histamine receptors. It also showed good physicochemical and pharmacokinetic properties. It is recommended that n‐butanol fraction of C. igneusleaves should be further explored as a potential source of drug lead candidate for the management of gastric ulcer disease.
Glycosylated Sulfonylurea (2DGs) Modulates Insulin ‐ dependent and Insulin ‐ independent Signaling Pathways via PI3K and P38 MAPK in L6 Skeletal Muscle Cell Line
Authors: Nalweyiso Jacinta , Okechukwu Patrick N. , Sie Ting Ling N. , Hui Tan Y. , Balachandran Abbirami , Siew Ling Crystale L. , Ghadeer Suaifan , Anisa Fromming Gabriele R. , Johnson Stanlas 
Published: May 2022

BackgroundMetformin and sulfonylureas are the most prescribed oral anti‐diabetic drugs globally. However, side effects like lactic acidosis, cardiovascular risks, hypoglycemia and weight gain limit their use in certain comorbidity cases. Metformin utilizes both insulin‐dependent and insulin‐independent sig naling pathways to modulate blood glucose levels in diabetic patients. Sulfonylureas increase insulin secretion from the pancreas and enhance glucose uptake by skeletal muscles through the PI3K signaling pathway. Glycosylated sulfonylurea (2DGs) is a modified analogue of sulfonylurea developed by integrating an aryl sulfonamide with a glucosamine moiety. Previous research on 2DGs has shown in vitro and in vivo antidiabetic, antioxidant, anti‐glycation activity and alleviation from kidney, liver, and cardiovascular diabetic‐related complications.ObjectiveThe current study focused on the evaluation of insulin signaling pathways of 2DGs in normal and insulin resistant L6 myotubes.MethodTo evaluate PI3K signaling, insulin resistance was induced in L6 myotubes using high glucose (25 mmol/l) and insulin (100 nmol/l) for 24 hours, followed by low glucose (5mmol/l) and no serum for 5 hours and insulin (100 nmol/l) for 30 mins. Treatment was done for 24 hours with tolbutamide and acetylated and deacetylated 2DGs at 5uM, 10uM. For p38 MAPK signaling, one group of L6 myotubes was incubated with SB203580 (p38 MAPK inhibitor) and second group was without SB203580. Both groups were treated with 2DGs similar as mentioned above. The treatment was left for 1 hour, after which DNA and protein extraction for quantitative PCR, ELISA, and Western blot experiment was done.ResultsThere was observed high expression of PIK3CA, SLC2A4 and MAPK14 gene in qPCR and upregulated phosphorylation of PI3K, GLUT 4 and p38 MAPK protein in acetylated and deacetylated treated groups compared to tolbutamide and non‐treated group. A positive Pearson correlation was observed between the PIK3CA, SLC2A4 and MAPK14 gene expressions and PI3K, GLUT 4 and p38 MAPK protein expressions respectively.ConclusionFinding indicates that 2DGs insulin signaling pathway seems to be through both insulin dependent via PI3K and insulin ‐ independent via p38 MAPK phosphorylation in insulin resistant and normal L6 myotubes respectively.
Deficient Central Leptin Signaling “In Vivo” Alters Hypothalamic Levels of Circadian Clock Proteins Changing the Diurnal Eating Behavior in Rats
Authors: Mazuecos Lorena , Pintado Cristina , Villar Margarita , Rubio Blanca , Rodriguez Maria , Arribas Carmen , Andres Antonio , Gallardo Nilda 
Published: May 2022

BackgroundLeptin, acting at CNS, decreases food intake (FI), body weight (BW) and adiposity, while circadian dysfunction induces hypothalamic leptin resistance associated to increased adiposity. Hence, we hypothesized that partial inhibition of hypothalamic leptin signaling “in vivo” would alter circadian rhythms resulting in changes in diurnal eating behaviors leading to increased adiposity and peripheral insulin resistance.Methods3‐months‐old Wistar rats were randomly assigned in two different groups: rats infused “in vivo” with saline (PBS) (n=12) or rats infused with a leptin antagonist (SLA) (0.2 micrograms/day) (n=12) via icv minipump insertion for 21 days. On day 0, rats were individually housed in metabolic cages and maintained for 21 days measuring daily BW, FI and locomotor activity. Hypothalamic proteomic analysis (n=6 per group) was carried out to determine if hypothalamic circadian proteins could be altered upon chronic SLA infusion. On day 21 and before sacrifice, one group of rats infused with PBS or SLA (n=6 per group) were injected icv with 100 ng of rat leptin in a volume of 2 microliters. 30 min later rats were euthanatized.ResultsSLA infusion decreased leptin‐induced phosphorylation of pY705‐STAT3 at the hypothalamus sustaining the induction of central leptin resistance upon chronic SLA administration. As a result, daily FI, final BW and visceral adiposity increased in SLA‐infused rats. Remarkedly, the diurnal eating behavior was altered in the SLA group of rats. In fact, daily FI was significantly increased by 48% only during the light phase in SLA infused vs PBS treated control rats, while in the dark phase FI increased less than 10% in the SLA group compared with control PBS‐infused rats. Moreover, SLA‐infused rats develop peripheral insulin resistance assessed by HOMA‐IR at the end of the treatment. Hypothalamic proteomic analysis revealed significant changes in 446 proteins, 266 were up‐regulated and 180 down‐regulated upon SLA infusion. Interestingly, proteins involved in the transcriptional activity of BMAL1/CLOCK were altered in SLA‐infused rats. Paraspeckle component 1, involved in the negative regulation of BMAL1/CLOCK, was over‐represented, while vasopressin‐neurophysin 2 copeptin, a positive regulator of circadian gene expression, was under‐represented.ConclusionOur results suggest that deficient hypothalamic leptin signaling is associated to circadian dysfunction which leads to changes in the diurnal eating behavior in rats resulting in increased adiposity and peripheral insulin resistance.
A Connection Between DNA Repair Protein APE1, Alpha‐Synucleinopathy, and Biological Sex in Rodents and Humans
Authors: Miner Kristin M. , Bhatia Tarun N. , Jamenis Anuj S. , Clark Rachel N. , Abraham Nevil , Rajasundaram Dhivyaa , Sauvaigo Sylvie , Stetler R. A. , Chen Jun , Sanders Laurie H. , Luk Kelvin C. , Leak Rehana K. 
Published: May 2022

Lewy body disorders are characterized by proteostatic and redox disequilibrium, leading to deposition of α‐synuclein in hallmark inclusions and oxidative damage to DNA. One common pathway for repair of oxidative DNA damage is base excision repair (BER), which involves the coordinated activity of several enzymes, including apurinic/apyrimidinic endonuclease 1 (APE1). The main goal of this study was to assess APE1 in rodents and humans with α‐synucleinopathy, and to test the functional impact of APE1 in the preformed α‐synuclein fibril model. First, we report that knockdown of APE1 with two independent shRNA sequences or inhibition of APE1 DNA repair activity increased inclusions bearing pathologically‐phosphorylated α‐synuclein (pSer129) in preformed fibril‐treated primary hippocampal cultures. Second, we examined APE1 expression in a mouse model of limbic‐centered α‐synucleinopathy, in which α‐synuclein fibrils are infused into the olfactory bulb/anterior olfactory nucleus (OB/AON). Six months later, we observed a fibril‐induced decrease in APE1 expression in the brains of male mice and an increase in females.Similar sex‐opposing patterns were noted for APE1 mRNA expression. Third, we demonstrated that the loss of APE1 in fibril‐infused male mice in vivo is mediated by oxidative stress, as APE1 loss was abolished by dietary administration of the antioxidant N‐acetylcysteine. Fourth, OB/AON tissues harvested from fibril‐infused male mice displayed higher cleavage of synthetic DNA lesions on a biochip than tissues collected in parallel from fibril‐infused female mice. However, OB/AON tissues harvested from fibril‐infused female mice had higher DNA repair activity compared to males. Assessments of open field activity measures and sucrose preference revealed a negative behavioral impact of α‐synuclein fibril infusions in male but not female mice. Fifth, men with Lewy body disorders displayed lower APE1 expression in the OB and amygdala compared to women with Lewy body disorders, similar to fibril‐infused mice. Finally, we infused α‐synuclein fibrils into the OB/AON of transgenic APE1 overexpressing rats and observed a reduction in pSer129 levels in male and female brains compared to fibril‐infused wildtype rats. Preliminary data suggest that APE1 overexpression in fibril‐infused rats improved olfaction and anxiety measures, especially in males. These findings reveal a sex‐biased impact of limbic α‐synucleinopathy on APE1 expression and function but also suggest that APE1 overexpression attenuates α‐synucleinopathy in both sexes.
Monocytes transition to monocyte‐macrophages within the inflamed vasculature via CCR2 on monocytes and endothelial TNFR2
Authors: Sathyanarayana Mysore Vijayashree , Tahir Suhail , Furuhashi Kazuhiro , Arora Jatin , Rosetti Florencia , Cullere Xavier , Yazbeck Pascal , Sekulic Miroslav , Lemieux Madeleine E. , Raychaudhuri Soumya , Horwitz Bruce , Mayadas Tanya 
Published: May 2022

ObjectiveTo understand the cellular pathways that drive the maturation of classical monocytes into macrophages in inflammationHypothesisMonocytes play essential roles in homeostasis, infection and sterile inflammation and can acquire macrophage phenotypes by transitioning through intermediate states. The mechanisms leading to the accumulation of these monocyte intermediates within tissues is not well understood; in the intestine, it has been attributed to a disruption in the differentiation along the monocyte–macrophage continuum under steady state and following inflammation. We hypothesize that monocytes double positive for chemokine receptors, CX3CR1 and CCR2 are preferentially recruited to the glomerulus and transition to immature, inflammatory macrophages while still within the vasculature in a CCR2 dependent manner.MethodsMonocyte recruitment and maturation was evaluated using mouse models of acute/ chronic glomerulonephritis, intravital microscopy of the kidney, cell sorting, multi‐parameter flow cytometry and adoptive transfer approaches. Mechanistic insights into this conversion were provided using in vivo intravascular staining, in vitro monocyte‐endothelial cell co‐cultures and single cell transcriptional profiling. Evidence of the conversion was also evaluated by flow cytometric analysis of blood and urine samples from SLE patients with active nephritis.ResultsWe show that renal Ly6C+ monocytes acquire macrophage markers (MHCII, CD64, F4/80) upon renal inflammation and are distinguished by the expression of both CCR2 and CX3CR1. These cells, which we refer to as immature macrophages, may be derived from a CCR2+CX3CR1+ double‐positive monocytes in blood that are preferentially recruited and retained within glomerular capillaries, and acquire pro‐inflammatory characteristics. Mechanistically, the monocyte acquisition of macrophage markers occurs within the vasculature and relies on CCR2 on hematopoietic lineages and TNFR2 on parenchymal cells, findings that are recapitulated in vitro with monocytes co‐cultured with TNF‐TNFR2 activated endothelial cells, which generate CCR2 ligands. Single‐cell RNA‐sequencing defines a monocyte differentiation path associated with the acquisition of immune effector functions, and monocyte CCR2‐dependent generation of CCR2 ligands. Immature macrophages are detected in the urine of patients with lupus nephritis and their frequency correlates with clinical disease.ConclusionsCCR2 dependent functional specialization of monocytes into immature macrophages occurs within the TNF‐TNFR2 activated vasculature and establishes a chemokine‐based autocrine, feed‐forward loop that may amplify inflammation and renal injury.
Lipidomics identifies novel circulating markers of CVD risk in African American and Caucasian women
Authors: Gonzalez Paula , Jain Raghav , Coe Chris , Ryff Carol , King Camille , Bersh Andy , Malecki Kristen , Simcox Judith 
Published: May 2022

Cardiovascular disease (CVD) is the leading cause of mortality for women in the USA. Women with dyslipidemia have at increased risk of CVD. The parameters of dyslipidemia have primarily been defined using Caucasian male populations. Current lipid markers of dyslipidemia such as high density lipoprotein (HDL), low density lipoprotein (LDL), and triglycerides have been shown to be poor predictors of CVD risk in African American populations. We aim to identify marks of dyslipidemia in African American populations that better predict risk and development of CVD.Untargeted lipidomics (total lipid #) was performed on sera from middle‐age African American and Caucasian women (n=872). Machine learning was used to identify predictive lipid markers of CVD risk. Findings were validated using an independent cohort of women (n=200).We identified unique and shared signatures of CVD in women based on race. Arachidonic acid containing lipids are positively correlated with elevated blood pressure and cardiomyopathy. Because these lipids are important in inflammatory signaling, we also assessed interleukin‐6 and c‐reactive protein. Through machine learning we were able to determine that markers of inflammation and inflammatory lipids were predictive of CVD in both Caucasian and African American women. Next, we will perform targeted lipidomics on these lipids of interest.Regardless of class, lipid species containing arachidonic acid were increased in circulation of women with hypertension. We propose the consideration of an inflammatory marker panel that includes arachidonic acid to assess CVD risk in women.
The Effects of Exercise Training with Oral Intake of Lactate on Hippocampal Neurogenesis and Neurotrophic Factors in Mice
Authors: Hwang Deunsol , Kyun Sunghwan , Jang Inkwon , Park Hun‐Young , Kim Jisu , Lim Kiwon 
Published: May 2022

Hippocampal neurogenesis plays an important role in learning and memory. Healthy hippocampal environment is also necessary for improvement of its cognitive function. Therefore, enhancing of both hippocampal neurogenesis and environment is required to prevent or ameliorate neurodegenerative statements.Exercise is powerful enhancer of both, and it is considered that the exercise effects are resulted from alterations in brain (central mechanism), also in soma, i.e., skeletal muscle, (somatic mechanism). Yet the exact mechanism is unclear.Lactate, known as just byproduct of glycolysis and primary factor of fatigue when exercise, is highlighting as a promising signaling molecule that mediates the exercise effects on hippocampus. In this perspective, exercise training with intake of lactate can be remarkable strategy for strengthening the exercise effects on hippocampus.Therefore, the purpose of the current study is to investigate whether combination of exercise and lactate intake can additively enhance exercise‐induced hippocampal neurogenesis and neurotrophic factors.The study was conducted for 5 weeks using 4 groups of mice: CON (vehicle group), LAC (oral administration of 3g/kg lactate 5days/week), EXE (mid‐intensity exercise 5days/week), and EXE+LAC (lactate administration immediately after every exercise). At the last week of experiment, cognitive function was evaluated by Y‐maze spontaneous alternation test. After the experiment ended, hippocampal neurogenesis (Ki67, neural stem cell proliferation marker; DCX, marker of differentiation to immature neurons) and neurotrophic factors (PGC1α, FNDC5, VEGFA, and BDNF) were evaluated respectively by immunohistochemistry (IHC) and by Western blot (WB) on hippocampus.From the IHC results, exercise training significantly increased the number of Ki67 and DCX positive cells (p= 0.001 and p = 0.004 respectively) which were also increased in LAC than CON (p = 0.064 and p = 0.03 respectively). There was no difference in Ki67 and DCX positive cells between EXE and EXE+LAC. From the WB results, however, FNDC5 of EXE+LAC was significantly higher than that of EXE (p = 0.005). BDNF and PGC1α of EXE+LAC was higher than that of EXE respectively by 20% and 9% (but not significant). There was no difference in VEGFA between EXE and EXE+LAC. From the Y‐maze test results, interestingly, while there was no difference in the performance between CON and LAC, the performance of EXE+LAC was significantly enhanced than that of EXE (p = 0.006).In conclusion, combination of exercise training and lactate intake additively promoted the performance of spontaneous alternation test and tended to additively enhance exercise‐induced neurotrophic factors, but not hippocampal neurogenesis. Therefore, the study revealed the potential that exercise training with oral intake of lactate can be remarkable strategy for strengthening the exercise effects on hippocampus.
Role of apoptosis and autophagy in masseter muscle atrophy evoked by Botulinum Toxin injection
Authors: Quezada Esteban R. , Chiong Mario , Buvinic Sonja 
Published: May 2022

IntroductionMusculoskeletal disorders of the masticatory system impact the quality of life and have a high cost of diagnosis and treatment. Masseter muscle hyperactivity is a cause of pain in the chewing apparatus. Botulinum toxin type A (BoNTA) injection is widely used to induce paralysis of the masseter muscle, thereby decreasing impaired muscle activity. However, our laboratory has described in a preclinical model that the injection of BoNTA not only induces paralysis, but also muscle atrophy, which subsequently decreases bone quality. However, it is unknown whether apoptosis or autophagy mechanisms could contribute to muscle atrophy.AimTo evaluate the induction of apoptosis and autophagy in BoNTA‐injected masseter muscle of adult mice.MethodologyUnilateral injection of BoNTA (0.2U/10µl) in the masseter muscle was performed in adult BalbC mice (approved by IACUC‐Universidad de Chile, #21446‐ODO‐UCH). Apoptosis and autophagy markers were evaluated in masseter muscles by immunoblot at 2‐7d post‐injection and immunofluorescence at 7d post‐injection. Autophagy activity was blocked by i.p injection of chloroquine. The data were evaluated with t‐test, one‐way ANOVA test, Mann‐Whitney test, or Kruskal‐Wallis test, as appropriate. The results were expressed as mean ±SEM (n=4‐8; p <0.05).ResultsUnilateral injection of BoNTA did not change the relative levels of apoptosis‐related proteins like cleaved Caspase 3, PARP, and AIF. There was a significant increase in protein levels related to autophagy, such as lipidated LC3 (1.86‐fold), non‐lipidated LC3 (1.66‐fold), and P62 (1.24‐fold) at 7 d post‐injection. Also, there was a strong punctuated stain for LC3 in histological sections of masseter muscle 7d post injection, suggesting autophagic vesicles. However, the injection of chloroquine for blocking the autophagy flux did not improve the accumulation of autophagy proteins in masseter muscle evoked by BoNTA in the induction.ConclusionsThe atrophy of masseter muscle evoked by BoNTA injection is not related to autophagy‐ or apoptosis‐induction. The increase in autophagy markers after BoNTA injection, with no further increase after autophagic flux blockade, suggests that BoNTA may be blocking autophagy in the masseter muscle, perhaps favoring a proteasomal pathway of muscle atrophy.